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  • Articles  (713)
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  • 1
    ISSN: 1432-1211
    Keywords: Key words MHC class III region ; Mouse ; Human ; G7c ; Lung tumor susceptibility
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Immunogenetics 51 (2000), S. 487-488 
    ISSN: 1432-1211
    Keywords: Key words Immunoglobulin ; J segments ; IGKJ genes ; Alleles ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
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  • 3
    ISSN: 1573-4919
    Keywords: lipids ; nucleus ; liver ; sex ; bezafibrate ; rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The aim of the present study was to investigate whether lipid metabolism in the nuclei is affected by changes in the metabolism of free fatty acids in the liver. The experiments were carried out on 3 groups of rats: 1 - control-male, 2 - female, and 3 - male, treated with bezafibrate (a peroxisome proliferator). The rats received 14C-palmitic acid intravenously. Thirty min later liver samples and blood from the abdominal aorta were taken. The liver nuclei were isolated in sucrose gradient. Lipids were extracted from the nuclei and the liver homogenate and subsequently separated into the following fractions: phospholipids, mono-, di- and triacylglycerols, free fatty acids, cholesterol and cholesterol esters. The radioactivity of each fraction was counted. Furthermore, the content of free fatty acids and the fatty acid binding proteins was measured. It was found that radioactivity was present in each lipid fraction obtained from the liver homogenate and from the nuclei. In the female group, the total radioactivity of lipids in the liver homogenate was lower, whereas in the nuclei it was higher in comparison to the male group. The reduction in the radioactivity in the liver was mostly accounted for by decreased radioactivity in the fraction of triacylglycerols and phospholipids. In the nuclei, the radioactivity of the fraction of phospholipids, free fatty acids and diacylglycerols was elevated. Bezafibrate did not affect the total radioactivity of lipids in the liver and reduced it in the nuclei. In the liver, the drug increased radioactivity mostly in the fraction of phospholipids and reduced it mainly in the fraction of triacylglycerols. In the nuclei, the radioactivity of each lipid fraction examined was reduced. The content of the fraction of free fatty acids in the liver and in the nuclei in the female and in the bezafibrate-treated groups did not differ from the respective value in the control group. The content of fatty acid binding proteins in the nuclei of the female and bezafibrate-treated groups increased in parallel to the elevation in their content in the cytosol. It is concluded that the female sex hormones and bezafibrate influence the transport of selected lipids into the nuclei. The effects seem to be a consequence of the action of these factors directly on the nucleus.
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  • 4
    ISSN: 1573-4919
    Keywords: liver ; nucleic-acid concentrations ; Oncorhynchus mykiss ; protein-growth rate ; protein-turnover rate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract We have determined the protein-turnover rates and nucleic-acid concentrations in the liver of trout (Oncorhynchus mykiss) fed on two different isocaloric diets: low-protein/high-fat and non-carbohydrate/high-fat. Compared to controls, the partial replacement of protein with fat significantly decreased the protein accumulation rate and protein-retention efficiency in the liver whilst increasing the fractional protein-synthesis and protein-degradation rates as well as protein-synthesis efficiency. The complete replacement of carbohydrates with fat significantly lowered the protein-accumulation rate and protein-retention efficiency, but enhanced both the protein-synthesis and protein-degradation rates as well as protein-synthesis capacity. The protein:DNA and RNA:DNA ratios decreased considerably on both diets. Total DNA decreased in fish on a low-protein/high-fat diet but did not change in those on a non-carbohydrate/high-fat diet. The absolute protein-synthesis rate registered no significant change under any of the nutritional conditions. Both the experimental diets did however raise the fractional protein-synthesis rate significantly, due to enhanced protein-synthesis efficiency when protein was partially replaced with fat and to enhanced protein-synthesis capacity when carbohydrates were completely replaced with fat. Our results show the capacity of the liver to adapt its turnover rates and conform to different nutritional conditions. They also point to the possibility of controlling fish growth by dietary means.
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Molecular and cellular biochemistry 207 (2000), S. 101-104 
    ISSN: 1573-4919
    Keywords: insulin-like growth factor binding protein ; liver ; cancer ; hepatocellular carcinoma ; liver disease ; insulin-like growth factor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Insulin-like growth factors (IGF), IGF receptors and IGF binding proteins (IGFBPs) play an important role in cell growth and differentiation. The liver is the major source of IGF-1 and at least two IGFBPs (IGFBP-1 and IGFBP-3). IGFBPs most often serve to attenuate the effects of IGF at the receptor level and thereby limit IGF-induced cell growth and differentiation. Although changes in IGFBP expression have been described during controlled liver growth such as hepatic regeneration following partial hepatectomy, there is limited knowledge of IGFBPs gene expression in uncontrolled growth or hepatocellular carcinoma. In the present study, we employed Northern blotting techniques to document the expression of IGFBP-1, 3 and 4 in normal human livers, cirrhotic and hepatocellular carcinoma tissues. The results revealed no differences in IGFBP-1, 3 and 4 mRNA levels between normal and cirrhotic tissues. However, the expression of all three IGFBPs mRNA were significantly down regulated in hepatocellular carcinoma tissues. These findings are in keeping with IGFBPs playing an important inhibitory role in the development and/or growth of hepatocellular carcinoma in humans.
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Water resources management 14 (2000), S. 157-175 
    ISSN: 1573-1650
    Keywords: estuarine management ; fishery harvest ; freshwater inflow ; linear quadratic regulator ; salinity ; successive approximation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Architecture, Civil Engineering, Surveying , Geography
    Notes: Abstract A successive approximation linear quadratic regulator (SALQR)method is applied to solve estuarine management problems to determine the optimal amount of freshwater inflows into baysand estuaries to maximize fishery harvests. Fishery harvests areexpressed in regression equations as functions of freshwaterinflows. The optimization problem is posed as a discrete-timeoptimal control problem in which salinity represents the statevariable and freshwater inflow represents the control variable. A two-dimensional hydrodynamic-salinity transport model, HYD-SAL,is used as the transition to simulate the flow circulation andtemporal and spatial salinity pattern in an estuary system. Thebound constraints for the control and state variables areincorporated into the objective function using a penalty functionmethod to convert the problem into an unconstrained formulation. The SALQR method is applied to the Lavaca-Tres Palacios Estuaryin Texas and the results are compared with those of usingregression equations as the transition equations.
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  • 7
    ISSN: 1573-675X
    Keywords: false-positive ; immunocytochemistry ; kidney ; liver ; mouse ; TUNEL
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Apoptosis is a physiological, programmed process for the elimination of cells from living organisms. Currently, one of the most frequently used methods to detect apoptosis is TUNEL assay. It has provided valuable information about apoptosis in various tissues. However, the sensitivity and the specificity of TUNEL technique have also been criticized. We detected an intense false-positive apoptotic signal in nude and Balb/c mice kidney and liver. In kidney the signal was confined to the proximal, distal and collecting tubular cells, and in liver to hepatocytes. Both tissues appeared normal in light microscopy, and no DNA ladder formation or increase in caspase-3 enzyme activity was detected. BrdU labelling and Ki-67 immunostaining did not reveal increased cell proliferation in these tissues. On the other hand, false-positive signal was not detected in testis, spleen, pancreas or renal cell carcinoma from the same animals. Also, no false-positive signal was seen in human liver or kidney samples. Although factors known to produce false-positive staining related to sample harvesting, preparation and staining protocols were eliminated, the cause of the false- positive apoptotic signal remains unknown. We conclude that caution must be exercised when examining apoptosis in mouse tissues with TUNEL assay.
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Bulletin of experimental biology and medicine 130 (2000), S. 1001-1004 
    ISSN: 1573-8221
    Keywords: thymus ; liver ; human fetus ; thymaline
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The endocrine function of the thymus develops earlier than lymphocytopoietic. Thymalin is produced by epithelial cells in the thymus primordium. It is released into the blood and regulates differentiation of T lymphocytes in the liver, the initial hemopoietic organ. The hormonal and lymphopoietic functions of human thymus are united on weeks 7.5-8 of embryonic life.
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  • 9
    ISSN: 1573-8221
    Keywords: Reye's syndrome ; 4-pentenoic acid ; eplir ; essentiale ; liver
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Hepatoprotectors containing phospholipids exert a therapeutic effect in experimental Reye's syndrome in rats induced by 40pentenoic acid. Eplir more effectively than essentiale decreased necrosis of the liver, reduced serum enzymes and bilirubin, alleviated hypoglycemia, suppressed MDA production, and improved ketogenesis and detoxication of ammonium and phenols.
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Bulletin of experimental biology and medicine 129 (2000), S. 605-607 
    ISSN: 1573-8221
    Keywords: Kupffer cell ; tetrachloromethane ; liver
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Acute toxic hepatitis was modeled in (CBA×C57B1)F1 mice by single injection of 40% CCl4 in oil. Pretreatment with gadolinium chloride, a selective blocker of Kupffer cells, considerably potentiated damage to hepatocytes leading to generalization of this process, delayed inflammatory infiltration, and inhibited reparative processes. Zymosan administered against the background of gadolinium chloride blockade improved liver resistance to CCl4-induced damage, intensified mononuclear infiltration, and accelerated reparative processes.
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  • 11
    Electronic Resource
    Electronic Resource
    Springer
    Bulletin of experimental biology and medicine 129 (2000), S. 350-351 
    ISSN: 1573-8221
    Keywords: hyperthermia ; liver ; perfusion ; metabolism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The metabolic state of rat liver was studiedin vivo 1, 6, and 18 h after single hyperthermia (42°C). It was shown that intracellular compensatory reactions involved in restoration of the energy state and realization of endogenous cytoprotective mechanisms play a role during recovery.
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  • 12
    Electronic Resource
    Electronic Resource
    Springer
    Bulletin of experimental biology and medicine 130 (2000), S. 1209-1211 
    ISSN: 1573-8221
    Keywords: liver ; sinusoidal cells ; endotheliocytes ; Kupffer cells ; lipocytes ; 3,4-benzopyrene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Reorganization of sinusoidal cells (endotheliocytes, Kupffer cells, and Ito cells) in the liver microregion of male rat liver exposed to 3,4-benzopyrene was studied. Synchronous activation of the lysosome-vacuole systems in Kupffer cells and endotheliocyte indicates cooperation of these cells in detoxification of benzopyrene and its metabolites. Ito cells lose lipid inclusions and actively proliferate; the appearance of intermediate forms between lipocytes and fibroblasts attests to activation of fibrogenesis in the liver.
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  • 13
    Electronic Resource
    Electronic Resource
    Springer
    Bulletin of experimental biology and medicine 129 (2000), S. 524-526 
    ISSN: 1573-8221
    Keywords: macrophages ; Kupffer cells ; liver ; CCl 4
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Acute toxic hepatitis in male Wistar rats was produced by single injection of 40% CCl4 (0.2 ml per 100 g body weight in oil). Pretreatment with various immunostimulators (bacterial polysaccharides prodigiozan and salmozan; yeast polysaccharides zymosan, peptidoglycan, and mannan; and hydrolytic enzyme egg lysozyme) produced a hepatoprotective effect correlating which the stimulatory influence on macrophages and increasing in the following order: mannan〈peptidoglycan〈zymosan〈lysozyme〈salmozan〈prodigiozan.
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  • 14
    Electronic Resource
    Electronic Resource
    Springer
    Medical & biological engineering & computing 38 (2000), S. 454-464 
    ISSN: 1741-0444
    Keywords: Optic nerve ; Human ; Self-sizing spiral cuff electrode ; Finite elements method ; Fibre model
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The aim of the study is to determine which of the existing myelinated mammalian nerve fibre models better fits experimental data resulting from electrical stimulation of the human optic nerve and from propagation velocity measured on primates. The macroscopic electric potential is computed in a 3D, inhomogeneous and anisotropic nerve model. The Chiu-Sweeney (CS) and the Schwarz-Wesselink (SW) membrane descriptions are then considered. Variations in parameters that are not well established (encapsulation-tissue thickness, nerve-fascicle conductivity, geometric and electrochemical fibre cable parameters) are taken into account. Results demonstrate that the SW model predictions are in better agreement with the experimental data than those of the CS model, although thresholds are still too high. When channel densities are varied, the SW model turns out to be more robust than the CS model. For a suitable leakage channel density value (about 8% of the original one), the SW model predicts a conduction velocity of 11.4ms−1 and an excitation threshold of 0.055 mA (for 0.1 ms pulse duration), which is in very good agreement with experimental values (11 ms−1 and 0.055 mA). Potassium current in the SW model is necessary for stability. Introduction of a potassium-like current can restore stability in the CS system.
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  • 15
    Electronic Resource
    Electronic Resource
    Springer
    Journal of comparative physiology 185 (1999), S. 297-304 
    ISSN: 1432-1351
    Keywords: Key words Odor coding ; Learning ; Enhanced sensitivity ; Rabbit ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The olfactory system is faced with a particular problem – the high dimensionality and inherent unpredictability of the chemical world. Most natural odorants encountered in everyday life are complex mixtures of many different volatiles. This means that from the outset the olfactory system has to contend with a great and often unpredictable diversity of molecules, making it difficult for stable primary features of the chemical world to be mapped onto the sensory surface. One solution to such unpredictability is provided by learning. Learning confers flexibility, enabling individuals of a given species to acquire and make use of the most appropriate information in a particular environment. Two examples of this are presented: learning of maternal odors in neonatal rabbits, including evidence that the sensory surface itself may be influenced by environmental conditions so as to enhance sensitivity to molecules of particular ecological relevance, and cross-cultural human studies suggesting that experience with everyday odors influences not only the way these are evaluated, but also their perceived intensity. It is concluded that an adequate understanding of odor coding and olfactory function will not be possible without taking such experience-dependent factors into account.
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  • 16
    ISSN: 1573-0832
    Keywords: interspecific interactions ; ochratoxin ; Aspergillus ochraceus ; spoilage fungi ; water relations ; temperature
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The effect of water availability (water activity,aw; 0.995–0.90) and temperature (18–30 °) on in vitro interactions between an ochratoxin producing strain of Aspergillus ochraceus and six other spoilage fungi was assessed in dual culture experiments on a maize meal-based agar medium. Inprimary resource capture of nutrient substrate, A. ochraceus was dominant against many of the interacting species, being able to overgrow and replace A. candidus, and sometimes A. flavus and the Eurotium spp. regardless of aw or temperature. However, with freely available water (0.995 aw) A. alternata and A. niger were dominant, with mutual antagonism between A. ochraceus and A. flavus at 25–30 °C. In the driest conditions tested (0.90 aw) there was also mutual antagonism between A. ochraceus and the two Eurotium spp. Overall, under allconditions tested the Index of Dominance for A. ochraceus was much higher than for other competing species combined suggesting that A. ochraceus wasa good competitive colonist able to replace a numberof other species. However, the growth rate ofA. ochraceus was modified and decreased by the interaction with competitors. Interaction between A. ochraceus and species such as A. alternata (18°C/0.995) and Eurotium spp. (0.995–0.95 and 25–30 °C) resulted in a significant stimulation of ochratoxin production. Theresults are discussed in relation to the effect that environmental factors have on the possible competitiveness of A. ochraceus in the maizegrain ecosystem and the role of ochratoxin in nicheexclusion of competitors.
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  • 17
    Electronic Resource
    Electronic Resource
    Springer
    Immunogenetics 49 (1999), S. 438-445 
    ISSN: 1432-1211
    Keywords: Key words PA28 ; Proteasome ; Gene structure ; Evolution ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Two proteasome activators PA28α and β, which have been implicated in antigen processing for loading class I MHC molecules, are synthesized in response to Ifn-γ. The human genes encoding these activators (PSME1 and PSME2, respectively) were analyzed by sequencing. Each gene comprised 11 exons, consistent with gene duplication during vertebrate evolution. The intron/exon organization of both genes was highly conserved, the major difference being the absence of the exon encoding the lysine and glutamic acid-rich 'KEKE' motif in PA28β. Two other genes of relevance to the immune system were located close to those for PA28 at 14q11.2 including ISGF3G, a protein involved in transcription after IFNα signalling. These sequences were also characterized.
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  • 18
    ISSN: 1432-1211
    Keywords: Key words NK cells ; Human ; Surface molecule ; Lectin superfamily ; NK gene complex
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Natural killer (NK) cells constitute the third major population of lymphocytes. They possess the inherent capacity to kill various tumor and virally infected cells and mediate the rejection of bone-marrow grafts in lethally irradiated animals. A large family of NK cell receptors belong to the C-type lectin superfamily and are localized to the NK gene complex on Chromosome (Chr) 6 in the mouse and Chr 12 in the human. Genes in the NK gene complex encode type II receptors and examples include the families of NKR-P1, Ly-49, and NKG2 receptors. Examples of other C-type lectin-like NK cell receptors that occur as individual genes are CD94, CD69, and AICL. Here we report the molecular characterization and chromosomal mapping of a human lectin-like transcript (LLT1) expressed on NK, T, and B cells and localized to the NK gene complex within 100 kilobases of CD69. The cDNA encodes a predicted protein of 191 amino acid residues with a transmembrane domain near the N-terminus and an extracellular domain of 132 amino acid residues with similarity to the carbohydrate recognition domain of C-type lectins. The predicted protein of LLT1 shows 59 and 56% similarity to AICL and CD69, respectively. The predicted protein does not contain any intracellular ITIM motifs, suggesting that LLT1 may be involved in mediating activation signals.
