bluuebunny/crossref_metadata_2025 huggingface dataset:
Early Recognition of Transplantation Antigens by T Lymphocytes
Authors: Ramseier Hansruedy
Published: October 2008

In crude supernates of interactions of parental strain T spleen cells with H-2 antigen-bearing F1 hybrid spleen cells, a product (soluble early product of immune recognition, SEPIR) can be found which enhances mixed leukocyte culture (MLC) reactions set up with suboptimal cell concentrations (SMLC). Supernates of parental T cells reacting with F1 spleen cells also produce this factor. The product ameliorates SMLC proliferative responses specifically since identical antigenic specificities have to be recognized in both SEPIR formation and in test SMLC. Further evidence for SEPIR being an immunologically specific product is documented by the finding that spleen cells from mice made specifically tolerant at birth cannot respond to the tolerogen with SEPIR formation, but are fully reactive towards unrelated H-2 antigens. No responses to syngeneic or autologous antigens could be discerned. Characterization of allorecognition resulting in SEPIR formation has revealed that spleen cells of Lyt 23 phenotype recognize H-2K/D-encoded antigens. Lyt 1 cells appear to be incapable of forming the early product and H-2I-determined antigens fail to incite a response. Reactivity to class I antigens was amenable to specific inhibition by monoclonal alloantibodies.
Dynamics of Physiological Changes in Bovine Myofibers during Growth and Sexual Development
Authors: Sink J.D. , Turgut H. , Mann O.M.
Published: October 2008

Semitendinosus (ST) muscle samples were excised from 8 intact and 8 castrate male animals (Bos taurus) when they reached age end-points of 8, 12, 16, and 20 months. All three principal myofiber phenotypes (IC, IIA, IIB) increased in size with increasing age, with the IIA (fast-white) fibers usually larger than the other two types. Only at 16 and 20 months were the type II myofibers from intact males consistently larger than that from castrates. The amount of IIA fibers always exceeded that of the other two phenotypes at every age. Myofiber characteristics were more highly correlated with animal age than with either total body weight or total muscle mass. An ontogenetic scheme is proposed to illustrate the dynamic interrelationships of the three ST myofiber phenotypes.
Hapten Binding Assay Using Guinea Pig Macrophages
Authors: Lefkovits H.
Published: October 2008

An assay for testing the binding of haptens to antigen-presenting cells was developed. The test consists of consecutive incubation of macrophages with the tested hapten, followed by incubation with a radioactively labeled standard. If the first hapten occupies a certain number of binding sites, the other one (radioactively labeled standard) reveals reduced binding. The binding assay in the present modification is fully suitable for measuring the binding of water-soluble haptens. It was found that from a panel of tested haptens, DNFB, β-bromostyrol and phenylacetaldehyde exhibit strong binding.
Release of Proteinases by Cultures of Human Cell Lines Derived from Squamous Carcinomas of the Tongue and Larynx
Authors: Baici Antonio , Sträuli Peter
Published: October 2008

Seven human cell lines derived from squamous carcinomas of the tongue and larynx were examined for their ability to produce and secrete proteinases. All cell lines were able to release into the culture medium cysteine proteinase and plasminogen activator-like activities. All lines differed from each other in the amount of enzymes secreted, in the kinetics of the secretion, in the quality of the enzymes produced an in the intracellular pool of these activities. These features constitute potential criteria for classifying the biochemical behavior of the tumor cell lines and for explaining their different ability to invade soft tissues and bone.
Differences in the Tumoricidal Activation of Rat and Mouse Macrophages by Endotoxins
Authors: Jeannin Jean François , Lagadec Patricia , Reisser Danièle , Martin François
Published: October 2008

Conventional and specific pathogen-free rat resident peritoneal macrophages were lytic to tumor cells in the presence of endotoxins even when not elicited or not stimulated in vivo or in vitro. In contrast, conventional mouse resident peritoneal macrophages were not cytolytic in the presence of endotoxins. The induction by endotoxins of rat macrophage-mediated cytolysis was only obtained after the binding of tumor cells by macrophages. Rat resident peritoneal macrophages bound faster and stronger to tumor cells than mouse resident peritoneal macrophages. These differences in binding could explain the species differences in the tumoricidal response to endotoxins.
Response to Ionizing Radiation of Human Bladder Transitional Cell Carcinomas Grown in the Nude Mouse
Authors: Kyriazis Andreas P. , Yagoda Alan , Kyriazis Aikaterini A. , Kereiakes James G. , McCombs, III William B.
Published: October 2008

The sensitivity to ionizing radiation of three human bladder transitional cell carcinomas grown in the nude mouse was investigated at four different radiation levels, 500, 1,000, 2,000 and 4,000 rad. Each tumor line exhibited a response pattern which reflected to a certain degree tumor differentiation and growth rate in the nude mouse. Exposure to 1,000 rad was the minimum amount of radiation required to produce a distinct, although transient, tumor response in all three tumor lines characterized by a growth delay of 3–4 weeks, whereas maximum tumor response was observed at the 4,000 rad radiation level. These studies would permit a better understanding of the behavior of human bladder cancer to ionizing radiation and may further facilitate efforts at identifying effective radiosensitizing agents that may result in maximizing tumor response.
Transglutaminase Activity Increases in HL60 Cells Induced to Differentiate with Retinoic Acid and TPA but not with DMSO
Authors: Maddox Anne-Marie , Haddox Mari K.
Published: October 2008

