Cell Differ, 1988 Aug, 24(3), 179 - 90 A monoclonal antibody recognizing a common antigen on neurons and fibroblasts in chicken and quail; Graniou M et al.; A monoclonal antibody, FiN1, obtained by immunization of a mouse with homogenates of embryonic quail nodose ganglia, was found to react with a surface antigenic determinant, both in quail and chick, present on practically all neurons of the spinal cord and of the peripheral nervous system and on a subpopulation of fibroblasts . An ontogenetic study performed on tissue sections, cell suspensions and cultures showed that FiN1 defines a differentiation marker which appears relatively late in development, during the second half of embryonic life, and persists after hatching . The onset and evolution of its expression during development varies in a tissue-specific manner. Biophys J, 1988 Aug, 54(2), 241 - 7 Hypophosphite ion as a 31P nuclear magnetic resonance probe of membrane potential in erythrocyte suspensions; Kirk K et al.; Hypophosphorus acid has a single pKa of 1.1 and at physiological pH values it is therefore present almost entirely as the univalent hypophosphite ion . When added to a red cell suspension the ion crosses the cell membrane rapidly, via the anion exchange protein, and the intra- and extracellular populations of the ion give rise to separate 31P NMR resonances . From a single 31P NMR spectrum it was possible to determine the relative amounts of hypophosphite in the intra- and extracellular compartments and thereby estimate the corresponding concentrations . The ratio of intracellular to extracellular hypophosphite concentration was independent of the total hypophosphite concentration for cells suspended in NaCl solutions and was independent of hematocrit . The hypophosphite distribution ratio increased as extracellular NaCl was replaced iso-osmotically with citrate or sucrose, through it remained very similar to the corresponding hydrogen ion distribution ratio . Incorporation of the hypophosphite distribution ratio into the Nernst equation yielded an estimate of the membrane potential . For cells suspended in NaCl solutions the estimated potential was consistently around -10 mV. Cell Biol Int Rep, 1988 Aug, 12(8), 637 - 46 Preliminary studies on the cultivation and characterization of mini-pig prostate epithelial cells; Costello LC et al.; Mini-pig prostate epithelial cells exhibited the unique metabolic characteristics associated with the specialized function of production and secretion of high levels of citric acid . Epithelial cell suspensions from mini-pig prostate were successfully grown in primary and secondary cultures . The cultured epithelial cells exhibited rapid proliferation reaching confluency in approximately 6 days . Growth and proliferation of fibroblasts were markedly restricted by the dominance of epithelial cell growth . Confluent cultures could be maintained for approximately 6 weeks . The epithelial cells retained their polymorphic appearance in primary and secondary cultures and exhibited the characteristic formalin-resistant acid phosphatase reaction . Testosterone stimulated mitochondrial aspartate aminotransferase (mAAT) activity and citrate production by confluent epithelial cell cultures . These initial results indicate that cultured epithelial cells derived from mini-pig prostate might be an excellent model related to human for studies of prostate biology and hormonal regulation. Neuroscience, 1988 Aug, 26(2), 417 - 31 Ultrastructural evidence for hippocampal target cell-mediated trophic effects on septal cholinergic neurons in reaggregating cell cultures; Hsiang J et al.; We have previously demonstrated at the light microscopic level that when embryonic day-15 septal neurons are co-cultured for 21 days with their target cells from the hippocampus, increased numbers of septal cholinergic neurons are present as compared with co-cultures employing cells from the non-target cerebellum . In addition, fine varicose axon-like cholinergic fibers are found to be associated with the hippocampal cells but not with cerebellar cells . We now provide ultrastructural evidence for hippocampal target cell-enhanced cholinergic neuronal survival, axonal proliferation, and synapse formation in this culture system . Dissociated cell suspensions from septal, hippocampal, and cerebellar areas were obtained from 15-day mouse embryos; and hippocampal and cerebellar cells were internally labeled with rhodamine-conjugated wheat germ agglutinin . Combinations of septal and hippocampal cells, and septal and cerebellar cells were allowed to reaggregate in rotation mediated culture for either 15 or 21 days . The reaggregates were then fixed, embedded, sectioned, and processed for acetylcholinesterase-positive acetylcholinesterase-positive cells and fibers, and under fluorescence to locate rhodamine-labeled cell populations . Representative reaggregate profiles were then re-embedded for electron microscopic examination . In both types of reaggregates, either labeled hippocampal target or cerebellar non-target cells segregated from the septal cells so that areas containing each of the respective cell populations could be studied . In sections of septal-hippocampal reaggregates from 15-day cultures, 571 out of 665 (85%) cholinergic neurons examined were intact, whereas 15% of the cells showed some ultrastructural features of degeneration . Similarly, at day 21, 297 out of 335 (88%) of the cholinergic neurons were intact . In sections of septal-cerebellar reaggregates from 15-day cultures, 473 out of 572 (83%) cholinergic neurons were intact . By day 21 of culture, however, only 15 out of 110 (14%) cholinergic neurons examined were intact from the septal-cerebellar reaggregates . In areas of septal-hippocampal reaggregates occupied by rhodamine-labeled hippocampal cells, profiles of acetylcholinesterase-labeled axons were identified, and synaptic specializations were observed between cholinergic terminals and dendrites as well as somata of hippocampal target cells . In contrast, areas of septal-cerebellar reaggregates occupied by rhodamine-labeled cerebellar cells were devoid of cholinergic fibers.(ABSTRACT TRUNCATED AT 400 WORDS) Arch Biochem Biophys, 1988 Aug 1, 264(2), 400 - 9 Alkaline phosphatase is an ectoenzyme that acts on micromolar concentrations of natural substrates at physiologic pH in human osteosarcoma (SAOS-2) cells; Fedde KN et al.; Alkaline phosphatase (ALP) was examined in cultured human osteosarcoma cells (SAOS-2) with respect to isoenzyme form, kinetic properties toward two natural substrates, and topography and nature of attachment to the plasma membrane . ALP in SAOS-2 homogenates is the tissue-nonspecific (TNS) isoenzyme and a phosphoethanolamine (PEA) and pyridoxal 5'-phosphate (PLP) phosphatase, as demonstrated by heat and inhibition profiles and electrophoretic mobility . Kinetic studies indicate that TNSALP in SAOS-2 cells has both a low- and a high-affinity activity . The high-affinity activity (showing the greater catalytic efficiency) is active at physiologic pH toward physiologic concentrations (microM) of PEA and PLP . TNSALP was shown to be an ectoenzyme in SAOS-2 cells by our findings in intact cell suspensions, where (i) PEA and PLP degradation in the medium nearly equaled that of whole cell homogenates, (ii) greater than 85% of ALP activity was inactivated by acid treatment, and (iii) ALP activity was quantitatively released by phosphatidylinositol-specific phospholipase C . Our findings indicate that, in SAOS-2 cells, TNS (bone) ALP functions as an ectoenzyme to degrade physiologic concentrations of extracellular natural substrates at physiologic pH. Cancer Res, 1988 Aug 1, 48(15), 4427 - 33 Activity of the human blood group ABO, Se, H, Le, and X gene-encoded glycosyltransferases in normal and malignant bladder urothelium; Orntoft TF et al.; Immunohistochemistry has led to the finding of an expression of ABO-related blood group antigens in normal and malignant bladder urothelium which is different from that found on erythrocytes from the same individual . This includes a loss of blood group ABO expression in malignant urothelium, and the expression of Leb antigens in normal and malignant cells from individuals with Le(a+b-) and Le(a-b-) erythrocytes . To elucidate the mechanism of this blood group antigen expression in urothelium we have analyzed the activity of the specific glycosyltransferases encoded by the ABO, Se, H, Le, and X blood group genes in normal and malignant urothelium . Biopsies of normal urothelium were obtained from 22 individuals and biopsies of urothelial tumors from 20 individuals . The tissue donors were typed for ABO, Lewis, and secretor status on erythrocytes and saliva . The biopsies were disaggregated to single cell suspensions, and the activity of the individual glycosyltransferases was determined as pmol of labeled sugar incorporated by oligosaccharide acceptors per 100,000 cells . The A (alpha-3-N-acetyl-D-galactosaminyl) and B (alpha-3-D-galactosyl) gene-specified transferases showed no activity in malignant cells, whereas all other enzymes examined were expressed in both normal and malignant cells . Secretors and nonsecretors showed the same alpha-2-L-fucosyltransferase activity in both normal and malignant cells, whereas the alpha-3-L-fucosyltransferase was reduced (P less than 0.02) in malignant cells from Lewis positive individuals . The Lewis gene-encoded alpha-4-L-fucosyltransferase showed a similar activity in Lewis positive and negative individuals . These results indicate that the disappearance of A and B blood group antigens in bladder tumors and the expression of Leb antigens in normal and malignant cells from individuals with Le(a+b-) and Le(a-b-) erythrocytes are due to corresponding differences in glycosyltransferases . The results indicate that the ABO, H, Se, and Le genes are subjected to a tissue-dependent differential expression. Am Rev Respir Dis, 1988 Aug, 138(2), 451 - 7 Bronchoalveolar lavage cell differential on microscope glass cover . A simple and accurate technique; Laviolette M et al.; We describe a quick and easy technique to perform cell differentials on bronchoalveolar lavage: the microscope glass cover . Lavage fluids of 72 subjects were analyzed by 3 techniques: glass cover, filter, and cytocentrifuge preparations . Seventy-seven other lavages were analyzed by glass cover and cytocentrifuge preparations alone . Data for the 72 subjects studied by all 3 techniques showed that the cell counts on glass cover and filter preparations were similar, e.g., lymphocytes, 19.2% (range, 0.5 to 94%) and 20.9% (range, 3 to 95%), respectively (Spearman's correlation coefficient, 0.98) . However, on cytocentrifuge preparations, lymphocyte counts were lower (8.3%; range, zero to 87%) and macrophage counts were higher (p less than 0.005) . Comparison of glass cover and cytocentrifuge preparation mixtures with varying amounts (20 to 80%) of purified blood leukocytes labeled with 51Cr (greater than or equal to 72% lymphocytes) showed that a significant amount of radioactive cells was lost during the cytocentrifuge technique in contrast to the glass cover technique . Because neutrophils represented a low proportion of lavage cells, we also evaluated cell suspensions with known neutrophil contents (10 to 70%); we found no difference in neutrophil counts obtained with the 3 techniques . Lavage data analysis of 40 young nonsmoking volunteers showed that glass cover lymphocyte count was also higher than counts on cytocentrifuge preparations: 16.5% (range, 3 to 45%) and 8.2% (range, 2.5 to 35%), respectively . In this group, the distribution of glass cover lymphocyte percentages was normal (p = 0.21, chi 2 test), and the one-tailed 95% confidence interval was 18.6 to 34.7% (mean plus 1.65 standard deviation).(ABSTRACT TRUNCATED AT 250 WORDS) Zh Mikrobiol Epidemiol Immunobiol, 1988 Aug, (8), 90 - 4 {Solid-phase immunoenzyme analysis of Coccidioides immitis antigens}; Khrapova NP et al.; A highly effective and specific solid-phase enzyme immunoassay system has been developed . The enzyme immunoassay is a highly sensitive technique for the detection and identification of C . immitis cellular and metabolic antigens . This technique is suitable for the study of strain differences in the antigenic composition of C . immitis, rendered harmless by different methods . The expediency of the preliminary sonification of cell suspensions of C . immitis, the causative agent of coccidioidomycosis, has been experimentally confirmed. Allergy, 1988 Aug, 43(6), 464 - 8 IgE on Langerhans cells in the skin of patients with atopic dermatitis and birch allergy; Tigalonowa M et al.; Epidermal cell suspensions from the skin of seven patients with atopic dermatitis (AD) and seven healthy non-atopic controls were investigated for the presence of surface HLA-DR and CD1 antigen, and IgE using indirect and double-staining immunofluorescence techniques . Fifty-seven percent of all CD1+ and 68% of all HLA-DR+ cells from the patients demonstrated IgE on their surface, indicating that Langerhans cells (Lc) in AD may be a heterogeneous population with regard to surface characteristics . No IgE+ cells were found in the epidermal cell suspensions from the healthy non-atopic controls . An attempt to demonstrate birch pollen antigen on the surface of Lc from the same patients all strongly allergic to birch pollen, using indirect immunofluorescence techniques, was unsuccessful, also after in vitro incubation of the Lc with high concentration of the birch antigen. Appl Environ Microbiol, 1988 Aug, 54(8), 2107 - 11 Influence of unsaturated fatty acid membrane component on sensitivity of an Escherichia coli fatty acid auxotroph to conditions of nutrient depletion; Massa EM et al.; The unsaturated fatty acid auxotroph Escherichia coli AK7 was provided with either oleic acid (cis 18:1) or linolenic acid (cis 18:3) to vary the degree of unsaturation of cell membrane lipids . The susceptibility of oleic acid- and linolenic acid-grown cells to starvation at 37 degrees C in 154 mM NaCl was compared following the decline in the number of CFU by plating the cells on agar medium . The decline in CFU was faster for linolenic acid-than for oleic acid-grown cells, but it was not indicative of cell death, since culturable CFU was recovered after respirable substrate was added to the starved cell suspension . Cell envelope microviscosity (determined by fluorescence polarization) of oleic acid- and linolenic acid-grown cells was equal in the presence of a respirable substrate, but in its absence the microviscosity of linolenic acid-grown cells was lower than that of oleic acid-grown cells . The results suggest that cell envelope microviscosity is an important factor in determining the sensitivity of E . coli to conditions of nutrient depletion. Exp Eye Res, 1988 Aug, 47(2), 269 - 82 Selective failure of long-term survival of isolated photoreceptors from both homozygous and heterozygous rd (retinal degeneration) mice; Politi L et al.; Retinas from homozygous rdle/rdle and heterozygous rdle/++ C57BL/6J mice were dissected and dissociated on postnatal day 2, when they are still essentially indistinguishable . The resulting cell suspensions were seeded on highly adhesive substrata, to which the cells attach as individual units, and grown in vitro for 2 weeks in serum-free, chemically defined media . The behavior of neurons and photoreceptors in vitro was investigated with several techniques; essentially no differences were found between rdle/rdle and rdle/++ cells . Three distinctive cell types could be recognized in cultures of both genotypes towards the end of the first week in vitro: process-free cells, multipolar neurons and rod photoreceptors . There were similarities between rdle/rdle and rdle/++ cultures in the number and morphology of photoreceptor cells, to include the presence of a cilium and a short neurite terminating in a spherule-like body . Moreover, in cultures of both genotypes, only photoreceptors showed opsin immunoreactivity and the antigen recognized by the rod-specific monoclonal antibody RET-P1 . Biochemical and autoradiographic studies demonstrated that rdle/rdle and rdle/++ cells also showed similar uptakes of the putative amino acid neurotransmitters glutamate and aspartate (associated with most of the photoreceptors and only some neurons), and gamma-aminobutyric acid (associated with neurons but absent in photoreceptors) . Thus, according to several parameters, the properties shown by photoreceptor cells were similar in rdle/rdle and rdle/++ cultures during the first week in vitro . Massive photoreceptor cell death was observed in both genotypes during the second week in vitro, coinciding with the time when photoreceptor degeneration occurs in vivo in rd/rd, but not in rd/+ retinas . Photoreceptor death in culture appeared to be specific, since approx . 80% of the non-photoreceptor neurons survived normally during the period when photoreceptor degeneration took place . Several reports from the literature suggest that the period around postnatal days 8-10 represents a critical stage for rd/rd photoreceptors, since they survive until this time but degenerate thereafter . Genetically normal photoreceptors apparently undergo a comparable crisis during maintenance in primary culture, suggesting the involvement of cell-cell contacts and/or retina-derived environmental signals in the survival or rod visual cells. Toxicol Appl Pharmacol, 1988 Aug, 95(1), 61 - 71 Allylamine-induced vascular toxicity in vitro: prevention by semicarbazide-sensitive amine oxidase inhibitors; Ramos K et al.; The present studies were designed to evaluate the role that metabolic activation plays in allylamine (AAM)-induced vascular toxicity . The effects of AAM were evaluated in primary cultures of rat vascular endothelial (VEC) and smooth muscle cells (SMC) . Semicarbazide (SC) and diethyldithiocarbamate (DDC) were used as inhibitors of semicarbazide-sensitive amine oxidase (SSAO) . Clorgyline and pargyline were used as inhibitors of monoamine oxidase (MAO) A and B, respectively . The effect of catalase, a hydrogen peroxide scavenger, on AAM-induced cytotoxicity was also evaluated . Lactate dehydrogenase (LDH) release and morphological alterations were chosen as indicators of cytotoxicity . Confluent cultures of VEC and SMC were exposed to various concentrations of AAM (2-200 microM) in the absence and presence of serum for 4, 12, or 24 hr . High concentrations of AAM (200 microM) alone produced a time-dependent increase in LDH release and morphologic alterations in cultures of both cell types . Lower concentrations of AAM did not compromise the structural integrity of the cells . Semicarbazide (200 microM) or DDC (2 mM), but not clorgyline (10 microM) or pargyline (10 microM), prevented the toxicity of AAM (200 microM) . Allylamine-induced cytotoxicity was partially prevented by catalase (2500 U/ml) . The presence of fetal bovine serum in the medium was not essential for the manifestation of cytotoxicity . Single cell suspensions of VEC or SMC formed acrolein (ACR) when incubated in the presence of AAM . The formation of ACR mediated by SMC was inhibited by SC (20 microM), but not clorgyline (10 microM) . These results support the concept that AAM is oxidatively deaminated by an SSAO present in vascular cells to generate toxic metabolic by-products capable of causing extensive cellular injury. In Vitro Cell Dev Biol, 1988 Aug, 24(8), 771 - 7 Beta-adrenergic receptor characteristics of postnatal rat myocardial cell preparations; Welder AA et al.; Primary myocardial cell cultures and freshly isolated cardiac cells in suspension represent two isolated, whole cell models for investigating cellular transsarcolemmal 45Ca++ exchange in response to a receptor-coupled stimulus . Studies were performed to characterize beta-adrenergic receptor binding, beta-adrenergic receptor mediated cellular calcium (45Ca++) exchange, and viability in purified primary myocardial cell cultures and freshly isolated cardiac cells in suspension obtained from 3- to 5-d-old Sprague-Dawley rats . In addition, beta-adrenergic receptor binding was characterized in whole-heart crude membrane preparations . All three preparations had saturable beta-adrenergic binding sites with the antagonist {125I}iodopindolol {( 125I}IPIN) . The suspensions had a significantly lower Bmax (42 +/- 6 fmol/mg protein) than the membranes and cultures (77 +/- 8 and 95 +/- 10 fmol/mg protein, respectively) . The KD of the cultures (218 +/- 2.0 pM) was significantly higher than that for the suspensions (107 +/- 1.3 pM) and membranes (93 +/- 1.3 pM) . Viability was significantly lower in the suspensions (57%) when compared to 94% viability in myocardial cell cultures after 3 h of incubation in Kreb's Henseleit buffer . Incubation of the cultures with 5.0 X 10(-7) M isoproterenol resulted in a significant increase in 45Ca++ exchange as early as 15 s . In contrast, 45Ca++ exchange into the suspensions was not increased . Although both primary cell cultures and cardiac cells in suspension possess saturable beta-adrenergic receptors, only the monolayer cultures exhibited functional beta-adrenergic receptor-mediated 45Ca++ exchange . Of the two intact cell models investigated, these data suggest that primary myocardial cell cultures are more suitable than cell suspensions for investigating beta-adrenergic receptor binding and functions in the postnatal rat heart. Mol Immunol, 1988 Aug, 25(8), 713 - 8 Cross-reaction of a monoclonal antibody to human MHC class II molecules with rabbit B cells; Dubiski S et al.; Monoclonal antibodies, 21w4 and 44H10, against human MHC class II determinants, were analysed for their reactivity with rabbit lymphoid cells . Both MAb bind to all human B lymphoblastoid lines, irrespective of their HLA-DR phenotype . HLA-DR antigens, purified by affinity to 21W4 IgG Sepharose, can be precipitated with 44H10 MAb, indicating that both antibodies react with the same molecules . Competitive inhibition studies with purified MAb IgG show that 44H10 and 21w4 recognize different epitopes of HLA-DR molecules . The 21w4 MAb, previously shown to cross-react with cells of pig, mouse and sheep, does not react with rabbit lymphoid cells . The 44H10 MAb binds to lymphoid cell suspensions prepared from rabbit appendix, spleen and mesenteric lymph nodes . Its reactivity correlates with that of RABELA, a polyclonal antibody specific for rabbit B cells . Mesenteric lymph node and spleen cell suspensions, depleted of sIg+ cells, are devoid of 44H10+ cells, while similarly-treated appendix B cells still contain a subset of B cells which are sIg-, RABELA+ and 44H10+ . These studies thus report on the presence of MHC class II determinants on rabbit B cells, cross-reacting with human HLA-DR. Cell Biophys, 1988 Aug, 13(1), 55 - 63 Light-stimulated ultraweak photon reemission of human amnion cells and Wish cells; Scholz W et al.; Photon reemission in the ultraweak intensity range that is observed after irradiation of cell suspensions with light, reveals characteristic differences between normal human amnion cells and transformed Wish cells from the same parental tissue . The reemission kinetics, approximated best by a hyperbolical process, were studied as a function of cell density, showing that: malignant Wish cells have a photon storage capacity that is not improved by increasing the cell density; and that normal amnion cells exhibit a photon storage capacity that strongly increases with increasing cell density . The interpretation of this effect and the nature of the emitter are discussed. FEBS Lett, 1988 Jul 18, 234(2), 349 - 52 NMR water-proton spin-lattice relaxation time of human red blood cells and red blood cell suspensions; Sullivan SG et al.; NMR water-proton spin-lattice relaxation times were studied as probes of water structure in human red blood cells and red blood cell suspensions . Normal saline had a relaxation time of about 3000 ms while packed red blood cells had a relaxation time of about 500 ms . The relaxation time of a red cell suspension at 50% hematocrit was about 750 ms showing that surface charges and polar groups of the red cell membrane effectively structure extracellular water . Incubation of red cells in hypotonic saline increases relaxation time whereas hypertonic saline decreases relaxation time . Relaxation times varied independently of mean corpuscular volume and mean corpuscular hemoglobin concentration in a sample population . Studies with lysates and resealed membrane ghosts show that hemoglobin is very effective in lowering water-proton relaxation time whereas resealed membrane ghosts in the absence of hemoglobin are less effective than intact red cells. Cancer Res, 1988 Jul 15, 48(14), 4065 - 72 Morphological study of the interaction of intravascular tumor cells with endothelial cells and subendothelial matrix; Crissman JD et al.; Multiple steps or events have been described as essential in the metastatic cascade . Tail vein injection of single cell suspensions was used to study the ultrastructural details of the events involved in the initial arrest and attachment of circulating tumor cells . Lewis Lung Carcinoma (3LL) and a mammary adenocarcinoma (16c) were compared to a previous ultrastructural study of B16 amelanotic melanoma (B16a) detailing morphological events in the initial arrest and attachment of tumor cells in lung . The three murine tumors followed similar steps and varied only slightly in the time sequence of the steps . We observed the following steps: (a) initial arrest of tumor cells was characterized by an intimate tumor endothelial cell contact; (b) platelet activation and aggregation was noted by two minutes . Platelet aggregation continued for 1-4 h until a thrombus formed; (c) after approximately 4 h endothelial cell separation with extension of the tumor cell to the subendothelial matrix was noted; (d) at approximately 24 h the tumor cell associated thrombus dissipated and the attached tumor cells were exposed to a reestablished circulation . (e) mitoses were observed after 24 h with cell division and the development of intravascular tumor nodules; (f) the final step in the extravasation sequence was dissolution of the basement membrane by the attached tumor cells. Eur J Biochem, 1988 Jul 15, 175(1), 187 - 91 Activation of inositol phosphate formation by circulating noradrenaline but not by sympathetic nerve stimulation with a similar increase of glucose release in perfused rat liver; Puschel GP et al.; In the isolated rat liver perfused in situ, stimulation of the nerve bundles around the hepatic artery and portal vein caused an increase of glucose and lactate output and a reduction of perfusion flow . These changes could be inhibited completely by alpha-receptor blockers . The possible involvement of inositol phosphates in the intracellular signal transmission was studied . 1 . In cell-suspension experiments, which were performed as a positive control, noradrenaline caused an increase in glucose output and, in the presence of 10 mM LiCl, a dose-dependent and time-dependent increase of inositol mono, bis and trisphosphate . 2 . In the perfused rat liver 1 microM noradrenaline caused an increase of glucose and lactate output and in the presence of 10 mM LiCl a time-dependent increase of inositol mono, bis and trisphosphate that was comparable to that observed in cell suspensions . 