bluuebunny/crossref_metadata_2025 huggingface dataset:
Alterations in Lactate Dehydrogenase Isoenzymes and Phosphofructokinase Activity in the Hormonally Induced G-XII Cell Line
Authors: Malik Ajay K. , Sharma Nishi , Chandiok Preetinder , Chapekar T.N.
Published: October 2008

G-XIIcell line was earlier developed in our laboratory from the granulosa lutein cells of goat ovaries, subjected to sustained stimulation by ovine luteinizing hormone. It is a non-clonogenic and non-tumorigenic cell line which secretes progesterone. In the present paper, we report some biochemical findings on the cell line studied between passages 10 and 15. The cells exhibited a shift in the lactate dehydrogenase isoenzyme pattern from the heart form to muscle form and a 300 % increase in the phosphofructokinase activity compared to fresh granulosa cells.
Clonal Variation in the Impact of Ethyl Methanesulfonate and 5-Azacytidine on the Metastatic Phenotype of Human Prostatic Tumor Cells in Athymic Nude Mice
Authors: Ware Joy L.
Published: October 2008

Parallel in vitro exposure to 5-azacytidine or to ethyl methanesulfonate either significantly reduced or had no major impact on the experimental metastasis capacity of different clones of the human prostatic carcinoma subline 1-LN-PC-3-1A. The impact of these agents on metastatic capacity varied (1) among different clones and (2) between clones and the uncloned parental line. Analysis of the process by which these agents modify the metastatic phenotype of human tumor cells may offer insight into the process of phenotypic diversification.
Clinical Significance of Gene Amplification Studied in Human Neuroblastoma Xenografts: Relationship with Tumor Growth Rate, Chemotherapeutic Sensitivities and Levels of Neuron-Specific Enolase
Authors: Tsuchida Yoshiaki , Kanda Naotoshi , Shimatake Hiroyuki , Kaneko Yasuhiko , Notomi Tsugunori
Published: October 2008

Tumor doubling time, sensitivity to chemotherapeutic agents and concentrations of neuron-specific enolase were studied in nine human neuroblastoma xenografts, in which amplifications of N-myc, clones 8 and G21 were known; N-myc was amplified in eight, clone 8 in five and clone G21 in four of these nine xenografts. Tumor doubling time was longest in one xenograft, TNB10, which lacks the amplification of either N-myc or clone 8 or G21 and shortest in TNB1 in which all three DNA sequences are amplified with a DNA rearrangement in clone 8. No correlations were found between genomic amplification of N-myc, clones 8 and G21 and effectiveness of five chemotherapeutic drugs tested, except for cis-platinum. cis-Platinum was found to be effective on all but the one xenograft, TNB 10, with the longest tumor doubling time. Concentration of neuron-specific enolase in tumor extract was lowest in TNB1 and correlated with the length of the tumor doubling time.
Dihydroorotic Acid Dehydrogenase Activity in Actinomycin-D-Treated and Normal Chick Embryos
Authors: Katdare S.M. , Katdare Meena , Joshi P.N. , Mulherkar Leela
Published: October 2008

The effect of actinomycin D on chick embryos cultivated in vitro by New’s culturing method was studied. Exposure of chick embryos to actinomycin D (0.05 µg/ml) at the primitive streak stage (stage 4; Hamburger and Hamilton) for 6 h showed interference in orotic acid formation. The assay of the enzyme dihydroorotic acid dehydrogenase was carried out in both treated and control embryos. No enzymic activity was observed in actinomycin-D-treated embryos in contrast to the considerable activity in the controls. These observations suggest an interference by actinomycin D in the biogenesis of the enzyme dihydroorotic acid dehydrogenase.
Analysis of Cell Three-Dimensional Locomotory Vectors
Authors: Noble P.B. , Boyarsky A.
Published: October 2008

A method is presented for analysing the vectors of cells locomoting within three-dimensional collagen gels. The method detected differences in locomotory vector patterns between cells locomoting in two different gel formats. The potential of this analytical method for determining the role of the extracellular matrix in modulating cell locomotory behaviour is discussed.
Differential Distribution of B16F10 Melanoma Cells in the Liver Lobule
Authors: Hilario Enrique , Unda Fernando , Aliño Salvador F.
Published: October 2008

The distribution pattern and the number of tumor cells arrested in the liver were studied in mouse livers. Mice were perfused intravascularly with a suspension of B16F10 melanoma cells. The animals were sacrificed at 0, 1, 5 and 20 min after tumor cell perfusion. The pattern of tumor cell distribution was studied by morphological methods, and by a combined method of fluorescent-tumor cell labelling and histochemical succinate dehydrogenase activity on frozen sections, in order to define the localization of tumor cells arrested in the liver lobule. The results show that the tumor cells have an exclusive distribution in the periportal regions of the liver lobule (identified as the high succinate dehydrogenase activity areas), and that the cells are not arrested in the pericentral regions (identified as the low succinate dehydrogenase activity areas). In addition, indomethacin treatment (2 mg/kg/day) induced an increase in the number of melanoma cells arrested in the liver, but a different distribution with respect to controls was not observed. These results show that periportal regions of the liver lobule constitute a particular domain in which the B16F10 melanoma cells present a special retention ability that can be modulated by indomethacin treatment.
Transformation/Neodifferentiation of Human Skin Fibroblasts by Acute Oncornaviruses and Dexamethasone
Authors: Kopelovich Levy , Fenyk Judith
Published: October 2008

