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  • photosynthesis  (93)
  • Immunohistochemistry  (88)
  • Springer  (181)
  • American Institute of Physics (AIP)
  • Cambridge University Press
  • Periodicals Archive Online (PAO)
  • 2020-2023
  • 2020-2022
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  • 1935-1939
  • 1994  (110)
  • 1990  (71)
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  • 1990-1994  (181)
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  • 1
    ISSN: 1572-8773
    Keywords: Chorella vulgaris ; acid tolerance ; ATPase ; nickel toxicity ; nutrient uptake ; photosynthesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract This study concerns the inhibitory effects of acid pH and nickel on growth, nutrient (NO3 - and NH4 +) uptake, carbon fixation, O2 evolution, electron transport chain and enzyme (nitrate reductase and ATPase) activities of acid tolerant and wild-type strains of Chlorella vulgaris. Though a general reduction in all these variables was noticed with decreasing pH, the tolerant strain was found to be metabolically more active than the wild-type. A reduced cation (NH4 +, Na+, K+ and Ca2+) uptake, coupled with a facilitated influx of anions (NH4 +, PO4 3- and HCO3 -), suggested the development of a positive membrane potential in acid tolerant Chlorella. Nevertheless, a tremendous increase in ATPase activity at decreasing pH revealed the involvement of superactive ATPase in exporting H+ ions and keeping the internal pH neutral. A difference in Na+ and K+ efflux of the two strains at decreasing pH suggests there is a difference in membrane permeability. The low toxicity of Ni in the acid tolerant strain may be due to the low Ni uptake brought about by a change in membrane potential as well as in permeability. Hence, the development of superactive ATPase and a change in both membrane potential and permeability not only offers protection against acidity, but also co-tolerance to metals.
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  • 2
    ISSN: 1432-0827
    Keywords: Decorin ; Proteoglycan-100 ; Heterotopic ossification ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Heterotopic ossification is a metabolically active process which shares several properties of orthotopic bone formation and, therefore, represents an excellent model for studying bone matrix components. Immunohistochemical methods were used to investigate the distribution pattern of the small proteoglycans decorin and proteoglycan-100 during different stages of heterotopic ossification of pressure sores of paraplegic patients. Decorin and proteoglycan-100 exhibited a substantially divergent distribution pattern. Decorin was detectable in the perivascular matrix of granulation tissue as well as in the stroma of heterotopic ossification. The ossification zone was stained most strongly. In contrast, proteoglycan-100 was predominantly detectable in fibroblasts and preosteoblasts in early areas of osteogenesis. In more mature forms of heterotopic ossification immunostaining was markedly reduced in osteoblasts and osteocytes and even absent in so-called bone-lining cells. However, at least some osteoclasts were strongly positive. These results suggest indicate that decorin and proteoglycan-100 are important components during the formal pathogenesis of heterotopic ossification. The expression of the small proteoglycans, especially of proteoglycan-100, correlates with different phases during heterotopic ossification, showing a maximum for proteoglycan-100 in matrix-forming cells in early phases of bone formation, but in osteoclasts in mature bone.
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Calcified tissue international 55 (1994), S. 38-45 
    ISSN: 1432-0827
    Keywords: Vacuolar-type H+-ATPase ; Carbonic anhydrase II ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract The localization of vacuolar-type H+-ATPase and carbonic anhydrase II (CA II) in rat incisor enamel organs at maturation was examined by light and electron microscopy. The immunoreactivity for both vacuolar-type H+-ATPase and CA II was intense on the ruffled border of ruffle-ended ameloblasts (RA), but moderate at the distal end of smooth-ended ameloblasts (SA). Immuno-gold particles indicated that CA II was not confined to the ruffled border of RA alone, but also distributed in the cytoplasm of RA and SA. These findings suggest that RA may secrete protons produced by CA II via the ruffled border into enamel by active transport of vacuolar-type H+-ATPase. Secreted protons may activate hydrolytic enzymes to degrade the organic components of enamel matrix. Vacuolar-type H+-ATPase on vesicles of SA suggests that a specific configuration of ruffled borders in RA may be formed by the fusion of vesicle membranes in the distal end of cytoplasm of SA.
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  • 4
    ISSN: 1573-5028
    Keywords: carbonic anhydrase ; antisense ; over-expression ; tobacco ; photosynthesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The activity and location of carbonic anhydrase has been modified by transformation of tobacco with antisense and over-expression constructs. Antisense expression resulted in the inhibition of up to 99% of carbonic anhydrase activity but had no significant impact on net CO2 assimilation. Stomatal conductance and susceptibility to water stress appeared to increase in response to the decline in carbonic anhydrase activity. An over-expression construct designed to increase cytosolic carbonic anhydrase abundance resulted in a significant increase in net activity, a small increase in stomatal conductance but little impact on CO2 assimilation. Chloroplastic carbonic anhydrase activity was enhanced by the expression of an additional construct which targeted the polypeptide to the organelle. The increase in chloroplastic carbonic anhydrase appeared to be accompanied by a concomitant increase in Rubisco activity.
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  • 5
    ISSN: 1573-5028
    Keywords: gene expression ; photosynthesis ; protein turnover ; psbA ; tac promoter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The unicellular cyanobacterium Synechococcus sp. PCC 7942 has three psbA genes encoding two different forms of the photosystem II reaction centre protein D1 (D1:1 and D1:2). The level of expression of these psbA genes and the synthesis of D1:1 and D1:2 are strongly regulated under varying light conditions. In order to better understand the regulatory mechanisms underlying these processes, we have constructed a strain of Synechococcus sp. PCC 7942 capable of over-producing psbA mRNA and D1 protein. In this study, we describe the over-expression of D1:1 using a tac-hybrid promoter in front of the psbAI gene in combination with lacI Q repressor system. Over-production of D1:1 was induced by growing cells for 12 h at 50 μmol photons m-2 s-1 in the presence of 40 or 80 μg/ml IPTG. The amount of psbAI mRNA and that of D1:1 protein in cells grown with IPTG was three times and two times higher, respectively. A higher concentration of IPTG (i.e., 150 μg/ml) did not further increase the production of the psbAI message or D1:1. The over-production of D1:1 caused a decrease in the level of D1:2 synthesised, resulting in most PSII reaction centres containing D1:1. However, the over-production of D1:1 had no effect on the pigment composition (chlorophyll a or phycocyanin/number of cells) or the light-saturated rate of photosynthesis. This and the fact that the total amounts of D1 and D2 proteins were not affected by IPTG suggest that the number of PSII centres within the membranes remained unchanged. From these results, we conclude that expression of psbAI can be regulated by using the tac promoter and lacI Q system. However, the accumulation of D1:1 protein into the membrane is regulated by the number of PSII centres.
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  • 6
    ISSN: 1432-0878
    Keywords: Key words: NADPH diaphorase ; Nitric oxide synthase ; Nervous system ; central ; Nervous system ; peripheral ; Immunohistochemistry ; Helix aspersa (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Enzyme histochemistry and immunocytochemistry were used to determine the distribution of neurons in the snail Helix aspersa which exhibited nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase activity and/or immunoreactivity to nitric oxide synthase (NOS). NADPH diaphorase-positive cells and fibres were distributed extensively throughout the central and peripheral nervous system. NADPH diaphorase-positive fibres were present in all neuropil regions of the central and peripheral ganglia, in the major interganglionic connectives and in peripheral nerve roots. NADPH diaphorase-positive cell bodies were found consistently in the eyes, the lips, the tentacular ganglia and the procerebral lobes of the cerebral ganglia; staining of cell bodies elsewhere in the nervous system was capricious. The distribution of NOS-like immunoreactivity differed markedly from that of NADPH diaphorase activity. Small clusters of cells which exhibited NOS-like immunoreactivity were present in the cerebral and pedal ganglia; fibres which exhibited NOS-like immunoreactivity were present in restricted regions of the neuropil of the central ganglia. The disjunct distributions of NADPH diaphorase activity and NOS-like immunoreactivity in the nervous system of Helix suggest that the properties of neuronal NOS in molluscs may differ sigificantly from those described previously for vertebrate animals.
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 277 (1994), S. 457-464 
    ISSN: 1432-0878
    Keywords: Mammary-derived growth inhibitor ; Fatty acid-binding proteins ; Differentiation ; Vascularization ; Immunohistochemistry ; Immunocytochemistry ; Mammary gland ; Cow
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Mammary-derived growth inhibitor (MDGI) has previously been localized in the mammary parencyma, dependent on the stage of differentiation of the mammary gland. Here, we have elucidated the distribution of MDGI in the mammary stroma by a combined immunohisto-and cytochemical analysis with antibodies raised against MDGI. Distinct staining of capillary endothelial cells has been revealed. Although its subcellular distribution resembles former observations in secretory epithelial cells, the expression of MDGI in capillary endothelial cells clearly precedes that in secretory epithelial cells. On the other hand, no endothelial MDGI staining has been detected in bovine heart, which contains a fatty acid-binding protein almost identical to MDGI. The localization of MDGI in the mammary capillary endothelium is discussed in terms of its possible involvement in the intracellular transport of hydrophobic ligands or in the regulation of endothelial cell proliferation.
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  • 8
    ISSN: 1432-0878
    Keywords: Key words: Chromogranins ; Serotonin ; Amylin ; Regulatory peptides ; Gut ; Monoclonal antibodies ; Immunohistochemistry ; Rana esculenta (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Novel monoclonal antibodies to human chromogranin A (CgA) and chromogranin B (CgB) were used to investigate the presence of immunoreactive (-IR) elements in the alimentary tract of the green frog Rana esculenta. Numerous CgA-IR and a few CgB-IR endocrine cells were found within the gut mucosa, from the oesophagus to the cloaca, with some local differences in density. Co-localization studies demonstrated that they were co-stored in almost all the serotonin-IR, the amylin-IR or islet amyloid polypeptide-IR cells and in the peptide tyrosine tyrosine-IR cells located proximal to the pylorus, but not in those located in more caudal tracts. No other co-localization was demonstrated; substances investigated included somatostatin, substance P, gastrin/cholecystokinin, glucagon, glycentin, bombesin, secretin and neurotensin. CgA-IR and CgB-IR cells nearly always displayed argyrophilia with the Grimelius silver method
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  • 9
    ISSN: 1432-0878
    Keywords: Chromogranins ; Serotonin ; Amylin ; Regulatory peptides ; Gut ; Monoclonal antibodies ; Immunohistochemistry ; Rana esculenta (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Novel monoclonal antibodies to human chromogranin A (CgA) and chromogranin B (CgB) were used to investigate the presence of immunoreactive (-IR) elements in the alimentary tract of the green frog Rana esculenta. Numerous CgA-IR and a few CgB-IR endocrine cells were found within the gut mucosa, from the oesophagus to the cloaca, with some local differences in density. Co-localization studies demonstrated that they were costored in almost all the serotonin-IR, the amylin-IR or islet amyloid polypeptide-IR cells and in the peptide tyrosine tyrosine-IR cells located proximal to the pylorus, but not in those located in more caudal tracts. No other co-localization was demonstrated; substances investigated included somatostatin, substance P, gastrin/cholecystokin, glucagon, glycentin, bombesin, secretin and neurotensin. CgA-IR and CgB-IR cells nearly always displayed argyrophilia with the Grimelius silver method
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  • 10
    ISSN: 1432-0878
    Keywords: Key words: Corpuscles of Stannius ; Stanniocalcin ; Immunocytochemistry ; Immunohistochemistry ; Western blot ; Lepisosteus osseus (Holostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Stanniocalcin-immunoreactive cells were localized in the corpuscles of Stannius of a holostean fish, the garpike (Lepisosteus osseus), using antisera against salmon and trout stanniocalcins and the peroxidase-antiperoxidase and protein A-gold immunohistochemical methods. The stanniocalcin-immunoreactive cells were periodic acid-Schiff-positive, and antibody staining was abolished if the antiserum was preabsorbed with corpuscle homogenate. Immunocytochemistry revealed two reactive cell types in the glandular parenchyma, and immunoreactivity was confined to the secretory granules. Staining of the granules was also abolished when the antisera were blocked with crude corpuscle homogenate. When corpuscle extracts from garpike were subjected to sodium dodecyl sulphate-polyacrylamide gel electrophoresis and Western blot analysis, a single dense band was evident with a molecular weight of ∼68 kDa under non-reducing conditions, whereas three bands were observed (∼29, ∼31, and ∼34 kDa) under reducing conditions. Staining of all bands disappeared following preabsorption of the antiserum with salmon stanniocalcin, trout stanniocalcin, or garpike corpuscle extract. The results are compared with stanniocalcins from another extant holostean, the bowfin (Amia calva), and from more modern bony fishes, the teleosts.
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  • 11
    ISSN: 1432-0878
    Keywords: Corpuscles of Stannius ; Stanniocalcin ; Immunocytochemistry ; Immunohistochemistry ; Western blot ; Lepisosteus osseus (Holostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Stanniocalcin-immunoreactive cells were localized in the corpuscles of Stannius of a holostean fish, the garpike (Lepisosteus osseus), using antisera against salmon and trout stanniocalcins and the peroxidase-antiperoxidase and protein A-gold immunohistochemical methods. The stanniocalcin-immunoreactive cells were periodic acid-Schiff-positive, and antibody staining was abolished if the antiserum was preabsorbed with corpuscle homogenate. Immunocytochemistry revealed two reactive cell types in the glandular parenchyma, and immunoreactivity was confined to the secretory granules. Staining of the granules was also abolished when the antisera were blocked with crude corpuscle homogenate. When corpuscle extracts from garpike were subjected to sodium dodecyl sulphate-polyacrylamide gel electrophoresis and Western blot analysis, a single dense band was evident with a molecular weight of ∼68 kDa under non-reducing conditions, whereas three bands were observed (∼29, ∼31, and ∼34 kDa) under reducing conditions. Staining of all bands disappeared following preabsorption of the antiserum with salmon stanniocalcin, trout stanniocalcin, or garpike corpuscle extract. The results are compared with stanniocalcins from another extant holostean, the bowfin (Amia calva), and from more modern bony fishes, the teleosts.
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  • 12
    Electronic Resource
    Electronic Resource
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    Cell & tissue research 277 (1994), S. 325-331 
    ISSN: 1432-0878
    Keywords: c-Myc ; c-Fos ; Vasoactive intestinal polypeptide (VIP) ; Immunohistochemistry ; Intestine, small ; Enteric nervous system ; Guinea pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Immunohistochemical double labelling of the enteric nervous system of the guinea pig ileum was performed with a monoclonal antibody (anti-MYC 033) directed against a peptide sequence of the human c-Myc protein together with antibodies directed against either the neuron-specific antigens neuron-specific enolase or PGP 9.5 or the glia-specific marker S-100 to demonstrate that anti-MYC 033 labelled the nuclei of all enteric neurons but not glia. This strategy was also employed to demonstrate that another anti-c-Myc monoclonal anti-body, anti-MYC 070, labelled the nuclei of all neurons and glia, as well as perhaps all other cells in these preparations. A polyclonal antiserum raised against a peptide sequence of the human c-Fos protein (anti-FOS 4) was shown to label the identical nuclei as anti-MYC 033. The ganglionic density of nuclei labelled by anti-FOS 4 was found to be similar to previous measures of the ganglionic density of neurons. Double labelling with anti-MYC 033 and an antiserum directed against vasoactive intestinal polypeptide was performed to reexamine the ganglionic density of neurons that express this neuropeptide. Our results suggest that the ganglionic density of these neurons might be less than previously determined.
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  • 13
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 277 (1994), S. 325-331 
    ISSN: 1432-0878
    Keywords: Key words: c-Myc ; c-Fos ; Vasoactive intestinal polypeptide (VIP) ; Immunohistochemistry ; Intestine ; small ; Enteric nervous system ; Guinea pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Immunohistochemical double labelling of the enteric nervous system of the guinea pig ileum was performed with a monoclonal antibody (anti-MYC 033) directed against a peptide sequence of the human c-Myc protein together with antibodies directed against either the neuron-specific antigens neuron-specific enolase or PGP 9.5 or the glia-specific marker S-100 to demonstrate that anti-MYC 033 labelled the nuclei of all enteric neurons but not glia. This strategy was also employed to demonstrate that another anti-c-Myc monoclonal antibody, anti-MYC 070, labelled the nuclei of all neurons and glia, as well as perhaps all other cells in these preparations. A polyclonal antiserum raised against a peptide sequence of the human c-Fos protein (anti-FOS 4) was shown to label the identical nuclei as anti-MYC 033. The ganglionic density of nuclei labelled by anti-FOS 4 was found to be similar to previous measures of the ganglionic density of neurons. Double labelling with anti-MYC 033 and an antiserum directed against vasoactive intestinal polypeptide was performed to reexamine the ganglionic density of neurons that express this neuropeptide. Our results suggest that the ganglionic density of these neurons might be less than previously determined.