3 . In the perfused rat liver stimulation of the nerve bundles around the portal vein and hepatic artery caused a similar increase in glucose and lactate output to that produced by noradrenaline, but in the presence of 10 mM LiCl there was a smaller increase of inositol monophosphate and no increase of inositol bis and trisphosphate . These findings are in line with the proposal that circulating noradrenaline reaches every hepatocyte, causing a clear overall increase of inositol phosphate formation and thus calcium release from the endoplasmic reticulum, while the hepatic nerves reach only a few cells causing there a small local change of inositol phosphate metabolism and thence a propagation of the signal via gap junctions. Biochim Biophys Acta, 1988 Jul 7, 942(1), 96 - 106 The influence of tetraphenylborates (hydrophobic anions) on yeast cell electro-rotation; Arnold WM et al.; The action of a series of tetraphenylborate ion (TPB) derivatives on yeast cells was studied by electro-rotation of the pre-treated cells . TPB derivatives in which all four phenyl groups were substituted with fluorine, chlorine or trifluoromethyl were much more toxic than the unsubstituted compound, the effect increasing dramatically with increasing size of substituents . These observations suggest that the toxicity of these hydrophobic ions is determined mainly by their size and possibly also by the chemical inductivity of their substituent groups . The order of the toxicities of these ions was in fair agreement with literature values for their translocation rates across artificial bilayers . Incubation times of 3 h were used as standard, longer incubations (up to 48 h) showed that the number of cells affected by low doses of TPB increased with the logarithm of time after the first hour of incubation . Although measurements of the percentage of cells showing co-field rotation showed that controls were not adversely affected by incubations as long as 9 h, rotation spectra showed that some cells suffer loss of internal conductivity during extended incubations . Decrease of the pH of the incubation medium, or inclusion of high concentrations of NaCl or KCl, potentiated the effects of these hydrophobic ions . The toxicity developed slowly, and the sensitivity of the assay was only very weakly dependent on the cell suspension density. J Reprod Fertil, 1988 Jul, 83(2), 647 - 53 Effect of A-nor steroids and oestradiol on progesterone production by human luteal cells; Wang HZ et al.; Cell suspensions were prepared from human corpora lutea obtained during the mid-luteal phase . Progesterone production was assessed after short-term incubation of luteal cell suspensions . Luteal cells were very sensitive to hCG, the concentration required for 50% maximum response being 0.01 i.u./ml, and the response was 5 times higher than the basal production . Oestradiol (1-100 microM) induced a significant dose-related decrease in both basal and hCG-stimulated progesterone production . The A-nor steroidal compounds anordrin and AF-45 reduced hCG-stimulated progesterone production only at the high concentration of 100 microM . The ED50 values were approximately 3 microM, 75 microM and 100 microM for oestradiol, AF-45 and anordrin respectively . Anordrin showed no significant effects on basal progesterone production . In addition, oestradiol markedly inhibited the activity of 3 beta-hydroxysteroid dehydrogenase in luteal cells, expressed by the conversion of pregnenolone to progesterone, but the inhibitory effects of anordrin and AF-45 were negligible or relatively low . The effects of anordrin and AF-45 were different from those of oestradiol on progesterone production by human luteal cells in vitro, indicating that neither substance is likely to be a useful luteolytic agent in women. Brain Res, 1988 Jul 1, 470(1), 146 - 50 Early appearance of type II astrocytes in developing human fetal brain; Elder GA et al.; To compare rat and human glial development, we examined the cellular composition of human fetal brain in short-term cultures and fresh cell suspensions from fetal ages ranging from 7 to 16 weeks, utilizing the cell type-specific markers which define glial subsets in rats . Here we report that unlike the early rat central nervous system (CNS), 7-10 week human fetal brain contains mostly astrocytes that can be characterized as type II rather than type I . Type I astrocytes become more prevalent in 16-week gestational age human brain . Although cells morphologically and immunocytochemically similar to the rat 02-A cell are found in human fetal brain and spinal cord, these cells were not induced to express galactocerebroside in serum-free media and did not have vimentin-containing intermediate filaments as do rat 02-A cells . These results suggest that functional differences may exist between rat type I and type II astrocytes and phenotypically similar cells found in humans. Hematol Oncol, 1988 Jul-Sep, 6(3), 205 - 11 Rearranging antigen-receptor genes in enriched Reed-Sternberg cell fractions of Hodgkin's disease; Cossman J et al.; Molecular genetic analysis of rearranging antigen-receptor genes in non-Hodgkin's lymphomas has revealed the clonality and lineage in the majority of cases . In an analogous approach, we sought to apply gene rearrangement analysis to Hodgkin's disease to understand better the clonality and origin of this disorder . However, the putative neoplastic cell of Hodgkin's disease, the Reed-Sternberg cell and its variants, is extraordinarily rare in most cases of Hodgkin's disease . On the average, Reed-Sternberg cells and variants represent 0.1 per cent of total cell suspensions of nodular sclerosing Hodgkin's disease . As this frequency is below the minimum threshold of sensitivity of the Southern blot assay, we attempted to enrich for Reed-Sternberg cells before DNA extraction and analysis . Using either elutrition or Percoll density gradient centrifugation, we were able to enrich the percentage of Reed-Sternberg cells and variants to above 1 per cent in five cases of nodular sclerosing Hodgkin's disease . In three of these cases, immunoglobulin gene rearrangements were identified, but no T cell receptor gene rearrangements were seen . No rearrangements were detected in unseparated cells or in the Reed-Sternberg cell-depleted fractions . In addition, the L428 Hodgkin's disease cell line was found to have one rearranged and one deleted heavy-chain gene, a rearranged kappa gene, a rearranged lambda gene, and a single rearranged beta allele . No rearrangements of the T gamma gene were found in L428 . Taken together, these findings indicate that clonal cell populations are present in Hodgkin's disease and suggest the possibility of a clonally expanded lymphoid cell in this disorder. J Neurosurg, 1988 Jul, 69(1), 121 - 6 Brain xenografts: the effect of cyclosporin A on graft survival; Howard MA 3rd et al.; Animal models of Parkinson's disease and Alzheimer's disease have shown dramatic functional improvement after transplantation of embryonic neurons into denervated regions of the adult brain . Because of the ethical and logistic problems associated with the use of human embryonic brain tissue, cross-species transplants are an attractive alternative . An experimental model of cross-species brain transplantation was developed to evaluate cell survival in untreated and cyclosporin A (CyA)-treated animals . Cholinergic ventral neurons from embryonic mice were transplanted into the frontal lobes of 18 adult Sprague-Dawley rats using a cell suspension technique . Nine animals were treated for 13 days with CyA (10 mg/kg/day) and nine were not treated . Twelve weeks after transplantation, frozen sections through the transplant volume were obtained . Alternate sections were prepared with hematoxylin and eosin and acetylcholine esterase stains . Cell counts through a 2-cu mm volume incorporating the transplant were compared to a contralateral control volume . Eight of the nine untreated transplants were successful (mean transplant cells +/- standard error of the mean: 90.7 +/- 19.4/2 cu mm) . All of the nine CyA-treated transplants survived, with mean transplant count 28.7 cells/2 cu mm greater than untreated transplants (mean increase 28.7: p less than or equal to 0.05, Wilcoxon matched-pairs signed ranks test) . It is concluded that: 1) this model is useful for quantitating transplant cell survival; 2) untreated xenografts survive well; and 3) a 13-day course of CyA improved long-term graft survival. J Immunol, 1988 Jul 1, 141(1), 315 - 23 Age-related variation in the proportion and activity of murine liver natural killer cells and their cytotoxicity against regenerating hepatocytes; Itoh H et al.; We investigated the distribution of liver NK cells in mice of various ages and their cytotoxicity against regenerating hepatocytes . Liver NK cells were identified by asialo GM1 antibody in mononuclear cell suspension from the liver, whereas NK activity was assayed against YAC-1 target cells . Mononuclear cells in the liver consisted of more than 25% NK cells with potent NK activity in C3H/He mice, 8 wk of age . The strain-specific distribution (C3H/He greater than C57BL/6 greater than DBA/2) of liver NK cells was the same as those in the spleen and blood . The proportion of liver NK cells and the level of NK activity in C3H/He mice were further demonstrated to vary depending on age, in that both the proportion and the function were generated at 4 wk of age, reached a maximum between the 6th and 8th wk, and then rapidly decreased around the 9th wk . The appearance of an increased number of NK cells in the liver seemed to coincide with the slowing of the rapid increase of murine liver weight . We then investigated whether liver NK cells mediated their cytotoxicity against regenerating hepatocytes . Both specific 51Cr-release assay and single cell cytotoxicity assay demonstrated that liver NK cells were significantly cytotoxic against regenerating hepatocytes in partially hepatectomized liver, but to a lesser extent against normal hepatocytes in resting liver . Morphologic study revealed that normal liver predominantly consisted of hepatocytes with binuclei (greater than 60%) but that regenerating liver mainly consisted of hepatocytes with a single nucleus (greater than 70%) . One-nucleus hepatocytes were more susceptible to the cytotoxicity of liver NK cells . A comparative study of restoration kinetics of the liver weight and the number of liver NK cells after partial hepatectomy also showed a unique relationship . These results raise the possibility that liver NK cells might be responsible for regulating hepatocyte growth. Endocrinology, 1988 Jul, 123(1), 113 - 9 Altered differentiated cell surface properties of transformed (RINm5F) compared with native adult rat pancreatic B cells; Halban PA et al.; RINm5F cells, a line derived from a rat insulinoma, are frequently used as a model for studying pancreatic B cell structure and function . These transformed cells are known, however, to be different from native B cells in a number of biochemical respects . We have now compared the surface features of RIN cells and native B cells in two different ways: 1) Dispersed cells from islet obtained from adult rats can reassociate spontaneously in culture to form aggregates (pseudoislets) with cellular organization similar to that of intact native islets (a central B cell core surrounded by a discontinuous mantle of non-B cells) . Native islet cells and RIN cells were mixed together and allowed to reaggregate . Examination by immunocytochemistry and transmission electron microscopy showed that the aggregates contained all cell types present in the original mixed cell suspension (native B- and non-B cells, and RIN cells) . The native B cells were centrally located, surrounded by zones of non-B cells, as in islets . The RIN cells, however, were restricted to the periphery and as such not recognized as native B cells . 2) R2D6, a monoclonal antibody, binds selectively to a ganglioside on the surface of islet B, but not non-B, cells . The role of this ganglioside is not known . RIN cells were incubated with R2D6 followed by a fluorescently labeled second antibody . Analysis by flow cytofluorometry indicated that the monoclonal antibody had bound (stained) only 3-15% of the RIN cells . These R2D6 positive cells were sorted from R2D6 negative cells and subsequently shown to have a lower DNA content . Expression of the R2D6 target ganglioside on RIN cells thus appears to be cell cycle dependent . Based on two different criteria, RINm5F cells do not therefore share surface features in common with native B cells . The cell cycle dependent expression of a B cell surface antigen by the RIN cells might, however, be a useful model for studying the regulation of ganglioside turnover. Rev Argent Microbiol, 1988 Jul-Sep, 20(3), 107 - 18 {Effect of quinones and nitrofurans on Trypanosoma mega and Crithidia fasciculata}; de Pahn EM et al.; Demonstration of trypanocidal effects in vitro is a first step for the development of new antichagasic drugs . In order to obtain an experimental model allowing the pre-screening of potential trypanocides for Trypanosoma cruzi in a short time and under safe conditions, the trypanosomatids T . mega and C . fasciculata were assayed for their response to a) compounds known for their action on T . cruzi, and b) compounds not tested before on the latter . The drugs were assayed on the organisms growth in a liquid culture medium, cell multiplication being measured by the medium turbidity increase, using a photoelectric colorimeter previously calibrated with cell suspensions of known concentration . A series of quinones (Lapachones and related compounds), naftoquinone-imines, benzoquinones (perezone and dihydroperezone), a quinol (miconidine) and several nitrofurans, including nifurtimox and (5-nitro-2-furfurylidene)-amino (NF-group) derivatives, inhibited the flagellates growth, specially T . mega, with half-maximal inhibitory concentrations lesser than 5.0 microM, for the most active compounds . T . mega response to nifurtimox, NF-derivatives and beta-lapachone was in close agreement with that of T . cruzi . Cultures of T . mega in the presence of NF-pyrazole, NF-indazoles and NF-imidazole but not nifurtimox, showed irreversible damage since, after re-incubation in fresh medium without inhibitor, these cells grew significantly less than their corresponding controls . Similar effects were observed in C . fasciculata, with beta-lapachone and one naftoquinone-imine . Our results qualify T . mega as an adequate experimental model for the assay of antichagasic agents, as C . fasciculata and T . brucei brucei do for the african trypanosomes. No To Shinkei, 1988 Jul, 40(7), 623 - 8 {Transplant-induced recovery from 6-OHDA lesions of the nigro-striatal dopamine neurons in mice}; Shimizu K et al.; Attempts to reconstruct the damaged nigrostriatal pathway in experimental models of Parkinson disease have thus far been carried out in animals with neurotoxically induced dopamine deficiency . Our study established that unilateral 6-hydroxydopamine (6-OHDA) lesions of the nigrostriatal-dopamine (DA) neurons produced a well-characterized functional asymmetry in the behavior of C57BL/6 (H-2b) mice . The intraperitoneal administration of methamphetamine induced ipsilateral rotation at 7-20 turns/min . 