We demonstrated that the Kirsten murine sarcoma virus (KiMSV) and the Harvey murine sarcoma virus (HaMSV) converted human skin fibroblasts (HSF) into adipocytes. Adipocytic conversion of HSF by KiMSV and HaMSV was dependent on the presence of glucocorticosteroids. The Kirsten murine leukemia virus, the Harvey murine leukemia virus and the amphotropic helper virus (AP292) were ineffective by themselves. Balb murine sarcoma virus and Moloney murine sarcoma virus were, to a lesser degree, able to effect adipocytic conversion of HSF. In contrast, the feline sarcoma virus and the simian sarcoma virus did not cause this conversion. Together, the results suggest a role for certain oncogenes and glucocorticosteroids in the transformation/neodifferentiation of human cells.
Variations in WGA-Binding Proteins in Tumours Growing in Different Implantation Sites
Authors: Chan W.-S. , Page C.M. , Turner G.A.
Published: October 2008

The protein/glycoprotein composition of Triton X100 extracts of four metastatic tumour lines have been compared by gel electrophoresis after the implantation into different sites of the body. No consistent differences in the patterns were noted after staining with Coomassie Blue or treating with concanavalin A. In contrast, after treating with wheat germ agglutinin (WGA), the patterns varied with site. This was most marked with the ML and M5 tumours, where the overall binding was much lower for some sites. Although WGA binding with the LL and BL6 tumours was generally higher than with the ML and M5, reduced binding was still evident for some sites. Reduced WGA binding was also observed for extracts of secondary tumour growing in the liver or lungs.
Detection of Receptor-Mediated Membrane Effects of Dexamethasone in B16 Murine Melanoma Cell Lines by Cell Partitioning in Aqueous Two-Polymer Systems
Authors: Risely G.P. , Sherbet G.V.
Published: October 2008

Four metastatic variant sublines of the B16 murine melanoma were assayed for glucocorticoid receptors and examined for effects of dexamethasone on surface charge-related partition behaviour in aqueous two-polymer systems, expression of membrane external proteins, and adhesion to growth substratum. BL6 and F10 cells possessed cytosolic glucocorticoid receptors and, on exposure to dexamethasone, showed increased partition in the charged aqueous two-polymer system with high phosphate, but not in non-charged PO4/NaCl buffer system. This suggests that the charged two-polymer system may detect membrane changes that may be receptor-mediated effects of dexamethasone. An increase in expression of certain proteins (p250) was detected in glucocorticoid receptor-positive BL6 and F10 cells but not in the receptor-negative lines. However, other proteins, such as p220, showed an increase in all four cell lines, presumably not receptor-mediated. Dexamethasone produced no detectable changes in the ability of the cells to adhere to plastic substratum.
Cytotoxicity of N6-Cycloalkylated Adenine and Adenosine Analogs to Mouse Hepatoma Cells
Authors: Thedford Roosevelt , Leyimu Eniola O. , Thornton Dedra L. , Mehta Rajen
Published: October 2008

Cytotoxic effect(s) of N6-cycloalkylated adenine and adenosine derivatives, upon the viability of mouse hepatoma cells, were studied in vitro. N6-Cyclopropyl- and N6-cyclobutyladenine and adenosine derivatives (33 µg/ml; 24–48 h) exerted significant cytotoxic effects upon the cells. N6-Cyclopentyl- and N6-cyclohexyladenines exerted similar effects under different experimental conditions (133–166 µg/ml; 48–72 h), while no significant cytotoxic effect(s) were observed with the corresponding adenosine derivatives under these conditions. Observed physical changes in the treated cells included cell elongation, short stubby filaments, wide intracellular spaces and ruptured cell membranes. N6-Cycloal-kylated nucleosides were usually more cytotoxic than the cycloalkylated bases.
Cohesive Properties of Terminally Differentiated Keratinocytes
Authors: Brysk Miriam M. , Rajaraman Srinivasan , Penn Philip , Barlow Elisia , Bell Trace
Published: October 2008

The stratum corneum can be dissociated into single squames by several extraction methods: mechanically, with organic solvents, and with detergents. We have performed studies of reaggregation from single squames prepared by these methods. Mechanically dispersed corneocytes recombined into a lamellar-like structure closely resembling intact stratum corneum. Squames obtained by dissociation in ether formed a multilayered structure which differed from the intact tissue in the collapse of the bilayered lamellar structure into a single lucent band with osmiophilic deposits. Squames obtained after extraction with detergent failed to reaggregate.
Differences in Intracellular Calcium Mobilization by Interferon-β and Interferon-γ in RPMI-4788 Cells
Authors: Miyake Mikio , Fuchimoto Sadanori , Orita Kunzo
Published: October 2008

Human interferon (IFN) stimulates a 1.5- to 1.7-fold transient increase in the concentration of cytoplasmic-free calcium ion ([Ca2+]i) within 10–20 s upon exposure of RPMI-4788 cells to IFN. This early event of IFN-induced [Ca2+]i mobilization was measurable by loading the cells with Fura-2AM, a fluorescent Ca2+ indicator. The mobilization induced by IFN-β or IFN-γ was dependent on the concentration of each IFN. The increased [Ca2+]i gradually returned to its resting level within 60 s. The addition of EGTA (0.5–10 mM) to medium induced a marked decrease in the amount of [Ca2+]i mobilized by IFN-β and a partial decrease by IFN-γ. This finding suggests that the mechanisms of [Ca2+]i mobilization by IFN-β and IFN-γ might be different. While IFN-β-induced mobilization may be mainly from an influx of the extracellular calcium ion QCa2+]₀), IFN-γ-induced mobilization may be a summation of an influx of [Ca2+]₀ and a release from intracellular Ca2+ stores.
Behavior of in vitro Grown Normal Human Mucosal Epithelial Cells and Tumorigenic Rat Cells Inoculated into Nude Mice
Authors: Holmstrup Palle , Arenholt-Bindslev Dorthe , Jepsen Arne , Stoltze Kaj
Published: October 2008