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  • 14
    ISSN: 1432-0878
    Keywords: Central nervous system ; Neuropeptides ; Immunohistochemistry ; Amphioxus, Branchiostoma belcheri (Cephalochorda)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The immunohistochemical localization of nine different neuropeptides was studied in the central nervous system of the amphioxus, Branchiostoma belcheri. In the brain, perikarya immunoreactive for urotensin I and FMRFamide were localized in the vicinity of the central canal. One of the processes of each of these perikarya was found to cross the dorso ventral slit-like lumen of the central canal. Oxytocin-immunoreactive short fibers, but not perikarya, were detected in the ventral part of the brain. Perikarya immunoreactive for arginine vasopressin/vasotocin, oxytocin and FMRFamide were widely distributed in the spinal cord. Arginine vasopressin/vasotocin-immunoreactive fibers often made contacts with Rohde cell axons. Angiotensin II-immunoreactive perikarya were observed in the posterior half of the spinal cord, and urotensin I-immunoreactive perikarya were found in the caudal region of the spinal cord. Cholecystokinin/gastrin-immunoreactive fibers, but not perikarya, were detected in the spinal cord; some extended as far as the ependymal layer of the cerebral ventricle. No colocalization of the peptides examined was observed. No immunoreactivity for atrial and brain natriuretic peptides nor for urotensin II was detected. The present study indicates that there are at least six separate neuronal systems that contain different peptides, respectively, in the central nervous system of the amphioxus. Their functions remain to be determined.
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  • 15
    ISSN: 1432-0878
    Keywords: NADPH diaphorase ; Immunohistochemistry ; Gastrointestinal tract ; Nitric oxide ; Histochemistry ; Guinea-pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The distribution and abundance of nitric oxide synthase (NOS)-containing neurons and their terminals in the gastrointestinal tract of the guinea-pig were examined in detail using NADPH diaphorase histochemistry and NOS immunohistochemistry. NOS-containing cell bodies were found in the myenteric plexus throughout the gastrointestinal tract and in the submucous plexus of the stomach, colon and rectum. NOS-containing neurons comprised between 12% (in the duodenum) and 54% (in the esophagus) of total myenteric neurons. In the ileum, NOS neurons represented 19% of total myenteric neurons. Most of the NOS neurons throughout the gastrointestinal tract possessed lamellar dendrites and a single axon. NOS-containing terminals were abundant in the circular muscle, including that of the sphincters, but were rare in the longitudinal muscle, except for the taeniae of the caecum. The muscularis mucosae of the esophagus, stomach, colon and rectum received a medium to dense innervation by NOS terminals. Within myenteric ganglia, NOS-containing terminals were extremely sparse in the esophagus, stomach and duodenum, common in the ileum and distal colon and extremely dense in the proximal colon and rectum. The submucous plexus in the ileum and large intestine contained a sparse plexus of NOS-containing terminals. NOS terminals were not observed in the mucosa of any region. We conclude that throughout the gastrointestinal tract of the guinea-pig, NOS neurons are inhibitory motor neurons to the circular muscle; in the ileum and large intestine, NOS neurons may also function as interneurons.
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  • 16
    ISSN: 1432-0878
    Keywords: Substance P ; Calcitonin gene ; related peptide ; Cervical spinal nerve ; Immunohistochemistry ; Primary afferents ; Mouse (ICR)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We demonstrate the existence of nerve fibers possessing substance P (SP) and calcitonin gene-related peptide (CGRP) immunoreactivity in the mouse cervical ventral roots. The distribution of the SP and CGRP fibers was similar, but CGRP fibers were generally more numerous. Both types entered the ventral pia mater or formed hairpin loops, but they did not enter the spinal cord directly through these roots. SP and CGRP fibers in the ventral roots were thin and had many varicosities. We suggest that these SP and CGRP fibers are involved not only in a sensory mechanism, but also in other functions, via the release of SP and CGRP from varicosities in the ventral roots.
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  • 17
    ISSN: 1432-0878
    Keywords: Key words: Pituitary ; Gonadotrophs ; LHβ ; FSHβ ; Immunohistochemistry ; Rana japonica (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Immunohistochemical localization of lutropin β (LHβ) and follitropin β (FSHβ) in the pituitary gland of the frog Rana japonica was studied by the peroxidase-anti-peroxidase method and the two-face, double-labeling method with different-sized gold particles at the light- and electron-microscopic levels, respectively, using monoclonal antibodies against bullfrog LHβ and FSHβ. Light-microscopic immunohistochemistry indicated that approximately 66.0% of all the gonadotrophs in the pituitary contained both LHβ and FSHβ, whereas 33.4% of gonadotrophs contained only LHβ, and 0.6% contained only FSHβ. The staining intensity of LHβ and FSHβ varied from cell to cell. The gonadotrophs were classified into four types (Types I-IV) in terms of their ultrastructural and immunolabeling characteristics. Moreover, several secretory granule types were recognized according to differences in their shape and electron density. In all the cell types, both LHβ and FSHβ were often seen in the same secretory granules, but the proportion of granules bearing both hormones ranged from 5.5% in Type I to 32.7% in Type IV. Most secretory granules in Types I and II were immunolabeled with LHβ alone, whereas a small number of granules were immunolabeled with FSHβ alone. More immunolabeled FSHβ granules were present in Types III and IV than in Types I and II.
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  • 18
    ISSN: 1432-0878
    Keywords: NADPH diaphorase ; Nitric oxide synthase ; Nervous system, central ; Nervous system, peripheral ; Immunohistochemistry ; Helix aspersa (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Enzyme histochemistry and immunocytochemistry were used to determine the distribution of neurons in the snail Helix aspersa which exhibited nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase activity and/or immunoreactivity to nitric oxide synthase (NOS). NADPH diaphorase-positive cells and fibres were distributed extensively throughout the central and peripheral nervous system. NADPH diaphorase-positive fibres were present in all neuropil regions of the central and peripheral ganglia, in the major interganglionic connectives and in peripheral nerve roots. NADPH diaphorase-positive cell bodies were found consistently in the eyes, the lips, the tentacular ganglia and the procerebral lobes of the cerebral ganglia; staining of cell bodies elsewhere in the nervous system was capricious. The distribution of NOS-like immunoreactivity differed markedly from that of NADPH diaphorase activity. Small clusters of cells which exhibited NOS-like immunoreactivity were present in the cerebral and pedal ganglia; fibres which exhibited NOS-like immunoreactivity were present in restricted regions of the neuropil of the central ganglia. The disjunct distributions of NADPH diaphorase activity and NOS-like immunoreactivity in the neurvous system of Helix suggest that the properties of neuronal NOS in molluscs may differ sigificantly from those described previously for vertebrate animals.
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  • 19
    ISSN: 1432-0878
    Keywords: Pituitary ; Gonadotrophs ; LHβ ; FSHβ ; Immunohistochemistry ; Rana japonica (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Immunohistochemical localization of lutropin β (LHβ) and follitropin β (FSHβ) in the pituitary gland of the frog Rana japonica was studied by the peroxidase-anti-peroxidase method and the two-face, double-labeling method with different-sized gold particles at the light-and electron-microscopic levels, respectively, using monoclonal antibodies against bullfrog LHβ and FSHβ. Light-microscopic immunohistochemistry indicated that approximately 66.0% of all the gonadotrophs in the pituitary contained both LHβ and FSHβ, whereas 33.4% of gonadotrophs contained only LHβ, and 0.6% contained only FSHβ. The staining intensity of LHβ and FSHβ varied from cell to cell. The gonadotrophs were classified into four types (Types I–IV) in terms of their ultrastructural and immunolabeling characteristics. Moreover, several secretory granule types were recognized according to differences in their shape and electron density. In all the cell types, both LHβ and FSHβ were often seen in the same secretory granules, but the proportion of granules bearing both hormones ranged from 5.5% in Type I to 32.7% in Type IV. Most secretory granules in Types I and II were immunolabeled with LHβ alone, whereas a small number of granules were immunolabeled with FSHβ alone. More immunolabeled FSHβ granules were present in Types III and IV than in Types I and II.
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  • 20
    Electronic Resource
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    Cell & tissue research 277 (1994), S. 457-464 
    ISSN: 1432-0878
    Keywords: Key words: Mammary-derived growth inhibitor ; Fatty acid-binding proteins ; Differentiation ; Vascularization ; Immunohistochemistry ; Immunocytochemistry ; Mammary gland ; Cow
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Mammary-derived growth inhibitor (MDGI) has previously been localized in the mammary parenchyma, dependent on the stage of differentiation of the mammary gland. Here, we have elucidated the distribution of MDGI in the mammary stroma by a combined immunohisto- and cytochemical analysis with antibodies raised against MDGI. Distinct staining of capillary endothelial cells has been revealed. Although its subcellular distribution resembles former observations in secretory epithelial cells, the expression of MDGI in capillary endothelial cells clearly precedes that in secretory epithelial cells. On the other hand, no endothelial MDGI staining has been detected in bovine heart, which contains a fatty acid-binding protein almost identical to MDGI. The localization of MDGI in the mammary capillary endothelium is discussed in terms of its possible involvement in the intracellular transport of hydrophobic ligands or in the regulation of endothelial cell proliferation.
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  • 21
    ISSN: 1432-0878
    Keywords: FMRFamide ; ACEP-1 (Achatina cardio-excitatory peptide-1) ; Cardiac regulation ; Immunohistochemistry ; Achatina fulica (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Immunohistochemical localization of two neuropeptides possibly involved in the regulation of cardiac activity in a pulmonate mollusc, Achatina fulica Férussac, was studied. On the ventral surface of the right cerebral ganglion, more than 50 neurons with diameters of 30–50 μm showed immunoreactivity to the antiserum of the neuropeptide FMRFamide. Many were also immunoreactive to an antiserum raised against Achatina cardio-excitatory peptide-1 (ACEP-1). Although FMRFamidelike immunoreactive neurons occurred in all components of the subesophageal ganglia, identifiable ACEP-1-like immunoreactive neurons were located only in the visceral ganglion and the right parietal ganglion. In the heart, FMRFamide- and ACEP-1-like immunoreactive fibers were restricted to the atrium and the aortic end of the ventricle, consistent with morphological observations of cardiac innervation. The present results suggest that FMRFamide-and ACEP-1-like peptides are involved in regulating the heart beat of this snail.
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  • 22
    ISSN: 1432-0878
    Keywords: Neurotoxins ; Immunohistochemistry ; Macrophages ; Dendritic reticulum cell ; B-cells ; Indoleamines ; NADPH oxidase ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Antibodies to quinolinic acid were produced in rabbits with protein-conjugated and gold particle-adsorbed quinolinic acid. Quinolinic acid immunoreactivity was below detection limits in carbodiimide-fixed rat brain. In contrast, strong quinolinic acid immunoreactivity was observed in spleen cells with variable, complex morphology located predominantly in the periarterial lymphocyte sheaths. In the thymus, quinolinic acid immunoreactivity was observed in cells with variable morphology, located almost exclusively in the medulla. Lymph nodes and gut-associated lymphoid tissue contained many, strongly stained cells of similar complex morphology in perifollicular areas. Immunoreactivity in liver and lung was restricted to widely scattered, perivascular cells and alveolar cells respectively. Additional stained cells with complex morphology were observed in bronchus-associated lymphoid tissue, in skin, and in the lamina propria of intestinal villi. Follicles in all secondary lymphoid organs were diffusely stained, ranging from mildly to moderately immunoreactive in spleen, to intensely immunoreactive in gut-associated lymphoid tissue. These results suggest that quinolinic acid is an immune system-specific molecule. Two hypothetical schemes are proposed to account for high levels of quinolinic acid in specific cells of the immune system.
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  • 23
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    Cell & tissue research 278 (1994), S. 573-578 
    ISSN: 1432-0878
    Keywords: Key words: Placenta ; Giant cells ; Chorionic gonadotropin ; Luteotropin ; Electrophoresis ; Immunohistochemistry ; Rabbit
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Obplacental giant cells are enlarged cells, found following implantation, in the antimesometrial region of the rabbit uterus. They probably originate from trophoblastic knobs that traverse the uterine epithelium during early implantation. Little is known about their function. In this study, trophoblast, placental, paraplacental and obplacental tissues at days 7–15 post-coitum, and enzyme-isolated giant cells at day 15 were studied by two-dimensional gel electrophoresis, followed by immunoblotting and light-microscopic immunohistochemistry, for the presence of human chorionic gonado- tropin-like proteins. Immunostaining was performed by using anti-human chorionic gonadotropin antibodies. In gel electrophoresis of obplacental tissue and isolated giant cells, two proteins of human chorionic gonadotropin-like antigenicity at 26 kDa with pIs equivalent to pH 6.4 and 6.6 were found; they were absent in the placenta, paraplacenta, day-7 blastocyst and day-8 trophoblast. The onset of synthesis of these proteins could be observed when day-8 trophoblastic tissue was cultured in vitro for 24 h. In immunohistochemistry, only the obplacental giant cells showed a positive reaction, indicating that the production of chorionic gonadotropin occurs in this cell type.
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  • 24
    ISSN: 1432-0878
    Keywords: Key words: FSH ; Immunohistochemistry ; Receptor mRNA ; In situ hybridization ; Sertoli cell ; Testis ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Testicular biopsies from 82 oligo- or azoospermic male patients were subjected to immunostaining using anti-human FSH antibodies. Histological evaluation showed normal spermatogenesis (nspg) in 7 (FSH: 2.7±0.7), mixed atrophy (ma) in 63 (FSH: 5.3±0.5), and bilateral or unilateral Sertoli Cell Only syndrome (SCO) in 12 (FSH:21.7±3.5) patients. For the relationship between FSH values and testicular histology, see Bergmann et al. (1994). FSH immunoreactivity was found exclusively in Sertoli cells and in some interstitial cells. Seminiferous epithelium showing normal or impaired spermatogenesis displayed only weak immunoreactivity compared to intense immunoreaction, i.e. large and numerous vesicles in Sertoli cells of SCO tubules in biopsies showing mixed atrophy or SCO. In addition, h-FSH receptor mRNA was demonstrated by in situ hybridization using biotinylated cDNA antisense oligonucleotides. Hybridization signals were found within the seminiferous epithelium exclusively in Sertoli cell cytoplasm associated with normal spermatogenesis and in epithelia showing different signs of impairment, including SCO. It is concluded that: (1) Sertoli cells are the only cells within the seminiferous epithelium expressing FSH receptors; (2) the accumulation of FSH immunoreactivity in Sertoli cells of SCO tubules appears to be a sign of impaired Sertoli cell function.
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  • 25
    ISSN: 1432-0878
    Keywords: Carotid labyrinth ; Coexistence ; Substance P ; CGRP ; VIP ; Neuropeptide Y ; Immunohistochemistry ; Rana catesbeiana (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Double immunohistochemical staining with rhodamine- and fluorescein isothiocyanate (FITC)-conjugated antisera revealed the coexistence of substance P (SP) and neuropeptide Y (NPY), and SP and calcitonin gene-related peptide (CGRP) in most nerve fibers in the intervascular stroma of the carotid labyrinth of the bull-frog, Rana catesbeiana, although there were a few fibers which showed only SP- or NPY-immunoreactivity. Approximately one third of SP-immunoreactive fibers also showed coexistence with vasoactive intestinal polypeptide (VIP)-immunoreactivity, and a few fibers contained VIP without SP. The combination of the double immunofluorescence technique and alternate consecutive sections further demonstrated the possible coexistence of SP, VIP, NPY, and CGRP. This coexistence of four different peptides in the same nerve fibers was proved by the following two evident facts: 1) some SP fibers which demonstrated coexistence with NPY-immunoreactivity were assumed to be continuous with those showing VIP-immunoreactivity, and 2) almost all of the SP fibers showed coexistence with CGRP-immunoreactivity. By this reasoning, nearly one third of SP fibers may demonstrate coexistence with NPY-, VIP-, and CGRP-immunoreactivities. These multiple peptides might be involved in vascular regulatory function, which is a possible function of the amphibian carotid labyrinth.
