Anal Biochem, 1988 May 1, 170(2), 456 - 62 Heparin requirement for the quantitation of fibrinogen production by primary hepatocyte cultures; Dang CV et al.; We have reported a rapid method for the quantitation of proteins secreted in culture media (F.M . LaDuca, C.V . Dang, and W.R . Bell (1986) Anal . Biochem . 158, 262-267) . Using the same method, we observe that serum-free rat hepatocyte cultures exhibited a 100% increase in detectable secreted fibrinogen-antigen in the presence of 1 unit/ml heparin or greater at 24 h of culture . The amount of transferrin, haptoglobin, and albumin detected was unaltered by the presence of heparin . Since heparin is known to affect certain cellular functions, the fates of {35S}methonine-labeled fibrinogen in cell extracts and culture media were examined employing pulse-chase experiments . Labeled intracellular fibrinogen disappeared at similar rates and was initially released into the media in similar amounts in the presence or absence of heparin . At 8 h during the chase, there was a 40-50% reduction in fibrinogen-antigen in spent culture medium lacking heparin . The presence of heparin did not alter the proteolytic degradation of secreted fibrinogen as determined by immunoblotting of spent culture media proteins separated by polyacrylamide gel electrophoresis . In vitro experiments indicate that clotting of fibrinogen by thrombin reduces the amount of immunodetectable fibrinogen . The results indicate that heparin increases the amount of detectable fibrinogen secreted by cultured hepatocytes by preventing clotting and not by stimulating synthesis or secretion or by inhibiting degradation . Hence, it is critically important to include heparin when secreted fibrinogen is quantitated by the method that we have developed. Acta Anaesthesiol Scand, 1988 May, 32(4), 308 - 9 The Venflon cannula as a sideport of infection; Cozanitis DA et al.; The possible role of infection through the valved sideport of Venflon cannulae was evaluated by one individual who followed an identical procedure of placing a syringe into the sideport and injecting saline solution . Basic aseptic techniques were used in a contaminated, busy environment . No bacterial growth was found in either anaerobic or aerobic culture media following a total of 1500 injections . The experiment shows that if the measures used could be applied, infection through the sideport would be minimal. J Endocrinol, 1988 May, 117(2), R5 - 8 Development of a radioimmunoassay for ovine trophoblast protein-1, the antiluteolytic protein from the sheep conceptus; Vallet JL et al.; A radioimmunoassay has been developed for quantitation of ovine trophoblast protein-1 (oTP-1), a sheep conceptus secretory protein which allows for maintenance of the corpus luteum during early pregnancy . The assay was validated for dialysed and undialysed culture medium and pregnant uterine flushings ranging from no dilution (neat) to dilutions of 1:2500 for dialysed media, 1:100-1:1000 for undialysed media and 1:50-1:1000 for pregnant uterine flushings . The assay accurately measured oTP-1 added to undiluted and diluted dialysed and undialysed culture media and pregnant uterine flushings . No cross-reaction was detectable for bovine alpha or gamma interferon, bovine calmodulin, feline conceptus secretory proteins, equine conceptus secretory proteins, porcine conceptus secretory proteins, bovine conceptus secretory proteins and proteins in a uterine flushing collected from a non-pregnant ewe . Immunoreactivity in the assay matched that for oTP-1 throughout oTP-1 purification . This assay is the first validated assay which may be used to quantitate production of oTP-1 in culture or content of oTP-1 in uterine flushings. In Vitro Cell Dev Biol, 1988 May, 24(5), 457 - 63 Characterization of extended primary and secondary cultures of hamster tracheal epithelial cells; Niles R et al.; Studies on the regulation of differentiation in airway epithelial cells have been hampered by the lack of cell culture systems that differentiate in vitro . One such system that does exhibit differentiation is hamster tracheal epithelial cells (HTE) . A major problem with this system, however, is that at the time cells differentiate, they lyze the collagen gel upon which they grow, resulting in termination of the culture . Here we report that by growing the HTE cells at 32 degrees instead of 37 degrees C we can totally prevent lysis of the collagen gel . Cells grown at this lower temperature maintain their differentiated phenotype as evidenced by abundant mucus granules and the secretion of authentic mucus glycoproteins into the culture media . We have also developed a method for subculturing the primary cells which allows growth and differentiation in secondary culture . The HTE cells were capable of being passaged at least three times and did not become transformed as judged by their inability to grow in soft agar and to produce tumors in syngeneic animals . This improved HTE cell culture system will allow detailed studies on the mechanisms which regulate growth, differentiation, and mucus secretion in surface airway epithelial cells. Virology, 1988 May, 164(1), 91 - 8 Binding of cowpea chlorotic mottle virus to cowpea protoplasts and relation of binding to virus entry and infection; Roenhorst JW et al.; Cowpea chlorotic mottle virus (CCMV) and cowpea protoplasts were used to study initial interactions between virus and protoplast . Protoplasts and virus were incubated under varying conditions of temperature, pH, ionic strength, and the presence of added compounds . Both the amount of 35S-labeled virus bound to protoplasts and the percentage of infected cells were determined . At 0 and 25 degrees the amount of virus associated with protoplasts increased with the amount of virus added . With inoculum of 25 x 10(6) virus particles per protoplast, 4 x 10(3) and 14 x 10(3) particles per protoplast were bound at 0 and 25 degrees, respectively . In the presence of polyethylene glycol, 85 x 10(3) associated particles per protoplast were bound at both temperatures and ca . 50% of the protoplasts became infected . No infection occurred in the absence of PEG . Variation of pH or ionic strength in the absence of PEG caused little to no change in binding and no infection . In the presence of PEG, increase of pH resulted in lower binding, but infectivity was not affected . Increasing ionic strength, however, increased both binding and infectivity . The presence of unlabeled CCMV, tobacco mosaic virus coat protein, bovine serum albumin, and polycations during inoculation in the absence of PEG decreased the amount of bound CCMV . In contrast, CCMV coat protein, which has a positively charged N-terminal arm, increased binding . In the presence of PEG the effects were similar, although larger amounts of virus were bound . The percentage of infection was reduced by all additives to 5-25% . Addition of ammonium chloride, which inhibits endocytotic virus uptake in animal cells, during inoculation as well as in culture media, did not reduce infectivity . These data do not support a specific receptor-mediated endocytotic uptake of virus but favor a nonspecific mechanism of entry, possibly through membrane lesions . Observations in the electron microscope support the latter mechanism. Am J Clin Pathol, 1988 May, 89(5), 671 - 4 Evaluation of broth media for routine culture of cerebrospinal and joint fluid specimens; Reinhold CE et al.; Broth cultures of cerebrospinal and joint fluids are important components in the culture detection of meningitis and septic arthritis . The authors examined 121 strains of bacteria isolated from clinical specimens representing 13 species or groups that cause meningitis and arthritis for growth in supplemented Thioglycolate broth (THIO), Supplemented Peptone Broth (SPB), and minced beef heart (MBH) media each alone or with added IsoVitaleX . Both SPB and MBH with IsoVitaleX performed better as broth culture media than the media without IsoVitaleX or THIO with or without IsoVitaleX. J Vasc Surg, 1988 May, 7(5), 697 - 705 Polyglactin 910/polydioxanone bicomponent totally resorbable vascular prostheses; Greisler HP et al.; Previous studies from our laboratory have shown that bioresorbable vascular prostheses woven from lactide-glycolide copolymers and implanted into arteries of several animal models become replaced by cellular tissues; the rate of replacement parallels the kinetics of prosthetic resorption . This study evaluates the efficacy of bicomponent resorbable prostheses as a method of augmenting resistance to dilatation during the resorption period of the more rapidly resorbed component . Bicomponent prostheses (n = 37) were woven from compound yarns containing 74% polyglactin 910 (PG910) and 26% polydioxanone (PDS) and were interposed into adult white New Zealand rabbit infrarenal aortas . Resultant prosthesis-tissue complexes were harvested after 2 weeks to 12 months . Specimens were photographed and sectioned for light, scanning, and transmission electron microscopy . Randomly selected fresh explants at 1 and 3 months and control aortic segments from the same rabbits were simultaneously perfused with culture media (37 degrees C, 100/80 mm Hg, 60 ml/min) and perfusates assayed by means of tritiated radioimmunoassay techniques for the stable prostacyclin metabolite 6-keto-PGF1 alpha before and after the addition of sodium arachidonate (10 micrograms/ml) to the media . Results showed 100% patency, no aneurysms, and stenosis in 1 of 37 prostheses (3%) . PG910 was totally resorbed by 2 months and PDS by 6 months . By 1 month inner capsule thickness was 303 +/- 30 microns . In contrast to previous reports this was significantly thicker than that within 100% PDS (230 +/- 40 microns) and significantly less thick than in 100% PG910 (530 +/- 62 microns) . Inner capsules in all three groups stabilized at similar thicknesses (417 to 502 microns).(ABSTRACT TRUNCATED AT 250 WORDS) Int J Food Microbiol, 1988 May, 6(3), 263 - 8 Inhibitory effects of selected Turkish spices and oregano components on some foodborne fungi; Akgul A et al.; The inhibitory effects of 10 selected Turkish spices, oregano essential oil, thymol and carvacrol towards growth of 9 foodborne fungi were investigated in culture media with pH 3.5 and 5.5 . The antifungal effects of sodium chloride, sorbic acid and sodium benzoate and the combined use of oregano with sodium chloride were also tested under the same conditions for comparison . Of the spices tested, only sodium chloride were also tested under the same conditions for comparison . Of the spices tested, only oregano at 1.0, 1.5, 2.0% (w/v) levels showed effect on all fungi . 8% (w/v) sodium chloride was less effective than oregano . Oregano essential oil, thymol or carvacrol at concentrations of 0.025% and 0.05% completely inhibited the growth of all fungi, showing greater inhibition than sorbic acid at the same concentrations . The combined use of oregano and sodium chloride exhibited a synergistic antifungal effect. In Vitro Cell Dev Biol, 1988 May, 24(5), 401 - 12 A quantitative analysis of lectin binding to adult rat hepatocyte cell surfaces; Jauregui HO et al.; A quantitative evaluation of lectin binding to adult rat hepatocyte cell surfaces was done using cells isolated by two different collagenase perfusion methodologies and cultured as monolayers with two different tissue culture media formulations (protocol I vs . protocol II) . The presence of alpha-D-mannosyl and alpha-D-glucosyl groups was detected by the binding of Concanavalin A (Con A), Lens culinaris agglutinin (LCA), and Pisum sativum agglutinin (PSA) to freshly isolated cells . Furthermore, beta-D-galactose {Ricinus communis agglutinin (RCA)} and sialic acid residues {wheat germ (WGA)} were also found . Protocols I and II served as models for evaluation of: a) the stripping effect of collagenase separation procedures, b) the restoration in culture of collagenase-stripped sugar residues, c) the effect of the culture environment on cell viability {as measured by lactic acid dehydrogenase (LDH) leakage} and the protein content of hepatocytes, and d) the presence of cell surface sugar residues as a function of culture duration . The ultrastructural morphology of freshly isolated and cultured hepatocytes was also evaluated . These studies indicated that a decline in lectin binding invariably occurred earlier than a massive leakage of LDH and a decrease in the protein content of the cells in culture . Ultrastructurally, autophagocytosis was an early phenomenon in cells isolated and cultured by protocol I, which was also inferior to protocol II regarding the preservation of hepatocyte glycocalyces . Sugar residues lost due to the collagenase-stripping effect were restored, as shown by lectin binding, within the first 24 h of culture . This stripping effect was confirmed by quantitative evaluations of lectin binding to hepatocytes in culture after an incubation with collagenase . This study shows that the binding of peroxidase-labeled lectins is a useful tool for quantitative evaluation of the sugar composition of hepatocyte cultures. J Biol Chem, 1988 Apr 25, 263(12), 5846 - 52 Expression and dexamethasone regulation of the human corticotropin-releasing hormone gene in a mouse anterior pituitary cell line; Adler GK et al.; The factors controlling the expression of corticotropin-releasing hormone (CRH), a hypothalamic neuropeptide involved in the regulation of ACTH secretion, are poorly understood partly because a suitable in vitro model is lacking . To study the regulation of CRH gene expression, an 8-kilobase (kb) DNA fragment containing the entire human CRH gene as well as approximately 6 kb of 5' sequence and 0.8 kb of 3' sequence was isolated from a lambda Charon 4A human genomic library and introduced into a mouse anterior pituitary cell line, AtT-20, by CaPO4 transfection with a neomycin-selectable marker . Approximately 10% of the neomycin-resistant lines stably expressed