The present study describes the behavior of in vitro grown normal human oral mucosal epithelial cells and that of a tumorigenic epithelial cell line following subcutaneous inoculation into nude mice. A successful recovery of viable human epithelial cell inocula was seen in 25–90% of mice and there was no improvement in recovery rates after addition of fibroblasts. These inocula resulted in cyst formation lined by a 2–6 cell layer unkeratinized squamous epithelium without rete ridges. There was no increase in recovery rate or size of cysts when coinoculated with fibroblasts. The tumorigenic cell inocula were successfully recovered in all cases. Tumors established from these inocula had a low grade of differentiation and were without signs of metastasis. Inocula of tumorigenic cells showed an increased size after addition of fibroblasts to the inocula. The model may be useful in studies of interactions between inoculations of heterologous normal and pathologic cells as well as in studies of differentiation of carcinogen-treated epithelial cells.
Expression of the Plasma Membrane Proteolipid in Mouse Neuroblastoma Cells: Transient Increase in Synthesis during Differentiation with N6, O2-Dibutyryl Adenosine 3’, 5’-Cyclic Monophosphate
Authors: Shea Thomas B. , Fischer Itzhak
Published: October 2008

We have examined the regulation of plasma membrane proteolipid (PM-PLP) synthesis and steady-state levels in mouse NB2a/dl neuroblastoma cells during differentiation with dibutyryl cyclic AMP (dbcAMP) and retinoic acid (RA), agents which have been previously shown to induce the elaboration of exclusively axonal or dendritic neurites, respectively. We report that a PM-PLP-immunoreactive species is expressed by this neuroblastoma cell line, and that its expression is regulated by specific states of differentiation. Differentiation of cells with dbcAMP was accompanied by an initial 2-fold increase in this PM-PLP immunoreactive species at 24 h after treatment, which returned to control levels by 96 h after treatment. By contrast, no significant increase in synthesis was detected when cells were treated with RA. Protein blot analysis of PM-PLP in dbcAMP-treated cells indicated that there was little change in its steady-state level until 96 h following treatment, at which time a reduction of 40% was observed. Throughout induced differentiation with dbcAMP, NB2a/dl cells continued to express a PM-PLP-immunoreactive species which comigrated on immunoblot analysis with PM-PLP form characteristic of embryonic brain (14–16 kDa), and apparently did not express the PM-PLP form characteristic of adult brain (18 kDa).
Mitogenic Effect of Estradiol on MCF-7 Human Breast Cancer Cells Can Be Modulated by Serum
Authors: Najid A. , Nicolas A. , Tixier M. , Habrioux G.
Published: October 2008

The MCF-7 human breast cancer cell line responds to estradiol stimulation in vitro by increased proliferation only if prolonged subcultures in dextran-coated charcoal-treated fetal calf serum have been made previously. This growth stimulation is not obtained when cells are grown in medium containing 5 % untreated fetal calf serum. We describe here the culture conditions under which we obtain a reproducible estradiol effect on cell growth.
The Role of Laminin in Cancer Invasion and Metastasis
Authors: Hunt Gillian
Published: October 2008

Laminin, a basement membrane glycoprotein, has been implicated in a number of stages in tumour invasion and metastasis. In addition to its roles in cell adhesion and migration, laminin may be important in mediating interactions of tumour cells with the immune system and have more subtle roles in controlling metastatic behaviour. Recent work on the prognostic significance of laminin and the possible role of the molecule in anti-metastasis therapy will also be discussed.
Immunoglobulin and T Cell Receptor Gene Rearrangements in Lymphoproliferative Disorders
Authors: Daus H. , Schwarze G. , Pees H. , Radtke H. , Kümel G. , Scheurlen PG.
Published: October 2008

Thirty-one samples representing Hodgkin’s and non-Hodgkin’s lymphomas, angioimmunoblastic lymphadenopathy (AILD), and benign follicular hyperplasia in HIV infections were examined for rearrangements of the immunoglobulin (Ig) and T cell receptor (TcR) β-chain gene loci. In 11 of 12 non-Hodgkin’s lymphomas (classified as Burkitt lymphoma (2), centrocytic lymphoma (1), centrocytic-centroblastic lymphoma (5), centroblastic lymphoma (3)), only rearranged Ig genes could be detected. The exceptional case was an unclassified high-grade lymphoma, which represented a rearrangement of the TcR β-chain. We also examined DNA from lymphoid neoplasms in which the lineage of the malignant cell was still controversial. Rearrangement of the TcR could exclusively be demonstrated in all 3 cases of AILD. One Ig gene rearrangement and 4 TcR β-chain rearrangements were found in 13 samples of Hodgkin’s lymphomas (11 lymph nodes, 1 pleura effusion and 1 bone biopsy with proven infiltration). Examination of 3 cases of benign follicular hyperplasia in HIV infection represented one Ig rearrangement.
Characterization and Differential Expression of an Endothelial Cell-Specific Surface Antigen in Continuous and Sinusoidal Endothelia, in Skin Vascular Lesions and in vitro
Authors: Goerdt Sergij , Steckel Friedhelm , Schuize-Osthoff Klaus , Hagemeier Hans-Hermann , Macher Egon , Sorg Clemens
Published: October 2008