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  • 26
    ISSN: 1432-0878
    Keywords: Key words: Pineal organ ; Neuron-specific enolase ; Immunohistochemistry ; Three-dimensional reconstruction ; Post-hatching development ; Domestic fowl
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Immunohistochemistry for neuron-specific enolase (NSE) revealed that NSE is localized in both a limited number of pinealocytes and intrinsic afferent neurons in the pineal organ of the domestic fowl. Furthermore, a computer-assisted three-dimensional imaging technique allowed to clarify the reverse distributional pattern of both elements: NSE-positive pinealocytes displayed a dense distribution especially in the vesicular portion of the gland, whereas NSE-immunoreactive nerve cells were mainly found in the pineal stalk. The number of NSE-positive intrinsic neurons in the pineal organ of chickens decreased rapidly after hatching, with a concentration of these elements in the basal portion (stalk) of the pineal organ. On the other hand, immunoreactive pinealocytes increased remarkably in the end-vesicle of the organ with age, followed by a gradual expansion toward the proximal portion. Thus, the spectacular increase in NSE-positive pinealocytes and the progressive reduction of reactive neurons occurred in parallel during the course of post-hatching development. NSE-immunoreactive pinealocytes displayed morphological characteristics of bipolar elements, endowed with an apical protrusion into the pineal lumen and a short basal process at younger stages, whereas multipolar types of NSE-positive pinealocytes were predominantly found in the adult domestic fowl. These results indicate that in the pineal organ of the domestic fowl (1) the ontogenetic expansion of NSE-immunoreactive pinealocytes is paralleled by a regressive afferent innervation, (2) the NSE-positive pinealocytes transform from a bipolar (columnar) type to a multipolar type during post-hatching development, and (3) these ontogenetic changes in the NSE-immunoreactivity and morphology of pinealocytes may reflect the development of a neurosecretory-like capacity of the organ.
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  • 27
    ISSN: 1432-0878
    Keywords: Key words: Pharynx ; Lung ; Calcitonin gene-related peptide ; Substance P ; Coexistence ; Immunohistochemistry ; Rana catesbeiana (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Indirect double immunofluorescence labelling in the pharynx and lung of the bullfrog, Rana catesbeiana, demonstrated the occurrence, distribution, and coexistence of two neuropeptides. In the pharynx, immunoreactive calcitonin gene-related peptide (CGRP) and substance P (SP) were localized in nerve fibers distributed within and just beneath the ciliated epithelium. In the lung, CGRP and SP were localized in nerve fibers in five principal locations: 1) within the smooth muscle layer in the interfaveolar septa; 2) in the luminal thickened edges of the septa; 3) around the pulmonary vasculature; 4) within, and 5) under the ciliated epithelium. Within the smooth muscle layer in the septa, luminal thickened septa, and around blood vessels, almost all fibers showed coexistence of CGRP and SP. Within and just beneath the ciliated epithelium in the thickened septa, all fibers showed coexistence of CGRP and SP. No immunoreactivity for vasoactive intestinal polypeptide, neuropeptide Y, galanin, somatostatin, FMRFamide, and leucine- and methionine-enkephalins was detected in the nerve fibers within the larynx and the lung. Together with our previous data, the present findings suggest that peptidergic mechanisms are involved in the regulation of amphibian respiratory systems throughout their life.
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  • 28
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    Cell & tissue research 279 (1994), S. 169-181 
    ISSN: 1432-0878
    Keywords: Key words: Adrenal gland ; Dorsal root ganglia ; Immunohistochemistry ; Neurofilament ; Neuronal tracing ; Neuropeptides ; Nitric oxide synthase ; Substance P ; Guinea-pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Retrograde neuronal tracing in combination with double-labelling immunofluorescence was applied to distinguish the chemical coding of guinea-pig primary sensory neurons projecting to the adrenal medulla and cortex. Seven subpopulations of retrogradely traced neurons were identified in thoracic spinal ganglia T1-L1. Five subpopulations contained immunolabelling either for calcitonin gene-related peptide (CGRP) alone (I), or for CGRP, together with substance P (II), substance P/dynorphin (III), substance P/cholecystokinin (IV), and substance P/nitric oxide synthase (V), respectively. Two additional subpopulations of retrogradely traced neurons were distinct from these groups: neurofilament-immunoreactive neurons (VI), and cell bodies that were nonreactive to either of the antisera applied (VII). Nerve fibres in the adrenal medulla and cortex were equipped with the mediator combinations I, II, IV and VI. An additional meshwork of fibres solely labelled for nitric oxide synthase was visible in the medulla. Medullary as well as cortical fibres along endocrine tissue apparently lacked the chemical code V, while in the external cortex some fibres exhibited code III. Some intramedullary neuronal cell bodies revealed immunostaining for nitric oxide synthase, CGRP or substance P, providing an additional intrinsic adrenal innervation. Perikarya, immunolabelled for nitric oxide synthase, however, were too few to match with the large number of intramedullary nitric oxide synthase-immunoreactive fibres. A non-sensory participation is also supposed for the particularly dense intramedullary network of solely neurofilament-immunoreactive nerve fibres. The findings give evidence for a differential sensory innervation of the guinea-pig adrenal cortex and medulla. Specific sensory neuron subpopulations suggest that nervous control of adrenal functions is more complex than hitherto believed.
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  • 29
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    Cell & tissue research 279 (1994), S. 209-213 
    ISSN: 1432-0878
    Keywords: Key words: Calcium-binding proteins ; Immunohistochemistry ; Mesonephros ; Metanephros ; Chick embryo (White leghorn)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The ontogeny of two calcium-binding proteins (calbindin-D28k and calretinin) was studied by immunohistochemical techniques in developing chick kidney. This study showed the presence of calbindin on the 5th incubation day and calretinin on the 7th incubation day in mesonephric distal and connecting tubules, and in the medial wall of the Wolffian duct. At later stages, immunostaining for these two proteins, in particular for calretinin, was also demonstrated in some metanephric proximal tubules. Glomeruli and Bowman's capsules were negative both in the mesonephros and metanephros. The presence of calretinin in the developing kidney has thus been demonstrated for the first time. The early expression of calbindin and calretinin in mesonephric distal tubules suggests their role in regulating the final excretion of calcium. The different patterns of immunoreactivity of the walls of the Wolffian duct can be correlated with their different histogenetic and histological features.
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  • 30
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    Cell & tissue research 276 (1994), S. 213-221 
    ISSN: 1432-0878
    Keywords: Gut-associated lymphoid tissue (GALT) ; M(membranous)-cells ; Immunohistochemistry ; Cytokeratins ; Yeast ; Pig (Minipig, Göttingen)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The intermediate filaments of the dome epithelium of porcine Peyer's patches were studied by immunohistochemistry. The labelling patterns of monospecific antibodies directed against cytokeratins 8, 18 and 19 differed considerably. About 40% of the dome epithelial cells were intensely labelled by three different anti-cytokeratin 18 antibodies, indicating that large amounts of cytokeratin 18 are present in these cells. In order to verify that these cytokeratin-18-immunoreactive cells were M-cells, uptake studies using fluorescein-labelled yeast particles were performed. Numerous yeast particles were found exclusively in dome epithelial cells that were highly positive for cytokeratin 18, thus representing M-cells. In contrast, the content of cytokeratin 19 in M-cells was lower than that in neighbouring enterocytes. The labelling intensity of cytokeratin 8 did not differ between M-cells and enterocytes. In addition, the absence of vimentin and desmin from the dome epithelium of porcine Peyer's patches was demonstrated. The results show (1) that porcine M-cells differ from enterocytes in the composition of their cytoskeleton, (2) that cytokeratin 18 is a useful marker for detecting porcine M-cells and (3) that this marker directly correlas with M-cell function.
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  • 31
    ISSN: 1432-0878
    Keywords: Subcommissural organ ; Reissner's fiber ; Secretory products ; Immunohistochemistry ; Development, phylogenetic ; Class-specific epitopes ; Dogfish, Scyliorhinus canicula (Elasmobranchii)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We have raised antisera against extracts of the subcommissural organ (SCO) of the dogfish, Scyliorhinus canicula L. Brains of 2900 specimens were collected in acetone, and the region containing the SCO and posterior commissure was removed and extracted in three different media. Antisera against these crude extracts were raised in rats and rabbits. Sequential absorptions of the antisera with extracts from different regions of the dogfish brain were performed to eliminate unwanted antibodies. When used to immunostain sections of the whole central nervous system of the dogfish, these purified antisera reacted selectively with the SCO-Reissner's fiber complex. An antiserum against bovine Reissner's fiber was also used. The antisera against the dogfish SCO and bovine Reissner's fiber showed the same staining pattern in the SCO and the Reissner's fiber of the dogfish. For comparative purposes, the brains of 15 vertebrate species from all vertebrate classes were immunostained with both antisera. The anti-dogfish SCO serum reacted with the SCO of the dogfish and that of other phylogenetically related elasmobranch species. Neither the SCO of a primitive elasmobranch species, Heptranchias perlo, nor the SCO of the other classes of vertebrates reacted with the anti-dogfish SCO serum. However, the antiserum against bovine Reissner's fiber reacted with the SCO of all the investigated species. It is concluded that some epitopes (or compounds) in the secretory material of the SCO are class-specific, whereas others are conserved and are synthesized by the SCO in most vertebrate species.
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  • 32
    ISSN: 1432-0878
    Keywords: Key words: FMRFamide ; ACEP-1 (Achatina cardio-excitatory peptide-1) ; Cardiac regulation ; Immunohistochemistry ; Achatina fulica (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Immunohistochemical localization of two neuropeptides possibly involved in the regulation of cardiac activity in a pulmonate mollusc, Achatina fulica Férussac, was studied. On the ventral surface of the right cerebral ganglion, more than 50 neurons with diameters of 30–50 μm showed immunoreactivity to the antiserum of the neuropeptide FMRFamide. Many were also immunoreactive to an antiserum raised against Achatina cardio-excitatory peptide-1 (ACEP-1). Although FMRFamide-like immunoreactive neurons occurred in all components of the subesophageal ganglia, identifiable ACEP-1-like immunoreactive neurons were located only in the visceral ganglion and the right parietal ganglion. In the heart, FMRFamide- and ACEP-1-like immunoreactive fibers were restricted to the atrium and the aortic end of the ventricle, consistent with morphological observations of cardiac innervation. The present results suggest that FMRFamide- and ACEP-1-like peptides are involved in regulating the heart beat of this snail.
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  • 33
    ISSN: 1432-0878
    Keywords: Key words: Neurotoxins ; Immunohistochemistry ; Macrophages ; Dendritic reticulum cell ; B-cells ; Indoleamines ; NADPH oxidase ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Antibodies to quinolinic acid were produced in rabbits with protein-conjugated and gold particle-adsorbed quinolinic acid. Quinolinic acid immunoreactivity was below detection limits in carbodiimide-fixed rat brain. In contrast, strong quinolinic acid immunoreactivity was observed in spleen cells with variable, complex morphology located predominantly in the periarterial lymphocyte sheaths. In the thymus, quinolinic acid immunoreactivity was observed in cells with variable morphology, located almost exclusively in the medulla. Lymph nodes and gut-associated lymphoid tissue contained many, strongly stained cells of similar complex morphology in perifollicular areas. Immunoreactivity in liver and lung was restricted to widely scattered, perivascular cells and alveolar cells respectively. Additional stained cells with complex morphology were observed in bronchus-associated lymphoid tissue, in skin, and in the lamina propria of intestinal villi. Follicles in all secondary lymphoid organs were diffusely stained, ranging from mildly to moderately immunoreactive in spleen, to intensely immunoreactive in gut-associated lymphoid tissue. These results suggest that quinolinic acid is an immune system-specific molecule. Two hypothetical schemes are proposed to account for high levels of quinolinic acid in specific cells of the immune system.
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  • 34
    ISSN: 1432-0878
    Keywords: Adrenal gland ; Nitric oxide synthase ; Nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH) ; Immunohistochemistry ; Histochemistry ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The distribution and colocalization of nitric oxide synthase and nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-diaphorase) was investigated in the adrenal gland of developing, adult and aging rats with the use of immunohistochemical and histochemical techniques. Nitric oxide synthase-immunoreactive neurons within the adrenal gland were found from the 20th day of gestation onwards. During early development the neurons were found as small clusters of smaller-size cells compared to those observed in the adult gland. Their number reached that of adult level by the 4th day after birth, and in the glands from aging rats a 28.6% increase was observed. Whilst no immunofluorescence was seen in chromaffin cells during early development, some cells from glands of aging rats showed nitric oxide synthase-immunoreactivity with varying intensity. The immunoreactive neurons from postnatal rat adrenals were also positive for NADPH-diaphorase, whilst those in prenatal rats were negative or lightly stained. Nitric oxide synthase-immunoreactive nerve fibres were present in all adrenal glands examined from the 16th day of gestation onwards. A considerable degree of variation in the distribution of immunoreactive fibres both in medulla and outer region of cortex at the different age groups was observed and described. Most, but not all, nitric oxide synthase-immunoreactive nerve fibres also showed NADPH-diaphorase staining.
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  • 35
    ISSN: 1432-0878
    Keywords: Eye ; Lens ; Development, ontogenetic ; αA-crystallin ; αB-crystallin ; Immunohistochemistry ; Human ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The distribution of αA- and αB-crystallin in the developing lens of human (Carnegie stages 13 to 23) and rat embryos (embryonic days E11 to 18) was examined immunohistochemically. In a human embryo at stage 13, the lens placode was already immunoreactive to αB-crystallin, but not to αA-crystallin. At stage 15, the lens vesicle was intensely immunoreactive both to αA- and αB-crystallin. From stages 16 to 23, the lens epithelial cells and fiber cells were immunoreactive to αA- and αB-crystallin. In rat embryos, αA-crystallin appeared in the lens pit at E12, and αB-crystallin appeared in the elongating lens fiber cells at E14. From E15 to E18, the lens epithelial cells and fiber cells were immunoreactive to αA-crystallin. The lens fiber cells were also immunoreactive to αB-crystallin, but the epithelial cells were not. These findings suggest that αB-crystallin appears earlier than αA-crystallin in the human lens, but at a later period than αA-crystallin in the rat lens. αB-Crystallin was not detected in the epithelial cells of the rat lens, but was perisistently present in the epithelial cells of the human lens.
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  • 36
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    Cell & tissue research 276 (1994), S. 381-386 
    ISSN: 1432-0878
    Keywords: Gizzard ; Development, ontogenetic ; Muscle smooth ; Capillaries ; Immunohistochemistry ; Myosin ; Domestic fowl
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Antibodies to smooth muscle and non-muscle myosin allow the development of smooth muscle and its capillary system in the embryonic chicken gizzard to be followed by immunofluorescent techniques. Although smooth muscle development proceeds in a serosal to luminal direction, angiogenetic cell clusters develop independently at the luminal side close to the epithelial layer, and the presumptive capillaries invade the developing muscle in a luminal to serosal direction. The smooth muscle and non-muscle myosin heavy chains in this avian system cannot be separated by SDS polyacrylamide gel electrophoresis and do not show isoform specificity in immunoblotting, unlike the system found in mammals. Only two myosin heavy chains with Mr of 200 and 196 kDa were separable and considerable immunological cross-reactivity was found between the denatured myosin isoform heavy chains.
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  • 37
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    Cell & tissue research 276 (1994), S. 403-410 
    ISSN: 1432-0878
    Keywords: Sympathetic nervous system ; Enteric nervous system ; Noradrenaline ; Catecholamine histofluorescence ; Immunohistochemistry ; Pteropus poliocephalus, P. scapulatus (Chiroptera)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The distribution of catecholamines in the small and large intestine of flying foxes (Pteropus spp.) was investigated using glyoxylic-acid-induced fluorescence and immunohistochemical staining of tyrosine hydroxylase and dopamine-β-hydroxylase. Dense networks of varicose axons stained by each of these methods supplied blood vessels, the mucosa and both submucous and myenteric ganglia, but were scarce in the circular and longitudinal muscle. The majority (〉90%) of submucous neuronal perikarya contained both enzymes and most of these also exhibited catecholamine fluorescence. Somata of similar staining characteristics were less common in the myenteric plexus, where single cells were found in only the minority of ganglia. All of the stained submucosal somata and mucosal axons contained vasoactive intestinal peptide, whereas catecholamine-containing axons that supplied the ganglia, external muscle and blood vessels did not. It is concluded that (1) there is dense catecholamine innervation of most tissues in the flyingfox intestine, similar to many other mammals, (2) mucosal axons originate from enteric catecholamine neurons, not found in other mammals, and (3) axons supplying the blood vessels and enteric ganglia are probably of sympathetic origin and can be distinguished from the intrinsic catecholamine-containing axons by their lack of vasoactive intestinal peptide. The roles and interactions of these two types of catecholamine innervation in the control of secretion and motility remain to be identified.
