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  • 1
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Gamete Research 17 (1987), S. 15-20 
    ISSN: 0148-7280
    Keywords: nuclear transplantation ; electrofusion ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The present study was undertaken to determine the efficiency of HVJ treatment and electrofusion for pronuclear transplantation in the mouse. The output voltage and duration of the pulses were fixed to 200 μsec at 10 V or to 150 μsec at 15 V for electrofusion, because the maximum rates of blastomere fusion of 2-cell embryos and development of fused embryos in vitro were obtained under these conditions. Although the proportion of eggs with fused karyoplast (78%) and the fused eggs developed to morulae or blastocysts (67%) was significantly lower than those obtained after HVJ treatment (94% and 94%), the proportion of pregnant recipients and young obtained after treatment of fused eggs was not significantly different between these two procedures.It is advised that electrofusion can be used as a fusogenic procedure for pronuclear transplantation in the mouse in some cases where HVJ cannot be applied.
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  • 2
    ISSN: 0095-9898
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
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  • 3
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Journal of Morphology 217 (1993), S. 219-227 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The morphogenesis of glandular architecture of the three lobes of prostate gland of the guinea pig, lateral, dorsal, and coagulating gland was studied from 35 days gestation to 90 postnatal days. Epithelial ductal tubules of various lobes of the gland were microdissected after treatment by collagenase and displayed two dimensionally. The number of ductal tips was counted, and the volume of the ductal network was quantified using a graphic tablet. The results show that the growth and ductal morphogenesis fall into two phases: prenatal and postnatal. The first outgrowth of prostatic buds begins at 35 days gestation (gestational length is 65 days). Ductal growth and branching continues over the next 15-20 days and by 55 days gestation, approximately 60%, 79%, and 71% of the adult number of ductal tips of the lateral and dorsal lobes and coagulating gland respectively, are formed. The figures increase to 89%, 84%, and 106%, respectively, by birth. There is little increase in number of ductal tips thereafter. Postnatal growth is accomplished mainly by elongation of existing ductal network with a little additional branching but with an increase in size (volume) of the tubules. Canalization of ductal tubules occurs prenatally in all lobes but postnatal functional cytodifferentiation takes a slightly different pace among them. Ductal morphogenesis of the guinea pig prostate gland differs significantly in time-course from that of the mouse in which ductal development occurs mainly postnatally. © 1993 Wiley-Liss, Inc.
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  • 4
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Journal of Morphology 68 (1941), S. 507-517 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
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  • 5
    Electronic Resource
    Electronic Resource
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 46 (1991), S. 284-290 
    ISSN: 0730-2312
    Keywords: erythrocytes ; magnesium ; echinocyte ; calcium ; plasma membrane ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Intact human erythrocytes, initially depleted of Mg2+ by EDTA incubation in the presence of A23187, exhibit Mg2+-dependent phosphate production of around 1.5 mmol per liter cells · h, half-maximally activated at around 0.4 mM added free Mg2+. This appears to correspond to Mg2+-stimulated adenosine triphosphatase (Mg2+-ATPase) activity found in isolated membranes, which is known to have a similar activity and affinity for Mg2+. Vanadate (up to 100 μM) inhibited Mg2+-dependent phosphate production and ATP breakdown in intact cells. Over a similar concentration range vanadate (3-100 μM) transformed intact cells from normal discocytes to echinocytes within 4-8 h at 37°C, and more rapidly in Mg2+-depleted cells. The rate of Ca2+-induced echinocytosis was also enhanced in Mg2+-depleted cells. These results support previous studies in erythrocyte ghosts suggesting that vanadate-induced shape change is associated with inhibition of Mg2+-ATPase activity localized in the plasma membrane of the red blood cell.
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 30 (1995), S. 208-217 
    ISSN: 1059-910X
    Keywords: Superconductors ; Electron energy loss spectrometry ; Transmission electron microscope ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Electron energy loss spectrometry (EELS) with a cold field emission gun (cFEG) transmission electron microscope (TEM) is implemented to analyze the evolution of the electronic structure and dielectric function of oxide superconductors. The O-K core loss spectra of p-type doped oxide superconductors are analyzed in terms of holes formation on oxygen sites, while low loss spectra are analyzed for free carrier plasmas, other spectral excitations, and their crystallographic confinement.It is illustrated that the transmission EELS with a cFEG TEM very much complement soft X-ray absorption spectroscopy and optical spectroscopy, with the added advantages of high spatial resolution (∼1-100 nm), and is compatible with other analytical, diffraction, and imaging techniques, which are readily available in a cFEG TEM. © 1995 Wiley-Liss, Inc.