HL60 cells induced to differentiate into myeloid cells by retinoic acid exhibited a 300-fold increase in transglutaminase (TGase) activity which peaked on day 5. HL60 cells induced to differentiate into monocytes by a phorbol ester tetradecanoylphorbol-12-myristate-13-acetate (TPA) had a greater than 840-fold increase in TGase activity on day 7. In contrast, cells induced to differentiate along the myeloid pathway by dimethyl sulfoxide (DMSO) exhibited no increase in TGase activity. Elevation of TGase activity appears to be characteristic of monocyte differentiation and retinoic acid-induced myeloid differentiation but not of myeloid differentiation in response to DMSO.
Defective Actin Organization in Cultured Skin Fibroblasts from Individuals with Inherited Colon Adenocarcinoma Is Not Restored by Addition of Fibronectin
Authors: Kopelovich Levy , Murray Clifford , Kleinman Hynda K. , Martin George R. , Fukuda Yuh , Rennard Stephen I.
Published: October 2008

Cultured skin fibroblasts from patients with hereditary adenomatosis of the colon and rectum (ACR) have a disorganized cytoskeleton. Since fibronectin has been found to restore normal cytoskeletal and cellular morphology to transformed cells, we investigated the fibronectin pattern in ACR cells. We found that although both fibronectin synthesis and binding to the cell surface were apparently normal, addition of fibronectin did not restore a normal distribution of actin cables to ACR cells. The data suggest that coupling of cell surface-bound fibronectin to the cytoskeleton might be constitutively defective in ACR cells.
Active Cell Movements and Embryonic Cholinesterase in the Postmetamorphic Completion of the Adrenal Morphogenesis in Frogs
Authors: Grassi Milano E. , Arizzi M. , Bracci M.A. , Manelli H.
Published: October 2008

The adrenal gland in Rana esculenta complex, as in other advanced anurans, is not yet in its definitive position at the end of the metamorphosis and reaches it subsequently, before sexual maturity. The displacement takes place by various means, among which active cellular movements prevail. These are demonstrated by the presence of acetylcholinesterase (AChE) in noninnervated cells (embryonic AChE). The disintegration of the connective tissues which delimit the cords of the adrenal gland, the passive transport of cells by the blood vessels and the movements of adjacent renal and mesenchymal cells collaborate with the active movements of the adrenal cells.
The in vitro Effects of Opines and Other Compounds on DNAs Originating from Bacteria, and from Healthy and Tumorous Plant Tissues
Authors: Le Goff Liliane , Beljanski Mirko
Published: October 2008

Purified total DNAs were isolated from oncogenic or nononcogenic Agrobacterium tumefaciens cells as well as from normal and crown gall tissues. Opines (octopine, nopaline, lysopine), plant hormone (auxin IAA) and some carcinogenic compounds were used in order to correlate their effects on in vitro strand separation and synthesis of DNAs with in vivo tumorous cell multiplication. Octopine (or nopaline) induced chain opening of DNAs originating from octopine (or nopaline)-metabolizing bacteria and from same bacteria strain-induced tumorous cells. This phenomenon was measured by the increase in DNA hyperchromicity which is concentration dependent. The tested compounds stimulated the in vitro synthesis of the same DNAs. Under the same conditions, in vitro strand separation and synthesis of healthy plant DNA was not (or only slightly) enhanced, except in the case of particular hormone-connected healthy cell DNA. IAA and carcinogens stimulated in vitro synthesis and induced in vitro strand separation (dose-dependent effect) of DNAs isolated from crown gall cells and inducing bacteria. Compared to healthy cell DNAs, these DNAs were thus susceptible to structurally very diversified molecules and in this way behave as do mammalian tissue DNAs. The opine and IAA actions observed here were specific for plant tissue DNA; cancerous human or animal tissue DNAs were insensitive. By their presence in the crown gall cells, opines possibly maintain destabilized areas (required for rapid growth and division) on tumor cell DNA. The cooperative actions of IAA and opines as well as small RNA and RNA fragments on gene activation, might explain the autonomy of plant tumor cells.
In vivo Effects of DL-α-Difluoromethylornithine on the Polyamine and Nucleotide Phosphate Metabolism in P388/S Leukemia Cells
Authors: Kremmer T. , Boldizsár M. , Holczinger L.
Published: October 2008

In vivo effects of DL-α-difluoromethylornithine (DFMO) on the metabolism of polyamines and nucleotide phosphates were monitored in P388/S leukemia cells grown intra-peritoneally in BDF1 inbred male mice. Inhibiting the ornithine decarboxylase (ODC) activity DFMO depleted putrescine and spermidine to 30–50 and 50–60%, respectively, and increased spermine to 25–60% compared with the controls, when given as 2% solution in drinking water of the tumor-bearing animals. DFMO treatment caused a parallel 56% elevation of total nucleotide content in tumor cells with distinct and significant increase of some nucleotide phosphates. The most pronounced alterations were shown in the intracellular UTP (202%), CTP (103%), ADP (92%) and ATP (71%) concentrations. Changes in polyamine and nucleotide phosphate metabolisms were dependent on tumor progression. A possible explanation of the metabolic events induced by DFMO is discussed.
Use of Donor-Specific T-Cell Lines for Monitoring of Human Allograft Recipients
Authors: Huber Christoph , Irschik Evelyn , Leiter Evi , Binz Hans , Niederwieser Dietger , Spielberger Martin , Kathrein Hermann , Schönitzer Dieter , Margreiter Raimund
Published: October 2008

Donor-specific and highly cytotoxic T-cell lines (TCL) as well as lectin-induced TCL were established from pretransplant lymphocytes of 6 cadaveric renal allograft recipients. These TCL were used in the 125I-staphylococcus protein A assay to detect IgG antibodies in pre- and posttransplant sera of these patients preferentially binding to autologous donor-specific TCL. Such antibodies were detected in pretransplant sera from 4 of these 6 allograft recipients. Antibody levels in these 4 patients and in 1 additional case who became positive after transplantation further increased during acute cellular rejection episodes. They disappeared after successful treatment but remained elevated until transplantectomy for treatment of irreversible rejection in 1 case. IgG antibodies binding to autologous lectin-induced TCL were detected in only 1 patient and exhibited a pattern clearly different from those binding to donor-reactive TCL. Although attempts to define the antigenic specificity of the autoantibodies binding to donor-specific TCL by genetical and biochemical means has remained unsuccessful so far, the demonstration of their relationship to in vivo expansion of donor-reactive immune cells deserves further attention.
Human Lactic Dehydrogenase as a Marker for Monitoring the Growth of Prostatic Carcinoma in Nude Mice
Authors: Heckl W. , Fogh J.
Published: October 2008