11 x 10(6) syngenic DA-rich cells of embryonic ventral mesencephalon were stereotaxically transplanted in the caudate-putamen . A complete recovery of methamphetamine-induced rotational response was produced around the 60-th day after the syngenic cell suspension graft . And a complete compensation of the rotational response was also brought about with the DA-rich cells from embryonic ventral mesencephalon (crown-rump length; 10-13 mm) of allogenic C 3 H/HeN (H-2k) mice . The FACS IV analysis revealed no H-2 (Kk and Iak) antigens before transplantation of these embryonic cells . Immunohistochemistry showed that the dopaminergic fibers had grown predominantly into the ipsilateral caudate-putamen . These results provide evidence of integration of syngenic and allogenic grafts and host tissue . And the immunological response in the transplanted brain are under investigation. Z Naturforsch {C}, 1988 Jul-Aug, 43(7-8), 479 - 84 Novel glucoalkaloids from Rauwolfia cell cultures--acetylrauglucine and related glucosides; Ruyter CM et al.; From cell suspension cultures of Rauwolfia serpentina grown in an optimized production medium for the glucoalkaloid raucaffricine, a novel glucoalkaloid was isolated and identified as 17-O-acetyl-21-O-beta-D-glucopyranosyl-ajmaline (acetylrauglucine) . This alkaloid is formed in very small amounts (less than 5 x 10(-4)%) . The biogenetically related N alpha-demethylated base (acetyl-nor-rauglucine) and the deacetyl product rauglucine have also been detected in culture extracts . In addition 21(R)-(beta-D-glucopyranosyl)-hydroxy-sarpagan-17-al has been isolated and identified as an artifact which originates from raucaffricine. ASAIO Trans, 1988 Jul-Sep, 34(3), 773 - 7 In vitro evaluation of a pyridoxalated hemoglobin polyoxyethylene conjugate in reversing cell sickling; Yabuki A et al.; In sickle cell disease (SCD) microcirculatory blockage by red blood cells (RBC) occurs because of their low oxygen concentration, which results in both sickling and painful crises . Pyridoxalated hemoglobin polyoxyethylene (PHP), developed from human RBC hemoglobin (Hb) by chemical modification as an oxygen carrier, was evaluated in vitro for its ability to reverse cell sickling . PHP solutions of 6 or 8 g % Hb and a P50 of 20 mmHg were evaluated . RBC were obtained from SCD patients treated by exchange transfusion . The in vitro positive pressure filtration method (47 mm 5 microns Nuclepore membrane) was used . Comparisons of PHP with low oxygen carrier solutions (Hespan, saline) were made at flows of 0.43 to 6.0 ml/min . PO2 increases in a 20% mixture of air saturated solutions with deoxygenated sickle cell suspensions (SCS), at a hematocrit of 1%, were significantly higher in PHP as compared with saline and Hespan . The filtration resistance of deoxygenated SCS mixed with PHP was significantly lower than that of deoxygenated SCS mixed with saline and Hespan, and was comparable to that of air oxygenated SCS . By the 20% (v/v) addition of air saturated PHP to deoxygenated SCS, 89 +/- 2% of the sickled cells were unsickled . A novel artificial capillary system (ACS) modeling the dynamics of the microcirculation of the body was used . With the ACS plugged with deoxygenated cells, perfusion with oxygenated cell-PHP solutions was significantly more efficient in reversing the blockage than oxygenated saline and Hespan solutions . PHP reverses cell sickling by its effective delivery of oxygen. ASAIO Trans, 1988 Jul-Sep, 34(3), 578 - 80 An in vitro model for human endothelial cell seeding of a small diameter vascular graft; Kent KC et al.; A precise system was devised to measure the kinetics of attachment of human venous endothelium to a variety of materials and substrates . Cells were labelled in a postconfluent state with tritiated thymidine, harvested, and a cell suspension seeded into a 4 mm PTFE graft . After a 90 minute incubation period, one half of the graft segment was sacrificed and the remaining portion placed in a perfusion system (225 cc/min) for 1 hour . Graft segments, effluents, and seeding suspension were assayed in a beta scintillation counter . The percentage of cells that attached pre- and postperfusion were determined, as well as the retrieval of tritium from the system . Initially, 71% of seeded cells attached to grafts coated with fibronectin, with significantly less (60%) remaining attached after perfusion . Only 10% of cells initially attached to uncoated grafts, with 4% retained postperfusion . Retrieval of tritium averaged 102 +/- 10% for all experiments . This system determines both pre- and postperfusion attachment of human endothelial cells to vascular grafts following manipulation of numerous variables, including graft material, substrate, incubation time, and seeding density . An optimal seeding protocol for human trials can thus be determined. ASAIO Trans, 1988 Jul-Sep, 34(3), 375 - 85 Couette membrane filtration with constant shear stress; Fischel RJ et al.; Recent developments in the field of blood component separation have revealed the usefulness of membrane filtration using couette type configurations and Taylor vortices as an efficient and effective method . The authors have analyzed in detail the physical and chemical effects on whole blood separated into protein rich plasma, and concentrated red blood cell suspensions, using this technique . The authors also have calculated and demonstrated the technical specifications required to provide laminar flow with Taylor Vortex formation throughout the device, as well as those required to retain constant shear stress on the blood components as viscosity changes . By maintaining constant shear stress below a critical level, it is possible to avoid shear induced hemolysis and to maintain maximal separation efficiency throughout the procedure . The device has further been designed to alter the filtration velocity along the membrane so that the critical filtration velocity is nowhere exceeded, i.e., concentration polarization effects are prevented. Agents Actions, 1988 Jul, 24(3-4), 261 - 5 Enhancement by cimetidine of chemotactic peptide-stimulated ATP release and chemiluminescence in human neutrophils; Tarnok I et al.; As determined by luciferase-luciferin, we recently found that the H2-blocker CIM considerably increased the ATP release from fMLP-stimulated PMN . This observation correlates well with our previous {1} regarding the enhancement of superoxide output (chemiluminescence) in in human neutrophils . by CIM plus fMLP . In order to compare the ATP release from PMN of different donors, a standard procedure has been developed consisting of the determination of the ATP present initially in the cell suspension (without stimulation), ATP release after stimulation with fMLP, and ATP release in the presence of CIM plus fMLP . The whole ATP content per neutrophil was determined after ultrasonication of the cells as well . The mean value of the initially present ATP was 0.45 x 10(-17) mol/cell in the suspension . Stimulation with fMLP plus CIM yielded within 5-10 minutes considerably higher ATP amounts than fMLP alone . The corresponding and statistically significantly different mean values were 2.46 x 10(-17) mol/PMN (s.d . = 1.047) and 1.38 x 10(-17) mol/PMN (s.d . = 0.55), respectively . The whole ATP per neutrophil was found to be 1.22 x 10(-15) mol (mean; s.d . = 0.60) and thus, the stimulation with CIM plus fMLP released about 2.0 per cent, with fMLP alone about 1.0 per cent of the whole ATP . CIM without fMLP did not enhanced the ATP release during the reaction time applied . On the other hand, fMLP-stimulated, lucigenin-amplified chemiluminescence determinations were carried out in the presence of CIM as well; contrarily to our previous method, CIM was dissolved in PBS without DMSO, because DMSO inhibited the chemiluminescence slightly.(ABSTRACT TRUNCATED AT 250 WORDS) Immunobiology, 1988 Jul, 177(3), 293 - 304 Lack of correlation between in vivo peroxidatic activity patterns and tumoricidal activity in vitro of peritoneal macrophages after intraperitoneal immunization with tumor cells; Dullens HF et al.; Immunization of C57BL mice with one inoculum of 10(7) DBA/2-derived SL2 lymphosarcoma cells resulted in a +/- 20-fold increase in the total number of peritoneal cells . The number of macrophages showed a 10-fold increase from 3 x 10(6) (control mice) to 3.4 x 10(7) cells at day 8 after immunization . Within this macrophage population, four different cell types, based on the ultrastructural peroxidatic activity patterns, could be distinguished: exudate macrophages, resident macrophages, resident-exudate macrophages and peroxidatic-activity-negative macrophages . The number of exudate macrophages significantly increased in the peritoneal cavity after immunization: at day 8 after immunization, a peak value of 10(7) cells was observed . At the same time, there were 2.2 x 10(7) peroxidase-activity-negative macrophages present (representing the control value x 50) . Significant in vitro tumoricidal activity of the isolated macrophages could not be measured until 8 days after immunization . At that time, a cytotoxicity index of 68 was reached . After immunization of the C57BL mice with 3 injections with allogeneic SL2 cells, there were no dramatic changes in the number of peritoneal cells after the last immunization . Only immediately after the last immunization was a minor increase in peroxidatic-activity-negative macrophages seen . But already at 5 days after the last immunization, the composition of the peritoneal suspension was similar to that of non-immunized mice with predominantly resident macrophages . The cytotoxicity of the peritoneal macrophages from hyperimmunized mice was constantly high during 1-15 days after the last immunization (cytotoxicity index ranged from 66-72) . In order to study which type(s) of macrophage(s) (resident, exudate, resident-exudate or peroxidatic-activity-negative) is/are responsible for the cytotoxicity measured in vitro, peritoneal cell suspensions (obtained after immunization) were fractionated according to their affinity to wheat germ agglutinin (WGA) coupled to Sepharose columns . Comparison of the values of cytotoxicity measured before and after separation into "subtypes" of the macrophages revealed that the expression of cytotoxicity is not correlated with any of the "sub-types", especially when the peroxidatic activity pattern is is taken as a criterion. Ann Inst Pasteur Immunol, 1988 Jul-Aug, 139(4), 361 - 81 Mechanism of K562-induced human natural killer cell inactivation using highly enriched effector cells isolated via a new single-step sheep erythrocyte rosette assay; Abrams SI et al.; In this study, we used preparations highly enriched in human natural killer (NK) cells to further characterize the mechanism of target-cell-induced NK inactivation . Highly enriched populations of NK cells were obtained by a newly developed, single-step sheep red blood cell rosette assay . This method, which did not require any incubation steps to facilitate cell contact, permitted a rapid and efficient isolation of NK cells from adherent-cell-depleted peripheral blood lymphocytes . The non-rosetted cells had high NK activity, possessed large granular lymphocyte (LGL) morphology and expressed the NK-associated antigens Leu-11a, Leu-7, OKM1 and NKH-1 . In contrast, the rosetted cells had significantly lower NK activity, possessed typical lymphocyte morphology and expressed the T-cell-associated marker OKT3 . Next, we examined the ability of these NK-enriched effector cells (ECc) to become inactivated by K562 . Functional studies revealed that ECc lost greater than or equal to 95% of their lytic capacity following incubation with K562 at a ratio of 2/1 for 6 h . However, to achieve this level of inactivation, it was essential that the cell suspension be gently mixed every 90-120 min . Inactivation was not due to cell death and did not reflect changes in the percentages of cells bearing the Leu-11a, Leu-7, OKM1 and NKH-1 antigens, but was associated with an increase in cell surface concentration of OKM1 . As judged by gross morphology, the percentages of LGL in ECc before and after treatment with K562 were essentially the same . Finally, K562-treated ECc also lost their ability to mediate antibody-dependent cellular cytotoxicity (ADCC), suggesting that both NK-cell-mediated cytotoxicity and ADCC may involve a common lytic pathway. Blood, 1988 Jul, 72(1), 282 - 6 Bivariate flow karyotyping in human Philadelphia-positive chronic myelocytic leukemia; Arkesteijn GJ et al.; Chromosome analysis on clinical leukemia material was done by means of flow cytometry (flow karyotyping) to investigate the applicability of this technique in the detection of leukemia-associated abnormalities . Flow karyotyping was performed on blood or bone marrow samples from eight patients with chronic myelocytic leukemia (CML) after a culture period of four days and arresting the cells in metaphase during the last 16 hours . Discontinuous density gradient centrifugation proved to be essential in removing debris and dead cells from the cell suspensions . By this procedure the mitotic index increase ranged from 2 to 80 times initial values . Chromosomes were isolated and stained with two base pair-specific fluorochromes, ie, chromomycin A3 and Hoechst 33258, and run through a specially designed dual-laser beam flow cytometer . Generally, 20,000 chromosomes or more were measured . The data were computer stored in list mode . Besides the clear detection of the specific Philadelphia chromosome, trisomies and other additional chromosomal aberrations {like an i(17q)} were visualized . Quantitative analysis revealed the percentage of subclones containing a certain chromosomal anomaly . Conventional cytogenetic analysis confirmed these findings . In seven of eight cases, CML could be diagnosed on the basis