the CRH gene and secreted radioimmunoassay-detectable CRH into culture media at levels greater than 100 pg/ml . By Southern blot analysis the 8-kb DNA fragment containing the CRH gene had been incorporated intact into the AtT-20 genome . In each CRH-producing strain, but not in the parent AtT-20 cell line, we detected by Northern blot analysis an RNA species that hybridized to two radioactive cRNA probes specific for either the 5' or 3' portion of CRH mRNA, and that co-migrated with placental CRH mRNA . Dexamethasone treatment for 24-96 h caused a specific decrease in CRH mRNA and peptide levels of 40-50% in the five CRH-producing cell lines with half-maximal suppression at approximately 10(-9) M dexamethasone, indicating that CRH gene expression is negatively regulated by glucocorticoids . Thus, we have established an in vitro model suitable for studying in detail those cis- and trans-acting factors which regulate CRH gene expression. Biochim Biophys Acta, 1988 Apr 15, 959(3), 220 - 8 Membrane-bound lipoprotein lipase on human monocyte-derived macrophages: localization by immunocolloidal gold technique; Goldberg IJ et al.; Macrophages from both rodent and human sources have been shown to produce lipoprotein lipase (LPL), the enzyme activity of which can be measured in culture media and in cellular homogenates . The studies reported here show the presence of LPL on the surface of human monocyte-derived macrophages . An inhibitory monoclonal antibody to human LPL was used for cellular and immunoelectron microscopy studies . This antibody is a competitive inhibitor of LPL hydrolysis of triacylglycerol but does not inhibit LPL hydrolysis of a water-soluble substrate, p-nitrophenyl acetate . Furthermore, when postheparin plasma was mixed with monoclonal antibody prior to gel filtration on 6% agarose, the LPL activity eluted with the lipoproteins and was not inhibited by the antibody . These studies suggest that the antibody recognized the lipid/lipoprotein binding site of the LPL molecule . Membrane-bound LPL was demonstrated on human monocyte-derived macrophages using colloidal gold-protein A to detect the monoclonal antibody to LPL . The surface colloidal gold was randomly distributed with a surface density of 56,700 gold particles per cell . Control cells cultured in heparin-containing media (10 units/ml) or cells reacted with anti-hepatic triacylglycerol lipase monoclonal IgG or nonimmune mouse IgG did not exhibit membrane binding of protein A-gold . Macrophages were incubated with control and monoclonal anti-LPL IgGs and 125I-labeled anti-mouse IgG F(ab')2 . Heparin-releasable membrane-bound anti-LPL antibody was demonstrated . These studies demonstrate the presence of LPL on the surface of human monocyte-derived macrophages, such that the LPL is oriented with its lipid-binding portion (recognized by this antibody) exposed . Membrane-associated LPL may be important in the interaction and subsequent uptake of lipid and lipoproteins by macrophages and in the generation of atherosclerotic foam cells. J In Vitro Fert Embryo Transf, 1988 Apr, 5(2), 76 - 80 Deterioration of stored culture media as monitored by a sperm motility bioassay; Stewart-Savage J et al.; A hamster sperm motility bioassay was used to monitor medium quality during storage . The media studied were (1) TL-PVA, a modified Tyrode's solution; (2) TLP-PVA, a defined medium that supports hamster sperm motility but does not support capacitation; and (3) TALP-PVA (TLP-PVA + serum albumin), which supports hamster sperm survival and capacitation . Each medium was stored at 5 and -20 degrees C and tested every two weeks . All of the media deteriorated with increased storage time, but at different rates (TLP-PVA greater than TL-PVA greater than TALP-PVA) . The deterioration of the media correlated with an increase in pH during storage, probably due to a loss of CO2 and bicarbonate . Albumin, added after storage, was able to rescue frozen TL-PVA. J In Vitro Fert Embryo Transf, 1988 Apr, 5(2), 67 - 75 A rapid sperm motility bioassay procedure for quality-control testing of water and culture media; Bavister BD et al.; A rapid bioassay procedure is described for quality-control testing water and apparatus used in the preparation of media for gamete and embryo culture . This bioassay is based on the sensitivity of hamster epididymal spermatozoa to contaminants present in water and/or in the culture apparatus . The bioassay is usually performed using a modified Tyrode's solution as the sperm culture medium, although complex media can be used . The sensitivity of this test is greatly enhanced by the absence of protein in the medium . The bioassay has been used to detect impurities in water prepared by a standard cartridge filtration system and to verify that reverse-osmosis pretreatment of water could eliminate the problem . It has also detected toxic contaminants leached from syringe filters during medium sterilization . The bioassay is simple to perform and can be completed in 1 working day . It may be a useful alternative to the conventional mouse embryo tests that are in widespread use in human in vitro fertilization (IVF) laboratories. J In Vitro Fert Embryo Transf, 1988 Apr, 5(2), 61 - 6 Influences of prolactin upon spermatogenesis and spermatozoa during in vitro fertilization in mice; Mori C et al.; Hyperprolactinemia, induced by pituitary isografts for 20 weeks in male mice and confirmed by radioimmunoassay using anti-mouse prolactin serum, did not impair spermatogenesis in the testis and maturing processes of spermatozoa in the epididymis . Incubation of freshly obtained epididymal spermatozoa for 90 min in culture media containing various levels of mouse prolactin did not yield any adverse effects on percentage motility rates of epididymal spermatozoa . When the level of mouse prolactin in the preincubation medium for epididymal spermatozoa was 100 ng/ml, the rate of fertilization by these preincubated spermatozoa in the subsequent in vitro fertilization experiment was significantly lowered compared with that observed in controls . However, when the level of prolactin in preincubation media was 10 ng/ml, no significant reduction in the rate of fertilization occurred . The present experiments seem to indicate the existence of some differences in the effects of prolactin on male germ cells until they reach the tail of the epididymis and on the processes of capacitation and/or fertilization by epididymal spermatozoa after they leave the epididymis. Mol Cell Biol, 1988 Apr, 8(4), 1845 - 8 Retinoic acid increases the sensitivity of the rat embryo fibroblast transformation assay; Halazonetis TD et al.; The rat embryo fibroblast focus assay is used to evaluate the transforming potential of several oncogenes . The sensitivity of this assay increased fivefold when retinoic acid was added to tissue culture media . Retinoic acid probably acts by selectively inhibiting the proliferation of nontransformed cells. Hum Reprod, 1988 Apr, 3(3), 367 - 76 Assessment of heterospecific zona-free ovum penetration under fully defined conditions; Tomkins PT et al.; The prognostic value of the sperm penetration assay (SPA) using zona-free hamster ova has not been universally accepted . This is partly due to technical problems with the assay, resulting in 5-17% false negative results . The advantage of replacing albumin with defined synthetic polymers in the SPA culture media, increasing the calcium gradient and incorporating potential active tract agents and antioxidants, has been investigated . These studies have resulted in the development of a procedure that has produced no negative results among known fertile donors (n = 55) . Application of this technique to a non-specific group of patients with suspected sub-fertility produced a different pattern of sperm penetration compared with the 'standard' SPA in 27% of cases . Improved sperm velocity, motile survival, enhanced hyaluronidase release and rate of acrosome induction were noted using the synthetic media. J Biomed Mater Res, 1988 Apr, 22(4), 271 - 85 Endothelialization of polymer surfaces; Absolom DR et al.; The role that substrate surface properties play in influencing the extent of endothelialization of polymer surfaces has been investigated . For a wide range of polymer surfaces, the degree of endothelialization for both porcine and bovine endothelial cells is directly related to polymer surface tension: increased endothelialization occurring with increasing substrate surface tension . As a result of adsorption of the proteins in the culture media, the surface properties of the polymers are altered considerably . The protein-coated polymers were characterized by means of liquid-liquid contact angle measurements under non-denaturing conditions . A striking correlation is observed between the degree of endothelialization and the measured dextran contact angle . The degree of endothelial cell spreading is not related to polymer surface tension . Cell morphology and extracellular matrix production, however, are influenced by substrate surface properties. Fertil Steril, 1988 Apr, 49(4), 676 - 9 High potassium concentration improves the rate of acrosome reaction in human spermatozoa; Roblero L et al.; Progressively motile spermatozoa recovered by swim-up method from semen of two fertile men were incubated for 24 hours in culture media containing either 4.7, 15, or 25 mM of potassium (K) . Aliquots of each culture condition were obtained at 0, 1, 5, 10, and 24 hours of incubation for the assessment of progressive motility, percentage of dead spermatozoa, and percentage of acrosome reaction (AR), as measured by triple-stain technique . A total of ten experiments including each K concentration were analyzed . The results of this study showed no effect of K concentration on the percentage of progressively motile spermatozoa, irrespective of the time of incubation . The percentage of live spermatozoa was significantly greater in culture medium containing 25 mM K (P less than 0.05) . There was a greater percentage of reacted spermatozoa with 25 mM K, as compared with 4.7 and 15 mM (P less than 0.05) . Furthermore, the time taken to achieve 20% of AR was 2 hours at 25 mM K compared with 10.9 hours at 4.7 mM K. Cell Biochem Funct, 1988 Apr, 6(2), 101 - 5 Effects of insulin and dexamethasone on adenine nucleotide levels in cultured hepatocytes from adult rat; Gallo G et al.; Insulin and dexamethasone, usually added to culture media, play an important role in maintaining the survival of functional hepatocytes . Adenine nucleotide concentrations and energy charge values of cultured hepatocytes were determined to investigate the relationship between the beneficial effects of these hormones and the energy status of the cells . The results indicate that insulin and dexamethasone are essential in maintaining the metabolic competence of cultured hepatocytes and that this correlates with the absolute concentration of ATP rather than with the energy charge. Mech Ageing Dev, 1988 Apr, 43(1), 25 - 44 The aging of the endocrine pancreas of the rat . II . Cytoplasmic parameters of the B-cell, including insulin synthesis and secretion; de Clercq L et al.; Comparative ultrastructural stereology of 6 and 24-month-old rat B-cell cytoplasm revealed an increase with age in secondary lysosomes and a decrease in the volume density of RER and Golgi apparatus . The reduction of RER observed in freshly isolated islets could affect (pro)insulin biosynthesis in vitro: if the initial mobilization of precursor molecules for protein synthesis was the same, a delay was noted in their transit to the Golgi apparatus in B-cells of old islets . No further differences were seen in the autoradiographic distribution of radioactive amino-acids . More, the stock of insulin granules was similar in all age groups in both in vivo and in vitro conditions . Neither were any differences observed in the insulin secretion into culture media as well as during a subsequent incubation in supraphysiological glucose concentrations. In Vitro Cell Dev Biol, 1988 Apr, 24(4), 299 - 303 Growth of H-35 rat hepatoma cells in unsupplemented serum-free media: effect of transferrin, insulin and cell density; Shapiro LE et al.; Serum-free tissue culture medium consisting of a 1:1 mixture of Dulbecco's modified Eagle's medium (DMEM) and Ham's F12 medium is herein shown to support growth of Reuber H-35 cells over several days in culture . Cells were initially plated in serum containing DMEM medium for 3 h . After cell attachment, serum is removed and replaced with a serum-free 1:1 mixture of these two commercially available tissue culture media . The doubling time of cell growth in this unsupplemented serum-free medium was 46 h in lightly plated cultures over the first 5 d . The presence of transferrin (5 micrograms/ml) and insulin (3.3 nM) results in a cell doubling time of 17 h, which equaled the growth rate in medium containing 10% fetal bovine serum . In the absence of transferrin, growth rates in serum-free medium were correlated with the cell density of cultures . Conditioned medium from dense, serum-free cultures has growth-stimulating activity in recipient lightly plated cultures . This simple, serum-free culture medium will facilitate studies on the growth regulation of H-35 rat hepatoma cells. Endocrinology, 1988 Apr, 122(4), 1639 - 45 The in vitro synthesis and release of proteins by the human oviduct; Verhage HG et al.; The purpose of this study was to identify major proteins synthesized by the human oviduct and to determine if the ability of the human oviduct to synthesize these proteins was correlated with a specific stage(s) of the menstrual cycle . Oviducts, obtained from normal menstrual and immediately postpartum women, were minced and cultured in the presence of labeled precursors . The culture medium was analyzed for newly synthesized proteins by electrophoresis, followed by fluorography . One-dimensional gel electrophoretic analysis of culture medium of oviducts