Continuous and sinusoidal endothelial cells display marked morphological and functional heterogeneity as to their plasmalemmal vesicle content, to the kind of intercellular junctional complexes, to the existence and kind of fenestrae and gaps, to the existence and character of their basement membrane, to their ability for phagocytosis and to other functional parameters. Monoclonal antibody 1F10, raised against human umbilical vein endothelial cells (HUVE cells), reflects these differences in recognizing – without any nonendothelial side reactions – an endothelial cell surface antigen, abundantly expressed in continuous endothelia, low and inconsistently expressed in liver sinusoidal and dermal lymphatic endothelia and absent from splenic sinusoidal endothelial cells. In differentiated skin vascular tumors, 1F10 antigen is expressed in normal amounts while it is only low and inconsistently expressed in the dedifferentiated endothelial cells of Kaposi’s sarcoma and hemangiosarcoma. HUVE cells in culture, in contrast to their in situ ancestors, express variable amounts of 1F10 antigen. When endothelial cell-conditioned medium (ECC medium) is supplied to HUVE cells in culture, no 1F10 antigen is expressed, while supplementation with fresh serum-containing medium (FSC medium) or cytokines, such as bFGF, suffices to maintain 1F10 expression in 10–70% of the cells. From this we conclude that developmental regulation, environmental influences and cytokine supply contribute to the differentiation and maintenance of the 1F10+ and 1F10- endothelial cell phenotypes, both in vivo and in vitro.
Vascularization of Ovaries from Golden Hamsters Following Implantation into the Chick Chorioallantoic Membrane
Authors: Spanel-Borowski Katharina
Published: October 2008

In order to study the angiogenic activity, entire ovaries of immature or adult golden hamsters in different functional stages were implanted into the chick chorioallantoic membrane (CAM). Ovaries or other organs (heart, lung, liver, spleen, kidney, adrenal gland, testis) were applied during days 9–14 of incubation. Histological serial sections and ultra-structural studies displayed vascularization from the host to the ovary. Vascularization was marked for unstimulated immature ovaries compared to superstimulated immature ovaries or adult ovaries obtained during the period of proestrus-estrus. When no vascularization developed, grafted ovaries of immature or adult animals induced a mild erosive reaction of the CAM. In contrast, few other implanted organs obtained from immature animals caused a response with vascularization which was absent in grafted organs from adult hamsters. The erosive reaction of the CAM was rare, but the ulcerative response accompanied by infiltrating eosinophils was frequently seen both in immature and adult animals. These results may indicate that angiogenetic activity of the ovary is not the sole cause for vascularization of grafts. Other factors such as tissue death or a low histocompatibility barrier are likely to be involved.
Fibronectin Content of the Annulus fibrosus in Diabetic and Non-Diabetic Sand Rats
Authors: Silberberg R. , Meier-Ruge W. , Odermati B.
Published: October 2008

Annulus fibrosus of intervertebral discs from diabetic and non-diabetic sand rats were examined by microspectrophotometry for fibronectin content. This was higher in the diabetic animals both in the dorsal and ventral parts and in the outer and inner lamellae of the annulus. It is suggested that diabetes-related changes in fibronectin are similar to changes in annular collagen observed in species other than sand rats.
Time-Based Changes in Fibroblast Three-Dimensional Locomotory Characteristics and Phenotypes
Authors: Noble P.B. , Shields E.D.
Published: October 2008

Three-dimensional loçomotory trajectories have been determined for an embryonic fibroblast population and also for these cells after a 20-day period in tissue culture (C-20 cells). Differences in loçomotory characteristics between these two cell populations are reported. Applying factor analysis to the distribution of angle changes between vectors in a cell’s trajectory reveals the possibility that different loçomotory phenotypes exist. Of the three phenotypes detected in the embryonic population only one continues in the C-20 population while a new phenotype appears in the latter. These results document changes in loçomotory characteristics and phenotypes in an embryonic population with time.
Changes in Collagen Synthesis hy Human Bone Marrow Fibroblasts with Progressive Subcultivation
Authors: Fernandez Mireya , Barahona Cristina , Martinez Jorge , Minguell Jose J.
Published: October 2008

Bone marrow fibroblasts from normal and leukemic patients were used to investigate the relationship between serial subcultivation and changes in collagen synthesis. A regime was established to generate subcultures up to 35 cumulative population doublings (CPDs) in normal cells and to 9 CPDs in leukemic cells. In both types of cells, collagen synthesis decreased as subcultivation progressed. In normal cells, collagen synthesis was reduced to 10% of the original levels at 18 CPDs and in leukemic cells at 8 CPDs. In normal fibroblasts, collagen synthesis was more profoundly affected than overall protein synthesis by subcultivation. In acute lymphoblastic leukemia-derived fibroblasts, the decrease in collagen synthesis paralleled that of total protein.
Attempted Modulation of Disulfide Antitumor Activity in Balb/c Mice through Glutathione Depletion
Authors: Coshan-Gauthier R.K. , Kirkpatrick D.L.
Published: October 2008