of the presence of a Philadelphia chromosome in the flow karyogram . In one of these seven, the conventional cytogenetic analysis was unknown at that time . The remaining six all matched the standard cytogenetics . The one failure out of eight could be attributed to the specific stimulating conditions in the culture . Although it is impossible by this technique to determine the position of the breakpoint, the involved chromosomes in the translocation event could be identified . In some cases, low percentages of aberrations could not be detected . This study shows that CML can be diagnosed on the basis of flow karyotypic results . Additional chromosomal aberrations can be detected provided that changes in the amount of DNA per chromosome have occurred . Exact quantification of the composition of subclones in the case of mosaicism appear difficult. J Comp Neurol, 1988 Jul 1, 273(1), 26 - 41 Development of intracerebral dopaminergic grafts: a combined immunohistochemical and autoradiographic study of its time course and environmental influences; Abrous N et al.; The aim of the study was to obtain a description of some aspects of the development of intracerebral dopaminergic grafts, namely, the time course of the glial reaction and its relation to cell division on one hand, and the development of graft-originated innervation and its dependence on adequate matching of the implanted neurons and target site on the other hand . Cell suspensions obtained from the mesencephalon or hypothalamus of embryonic day (ED) 14 rat embryos were implanted into the striatum or lateral hypothalamus of adult rats following the destruction of the nigrostriatal system of the hosts . Animals were sacrificed at different postimplantation times, and the development of the graft was followed by immunohistochemistry by using antisera directed against tyrosine hydroxylase (TH) or glial fibrillary acidic protein (GFA) . Furthermore, the existence of cell division at various times following implantation was examined by performing autoradiography on immunostained sections after prior intraventricular administration of 3H-thymidine to the host . The first stage of the development of intracerebral grafts was characterized by the existence of intense cell division within the grafted tissue, lasting about 2 weeks, and also in the host tissue surrounding the graft, lasting only about 6 days . The cell division in the host tissue was paralleled by the existence of a strong glial reaction which, however, did not extend into the graft itself . Glial reaction in the host tissue gradually decreased at later times and disappeared by 4 weeks postimplantation without leaving behind a noticeable glial scar . The graft itself was, however, transiently filled with a population of reactive astroglial cells between 3 and 6 weeks postimplantation . Within grafts of mesencephalic tissue located in the striatum TH-positive neurons were distributed evenly at short times postimplantation (2-6 days) . At later time a compartmentation could be observed, with TH-positive neurons being aligned along the graft-host interface or clustered within the graft itself . Innervation of the host tissue by TH-positive fibers increased between 1 and 6 weeks postimplantation . On the other hand, no compartmentation and reinnervation of surrounding host tissue was observed for intrahypothalamic grafts of mesencephalic tissue or intrastriatal grafts of hypothalamic tissue . This last observation indicates that adequate matching of implanted neurons and target tissue plays an important role in the development of intracerebral dopaminergic grafts. Am J Physiol, 1988 Jul, 255(1 Pt 2), F22 - 32 Asymmetry of plasma membrane lipid order in Madin-Darby Canine Kidney cells; Le Grimellec C et al.; Fluorescence anisotropy experiments have been done to estimate, in situ, the lipid order of the plasma membrane of polarized Madin-Darby Canine Kidney cells (MDCK) grown on glass cover slips and labeled by 1-{4-(trimethylamino)phenyl}-6-phenylhexa-1,3,5-triene (TMA-DPH), a specific marker of the plasma membrane of living cells . Fluorescence microscopy, back-exchange, and quenching experiments indicated that TMA-DPH labeled the highly ordered (r greater than or equal to 0.32, 37 degrees C) apical domain of the plasma membrane of confluent monolayers . Opening of tight junctions or addition of the probe to cell suspensions resulted in a homogeneous distribution of TMA-DPH over the cell surface and in a marked decrease in anisotropy (0.27 less than or equal to r less than or equal to 0.29) that was due neither to a direct effect of Ca2+ on the probe nor to a change in fluorescence lifetime . Our data indicate that the apical domain, likely the external leaflet, of the plasma membrane of polarized MDCK cells is much more ordered than its basolateral counterpart. Tsitologiia, 1988 Jul, 30(7), 903 - 7 {The possible use of PKZh-type devices (liquid purity control devices) in the analysis of cell suspensions}; Ivanov NR et al.; PKZh means "device for liquid purity control" . A possibility is considered to use the native PKZh type device for carrying out quantitative analyses of cellular suspension components, for routine bacterial suspension, agglutinated bacterial suspension and erythrocyte suspension . The flowing photometric principle of particle recording, used in the device, allows to analyse biological suspensions with small amounts of components . The device provides a differential count of some cells and their conglomerates in six dimensional ranges, within the frames of 1-25 micron or higher . The time consumption for one sample analysis is 10-15 seconds. Cytometry, 1988 Jul, 9(4), 303 - 8 Identification of cell subpopulations by dual-color surface immunofluorescence using biotinylated and unlabeled monoclonal antibodies; Cohen JH et al.; A new method of dual-color immunofluorescence is presented for analysis of surface antigen distribution among heterogeneous cell suspensions . It involves flow cytometric analysis of cells stained with a biotinylated first monoclonal antibody and/or with an unlabeled second monoclonal antibody . After addition of streptavidin-phycoerythrin and/or fluoresceinated goat antimouse immunoglobulin antibody, single-cell fluorescence intensities are measured and biparametric graphic representations are obtained, allowing one to determine the percentage of cells stained by each of the monoclonal antibodies or both . The validity of the method was assessed on human peripheral blood mononuclear cells by using three sets of two monoclonal antibodies: CD8 and CD5, CD3 and CD4, CD11 and HLA-DR . The results showed that dual staining did not induce significant quenching or competition between pairs of antibodies . The procedure is simple and sensitive . It requires only minute amounts of monoclonal antibodies . It is readily applicable to the screening of hybridoma supernatants and to the characterization of new antibodies to cell surface antigens with respect to well-defined markers. Clin Exp Metastasis, 1988 Jul-Aug, 6(4), 325 - 32 Correlation between tumour antigens and malignancy in BKV-transformed hamster cells; Rosciani C et al.; Hamster kidney cells transformed by BK virus (HKBK cells) were studied in order to correlate the membrane tumour antigens to the metastatic capability . The presence of the tumour associated surface antigen (TASA) on the surface of HKBK cells was detected by the immunofluorescence test on live cell suspensions . The metastatic ability was investigated by inoculating HKBK cells subcutaneously (s.c.) into newborn hamsters, s.c., intraperitoneally (i.p.) and in the foot pads into adult hamsters, and s.c . into adult hamsters previously immunized with surface antigens extracted from HKBK cells . The results indicate that there is a correlation between the appearance of tumour antigens on the cell surface and the metastatic ability: HKBK cells at low passage (about 30 subcultures after transformation) showed the capping of TASA in the cell membrane and low metastatic ability, whereas HKBK cells at high passage (about 130 subcultures after transformation) exhibited a diffuse appearance of TASA in the cell surface and were highly metastatic. Immunol Lett, 1988 Jul, 18(3), 219 - 23 Preparation of both DNA and RNA for hybridization analysis from limiting quantities of lymphoid cells; Pearse MJ et al.; This report describes a method for preparing both DNA and RNA simultaneously from as few as 5 X 10(5) lymphoid cells . The method is suitable for cultured cells or any tissue from which a cell suspension can be prepared and for the small samples of purified cells obtained by fluorescence activated cell sorting . Cells are lysed with Nonidet P-40 and the nuclear and cytoplasmic fractions separated by centrifugation . Nuclei are embedded in low-gelling-temperature agarose and the proteinase K and restriction enzyme digestions performed whilst the DNA is immobilized in this form . Total RNA is prepared from the cytoplasmic fraction . This method is simple but reliable and is therefore particularly useful for preparing and analyzing the DNA and RNA from multiple samples when material is limited. J Cell Biol, 1988 Jul, 107(1), 163 - 75 Isolation and characterization of monoclonal antibodies directed against plant plasma membrane and cell wall epitopes: identification of a monoclonal antibody that recognizes extensin and analysis of the process of epitope biosynthesis in plant tissues and cell cultures; Meyer DJ et al.; Membranes from tobacco cell suspension cultures were used as antigens for the preparation of monoclonal antibodies . Use of solid phase and indirect immunofluorescence assays led to the identification of hybridomas producing antibodies directed against cell surface epitopes . One of these monoclonal antibodies (11.D2) was found to recognize a molecular species which on two-dimensional analysis (using nonequilibrium pH-gradient electrophoresis and SDS-PAGE) was found to have a high and polydisperse molecular mass and a very basic isoelectric point . This component was conspicuously labeled by {3H}proline in vivo . The monoclonal antibody cross-reacted with authentic tomato extensin, but not with potato lectin nor larch arabinogalactan . Use of the monoclonal antibody as an immunoaffinity reagent allowed the purification of a tobacco glycoprotein which was identical in amino acid composition to extensin . Finally, immunocytological analyses revealed tissue-specific patterns of labeling by the monoclonal antibody that were identical to those observed with a polyclonal antibody raised against purified extensin . We have concluded that monoclonal antibody 11.D2 recognizes an epitope that is carried exclusively by extensin . Analysis of cellular homogenates through differential and isopycnic gradient centrifugation revealed that biosynthesis of the extensin epitope was found on or within the membranes of the endoplasmic reticulum, Golgi region and plasma membrane . This result is consistent with the progressive glycosylation of the newly-synthesized extensin polypeptide during its passage through a typical eukaryotic endomembrane pathway of secretion . The 11.D2 epitope was not found in protoplasts freshly isolated from leaf tissues . However, on incubation of these protoplasts in appropriate culture media, biosynthesis of the epitope was initiated . This process was not impeded by the presence of chemicals that are reported to be inhibitors of cell wall production or of proline hydroxylation. Blood, 1988 Jul, 72(1), 150 - 8 Ultrastructural localization of the Charcot-Leyden crystal protein (lysophospholipase) to a distinct crystalloid-free granule population in mature human eosinophils; Dvorak AM et al.; The Charcot-Leyden crystal (CLC) protein is a unique constituent of eosinophils and basophils . This protein forms the hexagonal bipyramidal crystals observed in tissues at sites of eosinophil accumulations, possesses lysophospholipase activity (lysolecithin acylhydrolase E.C.3.1.1.5), and comprises an estimated 7% to 10% of total eosinophil protein . The ultrastructural localization of CLC protein was studied in mature peripheral blood eosinophils from normal donors and from patients with the idiopathic hypereosinophilic syndrome . Subcellular localization was evaluated by immunoelectron microscopy using eosinophils, both from buffy coat and purified cell suspensions, that were fixed by a variety of methods . Immunochemical detection of CLC protein employed rabbit antiserum to eosinophil CLC protein, affinity chromatography-purified monospecific IgG antibodies, and postembedding immunogold techniques . Controls for specificity included (1) omission of the primary antibody to CLC protein and (2) substitution of primary antibody with a nonimmune preimmunization serum, a protein A-purified nonimmune IgG, or a protein A-purified nonreactive IgG prepared from solid-phase CLC protein-Sepharose-absorbed anti-CLC antiserum . CLC protein was localized to a minor (approximately 5%) subpopulation of eosinophil granules . These membrane-bound cytoplasmic granules were large (greater than 0.5 mu), were devoid of crystalloid inclusions, and were morphologically compatible with persisting eosinophil primary granules . The crystalloid-containing, large, specific granules did not stain for CLC protein . Insufficient numbers of small dense granules, lipid bodies, and vesiculotubular structures were present to adequately evaluate their potential as additional sites for the subcellular localization of CLC protein . The cellular specificity of the immunogold localization of CLC protein in the eosinophil was affirmed by the absence of staining in neutrophils and lymphocytes present in the same sections . The ultrastructural immunogold localization of CLC protein (lysophospholipase) to a large, crystalloid-free granule in mature circulating eosinophils supports the persistence of a distinct "primary" granule population that serves as a major intracytoplasmic repository for this enzyme. J Biol Chem, 1988 Jun 25, 263(18), 8803 - 9 Purine accumulation in human fat cell suspensions . Evidence that human adipocytes release inosine and