obtained at midcycle revealed a major diffuse band in the 120,000-130,000 mol wt (Mr) region which was greatly diminished in intensity or nondetectable in the media of oviducts obtained at all other stages of the cycle . Two-dimensional gel electrophoresis resolved the 120,000-130,000 Mr region into two major proteins, one basic and one acidic . Both proteins were intensely labeled with glucosamine, and the basic protein also incorporated leucine and methionine . Western blots of culture media incubated with antibodies against human placental proteins (PP) demonstrated that PP4 and PP7 were released throughout the cycle, while PP14 was present only at the late luteal stage of the cycle . This study demonstrates that human oviducts continue to synthesize and release macromolecules during organ culture . Additionally, the synthesis of some of these proteins appeared to be stage specific . The Mr 120,000-130,000 glycoproteins are of particular interest since they were observed in medium from midcycle oviducts, a period when the oviduct participates in gamete transport and embryo development. Endocrinology, 1988 Apr, 122(4), 1603 - 12 Somatomedin-C/insulin-like growth factor I as an enhancer of androgen biosynthesis by cultured rat ovarian cells; Hernandez ER et al.; The ovarian granulosa cell has recently been shown to be a site of somatomedin-C/insulin-like growth factor I (Sm-C/IGF-I) production, reception, and action . These observations have generally been interpreted to suggest the existence of an autocrine loop concerned with granulosa cell physiology . It is the objective of the in vitro studies reported herein to extend these observations by evaluating the interaction of Sm-C/IGF-I with the adjacent thecal-interstitial cell . Treatment of collagenase-processed whole ovarian dispersates or highly enriched (greater than 90%) thecal-interstitial cells from immature rats with Sm-C/IGF-I (50 ng/ml) or hCG (1 ng/ml), resulted in 2.1- and 4.0-fold increments in the accumulation of androsterone (3 alpha-hydroxy-5 alpha-androstane-17-one), the main androgenic steroid identified in culture media . However, combined treatment with both agents unmasked a synergistic interaction producing a 3.3-fold increase in the hCG-stimulated accumulation of androsterone, an effect consequent to enhanced androgen biosynthesis rather than diminished degradation . Unaccounted for by an increase in viable ovarian cell numbers and independent of the hCG dose (0.1-10 ng/ml) used, the Sm-C/IGF-I effect proved time and dose dependent, with a projected minimal effective dose of 3 ng/ml and a minimal time requirement of 72 h . {125I}Iodo-Sm-C/IGF-I binding to untreated highly enriched thecal-interstitial cells proved saturable, with a single class (Hill coefficient = 0.98 +/- 0.01) of high affinity (Kd = 3.0 nM), low capacity (maximum binding = 10,840 +/- 2,108 sites/cell) binding sites . Limited specificity studies using related peptides produced a rank order of competitive potency of: Sm-C/IGF-I greater than multiplication stimulating activity greater than insulin, a pattern compatible with the presence of type I IGF receptors . Other related peptides, such as porcine proinsulin and porcine desoctapeptide insulin, proved weakly effective in inhibiting Sm-C/IGF-I binding to its receptor; unrelated peptides such as porcine relaxin and erythropoietin were without effect . Taken together, these findings suggest that 1) the thecal-interstitial cell, like the granulosa cell, may be a site of Sm-C/IGF-I reception and action, and 2) the ability of high dose insulin to stimulate ovarian androgen biosynthesis may be due to its capacity to act as a Sm-C/IGF-I surrogate, its high dose requirements reflecting cross-interaction with the type I receptor.(ABSTRACT TRUNCATED AT 400 WORDS) J Immunol, 1988 Apr 1, 140(7), 2274 - 8 The expression and modulation of IL-1 alpha in murine keratinocytes; Ansel JC et al.; Murine and human keratinocytes produce an IL-1-like factor that appears to be similar if not identical to monocyte-derived IL-1 . IL-1 may be an important mediator in cutaneous inflammatory responses, however, little is currently known concerning factors that may modulate IL-1 expression in keratinocytes . To address this issue we examined the effect of LPS, UV, and the cell differentiation state on murine keratinocyte IL-1 mRNA expression . Our results indicated that as with the murine P388D1 monocyte cell line, PAM 212 keratinocytes constitutively express abundant amounts of IL-1 alpha mRNA . On exposure to LPS (100 micrograms/ml) for 8 h there was more than 10 times the increase in PAM 212 IL-1 alpha mRNA which was accompanied by a sixfold increase in supernatant IL-1 activity . Similarly UV irradiation had a significant effect on keratinocyte IL-1 alpha expression . High dose UV (300 mJ/cm2) inhibited PAM 212 IL-1 alpha expression at 4, 8, 24, 48 h post-UV whereas a lower dose of UV (100 mJ/cm2) inhibited UV at 4 and 8 h post-UV, but induced IL-1 expression at 24 and 48 h post-UV . The expression of IL-1 alpha varied with the differentiation state of the keratinocytes . Freshly removed newborn murine keratinocytes were found to constitutively express IL-1 alpha mRNA . Keratinocytes grown in low {Ca2+} tissue culture media (0.05 mM) for 6 days, functionally and phenotypically become undifferentiated and express increased quantities of IL-1 alpha mRNA, whereas cells grown in high {Ca2+} media (1.2 mM) for 6 days become terminally differentiated and IL-1 expression ceased . Keratinocytes cultured for 3 days in low {Ca2+} conditions expressed an intermediate level of IL-1 alpha . In contrast, little or no IL-1 beta mRNA was detected in either the PAM 212 cells or newborn murine keratinocytes . Thus LPS, UV, and cell differentiation state have a significant effect on expression of IL-1 alpha in murine keratinocytes. Prostaglandins, 1988 Apr, 35(4), 535 - 48 Role of phospholipase C in mediating oxytocin-induced release of prostaglandin F2 alpha from ovine endometrial tissue; Silvia WJ et al.; Two experiments were conducted to determine if the ability of oxytocin to stimulate release of prostaglandin (PG)F2 alpha from ovine uterine tissue involved activation of phospholipase C (PLC) . In the first experiment, 9 ewes were injected with progesterone for 11 d (12 mg/d, im) . On days 11 and 12, ewes received an injection of estradiol (100 micrograms, im) . Caruncular endometrial tissue was collected on d 13 and incubated in the presence or absence of oxytocin (10(-6) M) . Concentrations of PGF2 alpha and its metabolite, 13,14-dihydro-15-keto-PGF2 alpha (PGFM), in culture media were determined by radioimmunoassay . PLC activity was determined by measuring the intracellular accumulation of 3H-inositol phosphates after preincubation with 3H-inositol . Concentrations of PGF2 alpha and total PGF (PGF2 alpha + PGFM) in culture media were greater for explants treated with oxytocin than for controls (p . less than .02, p less than .06, respectively) . A similar effect of oxytocin on intracellular concentrations of 3H-inositol phosphates was observed (p less than .01) . A second experiment was conducted to determine if agonists of second messengers, produced by activation of PLC, could stimulate release of PGF2 alpha from ovine endometrial tissue . Seven ewes were treated with progesterone and estradiol as in experiment 1 . Explants of caruncular tissue from each ewe were incubated with 1) control medium, 2) A23187 (10(-5) M), 3) oxytocin (10(-6) M), 4) phorbol 12-myristate 13-acetate (PMA, 10(-7) M), 5) PMA + A23187 and 6) PMA + oxytocin . Significant stimulatory effects of oxytocin, PMA and A23187 on concentrations of PGF2 alpha and total PGF in culture media were observed (p . less than .05, p less than .1, p less than .1, respectively) . In conclusion, oxytocin stimulated release of PGF2 alpha and activity of PLC in explants of ovine endometrial tissue in vitro . Second messengers associated with activation of PLC enhanced release of PGF2 alpha from ovine endometrial tissue. Blood, 1988 Apr, 71(4), 899 - 906 Enhanced expression of transforming growth factor beta during megakaryoblastic differentiation of K562 leukemia cells; Alitalo R et al.; Platelet alpha granules contain several growth factors such as the transforming growth factor beta (TGF-beta) that are released during blood clotting and are thought to participate in the repair of tissue injury; however, the site of synthesis of platelet TGF-beta has not been demonstrated . We studied TGF-beta expression during megakaryoblastic differentiation of the chronic myeloid leukemia cell line K562 in vitro . These cells have mainly erythroid characteristics but acquire several megakaryoblastic properties when treated with the phorbol diester 12-0-tetradecanoyl-13-phorbolacetate (TPA) . During four subsequent days of megakaryoblastic differentiation the amount of the 2.5-kilobase (kb) TGF-beta mRNA increased about eightfold, and a novel 2.3-kb mRNA species was induced in the K562 cells . This occurred concomitantly with distinct induction patterns of platelet-derived growth factor A (PDGF-A) and c-sis (PDGF-B chain) RNAs and several platelet antigens . The expression of erythroid markers such as glycophorin A decreased . Culture media of TPA-differentiated K562 cells also contained TGF-beta polypeptides as shown by a sensitive radioreceptor assay and by immunoprecipitation after metabolic labeling of the cells . These polypeptides were not seen in culture media from dimethyl sulfoxide- or sodium butyrate-treated cells . Unlike in several other cells, exogenously added TGF-beta 1 or 2 affected neither TGF-beta nor PDGF RNA expression in K562 cells. Eur J Pediatr, 1988 Apr, 147(3), 321 - 7 A variant of mucolipidosis . II . Clinical, biochemical and pathological investigations; Poenaru L et al.; We present in this paper a patient with a clinically intermediate form of mucolipidosis (ML) . Lysosomal hydrolase activity in fibroblasts was normal and levels of these enzymes in culture media were not elevated . There was a striking elevation of several hydrolases in serum and a deficiency (15% of normal) of N-acetyl-glucosamine phosphotransferase in fibroblasts . Atypical electron microscopic findings were also observed . There was no evidence of increased synthesis, slower turnover, unbalanced distribution or further changes in lysosomal enzymes . Phosphotransferase deficiency against endogenous beta-glucosaminidase and the fact that the electrophoretic mobility of lysosomal enzymes was identical to that of MLII suggest that these enzymes are not phosphorylated . Hypotheses that could explain this atypical pathology are discussed. J Biol Chem, 1988 Mar 25, 263(9), 4259 - 62 Affinity-based chromatography utilizing genetically engineered proteins . Interaction of Bordetella pertussis adenylate cyclase with calmodulin; Haiech J et al.; An engineered calmodulin differs from vertebrate calmodulin in its ability to activate Bordetella pertussis adenylate cyclase, and this difference has been utilized as the basis for a new purification protocol for the adenylate cyclase . VU-8 calmodulin, in which 3 glutamic acid residues (residues 82-84) have been substituted with 3 lysine residues, has a 1000-fold lower apparent affinity for the adenylate cyclase, compared to vertebrate calmodulin, and decreased maximal activity . Because of the relatively calcium-independent nature of the interaction between calmodulin and the cyclase, the use of calmodulin-Sepharose conjugates in the purification of the cyclase requires the use of chaotropic agents for elution . However, when immobilized VU-8 calmodulin was tested as a calcium-dependent, affinity-based, adsorption chromatography step in the purification of the cyclase from culture media or bacterial extracts, the enzyme bound to the column in a calcium-dependent manner, and a nearly homogeneous enzyme was obtained in high yield . These results demonstrate the feasibility of using engineered calmodulins that have selective differences in activity for the rational design of rapid purification protocols for calmodulin-binding proteins as well as indicate the importance of the conserved negative charge cluster at residues 82-84 of calmodulin for activation of this cyclase. J Immunol Methods, 1988 Mar 16, 107(2), 157 - 63 Modification and application of a simple, surface hydrophobicity detection method to immune cells; Hazen BW et al.; A simple method to assess fungal cell surface hydrophobicity involves enumeration of cell-attached, polystyrene latex microspheres . Modifications and conditions of this method for application to immune cell populations were investigated . The media used for suspending cells and microspheres appeared to influence microsphere attachment . Several tissue culture media supported high levels of microsphere attachment, but serum inhibited attachment . The concentration of microspheres also influenced the apparent level of detectable cell surface hydrophobicity . Under conditions which allow different levels of apparent cell surface hydrophobicity to be discriminated, the assay revealed that surface hydrophobicity of YAC-1 cells, which are used as standard targets in murine natural killer cell assays, varied depending on the time of harvest during growth and that phagocytic populations differed in cell surface hydrophobicity . Trypsinization experiments indicated that one hydrophobic constituent of the cell surface includes protein . These results indicate that the microsphere assay is a useful method for assessing cell surface hydrophobicity . The possibility that the assay could be used to determine quantitatively the surface free energies of immune cells and cell targets is discussed. Cancer Res, 1988 Mar 15, 48(6), 1505 - 11 Purification and composition of a novel gastrointestinal tumor-associated glycoprotein expressing sialylated lacto-N-fucopentaose II (CA 19-9); Klug TL et al.; Monoclonal antibody 1116NS 19-9 (Mab 19-9) exhibits selective