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  • 1
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 237 (1993), S. 506-511 
    ISSN: 0003-276X
    Keywords: Anterior pituitary ; Domestic chicken ; Lipid cells ; Ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A combination of light and electron microscopy techniques was applied to characterize a luteinizing hormone (LH)-gonadotroph-like cell in the anterior pituitary gland of the adult domestic chicken. This cell type (mean ± sem, 91 ± 8 μm2) was larger than typical LH-gonadotrophs (71 ± 5 μm2, P 〈 0.01) and seen in pituitary glands from adult males but not those from adult females of the same age. The ultrastructural features of these cells were similar to typical LH-gnoadotrophs in the same section, except for the presence of large (1,126 ± 77 nm diameter), round, or polymorphic electron-dense inclusions (14 ± 3 per cell) in the cisternae of the endoplasmic reticulum. When resin sections of anterior pituitary gland were stained with the vapours from the fixative osmium tetroxide and a solution of potassium ferricyanide, inclusions were found that appeared to be lipid in nature and were circumscribed by the cisternal membranes of the endoplasmic reticulum. These lipid inclusions were associated occasionally with acid phosphatase and lysosome-like bodies. The cellular pigment, lipofuscin, considered to be a marker of aging, was absent from the anterior pituitary glands from these adult chickens. These observations suggest that the appearance of lipid in LH-gonadotrophs is unlikely to be age-related and an alternative explanation is discussed in relation to a sex difference in the LH response of adult chickens to gonadotrophin-releasing hormone (GnRH). © 1993 Wiley-Liss, Inc.
    Additional Material: 4 Ill.
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  • 2
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: This study applies terms and methods for describing spatial interactions between multivariate spatial point patterns, which are, to our knowledge, new in neurobiology. We consider two categories of points, type 1 and 2, distributed within a certain reference volume (such as a nucleus of the brainstem or a cortical area). The points may, for example, represent different categories of labelled cells or axonal fields of termination. We say that there is spatial neutrality between points of type 1 and 2 if the types are signed by random labelling. If a mechanism drives the two point categories together, we say that the point patterns are positively associated. Conversely, if a mechanism drives type 1 and 2 points apart, we say that they are segregated. By comparing two cumulative distribution functions of distances between points, we can distinguish neutrality, positive association, and segregation. One function, H12(t), is the cumulative distribution function of the distance t between a pair of randomly selected points of type 1 and 2. The other, H00(t), is the corresponding function for a pair of points randomly selected without reference to type. Plots of the estimated difference between these two functions give an indication of positive association, neutrality, or segregation. A statistical test, based on simulations of random (neutral) distributions, can be used to see whether deviations from neutrality are significant.We apply the analysis described above to a major pathway of the brain, namely the ponto-cerebellar projection. Different types of cells in the pontine nuclei are retrogradely labelled with the fluorescent tracers Rhodamine-B-isothiocyanate, Fluoro-Gold, and Fast Blue. The tracers are injected in adjacent or more distant folia of the cerebellar paraflocculus. The location of the somata of labelled cells are recorded and the total distribution reconstructed in three dimensions and displayed on a dynamic graphics workstation. We ask whether different units (folia) in the paraflocculus receive information from the same population, from two different positively associated populations, or from segregated cell populations. We find a statistically significant tendency for cell populations projecting to adjacent folia to be positively associated, although there are few cells containing multiple labels. Populations of neurons projecting to folia wider apart are significantly segregated. From inspections of the reconstructions, using real-time rotations, we find that the swarms of labelled neurons tend to accumulate in shells or lamellae in the pons. Within the lamellae, the cells are aggregated in clusters and bands with empty holes (containing unlabelled ponto-cerebellar cell bodies, presumably projecting to other cerebellar targets) in between. By determining the average distance to a reference plane for each cell population, we find that cell populations shift in a ventro-medial direction as the injection sites move from the medial part of the dorsal paraflocculus toward the lateral part and into the ventral paraflocculus. We therefore conclude that there is a continuous shift in location of ponto-cerebellar cell populations, corresponding to specific shifts in cerebellar target regions.
    Additional Material: 10 Ill.