Human lactate dehydrogenase (1,1,1,27-LDH) isoenzymes were determined in human prostatic carcinoma cell lines. The isoenzymes LDH-4/LDH-5 were identified and quantified in the serum of athymic mice bearing the human prostatic carcinoma cell lines PC-3, PC-3/M and DU 145 subcutaneously. The amounts of the slowly migrating isoenzymes released by the solid tumors correlated with increasing tumor volume and tumor weight. These correlations indicate that the human LDH-4/LDH-5 isoenzymes are useful parameters for monitoring the presence and growth of prostatic cancer in the athymic animal model.
Influence of Deciliation and Ciliary Regeneration on Hormonal Imprinting in Tetrahymena
Authors: Csaba G. , Darvas Zsuzsa , Kovács P. , Madarász B.
Published: October 2008

Imprinting induced in Tetrahymena with insulin is not abolished by deciliation. No imprinting occurred in deciliated cells exposed to insulin at 1 or 2 h of regeneration. However, imprinting did occur if Tetrahymena was exposed to insulin after 3 h of regeneration. It appears that while presence of cilia is a prerequisite of imprinting, the pertinent information is not, or not exclusively stored in the cilia.
Comparison of Exogenous Growth Stimuli for Chemically Transformed Cells: Growth Factors, Serum and Cocultures
Authors: Holzer Claudia , Maier Peter , Zbinden Gerhard
Published: October 2008

Rat tumor cells isolated from 2 fibrosarcomas which differed in their degree of differentiation were exposed to growth-stimulating influences in soft agar. The effects of growth factors (epidermal growth factor, fibrosarcoma growth factor and insulin), of fetal calf serum and of cocultured normal mesenchymal cells of rats and nude mice were compared. Each of the 3 growth factors exerted a specific response and dose dependence. Increasing concentrations of serum stimulated the number of cells which formed clones in soft agar. Experiments using combinations of growth factors and fetal calf serum demonstrated that a complex optimal mixture of growth stimuli was responsible for the efficient growth-promoting activity of the serum. In cocultures with normal cells, cloning efficiency of tumor cells was enhanced and growth of tumor cells was accelerated. This stimulus was due to the constant release of an agar-diffusible growth-stimulating factor by the normal, nondividing cells. Cocultured mouse cells showed an even higher growth-stimulating activity than rat fibroblasts. Cells obtained from the poorly differentiated fibrosarcoma responded, in relative terms, better to all growth-stimulating influences, than those derived from the well-differentiated tumor.
Potentiated Susceptibility of Ascites Tumor to Acyl Derivatives of Ascorbate Caused by Balanced Hydrophobicity in the Molecule
Authors: Miwa N. , Yamazaki H.
Published: October 2008

Orders of susceptibility of Ehrlich ascites tumor to L-ascorbic acid (Asc), its 6-stearoyl (6S), 6-palmitoyl (6P) and 2,6-dipalmitoyl (DP) derivatives were assessed in vitro and in vivo: 6P (a 50% growth inhibitory concentration (IC50) for cultured cells, 12 µM; an increased life-span of treated mice, 283%) >6S (61 µM; 240%)& >&;>Asc (430 µM; 122%) ≧DP (>200 µM; 89%), indicating that the enhanced susceptibility was due to acyl moiety substituted at C6-hydroxyl group of Asc, but was retracted by further substitution at C2-hydroxyl group. Equimolar mixture of Asc and palmitic acid, stearic acid or their methyl esters was much less cytotoxic than 6P or 6S. Thus the enhanced susceptibility was not primarily due to an additive cytotoxic effect of ascorbyl and acyl moieties, but to a balanced hydrophobicity introduced into the molecule by a poorly cytotoxic acyl moiety.
Murine Lymphocyte and Embryonal Carcinoma Cell Surface Antigens Recognised by Rabbit Anti-Murine Embryonal Carcinoma Serum
Authors: Stern Peter L. , Beresford Nicola A. , Bell Susan M. , Thompson Stephen
Published: October 2008

Some recent studies have suggested that murine embryonal carcinoma (EC) cells which lack classical H-2 molecules must express some novel class-I-MHC (major histocompatibility complex) type antigens. We have investigated whether rabbit anti-EC sera may recognize such components. Molecules of 40 and 48 kd were immunoprecipitated from EC cells and lymphocytes. The possibility that these molecules may be coded for by genes in the Qa-Tla locus which contains many MHC class-I genes or pseudogenes was investigated. The 40 and 48 kd molecules were not associated with β2-microglobulin on EC cells, nor with each other or other molecules through S-S bonds, although they appeared to have intradisulphide structures. Peptide map analysis suggested that the lymphocyte 40 and 48 kd molecules were related to the EC cell 40 and 48 kd molecules. However, these molecules were different from H-2, la or Qa-2 molecules from the same mouse strain.
Description of and Treatment to Inhibit the Rejection of Human Split-Thickness Skin Grafts by Congenitally Athymic (Nude) Rats
Authors: Gilhar Amos , Wojciechowski Zbigniew J. , Piepkorn Michael W. , Spangrude Gerald J. , Roberts Lee K. , Krueger Gerald G.
Published: October 2008