We investigated the effects of buthionine sulfoximine (BSO)-mediated glutathione (GSH) depletion on the antitumor activity in Balb/c mice produced by four disulfide derivatives of 6-TG and 6-MP. Initial studies indicated that 14 h after BSO (5 mmol/kg) injections, tumor GSH levels were maximally depleted, while normal tissue GSH levels had returned to near control levels. Tumor growth delays and growth rates were compared for groups of animals receiving disulfides I–IV with and without BSO administration 14 h previous. Treatments with BSO alone produced no delay or growth rate differences from the control. Compounds II or III administered in the presence and absence of BSO also produced no delay or growth rate differences from control. Compound I (10 mg/kg) alone showed a delay of 5.2 days and a growth rate significantly slower than that of control (p = 0.05). In combination with BSO the effects were not enhanced. Compound IV (50 mg/kg) also produced delays in 2 separate trials (3.1 and 4.8 days) and significantly slower growth rates on each occasion compared to the control (p = 0.05). The growth rates were not significantly lowered in the presence of BSO. Administration of two doses of IV, 4 days apart, produced a delay (4.9 days) similar to that seen with a single dose. It produced 2 cures and was also more toxic, causing 3 deaths. Two doses of IV in combination with BSO pretreatment had a greater delay (16.0 days) and a significantly longer growth rate (p = 0.05) than two doses of IV alone. This treatment produced 1 cure, but also increased the number of deaths to 5/8. 6-TG (50 mg/kg) produced a delay of 11.8 days which was not enhanced with BSO. If compared to that of IV, on an equimolar ratio the delay produced by 6-TG was equivalent. BSO pretreatment, in general, was more toxic than drug alone and provided enhancement of antitumor activity only with the double dose of compound IV (50 mg/kg).
Review of the Significance of Pulmonary Intravascular Macrophages with Respect to Animal Species and Age
Authors: Winkler Gian C.
Published: October 2008

Pulmonary intravascular macrophages (PIMs) have been recognized as the site of substantial uptake of blood-borne particles in the lungs of a number of domestic animal species. Concomitantly, there is a pronounced lung susceptibility to endotoxin in calves, goats, sheep, pigs, and cats. Hemodynamic changes and initial lung injury after endotoxin administration are mediated by arachidonic acid metabolites from a pulmonary source. A significant role of PIMs in regulating pulmonary hemodynamics is implicated.
Formation of Double Contour as a Repair Reaction in the Human Femoral Head
Authors: Lothe Kari , Spycher Max A.
Published: October 2008

Thirty-two femoral heads from autopsy material with arthrotic changes and 12 surgical specimens of femoral heads with different diseases have been examined with respect to their ability to form a ‘double contour’. An additional bony plate within the cartilage, giving rise to the double contour, occurred only in the non-pressure zone of the articular cartilage. There were no major differences between autopsy and surgical material and these changes could also not be found in normal femoral heads. The origin of a double contour could be traced to tissue elements penetrating the subchondral bone from the bone marrow, and was the result of a gradual transformation with chondroid tissue as the starting material. We observed a similar transformation leading to lamellar bone in association with pannus.
Hypoxic Injury of Rat Cortical Neurons in Primary Cell Cultures
Authors: Kinoshita A. , Yamada K. , Hayakawa T.
Published: October 2008

A primary culture of cortical neurons was established from a 17-day-old rat embryo. Twenty-four hours after plating (day 0), the medium was changed to a chemically defined one. Hypoxia was exerted for 4 h by placing cultures in an air-tight chamber connected to a vacuum pump and a cylinder filled with 95 % N2 and 5 % CO2. Hypoxic stress was exerted on one of the consecutive days from day 0 to day 3. Hypoxia on days 0, 1 or 2 reduced the number of surviving neurons immediately following exposure. However, hypoxia on day 3 resulted in delayed reduction. Anitsynaptophysin antibody was used immunocytochemically to examine whether the difference is related to the synaptic formation of cultured neurons. Neurons which had more and longer neurites were clearly stained for synaptophysin. We suspect that neurons with synaptic formation release substances from their presynaptic vesicles, and that these substances might account for the delay reduction.
Analysis of Expression of Human Mammary Epithelial Antigens in Normal and Malignant Breast Cells at the Single Cell Level by Flow Cytofluorimetry
Authors: Peterson Jerry A. , Bartholomew James C. , Stampfer Martha , Ceriani Roberto L.
Published: October 2008

The expression of cell-type specific antigens on the surface of human mammary epithelial cells (HME antigens) is analyzed at the single cell level by flow cytofluorimetry. Methods are described to deal with two technical difficulties that have hampered the quantitation of surface antigens on cells from primary cultures by flow cytofluorimetry, namely, cell clumping and the presence of several cell types. By staining the cells simultaneously with two different fluorescent markers: the cell surface HME antigens with fluorescein-tagged antibodies (green fluorescence) and the cell DNA with propidium iodide (red fluorescence), it was possible to determine the expression of HME antigens per unit DNA, analyze their expression on breast cells in mixed populations, and analyze their expression in different phases of the cell cycle. The HME antigens were diminished in malignant breast epithelial cells as compared to their normal counterpart.
Inhibition of Mitogen and Specific Antigen-Induced Human Lymphocyte Proliferation by Cadmium
Authors: Kaštelan Maja , Gerenčer Marijan , Kaštelan Andrija , Gamulin Stjepan
Published: October 2008

Cadmium chloride (CdCl2) added to human lymphocyte culture inhibits the proliferative response induced by phytohaemagglutinin (PHA), pokeweed mitogen and allogenic lymphocytes in mixed lymphocyte reaction. Minimally effective concentrations of CdCl2 were 3.3, 1.6 and 1.6 µM, respectively. The inhibition was greatest when CdCl2 was added at initiation of cultures and declined if the addition of CdCl2 was postponed. The presence of CdCl2, regardless of the presence of PHA during the first 24 h of incubation suppressed the proliferative response to subsequent stimulation with PHA, indicating that cadmium affects an early step of blastogenic transformation.
Synovial Cell Antigens – Production of Heterologous Anti-Human Synovial Cell Sera and General Reactivities of the Antisera
Authors: Sloan Tod B. , Martincic Rasa R. , Anderson Byron
Published: October 2008