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  • 19
    Electronic Resource
    Electronic Resource
    Springer
    European journal of clinical pharmacology 55 (1999), S. 349-352 
    ISSN: 1432-1041
    Keywords: Key words Ischaemic stroke ; Vinpocetine ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Abstract Objectives: To determine whether vinpocetine decreases short- and long-term case fatality and proportion of dependent survivors if administered within 2 weeks of stroke onset. Methods: All published and unpublished trials were attempted to be identified using the standard search strategy of the Cochrane Collaboration Stroke Review Group, using MEDLINE searches performed with all known manufacturer code names and trade names of vinpocetine and by contacting manufacturers of vinpocetine to give information of all randomised controlled trials on vinpocetine in stroke. Researchers who participated in trials on vinpocetine in Hungary were asked for further information. Only truly randomised, unconfounded clinical trials that compared the effect of vinpocetine to either placebo or another reference treatment for acute stroke where treatment started no later than 14 days after stroke onset were eligible for inclusion. Data synthesis and analysis was performed using the Cochrane Review Manager software (RevMan version 3.0). Results: Among the identified studies on vinpocetine in stroke, only one fulfilled the selection criteria for inclusion in the review. No death occurred in the study groups and no statistically significant difference was found in dependency between the treatment and the placebo groups. No adverse effects were reported. Conclusions: Based on only one small randomised controlled unconfounded study, presently there is not enough evidence to decide whether the administration of vinpocetine does or does not decrease case fatality and dependency in acute stroke.
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  • 20
    ISSN: 1573-4919
    Keywords: Bcl-2 ; Ca2+ ; hepatoma ; liver ; mitochondrial permeability transition ; tumor cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Zajdela hepatoma mitochondria were able to accumulate two to five times more Ca2+ than rat liver mitochondria before the permeability transition was induced. Pulses of Ca2+ were given in series to determine the Ca2+ threshold by recording changes in [Ca2+] and membrane potential, the permeability transition causing the release of accumulated Ca2+ and collapse of the membrane potential. Hepatoma mitochondria had lower Ca2+ efflux rates, higher net Ca2+ uptake rates and lower phosphorylation rates than liver mitochondria. Since the differences in regard to induction of the permeability transition might be due to higher expression of the Bcl-2 protein in hepatoma cells than in hepatocytes, the transcription of Bcl-2 and the proteins reacting with a Bcl-2 polyclonal antiserum were estimated by Northern and Western blotting, respectively. Hepatoma cells had two Bcl-2 specific mRNA bands of 7 and 2.4 kb, and substantial amounts of the Bcl-2 protein, whereas in liver cells and mitochondria these were not detected. Both cell lines had a reactive band at 19-20 kDa, and hepatocytes a small band at 31-32 kDa. Bcl-2 antibodies stimulated the permeability transition potently in hepatoma mitochondria.
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  • 21
    ISSN: 1573-4919
    Keywords: hyperplasia ; hypertrophy ; liver ; nucleic-acid concentration ; protein-growth rate ; protein-turnover rate ; rainbow trout (Oncorhynchus mykiss) ; starvation/re-feeding cycle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract We report upon the effects of a cycle of long-term starvation followed by re-feeding on the liver-protein turnover rates and nature of protein growth in the rainbow trout (Oncorhynchus mykiss). We determined the protein-turnover rate and its relationship with the nucleic-acid concentrations in the livers of juvenile trout starved for 70 days and then re-fed for 9 days. During starvation the total hepatic-protein and RNA contents decreased significantly and the absolute protein-synthesis rate (AS) also fell, whilst the fractional protein-synthesis rate (KS) remained unchanged and the fractional protein-degradation rate (KD) increased significantly. Total DNA content, an indicator of hyperplasia, and the protein:DNA ratio, an indicator of hypertrophy, both fell considerably. After re-feeding for 9 days the protein-accumulation rates (KG, AG) rose sharply, as did KS, AS, KD, protein-synthesis efficiency (KRNA) and the protein-synthesis rate/DNA unit (KDNA). The total hepatic protein and RNA contents increased but still remained below the control values. The protein:DNA and RNA:DNA ratios increased significantly compared to starved fish. These changes demonstrate the high response capacity of the protein-turnover rates in trout liver upon re-feeding after long-term starvation. Upon re-feeding hypertrophic growth increased considerably whilst hyperplasia remained at starvation levels.
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  • 22
    ISSN: 1432-0878
    Keywords: Key words Apoptosis ; p53 ; Ischemia ; Enterocytes ; Proliferation ; Differentiation ; ISEL ; Glomeruli ; Mouse (Balb/c) ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Morphological changes associated with apoptosis are closely correlated with the expression of specific proteins. However, the cause-effect relationships between the expression of these proteins and DNA degradation are barely known. For studying expression of apoptosis-related proteins in relation to different degrees of DNA fragmentation, the small intestine with its spatially organized continuum of proliferation, differentiation and death is a very useful preparation. Enterocytes towards the apex of the villi become increasingly susceptible to apoptosis. Here, this ”apoptotic gradient” is used to demonstrate the presence of BAX and BCL-2 proteins in the cytoplasm of cells at the onset of apoptosis. In semithin serial sections of the small intestine, BAX, BCL-2 and DNA fragmentation were demonstrated. BAX and BCL-2 are always colocalized and only in cells with fragmented DNA. The gradient of BAX or BCL-2 staining is similar to the gradient of DNA fragmentation. Immunoreactivity for BCL-2 or BAX is most intense in cells that are prone to become apoptotic next in the course of cellular turnover but not in cells in an advanced apoptotic state, showing strongly condensed chromatin. When using the same technique on semithin sections of kidney biopsies, containing epithelia with low cellular turnover, we found DNA fragmentation mainly in the epithelial cells of the distal tubules. Similar to the situation in the enterocytes, BAX staining was confined to the cytoplasm of epithelial cells with a moderate degree of DNA fragmentation and reduced in epithelial cells with a high degree of DNA fragmentation. In contrast to the situation in the small intestine, very low levels of BCL-2 were found. The results suggest that expression of BCL-2 and BAX is related to cell damage as indicated by DNA fragmentation but not to advanced stages of cellular death, as indicated by chromatin condensation and cellular shrinkage.
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  • 23
    ISSN: 1432-0878
    Keywords: Key words Vesicle protein ; Physin ; Secretory carrier-associated membrane protein ; Vesicle-associated membrane protein ; Synaptobrevin ; Expression ; Liver ; Bovine ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The cell-type restricted expression of cytoplasmic microvesicle membrane protein isoforms may be a consequence of the functional adaptation of these vesicles to the execution of specialized processes in cells of different specialization. To characterize the expression of the vesicle protein pantophysin, an isoform of the synaptic vesicle proteins synaptophysin and synaptoporin, we have prepared and characterized antibodies useful for the immunological detection of pantophysin in vitro and in situ. Using these reagents, we show by immunoblot analyses that pantophysin expression is not homogeneous but differs significantly between various bovine tissues. Furthermore, these differences are not exactly paralleled by the expression of other vesicle proteins of the SCAMP (secretory carrier-associated membrane protein) and VAMP (vesicle-associated membrane protein) types that have previously been localized to pantophysin vesicles in cultured cells. By immunofluorescence microscopy, pantophysin expression is seen predominantly in non-neuroendocrine cells with pronounced membrane traffic. Pantophysin staining codistributes with SCAMP and VAMP immunoreactivities in most instances but differs in some. Remarkably, pantophysin staining in liver is restricted to cells surrounding sinusoids and is not detectable in hepatocytes, similar to that of the SCAMP and VAMP isoforms as detected by our reagents in tissue sections.
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  • 24
    ISSN: 1432-0878
    Keywords: Key words Mammary gland ; Differentiation ; Neoplasia ; Homeodomain ; Cancer ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  In the mouse mammary gland, homeobox gene expression patterns suggest roles in development and neoplasia. In the human breast, we now identify a family of Iroquois-class (IRX) homeobox genes. One gene, IRX-2, is expressed in discrete epithelial cell lineages being found in ductal and lobular epithelium, but not in myoepithelium. Expression is absent from associated mesenchymal adipose stroma. During gland development, expression is concentrated in terminal end buds and terminal lobules and is reduced in a subset of epithelial cells during lactation. In contrast to observations for many homeobox genes in the mouse mammary gland in which homeobox gene expression is lost on neoplastic progression, IRX-2 expression is maintained in human mammary neoplasias. Data suggest IRX-2 functions in epithelial cell differentiation and demonstrate regulated expression during ductal and lobular proliferation as well as lactation.
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  • 25
    ISSN: 1432-0878
    Keywords: Key words Cytokine ; ELISA ; FACScan ; Lipids ; Macrophage ; Monocyte ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Diabetes (type I and type II) affects approximately 13 million people in the United States. Delayed and incomplete healing of wounds can be a major problem for diabetic patients. Macrophages are an important cell in the complex process of wound repair representing the major source of cytokines throughout the wound-healing process. Cytokines mediate many of the cellular responses critical to timely wound repair. It has been suggested that diabetes impairs wound healing through disruption of local cytokine production. Our previous in vivo studies in rats demonstrated that diabetes-induced and diet-induced hyperlipidemia cause changes in macrophage phenotype and function (Iacopino 1995; Doxey et al. 1998), suggesting that alterations in macrophage cytokine profiles represent the cellular/molecular mechanism responsible for delayed wound healing. The purpose of this study was to investigate how monocyte maturation/differentiation and cytokine production were altered by serum lipids in an in vitro system using human cells. Commercially prepared purified human monocytes were cultured and exposed to serum lipids. Phenotypic analysis of differentiated macrophages was then performed by flow cytometry and fluorescent microscopy using surface antigens specific for various macrophage subsets. Selected cytokines in conditioned medium were assayed using commercial human enzyme-linked immunosorbent assay (ELISA) kits. We demonstrate that serum lipids cause an increase in monocytic differentiation leading to an inflammatory macrophage phenotype rather than a reparative/proliferative phenotype. We also show that serum lipids cause a generalized decrease in macrophage cytokine production using interleukin-1 beta (IL-1β), tumor necrosis factor alpha (TNF-α), platelet-derived growth factor (PDGF), and transforming growth factor beta 1 (TGF-β1) as marker cytokines. Our present in vitro results using human cells confirm our previous in vivo studies in the rat and support the hypothesis that diabetes-induced hyperlipidemia alters the monocyte differentiation process resulting in changes of macrophage subsets and cytokine release at the wound site, ultimately impairing the wound-healing process.
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  • 26
    ISSN: 1432-0878
    Keywords: Key words Cultured epidermis ; Keratinocyte ; Epithelialization ; Wound healing ; Human ; Mouse (NMRI)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Sheets of cultured allogeneic human keratinocytes have been used for the treatment of burns and chronic leg ulcers but there has been no animal assay for the therapeutic action of these cultures. In order to analyze the effects of frozen cultures of human keratinocytes on wound healing, we have developed such an assay based on the rate of repair of full-thickness skin wounds in immunocompetent NMR1 mice. Reepithelialization of the control wounds, originating from the murine epithelium at the edge of the wound, occurred at a constant rate of advance of 150 µm/day. When frozen cultured human epidermal sheets were thawed at room temperature for 5–10 min and applied to the surface of the wound, the murine epithelium advanced at 267 µm/day. Most wounds treated with frozen cultures completely healed after 10 days, whereas most control wounds required 16 days. The accelerated reepithelialization did not depend on the presence of proliferative human keratinocytes in the frozen cultures. The cultures also promoted early formation of granulation tissue and laminin deposition over the surface of the wound bed. This simple assay should permit quantitative analysis of the effects on healing exerted not only by cultured cells, but also by proteins and small molecules.
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  • 27
    ISSN: 1432-0878
    Keywords: Key words Interleukin-18 ; Hyperosmotic conditions ; Epithelial cells ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Interleukin-18 is a novel multifunctional cytokine, which enhances natural killer cell activity and promotes the induction of cytokine production, including that of interferon-γ by T cells and antitumor effects. Interleukin-18 is produced by cells of several different tissues (e.g., macrophages, keratinocytes, osteoblasts, and intestinal epithelium); however, it is unclear what physiological conditions or stimuli induce interleukin-18 production. To determine physiological conditions for the production of interleukin-18, we have examined the effect of mannitol-induced hyperosmotic conditions on normal human umbilical vein endothelial cells (HUVEC) and eight established human epithelial-like cell lines (Intestine 407, Caco-2, A253, HeLa, SCC25, HT1197, ACHN, A549). Hyperosmotic conditions induced interleukin-18 immunoreactivity in all the human cell lines tested, as detected by immunocytochemistry. The enhanced interleukin-18 production was also observed when mannitol was replaced with NaCl as the inducer of hyperosmotic stress. Enzyme-linked immunosorbent assays revealed that interleukin-18 concentrations in cell extracts were significantly increased by hyperosmotic conditions. Reporter gene assays also revealed that hyperosmotic conditions stimulated transcriptional activity of the interleukin-18 promoter. These results show for the first time that hyperosmotic stress is a stimulator of interleukin-18 production in epithelial-like cells.
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  • 28
    ISSN: 1432-0878
    Keywords: Key words α1-Microglobulin ; Bikunin ; Inter-α-trypsin inhibitor ; Mast cells ; Trypstatin ; Urinary trypsin inhibitor ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  We recently reported that the rat mast cell proteinase inhibitor trypstatin is genetically identical with the second half of inter-α-trypsin inhibitor light chain (ITI-LC), also known as bikunin or urinary trypsin inhibitor (UTI). In this study, therefore, immunoreactivities of mast cells of various human tissues were examined with three antibodies, anti-human ITI-LC, anti-ITI, which recognizes mainly heavy chains or the sugar moiety of ITI, and anti-α 1-microglobulin (α1mG). ITI-LC immunoreactivity was strongly found in mast cells in the connective tissues of various organs except for those of the propria mucosae of small intestine. Neither anti-ITI antibody nor anti-α1mG antibody reacted with mast cells in various tissues. By reverse transcription-polymerase chain reaction (RT-PCR) analysis, α1mG/ITI-LC mRNA was not detected in the skin and tongue, and only weakly in small intestine, although ITI-LC immunoreactivity was strongly detected in these tissues. Furthermore, the mRNA was not expressed in cultured human mast cells. These results suggest that ITI-LC protein is stored in the granules of human connective tissue mast cells, though is not produced by them.
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  • 29
    ISSN: 1432-0878
    Keywords: Key words Osteoblasts ; Extracellular matrix ; Transforming growth factor-β1 ; Basic fibroblast growth factor ; Apert’s syndrome ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The phenotype of Apert osteoblasts differs from that of normal osteoblasts in the accumulation of macromolecules in the extracellular matrix. Apert osteoblasts increase type I collagen, fibronectin and glycosaminoglycans secretion compared with normal osteoblasts. Because the extracellular matrix macromolecule accumulation is greatly modulated by transforming growth factor-β1, we examined the ability of normal and Apert osteoblasts to secrete transforming growth factor-β1 by CCL-64 assay and to produce transforming growth factor-β1 by analysis of the mRNA expression of transforming growth factor-β1. Northern blot analysis revealed an increased amount of transforming growth factor-β1 mRNA expression in Apert osteoblasts compared with normal ones. Moreover, the level of the active transforming growth factor-β1 isoform was higher in Apert than in normal media. In pathologic cells, the increase in transforming growth factor-β1 gene expression was associated with a parallel increase in the factor secreted into the medium. The level of transforming growth factor-β1 was decreased by the addition of basic fibroblast growth factor. Transforming growth factor-β1 is controlled temporally and spatially during skeletal tissue development and produces complex stimulatory and inhibitory changes in osteoblast functions. We hypothesise that in vitro differences between normal and Apert osteoblasts may be correlated to different transforming growth factor-β1 cascade patterns, probably due to an altered balance between transforming growth factor-β1 and basic fibroblast growth factor.