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  • 38
    ISSN: 1432-0878
    Keywords: Glucagon-like peptide 1 ; Endocrine tumors ; Immunohistochemistry ; Ultrastructure ; Multiple endocrine neoplasia type 1 ; Co-localization ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The preproglucagon gene encodes, in addition to glucagon, two smaller peptides with structural similarity: glucagon-like peptides 1 and 2. Glucagon-like peptide 1 (GLP-1) 7–36 amide is the most powerful incretin candidate. In the present study, GLP-1 immunoreactivity was investigated in tissue specimens of various types of gastroenteropancreatic tumors, and the serum-levels of GLP-1 were assayed. Immunohistochemical staining of 88 tumors revealed GLP-1 immunoreactivity in 17 neoplasias (19.3 %), viz., in 7 out of 33 non-functioning tumors, 4 out of 20 gastrinomas, 4 out of 13 insulinomas, 1 out of 3 vasoactive-intestinal-polypeptide (VIP)omas and 1 adrenocorticotropic-hormone (ACTH)-producing tumor. In these tumors, GLP-1-immunoreactive cells were distributed either diffusely, arranged in clusters, or as single cells. All GLP-1-positive tumors were immunoreactive for glucagon or glicentin, 10 tumors were immunoreactive for pancreatic polypeptide, and 8 tumors for insulin. Ultrastructural analysis of 8 GLP-1-positive tumors, with the immunogold technique, demonstrated GLP-1 immunoreactivity mainly in cells resembling the A-cells of the pancreas or the L-cells of the gut. Of the 17 GLP-1-immunoreactive tumors, 15 were primarily located in the pancreas. Additionally, 2 non-functioning tumors of the rectum were GLP-1 immunoreactive. Five tumors were GLP-1 immunoreactive from 9 patients with multiple endocrine neoplasia I syndrome. Patients with GLP-1-immunoreactive tumors were characterized by a significantly lower rate of distant metastases (P〈0.01) and a higher rate of curative resections (P〈0.05). In 2 out of 22 patients, elevated serum-levels of GLP-1 were found: one patient with a vasoactive-intestinal-polypeptide (VIP)oma and 1 patient with a non-functioning tumor. This indicates that GLP-1 might be secreted at least by a few gastroenteropancreatic endocrine tumors.
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  • 39
    ISSN: 1432-0878
    Keywords: Suprachiasmatic nucleus ; Retinohypothalamic tract ; Immunohistochemistry ; Neuropeptides ; Cytochrome oxidase ; Acetylcholinesterase ; Circadian system ; Domestic sheep
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The retinal innervation, cytoarchitectural, and immunohistochemical organization of the suprachiasmatic nucleus (SCN) was studied in the domestic sheep. The SCN is a large elongated nucleus extending rostrocaudally for roughly 3 mm in the hypothalamus. The morphology is unusual in that the rostral part of the nucleus extends out of the main mass of the hypothalamus onto the dorsal aspect of the optic chiasm. Following intraocular injection of wheat-germ agglutininhorseradish peroxidase or tritiated amino acids, anterograde label is distributed throughout the SCN. Retinal innervation of the SCN is bilaterally symmetric or predominantly ipsilateral. Quantitative image analysis demonstrates that, although the amount of autoradiographic label is greatest in the ventral and central parts of the nucleus, density varies progressively between different regions. In addition to the SCN, retinal fibers are also seen in the medial preoptic area, the anterior and lateral hypothalamic areas, the dorsomedial hypothalamus, the retrochiasmatic area, and the basal telencephalon. Whereas the SCN can be identified using several techniques, complete delineation of the nucleus requires combined tract tracing, cytoarchitectural, and histochemical criteria. Compared with the surrounding hypothalamic regions, the SCN contains smaller, more densely packed neurons, and is largely devoid of myelinated fibers. Cell soma sizes are smaller in the ventral SCN than in the dorsal or lateral parts, but an obvious regional transition is lacking. Using Nissl, myelin, acetylcholinesterase, and cytochrome oxidase staining, the SCN can be clearly distinguished in the rostral and medial regions, but is less differentiated toward the caudal pole. Immunohistochemical demonstration of several neuropeptides shows that the neurochemical organization of the sheep SCN is heterogeneous, but that it lacks a distinct compartmental organization. Populations of different neuropeptide-containing cells are found throughout the nucleus, although perikarya positive for vasoactive intestinal polypeptide and fibers labeled for methionine-enkephalin are predominant ventrally; neurophysine-immunoreactive cells are more prominent in the dorsal region and toward the caudal pole. The results suggest that the intrinsic organization of the sheep SCN is characterized by gradual regional transitions between different zones.
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  • 40
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    Cell & tissue research 278 (1994), S. 171-175 
    ISSN: 1432-0878
    Keywords: Key words: Tachykinin ; Substance P ; Sinus venosus ; Heart ; Immunohistochemistry ; Dogfish ; Scyliorhinus canicula (Elasmobranchii)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The sinus venosus of the elasmobranch heart is characterized by the presence of large bundles of unmyelinated nerve fibres that bulge into the cardiac lumen, below the endocardium. In the dogfish (Scyliorhinus canicula), these fibres contain numerous dense-core membrane-bounded granules of about 200 nm in diameter. Most intramural ganglion cells of the sinus venosus also show densely packed granules similar to those found in the subendocardial fibres. We have observed strong substance-P-like immunoreactivity in the large fibre bundles and in the perikarya of the ganglion cells. Preabsorption of the antisera with fragment 7–11 of substance P has shown that the antisera recognize the tachykinin canonic sequence. Our findings suggest that an undetermined tachykinin is secreted in the elasmobranch heart, and that it is probably released into the blood stream in the context of a little-known neuroendocrine system.
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  • 41
    ISSN: 1432-0878
    Keywords: Key words: Carotid body ; Chief cells ; Catecholamine ; Serotonin ; γ-Aminobutyric acid ; Immunohistochemistry ; Mouse (BALB/c)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The immunohistochemical study revealed tyrosine hydroxylase (TH), dopamine β-hydroxylase (DBH), phenylethanolamine N-methyltransferase (PNMT), serotonin, glutamate decarboxylase (GAD) and γ-aminobutyric acid (GABA) immunoreactivities in the mouse carotid body. TH and DBH immunoreactivities were found in almost all chief cells and a few ganglion cells, and in relatively numerous varicose nerve fibers of the carotid body. The histofluorescence microscopy showed catecholamine fluorescence in almost all chief cells. However, no PNMT immunoreactivity was observed in the carotid body. Serotonin, GAD and GABA immunoreactivities were also seen in almost all chief cells of the carotid body. From combined immunohistochemistry and fluorescence histochemistry, catecholamine and serotonin or catecholamine and GABA were colocalized in almost all chief cells. Thus, these findings suggest that noradrenaline, serotonin and GABA may be synthesized and co-exist in almost all chief cells of the mouse carotid body and may play roles in chemoreceptive functions.
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  • 42
    ISSN: 1432-0878
    Keywords: Key words: EGF ; Cartilage ; Growth plate ; Hypophysectomy ; Growth hormone ; Immunohistochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Epiphyseal growth plate cartilages from the proximal tibia of normal, hypophysectomized, and growth hormone (GH)-treated hypophysectomized rats were subjected to immunohistochemistry for detection of epidermal growth factor (EGF). In the normal growth plate, EGF was distributed mainly in the proliferative zone. Hypophysectomy resulted in considerable atrophy of the chondrocytes and the cartilage matrix (a decreased number of mature-type chondrocytes and a decreased ratio of proliferating to hypertrophic chondrocytes) and a significant diminution of EGF immunoreactivity. Treatment with GH reversed these effects of hypophysectomy, causing an increased thickness of the growth plate and EGF-reactive sites in all chondrocyte layers. The most intense immunostaining for EGF, however, was frequently seen in the nuclei of chondrocytes with flattened appearance. It appears that EGF could be incorporated or synthesized in chondrocytes having marked mitogenic activity. The present results, taken with previous data on EGF involvement in growth of cartilaginous tissue in vivo and in vitro, strongly suggest that EGF-immunoreactive chondrocytes are involved in cartilage proliferation and growth under the specific influence of GH.
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  • 43
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    Cell & tissue research 278 (1994), S. 573-578 
    ISSN: 1432-0878
    Keywords: Placenta ; Giant cells ; Chorionic gonadotropin ; Luteotropin ; Electrophoresis ; Immunohistochemistry ; Rabbit
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Obplacental giant cells are enlarged cells, found following implantation, in the antimesometrial region of the rabbit uterus. They probably originate from trophoblastic knobs that traverse the uterine epithelium during early implantation. Little is known about their function. In this study, trophoblast, placental, paraplacental and obplacental tissues at days 7–15 post-coitum, and enzyme-isolated giant cells at day 15 were studied by two-dimensional gel electrophoresis, followed by immunoblotting and light-microscopic immunohistochemistry, for the presence of human chorionic gonadotropin-like proteins. Immunostaining was performed by using anti-human chorionic gonadotropin antibodies. In gel electrophoresis of obplacental tissue and isolated giant cells, two proteins of human chorionic gonadotropin-like antigenicity at 26 kDa with pIs equivalent to pH 6.4 and 6.6 were found; they were absent in the placenta, paraplacenta, day-7 blastocyst and day-8 trophoblast. The onset of synthesis of these proteins could be observed when day-8 trophoblastic tissue was cultured in vitro for 24 h. In immunohistochemistry, only the obplacental giant cells showed a positive reaction, indicating that the production of chorionic gonadotropin occurs in this cell type.
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  • 44
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    Cell & tissue research 279 (1994), S. 221-231 
    ISSN: 1432-0878
    Keywords: Key words: Cytokeratins ; Thymus ; Immunohistochemistry ; Hassal's corpuscles ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Cytokeratin expression in normal postnatal human thymus was studied immunohistochemically by using monoclonal antibodies against various cytokeratin polypeptides. An attempt was made to characterize cell populations giving rise to the cornified structures of Hassal's corpuscles. Monoclonal antibody KB-37, a marker of squamous epithelium basal cells, was applied to distinguish the earliest cells capable of undergoing squamous differentiation. Parts of the subcapsular epithelium were extensively stained with this reagent. This epithelium, like the basal layer of certain squamous epithelia, exibited a high incidence of cytokeratins 13 and 14, and pronounced expression of cytokeratin 19. Simple epithelium cytokeratins 8, 18, and 19 were present in the cortex. Scattered cells reacted with KB-37 antibody. All stellate epithelial cells in the medulla were positive for cytokeratin 19. Most of the medullar epithelial cells were positive for cytokeratins 13, 14 and 17 of complex epithelium, in contrast to the cortex, where only a few cells were positive for these cytokeratins. A significant proportion of the medullar cells was positive for KB-37 antigen. Cytokeratins 8 and 18 were expressed in single cells and in groups of cells surrounding Hassal's corpuscles. The outermost cells of these corpuscles were positive for cytokeratin 19 and KB-37. In the peripheral parts of Hassal's corpuscles, simple epithelium cytokeratins 7, 8, 18, and cytokeratins 4, 13, 14, and 17, characteristic of stratified nonkeratinizing epithelia, were coexpressed with keratinization-specific cytokeratins 10/11. The inner parts of the swirls were uniformly positive for cytokeratins 10/11. However, the expression of other cytokeratins was reduced.
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  • 45
    ISSN: 1432-0878
    Keywords: Enteric nervous system ; Submucosal plexuses ; Myenteric plexus ; Neuropeptides ; Immunohistochemistry ; Intestine, small ; Horse
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The architecture and neurochemistry of the enteric nervous system was studied by use of whole-mount preparations obtained by microdissection of the horse jejunum. A myenteric plexus and two plexuses within the submucosa were identified. The external submucosal plexus lying in the outermost region of the submucosa had both neural and vascular connections with the inner submucosal plexus situated closer to the mucosa. Counts of neurones stained for NADH-diaphorase demonstrated the wide variation in size, shape and neurone content of individual ganglia in both the external and internal submucosal plexuses. The average number of cells/ganglion was similar in each plexus (about 25 cells). Immunoreactivities for galanin, vasoactive intestinal peptide and neuropeptide Y were observed in nerve cell bodies and fibres of each of the plexuses. Immunoreactivity for substance P was extensive and strong in nerve fibres of all plexuses but was weaker in cell bodies of the submucosal neurones and absent in the cell bodies of the myenteric plexus. Comparative quantitative analysis of immunoreactive cell populations with total cell numbers (enzyme staining) was indicative of neuropeptide colocalization in the external submucosal plexus.
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  • 46
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    Cell & tissue research 278 (1994), S. 57-64 
    ISSN: 1432-0878
    Keywords: Calbindin ; Sensory nerve endings ; Esophagus ; Immunohistochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Immunoreactivity for calbindin was found in nerve endings with irregular laminar shapes in the rat esophagus. In the myenteric ganglia, laminar endings of a range of sizes formed a complex network and appeared to lie at the surface of the ganglion. The myenteric ganglia that contained nerve endings were most abundant in the upper portion of the eosphagus, their number decreasing orally to anally. Calbindin-immunoreactive nerve cell bodies were scattered throughout the esophagus. Laminar terminals were found in the connective tissue of the lamina propria immediately beneath the epithelium and in the muscularis mucosae. Occasional nerve branches formed a network of aborizing endings that surrounded part of the submucosal arterioles. Immunoreactive nerve endings in the mucosa and submucosa were present only in the upper part of the cervical esophagus. Unilateral vagotomy caused a remarkable decrease in the number of the myenteric ganglia containing the calbindin-immunoreactive laminar endings after 15 days or survival; in some of ganglia, the laminar structures disappeared and nerve endings showing weak immunoreactivity had an indistinct appearance, so that the outline of the ganglia became obscure. In operated rats at 24 days, the number of innervated ganglia was about half that in normal rats. However, there was no change in the morphology and the occurrence of the immunoreactive laminar structures in the mucosa and submucosa after denervation. The results show that many of the laminar endings that are immunoreactive for calbindin in the myenteric ganglia are derived from the vagus nerve. Thus, the calbindin-immunoreactive nerve endings with laminar expansions that are found in the rat eosphageal wall could be sensory receptors.
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  • 47
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    Cell & tissue research 278 (1994), S. 171-175 
    ISSN: 1432-0878
    Keywords: Tachykinin ; Substance P ; Sinus venosus ; Heart ; Immunohistochemistry ; Dogfish, Scyliorhinus canicula (Elasmobranchii)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The sinus venosus of the elasmobranch heart is characterized by the presence of large bundles of unmyelinated nerve fibres that bulge into the cardiac lumen, below the endocardium. In the dogfish (Scyliorhinus canicula), these fibres contain numerous dense-core membrane-bounded granules of about 200 nm in diameter. Most intramural ganglion cells of the sinus venosus also show densely packed granules similar to those found in the subendocardial fibres. We have observed strong substance-P-like immunoreactivity in the large fibre bundles and in the perikarya of the ganglion cells. Preabsorption of the antisera with fragment 7–11 of substance P has shown that the antisera recognize the tachykinin canonic sequence. Our findings suggest that an undetermined tachykinin is secreted in the elasmobranch heart, and that it is probably released into the blood stream in the context of a little-known neuroendocrine system.
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  • 48
    ISSN: 1432-0878
    Keywords: Carotid body ; Chief cells ; Catecholamine ; Serotonin ; γ-Aminobutyric acid ; Immunohistochemistry ; Mouse (BALB/c)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The immunohistochemical study revealed tyrosine hydroxylase (TH), dopamine β-hydroxylase (DBH), phenylethanolamine N-methyltransferase (PNMT), serotonin, glutamate decarboxylase (GAD) and γ-aminobutyric acid (GABA) immunoreactivities in the mouse carotid body. TH and DBH immunoreactivities were found in almost all chief cells and a few ganglion cells, and in relatively numerous varicose nerve fibers of the carotid body. The histofluorescence microscopy showed catecholamine fluorescence in almost all chief cells. However, no PNMT immunoreactivity was observed in the carotid body. Serotonin, GAD and GABA immunoreactivities were also seen in almost all chief cells of the carotid body. From combined immunohistochemistry and fluorescence histochemistry, catecholamine and serotonin or catecholamine and GABA were colocalized in almost all chief cells. Thus, these findings suggest that noradrenaline, serotonin and GABA may be synthesized and co-exist in almost all chief cells of the mouse carotid body and may play roles in chemoreceptive functions.