Antisera to cultured human synovial cells were produced in several rabbits by a variety of immunization procedures. The antisera were assayed by a number of immunologic techniques for specificities against live synovial and other fibroblastic cell lines and against additional antigen sources including human serum and synovial fluid, and fetal calf serum. While the antisera reacted well and with the same titers to all synovial cell lines, reactivities varied with several nonsynovial cell lines showing that synovial tissue-derived cells are antigenically different from other tissue fibroblastic cell strains. Also, the major antigens cross-reactive between synovial and certain other fibroblastic cell lines were partially defined as those in or cross-reactive with fetal calf serum and human serum or plasma components. The differential reactivities of the antisera demonstrated that the antibody specificity populations were markedly affected by the immunization manipulations. Methods for analyzing those differences are presented by defining specificity and reactivity indices. The antisera to the synovial cells distinguished 7 electrophoretically distinct reactivities in human serum or plasma, synovial fluid, and in fetal calf serum. 1 antigen was identified as fibronectin, 3 as components of fetal calf serum and 3 additional unidentified human serum cross-reactive antigens.
Study of Agglutination of Mouse Mammary Carcinoma (FM3A) Cell Induced by Egg Agglutinin of Rana catesbiana
Authors: Takeda S. , Kubota K. , Endo Y. , Matsuzawa T.
Published: October 2008

When mouse mammary carcinoma (FM3A) cells were treated with egg agglutinin of Rana catesbiana for 15 min at 25 °C, percent total particle number of both cell aggregates and single cells was in direct proportion to the cell electrophoretic mobility. Phytohemagglutinin P mediated agglutination proceeded with biphasic kinetics: the higher the concentration of phytohemagglutinin Pthe shorter was the lag period between the first and second stages of agglutination. In protamine-mediated agglutination, the percent total particle number was reduced at low concentrations, while the electrophoretic mobility reduced only at high concentrations. Agglutinating and cytotoxic activities of these three reagents were in an intimate relation: the higher the agglutinating activity, the greater was their cytotoxic activity.
Alpha-Fetoprotein and Albumin Synthesis by Heterotransplanted Rat Liver Tumor Cells
Authors: Higgins Paul J. , Borenfreund Ellen
Published: October 2008

Epithelial cells, isolated from the hepatic tissue of rats bearing carcinogen-induced neoplastic liver nodules, were analyzed by immunological methods for the production of specific liver proteins during alternate in vitro-in vivo passage. Heterotransplantation of several long-term rat liver cell lines into nude mice and subsequent re-establishment in vitro resulted in the derivation of hepatocyte cultures expressing both alpha-fetoprotein and albumin. Prior to passage in vivo these cell lines synthesized only one or neither of these proteins in vitro. Alternate in vitro-in vivo passage thus appears to potentiate tissue-specific protein expression by established liver cell lines.
Interactions between Human Lymphoblastoid Cells and Human Fibroblast Feeder Layers in vitro
Authors: Delinassios John G. , Kottaridis Stavros D.
Published: October 2008

Lymphoblastoid (LB) cells interact in vitro with human fibroblasts. Cell complexes can be dissociated by trypsin treatment. Binding on fibroblasts is higher for Epstein-Barr virus-producer LB cells than for nonproducers, and depends on the origin of the fibroblastic cells. Stromal fibroblasts isolated from a metastatic lymph node tumor exhibited a highly increased potency of binding LB cells in comparison with fibroblasts isolated from normal breast epidermis or the primary breast tumor from the same patient. After a 10-hour interaction period, LB cells undergo abnormal divisions giving rise to the formation of ‘mini cells’.
Glycoprotein Enrichment in Moloney Leukemia Virus Structural Proteins Released from Interferon-Treated Cells
Authors: Sherman Levana , Teitz Yael
Published: October 2008

Interferon treatment of Moloney-leukemia-virus-infected cells (3T3/MLV) leads to the formation of virus particles enriched with viral structural glycoproteins, in addition to the inhibition of virus production. A preferential inhibitory effect on incorporation of RNA and proteins rather than glycoproteins was found in the released virus particles from interferon-treated cells. Enrichment in 70,000- and 45,000-dalton glycoprotein (gP-70, gP-45) in these particles was further demonstrated by polyacrylamide analysis of viral proteins pulse-labeled with [3H]-leucine. Viral glycoproteins released as soluble antigens were also determined. A 40% reduction was found in gP-70 and gP-45 released from interferon-treated cells. Radioimmunoprecipitation of pulse-chase-labeled cellular viral proteins showed no effect of interferon on the formation of viral structural 30,000-, 15,000- to 12,000-dalton proteins, and gP-70 and gP-45 from their respective precursors. The uncoordinate effect of interferon inhibition on viral 30,000-dalton protein and gP-70 is discussed.
Synovial Cell Antigens – Differences of Antigen Compositions between Rheumatoid Arthritic (RA)- and Non-RA-Derived Synovial Cells Detected with Anti-Synovial Cell Sera
Authors: Sloan Tod B. , Martincic Rasa R. , Anderson Byron
Published: October 2008