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  • 3
    Electronic Resource
    Electronic Resource
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 50 (1992), S. 69-73 
    ISSN: 0730-2312
    Keywords: DRE ; predicted cancer volume ; prostate cancer ; PSA ; TRUS ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: In a study population, can digital rectal examination (DRE), transrectal ultrasound (TRUS), and prostate specific antigen (PSA) (monoclonal) effectively detect the majority of clinically relevant cancer? If this is possible, the remaining patients could then be considered for chemopreventive protocols.The American Cancer Society/National Prostate Cancer Detection Project (ACS/NPCDP) had a cancer detection rate of 2.4% for its initial year utilizing PSA, DRE and TRUS. TRUS and PSA detected 73% more cancer than DRE alone. TRUS detected a greater percentage of cancers than DRE (85% vs. 64%).PSA was ≥ 4 ng/ml for 66% of prostate cancer patients; 11% of cancer patients had PSA 〈 2 ng/ml. PSA decision levels based on gland volume detected a subgroup at the 95th percentile that had a nine-fold increased risk for cancer. In a separate study differentiating benign prostatic hypertrophy (BPH) and cancer, we found 0.12±0.13 ng/ml/gm for serum PSA (sPSA)/gm BPH. This study proved that predicted PSA (pPSA) = gland volume × 0.12; this equation also functioned at the 95th percentile for any individual patient.Individual patient assessment: 1Entry level PSA = 2 ng/ml.2Those patients with PSA 〉 2 ng/ml have TRUS determination of gland volume (performed by technician).3pPSA = gland volume × 0.12. If sPSA 〉 pPSA then:4(sPSA-pPSA)/2 = predicted volume (cc) of cancer;53√ volume of cancer = mean diameter (cm) of cancer.Thus, these results should detect the majority of clinically relevant cancer (〉0.5cc). PSA combined with TRUS and DRE can identify high risk groups for cancer. © 1992 Wiley-Liss, Inc.
    Additional Material: 4 Tab.
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  • 4
    Electronic Resource
    Electronic Resource
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 55 (1994), S. 287-299 
    ISSN: 0730-2312
    Keywords: osteocyte ; gap junction ; cytoskeleton ; extracellular matrix ; osteocytic osteolysis ; bone membrane ; functional adaptation ; mechanical loading ; strain ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Although the structural design of cellular bone (i.e., bone containing osteocytes that are regularly spaced throughout the bone matrix) dates back to the first occurrence of bone as a tissue in evolution, and although osteocytes represent the most abundant cell type of bone, we know as yet little about the role of the osteocyte in bone metabolism. Osteocytes descend from osteoblasts. They are formed by the incorporation of osteoblasts into the bone matrix. Osteocytes remain in contact with each other and with cells on the bone surface via gap junction-coupled cell processes passing through the matrix via small channels, the canaliculi, that connect the cell body-containing lacunae with each other and with the outside world. During differentiation from osteoblast to mature osteocyte the cells lose a large part of their cell organelles. Their cell processes are packed with microfilaments. In this review we discuss the various theories on osteocyte function that have taken in consideration these special features of osteocytes. These are (1) osteocytes are actively involved in bone turnover; (2) the osteocyte network is through its large cell-matrix contact surface involved in ion exchange; and (3) osteocytes are the mechanosensory cells of bone and play a pivotal role in functional adaptation of bone. In our opinion, especially the last theory offers an exciting concept for which some biomechanical, biochemical, and cell biological evidence is already available and which fully warrants further investigations. © 1994 Wiley-Liss, Inc.
    Additional Material: 4 Ill.
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  • 5
    ISSN: 0730-2312
    Keywords: CALC gene family ; genomic organization ; transcription regulation ; biosynthesis ; islet β-cell ; insulin resistance ; islet amyloid ; type 2 diabetes mellitus ; animal model ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Islet amyloid polypeptide (“amylin”) is the major protein component of amyloid deposits in pancreatic islets of type 2 (non-insulin-dependent) diabetic patients. Islet amyloid polypeptide consists of 37 amino acids, is co-produced and co-secreted with insulin from islet β-cells, can act as a hormone in regulation of carbohydrate metabolism, and is implicated in the pathogenesis of islet amyloid formation and of type 2 diabetes mellitus. Rat islet amyloid polypeptide differs from human islet amyloid polypeptide particularly in the region of amino acids 25-28, which is important for amyloid fibril formation. In rat and mouse, diabetes-associated islet amyloid does not develop. To study the genetic organization and biosynthesis of islet amyloid polypeptide, we have isolated and analyzed the human and rat islet amyloid polypeptide gene and corresponding cDNAs. Both genes contain 3 exons, encoding precursor proteins of 89 amino acids and 93 amino acids, respectively. Apart from a putative signal sequence, these precursors contain amino- and carboxy-terminal flanking peptides in addition to the mature islet amyloid polypeptide. To understand regulation of islet amyloid polypeptide gene expression, we have identified several potential cis-acting transcriptional control elements that influence β-cell-specific islet amyloid polypeptide gene expression. Using antisera raised against synthetic human islet amyloid polypeptide we developed a specific and sensitive radioimmunoassay to measure levels of islet amyloid polypeptide in plasma and tissue extracts. Also antisera raised against the flanking peptides will be used in studying human islet amyloid polypeptide biosynthesis. Elevated plasma islet amyloid polypeptide levels have been demonstrated in some diabetic, glucose-intolerant, and obese individuals, as well as in rodent models of diabetes and obesity. To examine the potential role of islet amyloid polypeptide overproduction in the pathogenesis of islet amyloid formation and type 2 diabetes, we generated transgenic mice that overproduce either the amyloidogenic human islet amyloid polypeptide or the nonamyloidogenic rat islet amyloid polypeptide in their islet β-cells. Despite moderately to highly (up to 15-fold) elevated plasma islet amyloid polypeptide levels, no marked hyperglycemia, hyperinsulinemia or obesity was observed. This suggests that chronic overproduction of islet amyloid polypeptide “per se” does not cause insulin resistance. No islet amyloid deposits were detected in mice up to 63 weeks of age, but in every mouse producing human islet amyloid polypeptide (as in man), accumulation of islet amyloid polypeptide was observed in β-cell lysosomal bodies. This may represent an initial phase in intracellular amyloid fibril formation. The human islet amyloid polypeptide overproducing transgenic mice model offers a unique opportunity to study the biosynthesis, intracellular handling, secretion, and extracellular handling of human islet amyloid polypeptide in vivo. © 1994 Wiley-Liss, Inc.