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  • 7
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 40 (1995), S. 228-232 
    ISSN: 1040-452X
    Keywords: Gossypol ; Sperm ; Acrosomal enzymes ; Rabbit ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The effect of gossypol on the activities of 10 acrosomal enzymes of the rabbit sperm was evaluated. Acrosin, Azocoll proteinase, neuraminidase, and arylsulfatase were significantly inhibited or completely inactivated by 12-76 μM gossypol. Hyaluronidase, β-glucuronidase, and acid phosphatase were inhibited only at a higher concentration of gossypol (380 μM). Phospholipase C, alkaline phosphatase, and β-N-Acetyl glucosaminidase were not inhibited even at 380 μM gossypol. Gossypol was found to be a noncompetitive inhibitor of arylsulfatase with a Ki of 120 μM. The inhibition was reversible and dose-dependent. As the acrosomal enzymes were more sensitive to the inhibition by gossypol compared to sperm enzymes involved in glycolysis or energy production, these assays may serve as a more reliable indicator for monitoring the occurence of gossypol-induced sterility. © 1995 Wiley-Liss, Inc.
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  • 8
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 25 (1990), S. 14-21 
    ISSN: 1040-452X
    Keywords: Sperm-specific ; Ldh-x ; Testis ; Gene expression ; Immunocytochemistry ; In situ mRNA hybridization ; Northern blotting ; Spermatogenic cell fractionation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Lactate dehydrogenase-X (LDH-X), a glycolytic enzyme found only in mammalian testes and spermatozoa, is encoded by a single gene (Ldh-x) in the mouse haploid genome. Several studies have demonstrated that LDH-X is associated with germ cells at specific stages of development. We have examined the expression of the Ldh-x gene during mouse spermatogenesis and testis maturation using in situ mRNA hybridization and immunocytochemistry. The results showed that transcription and translation of the Ldh-x gene are initiated at the pachytene stage of germ cell differentiation. However, although the amount of LDH-X protein increased as the germ cells progressed to maturation, its mRNA level was greatly decreased. These observations were confirmed by Northern analysis of total RNA derived from fractionated spermatogenic cells and developing testes. Furthermore, Northern studies also indicated two sizes of Ldh-x transcripts among different populations of spermatogenic cells in mature mouse testis.
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  • 9
    ISSN: 1040-452X
    Keywords: Embryonic stem cells ; Cell differentiation ; Pluripotency ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Ten embryonic stem (ES) cell lines from mink blastocysts were isolated and characterized. All the lines had a normal diploid karyotype; of the ten lines studied, five had the XX and five had the XY constitution. Testing of the pluripotency of the ES-like cells demonstrated that (1) among four lines of genotype XX, an X was late-replicating in three; both Xs were active in about one-third of cells of line MES8, and analysis of glucose-6-phosphate dehydrogenase revealed no dosage compensation for the X-linked gene; (2) when cultured in suspension, the majority of lines were capable of forming “simple” embryoid bodies (EB), and two only showed the capacity for forming “cystic” multilayer EBs. However, formation of ectoderm or foci of yolk sac hematopoiesis, a feature of mouse ES ceils, was not observed in the “cystic” EB; (3) when cultured as a monolayer without feeder, the ES cells differentiated into either vimentin-positive fibroblast-like cells or cytokeratin-positive epithelial-like cells (less frequently); neural cells appeared in two lines; (4) when injected into athymic mice, only one of the four tested lines gave rise to tumors. These were fibrosarcomas composed of fibrobalst-like cells, with an admixture of smooth muscular elements and stray islets of epithelial tissue; (5) when the ES cells of line MES1 were injected into 102 blastocyst cavities and subsequently transplanted into foster mathers, we obtained 30 offspring. Analysis of the biochemical markers and coat color did not demonstrate the presence of chimaeras among offspring. Thus the cell lines derived from mink blastocysts are true ES cells. However, their pluripotential capacities are restricted. © 1992 Wiley-Liss, Inc.
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  • 10
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 33 (1992), S. 131-140 
    ISSN: 1040-452X
    Keywords: Maturation-promoting factor ; Protein kinase activity ; Cell-free system ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Under the influence of maturation-inducing hormone (MIH) secreted from follicle cells, oocyte maturation is finally triggered by maturation-promoting factor (MPF), which consists of a homolog of the cdc2+ gene product of fission yeast (p34cdc2) and cyclin B. Two species of cyclin B clones were isolated from a cDNA library constructed from mature goldfish oocytes. Sequence comparisons revealed that these two clones are highly homologous (95%) and were found to be similar to Xenopus cyclin B1. Using monocional antibodies against Escherichia coli produced goldfish cyclin B and the PSTAIR sequence of p34cdc2, we examined the levels of cyclin B and p34cdc2 proteins during goldfish oocyte maturation induced in vitro by 17α,20β-dihydroxy-4-pregnen-3-one (17α,20β-DP), a natural MIH in fish. Protein p34cdc2 was found in immature oocyte extracts and did not remarkably change during oocyte maturation. Cyclin B was not detected in immature oocyte extracts and appeared when oocytes underwent germinal vesicle breakdown. Cyclin B that appeared during oocyte maturation was labelled with [35S]methionine, indicating its de novo synthesis. Introduction of E. coli-produced cyclin B into immature oocyte extracts induced p34cdc2 (MPF) activation. Although the possibility that immature goldfish oocytes contain an insoluble cyclin B is not completely excluded, these results strongly suggest that 17α, 20β-DP induced oocytes to synthesize cyclin B, which in turn activates preexisting p34cdc2, forming active MPF. © 1992 Wiley-Liss, Inc.
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