Gradual rejection of topically engrafted human split-thickness skin grafts (HSTSG) occurred in greater than 90% of congenitally athymic (nude) rats between 21 and 42 days of grafting. Engraftment and rejection of HSTSG is accompanied by a partial restoration of some cell-mediated immune components, the mixed lymphocyte response and lysis of human target cells. Histologic features of the rejection process were those seen in a host-versus-graft reaction. Immunofluorescent analysis of skin undergoing rejection demonstrated IgG at the basement membrane zone in most grafts. Nude rats rejecting HSTSG had circulating IgG which bound to the basement membrane zone and blood vessels of human skin. Nude rats treated with cyclosporine injections for 21 days had an enhanced survival of HSTSG, 120 or more days.
Studies into Disturbing Receptor ‘Memory’ in a Unicellular (Tetrahymena) Model System: Changes in the Imprinting Potential on Exposure to Combinations of Related and Unrelated Hormones
Authors: Csaba G. , Kovács P.
Published: October 2008

Signal molecules (hormones) can induce a hormonal imprinting in the unicellular Tetrahymena, as judged from an increase in binding capacity on reexposure in the subsequent generations. Structurally unrelated polypeptide hormones (insulin, FSH) neutralize each other’s effect, whereas related hormones (TSH-FSH) may amplify or depress it. The non-signal polypeptide molecule BSA prevents hormonal imprinting on combined exposure. The glucocorticoid hormone enhances binding capacity for insulin (as in mammals) and increases its imprinting potential, whereas on TSH and FSH it has no similar effects. The amino acid hormone serotonin increases the binding capacity for TSH and FSH in non-pretreated cells, but depresses rather than stimulates the imprinting potential of these hormones, measured in the case of the second encounter.
Expression of Epidermal Differentiation Antigens in Cultures of Interspecific Somatic Hybrids (Human Keratinocytes × Mouse Fibroblasts 3T3–4E)
Authors: Lizard Gerard , Lery Xavier , Chardonnet Yvette , Viac Jacqueline , Fargier Marie-Claire , Thivolet Jean
Published: October 2008

Interspecific somatic hybrids have been prepared by fusion of human epidermal cells with mouse fibroblasts 3T3–4E using PEG 4000. Expression of epidermal differentiation antigens (bullous pemphigoid antigens, BP, keratin subsets 55–57 k and 67 k), markers of basal and suprabasal cells, were studied by immunocytochemistry for 10 passages. These markers were detected in the hybrids early after fusion, indicating that cells from both compartments were able to fuse with 3T3–4E cells. However, the hybrids expressing high molecular weight keratins were no longer detected after 7 days in primary cultures and serial passages, whereas those expressing BP antigens and vimentin persisted. Low molecular weight keratins 52 K and 50 K were detected by SDS-PAGE at the second passage in precipitates formed between labeled hybrid lysates and total keratin rabbit antiserum. Karyo-type analysis showed mainly murine chromosomes and a submetacentric human chromosome between the 6th and the 10th passage.
Alterations of MCF-7 Human Breast Cancer Cell after Prostaglandins PGA1 and PGF2α‘Treatment
Authors: Shahabi N.A. , Chegini N. , Wittliff J.L.
Published: October 2008

Treatment of monolayer cultures of MCF-7 cells with prostaglandins PGA1 and PGF2α inhibited cell proliferation, reduced the rate of labeled precursor incorporation into DNA, RNA, and protein, and induced morphological changes in a dose-dependent manner. The rate of [3H]thymidine incorporation was increased by PGA1 at l0–10–l0–8M, while a sharp decrease was observed at 10–6–10–4 M(p < 0.05 and p < 0.005). PGF2α inhibited [3H]thymidine incorporation at all concentrations tested. Similar results were obtained for [3H]uridine incorporation with both PGs. PGA1 inhibited [3H]leucine incorporation at 10–4M, but increased incorportion at 10–10–10–6M. At the ultrastructural level, neither PG induced morphological alterations at 10–12–10–8M. However, at 10–6–10–4 Mboth PGA1 and PGF2α diminished the number and size of cell surface projections; some cells appeared to completely lack microvilli. Disorganization of mitochondrial cristae and increased electron density of the matrix were also evident
Guanine Salvage by Organ Cultures of Mouse Tooth Germs
Authors: Dye Frank J.
Published: October 2008

The effect of mycophenolic acid (MPA) which inhibits the biosynthesis of guanosine monophosphate (GMP) in organ cultures of mouse tooth germs can be partially counteracted by adding guanine to the MPA cultures. This may be due (1) to salvaging guanine by the enzyme hypoxanthine guanine phosphoribosyl transferase (HGPRT), or (2) to competition for a common membrane carrier involved in mediated transport of both guanine and hypoxanthine in normal biosynthesis and also of MPA. Experiments were carried out to compare the effect of either hypoxanthine or guanine on the MPA-caused inhibition. While addition of guanine to the MPA cultures (MPAG) supports growth equal to controls and development of dental-enamel junction (DEJ) to a level intermediate between control and MPA the addition of hypoxanthine (MPAHX) supports growth and DEJ development not better than MPA. This indicates that guanine is salvaged by HGPRT to GMP while hypoxanthine, salvaged to inosinic acid (inosinic monophosphate, IMP) is ineffective because the MPA inhibition is on the pathway from IMP to GMP.
Protein Synthesis in Heterotopically Transplanted Rat Hearts
Authors: Currie William , Sharma Vijay K. , Stepkowski Stanislas M. , Payce Ronald F.
Published: October 2008