A number of antisera were produced in rabbits immunized with cultured synovial tissue-derived cells. The synovial cells were injected utilizing a variety of immune manipulations in order to elicit different antibody specificities and to enhance antibodies that may differentiate antigens of rheumatoid arthritic and nonrheumatoid arthritic synovial tissue-derived cells. The major sources of antigens common to both rheumatoid and nonrheumatoid cells were cross-reactive with normal human serum, nonrheumatoid synovial fluids and fetal calf serum components. Adsorptions with the latter antigen sources yielded different antisera with greater differential reactivities to either rheumatoid or nonrheumatoid synovial cells. Crossed immunoelectrophoretic analyses of extracts of the synovial cells using unabsorbed antisera differentiated five electrophoretically distinct antigens with various quantitative differences of the antigens being noted between the rheumatoid and nonrheumatoid extracts. The results suggest that several antigens are quantitatively differentially expressed on rheumatoid-derived synovial cell strains compared to synovial strains of nonrheumatoid origin.
Distribution of Nuclear Size and DNA Content in Serial Liver Biopsies of Rats Treated with N-Nitrosomorpholine, Phenobarbital and Butylated Hydroxytoluene
Authors: Romagna F. , Zbinden G.
Published: October 2008

Nuclear size distribution and DNA content distribution were determined in liver biopsies of rats using an electronic particle counter and an impulse cytophotometer, respectively. Nuclear size distribution and DNA content distribution were found to be highly correlated regardless whether peak areas or peak heights of the histograms were compared. Treatment with the hepatocarcinogen NNM caused a dose-dependent increase in octoploid nuclei and a shift of the 2 n : 4 n ratio in favor of the diploid nuclei. Treatment with Pb and BHT had minor effects on nuclear size and DNA content distribution. The paper demonstrates the usefulness of the fine-needle aspiration biopsy procedure for the evaluation of liver changes induced by chemical substances.
Association between HLA-A Alleles and Alzheimer’s Disease in a Southern Chinese Community
Authors: Ma Suk Ling , Tang Nelson Leung Sang , Tam Cindy Woon Chi , Lui Victor Wing Cheong , Suen Eddie Wai Chui , Chiu Helen Fung Kum , Lam Linda Chiu Wa
Published: October 2008

Background/Aim: HLA-A is a locus of the major histocompatibility complex situated on chromosome 6p21.3. Several genome-wide linkage analyses suggested that a putative locus predisposition for Alzheimer’s disease (AD) is found on chromosome 6p21. In this study, we investigated the association between HLA-A alleles and AD in a Chinese community. Methods: Fine genotyping of 160 Chinese AD and 167 age-matched nondemented subjects was performed for the first time by sequence-based typing of the HLA-A locus. Results: We found that HLA-A2 carriers (including prevalent alleles, 0201, 0203 and 0207) were more prevalent in the AD group; the frequency of HLA-A2 was increased in AD patients versus controls, but the difference was not significant after Bonferroni correction for the number of alleles tested (p = 0.075). A gene-gene interaction was observed between ApoE and HLA-A, the presence of HLA-A24 (particularly the prevalent allele 2402) might act as another independent risk factor of developing AD for subjects not carrying the ApoE Ε4 allele (relative risk = 2.98, 95% CI = 1.14–8.24). Carriers of HLA-A2 had an earlier onset of AD by 2.4 years (p = 0.030) and HLA-A2 interacted with ApoE Ε4 in modulating the age at onset in AD. Conclusions: This study suggested that HLA-A may be involved in the pathogenesis of AD.
Dialysis and the Elderly: An Underestimated Problem
Authors: Buemi Michele , Lacquaniti Antonio , Bolignano Davide , Donato Valentina , Fazio Maria Rosaria , Campo Susanna , Coppolino Giuseppe , Sturiale Alessio
Published: October 2008

In developed countries, the incidence of end-stage renal failure is constantly increasing, and uremia will soon be a disease typically found in mature and elderly adults. Almost invariably, the physical condition of the elderly patient with terminal uremia is extremely poor, and therapeutic approach complex. Frequent co-morbidity, treatment with many different drugs, the high risk of iatrogenic damage, advanced age and socio-environmental conditions further complicate the management of these patients. While replacement therapy may become necessary, peritoneal dialysis may have advantages over hemodialysis. Peritoneal dialysis causes less hemodynamic stress, does not necessitate vascular access and allows mobility, although it incurs a high incidence of peritonitis and vascular disease. Where hemodialysis is the only feasible treatment, procedures used for vascular access are frequently followed by several complications, representing an important cause of morbidity and hospitalization. In addition, even if it may improve the patient’s quality of life, vascular condition, intradialytic hypotension, heart disease, intestinal bleeding and amyloidotic arthropathy are critical aspects of dialysis in the elderly patient. Therefore, particular attention from clinicians and administrators is required and the best possible strategies must be identified in order to provide effective and appropriate services to address these special patients’ needs.
Plasma Antioxidants and Asymptomatic Carotid Atherosclerotic Disease
Authors: Riccioni Graziano , Bucciarelli Tonino , D’Orazio Nicolantonio , Palumbo Nicola , di Ilio Emanuela , Corradi Francesco , Pennelli Alfonso , Bazzano Lydia A.
Published: October 2008