    Additional Material: 7 Ill.
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  • 6
    Electronic Resource
    Electronic Resource
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 53 (1993), S. 135-144 
    ISSN: 0730-2312
    Keywords: urokinase plasminogen activator ; urokinase plasminogen activator receptor ; plasminogen activator inhibitor ; tissue plasminogen activator ; breast cancer ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The malignant potential of solid tumors is related to the ability to invade adjacent tissue and to metastasize. These properties of cancer cells depend on the synthesis of proteolytic enzymes which are able to digest adjacent connective tissue and basement membranes. We hypothesized that all elements of the plasminogen activation system might be overexpressed in malignant human breast tumors, functioning as an essential element in tumor invasion and metastasis. As determined by histopathological methods, the malignant tumors showed statistically significantly higher expression of urokinase plasminogen activator (uPA), type-1 plasminogen activator inhibitor (PAI-1), and especially urokinase plasminogen activator receptor (uPAR) than benign tissues. All those elements were present in higher amounts in the cancer cells than in the cells of benign or normal breast tissues. High exhibition of tissue plasminogen activator (tPA) found in cancer seems to be random and not related to the malignant or benign state, since benign and malignant tumors show overexpression of tissue plasminogen activator with similar frequency. When the tumors express high amounts of uPA, they express a high amount of uPAR in 50% of cases and PAI-1 in 57.3% of cases. When urokinase is expressed in low amount, the receptor is low in 28.6% and inhibitor in 21.4% of malignant breast tumors. This statistically significant consensus, 78.6% in the case of urokinase and its receptor and 78.6% in case of urokinase and its inhibitor, suggests that these activities may be the result of a unique mechanism of control, activated in the last steps of malignant transformation.
    Additional Material: 6 Ill.
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  • 7
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 32 (1992), S. 136-144 
    ISSN: 1040-452X
    Keywords: Growth factors ; Induction ; Embryogenesis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Establishment of the body pattern in all animals, and especially in vertebrate embryos, depends on cell interactions. During the cleavage and blastula stages in amphibians, signal(s) from the vegetal region induce the equatorial region to become mesoderm. Two types of peptide growth factors have been shown by explant culture experiments to be active in mesoderm induction. First, there are several isoforms of fibroblast growth factor (FGF), including aFGF, bFGF, and hst/kFGF. FGF induces ventral, but not the most dorsal, levels of mesodermal tissue; bFGF and its mRNA, and an FGF receptor and its mRNA, are present in the embryo. Thus, FGF probably has a role in mesoderm induction, but is unlikely to be the sole inducing agent in vivo. Second, members of the transforming growth factor-β (TGF-β) family. TGF-β2 and TGF-β3 are active in induction, but the most powerful inducing factors are the distant relatives of TGF-β named activin A and activin B, which are capable of inducing all types of mesoderm. An important question relates to the establishment of polarity during the induction of mesoderm. While all regions of the animal hemisphere of frog embryos are competent to respond to activins by mesoderm differentiation, only explants that include cells close to the equator form structures with some organization along dorsoventral and anteroposterior axes. These observations suggest that cells in the blastula animal hemisphere are already polarized to some extent, although inducers are required to make this polarity explicit.How do inducing factors affect the differentiation of the responsive tissue? One approach to this question has been to look for gene expression in response to induction, especially the activation of regulatory loci like homeobox genes. Several homeobox-containing genes including Mix.1, Xhox3, X1Hbox1, and X1Hbox6, goosecoid and members of a new class of genes named Xlim, are activated by inducing factors with different patterns of expression in the embryo. Differential expression of regulatory genes probably controls the formation of distinct tissues in an orderly pattern during embryogenesis. © 1992 Wiley-Liss, Inc.