This study examines protein synthesis in heterotopically transplanted rat hearts and several tissues of recipient rats. Donor hearts and recipient tissues synthesized many of the normally occurring proteins observed in tissues of unstressed rats. In addition, a stress-induced protein with a molecular mass of 71 kilodaltons was synthesized in donor heart, recipient heart and lung. Donor hearts incorporated more L-[35S]-methionine than did recipient hearts. Tissues of recipient rats also incorporated more label than the respective tissues of sham-recipient rats. These results suggest that ischemia, endured by the donor hearts during transplantation, induced these changes in protein synthesis.
Characterization and Expression Kinetics of an Endothelial Cell Activation Antigen Present in vivo Only in Acute Inflammatory Tissues
Authors: Goerdt Sergij , Zwadlo Gabriele , Schlegel Renate , Hagemeier Hans-Hermann , Sorg Clemens
Published: October 2008

Endothelial cell activation by endotoxin (LPS), tumor necrosis factor (TNF), Interleukin-1-α, β (IL-1-α β) and phorbolesters (TPA) results in increased monocyte adhesion. Examination of kinetics of monocyte adhesion shows that the onset of adherence enhancement (AE) is similar in all five agents (about 300% AE at 6 h), while its decrease is delayed in LPS/TNF versus IL-1-α, β /TPA-induced activation (LPS versus IL-1- β:260% versus 60% at 18 h). Monoclonal antibody (4D10), raised against 24 h LPS-stimulated endothelial cells detects an endothelial cell-specific activation antigen at Mr 81,000 that is induced by LPS, TNF, IL-1-α β and TPA (within 6 h about 100% positive cells). Decrease in antigen-positive cells is delayed in LPS/TNF versus IL-1-α, β /TPA-induced antigen expression (LPS vs. IL-1-β: 60% vs. 5% at 24 h). In situ the antigen is not expressed in normal and chronic inflammatory tissues. Acute inflammatory tissues, including contact and atopic dermatitis, psoriasis and periodontitis, however, show endothelial cells staining strongly positive. In contact eczemas at different times after elicitation (0, 6, 24, 72, 96 h), expression of the antigen is first seen after 24 h and is still strong at 96 h. These data indicate that LPS/TNF conduct an endothelial cell activation program in vitro, showing the same prolonged kinetics that is found for endothelial cell activation in the acute inflammatory process in vivo.
Plasma Membrane-Associated Cysteine Proteinases in Human and Animal Tumors
Authors: Sloane Bonnie F. , Rozhin Jurij , Hatfield James S. , Crissman John D. , Honn Kenneth V.
Published: October 2008

The ability of tumor cells to invade into and through normal tissue during the metastatic cascade has been attributed to tumor-associated degradative enzymes including proteinases of the metallo, serine and cysteine classes. Work from several laboratories has established that the cysteine proteinases cathepsins L and B are released from tumor cells, primarily as latent precursor forms. In addition, a cathepsin B-like cysteine proteinase has been shown to be associated with the plasma membrane fraction of several animal and human tumors. This form of the enzyme retains activity under physiologic (or pathologic) conditions including at neutral pH and in the presence of low Mr inhibitors. Since we have established that cathepsin B can degrade the basement membrane attachment glycoprotein laminin, we speculate that plasma membrane-associated cathepsin B may participate in focal dissolution of the basement membrane during tumor cell extravasation.
Differences in Growth Factor Sensitivity between Primary and Transformed Murine Cell Cultures Revealed by BrdU/Hoechst Flow Cytometry
Authors: Kubbies M. , Hoehn H. , Rabinovitch P.S.
Published: October 2008

Spontaneous cell transformation is a common feature of all murine cell cultures grown in vitro for extended periods of time. During early passages, these cultures show either progressively reduced growth or complete cessation of growth; after such a ‘crisis’ they display increasing growth rates and unlimited lifespan. Here we use a novel bromodeoxyuridine/Hoechst flow-cytometric technique to examine the growth response of diploid and transformed cells of the murine species Micromys minutus under a variety of growth conditions. After spontaneous transformation, growth factor exposure results in increased G0/Gl cell recruitment and higher growth rates than shown by the nontransformed diploid cell fraction. Despite clonal heterogeneity, this difference is seen at all fetal calf serum (FCS) concentrations, although it is most pronounced with low serum. Epidermal growth factor and insulin are shown to act synergistically and promote growth equal to exposures of transformed cultures to 10% FCS. The observed differences in growth factor response between diploid and aneuploid cells could explain the reported lack of a classical growth crisis in growth factor-supplemented media during the spontaneous in vitro transformation of primary murine cell cultures.
Human Monoclonal Antibodies Derived from Pleural Effusion Lymphocytes of a Patient with Breast Carcinoma React with Human Breast Cancer-Associated Antigens and Mouse Mammary Tumor Virus Polypeptides
Authors: Smorodinsky Nechama I. , Argov Shmuel , Ghendles Yosi , Bakimer Ronit , Krup Margalith , Lupu Ruth , Keydar Iafa , Shoenfeld Yehuda
Published: October 2008

Four human hybridoma cell lines (PEB1-4) were established from a fusion of pleural effusion lymphocytes isolated from a breast cancer patient with metastatic disease, 6 years postmastectomy. The hybridomas secreted IgG-k (3 µg/ml/l06 cells). These monoclonal antibodies (PEB1-4) reacted to different degrees with mouse mammary tumor virus (MMTV) and T47D particles (HuMTV). Immunological cross-reaction was also detected with antigens isolated from body fluids of breast cancer patients (BF-Ag). The binding capacity of the monoclonal antibodies (MAbs) PEB1-4 to the above-mentioned antigens was measured by RIA. The specificity of these antibodies was further demonstrated by radioimmunoprecipitation of MMTV, T47D (HuMTV) and BF-Ag. The binding of PEB1-4 to surface antigens of intact cells grown in culture was measured by RIA. Some of the MAbs were shown to bind more avidly to breast cancer cells than to nonbreast cancer cells or nonmal-ignant cells. The PEB1-4 human monoclonal antibodies may be found useful in analyzing the virus-breast cancer relationship.
Fibroblasts from Persons Predisposed to Cancer Are Abnormally Resistant to 5-Azacytidine-Induced Cytotoxicity
Authors: Kopelovich Levy , Fenyk Judith
Published: October 2008