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  • 49
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    Cell & tissue research 278 (1994), S. 379-387 
    ISSN: 1432-0878
    Keywords: Key words: Enteric nervous system ; Immunohistochemistry ; 5-Hydroxytryptamine ; Colon ; Guinea-pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The presence of 5-hydroxytryptamine in enteric neurons of the guinea-pig distal colon was demonstrated by immunohistochemistry and the projections of the neurons were determined. 5-Hydroxytryptamine-containing nerve cells were observed in the myenteric plexus but no reactive nerve cells were found in submucous ganglia. Varicose reactive nerve fibres were numerous in the ganglia of both the myenteric and submucous plexuses, but were infrequent in the longitudinal muscle, circular muscle, muscularis mucosae and mucosa. Reactivity also occurred in enterochromaffin cells. Lesion studies showed that the axons of myenteric neurons projected anally to provide innervation to the circular muscle and submucosa and to other more anally located myenteric ganglia. The results suggest that a major population of 5- hydroxytryptamine neurons in the colon is descending interneurons, most of which extend for 10 to 15 mm in the myenteric plexus and innervate both 5-hydroxytryptamine and non-5-hydroxytryptamine neurons.
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  • 50
    ISSN: 1432-0878
    Keywords: Catecholamine synthesizing enzymes ; Neuropeptide Y ; Vasoactive intestinal polypeptide ; Ganglion cells ; Adrenal gland ; Immunohistochemistry ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Immunohistochemistry has been used to demonstrate tyrosine hydroxylase (TH), dopamine-β-hydroxylase (DBH), phenylethanolamine N-methyltransferase (PNMT), neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP) immunoreactivities, and acetylcholinesterase (AChE) activity was demonstrated in rat adrenal glands. The TH, DBH, NPY and VIP immunoreactivities and AChE activity were observed in both the large ganglion cells and the small chromaffin cells whereas PNMT immunoreactivity was found only in chromaffin cells, and not in ganglion cells. Most intraadrenal ganglion cells showed NPY immunoreactivity and a few were VIP immunoreactive. Numerous NPY-immunoreactive ganglion cells were also immunoreactive for TH and DBH; these cells were localized as single cells or groups of several cells in the adrenal cortex and medulla. Use of serial sections, or double and triple staining techniques, showed that all TH- and DBH-immunoreactive ganglion cells also showed NPY immunoreactivity, whereas some NPY-immunoreactive ganglion cells were TH and DBH immunonegative. NPY-immunoreactive ganglion cells showed no VIP immunoreactivity. AChE activity was seen in VIP-immunopositive and VIP-immunonegative ganglion cells. These results suggest that ganglion cells containing noradrenaline and NPY, or NPY only, or VIP and acetylcholine occur in the rat adrenal gland; they may project within the adrenal gland or to other target organs. TH, DBH, NPY, and VIP were colocalized in numerous immunoreactive nerve fibres, which were distributed in the superficial adrenal cortex, while TH-, DBH- and NPY-immunoreactive ganglion cells and nerve fibres were different from VIP-immunoreactive ganglion cells and nerve fibres in the medulla. This suggests that the immunoreactive nerve fibres in the superficial cortex may be mainly extrinsic in origin and may be different from those in the medulla.
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  • 51
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    Cell & tissue research 275 (1994), S. 143-156 
    ISSN: 1432-0878
    Keywords: Larynx ; Trachea ; Endocrine cells ; Neuroepithelial bodies ; Immunohistochemistry ; Regulatory peptides ; Serotonin (5-HT) ; Golden hamster
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The ontogeny of protein gene product 9.5 (PGP 9.5), serotonin (5-HT), calcitonin gene-related peptide (CGRP), and calcitonin (CT) immunoreactivity was evaluated in small-granule endocrine cells of hamster laryngotracheal epithelium from fetal day 11 to adulthood. Two centrifugal (proximal-to-distal) patterns of differentiation occur. The first pattern begins during fetal life. Endocrine cells, single and clustered in groups (presumptive-or protoneuroepithelial bodies, pNEBs), initially colocalize immunostaining for PGP 9.5, 5-HT, and CGRP in the larynx and proximal 2/3 of the trachea on day 12 and spread to the caudal trachea on day 13.5-HT disappears fleetingly during the 24 h preceding birth; other-wise immunoreactivity for all three substances persists into adulthood. The clusters of endocrine cells survive beyond birth but are so diluted by expansion of the nonendocrine epithelium as to become inconspicuous. Since innervation was not actually observed, these clusters may persist as pNEBs, without developing connections to afferent or efferent nerve fibers. The second pattern concerns single small-granule cells stainable for CGRP but not for 5-HT. These cells first appear in the larynx and cartilaginous part of the cranial trachea on postnatal day 3, and in the middle and caudal trachea, on day 5. The cells increase in number on day 7. In adults, they predominate among endocrine cells of the cartilaginous region. A subset of these cells begins to co-express CT proximally on postnatal day 10, reaching the caudal end of the trachea by 3 weeks. A few elements of the older 5-HT-positive population may also become immunoreactive for CT in juvenile hamsters.
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  • 52
    ISSN: 1432-0878
    Keywords: FSH ; Immunohistochemistry ; Receptor mRNA ; In situ hybridization ; Sertoli cell ; Testis ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Testicular biopsies from 82 oligo-or azoospermic male patients were subjected to immunostaining using anti-human FSH antibodies. Histological evaluation showed normal spermatogenesis (nspg) in 7 (FSH: 2.7±0.7), mixed atrophy (ma) in 63 (FSH:5.3±0.5), and bilateral or unilateral Sertoli Cell Only syndrome (SCO) in 12 (FSH:21.7±3.5) patients. For the relationship between FSH values and testicular histology, see Bergmann et al. (1994). FSH immunoreactivity was found exclusively in Sertoli cells and in some interstitial cells. Seminiferous epithelium showing normal or impaired spermatogenesis displayed only weak immunoreactivity compared to intense immunoreaction, i.e. large and numerous vesicles in Sertoli cells of SCO tubules in biopsies showing mixed atrophy or SCO. In addition, h-FSH receptor mRNA was demonstrated by in situ hydridization using biotinylated cDNA antisense oligonucleotides. Hybridization signals were found within the seminiferous epithelium exclusively in Sertoli cell cytoplasm associated with normal spermatogenesis and in epithelia showing different signs of impairment, including SCO. It is concluded that: (1) Sertoli cells are the only cells within the seminiferous epithelium expressing FSH receptors; (2) the accumulation of FSH immunoreactivity in Sertoli cells of SCO tubules appears to be a sign of impaired Sertoli cell function.
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  • 53
    ISSN: 1432-0878
    Keywords: Adrenal ; Autonomic nervous system ; Schwann cells ; Tyrosine hydroxylase ; GAP-43 ; Electron microscopy ; Immunohistochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We have localized at light and electron-microscopic level the growth-associated protein GAP-43 in adrenal gland using single and double labelling immunocytochemistry. Clusters of GAP-43-immunofluorescent chromaffin cells and many immunofluorescent fibres were observed in the medulla. GAP-43-immunoreactive fibres also formed a plexus under the capsule, crossed the cortex and ramified in the zona reticulata. Double labelled sections showed the coexpression of GAP-43 with a subpopulation of tyrosine hydroxylase-and of dopamine-β-hydroxylase-immunoreactive chromaffin cells. Dual colour immunofluorescence for GAP-43 and calcitonin gene-related peptide (CGRP) revealed that some of the GAP-43-immunoreactive fibres also express CGRP. Pre-embedding electron microscopy showed GAP-43 immunoreactivity associated with the plasma membranes and cytoplasm of noradrenaline-producing chromaffin cells, and with processes of nonmyelin-forming Schwann cells. Immunoreactive unmyelinated axons and terminals were also observed. The immunostained terminals made symmetrical synaptic contacts with chromaffin cells. Immunoreactive unmyelinated fibres and small terminals were present in the cortex. Our results show that GAP-43 is expressed in noradrenergic chromaffin cells and in various types of nerve fibres that innervate the adrenal. Likely origins for these fibres include preganglionic sympathetic fibres which innervate chromaffin cells, postganglionic sympathetic fibres in the cortex, and CGRP containing sensory fibres.
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  • 54
    ISSN: 1432-0878
    Keywords: Nervous system, insect ; Octopamine ; DUM neurons ; Immunohistochemistry ; Accessory glands ; Periplaneta americana (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The musculature of the mushroom-shaped accessory gland receives innervation from trunks 5C1 of the phallic nerves, which arise from the posterior part of the terminal abdominal ganglion of the male cockroach Periplaneta americana. Anterograde cobalt filling through trunks 5C1 with the subsequent precipitating procedure has shown the fine innervation of the accessory gland. By retrograde cobalt filling through the same trunks, different types of cells have been mapped in the terminal abdominal ganglion. About 25 dorsal unpaired median (DUM) neurons have been identified among them. About 36 octopamine-like immunoreactive DUM neurons with large somata have been characterized in whole-mount preparations of the terminal abdominal ganglion. The combination of the cobalt-filling technique with immunohistochemical mapping of cells suggests an octopaminergic innervation of the musculature of the accessory gland by DUM neurons.
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  • 55
    ISSN: 1432-0878
    Keywords: Key words: β-Galactoside-binding lectin ; Dermis ; Skin ; Chick embryo ; Immunohistochemistry ; Keratinization ; Mucous metaplasia ; Domestic fowl
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. In order to elucidate the roles of metal-independent animal lectins, we systematically investigated changes in expression of 2 kinds of β-galactoside-binding isolectins (MW 14 and 16 kDa) in the dermis of chick embryonic tarsometatarsal skin during the course of development. These lectins were immunohistochemically located at different stages of development both in ovo and in vitro by light and electron microscopy. Light- microscopic observation showed that while positive staining for the 14-kDa lectin was weak at days 8 and 10 it became intense after day 13. In contrast, staining for the 16-kDa lectin was intense at days 8, 10, and 13, but it became weak after day 17 when keratinization of the epidermis was completed. Immuno-electron-microscopic observation revealed that both the 14 and 16-kDa lectins were located on the basement membrane, in the extracellular matrix, and in both the cytoplasm and the nucleus of dermal fibroblasts. Distribution of the 2 isolectins was also examined in cultured skin explants in vitro. The results were almost the same as those obtained in ovo when the skin explant was keratinized in the presence of hydrocortisone. However, in the skin explant where keratinization was prevented and mucous metaplasia was induced by the addition of vitamin A, the distribution of the 14-kDa lectin in the epidermis was significantly affected. These results indicate that (1) the expression of the 2 isolectins is differently regulated in both the dermis and epidermis, (2) the 16-kDa lectin is involved in the early stage of the formation of the dermis and the basement membrane and is replaced by the 14-kDa lectin as keratinization of the epidermis occurs, and (3) the expression of the 2 isolectins in the dermis is not significantly affected by the induction of mucous metaplasia, in contrast to their drastic changes in the epidermis.
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  • 56
    ISSN: 1432-0878
    Keywords: Brain mapping ; GABA ; Immunohistochemistry ; Visual reflexes ; Salamanders, Pleurodeles waltli, Triturus alpestris (Urodela)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The distribution of GABAergic neurons in brains of the family Salamandridae (Pleurodeles waltli, Triturus alpestris) has been investigated immunohistochemically with an antibody against gamma-aminobutyric acid (GABA). In adult animals, immunoreactive neurons, fibers, and terminals are abundantly labeled. In the telencephalon, pallial areas contain fewer GABAergic neurons and fibers than basal forebrain areas. The amygdalar complex and the habenulae have a complex pattern of GABA-immunoreactivity that is especially pronounced within the neuropil. The pretectal and basal optic systems are provided with GABAergic neurons, corroborating electrophysiological results. The dorsal thalamus and parts of the torus semicircularis are almost completely devoid of GABA-immunoreactive neurons. In the torus, magnocellular neurons known to project to the contralateral counterpart are distinctly GABA-immunoreactive. During ontogeny, GABAergic neurons arise early when the first reflexive movements occur after mechanical stimulation. At stage 28, cells are labeled initially near the nucleus of the medial longitudinal fasciculus, which is the first supraspinal tract to appear in ontogeny. At stage 30 (still before hatching), GABAergic neurons are found in the pretectum, immunoreactive neurons arising in the dorsal tegmentum slightly later. Both systems are known to mediate basic reflexes in gaze stabilization. The commissura posterior is GABAergic at early stages suggesting an important functional role in homonymous inhibition between both sides. Thus in salamanders, the neurotransmitter GABA displays a complex distribution, similar to that in other vertrebrates. This pattern emerges early in ontogeny.
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  • 57
    ISSN: 1432-0878
    Keywords: EGF ; Cartilage ; Growth plate ; Hypophysectomy ; Growth hormone ; Immunohistochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Epiphyseal growth plate cartilages from the proximal tibia of normal, hypophysectomized, and growth hormone (GH)-treated hypophysectomized rats were subjected to immunohistochemistry for detection of epidermal growth factor (EGF). In the normal growth plate, EGF was distributed mainly in the proliferative zone. Hypophysectomy resulted in considerable atrophy of the chondrocytes and the cartilage matrix (a decreased number of mature-type chondrocytes and a decreased ratio of proliferating to hypertrophic chondrocytes) and a significant diminution of EGF immunoreactivity. Treatment with GH reversed these effects of hypophysectomy, causing an increased thickness of the growth plate and EGF-reactive sites in all chondrocyte layers. The most intense immunostaining for EGF, however, was frequently seen in the nuclei of chondrocytes with flattened appearance. It appears that EGF could be incorporated or synthesized in chondrocytes having marked mitogenic activity. The present results, taken with previous data on EGF involvement in growth of cartilaginous tissue in vivo and in vitro, strongly suggest that EGF-immunoreactive chondrocytes are involved in cartilage proliferation and growth under the specific influence of GH.
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  • 58
    ISSN: 1432-0878
    Keywords: Placenta ; Protease inhibitors ; Immunohistochemistry ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Proteases and their inhibitors play a pivotal role in developmental and differentiative processes. In the present report we investigated the immunohistochemical localization of α1-antitrypsin, α1-antichymotrypsin and inter-α-trypsin inhibitor in first trimester as well as in term human placentas. For this purpose polyclonal antibodies against these serine-protease inhibitors were used. All inhibitors were expressed in the villous syncytiotrophoblast of first and last trimester placentas. Placental fibrinoid was positively stained for α1-antitrypsin and inter-α-trypsin inhibitor throughout gestation. α1-Antitrypsin and α1-antichymotrypsin showed a strong immunostaining in the Hofbauer cells (first trimester and full term placentas). Extravillous cytotrophoblast was negative for the three protease inhibitors throughout gestation. The presence of the three inhibitors in the syncytiotrophoblast suggests a role in coagulative, invasive and immunomodulatory processes. Fibrinoid, staining for α1-antitrypsin and inter-α-trypsin inhibitor, could also have an important immunoprotective function. The presence of protease inhibitors in the Hofbauer cells suggests an involvement of these cells in villous remodelling and differentiative processes.
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  • 59
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    Cell & tissue research 278 (1994), S. 379-387 
    ISSN: 1432-0878
    Keywords: Enteric nervous system ; Immunohistochemistry ; 5-Hydroxytryptamine ; Colon ; Guinea-pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The presence of 5-hydroxytryptamine in enteric neurons of the guinea-pig distal colon was demonstrated by immunohistochemistry and the projections of the neurons were determined. 5-Hydroxytryptamine-containing nerve cells were observed in the myenteric plexus but no reactive nerve cells were found in submucous ganglia. Varicose reactive nerve fibres were numerous in the ganglia of both the myenteric and submucous plexuses, but were infrequent in the longitudinal muscle, circular muscle, muscularis mucosae and mucosa. Reactivity also occurred in enterochromaffin cells. Lesion studies showed that the axons of myenteric neurons projected anally to provide innervation to the circular muscle and submucosa and to other more anally located myenteric ganglia. The results suggest that a major population of 5-hydroxytryptamine neurons in the colon is descending interneurons, most of which extend for 10 to 15 mm in the myenteric plexus and innervate both 5-hydroxytryptamine and non-5-hydroxytryptamine neurons.
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  • 60
    ISSN: 1432-0878
    Keywords: Key words: Macrophages ; Adrenal cortex ; Chromaffin cells ; Immunohistochemistry ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. There is increasing evidence for an immune-adrenal interaction in which macrophages may play an important role. However, few data are available with respect to a human intra-adrenal macrophage system. In this study, we have investigated the density, distribution and phenotype of human adrenal macrophages using monoclonal antibodies. Macrophages are localized in all zones of the adrenal gland. These cells exhibit the phenotype of the phagocytotic macrophage compartment (CD11c+, KiM8+). At the ultrastructural level, macrophages are frequently attached to the endothelial wall, but also lie in direct contact with cortical and chromaffin cells. This investigation reveals the cellular basis for the possible role of macrophages in the local immune-neuroendocrine axis.