    Additional Material: 7 Ill.
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  • 8
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 35 (1993), S. 189-196 
    ISSN: 1040-452X
    Keywords: Sperm ; Sexing ; H-Y antigen ; Anti-H-Y monoclonal antibodies ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Studies designed to answer the question whether or not H-Y antigen is preferentially expressed on Y chromosome bearing sperm have resulted in conflicting results. This is probably due to the absence of reliable methods for estimating the percentage of X and Y chromosome bearing sperm in fractions, enriched or depleted for H-Y antigen positive sperm. In recent years a reliable method for separating X and Y chromosome bearing sperm has been published. With this method, separation is achieved by using a flow cytometer/cell sorter, which detects differences in DNA content. This technique provided the first opportunity for testing anti-H-Y antibody binding to fractions enriched for X and Y chromosme bearing sperm, directly. A total of 7 anti-H-Y monoclonal antibodies were tested using sorted porcine sperm and in one experiment also sorted bovine sperm. All monoclonal antibodies bound only a fraction of the sperm (20 to 50%). However, no difference in binding to the X and Y sperm enriched fractions was found. Therefore, the present experiments do not yield evidence that H-Y antigen is preferentially expressed in Y chromosome bearing sperm. © 1993 Wiley-Liss, Inc.
    Additional Material: 3 Ill.
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  • 9
    ISSN: 1059-910X
    Keywords: Cristae ; 3D structure ; Hepatocytes ; Fibroblasts ; Adrenal cortex ; Brown fat ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Rat adrenal cortex was processed for high resolution scanning electron microscopy (HRSEM) to confirm tubular cristae, reported by transmission electron microscopy to be present in cortex mitochondria. Mitochondria in several other tissue and cell types were also observed and their ultrastructure confirmed by using three-dimensional, stereo, high resolution scanning electron microscopy. The mitochondria in rat and human hepatocytes as well as human skin fibroblasts mitochondria proved to be long, up to 46 micrometers and branching, as compared to those in liver which were spherical in shape. Cold adapted brown fat cells were packed with mitochondria, these containing plate or shelf-like cristae. Branched, rat striated muscle mitochondria were observed to curve around contractile protein filament bundles. The muscle mitochondrial cristae were found to be both tubular and plate-like, within the same mitochondrion. The ratio of tubular cristae to plate-like cristae varied considerably between muscle mitochondria. In order to use ultrastructural changes in mitochondria for differential diagnosis, and because 3D reconstruction of mitochondria based on transmission electron microscopy serial sections is severely limited in resolution, it is imperative to first develop a correct understanding of tissue specific, normal mitochondrial ultrastructure based on three-dimensional, HRSEM methods. © 1994 Wiley-Liss, Inc.
    Additional Material: 10 Ill.
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  • 10
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 26 (1993), S. 260-271 
    ISSN: 1059-910X
    Keywords: Cytoskeleton ; Microtubules ; Intermediate filaments ; Membranous organelles ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Kidneys of anesthetized rats were perfused with digitonin to extract cytosolic proteins of glomerular podocytes so that the remaining intracellular structures could be examined by three-dimensional stereo high-resolution scanning electron microscopy (HRSEM). Cytoskeleton, consisting of microtubules and intermediate filaments, was preserved with each applied concentration of digitonin. High concentrations of digitonin (1.0 mg/ml) produced a corrugated appearance in plasma membranes likely due to the formation of digitonin-cholesterol complexes. At 1.0 mg/ml digitonin, the Golgi complex became vesicularized, and mitochondria were well extracted and their ultrastructure preserved. Lower concentrations of digitonin (0.1 and 0.2 mg/ml) were less disruptive to both the plasma membrane and the Golgi complex. Mitochondria, rough endoplasmic reticulum, coated vesicles, nuclear membrane, and chromatin were well preserved. Extraction with digitonin, at the optimal concentration and perfusion time, simultaneously maintains both the cytoskeleton and membranous organelles inside the cell and provides a method to elucidate the interactions between these two components. Furthermore, digitonin extraction should preserve antigenic sites, thereby allowing the localization of intracellular proteins by backscattered electron imaging of immunogold labels in the scanning electron microscope. © 1993 Wiley-Liss, Inc.
    Additional Material: 12 Ill.
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