We have determined the sensitivity to 5-azacytidine of cultured fibroblasts obtained from clinically defined areas on the skin of patients with hereditary adenomatosis of the colon and rectum (ACR) and von Recklinghausen neurofibromatosis (NF). Fibroblasts from normal appearing skin of both ACR and NF patients were about 2- to 3-fold more resistant to 5-azacytidine-induced cytotoxicity than were fibroblasts obtained from comparable areas of normal persons. Fibroblasts from café-au-lait lesions and from neurofibromas (NF) were increasingly more resistant to 5-azacytidine than were fibroblasts taken from normal appearing skin of the same NF patient as well as of patients from different pedigrees. The results show that fibroblasts from persons predisposed to cancer and from cancer-prone tissues in such persons are abnormally resistant to 5-azacytidine as determined by the cloning efficiency assay.
Contribution of the Extracellular Matrix to Growth Properties of Cells from a Preneoplastic Outgrowth: Possible Role of Hyaluronic Acid
Authors: Elstad Catherine A. , Hosick Howard L.
Published: October 2008

Hyaluronic acid (HA) accumulates around actively growing normal and tumorigenic mammary epithelial cells and has been implicated as a modulator of cell proliferation. We have tested the role of exogenous HA presented in several different forms in in vitro growth regulation of a cell line (CL-S1) derived from preneoplastic mouse mammary tissue. This cell line grows slowly and synthesizes very little HA. We first assessed growth of CL-S1 cells seeded onto actual matrix generated by CL-S1 cells themselves (which has a low HA content) or by a related tumorigenic cell line, +SA, that generates an HA-rich matrix. Growth on both these HA-containing substrata was significantly enhanced above control values on plastic. Growth on the +SA biomatrix was over 5 times greater than on tissue culture plastic and significantly greater than that seen with all other treatments. Differences in growth responses of CL-S1 cells seeded atop CL-S1- and +SA-derived matrices could be attributable to differences in matrix HA content. As a more direct test of this possibility, growth responses of CL-S1 cells to HA covalently bonded to tissue culture dishes and to HA dissolved in culture media were tested. Growth on the prepared HA substrata was consistently twice that on plastic. In soluble form, HA at a concentration of 100 µg HA/ml culture medium, stimulated CL-S1 growth 196 and 125% of control in monolayer cultures, respectively, seeded at low (≈ 102 viable cells/cm2) and high (≈ 104 viable cells/cm2) densities on plastic. Higher HA concentrations inhibited growth at low seeding densities. Comparable responses were noted for cells grown on collagen gel substrata. Our results suggest that accumulation of HA extracellularly can affect growth properties of cells from preneoplastic lesions.
Modulation of Tumor Cell Motility by Prostaglandins and Inhibitors of Prostaglandin Synthesis
Authors: He Xuanmin , Fligiel Suzanne E.G. , Varani James
Published: October 2008

15-Methyl-prostaglandin E1 (15-M-PGE1), a synthetic stable, prostaglandin E1 analogue was examined for ability to inhibit motility in a line of murine tumor cells. Inhibition of random motility and motility stimulated by 12-O-tetradecanoyl phorbol acetate was seen at concentrations of 15-M-PGE1 as low as 1 µM. Inhibition of laminin-stimulated motility was observed with 10 µM 15-M-PGE1. The murine tumor cells used in this study produced high levels of prostaglandins. When the cells were treated with either indomethacin or ibuprofen, prostaglandin levels (measured as prostaglandin E2 by radioimmunoassay) were reduced by greater than 95% without a corresponding increase in lipoxygenase products. When indomethacin or ibuprofen-treated cells were compared to control cells in regards to motility, they were more active. These studies show that E-series prostaglandins can modulate motility in the murine fibrosarcoma cells and suggest that the production of endogenous cyclooxygenase metabolites by the murine tumor cells may regulate, at least in part, the responsiveness of the cells to stimulation.
Interaction between Cisplatin-Treated Macrophages and Dalton’s Lymphoma Cells in vitro
Authors: Singh Sukh Mahendra , Sodhi Ajit
Published: October 2008

Murine peritoneal macrophages treated with cisplatin (10 µg/ml) showed increased binding to Dalton’s lymphoma cells in vitro. Macrophages and target cells both extend cytoplasmic extensions towards each other, which finally join and fuse to form a distinct cytoplasmic bridge between the two cells. At later stages of coincubation the macrophages and tumor cells get closely bound with several short cytoplasmic connections. Finally the plasmalemmae between the two cells fuse over a large surface area and the tumor cell is phagocyted. No tumor cell was found to form cytoplasmic bridges when incubated with untreated macrophages. The base of cytoplasmic bridge and the cytoplasmic bridge between the macrophage and tumor cells stained for actin and fibronectin, but not for tubulin. We also report the transfer of lysosomes from the cytoplasm of cisplatin-treated macrophages to the tumor cell cytoplasm through cytoplasmic bridges. It is further reported that cisplatin-induced macrophage cytotoxicity against DL cells is inhibited by nifedipine and chlorpromazine.
Phorbol Ester Receptors and the Induction of Differentiation in the Human T Lymphoblastic Cell Line MOLT-3
Authors: Mayumi Takehito , Nagasawa Kohei , Horiuchi Takahiko , Kusaba Tomohiro
Published: October 2008