reactivity with human gastrointestinal carcinomas and recognizes a carbohydrate determinant (CA 19-9) defined as a sialylated lacto-N-fucopentaose II . A scheme was devised for the purification of a human gastrointestinal tumor-associated glycoprotein antigen expressing CA 19-9 from colorectal carcinoma cell line SW1116 culture media in high yield . The key steps in the purification were immunoaffinity column chromatography with Mab 19-9 followed by reduction and alkylation of the specifically bound proteins in the presence of 6 M guanidine hydrochloride and a second Mab 19-9 immunoaffinity fractionation . The purified CA 19-9 containing glycoprotein ran as a single band on sodium dodecyl sulfate-polyacrylamide gradient gels with an apparent molecular mass of 210 kilodaltons . In the absence of detergents, this purified glycoprotein apparently reassociated to form aggregates of 600-2000 kilodaltons molecular mass as determined by size-exclusion chromatography . Amino acid analysis of CA 19-9 containing glycoprotein revealed that serine, threonine, and proline together accounted for greater than 35% of the amino acid residues, consistent with a mucin-like structure for the protein . Carbohydrate compositional analysis, however, was in contrast to a typical mucin with a fucose:mannose:galactose:N-acetylgalactosamine: N-acetylglucosamine:N-acetylneuraminic acid molar ratio of 4:1:12:2.5:5:5 . The presence of both N-acetylgalactosamine and mannose suggested that both O- and N-linked oligosaccharides may exist on CA 19-9 containing glycoprotein . Protein and carbohydrate analyses indicated that this novel tumor-associated glycoprotein was 85% carbohydrate by weight . This purification procedure may be applicable to the isolation of other epithelial tumor-associated antigens. Andrologia, 1988 Mar-Apr, 20(2), 169 - 72 ATP-content and kinetics of acrosome reaction in human spermatozoa: influence of various culture media and incubation time; Dolci S et al.; The ATP content and the kinetics of acrosome reaction of isolated human spermatozoa were investigated during an in vitro culture period of 3 hours in 3 serum-enriched media commonly adopted in IVF and GIFT procedures (Earle solution, Ham F10 and Menezo B2) . The ATP concentration in spermatozoa did not change between 1 and 3 hr of incubation and no differences were seen using the three media under investigation . The percentage of acrosome reactions significantly increased during culture, to a similar extent in the three media . The results suggest that each of the three media is equally well suitable for human sperm culture in preparation of IVF and GIFT procedures. Immunol Lett, 1988 Mar, 17(3), 217 - 22 Sulfhydryl groups generated by macrophages into the culture medium; Jokay I et al.; Previous data showing that thiols can functionally replace macrophages in certain in vitro lymphocyte reactions have raised the possibility that macrophages generate SH groups in the medium . The SH activity of cell-free medium decreases considerably due to auto-oxidation . The presence of macrophages inhibited the spontaneous decrease of SH activity and even increased the number of free SH groups in the culture media . This was designated as SH generation . Resident as well as in vivo stimulated (NaIO4, thioglycollate medium, paraffin oil or BCG) macrophages have nearly the same capacity to generate SH groups when cultured in vitro . The SH-generating ability of macrophages depends on the viability and density of cells, and is greatly influenced by the serum concentration of the media . The majority of SH groups produced by macrophages could be demonstrated as albumin SH groups and proved to be more resistant to autooxidation than non-protein thiols . Moreover albumin could replace whole serum in respect of SH generation . It is suggested that macrophages by generating SH activity, may exert a non-specific helper function to lymphocytes or other cells in their environment. Mech Ageing Dev, 1988 Mar, 42(3), 215 - 27 Culture media variation as related to in vitro aging of human fibroblasts: II . Effects on nucleolar number/cell, volume/nucleolus and total nucleolar volume/cell; Weinstein ME et al.; The relative effect of five commonly used culture media (MEM, BME, McCoy's 5A, M199 and HMEM) on the average nucleolar number/cell, the average volume/nucleolus and the total nucleolar volume/cell was examined during in vitro senescence of WI-38 human fetal fibroblasts . Statistical analyses show that cells aging in MEM show a higher number of nucleoli/cell than that of cells aging in any other medium . For cells aging in the other four media, there are no significant differences in the average number of nucleoli/cell . Linear regression analysis shows that in all cases there is a linear decrease in the average number of nucleoli/cell as a function of PDL . Statistical analyses show that the average volume/nucleolus is significantly greater for cells aging in M199 than in any other medium . Cells aging in HMEM show smaller average nucleolar volume than cells aging in M199, but display larger volumes than that of cells aged in BME, McCoy's 5A, or MEM . Cells aging in BME and McCoy's 5A media show no significant difference among each other in terms of average nucleolar volume, but a difference in this parameter is noted in cells aging in BME and MEM . A linear regression analysis shows that the average volume/nucleolus increases linearly as a function of age for cells grown in all five media . Analysis of the total nucleolar volume/cell in the five media shows that cells aging in M199 and HMEM are not significantly different from each other in terms of this variable, but show significantly larger volumes than those of cells aging in BME, McCoy's 5A and MEM . Cells aging in BME, McCoy's 5A and MEM display no significant difference with regard to this parameter . Linear regression analysis shows a positive linear relationship between the PDL and the total nucleolar volume/cell . The relative effects of all five media are not the same on the three cellular variables studied during in vitro aging of WI-38 cells . We, therefore, suggest that one should note this medium differential in order to allow meaningful comparison of results on possible changes in various morphological parameters during in vitro senescence of diploid human fibroblasts. J Cell Biol, 1988 Mar, 106(3), 883 - 91 Mitogen activation induces the enhanced synthesis of two heat-shock proteins in human lymphocytes; Haire RN et al.; We have used mitogenic lectin (PHA) and a monoclonal antibody (OKT3) to stimulate human peripheral blood (G0) lymphocytes, in the presence of monocytes, and have found two major preferentially synthesized proteins, 73 and 95 kD, which are induced by the mitogens . The elevated synthesis of both proteins begins approximately 4-6 h after mitogen addition (early to mid G0/G1) before entry into first S phase . Maximum synthesis of both proteins is reached by 12 h after mitogen addition when P95 synthesis represents approximately 4%, and P73 approximately 2%, of the total protein synthesis, compared with less than 0.5% for each protein in cells cultured without mitogen . Thus, the proteins appear to be major components of activated cells . We find that both P73 and P95 are induced by heat stress as well as mitogenic stimulation . The induction of the proteins is not affected by either deleting glucose from the culture media or, alternatively, by supplementing it . Using polyclonal antibodies prepared to each of the proteins isolated from mitogen activated cells and monoclonal antibodies that were raised to heat shock proteins, we are able to show that P95 is electrophoretically and immunologically identical to the HSP 90 induced by heat stress . P73 is one of the 70 kD HSPs, (termed HSC 70; Pelham, H . R . B . 1986 . Cell . 46: 959-961), but is different from the most strongly heat inducible form of HSP 70 (72 kD) . The distribution of both proteins in subcellular fractions of mitogen activated lymphocytes is similar to the reported localization of the respective HSP's in other cell types . The results suggest that HSP 90 and HSC 70 may have functional roles in stress response and growth processes of human lymphocytes. Andrologia, 1988 Mar-Apr, 20(2), 173 - 81 Paracrine factors in adult rat testis gonadotrophin control of opioids and LHRH like peptide; Saint Pol P et al.; A paracrine regulation involves agents which are produced by one cell type and act on an other one within an organ . In rodent testis, local control mechanisms modulate the actions of the gonadotrophins according to local requirements . Two groups of peptides-opioids and testicular LHRH are defined as paracrine factors and in vivo they are both modified by HCG . In vitro, after HCG exposure, we first localized an opioid like material in Sertoli cells cytoplasma by immunohistochemistry . This material is detected in freeze dried homologous culture media using a dot immunobinding technique . With a longer HCG exposure, an LHRH like material is then visualized in the basal compartment of the Sertoli cells and it is detected in freeze dried homologous culture media by the same technical procedure than for opioid material . By adding synthetic enkephalins to culture medium, we obtain the same results as with the endogenous opioid material, excreted after HCG addition . If naloxone a potent opiate antagonist, is added to the culture medium previously to HCG or enkephalins, the Sertoli cells cytoplasma are no more immunoreactives with the anti-enkephalin serum and no LHRH material is neither visualized by immunohistochemical technique neither detected in culture media . We conclude that testicular opioids, synthetized by the Leydig cells and which have specific Sertoli cells receptors are one Leydig-Sertoli paracrine communication factor . One way of response to their receptor fixation is the synthesis and excretion by Sertoli cells of testicular LHRH . This one is known to act on Leydig cells via specific receptors and it is one Sertoli-Leydig cells paracrine communication factor.(ABSTRACT TRUNCATED AT 250 WORDS) Mol Cell Endocrinol, 1988 Mar, 56(1-2), 35 - 40 Selective control of rat granulosa cell inhibin production by FSH and LH in vitro; Zhang Z et al.; We studied the role of luteinising hormone (LH) and human chorionic gonadotropin (hCG) in regulation of rat granulosa cell inhibin production . Whereas pregnant mare serum gonadotropin (PMSG) or purified rat follicle-stimulating hormone (FSH) stimulated inhibin accumulation in culture media up to 6-fold in a dose-dependent manner, hCG or LH alone were without significant effect . Concomitant addition of hCG or LH to cultures containing half maximal or maximal stimulating concentrations of PMSG did, however, result in a dose-dependent inhibition of PMSG/FSH-induced inhibin production . However, in the 2-step culture system a biphasic effect of hCG treatment on FSH-primed granulosa cell inhibin and progesterone production was evident . hCG was only inhibitory when the cells in the 2-step system were primed with PMSG . These data indicate that granulosa cell inhibin production is under direct control of both FSH and LH, and provide a possible explanation for alterations in inhibin activity around the time of ovulation in vivo. Am J Pathol, 1988 Mar, 130(3), 427 - 30 Tumor necrosis factor enhances interferon-induced Ia antigen expression on murine islet parenchymal cells; Wright JR Jr et al.; Islets from male B10.BR mice (H-2k) were isolated by the collagenase technique, hand-picked with a Pasteur pipette, and incubated in tissue culture media supplemented with recombinant murine interferon-gamma (rIFN-gamma), recombinant murine tumor necrosis factor (rTNF), or both . IAk-molecules could be identified on the surface of islets incubated for 5 days with a combination of rIFN-gamma (1, 10, or 100 ng/ml) and rTNF (10 or 50 U/ml) by indirect immunofluorescence . Optimal concentrations of rIFN-gamma and rTNF, 10 ng/ml and 50 U/ml, respectively, were used in all subsequent experiments . Weak Ia-positivity could be identified on the surface of islets cultured with both cytokines for as little as 48 hours; however, the staining appeared most intense after 5 days of culture . Intensely Ia-positive islets were then carefully washed and cultured in media without either cytokine; Ia positivity could be identified on the surface of these islets for up to 1 week . Dispersed islet cells were cultured with rIFN-gamma alone (10 ng/ml), rTNF alone (50 U/ml), or both cytokines for 5 or 10 days . After either 5 or 10 days of culture with both cytokines, intense immunofluorescent staining for Ia could be identified on the surface of greater than 80-90% of the viable islet cells . Culture with IFN alone for 10 days resulted in 15-20% Ia positivity; culture with TNF alone did not cause Ia expression. Fertil Steril, 1988 Mar, 49(3), 516 - 21 Mouse embryo culture as quality control for human in vitro fertilization: the one-cell versus the two-cell model; Davidson A et al.; Female mice were superovulated with pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG) and mated with male mice . One-cell (n = 429) and 2-cell (n = 450) embryos were collected 20 and 42 hours after hCG and cultured in Ham's F-10 medium (Gibco, Grand Island, NY) (282 mOsm/l, pH 7.4) and in media of altered osmolality (260, 300, 316 mOsm/l), altered pH (7.0, 7.8, 8.0) or various dilutions of Cidex (Surgikos, Arlington, TX) (1:1000, 1:10,000, 1:100,000) . Stages of development were observed for 4 days . The development of embryos in the 1-cell system was significantly impaired under all studied conditions by the 4-cell stage of development . The 2-cell system failed to detect trace amounts of Cidex in the culture media and an increase in osmolality to 300 mOsm/l . Other changes in osmolality (260 mOsm/l) and pH (7.8) were detected by the 2-cell system only at the blastocyst stage . The authors conclude that the 1-cell system is more