hypoxanthine rather than adenosine; Kather H; Human adipocytes are of limited viability (7 +/- 2% release of lactate dehydrogenase/h) and contain active ectophosphatases which are capable of sequentially degrading ATP to adenosine . At densities of 30,000-40,000 cells/ml, human fat cell suspensions accumulated adenosine, inosine, and hypoxanthine, and their concentrations were 38 +/- 8, 120 +/- 10, and 31 +/- 7 nmol/liter after 3 h of incubation . Dipyridamole (10 mumol/liter), an inhibitor of nucleoside transport, caused a 5-7-fold increase in adenosine accumulation which was reduced by 85% on inhibition of ectophosphatases by beta-glycerophosphate and antibodies against ecto-5'-nucleotidase or alpha, beta-methylene 5'-adenosine diphosphate (10 mumol/liter), respectively, indicating that most of the adenosine is produced in the extracellular compartment . Accordingly, the spontaneous accumulation of adenosine was reduced beyond 5 nmol/liter on inhibition of ectophosphatase activities or removal of extracellular AMP by AMP deaminase (4 units/ml) . Added adenosine (30 nmol/liter) disappeared until its concentration approached 5 nmol/liter . Isoproterenol (1 mumol/liter) had no effect on adenosine accumulation regardless whether purine production from extracellular sources was minimized or not . In contrast to adenosine, the concentrations of inosine and hypoxanthine displayed only a modest decrease (30-50%) on inhibition of ectophosphatase activities . In addition, isoproterenol caused a 2-3-fold increase in inosine and hypoxanthine production which was concentration-dependent and could be inhibited by propranolol . It is concluded that the adenosine that accumulates in human adipocyte suspensions is almost exclusively derived from adenine nucleotides which are released by leaking cells . By contrast, inosine and hypoxanthine are produced inside the cells, and the release of these latter purines appears to be linked to ATP turnover via adenylate cyclase. Neurosci Lett, 1988 Jun 17, 89(1), 108 - 13 Fetal serotoninergic transplants in the fourth ventricle of adult rodents reverse sleep disturbances induced by neonatal administration of 5,7-dihydroxytryptamine; McRae-Degueurce A et al.; Rats received a neonatal (at 4 days of age) intracisternal injection of 5,7-dihydroxytryptamine which eliminates serotonin (5-HT) throughout adulthood . Sleep recordings, performed on these rats at adult age, demonstrated significant decreases in paradoxical sleep . Dissociated fetal 5-HT cell suspensions transplanted in the IVth ventricle of these rats restored paradoxical sleep . However, fetal neuronal noradrenergic or cholinergic transplants did not restore paradoxical sleep . These results suggest that paradoxical sleep is directly or indirectly mediated through serotoninergic mechanisms. In Vitro Cell Dev Biol, 1988 Jun, 24(6), 530 - 6 Clonal growth characteristics of adult human prostatic epithelial cells; Peehl DM et al.; The ability of human epithelial cells derived from adult prostatic tissues to undergo clonal growth in culture was examined . In a previously described serum-free culture system, such cells exhibited a density-dependent growth requirement . It was found that raising the level of one of the constituents of the culture medium, bovine pituitary extract, to 100 micrograms/ml permitted excellent clonal growth when as few as 100 cells were inoculated/60-mm2 dish . Raising the levels of supplements other than pituitary extract (cholera toxin, epidermal growth factor, hydrocortisone, or insulin) did not produce this result . The average colony-forming efficiency of cells derived from primary or early passage cultures was approximately 25% . When single cell suspensions were prepared from tissue isolates and directly analyzed for clonal growth, colony-forming efficiencies were approximately 5%, perhaps indicating the proportion of stem cells with proliferative potential in the original isolates . The colony-forming efficiency of a cell population derived from cancer tissue was not significantly different from those of populations derived from normal tissues. Clin Biochem, 1988 Jun, 21(3), 163 - 5 A simpler procedure to study sodium-22 uptake (ouabain insensitive) in human erythrocytes; Gambhir KK et al.; We report a modification of the technique of Mahoney et al . (Blood 1982; 59: 439) for the determination of sodium-22 (22Na+) uptake in human erythrocytes . This modification facilitates the separation of 22Na+ taken up by erythrocytes from the free 22Na+ in the buffer by the addition of dibutyl phthalate, which forms an immiscible layer between the two . To further improve the sensitivity of 22Na+ uptake, we incubated a range of known numbers of erythrocytes with 22Na+ as opposed to the single cell suspension of known hematocrit used in Mahoney's et al . procedure (1) . Erythrocytes are incubated in KCI buffer containing 2627 Bq (0.071 microCi) 22Na+ in a total volume of 0.5 mL for 0.5 h at 37 degrees C . Incubation is terminated by placing the tubes in ice for 10 min and the amount of 22Na+ taken up by the erythrocytes determined . We observe a linear relationship between erythrocyte concentrations (0.5 to 2.5 X 10(9) cells/mL) and percent uptake of 22Na+ (0.37 +/- 0.06 (1 SD) to 1.85 +/- 0.27 (1 SD) of the total 22Na+, respectively) . The procedure is simple and sensitive, and can be used in clinical laboratories for the routine evaluation of 22Na+ uptake in erythrocytes. Biull Eksp Biol Med, 1988 Jun, 105(6), 720 - 3 {Formation of bone tissue by mouse bone marrow cell suspensions in organ culture}; Luriia EA et al.; Adult mouse bone marrow cell suspensions prepared by trypsinization were cultivated in gelatin sponges on millipore filters . When HAWP filters were used, multilayer bone structure was formed . It contained mineralized ground substance, incorporated bone cells and osteoblast layer . With the use of AUFS filters, bone tissue developed not only on the top surface, but also inside the filter. Lab Invest, 1988 Jun, 58(6), 706 - 17 The differentiation potential of tracheal basal cells; Inayama Y et al.; The objective of this study was to examine the differentiation potential of basal cells purified from rabbit tracheas by means of centrifugal elutriation . Cell suspensions containing between 83 and 94% basal cells were inoculated into tracheal grafts denuded of their own epithelium and transplanted to the backs of nude mice . Within 1 week after inoculation, the basal cells reestablished a continuous poorly differentiated epithelial sheet which at 2 to 4 weeks showed typical mucociliary differentiation . Basal cells, ciliated cells, goblet cells, and secretory cells with small granules were present . Some of the latter contained abundant rough endoplasmic reticulum and others contained predominantly smooth endoplasmic reticulum . These data indicate that basal cells can give rise to all major tracheal cell types suggesting that they are the main stem cell in the large airways of rabbits. Radiology, 1988 Jun, 167(3), 825 - 9 Meningeal carcinomatosis in the VX2 rabbit tumor model: detection with Gd-DTPA-enhanced MR imaging; Frank JA et al.; Meningeal carcinomatosis developed in 14 of 14 New Zealand White rabbits after infusion of a VX2 tumor cell suspension into the cisterna magna . All died or were killed 7-15 days after inoculation . Within days of the tumor infusion, magnetic resonance (MR) imaging with gadolinium-diethylenetriaminepentaacetic acid (DTPA) at 0.5 or 1.5 T demonstrated enhancement of the cerebrospinal fluid (CSF) secondary to disruption of the blood-CSF barrier by plaquelike lesions along the meninges . Eventually, meningeal enhancement was observed along the base of the brain and cervical spine . Quantitative assessment of the contrast enhancement on T1-weighted images revealed an increase in mean signal intensity of 213% +/- 130% . Contrast enhancement was not observed in four control animals who received an infusion of cell culture medium . These results demonstrate in an animal model that contrast material-enhanced MR imaging can be used to detect meningeal carcinomatosis by revealing breakdown of the blood-CSF barrier. J Bacteriol, 1988 Jun, 170(6), 2676 - 82 Changes induced in the permeability barrier of the yeast plasma membrane by cupric ion; Ohsumi Y et al.; A specific effect of Cu2+ eliciting selective changes in the permeability of intact Saccharomyces cerevisiae cells is described . When 100 microM CuCl2 was added to a cell suspension in a buffer of low ionic strength, the permeability barrier of the plasma membranes of the cells was lost within 2 min at 25 degrees C . The release of amino acids was partial, and the composition of the amino acids released was different from that of those retained in the cells . Mostly glutamate was released, but arginine was mainly retained in the cells . Cellular K+ was released rapidly after CuCl2 addition, but 30% of the total K+ was retained in the cells . These and other observations suggested that Cu2+ caused selective lesions of the permeability barrier of the plasma membrane but did not affect the permeability of the vacuolar membrane . These selective changes were not induced by the other divalent cations tested . A novel and simple method for differential extraction of vacuolar and cytosolic amino acid pools by Cu2+ treatment was established . When Ca2+ was added to Cu2+-treated cells, a large amount of Ca2+ was sequestered into vacuoles, with formation of an inclusion of a Ca2+-polyphosphate complex in the vacuoles . Cu2+-treated cells also showed enhanced uptake of basic amino acids and S-adenosylmethionine . The transport of these substrates showed saturable kinetics with low affinities, reflecting the vacuolar transport process in situ . With Cu2+ treatment, selective leakage of K+ from the cytosolic compartment appears to create a large concentration gradient of K+ across the vacuolar membrane and generates an inside-negative membrane potential, which may provide a driving force of uptake of positively charged substances into vacuoles . Cu2+ treatment provides a useful in situ method for investigating the mechanisms of differential solute pool formation and specific transport phenomena across the vacuolar membrane. J Neuroimmunol, 1988 Jun, 18(3), 217 - 22 Failure to detect the presence of pluripotential haemopoietic stem cells in the mouse brain; Stedra J et al.; Single cell suspensions prepared from adult mouse brains were tested for the presence of pluripotential haemopoietic stem cells (colony-forming units, CFU) by transfer into an irradiated recipient and enumeration of the CFU in the recipient's spleen . In contrast to the findings of others (Bartlett, 1982), we did not detect CFU after injection of brain cell suspensions, although they were detectable after inoculation with bone marrow cells . The number of CFU in recipients after transfer of increasing numbers of brain cells was the same as that detected in the irradiated controls which had not received any transferred cells . Finally, cells from the brain, in contrast to bone marrow cells, were not able to protect recipient animals from the effects of lethal irradiation. Cancer Res, 1988 Jun 1, 48(11), 3040 - 4 Tissue uptake, distribution, and potency of the photoactivatable dye chloroaluminum sulfonated phthalocyanine in mice bearing transplantable tumors; Chan WS et al.; The potency of chloroaluminum sulfonated phthalocyanine (ClAlSPc) as a photosensitizing agent for photodynamic therapy of cancer was evaluated in vivo by its ability to be taken up and retained by murine tumors of diverse histological origin . Antitumor effects following laser irradiation were evaluated by measurement of the tumor weights of dissected-out tumor masses . Three tumors (Colo 26, a colorectal carcinoma; M5076, a reticulum cell sarcoma; and UV-2237, a fibrosarcoma) growing s.c . in the flank region retained substantially greater quantities of ClAlSPc than did adjacent skin and muscle achieving peak values 24-48 h after the i.v . administration of ClAlSPc (10 mg/kg) . The relative magnitude of ClAlSPc retention by these tumors was Colo 26 greater than M5076 greater than UV-2237 . However, normal liver and spleen were organs which retained the greatest amounts of ClAlSPc even compared to the s.c . grown tumors and other normal tissues examined . Flow cytometric analysis of tumor cell suspensions obtained from collagenase-digested tumors showed that individual neoplastic cells were capable of taking up and retaining ClAlSPc . Photodynamic therapy, undertaken by i.v . administration of dye (5 mg/kg) followed 24 h later by local laser light irradiation (675 nm, 100 J), brought about significant (Colo 26, M5076, and 3LL tumors) and obvious but nonsignificant (UV-2237 tumor) reductions in tumor weights, as assessed 5 days later . Thus, selective tumor retention of ClAlSPc coupled with a significant response to red light produced dramatic alterations in cancer growth. Int J Radiat Biol Relat Stud Phys Chem Med, 1988 Jun, 53(6), 921 - 33 Variations in the spectrum of lesions produced in the DNA of cells from mouse tissues after exposure to gamma-rays in air-breathing or in artificially anoxic animals; Murray D et al.; Gamma-ray-induced DNA-protein crosslinks (dpc) are preferentially induced in cultured cells irradiated at very low oxygen tensions (Meyn et al . 