To examine the mechanism of induction of differentiation in the human malignant T-lymphoblastic cell line, MOLT-3, by 12-O-tetradecanoylphorbol-13-acetate (TPA), the role of receptors for phorbol esters was investigated. Binding of [20–3H]-phorbol-12,13-dibutyrate to TPA-resistant subclones derived from MOLT-3 was less than50 % of that of the parental MOLT-3. Scatchard analysis showed that the concentration of phorbol ester receptors in a TPA-resistant subclone was about 50% of that in the parental MOLT-3, but affinities of binding were similar, indicating that more than a certain number of phorbol ester receptors is required to induce differentiation by TPA in this human T cell line.
Human Trophoblast Glycoproteins Defined by Monoclonal Antibody 1D2
Authors: Hole Nicholas , Barton-Hanson Nicholas , Berwick Steven , Stern Peter
Published: October 2008

A cell surface antigen has been defined by a monoclonal antibody 1D2, raised following immunisation with lectin-purified syncytiotrophoblast glycoproteins. 1D2 was nonreactive with any one of 8 common trophoblast proteins in immunodot. Analysis of nonreduced western blots of syncytiotrophoblast microvillous plasma membrane (StMPM) protein indicated that mAb 1D2 was reactive with a series of sialylated proteins with molecular weights of 16–22 kilodaltons. Immunoprecipitates of radiolabelled StMPM protein contained molecules that co-migrated with placental alkaline phosphatase in addition to those identified by western blotting. This set of human trophoblast molecules has not been previously identified by monoclonal antibodies; the antigenicity is widely distributed in human tissues.
Characteristics of Cyclic AMP Enhancement of Retinoic Acid Induction of Increased Transglutaminase Activity in HL60 Cells
Authors: Maddox Anne-Marie , Haddox Man K.
Published: October 2008

When the human myeloid leukemia cell line (HL60) is induced to differentiate with retinoic acid (RA), there is a concentration-dependent increase in transglutaminase (TGase) activity which peaks on day 5. While dibutyryl 3’,5’-cyclic adenosine monophosphate (db-cAMP) alone produced only a slight increase in TGase activity in HL60 cells, the con-comittant addition of db-cAMP (100 µM) with RA (10–12–10–4M) potentiates RA induction of TGase activity. Maximal increases in TGase activity (2- to 10-fold) were observed with 10–4–10–7M RA and when db-cAMP was present from 24 to 48 h after the addition of RA. The cyclic nucleotide enhancement was dose-dependent from 10 to 100 µM of cAMP. Less marked increases were observed with 8-bromo-cAMP and with the phosphodiesterase inhibitor theophylline. Although the simultaneous addition of PGE1 or PGE2 (10-8-10-6M) produced no enhancement of RA-induced TGase activity, adding PGE1 or PGE2 24 or 48 h following RA treatments produced an enhancement of TGase activity. The phosphodiesterase inhibitor potentiated the increases produced by db-cAMP and the prostaglandins. Dibutyryl cAMP enhanced the ability of RA to induce the cells to reduce nitroblue tetrazolium (NBT), a functional measure of differentiation, at lower concentrations of RA and with shorter treatment durations. cAMP potentiates RA-induced TGase activity in HL60 cells and the combination appears to be associated with enhanced RA-induced differentiation.
Extracellular Materials and Determination of Neuroectoblast in Amphibian Gastrula
Authors: Sánchez Sara S. , Barbieri Francisco D.
Published: October 2008

The biological activity of the extracellular material (ECM) located in the dorsal and ventral regions of Bufo arenarum is assayed. Ectoblast cells were isolated and cultured with ECM taken from the embryonal regions. The epiblastic cells treated with dorsal ECM, mainly located at the interphase between the invaginating blastoporal lip and the overlying ectoblast differentiated morphologically into neural, mesenchyme and pigment cells. In contrast, control epiplastic cells, cultured either in salt solution or in ventral region ECM, differentiated into ciliated and secretory cell types. The results reported here provide evidence that the dorsal region ECM of the embryo can induce neural differentiation in indeterminate epiblastic cells.
Down-Regulation of Murine Collagen-Induced Arthritis by a T Cell Hybridoma
Authors: Kresina Thomas F.
Published: October 2008

T cell hybridoma cell lines were generated by somatic cell fusion of BW 5147 myeloma cells and splenic cells from mice suppressed for collagen-induced arthritis (CIA). Two cell lines were characterized for their cell surface phenotype, antigen recognition and capacity to down-regulate the erythema and edema associated with CIA. Cell line T101N was determined to portray the cell surface phenotype Lyl+2- L3T4- Thyl+ by a direct binding assay. Cell line T104B1 was determined to express only the Thyl + alloantigen. Panning studies, measurement of IL-2 production in vitro and the suppression of antibodies to type I and type II collagen in vivo indicate that the hybridoma cells are not isotype specific in their recognition of the polymorphic interstitial collagens. Down-regulation of the erythema and edema of CIA occurred on injection of 1 × 105 T101N cells but not T104B1 cells. Histology of the tarsus region of the hind paw of CIA mice 33 days after the administration of T101N cells showed contrasting histopathology compared to that of CIA mice. The joints of CIA mice given T101N cells showed aligned articular surfaces resembling normal joint structure and only residual pannus. The data indicate that collagen-specific cloned T cell lines can modulate the gross pathology and joint architecture of joints exhibiting CIA.
Origin of Cells in Human Oral Mucosa Heterotransplants in Nude Mice
Authors: Holmstrup Palle , Gram Niels
Published: October 2008