sensitive than the 2-cell system to mild changes in in vitro fertilization culture media. J Neuroimmunol, 1988 Mar, 17(4), 323 - 30 Activated suppressor cell function in multiple sclerosis--clinical correlations; Antel J et al.; Activated suppressor cell function mediated by either freshly isolated peripheral blood mononuclear cells (MNCs), freshly isolated CD8+ lymphocytes or by CD8+ cell lines, has previously been found to be reduced compared to controls in multiple sclerosis (MS) patients with progressive disease (MS-P) . In this study, we found that suppressor activity mediated by CD8+ cell lines, derived from MS patients with stable disease (MS-S) patients and maintained in culture for 14 days, was significantly greater (45 +/- 6%) compared to that mediated by MS-P patients' CD8+ cells (11 +/- 4%, P less than 0.005) . The MS-S suppressor values were, however, suggestively reduced compared to controls (60 +/- 6%, P less than 0.05) . MNC-mediated suppressor values for the MS-S group (61 +/- 5%) did not differ from the control group (67 +/- 6%) . Values for the MS-P group (7 +/- 6%) were significantly reduced compared to MS-S and control groups . Cytotoxic activity mediated by CD8+ cell lines showing defective suppressor function did not differ from control values . The cell lines in MS and control did not differ with respect to their rate of proliferation in the presence of IL-2 and OKT3 . Suppressor function in this assay was ablated if exogenous IL-2 was removed from the culture media . These data suggest that defective activated suppressor function is characteristic of the progressive form of MS, although a suppressor defect is also partially expressed in stable MS patients when CD8+ cell lines are studied. Acta Trop, 1988 Mar, 45(1), 67 - 76 Moulting and exsheathment of the infective larvae of Onchocerca lienalis (Filarioidea) in vitro; Pudney M et al.; Fourth-stage larvae (L4) of the cattle filarial parasite Onchocerca lienalis were produced from third stage forms (L3) and maintained in vitro using a variety of culture media and feeder cell layers . Up to 50% of the L3 larvae moulted to the fourth stage in the presence of two jird (Meriones unguiculatus) cell lines, a monkey kidney cell line and also with Vero cells . Moulting took place 2-5 days after of the initiation of the cultures: These L4 parasites could be maintained in a healthy condition for up to 88 days, although no significant increases in size of these larvae were observed during the culture period . The medium giving the most promising results, both in terms of moulting and survival, was 199 supplemented with heat inactivated foetal bovine serum (20%) and glucose (2 mg/ml) . The feeder layer cells could be replaced by fresh jird red blood cells . Moulting of parasites occurred in medium alone but the viability was reduced under these conditions. Am J Pathol, 1988 Mar, 130(3), 489 - 95 Macrophage-derived cytokines amplify immune complex-triggered O2- . responses by rat alveolar macrophages; Warren JS et al.; Interleukin-1 (IL-1) and tumor necrosis factor (TNF) are monocyte macrophage-derived hormonelike regulatory proteins that participate in many physiologic and pathophysiologic processes . Several proinflammatory activities have been attributed to these cytokines, but their importance in anatomically compartmentalized inflammatory processes is unclear . The current in vitro studies have been designed to examine modulatory influences of these cytokines on O2- . responses of rat phagocytes implicated as effector cells in immune complex mediated lung injury . Purified human IL-1, recombinant human TNF (rTNF), and culture supernatant from zymosan-activated alveolar macrophages significantly amplified O2- . responses of immune complex-stimulated alveolar macrophages but did not enhance the responses of neutrophils . Equivalent concentrations of IL-1, rTNF, and alveolar macrophage culture supernatant had no direct stimulatory effect on alveolar macrophages as measured by O2- . production . Culture media from unstimulated alveolar macrophages exerted negligible effects on O2- . generation by immune complex-activated alveolar macrophages . These data indicate that O2- responses of immune complex alveolar macrophages can be enhanced by the presence of IL-1, TNF, or media from activated macrophages . It is possible that macrophage products may greatly amplify tissue injury through the enhancement of oxygen radical production. J Chromatogr, 1988 Feb 26, 424(2), 315 - 23 Simple isocratic high-performance liquid chromatographic method for the separation of the six vitamers of vitamin B6; Argoudelis CJ; A simple, sensitive and fast isocratic high-performance liquid chromatographic method has been developed for the separation of all six biologically active forms (vitamers) of vitamin B6 . The separation is accomplished using a strong cation-exchange column and a mobile phase of 0.1 M ammonium dihydrogenphosphate adjusted to pH 4.0 . All six vitamers are separated within 20 min at a flowrate of 1 ml/min . The concentration of the vitamers is determined with a fluorescence detector (excitation 290 nm; emission 389 nm) . The within-run precision of the method expressed as the coefficient of variation is below 5% at the 25 pmol level . Pyridoxal 5'-phosphate can be determined using either pre- or post-column derivatization with sodium bisulfite . Application of the method to cell-free yeast culture media is presented. JAMA, 1988 Feb 26, 259(8), 1223 - 7 Diagnosis of trichomoniasis . Comparison of conventional wet-mount examination with cytologic studies, cultures, and monoclonal antibody staining of direct specimens; Krieger JN et al.; The accuracy of (1) conventional wet-mount examination, (2) Papanicolaou-stained gynecologic smears, (3) a direct slide test using fluorescein-conjugated monoclonal antibodies against Trichomonas vaginalis, and (4) two different culture media for the diagnosis of trichomoniasis in a high-risk population of 600 women was compared . Use of Feinberg-Whittington or Diamond's culture medium resulted in a diagnosis of 82 and 78 cases, respectively, and the combination of two cultures identified 88 infected women . In comparison, wet-mount examination detected only 53 (60%) of the cases . Cytologic smears were interpreted as positive for T vaginalis in 49 (56%) of the 88 cases but also resulted in seven false-positive smears, and specimens from 18 women with negative cultures were interpreted as "suspicious" for trichomoniasis . Monoclonal antibody staining detected 76 (86%) of the 88 positive specimens, including 27 (77%) of the 35 cases missed by wet-mount examination . In summary, wet-mount and cytologic studies were insensitive, and cytology study was the least specific method for diagnosis of trichomoniasis . Direct immunofluorescence with monoclonal antibodies holds promise as a sensitive and specific alternative to cultures for rapid detection of T vaginalis in clinical specimens. Cancer Res, 1988 Feb 15, 48(4), 822 - 5 Role of oxygen radicals in 12-O-tetradecanoylphorbol-13-acetate-induced squamous differentiation of cultured normal human bronchial epithelial cells; Gabrielson EW et al.; The tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) inhibits growth and induces terminal squamous differentiation of normal human bronchial cells when added to the culture media {J . C . Willey, A . J . Saladino, C . Ozanne, J . F . Lechner, and C . C . Harris, Carcinogenesis (lond.), 5: 209-215, 1984} . We have investigated the possibility of oxygen free radicals being involved as intermediates in this process . Electron paramagnetic resonance measurements using the spin-trapping agent 5,5-dimethyl-1-pyrroline-1-oxide failed to detect oxygen free radicals in bronchial epithelial cells exposed to TPA, although oxy radicals were detected in bronchial epithelial cells after a nontoxic exposure to menadione, and in human neutrophils after exposure to TPA . Addition to the culture media of free radical scavenger, i.e., reduced glutathione, N-acetylcysteine, D-alpha-tocopherol, copper (II) (3,5-diisopropylsalicyclic acid)2, or the combination of superoxide dismutase and catalase did not affect the dose-dependent growth inhibition of TPA on the bronchial epithelial cells . Moreover, exposure of the bronchial epithelial cells to TPA did not result in increased DNA single strand breaks measured by alkaline elution, as would be expected with a free radical mediated mechanism . Thus, our results argue against the importance of oxygen free radicals in the inhibition of growth and the induction of squamous differentiation by TPA in normal human bronchial epithelial cells. Hybridoma, 1988 Feb, 7(1), 69 - 77 Effects of culture media on murine hybridomas: definition of optimal conditions for hybridoma viability, cellular proliferation, and antibody production; Long WJ et al.; Different cell culture media were compared for their ability to support and promote the growth of stable hybridoma cell lines derived from three commonly used parental murine myelomas . Supplemented Dulbecco's modified Eagle's media (DMEM) and RPMI 1640 media were studied . The DMEM-based media were found to support greater numbers of cells for longer time periods than were the RPMI 1640-based media . Aminopterin supplemented medium was shown to be significantly less effective in supporting hybridoma reproduction and viability than medium without aminopterin . Antibody levels were directly related to cell concentration and viability regardless of the medium used for the hybridoma culture . An optimally formulated DMEM-based medium is suggested as the medium of choice for hybridoma propagation and maintenance. J In Vitro Fert Embryo Transf, 1988 Feb, 5(1), 25 - 30 Effects of prolactin on gametes and zygotes during in vitro fertilization in mice; Fukuda A et al.; Hyperprolactinemia is one of the major causative factors of infertility . However, the effect of prolactin on gametes during in vitro fertilization has not been elucidated . In the present study, the effects of mouse prolactin on the motility of spermatozoa, in vitro fertilization, and in vitro development of the zygote were investigated in mice using media containing three different concentrations (10, 50, and 100 ng/ml) of mouse prolactin . The development of unfertilized and fertilized oocytes (zygotes) was not affected in vitro by prolactin regardless of the amount of prolactin added to culture media . However, the fertilizing capacity of the spermatozoa was suppressed when they were preincubated for 90 min in culture media containing prolactin at concentrations of 50 and 100 ng/ml . The motility of spermatozoa was not affected by prolactin regardless of the concentration of prolactin used for preincubation . The present study indicates that prolactin may have some effects on the capacitation process of spermatozoa in vitro . This result should be taken into consideration in in vitro fertilization and embryo transfer procedures in humans. J Endocrinol, 1988 Feb, 116(2), 247 - 58 The time-course of oxytocin secretion from cultured bovine granulosa cells, stimulated by ascorbate and catecholamines; Luck MR et al.; Bovine granulosa cells secrete oxytocin when cultured in a serum-supplemented medium . The time-course of secretion is similar to that in the early corpus luteum in vivo, with a delay of 1 to 2 days followed by a peak and decline over the first 5 days of culture . We have investigated the basis of this time-course in vitro and studied the temporal characteristics of the stimulatory actions of ascorbic acid and adrenaline on this process . Cells cultured on stirred microcarriers showed a similar pattern of secretion of oxytocin to those cultured on conventional flat plates, despite continuing and rapid mitosis . This indicated that the secretion profile in conventional culture was not an artifact related to the cessation of mitosis . Furthermore, secretion of oxytocin and progesterone by cells on microcarriers was stimulated without a corresponding change in mitotic rate, showing that the secretion per cell had been increased . In conventional culture, addition of ascorbic acid to culture media (0.5 mmol/l) increased the secretion of oxytocin (up to 4.5-fold) but only if ascorbic acid was present during the first day of culture . The cells showed a progressive refractoriness to stimulation after 12 h . Since the time-course of secretion was unaltered by treatment, this resulted in a delay of 1 to 2 days before the action of the ascorbate was seen . The secretion of progesterone was similarly affected but with less stimulation and less consistency . In contrast, cells treated with adrenaline (10 mumol/l) secreted more oxytocin on the day of treatment and did so at any time during culture provided that there was sufficient basal secretion of hormone . Adrenaline also failed to alter the time-course of secretion but treated cells showed a persistent response, maintaining enhanced secretion for up to 3 days after the adrenaline had been removed . Ascorbate and adrenaline were highly synergistic in their effects, provided that the ascorbate was present from the start of culture; the response to adrenaline strongly reflected the degree of ascorbate stimulation . We conclude that granulosa cells secrete oxytocin according to an inherent time-schedule and that there is a limited period during which they can respond to ascorbate . Since ascorbate is required for the biosynthesis of oxytocin, this suggests that the availability of ascorbate during corpus luteum formation may determine the amount of oxytocin which can be released subsequently in response to catecholamines. J Clin Microbiol, 1988 Feb, 26(2), 383 - 4 UV red fluorescence of Veillonella spp; Brazier JS et al.; A total of 34 clinical isolates and 7 type strains of Veillonella spp . were tested for their ability to fluoresce on various culture media . Fluorescence was medium dependent and varied among the species . Scanning absorption spectrophotometry