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  • 30
    ISSN: 1432-0878
    Keywords: Key words Endothelin ; Endothelin receptors ; Endothelin-converting enzyme ; Ovary ; In situ hybridization ; Human ; Cynomolgus monkey
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The endothelin system is composed of three endothelin isoforms (ET-1, ET-2, and ET-3), the endothelin receptors ETA and ETB, and the endothelin-converting enzyme (ECE). Besides having a major vasoactive role, endothelins have roles in different cell types at a local level. We investigated the presence of the different components of the endothelin system in primate ovaries. Human ovaries and gonadotropin-stimulated monkey ovaries were studied using immunohistochemistry for endothelin, and in situ hybridization with probes for ET-1, ET-2, ET-3, ETA and ETB receptors, and ECE. ET-1 and ETA receptors were detected in endothelial cells and vascular smooth muscle cells, respectively, in stromal vessels adjacent to follicles and corpora lutea. ETB receptors and ET-1 were found in the endothelial cells of capillaries of corpora lutea. ECE was present in internal theca cells of secondary, de Graaf, atretic follicles, and in luteinized granulosa cells of the corpora lutea. The endothelin system components are present in or around the follicles of human and monkey ovaries. Although the components are not expressed in the same cell types, they are synthesized, mainly in follicles, by cells that are in close proximity. Thus, the endothelin system could act in a paracrine manner. ECE expression in steroid-producing cells changes its compartmentalization during follicle maturation.
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  • 31
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    Cell & tissue research 295 (1999), S. 297-305 
    ISSN: 1432-0878
    Keywords: Key words Trophoblast ; Macrophages ; Placenta ; Cell culture ; Paracrine regulation ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract In view of the accumulating evidence for paracrine mechanisms regulating trophoblast function, we tested the hypothesis that placental macrophages affect trophoblast activity in a paracrine fashion. Trophoblast was isolated from 17 term placentas (–IP). One aliquot of cells was further immunopurified (+IP) using an HLA class I antibody. This increased the proportion of trophoblast (+IP 〉97%; –IP ∼70%) as identified by rigorous immunocytochemistry. Most (∼70%) non-trophoblast cells in –IP were macrophages. The cells were cultured for 5 days with a daily medium change. In addition, +IP cells from seven placentas were cultured with lipopolysaccharide (LPS)-stimulated or -unstimulated macrophage-conditioned media. The concentrations of lactate, trophoblast-specific hormones, human chorionic gonadotropin-β (hCG-β) and human placental lactogen (hPL), of several prostanoids and of endothelin-1 and angiotensin II were determined in the culture media. The accumulated amounts of substances released into the culture media, corrected for the greater proportion of trophoblast in +IP cultures, were on average two- to threefold higher (hCG-β: 18-fold) in +IP than in –IP, with the exception of endothelin-1,2 (no change), angiotensin II (–70%) and 6-keto-prostaglandin-F1α (–40%). [3H]leucine incorporation into the trichloroacetic acid (TCA)-precipitable pool measured on day 5 was twofold higher in +IP than in –IP. Addition of conditioned media reverted these changes. The data demonstrate that placental macrophages in culture affect trophoblast biosynthetic activity in a paracrine fashion. We conclude that macrophages are important regulators of trophoblast activity.
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  • 32
    ISSN: 1432-0878
    Keywords: Key words Cerebrovascular development and injury ; Hemangioma ; Angiogenesis ; Immunocytochemistry ; Adhesion molecules ; Conventional transmission and high-voltage electron microscopy ; Mouse (C57BL ; SJL/J) ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Blood vessels from the vasculature of mouse brains during postnatal development and from human brain tumors (hemangiomas) removed at biopsy were examined immunocytochemically by transmission electron microscopy (TEM) or high-voltage transmission electron microscopy (HVEM) to determine the expression of intercellular adhesion molecule-1 (ICAM-1). In the mouse brains, ICAM-1 was shown to be initially expressed on the luminal and abluminal endothelial cell (EC) surfaces on day 3 after birth. ICAM-1 intensity increased on the luminal EC surfaces and labeled vesiculotubular profiles (VTS, defined in the present report) between days 5 and 7. After 2 weeks and at 6 months after birth, ICAM-1 labeling was weak or absent on the luminal EC surfaces. The hemangiomas presented a strong ICAM-1 reaction product on the luminal EC surfaces of small and large blood vessels associated with the VTS, with a weaker labeling of the abluminal or adventitial aspects of larger blood vessels. TEM of vesiculovacuolar structures (VVOs) within ECs from arteries and veins also demonstrated reaction product for ICAM-1 labeling. Three-dimensional stereo-pair images in the HVEM enhanced the visualization of gold particles that were attached to the inner-delimiting membrane surfaces of EC VTS, and VVOs, respectively. These observations raise the possibility that the neonatal leukocytes and tumor cells may utilize these endothelial structures as a route across the developing and injured blood-brain barrier (BBB).
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  • 33
    ISSN: 1432-0878
    Keywords: Key words Atherosclerosis ; Culture model ; Endothelial cell ; Endothelin-1 ; Oxidized LDL ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We investigated the structural and functional properties of human umbilical vein endothelial cells (HUVECs) cultured on a two-chamber culture model system using an amnion membrane. Compared to HUVECs cultured on a plastic dish, HUVECs cultured on the model system exhibited several features similar to those of in vivo vessels, including formation of the intercellular junctional devices and expression of tight junction-associated protein ZO-1 and adherence junction-associated protein α-catenin. Furthermore, we found that HUVECs had a property of polar secretion of endothelin-1 (ET-1). About 90% of the total amount of synthesized ET-1 was found in the lower well, designated as the basal side. When HUVECs were incubated with either native low-density lipoproteins (nLDLs) or oxidized LDLs (oxLDLs) at a concentration of 100 μg/ml, ET-1 secretion was significantly increased, dependent on the cell side (apical vs basal) on which the nLDLs or oxLDLs were loaded. When the LDLs were loaded on the apical side, the secretion of ET-1 from HUVECs on the apical side was increased by 48% (nLDL) and 61% (oxLDL), whereas it was accompanied by a concomitant decrease of ET-1 on the basal side (45% by nLDLs and 38% by oxLDLs). When loaded on the basal side, however, ET-1 was increased by 23% (nLDLs) and 53% (oxLDLs) on the basal side, with a 26% simultaneous decrease of ET-1 on the opposite side for both nLDLs and oxLDLs. On the contrary, high-density lipoproteins (HDLs) inhibited ET-1 secretion from HUVECs on the opposite side of the well on which HDLs were loaded; there was a 57% decrease on the basal side when HDLs were loaded on the apical side, and a 46% decrease on the apical side when loaded on the basal side. These results indicate that modulation of ET-1 secretion from ECs by lipoproteins is virtually dependent on the place (apical vs basal) where these proteins are present. The finding that nLDLs and oxLDLs enhance ET-1 secretion by ECs in a polarized pattern suggests that ET-1 may be involved in pathophysiological processes such as atherogenesis.
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  • 34
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    Cell & tissue research 295 (1999), S. 151-158 
    ISSN: 1432-0878
    Keywords: Key words Class II MHC-positive cells ; Human leukocyte antigen-DR ; Dental pulp ; Dendritic cells ; Macrophages ; Ultrastructure ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The distribution and ultrastructure of class II major histocompatibility complex (MHC)-positive cells were investigated in human dental pulp, employing immunohistochemistry using an anti-human leukocyte antigen (HLA)-DR-monoclonal antibody. HLA-DR-immunopositive cells, appearing spindle-like or dendritic in profile, were densely distributed throughout the dental pulp. Under the electron microscope, these cells exhibited various sizes of vesicles containing clear or opaque contents, multivesicular bodies and characteristic fine tubulovesicular structures in their cytoplasm. Some reactive cells possessed coated pits and vesicles including electron-dense materials, indicating an active endocytosis. At the periphery of the pulp tissue, the HLA-DR-immunopositive cells were predominantly situated in the subodontoblastic layer, with some located in the odontoblast layer and/or predentin and extending their cytoplasmic processes into the dentinal tubules. Cell processes of these cells occasionally made contact with several odontoblast processes in the same way as the nerve fibers in the predentin. These cells never contained the typical phagosomes frequently observed in the HLA-DR-immunoreactive macrophages in the subodontoblastic layer and the pulp core. The results suggest that the HLA-DR-immunopositive cells in the odontoblast layer and/or predentin have some regulatory function on the odontoblasts under physiological conditions, in addition to their involvement in the initial defense reaction after tooth injury.
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  • 35
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    International journal of salt lake research 8 (1999), S. 171-176 
    ISSN: 1573-8590
    Keywords: algae ; Lake Bai-Khol ; salinity ; Uvs-Nuur Basin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geography
    Notes: Abstract The algal flora of Lake Bai-Khol, located in the endorheic Uvs-Nuur Basin, Russia, has been studied from collections made in August 1997. 79 taxa belonging to diatoms, blue-green and green algae were present.
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  • 36
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    International journal of salt lake research 8 (1999), S. 127-138 
    ISSN: 1573-8590
    Keywords: Lake Elementaita ; Lake Nakuru ; origin ; salinity ; thermal springs ; water chemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geography
    Notes: Abstract The hydrochemistry of three areas of geothermal activity in the Lake Elementaita basin were surveyed. The results indicated warm temperatures between 30--40°C, high alkalinity (1,200 mg CaCO3/l) and specific conductance (4,300--8,100 µS/cm). pH was above 9 with a high concentration of carbonates, chlorides and sulphates. Apart from playing a significant role in the lake chemistry, lacustrine springs provide a significant source of water supply, particularly during the dry season. However, there is a serious lack of scientific information on the hydrogeology and hydrochemistry of the thermal springs.
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  • 37
    ISSN: 1573-8590
    Keywords: black bream ; estuaries ; fecundity ; fishery management ; length and age at maturity ; salinity ; spawning periods and locations ; water temperature
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geography
    Notes: Abstract The reproductive biology ofAcanthopagrus butcheri has been studied in the permanently open Swan River and intermittently open Moore River estuaries on the lower west coast of Australia (31–32°S) and in the permanently open Nornalup Walpole and normally closed Wellstead estuaries on the southern coast of Western Australia (34–35°S). Trends exhibited by gonadosomatic indices, gonadal maturity stages and the sizes and developmental stages of the oocytes demonstrate thatA. butcheri typically spawns in spring and early summer. However, spawning occurred in salinities ranging from as low as 3.5–8 gL−1 in the Moore River Estuary to as high as 41–45 g L−1 in the Wellstead Estuary. Furthermore, water temperatures during spawning were greater in the two northern estuaries (19.7–28.5°C) than in the two southern and cooler estuaries (17.5–23.4°C). Histological studies strongly indicate thatA. butcheri spawn more than once in a breeding season and demonstrate that the development of its oocytes exhibits group synchronysensu de Vlaming (1983). The ages and total lengths at which, on average, female and maleA. butcheri both first attain maturity in the Swan River Estuary were ca 2 years and ca 215 mm. However, the age at which individual fish in that system reach maturity was influenced by body size. This suggests that the attainment of first maturity at an older age but smaller length in the Moore River and Nornalup Walpole estuaries than is the case in the Swan River Estuary is a consequence of the slower growth rates ofA. butcheri in those estuaries. The combination of the young age (ca 2 years) but small length (ca 145 mm) at which maturity is first attained in the Wellstead Estuary could have resulted from selection pressures brought about by high mortality rates and/or heavy fishing pressure in this estuary. The mean fecundity ofA. butcheri, based on the combined number of yolk vesicle and yolk granule oocytes found in ovaries just prior to the onset of spawning, was 1580×103. The significance of the sizes at first maturity, minimum legal length for capture, mesh selectivity data and closure of certain regions of estuaries to fishing for the management of the recreational and commercial fishery forA. butcheri is discussed.
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  • 38
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    International journal of salt lake research 8 (1999), S. 293-306 
    ISSN: 1573-8590
    Keywords: Aral Sea ; salinity ; salinization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geography
    Notes: Abstract An overview of recent changes insalinity in the surface waters of the Aral sea basinis given. Total dissolved salts (salinity) in mostwaters are higher than admissible values fordrinking water, and ionic composition has changed withtime. Salinity of the `Big Sea' of the Aral was 48g L−1 in 1998, but has decreased to below 21g L−1 in the `Small Sea' due to the building ofa new dam between the two parts of the Aral Sea.
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  • 39
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    International journal of salt lake research 8 (1999), S. 127-138 
    ISSN: 1573-8590
    Keywords: Lake Elementaita ; Lake Nakuru ; origin ; salinity ; thermal springs ; water chemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geography
    Notes: Abstract The hydrochemistry of three areas of geothermal activity in the Lake Elementaita basin were surveyed. The results indicated warm temperatures between 30–40°C, high alkalinity (1,200 mg CaCO3/l) and specific conductance (4,300–8,100 μS/cm). pH was above 9 with a high concentration of carbonates, chlorides and sulphates. Apart from playing a significant role in the lake chemistry, lacustrine springs provide a significant source of water supply, particularly during the dry season. However, there is a serious lack of scientific information on the hydrogeology and hydrochemistry of the thermal springs.
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  • 40
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    International journal of salt lake research 8 (1999), S. 361-382 
    ISSN: 1573-8590
    Keywords: Aral Sea ; salinity ; salinization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geography
    Notes: Abstract Many streams in Western Australia are naturally saline. In others, especiallyin the south-western corner, land-clearance and other human activities inthe catchment have accelerated rates of salinisation of surface andgroundwater. Trends in surface water salinity are well-documented but theextent of penetration of saline stream water into the sediments has beenlittle studied. As many of these streams have porous sandy beds and theirflows may derive from groundwater, hydrologic exchange patterns betweensurface water and subsurface hyporheic water were hypothesised togovern the water chemistry of such rivers. We predicted high rates ofhydraulic conductivity, leading to a close relationship between surface andsubsurface (to a depth of 50 cm) salinity, and to a lesser extent, pH anddissolved oxygen. Where surface and hyporheic water differed in salinity,other chemical differences were hypothesised to be similarly marked,perhaps resulting from disjunct shallow subsurface aquifers. Triplicatewells were sampled from upwelling and downwelling zones of thirteenstreams ranging in salinity from ca. 0.2 to 18 g L−1. Despite theseemingly-porous sandy beds at many sites, subsurface water chemistryonly 20–40 cm below the bed sometimes differed markedly from surfacewater. For example, hyporheic water was only one-fifth the salinity ofsurface water at some saline sites (e.g., the Tone River) or 20 per cent moresaline in streams with fresh surface water (e.g., the Weld River). At somesites of intermediate salinity (e.g., the Warren River), subsurface water wasup to three times fresher than surface or downwelling water. Percentagesaturation of dissolved oxygen in the hyporheic water was consistently low(〈 40%) whereas pH was more acidic than surface water, presumably dueto microbial activity. Vertical hydraulic conductivity may be limited bylayers of fine sediments and clays, implying that the meso-scale (1–100 cm)hydrological dynamics within the hyporheic zones of these rivers are morecomplex than their sandy beds would indicate. Assumptions of ecosystemdynamics in saline streams must be tempered by an understanding ofhyporheic salinities as subsurface fresher water may support microbial andfaunal assemblages excluded from the surface benthos by high salinity. Insaline streams, as in fresh ones, the hyporheic zone is an importantcomponent of the stream ecosystem and equally prone to disruption byhuman activities.
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  • 41
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    International journal of salt lake research 8 (1999), S. 55-70 
    ISSN: 1573-8590
    Keywords: Dead Sea ; equation of state ; pycnometry ; salinity ; saturation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geography
    Notes: Abstract Some widespread misconceptions about salinity are pointed out, explained and commented on. Definitions, units, and methods of salinity determination recommended for brines of moderate salinities may become unsuitable for highly saline brines. Various salinity units are examined and commented on, and density at a precisely monitored temperature is chosen as the preferable “substitute parameter” for salinity determination of hypersaline brines. It is shown that ambient air density during pycnometric measurements must be known as accurately as required of the brine's density, and that its value must be added to the direct pycnometric measurement. Variations of atmospheric pressure at the time of pycnometry must be taken into account if greater than 15 mB. For a salinity accuracy of ±0.02 per mille, as required for some physical and chemical studies, the temperature of the sample during salinity measurement must be monitored with an accuracy of at least 0.04°C. A definite curve in the ρ-S plane, corresponding to the conversion of salinity to density at a fixed temperature, andvice-versa, does not exist if the brine is saturated in one of its salts. Non-linearity in the equation of state is shown to affect the surface level drop due to evaporation; the effect is negligible at low salinities, but a 10 per cent correction is already required at the salinity found in the northern Red Sea (41 per mille). Reliable reference points for the conversion of absolute salinity to density of highly saline brines are not known, at present, as accurately as desirable; pending the accurate determination of such reference points, a substantial dilution of hypersaline brines (for the purpose of adapting them to conductivity measurements) would multiply the error range by more than a hundred fold. On the other hand, a minute dilution of hypersaline brine samples to prevent salt crystal formation is possible and would not bring about any major increase of the error range in their density (salinity) determination.