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  • 61
    ISSN: 1432-0878
    Keywords: Macrophages ; Adrenal cortex ; Chromaffin cells ; Immunohistochemistry ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract There is increasing evidence for an immune-adrenal interaction in which macrophages may play an important role. However, few data are available with respect to a human intra-adrenal macrophage system. In this study, we have investigated the density, distribution and phenotype of human adrenal macrophages using monoclonal antibodies. Macrophages are localized in all zones of the adrenal gland. These cells exhibit the phenotype of the phagocytotic macrophage compartment (CD11c+, KiM8+). At the ultrastructural level, macrophages are frequently attached to the endothelial wall, but also lie in direct contact with cortical and chromaffin cells. This investigation reveals the cellular basis for the possible role of macrophages in the local immune-neuroendocrine axis.
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  • 62
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    Journal of applied phycology 6 (1994), S. 199-210 
    ISSN: 1573-5176
    Keywords: cyanobacteria ; iron deficiency ; photosynthesis ; siderophores ; flavodoxin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Iron is an essential component of electron transport in almost all living organisms. It is particularly important to phototrophs like cyanobacteria because 22–23 irons are required for a complete functional photosynthetic apparatus. Since the low solubility of Fe+++ above neutral pH in oxic ecosystems severely limits the biological availability of iron to aquatic microorganisms, cyanobacteria and other microbes have developed a number of responses to cope with iron deficiency. Cyanobacterial responses to iron stress include the synthesis of an efficient, siderophore-based system to scavenge iron and the substitution of ferredoxin with flavodoxin. An additional response in cyanobacteria involves the alteration of the light-harvesting apparatus that includes the appearance of a new, iron-stress-induced, photosystem II, chlorophyll-binding protein. Although cytochromec-553 has a potential non-iron-containing replacement in plastocyanin, a copper-containing protein, iron stress appears to favor the utilization of cytochromec-553 because siderophores also bind copper and form a complex that is excluded from the cell. This paper is intended primarily as a review of molecular and physiological responses of actively growing cyanobacterial cultures to conditions of iron stress, where iron is present but essentially insoluble, and to differentiate these responses from iron starvation, where the amount of iron in the system is not sufficient for cell growth.
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  • 63
    ISSN: 1573-5176
    Keywords: fluorescence ; photoinhibition ; photosynthesis ; Spirulina ; temperature
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Chlorophyll fluorescence measurements were used to evaluate the effect of temperature on photoinhibition inSpirulina platensis cultures grown in tubular reactors outdoors. Cultures grown at 35 °C during the day time showed a lower reduction in the Fv/Fm ratio as compared to cultures grown at 25 °C. It is demonstrated that the lower temperature photoinhibited cells can undergo a complete recovery once transferred to low light and higher temperature. This recovery does not take place when 100 µg ml-1 chloramphenicol is added to cells. The recovery is light dependent and cells incubated in the dark at low temperature do not show a recovery in the Fv/Fm ratio. The data presented strongly support the hypothesis that photoinhibition takes place in outdoorSpirulina cultures. At the same time it is demonstrated that fluorescence measurements can be used as a fast reliable indication for photoinhibition in outdoor algal cultures.
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  • 64
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    Journal of applied phycology 6 (1994), S. 45-60 
    ISSN: 1573-5176
    Keywords: biofilter ; biofuels ; CO2 ; macroalgae ; marine biomass ; photosynthesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Biomass production from macroalgae has been viewed as important mainly because of the need for pollution abatement. Environmental considerations will increasingly determine product and process acceptability and drive the next generation of economic opportunity. Some countries, including Japan, are actively promoting "green" technologies that will be in demand worldwide in the coming decades. Should an international agreement on CO2-reduction be ratified, its effective use for energy production would be of high priority. This report shows that macroalgae have great potential for biomass production and CO2 bioremediation. Macroalgae have high productivity, as great or greater than the most productive land plants, and do not compete with terrestrial crops for farm land. The review focuses on recent data on productivity, photosynthesis, nutrient dynamics, optimization and economics. Biomass from macroalgae promises to provide environmentally and economically feasible alternatives to fossil fuels. Nevertheless, the techniques and technologies for growing macroalgae on a large-scale and for converting feedstocks to energy carriers must be more fully developed.
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  • 65
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    Journal of applied phycology 6 (1994), S. 309-313 
    ISSN: 1573-5176
    Keywords: light source ; emission spectrum ; photosynthesis ; algae ; absorption ; efficiency
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A method for quantitative evaluation of light sources from the point of their suitability for algal cultivation is described. Two parameters are used for the evaluation: (1) amount of photons emitted in the spectral region absorbed by algae per consumed unit of electrical energy; (2) fraction of photons intercept by the algae that got effectively transformed into chemical energy. The value of parameter (2) depends on the emission spectrum of the light source, the absorption spectrum of the algae, quantum capacity of the algae and on conditions of cultivation. The method permits the assessment of the differences in algal productivity under light of various light sources with respect to other recent methods for comparison.
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  • 66
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    Journal of applied phycology 6 (1994), S. 331-335 
    ISSN: 1573-5176
    Keywords: turbulence ; photosynthesis ; Chlorella ; light/dark cycles ; mass transfer rates
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In mass algal cultures, some form of agitation is usually provided; among other effects, this moves the organisms though an optically dense profile and provides mixing. During this transport, medium frequency fluctuations in the light energy supply are perceived by the algae, which are of the order of 1 Hz and less. It has been suggested that turbulence with the resultant light/dark cycles of medium frequency enhances productivity. However, turbulence has two major influences in a well mixed system: it facilitates fluctuating light regimes and increases the transfer rates between the growth medium and the cultured organism. An estimation of productivity as oxygen liberation was measured under laminar and turbulent flow rates, and varying light/dark ratios. Increased turbulence, which increased exchange rates of nutrients and metabolites between the cells and their growth medium, together with increased light/dark frequencies, increased productivity and photosynthetic efficiency.
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  • 67
    ISSN: 1573-5036
    Keywords: photosynthesis ; osmotic adjustment ; stomatal conductance ; relative water content ; water potential ; water relations
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Four pigeon pea [Cajanus cajan (L.) Mill sp.] cultivars were studied during two cycles of development of water stress and recovery. During these two cycles the genetic variation in vegetative development, leaf water potential, relative water content, photosynthesis, and stomatal conductance was followed. Plants were grown in a greenhouse and irrigated every seventh day. On two occasions (30 days and 80 days from sowing) water was withheld for 13 days in one group of plants and 16 days in another. Control plants were irrigated as usual. The four cultivars differed in their response to drought, with ICPL 215 being the most tolerant cultivar. In the plants exposed to the 13-day water stress, the first stress cycle resulted in preconditioning of plants such that higher values for carbon dioxide exchange rate and relative water content were observed in the second stress cycle. The longer stress period (16 days) resulted in some damage to the photosynthetic capacity and in a slow recovery rate in both cycles. During the second drought cycle the plants retained a water content above the critical value, possibly due to osmotic adjustment. It seems that water status parameters, especially relative water content, may serve as indicators of drought tolerance in pigeon pea varieties and may be useful in breeding programs for cultivation of pigeon pea under semiarid conditions.
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  • 68
    ISSN: 1573-5079
    Keywords: light conditions ; theoretical model ; barley leaf ; photosynthesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A theoretical model is presented describing the distortion of chlorophyll fluorescence spectra of a chloroplast or a group of chloroplasts by the effect of fluorescence reabsorption. Model calculations using the experimental data show that the primary reabsorption effect occurs already within one chloroplast and the spectral distortion depends significantly on the excitation regime of the chloroplast. A theoretical dependence of the distortion function, defined as a change in the F(685)/F(735) fluorescence band ratio, on the mean chlorophyll concentration in a chloroplast is predicted for different light excitation regimes. The distortion of measured chlorophyll fluorescence spectra at 77 K of chloroplast suspension adsorbed on filter papers of two strongly different diffusive reflectivities and at different mean chlorophyll concentrations are discussed with the help of the presented theory.
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  • 69
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    Photosynthesis research 39 (1994), S. 475-489 
    ISSN: 1573-5079
    Keywords: DNA repair ; flavonoids ; gene expression ; oxidative stress ; photosynthesis ; promoter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Influx of solar UV-B radiation (280–320 nm) will probably increase in the future due to depletion of stratospheric ozone. In plants, there are several targets for the deleterious UV-B radiation, especially the chloroplast. This review summarizes the early effects and responses of low doses of UV-B at the molecular level. The DNA molecules of the plant cells are damaged by UV due to the formation of different photoproducts, such as pyrimidine dimers, which in turn can be combatted by specialized photoreactivating enzyme systems. In the chloroplast, the integrity of the thylakoid membrane seems to be much more sensitive than the activities of the photosynthetic components bound within. However, the decrease of mRNA transcripts for the photosynthetic complexes and other chloroplast proteins are among very early events of UV-B damage, as well as protein synthesis. Other genes, encoding defence-related enzymes, e.g., of the flavonoid biosynthetic pathway, are rapidly up-regulated after commencement of UV-B exposure. Some of the cis-acting nucleotide elements and trans-acting protein factors needed to regulate the UV-induced expression of the parsley chalcone synthase gene are known.
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  • 70
    ISSN: 1573-5079
    Keywords: photosynthesis ; electrometry ; membrane potential ; electron transfer ; charge displacement ; electrostatic ; Chloroflexus aurantiacus ; Rhodopseudomonas viridis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The thermophilic phototrophChloroflexus aurantiacus possesses a photosynthetic reaction center (RC) containing a pair of menaquinones as primary (QA) and secondary (QB) electron acceptors and a bacteriochlorophyll dimer (P) as a primary donor. A tetraheme cytochromec 554 with two high(H)- and two low(L)-potential hemes operates as an immediate electron donor for P. The following equilibrium Em,7 values were determined by ESR for the hemes in whole membrane preparations: 280 mV (H1), 150 mV (H2), 95 mV (L1) and 0 mV (L2) (Van Vliet et al. (1991) Eur. J. Biochem. 199: 317–323). Partial electrogenic reactions induced by a laser flash inChl. aurantiacus chromatophores adsorbed to a phospholipid-impregnated collodion film were studied electrometrically at pH 8.3. The photoelectric response included a fast phase of ΔΨ generation (τ 〈 10 ns, phase A). It was ascribed to the charge separation between P+ and QA − as its amplitude decreased both at high and low Eh values (Em,high=360±10 mV, estimated Em,low∼\s-160 mV) in good agreement with Em values for P/P+ and QA/QA − redox couples. A slower kinetic component appeared upon reduction of the cytochromec 554 hemes (phase C). With H1 reduced before the flash the amplitude of phase C was equal to 15–20% of that of phase A and its rise time was 1.2–1.3 μs: we attribute this phase to the electrogenic electron transfer from H1 to P+. Pre-reduction of H2 decreased the τ value to about 700–800 ns and increased the amplitude of phase C to 30–35% of that of phase A. Pre-reduction of L1 further accelerated phase C (up to τ of 500 ns) and induced a reverse electrogenic phase with τ of 12 μs and amplitude equal to 10% of phase A. Upon pre-reduction of L2 the rise time of phase C was decreased to about 300 ns and its amplitude decreased by 30%. The acceleration in the onset of phase C is explained by the acceleration of the rate-limiting H1 ⇒ P electrogenic reaction after reduction of the other hemes due to their electrostatic influence; a P-H1-(L1-L2)-H2 alignment of redox centers with an approximately rhombic arrangement of the cytochromec 554 hemes is proposed. The observed reverse phase is ascribed to the post-flash charge redistribution between the hemes. Redox titration of the amplitude of phase C yielded the Em,8.3 values of H1, H2 and L2 hemes: 340±10 mV for H1, 160±20 mV for H2 and −40±40 mV for L2.
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  • 71
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    Photosynthesis research 41 (1994), S. 175-180 
    ISSN: 1573-5079
    Keywords: Chlorobium ; Chloroflexus ; chlorophyll ; light-harvesting ; photosynthesis ; resonance Raman
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Resonance Raman experiments were performed on different green bacteria. With blue excitation, i.e. under Soret resonance or preresonance conditions, resonance Raman contributions were essentially arising from the chlorosome pigments. By comparing these spectra and those of isolated chlorosomes, it is possible to evaluate how the latter retain their native structure during the isolation procedures. The structure of bacteriochlorophyll oligomers in chlorosomes was interspecifically compared, in bacteriochlorophyllc- and bacteriochlorophylle- synthesising bacteria. It appears that interactions assumed by the 9-keto carbonyl group are identical inChlorobium limicola, Chlorobium tepidum, andChlorobium phaeobacteroides. In the latter strain, the 3-formyl carbonyl group of bacteriochlorophylle is kept free from intermolecular interactions. By contrast, resonance Raman spectra unambiguously indicate that the structure of bacteriochlorophyll oligomers is slightly different in chlorosomes fromChloroflexus auranticus, either isolated or in the whole bacteria.
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  • 72
    ISSN: 1573-5079
    Keywords: bacteriochlorophyll ; chlorosomeChlorobium limicola ; green bacteria ; photosynthesis ; monogalactosyl diglyceride
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Artificial aggregates of bacteriochlorophyllc (BChlc) were formed in an aqueous medium in the presence of a lipid, monogalactosyl diglyceride (MGDG), and the optical properties of those aggregates were studied by absorption and circular dichroism (CD) mainly. Four BChlc homologs, ([E,E]BChlc F, [P,E]BChlc F, [E,M]BChlc F and [I,E]BChlc F), were isolated from the green photosynthetic bacteriumChlorobium limicola strain 6230. Above 0.0004%, MGDG induced a red-shift of the absorption maxima of BChlc aggregates. At 0.003% MGDG BChlc aggregates showed absorption maxima in the range of 724 to 745 (±3) nm with a shift of 12 to 24 (±3) nm depending on the homolog species. Four kinds of BChlc-MGDG aggregates showed characteristic CD spectra. [E,M]BChlc F gave rise to a CD spectrum similar to that of chlorosomes, while the other three gave spectra of opposite sign. These aggregates are sensitive to 1-hexanol treatment; in a saturating amount (0.85%) of 1-hexanol, all the homologs gave a monomer-like absorption spectrum peaking at 670nm. At an intermediate concentration (0.5%), [E,M]BChlc F showed an enhanced CD intensity, as observed in native chlorosomes. Resonance Raman spectra of the monomer-like BChlc samples indicated that the keto vibrational band at ca. 1640 cm−1 was considerably weakened by the 0.85% 1-hexanol treatment, however the 1680 cm−1 band characteristic of a free keto group did not appear. These results indicate that the artificial aggregates formed by purified BChlc homologs and MGDG are good models for studying chlorosomes structure.
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  • 73
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    Photosynthesis research 39 (1994), S. 201-204 
    ISSN: 1573-5079
    Keywords: automatic determination ; fluorescence parameters ; fluorometer ; induction curve ; photosynthesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The (Fpl-Fo)/Fv value of the fluorescence induction curve is shown to be a more suitable parameter to detect a wider range of heat stress damage to thylakoid membranes as compared to quantities t 1/2 (time of fluorescence rise from Fo to (Fo+Fm)/2 level) and % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGak0Jf9crFfpeea0xh9v8qiW7rqqrFfpeea0xe9Lq-Jc9% vqaqpepm0xbba9pwe9Q8fs0-yqaqpepae9pg0FirpepeKkFr0xfr-x% fr-xb9adbaqaaeGaciGaaiaabeqaamaabaabaaGcbaWaa0aaaeaacq% aHepaDaaaaaa!39D5!\[\overline \tau \] (the fluorescence induction time defined as the area above the induction curve normalized to Fv=1). A method for exact and automatic Fpl determination is presented. A break point in the quality and behaviour of the fluorescence induction curve of barley leaves incubated at 49°C was reached at the moment (about 240 s) when the transformation of PS II active (QB-reducing) to PS II inactive (QB-non-reducing) centres was completed. The meaning of the standard Fv and Fv/Fm parameter was then changed. The method of Fpl determination described here may help to increase the analytical value of the standard chlorophyll fluorometers.