Background: Atherosclerosis remains clinically mute for a long time and frequently manifests itself with an acute cardiovascular event. The possibility of detecting this disease in a subclinical phase and reducing or reversing its progression is an issue of relevance. Published studies on the association between antioxidant vitamins and carotenoids and carotid intima-media thickness (CIMT) have been inconclusive. Methods: We enrolled 220 consecutive, asymptomatic participants. After carotid ultrasound investigation, a medical history was taken, a physical examination was performed and venous blood samples were collected. Venous blood samples were analyzed for concentrations of antioxidant vitamins and carotenoids. Results: Low concentrations of vitamin A (p < 0.01), vitamin E (p < 0.001), lycopene (p < 0.01) and β-carotene (p < 0.001) were significantly associated with carotid atherosclerosis (CIMT ≧0.8 mm). In addition, marginally higher body mass index, plasma haemoglobin and high-density lipoprotein cholesterol were also associated with carotid atherosclerosis, while other laboratory parameters considered in this study (total cholesterol, low-density lipoprotein cholesterol, triglycerides and C-reactive protein) were not significantly associated with carotid atherosclerosis. Conclusions: Low plasma concentrations of antioxidant vitamins (A, E, β-carotene) and lycopene were associated with early carotid atherosclerotic lesions as measured by CIMT. Regular intake of foods rich in lycopene and antioxidant vitamins may slow the progression of atherosclerosis.
A Golgi Silver Study on the Development of Pyramidal and Stellate Cells of the Human Cortex
Authors: Tiu S.C. , Yew D.T.
Published: October 2008

Silver impregnation studies of neurons in the developing human nervous system had attracted only a fair amount of studies due to the difficulty of the technique. This report is concerned with studying the morphogenesis of pyramidal and stellate cells during development in different regions of the cortex. No silver-impregnated cells were present before 21 weeks of gestation in most cortices except for the frontal cortex. Between 21 and 25 weeks, pyramidal cells were clearly demonstrated. By 27 weeks of gestation and thereafter, secondary branching was observed in these cells. Two types of stellate cells were identified at week 30 and thereafter. A difference in the degree of differentiation was also recorded between different cortical regions during development.
Antithrombotic Effects due to Pharmacological Modulation of Thrombin-Activatable Fibrinolysis Inhibitor in Rats
Authors: Soni Hitesh , Sharma Ajay , Bhatt Shvetank , Jain Mukul R. , Patel Pankaj R.
Published: October 2008

Thrombin-activatable fibrinolysis inhibitor (TAFI) is a basic carboxypeptidase zymogen that can be activated by thrombin. Activated TAFI (TAFIa) cleaves carboxyl-terminal lysine residues from partially degraded fibrin, rendering it resistant to fibrinolysis by endogenous tissue plasminogen activator (tPA). Carboxypeptidase inhibitor (CPI) isolated from potato inhibits TAFIa and reduces clot lysis time in rabbit and mouse plasma. In the present study, we report the effect of CPI on tPA-mediated clot lysis using rat plasma in vitro. CPI at 400, 600 and 800 ng/ml caused a dose-dependent enhancement of tPA-induced clot lysis. In vivo effect of CPI was also investigated using ferric chloride-induced arterial thrombosis model in rat. The results showed that i.v. administration of CPI significantly prolonged the ‘time to occlusion’ at the dose of 2 and 4 mg/kg. At 2 mg/kg i.v. dose in rat, CPI showed no effect on prothrombin time and activated partial thromboplastin time, indicating noninterference of CPI with other clotting factors in mediating its thrombolytic effect through TAFI inhibition. Furthermore, 2 mg/kg i.v. dose of CPI did not produce significant increase in bleeding time when tested in rat tail-transection bleeding model. These results provide evidence for a role of TAFI in arterial thrombosis in rats and suggest that TAFI inhibition could be explored as an attractive target for the development of new antithrombotic drugs.
Sleep Apnea in Morbidly Obese Patients: Prevalence and Clinical Predictivity
Authors: Palla Antonio , Digiorgio Michela , Carpenè Nicoletta , Rossi Giuseppe , D’Amico Ilaria , Santini Ferruccio , Pinchera Aldo
Published: October 2008

Background: Little is known about the prevalence of obstructive sleep apnea-hypopnea syndrome (OSAHS) in morbidly obese patients and whether such patients show peculiar clinical findings that may make it easier to suspect and diagnose OSAHS. Objectives: To investigate prevalence of OSAHS in patients with morbid obesity and find a simple structured model for predicting the results of polysomnography. Methods: The study enrolled a group of 101 consecutive inpatients (33 males, age range 20–80 years) with a body mass index ≥40, whose symptoms of OSAHS were not known, and a validation group of 45 patients. Results: Habitual snoring, nocturnal apneas or awakening as well as diurnal sleepiness were frequent findings (90.1, 40.6, 50.5 and 61.4%, respectively). Chronic obstructive pulmonary disease, hypertension, diabetes and myocardial ischemia were also frequently associated (22.8, 56.4, 30.7 and 6.9%, respectively). OSAHS was found in 61 (60.4%) patients, in 33.7% it was of severe degree. A multivariate logistic regression model allowed to select the independent predictors of OSAHS: age, male sex, diurnal sleepiness and the value of minimum nocturnal saturation. Sensitivity of 97%, specificity of 77% as well as positive and negative predictive values of 87% and 95%, respectively, were obtained; similar results were found in the validation group. When the best obtainable cutoff on the receiver operating characteristic curve is below 40%, the instrumental diagnosis might be excluded in as many as 33% of cases, since they are not affected by OSAHS or have OSAHS of mild degree. Conclusions: OSAHS is present in almost two thirds of morbidly obese patients. By applying the prediction model we propose, one may calculate the probability of a morbidly obese patient of being affected by OSAHS.