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  • 42
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    International journal of salt lake research 8 (1999), S. 171-176 
    ISSN: 1573-8590
    Keywords: algae ; Lake Bai-Khol ; salinity ; Uvs-Nuur Basin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geography
    Notes: Abstract The algal flora of Lake Bai-Khol, located in the endorheic Uvs-Nuur Basin, Russia, has been studied from collections made in August 1997. 79 taxa belonging to diatoms, blue-green and green algae were present.
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  • 43
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    International journal of salt lake research 8 (1999), S. 55-70 
    ISSN: 1573-8590
    Keywords: Dead Sea ; equation of state ; pycnometry ; salinity ; saturation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geography
    Notes: Abstract Some widespread misconceptions about salinity are pointed out, explained and commented on. Definitions, units, and methods of salinity determination recommended for brines of moderate salinities may become unsuitable for highly saline brines. Various salinity units are examined and commented on, and density at a precisely monitored temperature is chosen as the preferable “substitute parameter” for salinity determination of hypersaline brines. It is shown that ambient air density during pycnometric measurements must be known as accurately as required of the brine's density, and that its value must be added to the direct pycnometric measurement. Variations of atmospheric pressure at the time of pycnometry must be taken into account if greater than 15 mB. For a salinity accuracy of ±0.02 per mille, as required for some physical and chemical studies, the temperature of the sample during salinity measurement must be monitored with an accuracy of at least 0.04°C. A definite curve in the ρ-S plane, corresponding to the conversion of salinity to density at a fixed temperature, and vice-versa, does not exist if the brine is saturated in one of its salts. Non-linearity in the equation of state is shown to affect the surface level drop due to evaporation; the effect is negligible at low salinities, but a 10 per cent correction is already required at the salinity found in the northern Red Sea (41 per mille). Reliable reference points for the conversion of absolute salinity to density of highly saline brines are not known, at present, as accurately as desirable; pending the accurate determination of such reference points, a substantial dilution of hypersaline brines (for the purpose of adapting them to conductivity measurements) would multiply the error range by more than a hundred fold. On the other hand, a minute dilution of hypersaline brine samples to prevent salt crystal formation is possible and would not bring about any major increase of the error range in their density (salinity) determination.
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  • 44
    ISSN: 1573-8590
    Keywords: black bream ; estuaries ; fecundity ; fishery management ; length and age at maturity ; salinity ; spawning periods and locations ; water temperature
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geography
    Notes: Abstract The reproductive biology of Acanthopagrus butcheri has been studied in the permanently open Swan River and intermittently open Moore River estuaries on the lower west coast of Australia (31--32 °S) and in the permanently open Nornalup Walpole and normally closed Wellstead estuaries on the southern coast of Western Australia (34--35 °S). Trends exhibited by gonadosomatic indices, gonadal maturity stages and the sizes and developmental stages of the oocytes demonstrate that A. butcher typically spawns in spring and early summer. However, spawning occurred in salinities ranging from as low as 3.5-8 g L−1 in the Moore River Estuary to as high as 41-45 g L−1 in the Wellstead Estuary. Furthermore, water temperatures during spawning were greater in the two northern estuaries (19.7--28.5 °C) than in the two southern and cooler estuaries (17.5--23.4 °C). Histological studies strongly indicate that A. butcheri spawn more than once in a breeding season and demonstrate that the development of its oocytes exhibits group synchrony sensu de Vlaming (1983). The ages and total lengths at which, on average, female and male A. butcheri both first attain maturity in the Swan River Estuary were ca 2 years and ca 215 mm. However, the age at which individual fish in that system reach maturity was influenced by body size. This suggests that the attainment of first maturity at an older age but smaller length in the Moore River and Nornalup Walpole estuaries than is the case in the Swan River Estuary is a consequence of the slower growth rates of A. butcheri in those estuaries. The combination of the young age (ca 2 years) but small length (ca 145 mm) at which maturity is first attained in the Wellstead Estuary could have resulted from selection pressures brought about by high mortality rates and/or heavy fishing pressure in this estuary. The mean fecundity of A. butcheri, based on the combined number of yolk vesicle and yolk granule oocytes found in ovaries just prior to the onset of spawning, was 1580 × 103. The significance of the sizes at first maturity, minimum legal length for capture, mesh selectivity data and closure of certain regions of estuaries to fishing for the management of the recreational and commercial fishery for A. butcheri is discussed.
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  • 45
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    International journal of salt lake research 8 (1999), S. 293-306 
    ISSN: 1573-8590
    Keywords: Aral Sea ; salinity ; salinization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geography
    Notes: Abstract An overview of recent changes in salinity in the surface waters of the Aral sea basin is given. Total dissolved salts (salinity) in most waters are higher than admissible values for drinking water, and ionic composition has changed with time. Salinity of the ‘Big Sea’ of the Aral was 48 g L−1 in 1998, but has decreased to below 21 g L−1 in the ‘Small Sea’ due to the building of a new dam between the two parts of the Aral Sea.
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  • 46
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    Mycopathologia 146 (1999), S. 33-41 
    ISSN: 1573-0832
    Keywords: entomopathogenic hyphomycete ; vector control ; Chagas disease ; Beauveria bassiana ; Rhodnius prolixus ; humidity ; temperature ; incubation time ; mortality
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The impact of relative humidity (RH) on the infective potential of the isolate Bb INRA 297 of Beauveriabassiana (Bals.) Vuillemin (Deuteromycotina: Hyphomycetes) against first in star nymphs of Rhodnius prolixus Stål. (Hemiptera: Reduviidae) was determined. Fungus-treated insects were exposed to RHs ranged from 75 to 100% at 25 °C. Results clearly showed a threshold of humidity at ca. 96% for high and rapid mortality. After initial exposure to increasing periods of 97% (4, 8, 16, 24, 36 and 48 h) and subsequent transfer to constant lower RHs (43, 53,75 and 86%) at a constant 25 °C, an incubation of at least 48 h at 97% RH was necessary to kill all insects. On changing RHs of 97/75% and different regimes of temperature (15/28 °C,20/25 °C, 25/28 °C,and 25/35 °C), at least 72 h of initial exposure at 97% RH for the 15/28 °C regime, 48 h for the 20°/25°C and25/28 °C regimes and 36 h for25/35 °C were needed to kill all insects over a 6-day incubation time. Delayed exposure to favorable moisture condition (97% RH), significantly affected infection for up to a 3-day delay within the various temperature-humidity regimes tested.
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  • 47
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    Bulletin of experimental biology and medicine 127 (1999), S. 483-484 
    ISSN: 1573-8221
    Keywords: chorionic gonadotropin ; irradiation ; blood ; liver
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Chorionic gonadotropin exerts a protective effect on irradiated animals: it increases survival rate and number of peripheral blood leukocytes, improves the structure of liver parenchyma, and increases body weight.
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  • 48
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    Bulletin of experimental biology and medicine 127 (1999), S. 587-589 
    ISSN: 1573-8221
    Keywords: liver ; CCl 4 ; cirrhosis ; bone marrow ; hemopoiesis ; macrophages ; tumor necrosis factor-α
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract During CCl4-induced liver cirrhosis, cells of the hemopoiesis-inducing microenvironment in the bone marrow of BALB/c mice produced activity inhibiting the growth of erythropoiesis and granulomonocytopoiesis precursors. Stimulation with yeast polysaccharide zymosan increased the inhibitory activity (especially in relation to granulomonocytic precursors). The highest growth-inhibitory activity was produced by the bone marrow adherent fraction (residual bone marrow macrophages). Tumor necrosis factor-α is probably responsible for the inhibition of the growth of myeloid precursors in mice with CCl4-induced liver cirrhosis.
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  • 49
    ISSN: 1573-8221
    Keywords: liver ; cholestasis ; biotransformation of xenobiotics ; antioxidant system
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract An 8-day biliary obstruction in rats caused by ligation of the common bile duct is accompanied by an increase in bilirubin blood content and alanine aminotransferase activity. The hepatic xenobiotic-metabolizing activity (intensities of microsomal oxidation and glucuronidation) decreases in parallel with the inhibition of antioxidant enzymes (catalase, superoxide dismutase, and glutathione peroxidase). A comparative study of Heptral, Ursofalk, Essentiale, and Cordiamin (nikethamide) showed that Cordiamin displays the most significant enzymestabilizing activity.
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  • 50
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    Bulletin of experimental biology and medicine 128 (1999), S. 903-905 
    ISSN: 1573-8221
    Keywords: liver ; cirrhosis ; bone marrow ; hemopoiesis ; macrophages
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract During late stages of CCl4-induced liver cirrhosis, production of factors stimulating the growth of erythroid and granulomonocytopoietic precursors by bone marrow adherent and nonadherent fractions of BALB/c mice decreased. Stimulation with the yeast polysaccharide zymosan decreased production of these activities by various bone marrow cells, especially by the adherent fraction.
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  • 51
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    Immunogenetics 48 (1998), S. 40-46 
    ISSN: 1432-1211
    Keywords: Key words Immunoglobulin ; Genes ; Kappa light chain ; Human ; Antibody repertoire
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  We describe a hitherto unknown functional IGKV gene, VkLa, belonging to the IGKV1 subgroup with exon 2 having only 94% similarity to the closest known IGKV gene, 1–13/1D-13 (L4/L18a). Genomic DNA sequences spanning from 5’ of the decanucleotide box to 3’ of the heptamer (649 bp) were cloned and sequenced from four individuals. The new gene encodes the conserved amino acids in the exons and contains no apparent defects in known regulatory intron sequences such as pd-box, dc-box, TATA-box, CCCT-elements, splice-sequences, initiation codon, and heptamer sequence. VkLa is therefore potentially functional and, correspondingly, we found transcripts of properly rearranged VkLa with somatical hypermutations. VkLa was found in 12 of 57 (21%) healthy Caucasians by a nested polymerase chain reaction and subsequent sequencing of exon 2. This finding shows that there is more inter-individual variation in the available IGKV gene repertoire than was hitherto assumed. Finally, we describe a minor correction in the IGKV1D-43 (L23) gene sequence.
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  • 52
    ISSN: 1432-1211
    Keywords: Key words Antigen presenting cells ; Dendritic cells ; Cell surface molecule ; Antigen receptor ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Dendritic cells (DC) are specialist antigen presenting cells which capture antigens in the periphery, migrate centrally, and present the processed antigens in the context of major histocompatibility complex and appropriate co-stimulatory molecules to T lymphocytes for the initiation of an immune response. DEC-205 has been identified as a putative antigen-uptake receptor, which is expressed abundantly on mouse DC. The recently cloned mouse DEC-205 cDNA predicts a molecular structure which has a marked similarity to the macrophage mannose receptor. Using reverse transcriptase-polymerase chain reaction (RT-PCR) and cDNA library screening, we obtained the full coding region of human DEC-205 cDNA from the Hodgkin’s disease-derived L428 cell line. The predicted protein structure is a type I transmembrane protein of 1722 amino acids consisting of a signal peptide, cysteine-rich domain, fibronectin type II domain, ten carbohydrate recognition-like domains, transmembrane domain, and a cytoplasmic tail. Human DEC-205 is 77% identical to the mouse protein with completely conserved cysteines. The DEC-205 gene (LY75) was mapped to chromosome band 2q24 by somatic cell hybrid panel analysis and fluorescent in situ hybridization. Northern blot analysis detected 7.8 and 9.5 kilobase DEC-205 transcripts in myeloid, B lymphoid, and Hodgkin’s disease-derived cell lines. RT-PCR analysis indicated that immature blood DC contain a barely detectable amount of DEC-205 transcripts but these were markedly increased upon differentiation/activation.
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  • 53
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    International journal of biometeorology 42 (1998), S. 84-88 
    ISSN: 1432-1254
    Keywords: Key words Mortality ; Human ; Seasonality ; Secular trend ; Temperature
    Source: Springer Online Journal Archives 1860-2000
    Topics: Geography , Physics
    Notes: Abstract  Based on records from the Federal Bureau for Statistics of Germany, the seasonality of mortality was investigated for the period 1946–1995. Lowest mortality rates were found during summer (August or September) while highest values were found in winter (January through March). Non-linear regression of all monthly mortality data with the average monthly temperatures in Germany revealed a significant negative relationship (r=–0.739; n=600; P〈0.0001). The fact that the differences between the long-range monthly temperatures and the individual monthly temperatures also showed a distinct relationship to the mortality rates speaks against a mere coincidence of both parameters. The amplitude of this seasonal rhythm declined steadily within the observation period. It is concluded that low temperatures cause an increase in mortality rates and that this effect has become less important during recent decades due to the increased use of central heating and because of improvements in the public health system.
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  • 54
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    Immunogenetics 48 (1998), S. 312-323 
    ISSN: 1432-1211
    Keywords: Key words Recombination ; Gene conversion ; Allele ; MHC ; HLA ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Some alleles of the major histocompatibility complex (MHC) genes have a reticulate pattern of evolution, probably resulting from the exchange of segments by gene conversion or recombination. Here we compare the extent and patterns of reticulate evolution among the classical class I and class II loci of the human MHC using the recently developed compatibility and partition matrix methods. A complex pattern is revealed with substantial differences among loci in the extent and pattern of reticulation. Extremely high levels of reticulation are observed at HLA-B and HLA-DPB1, high levels at HLA-A and HLA-DRB1, moderate levels at HLA-C and HLA-DQB1, and low levels at HLA-DQA1. The reticulate events are concentrated in the exons encoding the highly variable, peptide-binding domains, suggesting that the sequence combinations produced by these events are maintained by natural selection.
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  • 55
    ISSN: 1432-1041
    Keywords: Key words CYP2D6 ; Genetic polymorphism ; Metoprolol ; Pharmacokinetics ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Abstract Aim: The aim of the present paper was to compare the pharmacokinetics of metoprolol in homozygous Caucasian volunteers for the wild-type CYP2D6 allele (CYP2D6*1/CYP2D6*1) and heterozygous (CYP2D6*1/CYP2D6*4) Caucasians. Methods: Thirty-six unrelated healthy male Caucasians were screened for two of the most frequently occurring mutant alleles (CYP2D6*3 and CYP2D6*4) using polymerase chain reaction (PCR). Twenty-four volunteers with a genotype suggesting a rapid hydroxylator phenotype were enrolled in a bioequivalence trial and each received in a randomized, cross-over fashion one of the three formulations compared. Each formulation contained 200 mg metoprolol tartrate/(tablet). In each of the three periods of the trial, one of the formulations was administered under fasting conditions in the morning on 4 consecutive days. Blood for quantification of metoprolol was drawn immediately before the last dose and in selected time intervals thereafter. A sensitive and specific high-performance liquid chromatography (HPLC) method with fluorescence detection was applied for the quantification of metoprolol. Pharmacokinetic parameters were determined for each subject and statistically compared in two groups of 16 homozygous (CYP2D6*1/CYP2D6*1) and six heterozygous (CYP2D6*1/CYP2D6*4) volunteers. Results: Significant differences between homozygous and heterozygous individuals were observed for all pharmacokinetic parameters. The AUC in the course of one those interval of 24 h (AUCτ), minium steady-state concentration (Cmin ss) and average steady-state concentration (Cav ss) values for heterozygous individuals were more than twice those of individuals. Significantly higher values for Css max , t1/2, half-value duration (HVD) and mean residence time (MRT) were also observed in heterozygous volunteers. The higher concentrations of metoprolol in heterozygous individuals also had pharmacodynamic consequences, namely, greater heart rate and blood pressure reduction.
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  • 56
    ISSN: 1573-4919
    Keywords: vitamin-A ; cellular retinol-binding protein ; liver ; hepatic stellate cells ; lipocytes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Liver is a major site of retinoid metabolism and storage, and more than 80% of the liver retinoids are stored in hepatic stellate cells. These cells represent less than 1% of the total liver protein, reaching a very high relative intracellular retinoid concentration. The plasma level of retinol is maintained close to 2 μM, and hepatic stellate cells have to be able both to uptake or to release retinol depending upon the extracellular retinol status. In view of their paucity in the liver tissue, stellate cells have been studied in primary cultures, in which they loose rapidly the stored lipids and retinol, and convert spontaneously into the activated myofibroblast phenotype, turning a long-term study of their retinol metabolism impossible. We have analyzed the retinol metabolism in the established GRX cell line, representative of stellate cells. We showed that this cell line behaves very similarly, with respect the retinol uptake and release, to primary cultures of hepatic stellate cells. Moreover, we showed that the cellular retinol binding protein (CRBP-I) expression in these cells, relevant for both uptake and esterification of retinol, responds to the extracellular retinol status, and is correlated to the retinol binding capacity of the cytosol. Its expression is not associated with the overall induction of the lipocyte phenotype by other agents. We conclude that the GRX cell line represents an in vitro model of hepatic stellate cells, and responds very efficiently to wide variations of the extracellular retinol status by autonomous controls of its uptake, storage or release.