1987) . Since some cells within mouse tumors may be radiobiologically hypoxic, dpc may also be induced in such cells after irradiation in vivo . To examine this possibility, mice bearing either an FSa or NFSa fibrosarcoma in their hind legs were whole-body irradiated either while breathing atmospheric oxygen or 15 min after cervical dislocation, which induces uniform anoxia . DNA single-strand breaks (ssb) and dpc were then assayed both in tumors and normal tissues by alkaline elution . The level of dpc was inferred from the observed increase in ssb yield after digestion of the cell lysates with proteinase K . In addition, cell suspensions were irradiated in vitro, on ice, exposed to atmospheric oxygen tensions . Few dpc were detected in the DNA from tumor cells irradiated in vitro; however, in cells from both FSa and NFSa tumors irradiated in situ there was a significant level of protein-concealed ssb, and thus of dpc . These data are most likely the result of the relative hypoxia of a proportion of cells from both the FSa and NFSa tumor in the air-breathing animals . Induction of dpc was further enhanced in the DNA from tumor cells irradiated under anoxic conditions . A significant level of dpc was also observed in jejunal and spleen cells irradiated in vivo; however, since a significant level of protein-concealed breaks was also observed in cells irradiated in vitro, oxygenation appears not to be the only parameter capable of modifying the proportion of protein-concealed ssb, and the effects of proteinase K on the DNA elution rate for normal mouse tissues may be complex. J Histochem Cytochem, 1988 Jun, 36(6), 679 - 83 Sensitive detection of immunogold-silver staining with darkfield and epi-polarization microscopy; De Waele M et al.; We evaluated the contribution of darkfield and epi-polarization microscopy to the detection of leukocyte cell surface antigens with immunogold-silver staining (IGSS) . Lymphocyte cell surface differentiation antigens were labeled with monoclonal antibodies and IGSS as described for brightfield microscopy . In darkfield and epi-polarization microscopy the labeling appeared as bright spots on a dark background . The sensitivity of detection was much higher than that of brightfield microscopy . Sixteenfold higher dilutions of the monoclonal antibody could be used to detect all cells expressing the antigen in the cell suspension . However, non-specific staining was also better visualized . The latter could be reduced to a level comparable to that of brightfield microscopy only by use of weaker labeling conditions . A 25% reduction of the silver enhancement time was necessary for this purpose . However, these weaker labeling conditions also reduced the intensity of the specific staining . Therefore, the efficiency of IGSS, as detected with darkfield and epi-polarization microscopy, was only fourfold greater than that found with brightfield microscopy or that of an immunofluorescence procedure . Especially in combination with transmitted light, to improve cell identification, epi-polarization microscopy is a reliable and sensitive method for detection of immunogold-silver-labeled cell surface antigens for diagnostic and research purposes. Cell Biol Toxicol, 1988 Jun, 4(2), 211 - 23 A novel class of unstable 6-thioguanine-resistant cells from dog and human kidneys; Turker MS et al.; Thioguanine-resistant primary clones were grown from single cell suspensions obtained from dog and human kidneys by enzymatic digestion . In medium containing a relatively high concentration (10 micrograms/ml) of thioguanine, thioguanine-resistant primary clones arose from each source at frequencies ranging from 10(-4) to 10(-5) . A reduction in total hypoxanthine uptake was found in the thioguanine-resistant primary clones which had developed in thioguanine medium, consistent with a reduction in hypoxanthine phosphoribosyltransferase activity . When these thioguanine-resistant primary clones were subsequently grown in the absence of thioguanine and assayed for the thioguanine-resistant phenotype and hypoxanthine phosphoribosyltransferase activity, it was found that most were now thioguanine-sensitive and yielded cell-free extracts with substantial amounts of hypoxanthine phosphoribosyltransferase activity . In contrast, thioguanine-resistant human clones grown continuously in the presence of thioguanine yielded cell-free extracts with little or no detectable hypoxanthine phosphoribosyltransferase activity . Southern blot analysis demonstrated no structural alterations in the hypoxanthine phosphoribosyltransferase gene in thioguanine-resistant primary human kidney clones . These results suggest that a novel mechanism(s) for thioguanine resistance and the control of hypoxanthine phosphoribosyltransferase expression may occur in dog and human kidney cells. J Neurocytol, 1988 Jun, 17(3), 351 - 60 Transplantation of oligodendrocytes and Schwann cells into the spinal cord of the myelin-deficient rat; Duncan ID et al.; Transplantation of oligodendrocytes or Schwann cells into the spinal cord of the newborn myelin-deficient (md) rat, an X-linked myelin mutant, was carried out and the extent of myelination of CNS axons studied . Dissociated glial cell suspensions, prepared from the spinal cords of female litter-mates, were injected into the lumbar spinal cord of 15 md rats and 5 normal litter-mates . In eight of the md rats examined 12 to 21 days post-transplantation patches of myelin produced by the transplanted oligodendrocytes were found in the dorsal or ventral columns . In two rats, small patches of myelination were found in more than one site . The myelin in these patches was positive on immunocytochemical staining for proteolipid protein . These observations were interpreted as evidence of the origin of this myelin from donor oligodendrocytes, as the md rat has an abnormality in synthesis of this protein . In addition, this myelin differed in its ultrastructure from host myelin, having a normal intraperiod line . Injection of cultured Schwann cells also resulted in extensive myelination of axons in the dorsal columns by these cells. Appl Environ Microbiol, 1988 Jun, 54(6), 1330 - 3 Continuous-sterilization system that uses photosemiconductor powders; Matsunaga T et al.; We report a novel photochemical sterilization system in which Escherichia coli cells were sterilized with photosemiconductor powders (titanium oxide) . For sterilization that could be used in practice, it was necessary to separate the TiO2 powders from the cell suspension . Therefore, semiconductor powders were immobilized on acetylcellulose membranes . We constructed a continuous-sterilization system consisting of a TiO2-immobilized acetylcellulose membrane reactor, a mercury lamp, and a masterflex pump . As a result, under the various sterilization conditions examined, E . coli (10(2) cells per ml) was sterilized to less than 1% survival when the cell suspension flowed in this system at a mean residence time of 16.0 min under irradiation (1,800 microeinsteins/m2 per s) . We found that this system was reusable. Immunol Invest, 1988 Jun, 17(4), 309 - 19 Flow cytometric analysis of lymphocyte activation in the mixed lymphocyte response; Prince HE et al.; Interleukin 2 receptor (IL2R) expression may be a useful parameter for assessing lymphocyte activation in the mixed lymphocyte response (MLR) . However, the contribution of irradiated stimulator (S*) cells to the levels of IL2R+ cells recovered must first be defined . We have used a flow cytometric parameter termed the sip count to assess this potential contribution of S* cells . This parameter, which is the number of cells within a defined cell gate, is a reflection of the viable cell number per culture well, since (a) a constant number of cells were plated per well on day 0, (b) cells recovered from a well were resuspended in a constant volume, (c) the flow cytometer aspirated (sipped) a constant volume of cell suspension, and (d) nonviable cells were not included in the gate . Sip count assessment showed that only 5% of S* cells were recoverable by day 4 of culture; in contrast, 70% or more of unstimulated responder cells were recoverable . Sip counts of MLR cultures identified an increase in cell number beginning on day 5, reflecting DNA synthesis and cell division . We then used the sip count to assess changes in the levels of IL2R+ cells in MLR cultures . The number of IL2R+ cells continued to increase up to day 7, even though maximal DNA synthesis occurred on day 5 . Further, dual color analysis revealed that the proportion of CD4 cells expressing IL2R was maximal on day 5, whereas the proportion of CD8 cells expressing IL2R continued to increase until day 9 . These findings show that flow cytometry can be used to study lymphocyte activation by alloantigens. J Neurol Sci, 1988 Jun, 85(2), 197 - 207 Myogenicity in vitro and in vivo of mouse muscle cells separated on discontinuous Percoll gradients; Morgan JE; Mouse muscle cells, obtained by enzymatically disaggregating newborn mouse muscle, were separated on a discontinuous Percoll gradient . The myogenicity in vitro of the resultant cell fractions was examined by counting the percentage of nuclei in myotubes . Myogenicity in vivo was assessed by implanting a cell suspension of one of the allotypes of glucose-6-phosphate isomerase (GPI) into a regenerating skeletal muscle graft of a second GPI allotype: the finding of hybrid GPI indicated that the implanted cells were myogenic . Separation of mouse muscle cells on a discontinuous Percoll gradient gave rise to two myogenic fractions, one of which was more myogenic in vitro than were the unseparated cells and one of which was less myogenic . Both of these fractions were myogenic in vivo . A cell fraction was also produced which was non-myogenic in vitro as well as in vivo . In vitro and in vivo measurements of myogenicity were therefore in broad agreement. Biochem Biophys Res Commun, 1988 May 31, 153(1), 203 - 8 Human B and T lymphocytes convert leukotriene A4 into leukotriene B4; Odlander B et al.; Incubation of human tonsillar B lymphocytes and peripheral blood T lymphocytes with leukotriene A4 led to the formation of leukotriene B4 . The purity of these cell suspensions was more than 99%, containing less than 0.5% monocytes . Incubation of purified B or T lymphocytes with the calcium ionophore A23187 did not lead to the formation of any detectable amounts of leukotrienes . Several established cell lines of B and T lymphocytic origin were also found to convert leukotriene A4 into leukotriene B4, showing that monoclonal lymphocytic cells possess leukotriene A4 hydrolase activity. Eur J Biochem, 1988 May 16, 174(1), 189 - 97 Methanogenesis and ATP synthesis in methanogenic bacteria at low electrochemical proton potentials . An explanation for the apparent uncoupler insensitivity of ATP synthesis; Kaesler B et al.; The rate of methane formation from H2 and CO2, the intracellular ATP content and the electrochemical proton potential (delta mu H+) were determined in cell suspensions of Methanobacterium thermoautotrophicum, which were permeabilized for K+ with valinomycin (1.2 mumol/mg protein) . In the absence of extracellular K+ the cells formed methane at a rate of 4 mumol min-1 (mg protein)-1, the intracellular ATP content was 20 nmol/mg protein and the delta mu H+ was 200 mV (inside negative) . When K+ was added to the suspensions the measured delta mu H+ decreased to the value calculated from the {K+}in/{K+}out ratio . Using this method of delta mu H+ adjustment, it was found that lowering delta mu H+ from 200 mV ({K+}in/{K+}out = 1000) to 100 mV ({K+}in/{K+}out = 40) had no effect on the rate of methane formation and on the intracellular ATP content . At delta mu H+ values below 100 mV ({K+}in/{K+}out less than 40) both the rate of methanogenesis and the ATP content decreased . Methanogenesis completely ceased and the ATP content was 2 nmol/mg when delta mu H+ was adjusted to values lower 50 mV ({K+}in/{K+}out less than 7) . The data show that methanogenesis from H2 and CO2 and ATP synthesis in M . thermoautotrophicum are possible at relatively low electrochemical proton potentials . Similar results were obtained with Methanosarcina barkeri . Protonophoric uncouplers like 3,5,3',4'-tetrachlorosalicylanilide (TCS) or 3,5-di-tert-butyl-4-hydroxy-benzylidenemalononitrile (SF 6847) were found not to dissipate delta mu H+ below 100 mV in M . thermoautotrophicum even when used at high concentrations (400 nmol/mg protein) . This finding explains the observed uncoupler insensitivity of methanogenesis and ATP synthesis in this organism. Arch Biochem Biophys, 1988 May 15, 263(1), 191 - 8 Induction of phytoalexin synthesis in soybean: enzymatic cyclization of prenylated pterocarpans to glyceollin isomers; Welle R et al.; A microsome preparation from elicitor-challenged soybean cell suspension cultures catalyzed an NADPH-dependent and oxygen-dependent cyclization of a mixture of 2- and 4-dimethylallylglycinols to the glyceollin isomers I-III . This is the last committed step in glyceollin biosynthesis . The cyclase was inhibited in a light-reversible manner by carbon monoxide in the presence of oxygen . Cyclase was also inhibited by cytochrome c, NADP+, and a number of inhibitors of cytochrome P-450 enzymes . NADH in the presence of low concentrations of NADPH had a synergistic effect . On a Percoll gradient, the position of cyclase coincided with marker enzymes for the endoplasmic reticulum . These properties identify the cyclase as a cytochrome P-450-dependent monooxygenase . Unstimulated soybean cell culture did not contain detectable cyclase activity . Challenge with either a glucan elicitor from Phytophthora megasperma f.sp . glycinea or with yeast extract caused strong stimulation of cyclase activity with a maximum at about 24 h after elicitor addition. Cancer Res, 1988 May 15, 48(10), 2779 - 83 Effects of hyperoxia on growth characteristics of metastatic murine tumors in the lung; Margaretten NC et al.; Suspensions of an oxygen-sensitive (MT-7) and of an oxygen-insensitive(M109) tumor cell line were injected i.v . into BALB/c mice . Exposure to 100% O2 after injection of the cells did not modify the initial arrest of either cell line in the lung . Exposure of animals given injections of MT-7 cells for 60 h to 100% oxygen decreased the number of lung colonies formed even when onset of oxygen exposure was delayed up to 10 days after injection of the cell suspension . Cell cycle time and growth fraction in lung colonies growing in vivo were estimated from an analysis of the percentage of mitoses labeled . In lung colonies formed by MT-7 cells, hyperoxia produced a mitotic delay and a 30 to 40% reduction in the growth fraction . In M109-derived colonies, oxygen did not change cell cycle times or reduce growth fraction . In earlier experiments done in vitro and reported by others it had been found that, in tumor cell lines other than the ones used in the