of culture extracts showed that the absorption spectrum of the fluorescent pigment is typical of a metal-free porphyrin. Radiat Res, 1988 Feb, 113(2), 243 - 51 Factors influencing the oxidation of the radioprotector WR-1065; Tahsildar HI et al.; N-(2-Mercaptoethyl)-1,3-diaminopropane (WR-1065) is the free thiol form of the radio- and chemoprotector S-2-(3-aminopropylamino)ethylphosphorothioic acid (WR-2721) . Interest currently exists in the clinical use of WR-2721 and WR-1065 as radio- and chemoprotectors of normal tissues . However, measurement of plasma levels of WR-1065 has proven difficult, due to rapid drug oxidation . Therefore, we studied factors influencing the oxidation of WR-1065, in Hepes-buffered saline as well as in tissue culture media containing 10% fetal bovine serum . The rate of oxygen consumption by WR-1065, as determined using the Clark oxygen electrode system, was faster in medium plus serum than in Hepes-buffered saline . That this effect is largely due to the presence of trace metal ions in tissue culture media and serum was indicated by the observation that addition of Cu2+ or Fe3+ to buffer stimulated oxygen consumption . Addition of KCN inhibited the reaction of WR-1065 with oxygen, and this effect was dependent on KCN concentration . That KCN blocked WR-1065 oxidation to the disulfide was verified using Ellman's reagent to quantitate the free thiol form . The rate of oxygen consumption was shown to be affected by temperature as well as concentration of WR-1065 . Catalase reduced the rate of oxygen consumption of WR-1065, indicating that peroxide is formed in this system . Superoxide dismutase had a stimulatory effect . WR-1065 was found to stimulate the hexose monophosphate shunt in A549 cells . Since this stimulation was prevented by the presence of catalase, it appeared to be due to the response of the cells to peroxide, formed as a result of WR-1065 autooxidation. Cryobiology, 1988 Feb, 25(1), 31 - 7 Skin preservation at 4 degrees C: a species comparison; Rosenquist MD et al.; There are numerous experimental studies in the literature regarding skin storage and preservation . These studies are difficult to interpret due to the variety of storage techniques utilized and the number of different animal species used as skin donors . This study utilized a single cold storage protocol to test the effect of species variation on skin graft viability . Donor skin was obtained from five animal species and human surgical panniculectomy specimens . The skin was stored in modified Roswell Park Memorial Institute (RPMI) 1640 tissue culture media at 4 degrees C . Stored skin was transplanted to surgically created defects on athymic (nude) mice after specific storage intervals . Ten days after transplantation, the grafts were examined by gross and microscopic techniques . The viability of mouse, rat, and dog skin was significantly different from human skin, while stored rabbit and pig skin were similar to human skin . These results demonstrate the difficulty of applying the data of skin storage studies from nonhuman species to clinical practice . The data indicate that rabbit and pig skin may be used in laboratory studies of skin preservation at 4 degrees C with a strong likelihood that the results may be of clinical relevance in predicting the behavior of human skin under similar storage conditions. Eur J Haematol, 1988 Feb, 40(2), 126 - 9 Kinetics of hemopoietic stem cells in a hypoxic culture; Ishikawa Y et al.; The influence of low oxygen tension on the clonal growth of hemopoietic stem cells in vitro was examined . The numbers of colonies of neutrophil, macrophage, and eosinophil progenitors (CFU-C), derived from human bone marrow, increased at a rate 1.7 times higher in low oxygen tension (7% O2) than in a gas phase that contained air (19% O2) . The erythroid (BFU-E) and multipotential (CFU-mix) progenitors increased about 2.4 times in 7% O2, and the increase in the composed cell type of mixed colonies showed no rate difference in either gas phase . Under atmospheric conditions, a mouse mast cell progenitor (CFU-mast) formed colonies, with the addition of 2-mercaptoethanol (2-ME) . Under low oxygen tension, the CFU-mast formed colonies without 2-ME, but a further enhancement was observed with the addition of 2-ME . Blood gas analysis of human bone marrow showed a pO2 of 51.8 +/- 14.5 mmHg, which was closed to O2 tension in a gas phase culture media containing 7% O2 . This data shows that the physiological O2 tension enhances hemopoietic stem cell proliferation in vitro, and that a part of the enhancing effect by 2-ME is due to a prevention of O2 toxicity at 19% O2. Prostaglandins Leukot Essent Fatty Acids, 1988 Feb, 31(2), 73 - 81 Effect of phorbol ester on prostaglandin regulation of proliferation in rabbit endometrial cells; Orlicky DJ et al.; We have proposed that two of the endogenously synthesized endometrial prostaglandins, prostaglandin F2 alpha (PGF2 alpha) and prostaglandin E1 (PGE1), play a regulatory role in growth control of the endometrium . PGF2 alpha increases DNA synthesis and PGE1 inhibits that effect . Primary cultures of rabbit endometrial cells were used here to examine the effects of the tumor-promoting, diacylglycerol mimicking, phorbol ester, 12-O-tetradecanoyl phorbol-13-acetate (TPA), on the prostaglandin control of cell proliferation . TPA treatment of these cultures results in: a decrease in control levels of proliferation and complete inhibition by TPA of PGF2 alpha stimulated DNA synthesis; a reduction in {3H}PGF2 alpha binding with short term treatment but an increase to above control binding level with long term treatment; an inhibition of the normal PGF2 alpha stimulated inositol polyphosphate synthesis; and a small increase in accumulation of PGF2 alpha in the culture media . Furthermore, in this culture system, TPA does not down regulate {3H}PGE1 binding; it does not alter the normal PGE1 stimulation of cAMP synthesis; and it has no effect on the normal endogenous PGE1 synthesis by these cultures . The above results are consistent with our previous observations that PGF2 alpha works through the intracellular messengers inositol polyphosphate/diacylglycerol whereas PGE1 works through cAMP. J Immunol Methods, 1988 Jan 21, 106(1), 57 - 61 A turbidometric assay for measuring proteins in culture media; Wojciechowicz D et al.; An immunoturbidometric assay was developed for the measurement of proteins in culture fluids of hepatocytes . The assay is simple to perform and avoids the biohazards associated with radioimmunoassays . The limit of detection of this assay exemplified by hemopexin and transferrin is 5 ng/ml protein . This degree of sensitivity is attained by incorporating into the procedure the addition of polyethylene glycol to enhance formation of primary antigen-antibody complexes and of a second antibody to further increase the immune complex size, which favors the ratio of specific to background light scattering. Thromb Res, 1988 Jan 15, 49(2), 287 - 97 Progestogen regulation of tissue plasminogen activator in a human melanoma cell line; Kjaeldgaard A et al.; The influence of progestogens, i.e . medroxyprogesterone, levonorgestrel and norethisterone, was studied in a melanoma cell line producing tissue-type plasminogen activator (t-PA) . The cell cultures were exposed to the three progestogens by addition of the steroids dissolved in a weak alcoholic solution to the culture media, in which the released t-PA was assayed by an immunoradiometric method . Ethanol (0.76% w/v) stimulated the t-PA production, while significant inhibitory effect of the present progestogens in the concentration of 1.7 x 10(-6) M was recorded . By tenfold decrease in progestogen concentration significant reduction of t-PA levels was still seen in the cultures exposed to levonorgestrel, while medroxyprogesterone and norethisterone in this dose had no effect on t-PA release . Norethisterone differed from the other two progestogens in having weak toxic effect on melanoma cells . It was concluded that the progestogens studied, in particular norethisterone and levonorgestrel, had an inhibitory effect on the production of t-PA in melanoma cell culture. Vopr Onkol, 1988, 34(7), 832 - 9 {Structural differences of the supramolecular DNA complex from tumor strain cells}; Blokhin DIu et al.; Elastoviscosimetric examination was carried out to study the structure of supramolecular DNA complexes obtained from transplantable tumor strain cells and those of the same strains showing pronounced growth in synthetic culture media . The supramolecular DNA complex was isolated as a nucleoid and examined by ethidium bromide elastoviscosimetric titration method . Levels of supercoiled DNA were found to be significantly lower in the chromatin of cells of in vitro growing strains, as compared to parental strain cells where the said type of DNA predominated . Two patterns of DNA compact packing--supercoiled-domain and one characterized by a faster bond between DNA and non-histone proteins, and single-stranded DNA areas preexisting in domains--were suggested. J Hyg Epidemiol Microbiol Immunol, 1988, 32(1), 79 - 85 Investigation of hemolytic activity of leptospirae on solid culture media; Volina EG et al.; Results of investigation of hemolytic activity of leptospirae against red blood cells of various animal species on solid culture media using the technique of agar layers are presented . It has been shown that hemolytic properties of pathogenic and saprophytic leptospirae differ with respect to red blood cells of the sheep, the rabbit, the hamster, the albino rat and the albino mouse . Hemolytic activity of leptospirae regarding the erythrocytes of the cattle, the horse and the fowl depends on the strain of leptospirae . The advantage of using solid nutrient media for the determination of hemolytic properties of leptospirae in comparison with liquid media has been demonstrated. Diagn Microbiol Infect Dis, 1988 Jan, 9(1), 7 - 10 Discrepancy between growth of Coccidioides immitis in bacterial blood culture media and a radiometric growth index; Ampel NM et al.; Spherules of Coccidioides immitis grew readily after inoculation in vented trypticase soy broth, biphasic brain heart infusion media, and aerobic tryptic soy broth bottles used in a radiometric system (BACTEC) . However, visible growth was not accompanied by a significant radiometric growth index . Growth of C . immitis can be visually detected in routine bacterial blood culture media while the radiometric growth index remains negative. Free Radic Biol Med, 1988, 4(2), 79 - 83 Quantitation of intracellular oxidation in a renal epithelial cell line; Scott JA et al.; We quantitated the presence of intracellular oxidizing species in response to oxidative stimuli using fluorescent cell analytic techniques . The studies were performed with a laser-activated flow cytometry system using 2',7'-dichlorofluorescin diacetate (DCFDA) as a probe for intracellular oxidation events . Oxygen radical formation was initiated by the addition of FeCl2 or xanthine oxidase to the culture media . Xanthine oxidase and FeCl2 both increased intracellular DCFDA oxidation over control (p less than .001) . Increases in intracellular DCFDA oxidation in response to xanthine oxidase exposure were inhibited by extracellular superoxide dismutase, catalase and dimethyl sulfoxide (p less than 0.001), implicating the superoxide anion, hydrogen peroxide, and the hydroxyl radical in producing the changes in intracellular dichlorofluorescein fluorescence . Increases in intracellular DCFDA oxidation in response to xanthine oxidase correlated with loss of cellular viability, as established by decreased plating efficiency . We conclude that relative intracellular oxidation can be quantitated within the cultured renal cell and that some extracellularly generated radicals may be capable of traversing the intact cell membrane to oxidize DCFDA in the cell interior. J Orthop Res, 1988, 6(2), 259 - 64 Time-varying magnetic fields: effects of orientation on chondrocyte proliferation; Elliott JP et al.; The purpose of this study was to determine the effect of orientation of pulsed electromagnetic fields (PEMFs) on cellular proliferation and extracellular matrix synthesis . Bovine articular chondrocytes were cultured in PEMFs (repetitive pulse at 72 Hz) generated using Helmholtz coils oriented either parallel (horizontal) or perpendicular (vertical) to the plane of cell adhesion . Dissipation of signal energy in the form of heat increased the temperature of the PEMF coils by 2 degrees C and the tissue culture medium by 1 degree C . Therefore, control coils, which emitted no PEMFs, were heated to the temperature of PEMF coils by circulating water . Chondrocytes were cultured in 16-mm-well culture plates, and the data for individual wells were pooled as triplicates . Although not observed by microscopic examination of individual wells, positionally dependent electric field effects may be minimized by this approach . PEMFs generated by coils oriented vertically significantly decreased chondrocyte proliferation . The effect was dependent on the concentration of serum in the culture media . At 3% serum concentration, the total cell number attained after 10 days of culture was reduced by 50% in stimulated cultures when compared with controls . At 5% serum concentration, there was no effect . PEMFs applied by coils oriented horizontally did not alter proliferation of articular chondrocytes . PEMFs had no effect on synthesis of extracellular matrix by chondrocytes plated at high density, irrespective of orientation. J Lab Clin Med, 1988 Jan, 111(1), 28 - 34 Complement fixation by pemphigus antibody . IV . Enhanced epidermal cell detachment in the absence of human plasminogen; Doubleday CW et al.; It has been reportedly previously that complement enhances pemphigus vulgaris antibody-induced epidermal cell detachment . The present studies were designed to eliminate the possibility of the human