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  • 57
    ISSN: 1573-4919
    Keywords: chicken ; oviduct ; liver ; tissue-specific repression ; in vivo gene transfer ; gene gun
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract In order to search tissue-specific elements in the 5′-upstream promoter region, gene gun was used to transfect in vivo plasmid DNAs with varying lengths of truncated ovalbumin promoter fused to the CAT reporter gene to the oviduct and liver of laying hens. The results indicated that in the oviduct, consistently high reporter gene expression was observed irrespective of the length of the truncated ovalbumin gene promoters, whereas in the liver the ovalbumin promoter extending from -3200 to +8 bp suppressed substantially the reporter gene expression compared with consistently high gene expression obtained by the ovalbumin promoters from -2800 to +8 bp or shorter length. It was concluded, therefore, that a tissue-specific silencer-like element might reside most likely in the ovalbumin gene promoter region between -3200 and -2800 bp which represses the ovalbumin gene transcription in the liver, but not in the oviduct of laying hens.
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  • 58
    ISSN: 1573-4919
    Keywords: trans polyunsaturated fatty acid ; linoleic acid ; desaturation elongation ; microsomes ; liver
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Several nutritional studies have shown the in vivo conversion of the 9c,12t-18:2 and 9t,12c-18:2 into long chain polyunsaturated fatty acids (PUFA) containing 20 carbons (geometrical isomers of eicosadienoic and eicosatetraenoic acids). In the present work, some in vitro studies were carried out in order to have precise information on the conversion of these two isomers. In a first set of experiments, studies were focused on the in vitro Δ6 desaturation, the first regulatory step of the biosynthesis of n-6 long chain PUFA, from 9c,12c-18:2. Rat liver microsomes were prepared and incubated under desaturation conditions with [1-14C]-9c,12c-18:2 in presence of unlabelled 9c,12t-, 9t,12c- or 9t,12t-18:2. The data show that each trans isomer induced a decrease of the Δ6 desaturation of the [1-14C]-9c,12c-18:2, but the 9c,12t-18:2 was the most potent inhibitor (up to 63%). Rat liver microsomes were also incubated with [1-14C]-9c,12c-18:2, [1-14C]-9c,12t-18:2 or [1-14C]-9t,12c-18:2 under desaturation conditions. The results indicated that 18:2 Δ9c,12t is a much better substrate for desaturase than 9t,12c-18:2. Moreover, the conversion levels of [1-14C]-9c,12t-18:2 was similar to what was observed for its all cis homologue, at low substrate concentration only. In a second set of experiments, in vitro elongation studies of each mono-trans 18:2 isomers and 9c,12c-18:2 were carried out. For that purpose, rat liver microsomes were incubated with [1-14C]-9c,12c-18:2, [1-14C]-9c,12t-18:2 or [1-14C]-9t,12c-18:2 under elongation conditions. The data show that [1-14C]-9t,12c-18:2 is better elongated than 9c,12c-18:2 while the amount of product formed from [1-14C]-9c,12t-18:2 was lower than was produced from the 9c,12c-18:2. Thus, the desaturation enzymes presented a higher affinity for the 9c,12t-18:2 whereas the elongation enzyme presented a higher affinity for the 9t,12c-18:2.
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  • 59
    ISSN: 1573-4919
    Keywords: carnitine palmitoyl transferase I ; mitochondrial HMG-CoA synthase ; dexamethasone ; suckling rats ; ketogenesis ; intestine ; liver
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The influence of the injection of dexamethasone on ketogenesis in 12 day old suckling rats was studied in intestine and liver by determining mRNA levels and enzyme activity of the two genes responsible for regulation of ketogenesis: carnitine palmitoyl transferase I (CPT 1) and mitochondrial HMG-CoA synthase. Dexamethasone produced a 2 fold increase in mRNA and activity of CPT I in intestine, but led to a decrease in mitt HMG-CoA synthase. In liver the mRNA levels and activity of both CPT I and mitt HMG-CoA synthase decreased. Comparison of these values with the ketogenic rate of both tissues following dexamethasone treatment suggests that mitt HMG-CoA synthase could be the main gene responsible for the regulation of ketogenesis in suckling rats. The changes produced in serum ketone bodies by dexamethasone, with a profile that is more similar to the ketogenic rate in the liver than that in the intestine, indicate that liver contributes more to ketone body synthesis in suckling rats. Two day treatment with dexamethasone produced no change in mRNA or activity levels for CPT I in liver or intestine. While mRNA levels for mitt HMG-CoA synthase changed little, the enzyme activity is decreased in both tissues.
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  • 60
    ISSN: 1432-0878
    Keywords: Key words Salivary gland ; Secretion ; Granule docking ; Exocytosis-endocytosis coupling ; OsO4 maceration ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The luminal membrane of salivary acinar cells creates a specialized cell surface area that accepts exocytosis and undergoes dynamic changes during secretion. These changes were visualized three-dimensionally from both the inside and outside of the cell in human parotid and submandibular glands, by application of in vitro secretory stimulation and then of OsO4 maceration to remove cytoplasmic organelles by varying degrees. In control glands treated without secretagogues, the luminal surface of serous acinar cells bore well-developed microvilli with only an occasional incidence of exocytotic profiles. Following treatment with the β-adrenergic agonist, isoproterenol, considerable shortening and loss of microvilli occurred along the luminal membrane where, on its cytoplasmic side, many protuberances of sizes similar to or smaller than those of single secretory granules (∼1 μm in diameter) appeared. The cytoplasmic surface of these protuberances exhibited small vesicles (∼100–150 nm in diameter) that, by transmission electron microscopy, were shown to be coated pits or vesicles present on or around the exocytosed granule membranes. Treatment of tissues with the muscarinic agonist carbachol also caused a decrease of microvilli and the appearance of protrusions at the luminal membrane. However, unlike isoproterenol treatment, many of these protrusions were devoid of small pits or vesicles and were much larger than a single secretory granule. These results indicate that (1) secretory stimulation causes the dynamic transformation of microvilli at the luminal membrane, where granule docking and membrane fusion take place, and (2) after fusion, the exocytosed membranes are processed differently, by coated pit/vesicle mediated or non-mediated mechanisms, according to the autonomic receptor control.
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  • 61
    ISSN: 1432-0878
    Keywords: Key words: Neurofilaments ; Phosphorylation ; Differentiation ; Immunocytochemistry ; Brain storage ; Fixation ; Microwave ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Neurofilaments, which are exclusively found in nerve cells, are one of the earliest recognizable features of the maturing nervous system. The differential distribution of neurofilament proteins in varying degrees of phosphorylation within a neuron provides the possibility of selectively demonstrating either somata and dendrites or axons. Non-phosphorylated neurofilaments typical of somata and dendrites can be visualized with the aid of monoclonal antibody SMI 311, whereas antibody SMI 312 is directed against highly phosphorylated axonal epitopes of neurofilaments. The maturation of neuronal types, the development of area-specific axonal networks, and the gradients of maturation can thus be demonstrated. Optimal immunostaining with SMI 311 and SMI 312 is achieved when specimens are fixed in a mixture of paraformaldehyde and picric acid for up to 3 days and sections are incubated free-floating. Neurons, with their dendritic domains immunostained by SMI 311 in a Golgi-like manner, can be completely visualized in relatively thick sections. The limitations of Golgi-preparations, such as glia-labeling, artifacts, and the staining of only a small non-representative percentage of existing neurons, are not apparent in SMI preparations, which additionally provide the possibility of selectively staining axonal networks. The results achieved in normal fetal brain provide the basis for studies of developmental disturbances.
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  • 62
    ISSN: 1432-0878
    Keywords: Key words mRNA ; Cancerous epithelium ; Autocrine growth regulation ; In situ hybridization ; Immunohistochemistry ; Western blotting ; Benign prostate hyperplasia ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Insulin-like growth factors (IGFs) are potent mitogens for a variety of cancer cells in vitro. A paracrine/autocrine role of IGF-II in the growth of breast and prostate cancer cells has been suggested. Information on cell-type-specific IGF-II expression in vivo in the breast and prostate is, however, limited. Thus, cell types expressing IGF-II mRNA and protein in tumors were identified by in situ hybridization and immunohistochemistry. Of 36 prostate, 17 breast, and 10 bladder cancers, and 9 paraganglioma tissues examined, IGF-II was expressed in more than 50% of prostate, breast, and bladder tumors, and in 100% of paraganglioma tumors. Expression levels of IGF-II were highest in the paraganglioma and bladder followed by prostate and breast tumors. In all the tumors expressing IGF-II, both mRNA and protein were localized to malignant cells, expression in the stroma being minimal. Since previous studies had indicated that an incompletely processed form of 15-kDa IGF-II exhibited higher mitogenic potency than the completely processed 7.5-kDa IGF-II form, the quantity and size of IGF-II proteins expressed in these tumors were analyzed by Western immunoblotting. Greater expression of 15-kDa IGF-II relative to the 7.5-kDa IGF-II form was clearly demonstrated in all six prostate cancers and in half of the two breast and four bladder cancers examined. The results are consistent with the hypothesis that the 15-kDa form of IGF-II expressed in cancerous cells contributes to autocrine cancer cell growth in vivo.
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  • 63
    ISSN: 1432-0878
    Keywords: Key words Alkaline phosphatase ; Caco-2 intestinal epithelial cells ; Differentiation ; Dipeptidyl dipeptidase ; Proliferation ; Tyrosine kinase ; Tyrosine phosphoproteins ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Intestinal epithelial cell differentiation is closely regulated during normal cell renewal, maturation, and malignant transformation. Since tyrosine phosphorylation influences differentiation in other cell types and has been reported to vary between crypt cells to differentiated villus tip cells, we investigated the influence of tyrosine phosphorylation in colonocyte differentiation, by using human colonic Caco-2 cells as a model and expression of the brush border enzymes alkaline phosphatase (AKP) and dipeptidyl peptidase (DPDD) as differentiation markers. We studied three tyrosine kinase inhibitors with different modes of action and specificities, viz., genistein, erbstatin analog (EA), and tyrphostin, and the tyrosine phosphatase inhibitor sodium orthovanadate. AKP- and DPDD-specific activities were assayed in protein-matched cell lysates by synthetic substrate digestion. We also correlated the effects of these agents on brush border enzyme activity with tyrosine phosphorylation of phosphoproteins by Western blotting. Genistein (5–75 mg/ml) dose-dependently stimulated AKP and DPDD with a maximal stimulation at 75 mg/ml by 158.6± 17.5% and 228.6±37.1% of control values, respectively (n=12, P〈0.001). The inactive analog genistin had no effect. Tyrphostin (25 mM) similarly stimulated AKP and DPDD by 138.6±6.6% and 131.8±1.5% of control values (n=12, P〈0.001). Unexpectedly, EA (0.1–10 mM) had the opposite effect, inhibiting AKP- and DPDD-specific activity significantly at 10 mM with a maximal 14.8±6.4% and 26.5±2.5% of control values (n=12, each P〈0.001). Sodium orthovanadate had a discordant effect on these two differentiation markers. Orthovanadate dose-dependently increased AKP to a maximal 188.5±16.1% of basal activity at 1.5 mM but decreased DPDD activity at 1.5 mM to 47.2±3.8% (n=9, P〈0.001 each). The effects of each agent were preserved when proliferation was blocked with mitomycin C, suggesting that the modulation of phenotype by these agents was independent of any effects of proliferation. The tyrosine phosphorylation of several phosphoprotein bands was affected differently by these agents. In particular, the tyrosine phosphorylation of one 70-kDa to 71-kDa band was increased by genistein and tyrophostin but deceased by EA. The different effects of these modulators of tyrosine kinase activity raise the possibility that at least two independent enzymes or pathways regulating tyrosine phosphorylation modulate intestinal epithelial differentiation. Furthermore, tyrosine phosphorylation of the 70-kDa to 71-kDa phosphoprotein may be important in the intracellular signaling by which intestinal epithelial cell differentiation is controlled.
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  • 64
    ISSN: 1432-0878
    Keywords: Key words Vinculin ; Talin ; Integrin ; Dystrophin ; Spectrin ; T-tubule ; Costamere ; Basal membrane ; Cardiac muscle cell ; Dilated cardiomyopathy ; Human ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The transverse tubule system of the cardiomyocyte remains undeformed despite the extreme forces it undergoes during the contraction-relaxation cycle, but the morphological basis for its stability remains unclear. Therefore, we have investigated the architecture and subcellular protein scaffold of the cardiac T-tubules and compared it with that of the costameres and of the free sarcolemma. Tissue samples from normal rat and monkey hearts, and left ventricular tissue from normal and cardiomyopathic human hearts obtained at transplantation surgery were investigated using immunocytochemistry and confocal microscopy and by electron microscopy. In addition, we used a re-differentiation model of isolated, cultured adult rat cardiomyocytes. The cell membrane of the cardiac T-tubules was found to contain the cell-matrix focal adhesion molecules (FAMs) vinculin, talin, the α5β1 integrin and the membrane-associated proteins (MAPs) dystrophin and spectrin. FAMs and MAPs were localized in the T-tubular membrane in a similar pattern: in longitudinally oriented myocytes as transverse punctate lines at the Z-level; in transversally cut myocytes a radial tubular network was found to extend throughout the interior of the cell. Immunolabeling for basement membrane components including collagen IV, fibronectin and laminin showed a colocalization with FAMs and MAPs parallel to the transverse T-tubules. The costameres of the sarcolemma showed a protein composition resembling that of the T-tubules but the intervening segments of free sarcolemma showed absence of FAMs and presence of MAPs. For the first time, we demonstrate the existence and protein composition of the T-tubular scaffold in the human heart. Furthermore, we show that cardiomyocytes from human failing hearts have less abundant but more dilated T-tubules than do experimental animals. These results indicate that the cardiac T-tubular system contains a subcellular scaffold closely resembling that of the costameres. It consists of FAMs, MAPs and basal lamina proteins that confer structural integrity to the cardiac T-tubular membrane during contraction/relaxation cycles.
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  • 65
    ISSN: 1432-0878
    Keywords: Key words Prion protein (PrPc) ; Electron microscopy ; Secretory granules ; Membrane ; Extracerebral tissues ; Hamster ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  In transmissible spongiform encephalopathies (TSE), such as scrapie in animals and Creutzfeldt-Jakob disease in humans, the central event is the conversion of a host-encoded amyloidogenic protein (PrPc) into an abnormal isoform (PrPsc) that accumulates as amyloid in TSE brain. PrPc is a membrane sialoglycoprotein synthesized in the central nervous system and elsewhere. We have examined the ultrastructural localization of PrPc in numerous hamster and some human extracerebral tissues, by means of a post-embedding electron-microscopic method combined with immunogold labeling. In stomach, intestine, lung, and kidney from hamsters, and in stomach, kidney, and spleen from humans, immunogold labeling specific for PrPc is observed on various cellular substructures related to secretory pathways: Golgi apparatus, secretory globules, and plasma membrane. In mucous epithelial cells of stomach and intestine, PrPc appears to be concentrated in secretory globules, suggesting a role for PrPc in the secretory function of the digestive tract. The secretory aspect of PrPc may be a key to understanding the physiopathological mechanisms underlying TSE.
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  • 66
    ISSN: 1432-0878
    Keywords: Key words Neurons ; Brain ; Chemotaxis ; Endothelial cells ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The formylpeptide receptor (FPR), previously found only on polymorphonuclear leukocytes and monocytes/macrophages, responds to both synthetic N-formyl oligopeptides and those produced by bacteria. The cDNA for human FPR has been cloned and a rabbit polyclonal antiserum directed against a synthetic 11-amino-acid peptide corresponding to the deduced carboxy-terminus has been produced. We have now extensively characterized and used the antibody to detect FPR on normal human tissues and cell types. The receptor antigen is present on some epithelial cells, especially those with a secretory function, and on some endocrine cells, e.g., follicular cells of the thyroid and cortical cells of the adrenal. Liver hepatocytes and Kupffer cells are positive. Smooth muscle and endothelial cells are also generally positive. In the brain and spinal cord, the neurons of the motor, sensory, and cerebellar systems, and those of the parasympathetic and sympathetic systems stain positively. These data suggest that the putative endogenous agonist for FPR or an antigenically similar receptor reacts with cellular targets in the neuromuscular, vascular, endocrine, and immune systems.