present study, a prolongation of the early prophase was the most oxygen-sensitive event . The present data show that in vivo oxygen inhibits lung colony formation in MT-7 cells by a similar mechanism. J Immunol, 1988 May 15, 140(10), 3541 - 6 Granulocyte-macrophage colony-stimulating factor modulates the excitation-response coupling sequence in human neutrophils; Naccache PH et al.; We have investigated the effects of granulocyte-macrophage (GM) CSF on two biochemical responses thought to play internal signaling roles in the neutrophils, namely the increase in the concentration of free calcium and the changes in the internal pH . The changes in the right-angle light scatter of the cell suspensions were also examined . GM-CSF was found not to affect the resting levels of calcium or the internal pH of the cells . However, pre-incubation of the neutrophils with the growth factor resulted in an increase in the magnitude of the calcium transients that follow the stimulation of the cells with chemotactic factors as well as a profound depression of the cell alkalinization that is induced by fMet-Leu-Phe, leukotriene B4, and platelet-activating factor, as well as by phorbol esters . The rapid cytoplasmic acidification elicited by these agents was apparently magnified . GM-CSF did not directly affect the Na+/H+ antiport as GM-CSF-treated cells were as capable as untreated cells of recovering from an acid load . The right-angle light scatter responses of the cells to the chemotactic factors were found not to be affected by GM-CSF . These results provide an initial description of the effects of GM-CSF on the cellular physiology of the neutrophils and insights into the mechanism of action of this factor as well as into the excitation-response coupling sequence activated by chemotactic factors. J Natl Cancer Inst, 1988 May 4, 80(5), 351 - 60 Mixed phenotype lymphomas of B10.H-2a-H-4bp/Wts mice: characterization of component cell populations; Willoughby PB et al.; Three murine lymphomas of H-2a origin were investigated . Each of these lymphomas was derived in B10.H-2aH-4bp/Wts (2a4b) mice after a hyperimmunization protocol involving sheep red blood cells . Analysis of cell suspensions derived from the splenic tumors revealed a mixed lymphoid phenotype of the total cell population . Both the T-cell and B-cell populations remained after multiple passages of the tumors in syngeneic mice . A multiparameter approach combining immunologic and molecular genetic techniques was taken to determine the malignant cell population within these lymphomas . This analysis included depletion experiments utilizing histocompatible (B10.A X 21M)F1 mice to distinguish normal host-derived cells from the malignant tumor cells, fluorescence-activated cell sorting of the two cell populations followed by in vivo growth of the sorted cells, in vitro and in vivo cloning attempts, and molecular analysis of genomic DNA derived from the splenic tumors for clonal rearrangements of the immunoglobulin heavy-chain and T-cell receptor loci . Each of the three lymphomas was shown to be a malignant B-cell tumor that has an associated nonneoplastic T-cell component . The nature and functional significance of the numerous T-cells present in the B-cell lymphomas remain unknown and are being investigated. J Surg Res, 1988 May, 44(5), 473 - 8 A possible mechanism for the release of serotonin from the gut caused by pentagastrin; Gronstad KO et al.; Pentagastrin (PG) is a potent agent causing release of serotonin (5-HT) from patients with carcinoid tumors . The physiological release of 5-HT from gut enterochromaffin cells is controlled by beta-adrenoceptors . Studies on carcinoid tumor cell suspensions, acute or in culture, have shown that catecholamines (CA), but not PG, release 5-HT, thus indicating an indirect mode of action by the peptide . In this study the mechanism for release of 5-HT from the gut induced by PG was investigated in animal models . The test protocol for patients was used in anesthetized cats . Portal blood samples were drawn after PG injection (0.6 microgram/kg iv), which resulted in significantly increased levels of 5-HT at 3 and 5 min postinjection . The PG-induced release was totally inhibited after blockade of beta-adrenoceptors (propranolol) or of slow calcium channels (verapamil) as well as after adrenalectomy . Blockade of beta-adrenoceptors or slow calcium channels decreased the basal levels of 5-HT, while adrenalectomy caused no change . In separate experiments CA were measured after PG injection in caval blood, drawn at the level of the adrenal veins . There was a significant increase in the levels of dopamine and epinephrine postinjection, while the levels of norepinephrine were stable . The changes of CA levels after PG injection could be prevented by adrenalectomy . These results further suggest an indirect action of PG in the release of 5-HT from the feline gut via activation of beta-adrenoceptors by epinephrine released from the adrenals. J Cell Physiol, 1988 May, 135(2), 262 - 8 Growth characteristics of human epidermal melanocytes in pure culture with special reference to genetic differences; Hirobe T et al.; Human melanocyte cultures were established using disaggregated epidermal cell suspensions derived from foreskins and plated onto culture dishes in medium containing 2% fetal bovine serum, growth factors, hormones, and melanocyte growth factor (MGF) extracted from bovine hypothalamus (Wilkins et al., J.Cell . Physiol., 122:350, 1985) . After 2 days in culture the cells were transferred to serum-free medium to eliminate keratinocyte and fibroblast growth . Melanocytes grew preferentially and pure melanocyte populations could be harvested after 12-16 days in vitro . Melanocytes were later subcultured in the presence of 1% FBS . Pure melanocyte cultures were characterized by light and electron microscopic criteria, as well as by cytochemical demonstration of the melanocyte-specific enzyme, tyrosinase . At the ultrastructural level, cultured melanocytes derived from black (negroid) neonatal skin (B-M) had numerous mature rod-shaped stage IV melanosomes, while white (caucasoid) skin-derived melanocytes (W-M) in culture contained no mature melanosomes . Growth rate, cell yield, and in vitro lifespan for B-M were more than twice that for W-M in pure melanocyte cultures in the presence of MGF . Our results suggest that MGF-dependent growth of B-M differs from that of W-M. Blut, 1988 May, 56(5), 221 - 7 Sulfide silver amplification of ferritin iron cores in blood and bone marrow cells . Methods, adaptations to microphysical analyses, and the impact of advanced iron overload; Hausmann K et al.; A short exposure of cell suspensions to gaseous hydrogen sulfide, appropriate fixations, and subsequent physical development of silver shells around sulfidated insoluble metals were used to amplify ferritin iron cores in blood and bone marrow cells . The methods described are suitable for both light microscopy and transmission electron microscopy . These techniques made it possible to visualize Prussian Blue stainable ferritin and haemosiderin, as well as a large variety of isoferritin iron and other smaller particles beyond the sensitivity of Prussian Blue staining . Admixtures of sulfidatible zinc and traces of other heavy metals had to be taken into consideration . For further research, adaptations of sulfide silver staining to microphysical analyses were developed . However, conventional energy dispersive X-ray analysis was not sensitive enough to signalize the presence of Fe in sulfide silver amplified iron cores of a single or a few ferritin molecule(s) . Proton-induced X-ray emission was used to measure Fe and Zn down to 1 fg/single cell in unstained or sulfide silver stained smears on thin foils . However, multielement analysis of homogeneous cell concentrates was much easier to perform and far more sensitive . In advanced iron overload, highly increased sulfide silver staining was found in peripheral blood cells including lymphocytes, monocytes, eosinophils, basophils, and--in extreme cases--also in neutrophils and platelets. Ultrasonics, 1988 May, 26(3), 168 - 70 Ultrasonic attenuation in red blood cell suspensions; Dai HP et al.; A theoretical analysis of ultrasonic attenuation in aqueous suspensions of red blood cells was carried out . This verified the relative importance of several mechanisms: absorption accounts for 60% of the attenuation, viscous relative motion loss accounts for less than 30%, and the sound scattering contribution is negligible. Arch Biochem Biophys, 1988 May 1, 262(2), 445 - 54 Purification and properties of S-adenosyl-L-methionine:nicotinic acid-N-methyltransferase from cell suspension cultures of Glycine max L; Upmeier B et al.; A soluble enzyme which catalyzes the transfer of the methyl group from S-adenosyl-L-methionine to the nitrogen atom of pyridine-3-carboxylic acid (nicotinic acid) could be detected in protein preparations from heterotrophic cell suspension cultures of soybean (Glycine max L.) . Enzyme activity was enriched nearly 100-fold by ammonium sulfate precipitation, gel filtration, and ion-exchange chromatography to study kinetic properties . S-adenosyl-L-methionine:nicotinic acid-N-methyltransferase (EC 2.1.1.7) showed a pH optimum at pH 8.0 and a temperature optimum between 35 and 40 degrees C . The apparent KM values were determined to be 78 microM for nicotinic acid and 55 microM for the cosubstrate . S-Adenosyl-L-homocysteine was a competitive inhibitor of the methyltransferase with a KI value of 95 microM . The native enzyme had a molecular mass of about 90 kDa . The catalytic activity was inhibited by reagents blocking SH groups, whereas other divalent cations did not significantly influence of the enzyme reaction . The purified methyltransferase revealed a remarkable specificity for nicotinic acid . No other pyridine derivative was a suitable methyl group acceptor . To study a potential methyltransferase activity with nicotinamide as substrate, an additional purification step was necessary to remove nicotinamide amidohydrolase activity from the enzyme preparation . This was achieved by affinity chromatography on S-adenosyl-L-homocysteine-Sepharose thus leading to a 580-fold purified enzyme which showed no methyltransferase activity toward nicotinamide as substrate. Immunology, 1988 May, 64(1), 147 - 53 A role for mature B cells in bone marrow transplantation; Yoshio T et al.; The proliferative potential of membrane Ig (mIg)-bearing B lymphocytes was assessed in an adoptive transfer system based on the use of non-inbred rabbits matched for major histocompatibility (MHC) antigens and mismatched for immunoglobulin (Ig) allotypes . Cell suspensions made from spleens (SP), mesenteric lymph nodes (LN), or bone marrow (BM) of allotype b4b5 rabbits were deprived of B cells with mIg of the b4 type by adherence to plastic dishes coated with affinity-purified anti-b4 . When such b4-depleted cell populations were injected into newborn hosts of allotype b6b6, stable and lasting chimerism promptly resulted, in which donor-derived products were almost entirely of the b5 allotype . Chimeras formed by transfer of unfractionated cells from b4b5 donors, on the other hand, exhibited a predominance of the b4 allotype, as seen in the living donors . BM but not SP or LN contained precursors capable of differentiating into mIg+ lymphocytes in culture, but no evidence was obtained for engraftment and differentiation by such B-cell precursors or more primitive stem cells in vivo . These studies suggest a potentially significant role for mature B cells in reconstituting the immune system of human transplant recipients. J Chem Neuroanat, 1988 May-Jun, 1(3), 133 - 46 Laminin facilitates and guides fiber growth of transplanted neurons in adult brain; Zhou FC et al.; Laminin has been shown in vitro to act as a surface adhesive molecule for neuronal process elongation . To test whether laminin has a similar role in the brain, we sequentially injected laminin and transplanted fetal neurons into various brain regions to determine if the fetal neurons would preferentially grow along a laminin injection tract . In the fetal brain, the raphe area of the rostral rhombencephalon is rich in serotonergic (5-HT) neurons; the rostral ventral mesencephalon is rich in dopamine (DA) neurons, while the lateral rhombencephalon is rich in norepinephrinergic (NE) neurons . These three areas were transplanted to the motor cortex, neostriatum or hippocampus of adult animals . The tract used for microinjection of cell suspension was then immediately filled with laminin in a suspension media or a laminin-collagen (type IV) mixture . In other animals, laminin or control solution was injected in a separate needle tract displaced 0.3-1 mm from the transplant injection tract . Straight and thick 5-HT, DA or NE immunoreactive (IR) fibers (stained with anti-5-HT or anti-tyrosine hydroxylase antiserum) were predominant within the laminin-treated tracts, or were directed toward the laminin-treated parallel tracts when it was positioned less than 0.5 mm from the transplant site . The density of 5-HT-, DA- and NE-IR fibers in the injection tracts in all three brain areas was much higher for laminin and laminin-collagen mixture than control media . Thin axonal fibers of fetal 5-HT and NE neurons were observed surrounding the laminin-treated tracts, but not around vehicle-injected tracts . In addition, a number of transplanted 5-HT, DA and NE neuronal cell bodies were seen within the laminin-treated tracts, but not in vehicle-treated tracts . Finally, laminin injection to the hippocampus, motor cortex or neostriatum of the adult brain did not stimulate sprouting of undamaged adult 5-HT or NE fibers . These results suggest that purified laminin can facilitate and guide process outgrowth of 5-HT, DA and NE neurons during early developmental stage, but does not induce sprouting on these same fiber types in the adult brain. Biull Eksp Biol Med, 1988 May, 105(5), 621 - 3 {Structural and immunologic indicators of Peyer's patches in the human fetus}; Khlystova ZS et al.; Histological and immunological methods were used