plasminogen-plasmin system contributing to the cell detachment observed in previous complement-mediated studies . To assay for the presence of protease contamination in the reagents, a simple, sensitive, caseinolytic-fluorometric assay was used . Confluent murine or human epidermal cell cultures were incubated with pemphigus IgG with and without complement and plasminogen . Detached epidermal cells, as determined by light microscopy and cell-counter analysis, were enumerated at 48 hours . Incubating pemphigus IgG with complement resulted in marked cell detachment, compared with minimal cell detachment when pemphigus IgG was incubated with plasminogen . Culture media were collected after 48 hours and assayed for activation of plasminogen . It was determined that the added plasminogen had not been activated, particularly in experiments where significant detachment was observed . The plasminogen could still be activated by streptokinase and urokinase . These studies suggest that epidermal cell detachment in pemphigus appears to be mediated at least in part by complement activation. Fertil Steril, 1988 Jan, 49(1), 108 - 11 Endotoxins in culture medium for human in vitro fertilization; Fishel S et al.; Endotoxins were detected in a few batches of culture medium during the authors' human in vitro fertilization program . Two distinct levels of endotoxins were assayed: greater than 1 ng/ml and less than 1 ng/ml . The source of endotoxin was traced to culture media obtained from a reputable manufacturing company . The incidence of fertilization per patient was not significantly affected by the presence of endotoxins, but fertilization assessed on the overall number of oocytes was significantly reduced (53%) when endotoxin levels were greater than 1 ng/ml compared with an assay negative for endotoxins (66%) (P = 0.047) . The percentage of oocytes cleaving after the observation of two pronuclei was not significantly different, but the degree of fragments observed in the conceptus was significantly more severe if the endotoxin level reached 1 ng/ml . In this investigation, the incidence of pregnancy was 8% when the endotoxin level was greater than 1 ng/ml, 30% if less than 1 ng/ml, and 32% if no endotoxins were detected . This study suggests that, although endotoxins may be present in the culture medium at a deleterious level of at least 1 ng/ml, fertilization and cleavage will be obtained, but there will be a significant increase in the incidence of conceptuses with cytoplasmic fragments; this may result in a reduction in the incidence of pregnancy. J Burn Care Rehabil, 1988 Jan-Feb, 9(1), 52 - 4 Short-term skin preservation at 4 degrees C: skin storage configuration and tissue-to-volume medium ratio; Rosenquist MD et al.; This study was designed to examine the effect of the storage configuration of skin and the ratio of tissue-to-storage medium on the viability of skin stored under refrigeration . Human skin specimens were stored in four physical configurations in RPMI 1640 tissue culture media at 4 degrees C . Skin was transferred to surgically created defects on nude mice after specific storage intervals . Grafts were examined grossly and microscopically after ten days . In the rolled configuration, on storage day 15, the viability of the outside of the roll was significantly better than the inside (P less than 0.01) . The graft viability of the outside of the skin rolls was similar for both tissue-to-media ratios as well as for both free-floating configurations (P = 0.27) . These findings suggest the optimum cold storage configuration is free floating, and 300 cm2/100 mL is an appropriate skin surface area to volume media ratio . This proportion of tissue to media is in agreement with the minimum ratio currently recommended by the Skin Council of the American Association of Tissue Banks. Aust J Biol Sci, 1988, 41(2), 189 - 99 Development of sheep embryos in vitro in a medium supplemented with different serum fractions; Batt PA et al.; Sheep embryos will generally develop into expanded blastocysts in vitro only in culture media supplemented with serum or serum components . In order to better understand how serum supports embryo development, a batch of ovine serum was fractionated by (a) ultrafiltration into two components containing substances with molecular weights greater and less than 10 Kd (kilodaltons), and (b) gel filtration into protein fractions 1, 2 and 3, containing groups of proteins with mean molecular weights of about 500, 150 and 65 Kd, respectively . The principal protein in fraction 3 was albumin . Day 6 sheep morulae were cultured in vitro for 48 hours in a bicarbonate-buffered salt solution supplemented with various concentrations of ovine serum or of these components or protein fractions of serum . Morulae could develop to fully expanded blastocysts in medium supplemented with whole serum or with the greater than 10 Kd component or protein fraction 3 only, but could not develop in medium supplemented with the less than 10 Kd component only or with the less than 10 Kd component and protein fractions 1 or 2 . However, the proportion of embryos that developed fully in medium supplemented with the greater than 10 Kd component or protein fraction 3 was increased by adding the less than 10 Kd component of serum to the medium . The addition of protein fraction 2 decreased the proportion of embryos that developed to expanded blastocysts in medium containing protein fraction 3 and the less than 10 Kd component, but not in medium containing whole serum . Since the compositions of different sera may vary markedly, these results suggest (a) reasons why different sera vary in their ability to support embryo development in vitro, and (b) factors which may influence development of the sheep embryo in the uterus, where plasma proteins comprise nearly all the protein in the fluid bathing the embryo. Reprod Nutr Dev, 1988, 28(6B), 1791 - 5 {Search for specific factors in culture media of embryos}; Testart J et al.; Mitogenic activity of culture media having contained either unfertilized or cleaved embryos have been studied . In these media human eggs have been cultured for one day, beginning 20 hours following in vitro insemination . B-lymphocyte proliferation was estimated by thymidine incorporation following 3 day culture in the presence of eggs or embryo culture media . No reproducible effect on B lymphocyte proliferation was observed . Therefore the viability of embryos could not be ascertained by the mitogenic activity of their culture media. J Immunoassay, 1988, 9(2), 193 - 206 A simple radioimmunoassay of human interleukin-2; Lee TP et al.; We have developed a liquid phase radioimmunoassay of human interleukin-2 which uses inexpensive commercially available reagents exclusively . The assay is simple, reproducible and specific in detecting different batches of human interleukin-2 of natural as well as recombinant origin, but not detecting recombinant murine interleukin-2 . The assay is sensitive to a concentration of approximately 0.05 ng/ml and can be used in measurement of IL-2 in serum containing culture media. Acta Biol Hung, 1988, 39(1), 75 - 85 Effect of heavy metals on the growth of tissue cultures (II); Maroti M et al.; The effect of toxic concentrations of three heavy metal compounds on the growth of the secondary callus tissue of Nicotiana tabacum L . and Ruta graveolens L . was studied . The metal compounds examined were ZnSO4, NiSO4, CuSO4 . The metal compounds used were placed in Murashige, Skoog (1962) and White (1943) culture medium at 10(-6) and 10(-4) M concentration, respectively, before autoclaving . The culture media containing macro- and microelements and vitamins were completed with carbon source and regulators (IAA, GA, kinetin for Nicotiana and IAA, 2, 4-D for Ruta) . The cultures were kept for 4 weeks at 25 (+2) degrees C under 16/8 n light/dark conditions . The value of pH was 5.6 before the autoclave treatment . The increase in fresh weight of the secondary callus tissue was inhibited by the metal compounds applied with both plant species (to 75-87% by zinc, 7-97% by nickel, 5-98% by copper with tobacco; to 47-69% by zinc, 5-88% by nickel, 57-90% by copper with rue) . The cell number and dry weight per g of callus tissue partly increased, partly decreased compared to the control in response to the heavy metal treatment . The growth values obtained with various concentrations of the heavy metals were different in the two plant species due to differences in metabolism and organization potential between them. Tissue Cell, 1988, 20(4), 541 - 54 Primary culture of bovine mammary acini on a collagen matrix; Baumrucker CR et al.; Lactating bovine mammary epithelial acini were isolated and primary culture on rat tail attached collagen gels are described . Acini rapidly attach to the gels and morphologically change little over days of culture under the culture conditions described herein . Cells release lactose, alpha-lactalbumin and alpha-s1 casein over a 6-day period . A new HPLC method for measuring lactose in mammary cell culture media is described . Comparisons of acini cultures with individual cell cultures show acini to be 1.5-5 times more productive than cells in secreting lactose and casein, respectively. Cell Tissue Res, 1988, 254(3), 611 - 5 Monoclonal antibody C1B8A8 recognizes a ventricular secretory product elaborated in the bovine subcommissural organ; Meiniel R et al.; To obtain specific immunological probes for investigation of the cellular and molecular aspects of the subcommissural organ (SCO), we produced monoclonal antibodies directed against extracts from the bovine SCO . An hybridoma cell line (C1A8B8) was isolated by screening the culture media by means of the immunofluorescence method . This clone produces an IgG1 that recognizes the ventricular secretory material of the SCO including Reissner's fiber . A competition test using C1B8A8 immunoglobulin and lectins (concanavalin A and wheat-germ agglutinin) was applied to demonstrate that both the immature and mature forms of the glycoprotein were recognized . This antibody will offer a good tool for immunocytochemical localization and immunoaffinity purification of the antigen and for isolation of cDNA clones encoding it. Pharmacology, 1988, 37(3), 154 - 64 Cell-catalyzed binding of 3H-(-)-trans-7,8-dihydroxy-7,8-dihydrobenzo{a}pyrene to cellular and exogenous DNA and the role of purified human liver epoxide hydrolase; Gozakara EM et al.; Cultured human monocytes, lymphocytes, Fischer rat liver (TRL-2) cells, and Buffalo rat liver (BRL) cells catalyzed the conversion of 3H(-)-trans-7,8-dihydroxy-7,8-dihydrobenzo{a}pyrene {3H(-)t-7,8-dihydrodiol BP} to r-7,t-8-dihydroxy-t-9,10-oxy-7,8,9,10-tetrahydrobenzo{a}pyrene (diol epoxide I) and r-7,t-7-8-dihydroxy-c-9,10-oxy-7,8,9,10- tetrahydrobenzo{a}pyrene (diol epoxide II; r-7 indicates that the substituent at the 7-position is the reference, and t and c indicate that the substituents trans and cis, respectively, to the reference substituent) . These appear to be the most reactive metabolites of benzo{a}pyrene (BP) and were covalently bound to both exogenous and intact cellular DNA in tissue culture media . The cells induced by benzanthracene (BA) exhibited greater levels of DNA binding than the controls and this binding was linear with increasing cell content in human monocytes, in TRL-2 cells and in Buffalo rat liver cells . The binding to DNA was greater than controls in BA-preinduced lymphocytes, but was not linear . The DNA binding in control cells showed a nonlinear increase with increasing cell concentration in all experiments . The addition of human liver epoxide hydrolase (EC 3.3.2.3) to the incubation medium reduced the amount of reactive metabolites binding to DNA by 12-15% in control and by 23-41% in BA-induced monocytes . Thus, with whole cell systems of either human monocytes or lymphocytes, the addition of purified human liver epoxide hydrolase reduced the binding of 3H(-)t-7,8-dihydrodiol BP metabolites to DNA . Human monocytes and lymphocytes also catalyzed the covalent binding of 3H(-)t-7,8-dihydrodiol BP to intact cellular DNA . The addition of 3H(-)t-7,8-dihydrodiol BA to tissue culture media caused the inhibition of covalent DNA binding in BA-preinduced monocyte by 58% and lymphocytes by 25% . Previous work has shown that BA is metabolized and converted to BA-diol epoxides by microsomes . These results indicate that BA-diol epoxides and BP diol epoxides are competing for the same binding sites on DNA . On the other hand, the addition of 10 nmol of 3H(-)t-7,8-dihydrodiol BP to the incubation of control and BA-preinduced cell homogenate and further incubation at 37 degrees C for 25 min showed that the DNA binding in BA-preinduced cell homogenates was much greater than controls . Homogenates of cells induced by BA exhibited a greater level of DNA binding than controls.