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  • 67
    ISSN: 1432-0878
    Keywords: Key words Organotypic cocultures ; Histodifferentiation ; Proliferation ; Basement membrane components ; Integrins ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Cell-matrix interactions and the ordered deposition of basement membrane (BM) components are of major importance for the maintenance of tissue homeostasis in complex epithelia. This aspect was studied in vitro in a coculture system designed as an oral mucosa model. As crucial epithelial features the kinetics of proliferation, expression of site-specific keratins as well as integrin patterns in correlation to synthesis of BM components were assessed by immunohistochemistry and in situ hybridization. Comparison with non-cornified gingiva as tissue of origin revealed different stages of epithelial development, eventually leading to complete reconstruction within a time frame of 1–3 weeks. First, the initial activated stage up to 1 week was characterized by (a) high keratinocyte proliferation, (b) extended expression of the basal cell-specific keratin K5 and (c) a patchy pattern of the differentiation-specific keratins K4 and K13. Second, after 2 weeks the improvement of histoarchitecture correlated to (a) predominant K5 expression in the basal and (b) extension of K4 and K13 within the suprabasal cell compartment, (c) high expression of integrins α3β1 and α6β4 including their ligand laminin-5 and (d) accumulating deposition of basement membrane components. Third, virtually complete tissue normalization at 3 weeks was indicated by (a) restriction of K5 to the basal cell area, (b) regular suprabasal localization of K4 and K13, (c) polarization of integrins to basal and parabasal cells and (d) linear codistribution of collagen IV, “classical” laminin (-1 or -10) and laminin-5 underneath the basal cells. Thus, these organotypic cocultures represent relevant equivalents for non-keratinized oral mucosa with typical gingival differentiation features and in addition suitable models for preclinical trials such as prospective dental material testing.
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  • 68
    ISSN: 1432-0878
    Keywords: Key words Airways ; Mucins ; Secretion ; Mini-organ culture ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  To study the secretory products and the proliferation of cells of the human respiratory surface epithelium, we established a miniorgan-culture system of bronchial tissue. Biopsies of large airways were grown on agar-coated dishes immersed in a serum-enriched medium. As determined by light and transmission electron microscopy, between 1 and 3 weeks, the organ cultures were covered by a differentiated epithelium consisting of secretory, ciliated, and basal cells. Immunohistochemistry, using antibodies to mucin and lysozyme, and lectin histochemistry revealed both mucous and serous secretory cells in the epithelium. Cell proliferation was studied in situ using antibodies to proliferating cell nuclear antigen (PCNA) and Ki-67. Whereas at the time of explantation the proliferation was low (2.5±1.7% of the epithelial cells were PCNA-positive, 1.7±0.6 were Ki-67-positive), at 24 h of cultivation, 30.4±5.1% or 25.2±4.9% of the epithelial cells were labeled with antibodies to PCNA or Ki-67. After 7 days, the number of dividing cells was low again. The results show that the organ-culture system of human respiratory surface epithelium produces a differentiated epithelium that is useful in the study of secretory processes, differentiation, and proliferation.
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  • 69
    ISSN: 1432-0878
    Keywords: Key words Mammary gland ; Type XIV collagen ; Undulin ; Fibroblast ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Immunolocalisation of type XIV collagen/undulin in the human mammary gland revealed greater deposition in the interlobular stroma than in the intralobular stroma. The interlobular stroma is located between the breast lobules and their associated intralobular stroma. Fibroblasts isolated from the interlobular stroma synthesised 3- to 5-fold more type XIV collagen/undulin than intralobular fibroblasts, but synthesised type I and type IV collagens in similar amounts. The differential expression of type XIV collagen/undulin was maintained with passage in culture. The results suggest a role for type XIV collagen/undulin in stabilising dense collagen fibrils. The maintenance of two types of structurally distinct stromas may be important during developmental processes in the mammary gland.
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  • 70
    ISSN: 1432-0878
    Keywords: Key words TGF-β ; TGF-β receptors ; Vascular endothelium ; Heart ; Vascular disorders ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Endoglin is a component of the receptor complex for transforming growth factor (TGF)-β1 and TGF-β3. We analysed its expression by immunohistochemistry in human embryos at 4–8 weeks of gestation and in hearts ranging from 4–13 weeks old. We compared endoglin distribution with that of TGF-β receptors type I (TβR-I), type II (TβR-II) and betaglycan. Endoglin was found on endothelial cells in all tissues examined, consistent with its expression in adult blood vessels. TβR-I, TβR-II and betaglycan were observed on most cell types and had an overall similar pattern of distribution. Endoglin was detected on the endocardium as early as 4 weeks, but was absent from myocardium. It was present at high levels on the endocardial cushion tissue mesenchyme from 5–8 weeks’ gestation, during heart septation and valve formation, and subsequently decreased as the valves matured. Endoglin expression in heart extracts was confirmed by Western blot analysis. TβR-I, TβR-II and betaglycan were mostly found on cardiac myocytes, but were detectable at low levels on endocardium. They were expressed transiently on cushion mesenchyme, albeit at much lower levels than endoglin. All four components of the TGF-β receptor complex were detected by RT-PCR in embryonic heart. Thus transient up-regulation of the components of the TGF-β receptor complex, and particulartly of endoglin, is associated with heart septation and valve formation during early human development.
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  • 71
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    Cell & tissue research 293 (1998), S. 285-291 
    ISSN: 1432-0878
    Keywords: Key words Muscle ; Masseter ; Biceps ; P blood group ; CD77 ; Galactose ; Cat ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  There is evidence that glycans carrying terminal galactose residues are differently expressed in the sarcoplasm of different muscle fiber types. In this study monoclonal antibodies directed against P blood group antigens Pk: Galα1–4Galβ1–4Glcβ- and P1: Galα1–4Galβ1–4GlcNAcβ- were used to detect terminal α-galactosylated glycoconjugates on muscle proteins. Electrotransfer of proteins, extracted from human masseter and biceps muscles, to nitrocellulose after polyacrylamide gel electrophoresis (PAGE) and incubation with anti-Pk (CD77) consistently showed two bands with apparent molecular weights of 66 kDa and 64 kDa. In fresh frozen muscle sections from some humans there was endothelial reaction with anti-CD77 in capillaries, venules and veins but not in arterioles and arteries. In muscle samples from other humans there was no staining of endothelial cells. Formalin-fixed human muscle displayed a CD77 reaction with highest accumulation of reaction product at the periphery of the fibers. This may be explained by the presence of Pk glycoconjugates on intermediate filaments in muscle fibers. In preparations of cat masseter muscle proteins the antibodies against P1Pk antigens reacted with a 170 kDa and a 55 kDa band while in preparations of cat biceps brachii only a 55 kDa band was reactive. The specificities of the antibodies were investigated by fluorescence-activated cell sorter (FACS), α- and β-galactosidase digestion and inhibitory sugars. This study indicates that glycans carrying Galα1–4Galβ1- epitopes are expressed on myofibrillar associated proteins.
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  • 72
    ISSN: 1432-0878
    Keywords: Key words Nose ; Endothelin ; Respiratory epithelium ; Metaplastic epithelium ; Nasal blood flow ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The presence and site of production of endothelin-1 (ET-1) was investigated in biopsies obtained from the nasal mucosa of 10 healthy human subjects and 10 patients affected by chronic rhinitis. The presence and localization of receptors for ET-1 was also investigated. Bioptic fragments were examined by scanning electron microscopy. ET-1 was present in the vessels and in the respiratory epithelium of normal subjects, whereas in patients affected by epithelial metaplasia induced by chronic rhinitis, it was absent in the metaplastic epithelium and present in the endothelium and vascular wall. Receptors for ET (A- and B-receptor subtypes) were localized in the vessels of the nasal mucosa, both in normal and in pathological subjects. In particular, A-receptors were identified in the vascular wall, whereas B-receptors were mainly distributed in the endothelium. We suggest that ET-1 is involved in the homeostasis of nasal blood flow (shunting the blood toward the deep cavernous plexus and inducing mucosal swelling) by an autocrine and/or paracrine mechanism. Normal epithelium seems to be important in this mechanism, since it is able to produce ET. However, when pathologic conditions induce squamous or cuboidal metaplasia, the epithelium is no longer able to play this role.
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  • 73
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    Cell & tissue research 292 (1998), S. 91-99 
    ISSN: 1432-0878
    Keywords: Key words Argyrophil cells ; Chronic gastritis ; Gastrinoma/Zollinger-Ellison syndrome ; Hypergastrinaemia ; Neuroendocrine complexes ; Pernicious anaemia ; Proton pump inhibitors ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Hypergastrinaemia-associated changes of non-antral argyrophil cells in man are of increasing interest, because of the development of potent inhibitors of gastric acid secretion. Using an antibody against chromogranin A, we identified micronodular endocrine cell hyperplasia of the oxyntic mucosa in gastric biopsy specimens of patients with hypergastrinaemia of different backgrounds. Consecutive ultrathin sections were examined at the electron-microscopical level. Endocrine cell types within the (extraepithelial) micronodules closely resembled those in the adjacent mucosa. Micronodules were classified into two groups. The first group was composed of endocrine cells only and predominated in patients with drug-induced hypergastrinaemia and/or chronic gastritis, and in a gastrinoma/MEN I patient. The second group represented “neuroendocrine complexes”, showing a close intermingling of non-myelinated nerve fibres with endocrine cells, and was found predominantly in pernicious anaemia. Micronodular argyrophil cell growth in man is therefore heterogeneous and depends on the background of the hypergastrinaemia.
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  • 74
    ISSN: 1432-0878
    Keywords: Key words Reconstructed epidermis ; Differentiation ; Proliferation ; Migration ; Loricrin ; Involucrin ; SPRR ; EGF ; TGFα ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Epidermis reconstructed on de-epidermized dermis was used to investigate the effects of growth factors and culture temperature on epidermal morphogenesis and the expression of cornified envelope precursors. Cultures grown at 33°C or 37°C in the absence or presence of transforming growth factor alpha (TGFα), keratinocyte growth factor (KGF), basic fibroblast growth factor (bFGF), or insulin-like growth factor (IGF) show a similar morphology to that of native epidermis. Loricrin and SPRR2 are expressed in the stratum granulosum and SPRR3 is absent. Cultures grown in epidermal growth factor (EGF)-supplemented medium at 37°C have a normal morphology, whereas cultures grown at 33°C have a disorganized basal layer, no stratum granulosum, and nuclei are present in the stratum corneum. Loricrin is absent, and SPRR2 and SPRR3 expression extend into the spinous layers. Irrespective of the culture condition used, involucrin is aberrantly expressed in all suprabasal layers. EGF stimulated keratinocyte proliferation and migration to a greater degree than TGFα. Epidermis reconstructed on fibroblast-populated collagen gels at 33°C led to the same disturbances in keratinocyte differentiation as seen in cultures grown on de-epidermized dermis at 33°C in the presence of EGF, whereas parallel cultures grown at 37°C have a similar morphology to that of native epidermis.
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  • 75
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    International journal of salt lake research 7 (1998), S. 235-260 
    ISSN: 1573-8590
    Keywords: groundwater ; Merseburg-Ost ; mine ; meromixis ; numerical model ; monimolimnion ; opencast ; physical limnology ; salinity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geography
    Notes: Abstract Salt-laden groundwater inflows have formed monimolimnia in developing lakes of the abandoned opencast lignite mine Merseburg-Ost, Central Germany. Water-levels are still rising. To predict the future status of the lakes, two numerical models were developed. One was designed to predict groundwater flows, the second included limnophysical processes. The models were run in parallel and data exchange between them was performed every 1 to 10 years of model time. A trial of a 100-years prognosis was done. The results of surface salinity, mixing depth, and salinity profile are given. A sensitivity analysis yielded indications for the accuracy of the prediction.
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  • 76
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    International journal of salt lake research 7 (1998), S. 261-274 
    ISSN: 1573-8590
    Keywords: cyanobacteria ; lake management ; Mono Lake ; nitrogen fixation ; salinity ; saline lakes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geography
    Notes: Abstract Mono Lake is a hypersaline alkaline lake in the high altitude Great Basin desert of eastern California. Algal productivity of the lake is nitrogen-limited, and a contributing source is derived from benthic nitrogen fixation. Lake level and salinity have fluctuated with natural climatic variations but have also been affected by the diversion of tributary streams. This research examines the influence of varied salinity and lake level on the potential for benthic nitrogen fixation in Mono Lake. A sediment-surface microbial mat community was exposed directly, and in acclimated cultures, to a range of Mono Lake salinities under anaerobic incubations and the activity of nitrogenase assayed by acetylene reduction. Activity was stimulated in light, but also occurred in darkness. Over an experimental salinity range from 50 to 150 g L−1 TDS, nitrogenase activity was reduced by 90 per cent, with the activity persisting at the highest salinity being attributable to dark fixation alone. Between a salinity of 50 g L−1, occurring in Mono Lake over 50 years ago, and 100 g L−1, nitrogenase activity was reduced by nearly half. Changes in the area of the littoral zone at varied lake levels also affect the total amount of potential benthic nitrogen fixation in the lake. An accounting of yearly inputs of nitrogen to Mono Lake suggests N2-fixation could contribute as much as 76–81 percent of the total. Inhibition of nitrogen fixation rates by increased salinity could limit the long-term nutrient supply and benthic primary productivity of this ecosystem.
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  • 77
    ISSN: 1573-675X
    Keywords: Androgen ; apoptosis ; heat stress ; hormone ; temperature ; testis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Previous studies have demonstrated that testicular germ cell apoptosis can be induced both by heat stress and by withdrawal of androgens and gonadotrophins. To investigate whether heat-induced germ cell apoptosis occurs independently of the altered levels of hormones that occur with heat exposure, mouse testicular apoptosis was studied using an in vitro system with controlled levels of testosterone, FSH and LH. It was observed that cells underwent apoptosis sooner in the absence of hormones at the same temperature. Apoptosis also occurred earlier at abdominal temperature compared to scrotal temperature with the same hormonal levels. No somatic tissues studied underwent apoptosis at 37°C under the same culture conditions. These results suggest that heat stress may independently activate an apoptotic pathway in the testis, and that hormone deprivation may induce apoptosis via a separate mechanism.
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  • 78
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    International journal of salt lake research 7 (1998), S. 153-170 
    ISSN: 1573-8590
    Keywords: confounding parameters ; nutrients ; salinity ; survey design
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geography
    Notes: Abstract In order to relate macroinvertebrate fauna to electrical conductivity (EC), a survey of selected orders of macroinvertebrates was conducted in the Glenelg, Hopkins, Merri and Barwon River Catchments in south-west Victoria, Australia. The survey consisted of sampling 21 paired sites at confluences of streams. There was little difference between the in-stream habitat but there was a range of differences in EC between each of the paired sites. The dissimilarity of macroinvertebrate fauna between the paired sites was compared to the proportional difference in EC and other water quality variables (dissolved oxygen, water temperature, pH, NO2−N, NO3−N, PO4 and turbidity). This allowed the elimination of EC being confounded with geographical scale parameters and took into account other water quality parameters that may be confounded with EC. A positive relationship was found between the dissimilarity of the fauna at each of the two paired sites and the proportional difference in EC. No such relationship was found for any other measured water quality variable. The results of this survey show that macroinvertebrate community structure is associated with EC of the river systems investigated after eliminating large scale geographical patterns, in-stream habitat and the measured water quality parameters.
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  • 79
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    International journal of salt lake research 7 (1998), S. 261-274 
    ISSN: 1573-8590
    Keywords: cyanobacteria ; lake management ; Mono Lake〉/kwd〉 ; nitrogen fixation ; salinity ; saline lakes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geography
    Notes: Abstract Mono Lake is a hypersaline alkaline lake in the high altitude Great Basin desert of eastern California. Algal productivity of the lake is nitrogen-limited, and a contributing source is derived from benthic nitrogen fixation. Lake level and salinity have fluctuated with natural climatic variations but have also been affected by the diversion of tributary streams. This research examines the influence of varied salinity and lake level on the potential for benthic nitrogen fixation in Mono Lake. A sediment-surface microbial mat community was exposed directly, and in acclimated cultures, to a range of Mono Lake salinities under anaerobic incubations and the activity of nitrogenase assayed by acetylene reduction. Activity was stimulated in light, but also occurred in darkness. Over an experimental salinity range from 50 to 150 g L-1 TDS, nitrogenase activity was reduced by 90 per crnt, with the activity persisting at the highest salinity being attributable to dark fixation alone. Between a salinity of 50 g L-1, occurring in Mono Lake over 50 years ago, and 100 g L-1, nitrogenase activity was reduced by nearly half. Changes in the area of the littoral zone at varied lake levels also affect the total amount of potential benthic nitrogen fixation in the lake. An accounting of yearly inputs of nitrogen to Mono Lake suggests N2-fixation could contribute as much as 76–81 percent of the total. Inhibition of nitrogen fixation rates by increased salinity could limit the long-term nutrient supply and benthic primary productivity of this ecosystem.