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  • 74
    ISSN: 1573-5079
    Keywords: chlorosis ; diagnostic ; flavodoxin ; iron-limitation ; photosynthesis ; phytoplankton
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Iron supply has been suggested to influence phytoplankton biomass, growth rate and species composition, as well as primary productivity in both high and low NO3 − surface waters. Recent investigations in the equatorial Pacific suggest that no single factor regulates primary productivity. Rather, an interplay of bottom-up (i.e., ecophysiological) and top-down (i.e., ecological) factors appear to control species composition and growth rates. One goal of biological oceanography is to isolate the effects of single factors from this multiplicity of interactions, and to identify the factors with a disproportionate impact. Unfortunately, our tools, with several notable exceptions, have been largely inadequate to the task. In particular, the standard technique of nutrient addition bioassays cannot be undertaken without introducing artifacts. These so-called ‘bottle effects’ include reducing turbulence, isolating the enclosed sample from nutrient resupply and grazing, trapping the isolated sample at a fixed position within the water column and thus removing it from vertical movement through a light gradient, and exposing the sample to potentially stimulatory or inhibitory substances on the enclosure walls. The problem faced by all users of enrichment experiments is to separate the effects of controlled nutrient additions from uncontrolled changes in other environmental and ecological factors. To overcome these limitations, oceanographers have sought physiological or molecular indices to diagnose nutrient limitation in natural samples. These indices are often based on reductions in the abundance of photosynthetic and other catalysts, or on changes in the efficiency of these catalysts. Reductions in photosynthetic efficiency often accompany nutrient limitation either because of accumulation of damage, or impairment of the ability to synthesize fully functional macromolecular assemblages. Many catalysts involved in electron transfer and reductive biosyntheses contain iron, and the abundances of most of these catalysts decline under iron-limited conditions. Reductions of ferredoxin or cytochrome f content, nitrate assimilation rates, and dinitrogen fixation rates are amongst the diagnostics that have been used to infer iron limitation in some marine systems. An alternative approach to diagnosing iron-limitation uses molecules whose abundance increases in response to iron-limitation. These include cell surface iron-transport proteins, and the electron transfer protein flavodoxin which replaces the Fe-S protein ferredoxin in many Fe-deficient algae and cyanobacteria.
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  • 75
    ISSN: 1573-5079
    Keywords: photosynthesis ; membrane protein ; protein structure ; light harvesting
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have used antibodies generated against synthetic peptides to determine the topology of the 43 kD chlorophyll a binding protein (CP 43) of Photosystem II. Based on the pattern of proteolytic fragments detected (on western blots) by peptide specific antibodies, a six transmembrane span topological model, with the amino and carboxyl termini located on the stromal membrane surface, is predicted. This structure is similar to that predicted for CP 47, a PS II chlorophyll a binding protein (Bricker T (1990) Photosynth Res 24: 1–13). The model is discussed in reference to the possible location of chlorophyll binding sites.
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  • 76
    ISSN: 1573-5036
    Keywords: crop growth ; gas exchange ; minirhizotrons ; nutrients ; photosynthesis ; respiration ; rhizotron ; root observation ; root growth ; water balance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A research facility is described for the integrated study of soil-root-shoot-atmosphere relationships in crops. The Wageningen Rhizolab has been in use since 1990, and consists of two rows, each with eight below-ground compartments aligned along a corridor. A rain shelter automatically covers the experimental area at the start of rainfall. Compartments are 125 cm × 125 cm and 200 cm deep. Each compartment has a separate drip irrigation system. Crop canopy photosynthesis, respiration, and transpiration can be measured simultaneously and continuously on four out of eight compartments at a time. Each compartment can be filled with a selected soil material (repacked soil) and is accessible from the corridor over its full depth. Multiple sensors for measuring soil moisture status, electrical conductivity, temperature, soil respiration, trace gases and oxygen are installed in spatial patterns in accordance with the requirements of the experiments. Sensors are connected to control and data-acquisition devices. Likewise, provisions have been made to sample manually the soil solution and soil atmosphere. Root observation tubes (minirhizotrons) are installed horizontally at depth intervals ranging from 5 cm (upper soil layers) to 25 cm (below 1 m). The facility is at present in use to study growth and development of vegetation (crops) in relation to drought, nutrient status, soil-borne diseases, and underground root competition. One important application is the study of elevated CO2 concentration and climate change and the way they affect crops and their carbon economy. Growth and development of field grown vegetables and winter cover crops are also evaluated. The common aspect of those studies is to gain a better understanding of crop growth under varying environmental conditions, and to collect datasets that may help to improve mechanistic crop growth simulation models that can address suboptimal growth conditions.
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  • 77
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    Photosynthesis research 39 (1994), S. 389-400 
    ISSN: 1573-5079
    Keywords: elevated CO2 ; nitrogen supply ; photosynthesis ; acclimation ; growth
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A common observation in plants grown in elevated CO2 concentration is that the rate of photosynthesis is lower than expected from the dependence of photosynthesis upon CO2 concentration in single leaves of plants grown at present CO2 concentration. Furthermore, it has been suggested that this apparent down regulation of photosynthesis may be larger in leaves of plants at low nitrogen supply than at higher nitrogen supply. However, the available data are rather limited and contradictory. In this paper, particular attention is drawn to the way in which whole plant growth response to N supply constitutes a variable sink strength for carbohydrate usage and how this may affect photosynthesis. The need for further studies of the acclimation of photosynthesis at elevated CO2 in leaves of plants whose N supply has resulted in well-defined growth rate and sink activity is emphasised, and brief consideration is made of how this might be achieved.
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  • 78
    ISSN: 1573-5117
    Keywords: Ruppia drepanensis ; ammonia toxicity ; temperature effects ; photosynthesis ; growth
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In a laboratory experiment, Ruppia drepanensis Tineo seedlings from a brackish marsh in Southern Spain were grown at 20 and 30 °C, at three different nitrogen levels. These levels were obtained by the addition of a slow release fertilizer (23% NH4NO3 by weight) to a sediment mixture of sand and clay (3:1). Several morphometric parameters were recorded during the first five weeks of the experiment, and photosynthesis and respiration were measured after 7 weeks of growth. Results showed a significant reduction of growth and development with increasing nitrogen and temperature levels. Dark respiration increased strongly at high nitrogen levels. At the same time, net photosynthesis at 250 and 500 µE m-2 s-1, Pm, Km and LCP were not affected by either factor. We attribute these phenomena to ammonia toxicity, since relatively high total ammonia (NH3 + NHf4 p+) levels were found in the interstitial water.
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  • 79
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    Hydrobiologia 289 (1994), S. 65-72 
    ISSN: 1573-5117
    Keywords: photosynthesis ; selective environments ; resuspension ; disturbance ; rivers ; shallow lakes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Factors affecting phytoplankton productivity are analysed in turbid systems, such as shallow lakes and rivers. When resuspension from the sediment or loading from the catchment significantly increases inorganic (non-algal) turbidity and hence light attenuation potentials for high production are not realised. Energy available for phytoplankton growth is strongly regulated by underwater light availability which depends on the critical mixing depth, fluctuating light intensities and algal circulation patterns. Higher production rates in shallow waters are often compensated by greater algal respiration due to higher water temperatures when compared to deeper lakes. Total daily integral production of turbulent, turbid environments can be predicted from a combination of easily measured variables such as maximum photosynthetic rates, algal biomass, surface irradiance and some measure of underwater light attenuation.
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  • 80
    ISSN: 1573-5087
    Keywords: homobrassinolide ; irrigated ; membrane stability ; moisture-stress ; nitrate reductase activity ; photosynthesis ; Triticum aestivum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Homobrassinolide (BR) was applied either as a seed treatment or foliar spray to two contrasting wheat varieties, viz. C306 (drought tolerant) and HD2329 (drought susceptible), to examine its effects on plant metabolism and grain yield under irrigated and moisture-stress/rainfed conditions. BR application resulted in increased relative water content, nitrate reductase activity, chlorophyll content and photosynthesis under both conditions. BR application also improved membrane stability (lower injury). These beneficial effects resulted in higher leaf area, biomass production, grain yield and yield related parameters in the treated plants. All the treatments were significantly better than the untreated control. Generally, 0.05 ppm either as a seed treatment or foliar spray was more effective than the 0.01 ppm treatment. The drought-tolerant genotype C306 showed more response to BR application under moisture-stress/rainfed condition than HD 2329. Increased water uptake, membrane stability and higher carbon dioxide and nitrogen assimilation rates under stress seemed to be related to homobrassinolide-induced drought tolerance.
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  • 81
    ISSN: 1573-5117
    Keywords: potamoplankton ; regulated river ; transport of carbon and nutrients ; photosynthesis ; respiration ; dynamics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The ecological importance of the River Meuse phytoplankton with regard to carbon and nutrient transport has been examined in two reaches of the Belgian course of the river. Field measurements of total particulate organic carbon (POC), particulate organic nitrogen (PON) and particulate phosphorus (PP) show that the large autochtonous production of organic matter strongly affects the carbon and nutrient budget of the aquatic system. During the growing season, phytoplankton accounts for nearly 60% of the POC and dominates the PON. Calculations of the carbon and oxygen budget in the upper reach of the Belgian Meuse demonstrates that the ecosystem is autotrophic, i.e. that autochtonous FPOM (fine particulate organic matter) production is the major carbon input. This suggests that in large lowland rivers, primary production (P) may exceed community respiration (R), i.e. P:R〉1, whereas they are assumed to be heterotrophic (P:R〈1) in the River Continuum concept. The question of maintenance of phytoplankton in turbid mixed water columns is also addressed, and the case of the River Meuse is treated on the basis of studies of photosynthesis and respiration (ETS measurements). The results suggest that the potamoplankton may show some low-light acclimation, through an increase of chlorophyll a relative to biomass, when it comes to deep downstream reaches, and that algal respiration rate may be reduced. A simulation of the longitudinal development of the algal biomass shows the different phases of algal growth and decline along the river and brings support to the ‘importation hypothesis’ for explaining maintenance of potamoplankton in the downstream reaches.
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  • 82
    ISSN: 1573-5079
    Keywords: photosynthesis ; ascorbate peroxidase ; Mehler reaction ; cyclic PS I ; chlorophyll fluorescence ; 9-aminoacridine fluorescence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Simultaneous measurements of 9-aminoacridine (9-AA) fluorescence quenching, O2-uptake and chlorophyll fluorescence of intact spinach chloroplasts were carried out to assess the relationship between the transthylakoidal ΔpH and linear electron flux passing through Photosystem II. Three different types of O2-dependent electron flow were investigated: (1) Catalysed by methyl viologen; (2) in the absence of a catalyst and presence of an active ascorbate peroxidase (Mehler-peroxidase reaction); (3) in the absence of a catalyst and with the ascorbate peroxidase being inhibited by KCN (Mehler reaction). The aim of this study was to assess the relative contribution of ΔpH-formation which is not associated with electron flow through Photosystem II and, which should reflect Photosystem I cyclic flow under the different conditions. The relationship between the extent of 9-AA fluorescence quenching and O2-uptake rate was found to be almost linear when methyl viologen was present. In the absence of methyl viologen (Mehler reaction) an increase of 9-AA fluorescence quenching to a value of 20% at low light intensities was associated with considerably less O2-uptake than in the presence of methyl viologen, indicating the involvement of cyclic flow. These findings are in agreement with a preceding study of Kobayashi and Heber (1994). However, when no KCN was added, such that the complete Mehler-peroxidase reaction sequence was operative, the relationship between 9-AA fluorescence quenching and the flux through PS II, as measured via the chlorophyll fluorescence parameter ΔF/Fm′ × PAR, was identical to that observed in the presence of methyl viologen. Under the assumption that methyl viologen prevents cyclic flow, it is concluded that there is no significant contribution of cyclic electron flow to ΔpH-generation in intact spinach chloroplasts.
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  • 83
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    Journal of chemical ecology 20 (1994), S. 969-978 
    ISSN: 1573-1561
    Keywords: Artemisinin ; arteannuic acid ; photosynthesis ; respiration ; chlorophyll ; Lemna minor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The effects of artemisinin and arteannuic acid extracted fromArtemisia annua on the physiology ofLemna minor were evaluated. Changes in frond production, growth, dry weight, and chlorophyll content ofL. minor were determined. Photosynthesis and respiration were evaluated with a differential respirometer. Artemisinin (5 µM) inhibitedL. minor frond production and dry weight 82 and 83%, relative to methanol controls. Chlorophyll content was reduced 44% by artemisinin (2.5 µM). Arteannuic acid (10 µM) was less active, inhibiting frond production 61% and reducing chlorophyll content 66% at 5 µM. Artemisinin (1 µM) reducedL. minor photosynthesis 30% and 2.5 µM reduced respiration 39%. Arteannuic acid had no significant effect on photosynthesis or respiration at the levels tested.
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  • 84
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    Antonie van Leeuwenhoek 66 (1994), S. 151-164 
    ISSN: 1572-9699
    Keywords: purple non-sulfur bacteria ; Rhodobacter ; photosynthesis ; CO2 fixation ; anaerobic respiration ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Purple non-sulfur phototrophic bacteria, exemplifed byRhodobacter capsulatus andRhodobacter sphaeroides, exhibit a remarkable versatility in their anaerobic metabolism. In these bacteria the photosynthetic apparatus, enzymes involved in CO2 fixation and pathways of anaerobic respiration are all induced upon a reduction in oxygen tension. Recently, there have been significant advances in the understanding of molecular properties of the photosynthetic apparatus and the control of the expression of genes involved in photosynthesis and CO2 fixation. In addition, anaerobic respiratory pathways have been characterised and their interaction with photosynthetic electron transport has been described. This review will survey these advances and will discuss the ways in which photosynthetic electron transport and oxidation-reduction processes are integrated during photoautotrophic and photoheterotrophic growth.
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  • 85
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    Antonie van Leeuwenhoek 65 (1994), S. 311-329 
    ISSN: 1572-9699
    Keywords: photosynthesis ; chlorophyll ; bacteriochlorophyll ; reaction center ; electron transfer ; evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Photosynthetic reaction centers from a variety of organisms have been isolated and characterized. The groups of prokaryotic photosynthetic organisms include the purple bacteria, the filamentous green bacteria, the green sulfur bacteria and the heliobacteria as anoxygenic representatives as well as the cyanobacteria and prochlorophytes as oxygenic representatives. This review focuses on structural and functional comparisons of the various groups of photosynthetic reaction centers and considers possible evolutionary scenarios to explain the diversity of existing photosynthetic organisms.
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  • 86
    ISSN: 1573-5060
    Keywords: maize ; Zea mays ; photoinhibition ; photosynthesis ; low-temperature adaptation ; breeding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Sixty-seven inbred lines of maize were evaluated for resistance to low-temperature photoinhibition of photosynthesis, using a pulse-modulated chlorophyll fluorescence technique. The evaluation procedure was based on leaf discs, which were exposed to a high irradiance (1000 µmol/m2/s) at 7°C. The efficiency of open PSII reaction centres as a reflection of overall photosynthesis was measured before and after a photoinhibition-inducing treatment. Exposure of leaf discs to photoinhibitory condition for 2, 4, and 8 hours resulted in an efficiency reduction of 30, 53 and 83%, respectively. Testing of inbred lines showed large differences for photoinhibition susceptibility. The difference in photosynthetic efficiency between the most extreme lines after a treatment of eight hours was 39%. Resistance to photoinhibition was shown to be relevant under cool field conditions. It proved to be a trait strongly amenable to selection.
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  • 87
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    Photosynthesis research 39 (1994), S. 401-412 
    ISSN: 1573-5079
    Keywords: photosynthesis ; elevated CO2 ; source-sink interactions ; Rubisco
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A range of studies of C3 plants have shown that there is a change in both the carbon flux and the pattern of nitrogen allocation when plants are grown under enhanced CO2. This paper examines evidence that allocation of nitrogen both to and within the photosynthetic system is optimised with respect to the carbon flux. A model is developed which predicts the optimal relative allocation of nitrogen to key enzymes of the photosynthetic system as a function of CO2 concentration. It is shown that evidence from flux control analysis is broadly consistent with this model, although at high nitrogen and under certain conditions at low nitrogen experimental data are not consistent with the model. Acclimation to enhanced CO2 is also assessed in terms of resource allocation between photosynthate sources and sinks. A means of assessing the optimisation of this source-sink allocation is proposed, and several studies are examined within this framework. It is concluded that C3 plants probably possess the genetic feedback mechanisms required to efficiently ‘smooth out” any imbalance within the photosynthetic system caused by a rise in atmospheric CO2.
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  • 88
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    Photosynthesis research 39 (1994), S. 453-462 
    ISSN: 1573-5079
    Keywords: photosynthesis ; Rubisco ; ozone ; oxidative stress ; rbcS mRNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Ozone induces reductions in net photosynthesis in a large number of plant species. A primary mechanism by which photosynthesis is reduced is through impact on carbon dioxide fixation. Ozone induces loss in Rubisco activity associated with loss in concentration of the protein. Evidence is presented that ozone may induce oxidative modification of Rubisco leading to subsequent proteolysis. In addition, plants exposed to ozone sustain reduction in rbcS, the mRNA for the small subunit of Rubisco. This loss in rbcS mRNA may lead to a reduced potential for synthesis of the protein. The regulation of O3-induced loss of Rubisco, and implications of the decline in this protein in relation to accelerated senescence are discussed.