Species identification of cells was performed in a model to study epithelial-mesenchymal interactions using combinations of human and murine tissue. The study comprised 34 successfully recovered transplants of human palatal mucosa with 15 human epithelial outgrowths formed on uncertainly species-identified connective tissue. Formalin-fixed and paraffin-embedded 5-µm sections were stained with bisbenzimide (0.8 µg/ml) and examined in an epifluorescence microscope. The nuclear-staining pattern revealed limited peripheral invasion of the transplanted connective tissue by murine fibroblasts. In areas of epithelial outgrowth a small number of fibroblasts could not be identified but most subepithelial fibroblasts were murine. The study supports previous findings which indicate that the altered differentiation of the human epithelial outgrowths was caused by the murine connective tissue underlying the human epithelium.
Ultrastructural and Autoradiographic Investigations of Cell Cultures Derived from Tendons or Ligamentous Material from Patients with Fibromatous Disorders
Authors: Neumüller Josef , Tohidast-Akrad Makyieh , Ammer Kurt , Hakimzadeh Ahmad , Stransky Gregor , Weis Serge , Partsch Gerald , Eberl Rudolf
Published: October 2008

Cell cultures were derived from tendons or ligamentous material from patients with carpal tunnel syndrome (CTS), Dupuytren’s contracture (DP), tendopathia nodosa (TN) and hallux valgus (HV). The ultrastructure of the operation specimens as well as of the cell monolayers was investigated, using a floating sheet method in order to preserve both cell-to-cell contacts and the orientation of the monolayers. The histologic features of the tissues obtained in the operations were correlated with the ultrastructure of the cells in culture derived from these specimens. In DP, above all in the nodules, an activation of the capillary endothelium in the vicinity of myofibroblasts and mast cells was observed. In CTS the collagen fibrils varied extremely in diameter. In DP and TN biopsies a splicing process of helicoidly arranged fibrils could be seen. A disintegration of elastic fibers in the fibrillar and amorphous components was found in DP nodules, HV and TN tissues. Transitional forms between fibroblasts and myofibroblasts were observed not only in DP but also – though in a smaller percentage – in the cultures derived from the other patients. The cells showed organelles for active protein synthesis and transport. Autophagocytosis and the formation of multilamellated bodies took place in TN and HV cultures. In CTS, DP and TN cultures cells were connected via gap junctions. In some cultures, above all in those derived from CTS, monocilia were found. In CTS cultures the formation of intracellular collagen occurred. Growth parameters were rather low in HV cultures. PLmax (maximal pulse labelling index) values were higher in TN cultures than in DP and HV cultures. Plating efficiency (PE) values were higher in cultures derived from cell-rich and capillarized tissues than in biopsies with few cells.
Adrenocorticotropin Binding Activity of B16/C3 Melanoma Cells
Authors: Simić-Krstić Jovana , Ševaljević Ljiljana
Published: October 2008

The binding of adrenocorticotropic hormone (ACTH) to B16/C3 murine melanoma cells was found to be specific and saturable. The binding capacity of the cells changed as a function of the age of the culture. Scatchard analysis revealed one class of high-affinity ACTH binding sites. The specificity of ACTH binding to the cells was tested by displacement experiments with human leukocyte interferon (α-IFN) and α-melanocyte stimulating hormone (α-MSH) as the competitors. Structure-activity relationship of ACTH, α-MSH and α-IFN was discussed.
Correlation of Anionic Residue Mobility at the Filopodial Plasma Membranes with Multilayer Formation in Transformed 3T3 Cells
Authors: Kawai Norio , Kawai Makoto , Nishimura Toshikazu , Arimura Kazuto , Ichihara Ichiro
Published: October 2008

Simian virus 40-transformed 3T3 (SV40–3T3) cells formed multilayers on a Falcon dish and had numerous filopodial projections, some of which intertwined with those of adjacent cells, in contrast to the few projections of their nontransformed counterparts. When these cells were incubated with polycationic ferritin (0.5 mg/ml), ferritin particles, representative of anionic sites, were spread widely on their surfaces at 4° C, while they formed clusters at 37°C, especially on filopodial surface areas opposing adjacent projections in SV40–3T3 cells. These findings demonstrate an increase in the mobility of molecules with anionic residues on filopodial plasma membranes in SV40–3T3 cells, thus suggesting a role for these projections in the formation of multilayered cell aggregates.
Differentiation of Mononuclear Blood Cells into Macrophages, Fibroblasts and Endothelial Cells in Thrombus Organization
Authors: Leu H.J. , Feigl W. , Susani M. , Odermatt B.
Published: October 2008

The organization of thrombi and emboli may be performed exclusively by mononuclear blood cells which represent precursors of various mesenchymal cell types. Between the 10th and 20th day after the onset of blood clotting, mononuclear cells within the fibrinous matrix differentiate into (1) macrophages responsible for hematoclasia and hematophagocytosis, (2) endothelial cells lining autolytic slits in the fibrinous matrix and forming new capillaries, and (3) fibroblasts and even smooth muscle cells building up a young mesenchymal connective tissue.
Inhibition of Mammary Carcinoma Growth by Retinoidal Benzoic Acid Derivatives
Authors: Fontana Joseph A. , Hobbs Peter D. , Dawson Marcia I.
Published: October 2008

The growth of many breast carcinoma cell lines is inhibited by vitamin A, and derivatives as well as synthetic retinoids. New retinoidal derivatives have recently been synthesized. These retinoidal benzoic acid derivatives displayed enhanced potency in their ability to reverse hamster tracheal keratinization and inhibit ornithine decarboxylase induction in mouse epidermis. We therefore screened a series of analogues of these compounds for their ability to inhibit human breast carcinoma cell proliferation utilizing three estrogen receptor-positive and two estrogen receptor-negative cell lines. The compound (E)-4–2-(5,6,7,8)tetrahydro-5,5,8,8-tetramethyl-2-naphtalenyl)propenyl benzoic acid (Ro 1374–10) was approximately 2–3 orders of magnitude more potent than all-trans-retinoic acid in inhibiting breast carcinoma cell proliferation while the compound SRI-6409–40, which differs from Ro 1374–10 only by the position of a methyl group, was 50-fold more potent than Ro 1374–10. All of the compounds tested displayed were inactive against the estrogen receptor-negative breast carcinoma lines.