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  • 80
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    International journal of salt lake research 7 (1998), S. 235-260 
    ISSN: 1573-8590
    Keywords: groundwater ; Merseburg-Ost ; mine ; meromixis ; numerical model ; monimolimnion ; opencast ; physical limnology ; salinity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geography
    Notes: Abstract Salt-laden groundwater inflows have formed monimolimnia in developing lakes of the abandoned opencast lignite mine Merseburg-Ost, Central Germany. Water-levels are still rising. To predict the future status of the lakes, two numerical models were developed. One was designed to predict groundwater flows, the second included limnophysical processes. The models were run in parallel and data exchange between them was performed every 1 to 10 years of model time. A trial of a 100-years prognosis was done. The results of surface salinity, mixing depth, and salinity profile are given. A sensitivity analysis yielded indications for the accuracy of the prediction.
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  • 81
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    International journal of salt lake research 7 (1998), S. 285-299 
    ISSN: 1573-8590
    Keywords: nutrients ; salinity ; water quality
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geography
    Notes: Abstract The effect of saline lake water disposal from the Lough Calvert Drainage Scheme on water quality of the Barwon River, in south west Victoria, Australia, was investigated. The Scheme is used to regulate the levels of several saline lakes outside the Barwon's catchment. This study found that the disposal of saline lake water was associated with increased total phosphorus, PO4, TKN, suspended solids, electrical conductivity and stream discharge and lower NOx in the Barwon River. Thus, when disposing of saline water, factors other than salinity should be considered in order to prevent an impact on the aquatic environment. At present this is generally not done. The results are discussed with reference to the effects of saline water disposal on aquatic biota and how the effect of saline lake water disposal on water quality may differ from the disposal of saline groundwater.
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  • 82
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    International journal of salt lake research 7 (1998), S. 285-300 
    ISSN: 1573-8590
    Keywords: nutrients ; salinity ; water quality
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geography
    Notes: Abstract The effect of saline lake water disposal from the Lough Calvert Drainage Scheme on water quality of the Barwon River, in south west Victoria, Australia, was investigated. The Scheme is used to regulate the levels of several saline lakes outside the Barwon's catchment. This study found that the disposal of saline lake water was associated with increased total phosphorus, PO4, TKN, suspended solids, electrical conductivity and stream discharge and lower NOx in the Barwon River. Thus, when disposing of saline water, factors other than salinity should be considered in order to prevent an impact on the aquatic environment. At present this is generally not done. The results are discussed with reference to the effects of saline water disposal on aquatic biota and how the effect of saline lake water disposal on water quality may differ from the disposal of saline groundwater.
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  • 83
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    International journal of salt lake research 7 (1998), S. 153-170 
    ISSN: 1573-8590
    Keywords: confounding parameters ; nutrients ; salinity ; survey design
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geography
    Notes: Abstract In order to relate macroinvertebrate fauna to electrical conductivity (EC), a survey of selected orders of macroinvertebrates was conducted in the Glenelg, Hopkins, Merri and Barwon River Catchments in south-west Victoria, Australia. The survey consisted of sampling 21 paired sites at confluences of streams. There was little difference between the in-stream habitat but there was a range of differences in EC between each of the paired sites. The dissimilarity of macroinvertebrate fauna between the paired sites was compared to the proportional difference in EC and other water quality variables (dissolved oxygen, water temperature, pH, NO2-N, NO3-N, PO4 and turbidity). This allowed the elimination of EC being confounded with geographical scale parameters and took into account other water quality parameters that may be confounded with EC. A positive relationship was found between the dissimilarity of the fauna at each of the two paired sites and the proportional difference in EC. No such relationship was found for any other measured water quality variable. The results of this survey show that macroinvertebrate community structure is associated with EC of the river systems investigated after eliminating large scale geographical patterns, in-stream habitat and the measured water quality parameters.
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  • 84
    ISSN: 1573-2614
    Keywords: heparin ; anticoagulation ; cardiopulmonary bypass ; temperature ; normothermia ; hypothermia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Computer Science , Medicine
    Notes: Abstract Objective. With the practice of warm cardiopulmonary bypass (CPB) at our institution we have observed an apparent increase in heparin requirements. CPB temperature predictability affects pharmacokinetics and differences in drug metabolism can be expected. We hypothesized that heparin requirements would increase with increasing CPB temperature. Methods. Following Institutional Review Board approval, we reviewed the charts of 354 patients undergoing primary coronary artery bypass graft surgery. We recorded patient demographic data, CPB duration, heparin requirements, and temperature during CPB. CPB was conducted between 24 °C and 37 °C. The Spearman's correlation coefficient, Pearson chi-square, and rank-sum tests were used for data analysis. Results. Core temperature during CPB correlated with heparin requirements (r = 0.13, p 〈 0.02). However, CPB duration was shorter in warm patients than in cold patients (r = −0.455, p 〈 0.0001). Additional heparin requirements adjusted for duration of CPB (units/minute) were also significantly greater in the warm group (p = 0.018). Conclusions. Maintenance of adequate heparin anticoagulation during CPB is clinically important. Warm CPB patients required more heparin per minute than those undergoing cold CPB. More frequent assessment of anticoagulation and administration of additional heparin should be considered in patients undergoing warm CPB.
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  • 85
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    Bulletin of experimental biology and medicine 126 (1998), S. 1140-1143 
    ISSN: 1573-8221
    Keywords: liver ; platidiam ; pharmorubicine ; CCl 4-induced hepatitis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Experiments performed on rats have shown that a single intravenous injection of the antitumor preparations platidiam and pharmorubicine in the maximum tolerated dose cause morphological changes in the liver which are preserved for 3 months after platidiam and for 6 moths after pharmorubicine. Instability of reparative processes in the liver was revealed when the rats were poisoned with CCl4, 1, 3, and 6 months after administration of the cytostatics.
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  • 86
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    Bulletin of experimental biology and medicine 126 (1998), S. 734-736 
    ISSN: 1573-8221
    Keywords: liver ; endothelial cells of sinusoids ; long-term compression syndrome
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Electron microscopic and morphometric studies showed reduction in the intensity of plastic processes, impaired mitochondrial function, and increased proportion of secondary lysososomes in endothelial cells of liver sinusoids in rats with a moderate long-term compression syndrome. These processes were pronounced in dextran-treated rats, which may be due to impaired trophics of hepatocytes.
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  • 87
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    Bulletin of experimental biology and medicine 126 (1998), S. 731-733 
    ISSN: 1573-8221
    Keywords: liver ; hepatocyte ; physical load ; clasmatosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract In rats, running of the maximum intensity caused death of some hepatocytes, an increase in the number of phagosomes in Kuppfer cells, and the emergence of connective tissue fibers in the space of Disse. Ultrastructural investigation of hepatocytes showed delayed release of bile products into bile capillaries, decrease in glycogen content, increase in the number of mitochondria (many of them were divided by the cristae), and irregular distribution of ribosomes in the rough endoplasmic reticulum. Accumulation of erythrocytes in the sinusoids, fragments of dead hepatocytes, Kuppfer cells with numerous phagosomes, and connective tissue fibers in the space of Disse were observed in rat liver after exhausting swimming. Study of hepatocyte ultrastructure revealed intense protein synthesis (as evidenced by increased number of ribosomes and unchanged mitochondria and cisternae of the rough endoplasmic reticulum), separation of cytoplasmic fragments with ribosomes into sinusoids, absence of glycogen, and lipid accumulation.
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  • 88
    ISSN: 1573-8221
    Keywords: regeneration ; lymphoid tissue reaction ; liver ; pancreas
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Resection of the liver and pancreas in birds involves a decrease in mitotic activity of follicular lymphoid cells in the bursa of Fabricius (3.8 and 2.6 times on days 10 and 20 postoperation, respectively) and in the spleen (1.5–2 times 1–30 days postoperation) in comparison with the white pulp and splenic follicles volume. In frogs, unilateral nephrectomy involves devastation of the spleen (its red and white pulp), which does not decrease by day 8 postoperation.
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  • 89
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    European journal of nutrition 36 (1997), S. 23-27 
    ISSN: 1436-6215
    Keywords: Vitamin E ; Fleisch ; Fettgewebe ; Leber ; Eigelb ; Vitamin E ; meat ; adipose tissue ; liver ; egg yolk
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition , Medicine
    Description / Table of Contents: Summary The α-tocopherol content of different meat cuts was examined. Chicken thigh had the highest vitamin E content, followed by chicken breast and pork shoulder (p〈0.05). The lowest concentrations were found in longissimus dorsi muscle from pork, beef, veal and in beef shoulder. Considering the average daily lean meat consumption (105 g) in Switzerland, recommendation for daily vitamin E intake was met to 3 %. Supplementation of 200 mg α-tocopherol acetate/kg feed to pigs and laying hens significantly increased the α-tocopherol content in all examined products. The α-tocopherol accumulation differed according to the following ranking: egg yolk 〉 liver 〉 adipose tissue 〉 musculus longissimus dorsi. The α-tocopherol:energy ratios were 28.8, 7.3, 0.9 and 1.2 mg/MJ for egg yolk, liver, adipose tissue and longissimus dorsi muscle of the vitamin E supplemented groups, respectively. The results showed that meat, with the exception of chicken thigh, is not an important supplier of vitamin E, not even from animals fed a vitamin E enriched diet. Egg yolk became a good source of vitamin E for human nutrition by dietary modification.
    Notes: Zusammenfassung In der vorliegenden Studie wurde der α-Tocopherolgehalt verschiedener Fleischstücke untersucht. Hähnchenschenkel hatte den höchsten α-Tocopherolgehalt, gefolgt von Hähnchenbrust und Schweineschulter (p〈0.05). Die niedrigsten Konzentrationen wurden im Musculus longissimus dorsi vom Schwein, Rind, Kalb und in der Rindsschulter nachgewiesen. Mit dem durchschnittlichen, täglichen Verzehr an magerem Fleisch (105 g) in der Schweiz wurden die Empfehlungen für die tägliche Vitamin E-Zufuhr zu 3 % gedeckt. Die Supplementierung des Schweine- und Legehennenfutters mit 200 mg α-Tocopherolacetat/kg führte zu einem signifikanten Anstieg des α-Tocopherolgehaltes in allen untersuchten Produkten. Die α-Tocopherolakkumulierung unterschied sich gemäß folgender Rangordnung: Eigelb 〉 Leber 〉 Fettgewebe 〉Musculus longissimus dorsi. Die Nährstoffdichten betrugen 28.8, 7.3, 0.9 und 1.2 mg α-Tocopherol/MJ für Eigelb, Leber, Fettgewebe und Musculus longissimus dorsi der jeweiligen mit Vitamin E supplementierten Gruppe. Diese Ergebnisse zeigen, daß Fleisch, mit Ausnahme des Hähnchenschenkels, von Tieren mit supplementierten Diäten kein bedeutender Vitamin E-Lieferant ist. Hingegen wurde Eigelb durch fütterungsbedingte Modifikation zu einer guten Vitamin E-Quelle.
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  • 90
    ISSN: 1436-6215
    Keywords: Riboflavin ; Milch ; Leber ; Restkörper ; Laktation ; Ratte ; Riboflavin ; milk ; liver ; carcass ; lactation ; rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition , Medicine
    Description / Table of Contents: Summary The present study investigated the effect of various dietary riboflavin supplementations (0 to 4 000 mg/kg) during lactation on riboflavin concentrations of liver, carcass (bled body without intestine and liver), and milk in the rat. The experiment was conducted until the 14th day of lactation; milk samples were drawn on the 7th and 13th day of lactation. Riboflavin concentrations of milk raised continuously with increasing riboflavin supplementation; in the range between 0 and 10 mg/kg riboflavin supplementation, there was a linear relationship, and in the range between 12 and 4 000 mg/kg there was a logarithmic relationship between riboflavin supplementation and riboflavin concentration in the milk. Maximum riboflavin concentration of milk obtained by supplementation with 4 000 mg/kg was twelve-fold higher than without riboflavin supplementation. For riboflavin supplementation up to 12 mg/kg, riboflavin concentrations in milk on the 7th day of lactation and that on the 13th day of lactation were not different. In contrast, in rats fed diets with higher riboflavin supplementation, riboflavin concentrations were higher by 25 % in average in milk on the 13th day of lactation than in milk on the 7th day of lactation. Contrary to the milk, riboflavin concentrations in liver and carcass exhibited a saturation, which was achieved at a supplementation of 6 mg/kg (liver) and 10 mg/kg (carcass), respectively. Maximum riboflavin concentrations obtained at a supplementation of 4 000 mg/kg were 1.9- and 2.3-fold higher for liver and carcass, respectively, than concentrations obtained without riboflavin supplementation. The dose-response relationship using riboflavin concentrations of liver and carcass as response factors indicates a riboflavin requirement of 8 to 9 mg/kg for lactating rats fed a semisynthetic diet with 17.4 MJ ME/kg dry matter and 20.8 % protein in dry matter.
    Notes: Zusammenfassung In der vorliegenden Arbeit wurde der Einfluß unterschiedlicher Riboflavinzulagen zum Futter (0 bis 4 000 mg/kg) während der Laktation auf die Riboflavinkonzentrationen in Leber, Restkörper (ausgebluteter Gesamtkörper ohne Magen-Darm-Trakt und Leber) und Milch von Ratten untersucht. Der Versuch dauerte bis zum 14. Laktationstag; Milchproben wurden am 7. und am 13. Laktationstag gewonnen. Die Riboflavinkonzentration der Milch erhöhte sich mit steigender Zulage stetig, wobei im Bereich zwischen 0 und 10 mg Riboflavinzulage/kg Futter eine lineare und im Bereich zwischen 12 und 4 000 mg/kg eine logarithmische Funktion vorlag. Die maximale Riboflavinkonzentration in der Milch bei einer Zulage von 4 000 mg/kg war dabei etwa zwölfmal so hoch wie bei fehlender Zulage. Bei Riboflavinzulagen bis 12 mg/kg unterschieden sich die Riboflavinkonzentrationen der Milch am 7. und 13. Laktationstag nicht. Bei den höheren Zulagen waren die Konzentrationen der Milch am 13. Laktationstag im Mittel um 25 % höher als am 7. Laktationstag. Im Gegensatz zur Milch zeigte sich in Leber und Restkörper eine Sättigung der Riboflavinkonzentrationen, die bei einer Riboflavinzulage von 6 mg/kg (Leber) bzw. 10 mg/kg (Restkörper) erreicht war. Die maximalen Riboflavinkonzentrationen bei Zulagen von 4 000 mg/kg waren dabei 1,9 (Leber) bzw. 2,3 (Restkörper) mal so hoch wie bei fehlender Riboflavinzulage. Diese Befunde sprechen für eine ausgeprägte homöostatische Kontrolle der Riboflavinkonzentrationen im Organismus. Anhand von Dosis-Wirkungsbeziehungen mit den Riboflavinkonzentrationen in Leber und Restkörper als Wirkungskriterien leitete sich bei Verwendung des halbsynthetischen Futters (17,4 MJ ME/kg Trockenmasse (T), 20,8 % Rohprotein in T) ein Riboflavinbedarf von 8 bis 9 mg/kg Futter für die laktierende Ratte ab.
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  • 91
    ISSN: 1436-6215
    Keywords: Environmental temperature ; energy expenditure ; ouabain-sensitive respiration ; duodenal mucosa ; liver ; rats ; Umgebungstemperatur ; Energieumsatz ; Quabain-sensitive Respiration ; Duodenalmukosa ; Leber ; Ratten
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition , Medicine
    Description / Table of Contents: Zusammenfassung Die Wirkung der Umgebungstemperatur (18°C oder 28°C) und des Fasergehalts in der Diät (g je kg Trockensubstanz (TS) niedrig - 68, mittel - 110, hoch - 157) oder des Proteingehalts (g je kg TS niedrig - 91, mittel- 171, hoch - 262) auf den Verdauungstrakt, die Darmmasse, den Energieumsatz und auf die mit der Na+, K+-ATPase-Aktivität zusammenhängenden Respiration von Duodenalmukosa und Leber wurde bei 72 Wistar-Ratten in wiederholten Experimenten untersucht. Der Gesamte und Quabain-sensitive (ein Maß der Na+, K+-ATPase Aktivität) O2-Verbrauch der Gewebe wurde in vitro polarographisch ermittelt (YSI-biologische Sauerstoff-Erfassung nach dem Clark-Meßprinzip). Die Wärmeproduktion (WP) intakter Tiere wurde über Respirationskammern mit offenem Gasaustausch erfaßt. Die bei 18°C gehaltenen Ratten wiesen im Vergleich zu 28°C eine höhere Darmmasse auf. Die Masse an leerem Dünndarm, Caecum und Colon stieg mit ansteigendem Fasergehalt in der Diät (P〈0.05). Die WP als Korrelat der umsetzbaren Energie war nur im 1. Experiment höher (P〈0.05) bei 18°C als bei 28°C. Bei niedriger Proteinstufe war die WP signifikant höher (P 0.05) als bei den anderen Stufen. Verglichen mit 28°C erzeugte 18°C einen ansteigenden Gesamt- und Quabain-sensitiven O2-Verbrauch in der Duodenalmukosa. Die Leber reagierte nicht auf Temperaturunterschiede. Jedoch war ihr Quabain-sensitiver O2-Verbrauch bei niedrigem Proteingehalt in der Nahrung höher (P〈0.05) als bei den anderen Varianten. Bei niedrigem Fasergehalt war der gesamte und Quabain-sensitive O2-Verbrauch der Duodenalmukosa höher als bei den anderen Fasergehaltsvarianten. Die In-vitro-Ergebnisse stimmten mit der WP und dem O2-Verbrauch intakter Tiere überein.