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  • 89
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    Photosynthesis research 41 (1994), S. 3-5 
    ISSN: 1573-5079
    Keywords: bacteriochlorophylla-protein ; green sulfur bacteria ; Prosthecochloris aestuarii ; Chlorobium limicola ; photosynthesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In 1961 the green sulfur bacterium-containing mixed culture known as‘Chloropseudomonas ethylicum’ was brought to Brookhaven National Laboratory (USA) from Moscow State University (USSR). The water-soluble bacteriochlorophylla-protein (FMO-protein) was extracted, purified and characterized by absorption and circular dichroism spectroscopy, by X-ray crystallography and by primary structure determination.
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  • 90
    ISSN: 1573-5079
    Keywords: 9-aminoacridine fluorescence ; cyclic electron transport ; Mehler reaction ; photosynthesis ; photosystems ; transthylakoid proton gradient
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The light-dependent quenching of 9-aminoacridine fluorescence was used to monitor the state of the transthylakoid proton gradient in illuminated intact chloroplasts in the presence or absence of external electron acceptors. The absence of appreciable light-dependent fluorescence quenching under anaerobic conditions indicated inhibition of coupled electron transport in the absence of external electron acceptors. Oxygen relieved this inhibition. However, when DCMU inhibited excessive reduction of the plastoquinone pool in the absence of oxygen, coupled cyclic electron transport supported the formation of a transthylakoid proton gradient even under anaerobiosis. This proton gradient collapsed in the presence of oxygen. Under aerobic conditions, and when KCN inhibited ribulose bisphosphate carboxylase and ascorbate peroxidase, fluorescence quenching indicated the formation of a transthylakoid proton gradient which was larger with oxygen in the Mehler reaction as electron acceptor than with methylviologen at similar rates of linear electron transport. Apparently, cyclic electron transport occured simultaneously with linear electron transport, when oxygen was available as electron acceptor, but not when methylviologen accepted electrons from Photosystem I. The ratio of cyclic to linear electron transport could be increased by low concentrations of DCMU. This shows that even under aerobic conditions cyclic electron transport is limited in isolated intact chloroplasts by excessive reduction of electron carriers. In fact, P700 in the reaction center of Photosystem I remained reduced in illuminated isolated chloroplasts under conditions which resulted in extensive oxidation of P700 in leaves. This shows that regulation of Photosystem II activity is less effective in isolated chloroplasts than in leaves. Assuming that a Q-cycle supports a H+/e ratio of 3 during slow linear electron transport, vectorial proton transport coupled to Photosystem I-dependent cyclic electron flow could be calculated. The highest calculated rate of Photosystem I-dependent proton transport, which was not yet light-saturated, was 330 μmol protons (mg chlorophyll h)−1 in intact chloroplasts. If H+/e is not three but two proton transfer is not 330 but 220 μmol (mg Chl H)−1. Differences in the regulation of cyclic electron transport in isolated chloroplasts and in leaves are discussed.
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  • 91
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    Photosynthesis research 40 (1994), S. 35-44 
    ISSN: 1573-5079
    Keywords: allophycocyanin ; chlorophyll a ; linear dichroism ; phycobilisome ; photosynthesis ; state transition
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Low temperature (77 K) linear dichroism spectroscopy was used to characterize pigment orientation changes accompanying the light state transition in the cyanobacterium, Synechococcus sp. PCC 6301 and those accompanying chromatic acclimation in Porphyridium cruentum in samples stabilized by glutaraldehyde fixation. In light state 2 compared to light state 1 intact cells of Synechococcus showed an increased alignment of allophycocyanin parallel to the cells' long axis whereas the phycobilisomethylakoid membrane fragments exhibited an increased allophycocyanin alignment parallel to the membrane plane. The phycobilisome-thylakoid membrane fragments showed less alignment of a short wave-length chlorophyll a (Chl a) Qy transition dipole parallel to the membrane plane in state 2 relative to state 1. To aid identification of the observed Chl a orientation changes in Synechococcus, linear dichroism spectra were obtained from phycobilisome-thylakoid membrane fragments isolated from red light-grown (increased number of PS II centres) and green light-grown (increased number of PS I centres) cells of the red alga Porphyridium cruentum. An increased contribution of short wavelength Chl a Qy transition dipoles parallel to the long axis of the membrane plane was directly correlated with increased levels of PS II centres in red light-grown P. cruentum. Our results indicate that the transition to state 2 in cyanobacteria is accompanied by an increase in the orientation of allophycocyanin and a decrease in the orientation of Chl a associated with PS II with respect to the thylakoid membrane plane.
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  • 92
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    Photosynthesis research 40 (1994), S. 75-92 
    ISSN: 1573-5079
    Keywords: chilling temperature ; oxygen toxicity ; photoinhibition ; photosynthesis ; photosystems ; superoxide dismutase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract When 23 °C-grown potato leaves (Solanum tuberosum L.) were irradiated at 23 °C with a strong white light, photosynthetic electron transport and Photosystem-II (PS II) activity were inhibited in parallel. When the light treatment was given at a low temperature of 3 °C, the photoinhibition of photosynthesis was considerably enhanced, as expected. Surprisingly, no such stimulation of photoinhibition was observed with respect to the PS II function. A detailed functional analysis of the photosynthetic apparatus, using in-vivo fluorescence, absorbance, oxygen and photoacoustic measurements, and artificial electron donors/acceptors, showed a pronounced alteration of PS I activity during light stress at low temperature. More precisely, it was observed that both the pool of photooxidizeable reaction center pigment (P700) of PS I and the efficiency of PS I to oxidize P700 were dramatically reduced. Loss of P700 activity was shown to be essentially dependent on atmospheric O2 and to require a continued flow of electrons from PS II, suggesting the involvement of the superoxide anion radical which is produced by the interaction of O2 and the photosynthetic electron-transfer chain through the Mehler reaction. Mass spectrometric measurements of O2 exchange by potato leaves under strong illumination did not reveal, however, any stimulation of the Mehler reaction at low temperature, thus leading to the conclusion that O2 toxicity mainly resulted from a chilling-induced inhibition of the scavenging system for O2-radicals. Support for this interpretation was provided by the light response of potato leaves infiltrated with an inhibitor (diethyldithiocarbamate) of the chloroplastic Cu-Zn superoxide dismutase. It was indeed possible to simulate the differential inhibition of the PS II photochemical activity and the linear electron transport observed during light stress at low temperature by illuminating at 23 °C diethyldithiocarbamate-poisoned leaves. The experimental data presented here suggests that (i) the previously reported resistance of PS I to photoinhibition damage in-vivo is not an intrinsic property of PS I but results from efficient protective systems against O2 toxicity, (ii) PS I is photoinhibited in chilled potato leaf due to the inactivation of this PS I defence system and (iii) PS I is more sensitive to superoxide anion radicals than PS II.
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  • 93
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    Photosynthesis research 40 (1994), S. 207-221 
    ISSN: 1573-5079
    Keywords: photosynthesis ; unicellular algae ; ion effects ; manganese ; blue light ; synchronized growth ; algal mass cultures
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract This personal perspective records research experiences in chemistry and biology at four German universities, two before and two after World War II. The research themes came from cytophysiology of green unicellular algae, in particular their photosynthesis. The function of inorganic ions in photosynthesis and dark respiration was investigated at different degrees of specific mineral stress (deficiencies), and the kinetics of recovery followed after the addition of the missing element. Two types of recovery of photosynthesis were observed: indirect restitution via growth processes and immediate normalisation. From the latter case (K+, phosphate, Mn++) the effect of manganese was emphasized as its role in photosynthetic O2 evolution became established during our research. Other themes of our group, with some bearing on photosynthesis were: synchronization of cell growth by light-dark change and effects of blue (vs. red) light on the composition of green cells. Some experiences in connection with algal mass cultures are included. Discussion of several editorial projects shows how photosynthesis, as an orginally separated field of plant biochemistry and biophysics, became included into general cell physiology and even ecophysiology of green plants. The paper contains an appreciation of the authors' main mentor Kurt Noack (1888–1963) and of Ernst Georg Pringsheim (1881–1970), founder of experimental phycology.
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  • 94
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    Photosynthesis research 41 (1994), S. 27-28 
    ISSN: 1573-5079
    Keywords: photosynthesis ; reaction center ; green bacteria ; heliobacteria ; gene ; chlorosome ; cytochrome
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Recommendations are given for naming of genes coding for reaction center, antenna and electron transport proteins in green photosynthetic bacteria and heliobacteria
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  • 95
    ISSN: 1573-5079
    Keywords: photosynthesis ; Chlorobium tepidum ; antenna ; bacteriochlorophylla protein ; energy transfer ; chlorosome ; green bacteria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The BChla-containing Fenna-Matthews-Olson (FMO) protein from the green sulfur bacteriumChlorobium tepidum was purified and characterized. Fluorescence spectra indicate that efficient excited state quenching occurs at neutral or oxidizing redox potentials. The major fluorescence lifetime at room temperature is approximately 60 ps in samples that are in neutral or oxidizing conditions, and approximately 2 ns in samples where the strong reductant sodium dithionite has been added. A similar change is observed in pump-probe picosecond absorbance difference experiments, where the long life time component increases after dithionite addition. A 16 Gauss wide EPR signal with g factor =2.005 is observed in samples without dithionite. This signal largely disappears upon addition of dithionite. Dithionite induces large reversibile changes in the 77 K absorbance spectra of the purified FMO protein and in whole cells. These results indicate that the FMO protein contains redox active groups, which may be involved in the regulation of energy transfer. Room temperature circular dichroism and low temperature absorption spectra show that dithionite also induces conformational or structural changes of the FMO protein complex.
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  • 96
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    Plant and soil 164 (1994), S. 97-105 
    ISSN: 1573-5036
    Keywords: alcohol dehydrogenase ; drought ; flooding ; photosynthesis ; tropical grasses ; water potential
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Long dry seasons or permanent flooding, typical of tropical savannas, severely limit the growth of pasture plants. This study compares the responses of water relations, carbon assimilation and alcohol dehydrogenase (ADH) activity to drought and flooding in four perennial C4 grasses: the tufted or caespitose Hyparrhenia rufa and Andropogon gayanus (CIAT 621) and the stoloniferous Brachiaria mutica and Echinochloa polystachya. Plants of the four species were subjected to medium term flooding (20–25 days) and moderate drought in a greenhouse. Leaf water potential (Ψ), stomatal conductance (Gs) and photosynthesis rate (Pn) were measured throughout the experiment and ADH activity was measured in flooded and control plants. Moderate drought produced similar effects in all grasses reducing Gs which caused reduced Pn. Net photosynthesis compensation point was reached at the lowest Ψ in A. gayanus which was considered as the most drought tolerant. The responses to flooding varied across species. Andropogon gayanus and H. rufa showed early stomatal closure without concurrent decrease in Ψ and leaf turgor. This low Gs was responsible of reduced Pn and growth rate. There was a slight increase of Gs in the middle of the flooding period and both grasses recovered pre-stress Gs and Pn after drainage. ADH activity increased markedly only in A. gayanus under flooding suggesting that this grass was the most flood-sensitive. Stomatal aperture, Pn and ADH activity in B. mutica and E. polystachya were not affected by flooding. The higher flood-tolerance in these grasses might be attributed to enhanced oxygen diffusion to the roots through the hollow stolons, development of advantitious rootlets and large aerenchyma in the roots which compensate for the reduction of soil oxygen and permit the maintenance of root activity.
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  • 97
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    Euphytica 76 (1994), S. 235-238 
    ISSN: 1573-5060
    Keywords: Triticum aestivum ; wheat ; photosynthesis ; gene action ; heritability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Gene action and heritability for photosynthetic activity were estimated from generation means in two wheat crosses during two stages (5 th leaf and flag leaf between 2 and 5 days after anthesis). Six generations were available for each cross: parents (P1 and P2), F1, F2 and backcrosses (BC1 and BC2). Correlations between some morphophysiological characters and photosynthetic activity of the flag leaf was also determined. The joint scaling test described by Mather & Jinks was used to determine the gene action. It showed that them; [d]; [h]; [i], [l] (mean, additivity, dominance, additive x additive interallelic interaction effects, dominance x dominance interallelic interaction effects) model fits the two crosses at both measurement times. All the model genetic components were significant for the flag leaf, however for the 5 th leaf only [h]; [i] and [l] were significant. The presence of additive and additive x additive effects suggested the possibility of selecting for this character using the flag leaf so as to obtain pure inbred lines. Dominance effects [h] were negative and dominance x dominance effects [l] were positive. Broad sense heritability values were medium to low. There were no correlations between the studied morphophysiological characters and the photosynthetic activity.
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  • 98
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    Photosynthesis research 39 (1994), S. 191-199 
    ISSN: 1573-5079
    Keywords: photoinhibition ; photosynthesis ; protein degradation ; singlet oxygen ; TEMP (2,2,6,6-tetramethylpiperidine) ; thylakoid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Exposure of isolated spinach thylakoids to high intensity illumination (photoinhibition) results in the well-characterized impairment of Photosystem II electron transport, followed by degradation of the D1 reaction centre protein. In the present study we demonstrate that this process is accompanied by singlet oxygen production. Singlet oxygen was detected by EPR spectroscopy, following the formation of stable nitroxide radicals from the trapping of singlet oxygen with a sterically hindered amine TEMP (2,2,6,6-tetramethylpiperidine). There was no detectable singlet oxygen production during anaerob photoinhibition or in the presence of sodium-azide. Comparing the kinetics of the loss of PS II function and D1 protein with that of singlet oxygen trapping suggests that singlet oxygen itself or its radical product initiates the degradation of D1.
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  • 99
    ISSN: 1573-5079
    Keywords: cyanobacteria ; excitation energy ; herbicide ; photochemical apparatus ; photosynthesis ; pyridazinone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract SANDOZ 9785, also known as BASF 13.338, is a pyridazinone derivative that inhibits Photosystem II (PS II) activity leading to an imbalance in the rate of electron transport through the photosystems. Synechococcus sp. strain PCC 7942 cells grown in the presence of sublethal concentration of SANDOZ 9785 (SAN 9785) for 48 hours exhibited a 20% decrease in Chl a per cell. However, no changes were observed in the content of phycocyanin per cell, the size of the phycobilisomes or in the PS II:PS I ratio. From an estimate of PS II electron transport rate under varying light intensities and spectral qualities and analysis of room temperature Chl a fluorescence induction, it was deduced that growth of Synechococcus PCC 7942 in the presence of SAN 9785 leads to a redistribution of excitation energy in favour of PS II. Though the redistribution appears to be primarily caused by changes affecting the Chl a antenna of PS II, the extent of energetic coupling between phycobilisomes and PS II is also enhanced in SAN 9785 grown Synechococcus PCC 7942 cells. There was a reduction in the effective size of PS I antenna based on measurement of P700 photooxidation kinetics. These results indicate that when PS II is partially inhibited, the structure of photosynthetic apparatus alters to redistribute the excitation energy in favour of PS II so that the efficiency of utilization of light energy by the two photosystems is optimized. Our results suggest that under the conditions used, drastic structural changes are not essential for redistribution of excitation energy between the photosystems.
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    Photosynthesis research 39 (1994), S. 351-368 
    ISSN: 1573-5079
    Keywords: CO2 enrichment ; gas exchange ; greenhouse effect ; photosynthesis ; responses to CO2
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The nature of photosynthetic acclimation to elevated CO2 is evaluated from the results of over 40 studies focusing on the effect of long-term CO2 enrichment on the short-term response of photosynthesis to intercellular CO2 (the A/Ci response). The effect of CO2 enrichment on the A/Ci response was dependent on growth conditions, with plants grown in small pots (〈 5 L) or low nutrients usually exhibiting a reduction of A at a given Ci, while plants grown without nutrient deficiency in large pots or in the field tended to exhibit either little reduction or an enhancement of A at a given Ci following a doubling or tripling of atmospheric CO2 during growth. Using theoretical interpretations of A/Ci curves to assess acclimation, it was found that when pot size or nutrient deficiency was not a factor, changes in the shape of A/Ci curves which are indicative of a reallocation of resources within the photosynthetic apparatus typically were not observed. Long-term CO2 enrichment usually had little effect or increased the value of A at all Ci. However, a minority of species grown at elevated CO2 exhibited gas exchange responses indicative of a reduced amount of Rubisco and an enhanced capacity to metabolize photosynthetic products. This type of response was considered beneficial because it enhanced both photosynthetic capacity at high CO2 and reduced resource investment in excessive Rubisco capacity. The ratio of intercellular to ambient CO2 (the Ci/Ca ratio) was used to evaluate stomatal acclimation. Except under water and humidity stress, Ci/Ca exhibited no consistent change in a variety of C3 species, indicating no stomatal acclimation. Under drought or humidity stress, Ci/Ca declined in high-CO2 grown plants, indicating stomata will become more conservative during stress episodes in future high CO2 environments.
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