(ABSTRACT TRUNCATED AT 400 WORDS) Exp Pathol, 1988, 33(3), 173 - 7 Ethanol damage to rat gastric mucosa is unlikely to be mediated by ethanol; Sewell RB et al.; During injury to the gastric mucosa, lysosomes become more fragile and lysosomal enzymes, which are activated at acid pH, leak into the surrounding environment . It is not clear whether these changes contribute to the mechanism of damage or are merely a secondary result of it . To test whether lysosomes modulate gastric mucosal damage, we pretreated rats with a lysosomal labilizing agent, Triton WR 1339 (1.5 g/kg) and histologically assessed mucosal damage in vivo after challenge with 30% ethanol . No significant differences were found in the length or depth of eroded mucosa: mean erosion length, Triton 23.9 +/- 6.6% vs . control 19.7 +/- 5.2%; mean depth (micron), Triton 19 +/- 4 vs . control 20 +/- 7 . After a similar pretreatment regimen, rat antral mucosa was cultured, challenged with ethanol and damage assessed by release into media of previously incorporated mucosal 51chromium . With 15% ethanol challenge, no change in 51chromium release was seen: after Triton, 9.8 +/- 1.4% vs . control 10.3 +/- 1.0% . Triton pretreatment perturbed gastric lysosomes as shown in organ culture by significantly raised tissue lysosomal enzyme activities and increased lysosomal enzyme release into culture media after ethanol challenge . The lack of effect of this pretreatment regimen suggests that lysosomes do not have a major pathogenetic role in ethanol-induced gastric damage. Life Sci, 1988, 43(13), 1069 - 77 Characterization of 1,1-dimethyl-4-phenylpiperazinium induced increased proenkephalin processing in bovine chromaffin cells; Cherdchu C et al.; Acute stimulation of bovine adrenal chromaffin cells in culture with 1,1-dimethyl-4-phenylpiperazinium (DMPP) gives rise to a significant increase in secretion of {Met5}-enkephalin immunoreactive material (ME-IRM) into the culture medium (1) . Following this secretion the cellular ME-IRM levels do not decrease, suggesting the replenishment of the peptides . The repletion of the cellular ME-IRM appears to result from an increase in processing of large molecular weight peptides containing {Met5}-enkephalin and {Leu5}-enkephalin . Gel filtration chromatography on Bio-Gel P-10 was used to fractionate the enkephalin-like peptides (ELPs) present in the culture media and chromaffin cell extracts . Fractionation was done for samples before and after nicotinic receptor stimulation by DMPP to demonstrate the secretion and repletion of the ELPs . Gel chromatographic profiles of ELPs present in the culture media after DMPP stimulation revealed the presence of 4 peaks, representing different molecular forms of these peptides (Peaks 1-4), with a selective increase in secretion of Peaks 3 and 4 . The chromatograms of ELPs extracted from cultured chromaffin cells showed similar patterns to those obtained from ELPs present in the culture medium after stimulation . Analyses of individual peaks after fractionation of cell culture extracts showed an increase in the amount of immunoreactive material found in Peak 4 with a concomitant decrease in the immunoreactivity found in the higher molecular weight peaks (Peaks 1-3) . Further purification of Peak 4 from cell extracts on reversed-phase HPLC (RP-HPLC) showed a significant amount of ELPs existed as the sulfoxide derivative of {Met5}-enkephalin . The content of {Met5}-enkephalin sulfoxide (ME-O-enk) did not decrease following DMPP stimulation . We conclude that acute stimulation of nicotinic receptors in the chromaffin cells enhances the processing of proenkephalin precursors to keep pace with the secretion of low molecular weight peptides. Leuk Res, 1988, 12(7), 567 - 74 Haemoglobin synthesis in K562 erythroleukaemia cells is affected by intimate contact with monolayers of various human cell types; Zuhrie SR et al.; The haemoglobin content of K562 erythroleukaemia cells was affected by co-culture over monolayers of various human cell types . Haemoglobin synthesis was increased after co-culture with umbilical-cord-derived endothelial cells and most monolayers of bone-marrow-derived macrophages, and inhibited after co-culture with two fibroblast lines, blood-monocyte-derived macrophages, a neuroglial cell line (U-251 MG) and most monolayers of bone-marrow-derived stromal cells . These effects were modified when a thin layer of agar was placed over the monolayers . Cell-free culture media conditioned by all but two of the seven types of monolayer studied inhibited haemoglobin synthesis by K562 cells; those conditioned by blood-monocyte-derived macrophages and two of 11 monolayers of bone-marrow-derived macrophages stimulated haemoglobin synthesis . Thus, the haemoglobin content of K562 cells appeared to be influenced both by intimate contact between K562 cells and the cells of the monolayers and by humoral factors released by the monolayers . The data support the concept that erythroid differentiation is partly dependent on intimate contact between erythroid progenitor cells and microenvironmental cells. Int J Biochem, 1988, 20(8), 817 - 22 Inhibition of plasminogen activators and the growth of cultured human tumour cells; Scott GK; 1 . Inhibition of a human fibroblast cell-surface proteinase with alpha-1-antitrypsin also inhibited cell proliferation, confirming earlier observations . 2 . Although a similar proteinase inhibition could be observed with human tumour cells, the growth of these cells was unaffected . 3 . Plasminogen activators on the cell surface and in culture media could be inhibited with specific antibodies, but there was no corresponding effect on the growth of any of the cell types tested. Eur J Gynaecol Oncol, 1988, 9(3), 222 - 7 Effect of alpha-difluoromethylornithine (DFMO) on the growth of human ovarian carcinoma; Manetta A et al.; The antiproliferative effect of alpha-difluoromethylornithine (DFMO) was investigated in a human ovarian cancer cell line (NIH:OVCAR3) both in vitro and in vivo . DFMO at 5 X 10(-5) M, 5 X 10(-4) M and 1 X 10(-3) M concentrations showed growth inhibition of NIH:OVCAR3 cells in culture . Parallel determination of CA 125 in the culture media of these cells show significant decrease in the presence of DFMO compared to controls . Oral administration of DFMO (2% aqueous solution) to nude mice bearing intraperitoneal tumors resulted in a mean survival of 45 days (38-60) versus 25 days (20-35) for control . Both in vitro and in vivo results suggest that DFMO has potential value in the treatment of ovarian carcinoma and should be considered for clinical trials in appropriate cancer patients. Am J Trop Med Hyg, 1988 Jan, 38(1), 111 - 7 Murine schistosomiasis: selective inhibition in vitro of fibroblast collagen production by mononuclear cells; Mansour MM et al.; Primary cell cultures from the livers of mice infected with Schistosoma mansoni were prepared and cells with the appearance of fibroblasts by light microscopy were isolated . Collagen synthesis was estimated by measuring incorporation of 14C-proline into collagenase-sensitive proteins for both culture media and cell layers . Coculture of splenic T cells from infected mice with these hepatic fibroblasts caused greater selective and specific reduction in collagen production than did coculture using spleen cells from normal mice . There was a parallel inhibition in collagen within the cell layer which indicates that the marked decrease in collagen production was due to inhibition of synthesis and not related to changes in solubility or secretion . Primary culture of mouse skin fibroblasts showed similar responses to coculture but an established fibroblast line, 3T3, was unresponsive . Inflammatory cells appear to influence hepatic fibroblasts isolated under our experimental condition in several ways, such as opposite effects on collagen synthesis and cell proliferation. Ital J Surg Sci, 1988, 18(3), 201 - 5 The stimulating effect of a cytosol extract from regenerating liver on isolated hepatocytes and the positive role of insulin; Gorini P et al.; Several kinds of damage induce a regenerative response in the liver . Partial hepatectomy is a well known method, suitable for studying liver regeneration and factor/s involved in it . The presence of the so called Hepatic Stimulator Substance has been demonstrated in the cytosol obtained from regenerating hepatocytes, and it has been partially characterized, from a molecular point of view . Mainly, it stimulates liver regeneration and Tritiated Thymidine incorporation in cultured hepatocytes as demonstrated in this paper (Control Hepatocytes: 8447 +/- 660.4 cpm, (mean +/- sd), Test Hepatocytes: 16974 +/- 2720.9 cpm; Index of Stimulation: 2.01) . Furthermore, the stimulating activity of HSS is increased by the presence, in the culture media, of insulin which has been proved to have a positive role in liver regeneration (Control Hepatocytes: 8889 +/- 367.3 cpm; Test Hepatocytes: 21588.5 +/- 1232.4 cpm; Index of Stimulation: 2.42, p less than 0.05) . These data seem to suggest that liver is probably the site of production/activity and action of factor/s that stimulate/s liver regeneration; but other substances exist, such as insulin, which play a permissive role. Biol Met, 1988, 1(1), 9 - 17 High-performance liquid chromatography of siderophores from fungi; Konetschny-Rapp S et al.; A reversed-phase HPLC separation of iron(III) chelates of 16 representative fungal siderophores including ferrichromes, coprogens and triacetylfusarinine C was established in order to investigate siderophore production of fungi . For comparison purposes, the widely used bacterial siderophore ferrioxamine B was included . Culture filtrates of the fungi Penicillium resticulosum, Fusarium dimerum, Aspergillus fumigatus and Neurospora crassa were quantitatively analyzed for the presence of known and unknown siderophores after growth in low-iron culture media and adsorption on XAD-2 columns using this HPLC separation system . Photodiode array detection allowed the distinction between siderophores and non-siderophores . According to their ultraviolet/visible spectra, a further classification of the siderophores into four types due to the number of anhydromevalonic acid residues per molecule (0-3) was possible. J Hosp Infect, 1988 Jan, 11(1), 77 - 81 Isolation of organisms in CAPD peritonitis: use of nutrient broth cultures and Bactec blood culture media; Stokely DJ et al.; Cultures of the first cloudy dialysates from 51 consecutive episodes of peritonitis in patients undergoing continuous ambulatory peritoneal dialysis were carried out using concentrated nutrient broth and Bactec blood culture media in addition to primary culture on solid media . Thirty-seven episodes (73%) were positive on solid media, and 42 episodes (82%) were positive by both nutrient broth and Bactec methods . The value of Gram-stained smears and WBC counts on dialysates is discussed. Drugs Exp Clin Res, 1988, 14(1), 19 - 23 Itraconazole: increased activity by chlorhexidine; Simonetti N et al.; Chlorhexidine increases the activity of itraconazole against Candida isolates; itraconazole-chlorhexidine combinations show synergistic activity in culture media . The activity of itraconazole is discussed. J Orthop Res, 1988, 6(4), 525 - 30 The synovial production of collagenase and chondrocyte activating factors in response to cobalt; Ferguson GM et al.; Addition of CoCl2 solutions to the culture media of confluent monolayers of lapine or human synoviocytes stimulated their production of the neutral proteinases collagenase, gelatinase, and caseinase . With lapine cells, maximum stimulation occurred at 10(-7) M CoCl2, while human cells required 10(-4)-10(-5) M CoCl2 to achieve a maximum stimulation . Production of prostaglandin E2 by lapine cells was enhanced some 30-40% by concentrations of CoCl2 that maximally stimulated synthesis of the neutral proteinases, whereas all concentrations of CoCl2 slightly depressed the production of prostaglandin E2 by human cells . Lapine synovial cells that had been stimulated by CoCl2 also produced a substance, or substances, that provoked the synthesis of collagenase, gelatinase, caseinase, and prostaglandin E2 by monolayers of articular chondrocytes . Chondrocytes themselves, however, resisted activation by CoCl2 . These findings may be relevant to the aseptic loosening of joint prostheses. Drugs, 1988, 35 Suppl 1, 33 - 41 Effects of interleukin-1 and anti-inflammatory drugs on the degradation of human articular cartilage; Shinmei M et al.; It has been suggested that metalloproteases produced by chondrocytes play an important role in cartilage breakdown in joint diseases . The aim of this study was to investigate changes in enzyme activities in human rheumatoid and osteoarthritic articular cartilage . Cartilage fragments were incubated with various drugs for 48 hours . The concentrated culture media were used as enzyme solutions . Collagenase was assayed using FITC-collagen as the substrate . Proteoglycanase (PGase) was measured either by the release of 35S-labelled proteoglycans from cartilage into the medium, or by enzyme assay using proteoglycan monomer bound to fluorescein-conjugated hyaluronic acid as the substrate . Collagenase and proteoglycanase were found only in trace amounts in the concentrated media of healthy cartilage . Interleukin-1 (IL-1) enhanced the enzyme activities significantly . Marked increases of enzyme activities were observed in the concentrated media of rheumatoid (RA) and osteoarthritic (OA) cartilage . The sensitivity to interleukin-1 was also higher in OA and RA cartilage compared with healthy cartilage . Dexamethasone (10(-6) mol/L) markedly depressed enzyme activity . Tiaprofenic acid (4 x 10(-5) mol/L) also decreased enzyme activity, whereas indomethacin (4 x 10(-6) mol/L) and naproxen (3 x 10(-4) mol/L) had no effect. Transfusion, 1988 Jan-Feb, 28(1), 81 - 3 Photodynamic therapy of viral contaminants with potential for blood banking applications; Matthews JL et al.; A photodynamic method has been evaluated as a means of eradicating viral contaminants with the potential for rendering blood safe for transfusion . Herpes simplex virus type 1 (HSV-1) was tested under flowing conditions in culture media or in blood supplemented with the virus . Hematoporphyrin derivative was used as the sensitizer and was photoactivated with visible light at 630 nm and 5 J/cm2 . HSV-1 in suspension both in culture medium as well as in blood was shown to be killed . The human immunodeficiency virus was also found to be photoinactivated in flowing cell culture medium and, thus, potentially may be inactivated in blood . These findings extend our previous studies which demonstrated that enveloped viruses can be photoinactivated with hematoporphyrin derivative in a static fluid system . Analysis of blood cell number, red cell lysis, plasma proteins, and other standard hematological tests showed no significant change . The possibility that transfusion-associated acquired immunodeficiency syndrome (AIDS) may result from a blood unit infec