    Notes: Summary Seventy two Wistar rats were used in two repeat studies to investigate the effect of environmental temperature (18°C or 28°C) and increasing levels of dietary fibre (low, 68 g/kg DM; medium 110 g/kg DM; high, 157 g/kg DM) or protein (low, 91 g/kg DM; medium, 171 g/kg DM; high, 262 g/kg DM) on digestive tract, visceral organ size, energy metabolism, and respiration attributable to Na+,K+-ATPase activity in duodenal mucosa and liver. Total and ouabain-sensitive (a measure of Na+,K+-ATPase activity) O2 consumptionin vitro of tissues were measured polarographically using a Clark-style YSI biological O2 monitor. Whole body heat production (in vivo) was measured using open-circuit respiration chambers. The weight of the visceral organs was higher in rats housed at 18°C than at 28°C. The empty weight of the small intestine, caecum, and colon increased as the level of dietary fibre increased (P 0.05). Heat production as a proportion of metabolizable energy was higher (P〈0.05) at 18°C than at 28°C in the first experiment but this difference was not significant in the second experiment. Rats fed the low protein diet had significantly higher (P〉0.05) heat production than those fed medium or high protein diets. Compared to 28°C, environmental temperature of 18°C caused an increased total and ouabain-sensitive O2 consumption in duodenal mucosa. There was no significant effect of environmental temperature on total and ouabain-sensitive O2 consumption in the liver. However, ouabain-sensitive O2 consumption in liver was significantly higher (P 0.05) when rats were fed a low protein diet compared to the medium or high protein diet. Total and ouabain-sensitive O2 consumption increased in duodenal mucosa of rats fed low level of dietary fibre compared to the medium or high dietary fibre diets. Thein vitro results corresponded with the whole animal energy expenditure and O2 consumptionin vivo.
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  • 92
    ISSN: 1573-0832
    Keywords: Anthracnose ; guava ; Colletotrichum glososporioides ; temperature ; free moisture ; inoculum density
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The influence of environmental factors (temperature and humidity), inoculum density on infection by Colletotrichum glososporioides and development of anthracnose lesions were determined on uninjured, sand-injured and punctured fruits. The optical temperature for severe infection was 30 °C, whereas the disease incidence was less at 20 and 35 °C. Inoculated guavas that received 1–60 h of continuous free moisture developed lesions, but the disease was minimal (0–7%) after 1–6 h free moisture. Infection rates of uninjured, sand-injured and punctured fruits receiving 60 h of free moisture were 34, 70 and 100%, respectively. Disease incidence increased as inoculum density increased from 101 to 106 conidia/ml. In field conditions, the development of anthracnose lesions was greater on punctured guavas than on uninjured or sand-injured ones, in both rainy and winter seasons. In general, the number of lesions was highest in sand-injured fruits, followed by punctured and uninjured fruits. In rainy season the number of lesions on injured and uninjured fruits was greater than similarly treated guavas in winter.
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  • 93
    ISSN: 1573-0832
    Keywords: Antagonism ; biocontrol ; ginger ; postharvest rot ; Sclerotium rolfsii ; Trichoderma ; temperature
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Sclerotium rolfsii is a destructive soil-borne and postharvest plant pathogen. Use of the antagonistic fungus Trichoderma sp. has been earlier reported by us to successfully control this pathogen under postharvest conditions. In the present paper we report on the effects of temperature on the growth and biocontrol potential of Trichoderma sp. on S. rolfsii. Experimental results indicated that S. rolfsii and Trichoderma sp. have different temperature optima for growth: 30–35 °C for the pathogen and 25–30 °C for the antagonist. In dual culture, Trichoderma overgrew S. rolfsii at 25 °C and 30 °C, but at 35 °C and 37 °C, S. rolfsii overgrew the colony of Trichoderma. Trichoderma produced higher concentration of fungitoxic metabolites in broth culture at higher temperatures. In bioassays using ginger slices and whole rhizomes, it has been demonstrated that Trichoderma is not very effective in suppressing S. rolfsii at temperatures above 30 °C. In light of these results, possible mechanisms of biocontrol of S. rolfsii as a postharvest pathogen has been discussed. Storage temperature has been suggested as a critical factor in biocontrol of S. rolfsii.
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  • 94
    ISSN: 1432-1211
    Keywords: Key words HLA ; Microsatellite loci ; Microsatellite typing ; Human ; MHC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The human genome contains a large number of interspersed microsatellite repeats which exhibit a high degree of polymorphism and are inherited in a Mendelian fashion, making them extremely useful genetic markers. Several microsatellites have been described in the HLA region, but allele nomenclature, a set of broadly distributed controls, and typing methods have not been standardized, which has resulted in discrepant microsatellite data between laboratories. In this report we present a detailed protocol for genotyping microsatellites using a semi-automated fluorescence-based method. Twelve microsatellites within or near the major histocompatibility complex (MHC) were typed in the 10th International Histocompatibility Workshop homozygous typing cell lines (HTCs) and alleles were designated based on size. All loci were sequenced in two HTCs providing some information on the level of complexity of the repeat sequence. A comparison of allele size obtained by genotyping versus that obtained by direct sequencing showed minor discrepancies in some cases, but these were not unexpected given the technical differences in the methodologies. Fluorescence-based typing of microsatellites in the MHC described herein is highly efficient, accurate, and reproducible, and will allow comparison of results between laboratories.
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  • 95
    ISSN: 1432-1041
    Keywords: Key wordsProstaglandin E1 ; Carbonyl reductase; 13 ; 14-dihydro-15-keto-PGE1 ; 13 ; 14-dihydro-PGE1 ; human ; liver ; erythrocytes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Abstract Objective: The therapeutic response to PGE1 is highly variable, and a contribution by variable formation of its active tertiary metabolite PGE0 is in question. Hence, the objective of this study was to assess the person-to-person variation of the reduction of the inactive intermediate metabolite 15-KD PGE1 by human liver and human erythrocytes in forming the active metabolite PGE0. Methods: Source of enzyme was lysed erythrocytes from 29 donors, and a bank of 37 donor livers including specimens from 15 children. Tritium-labelled 13,14-dihydro-15-keto-prostaglandin E1 (15-KD PGE1) was used at low nanomolar concentrations and found to be converted almost exclusively to the more polar compound 13,14-dihydro-prostaglandin E1 (PGE0) by an NADPH-dependent carbonyl reductase. The identity of the product PGE0 was established by comparison of its chromatographic and mass spectral characteristics with authentic PGE0. Results: Lysed erythrocytes had readily measurable enzymatic activity; differences between the preparations from 29 subjects were very small with only a twofold range of variation. In contrast to lysed erythrocytes, intact erythrocytes did not catalyse the reaction so that the erythrocyte activity should be medically immaterial. 15-KD PGE1 15-ketoreductase activity of liver cytosol averaged 61.1 fmol · min−1 · mg−1 protein in preparations from 37 human livers. Individual activities varied over an almost tenfold range, with indications of a non-normal distribution. Kinetic studies of selected specimens showed substantially different Vmax values but indistinguishable k M values, suggesting that the individual variation in 15-KD PGE1 15-ketoreduction is the result of differences in enzyme concentration rather than of structural enzyme variations. The activity in 15 livers from children was significantly lower than in those from adults. Inhibition data suggest that both the liver and the erythrocyte enzymes belong to the class of carbonyl reductases. Conclusions: The variations in hepatic enzyme activity may be expected to affect the transformation of 15-KD PGE1 to the active metabolite PGE0 in vivo. The clinical significance remains to be explored.
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  • 96
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    Molecular and cellular biochemistry 167 (1997), S. 73-80 
    ISSN: 1573-4919
    Keywords: cadmium ; zinc ; liver ; flavokinase ; thiol group
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The effect of cadmium (Cd2+), mercury (Hg2+) and copper (Cu2+) was studied with partially purified flavokinase (ATP:riboflavin 5′-phosphotransferase EC 2.7.1.26) from rat liver. All the divalent heavy metal cations inhibited flavokinase activity in a concentration-dependent manner. The inhibitory effect of cadmium on the enzyme was completely reversed by increasing concentration, of Zinc (Zn2+) indicating a competition between Zn2+ and Cd2+ for binding with the enzyme. A competition between riboflavin and Cd2+ is also evident from the present investigation. These observations hint at the possibility that Zn2+ and Cd2+ probably compete for the same site on the enzyme where riboflavin binds. However, inhibition of flavokinase by Hg2+ could not be reversed by Zn2+. Our studies further reveal that hepatic flavokinase appears to contain an essential, accessible and functional thiol group(s) which is evident from a concentration dependent inhibition of activity by sulfhydryl reagent s like parachloromercuribenzoate (PCMB), 5,5′-dithiobis (2-nitrobenzoic acid)(DTNB), and N-ethylmaleimide (NEM). Inhibition of flavokinase by sulfhydryl reagents were protected, except in case of NEM inhibition, when the enzyme was incubated with thiol protectors like glutathione (GSH) and dithiothreitol (DTT). Furthermore, the enzyme could also be protected from the inhibitory effect of Cd2+ and Hg2+ by GSH and DTT suggesting that Cd2+ probably interacts with a reactive thiol group at or near the active site of enzyme in bringing about its inhibitory effect. (Mol Cell Biochem 167: 73-80, 1997)
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  • 97
    ISSN: 1573-4919
    Keywords: antioxidant enzymes ; sub-cellular organelles ; liver ; ischemia-reperfusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The activities of rat hepatic subcellular antioxidant enzymes were studied during hepatic ischemia/reperfusion. Ischemia was induced for 30 min (reversible ischemia) or 60 min (irreversible ischemia). Ischemia was followed by 2 or 24 h of reperfusion. Hepatocyte peroxisomal catalase enzyme activity decreased during 60 min of ischemia and declined further during reperfusion. Peroxisomes of normal density (d = 1.225 gram/ml) were observed in control tissues. However, 60 min of ischemia also produced a second peak of catalase specific activity in subcellular fractions corresponding to newly formed low density immature peroxisomes (d = 1.12 gram/ml). The second peak was also detectable after 30 min of ischemia followed by reperfusion for 2 or 24 h. Mitochondrial and microsomal fractions responded differently. MnSOD activity in mitochondria and microsomal fractions increased significantly (p 〈 0.05) after 30 min of ischemia, but decreased below control values following 60 min of ischemia and remained lower during reperfusion at 2 and 24 h in both organelle fractions. Conversely, mitochondrial and microsomal glutathione peroxidase (GPx) activity increased significantly (p 〈 0.001) after 60 min of ischemia and was sustained during 24 h of reperfusion. In the cytosolic fraction, a significant increase in CuZnSOD activity was noted following reperfusion in animals subjected to 30 min of ischemia, but 60 min of ischemia and 24 h of reperfusion resulted in decreased CuZnSOD activity. These studies suggest that the antioxidant enzymes of various subcellular compartments respond to ischemia/reperfusion in an organelle or compartment specific manner and that the regulation of antioxidant enzyme activity in peroxisomes may differ from that in mitochondria and microsomes. The compartmentalized changes in hepatic antioxidant enzyme activity may be crucial determinant of cell survival and function during ischemia/reperfusion. Finally, a progressive decline in the level of hepatic reduced glutathione (GSH) and concomitant increase in serum glutamate pyruvate transaminase (SGPT) activity also suggest that greater tissue damage and impairment of intracellular antioxidant activity occur with longer ischemia periods, and during reperfusion.
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  • 98
    ISSN: 1573-4919
    Keywords: cadmium ; glutathione S-transferase ; liver ; kidney
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Exposure of animals to cadmium (Cd) (25 mg kg-1 body wt day-1) for 10 weeks resulted in preferential accumulation of the metal in liver and kidney. Cd accumulation concomitantly increased zinc (Zn) concentration in both the organs. However, significant decrease in copper level was observed in liver, whereas kidney showed increase in copper (Cu) level. Cd exposure resulted in decreased total GST activity in liver (63%) and kidney (41%) as compared to control group monkeys on normal diet (group I). On isoelectric focusing (IFP) control liver GST segregated into thirteen isoenzymes, while in Cd-treated experimental animals (group II) liver GST resolved into nine isoenzymes. Similarly kidney GST from control animals separated into seven isoenzymes as compared to four isoenzymes from Cd-treated animals. Kinetic analysis showed that Cd exposure did not alter the affinity constant (Km) of GST for GSH and CDNB whereas maximal velocity (Vmax) for these substrates decreased as compared to controls in both the organs, indicating inhibition in GST synthesis by Cd. Cd resulted in a noncompetitive type of inhibition with respect to GSH in vitro. On isoelectric focussing GST of liver and kidney in group II resolved into nine and four isoenzymes as compared to thirteen and seven in group I, showing loss of four basic isoenzymes in case of liver and three isoenzymes in case of kidney. Monkey liver and kidney expressed all the three classes of GST isoenzymes i.e. α, µ and π, which were serologically identical to human α, µ and π GSTs. (Mol Cell Biochem 166: 55-63, 1997)
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  • 99
    ISSN: 1432-0878
    Keywords: Key words: Ageing ; Skin ; Proteinase ; Wound ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Despite the association of increasing age with chronic wound-healing disorders and an impaired rate of healing of acute cutaneous wounds, the role of matrix metalloproteinases (MMPs) is unknown. To determine the spatial and temporal patterns and activities of MMP-1, -2, -3 and -9, 132 healthy humans aged between 19 and 96 years underwent 4-mm punch biopsies followed by wound excision between day 1 and day 180 post-wounding. Wounds showed an age-related increase in MMP-2 and MMP-9 immunostaining from day 3; this was associated with degradation of gelatin as shown by zymograms and with increased proteinase activity as shown by azocoll assays. Distinct spatial localisations for each MMP were observed: MMP-2 was found in epidermal structures; MMP-9 was observed in inflammatory cells up to day 21; MMP-1 was localised to keratinocytes at the wound margin. Normal old skin showed pro-MMP-2 bands on zymography and increased MMP-2 immunostaining. These results indicate that: (1) intrinsic ageing is associated with the up-regulation of MMPs previously associated with chronic wound healing; (2) wound-tissue proteinases are essentially active up to day 21 postwounding; and (3) intrinsic ageing may predispose to tissue breakdown disorders because of MMP-2 up-regulation in normal skin.
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  • 100
    ISSN: 1432-0878
    Keywords: Key words: Fibroblast-specific antibody ; Fibroblast separation and elimination ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The unwelcome presence of fibroblasts in many cell cultures prevents the long term cultivation of various cell types and work with pure populations. Recently, we described a novel fibroblast-specific monoclonal antibody (MAb AS02) that recognises a membrane-bound antigen. We have now developed a method using the fibroblast-specific MAb AS02 immobilised on goat-anti-mouse-magnetic beads to separate contaminating fibroblasts. An endothelial cell line experimentally contaminated with 5%–50% fibroblasts was successfully purified. Additionally, an endothelial cell line with an initial fibroblast contamination of 1.5% was prepared. A proportion of each preparation was cultured with no separation step being performed, whereas the remainder was cultured after purification with MAb AS02 to exclude the presence of a minor number of fibroblasts (〈0.1%). The proportion of fibroblasts increased up to 38% in the fifth passage of culture without elimination of the low initial fibroblast contamination, whereas in the fraction with the separation step, no fibroblasts were detectable by flow cytometry, even after the fifth passage. We also used the antibody to detect the presence of naturally contaminating fibroblasts in thyrocyte cultures. After cultivation of thyrocyte cultures over five passages, the number of fibroblasts increased dramatically up to 50%–80% of the whole population. Subsequently, we successfully applied the method for complete elimination of naturally contaminating fibroblasts from freshly isolated thyrocyte cultures from enzymatically digested thyroid glands. Thus, MAb AS02 is a fibroblast-specific marker that is a useful tool for the detection and elimination of contaminating fibroblasts. The specificity of MAb AS02 permits the universal application of this antibody for human cell cultures of interest.
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