Mol Microbiol, 1987 Sep, 1(2), 151 - 8 Heat shock 70 gene is differentially expressed in Histoplasma capsulatum strains with different levels of thermotolerance and pathogenicity; Caruso M et al.; The response to heat shock has been examined in two strains of the dimorphic pathogenic fungus Histoplasma capsulatum, which differ considerably in thermotolerance and pathogenicity . The gene for the 70 kD heat shock protein (hsp70) was isolated using a Drosophila hsp70 gene to screen a cosmid library of the DNA from the temperature-sensitive Downs strain (low level of thermotolerance for mice) . Using the cloned gene as a probe, we have measured the transcription of the endogenous hsp70 gene at 25 degrees C and in response to temperature shift to 34 degrees, 37 degrees and 40 degrees C, temperatures that trigger the mycelial to yeast phase transition in this fungus . The gene is constitutively transcribed at low levels, both in the yeast and the mycelial stages . Synthesis of hsp70 mRNA was transiently increased 1 to 3 h after the temperature shifts . By Northern analysis, peak levels of transcription were shown to occur at 34 degrees C in the Downs strain and at 37 degrees C in the more pathogenic G222B strain . Our results are consistent with reports in which it has been shown that heat shock gene expression is part of temperature adaptation and probably developmental processes . The low levels of transcription of the hsp70 gene in the Downs strain at 37 degrees C correlate with its greater temperature sensitivity and low level of virulence. Mutagenesis, 1987 Sep, 2(5), 357 - 65 Hypothesis: some mutagens directly alter specific chromosomal proteins (DNA topoisomerase II and peripheral proteins) to produce chromosome stickiness, which causes chromosome aberrations; Gaulden ME; Recent biochemical and molecular biological data on the composition and structure of the chromosome and the nucleus, combined with observations on the chromosomes of mutant yeast cells and grasshopper neuroblasts, offer new perspectives on mutagen-induced chromosome stickiness and its relation to chromosome breakage . A hypothesis consistent with these data states that chromosome stickiness (i) results from changes in specific non-histone proteins (topoisomerase II and the peripheral proteins) that are integral components of the chromosome and whose function is necessary for separation and segregation of chromatids, the changes being caused either by mutation in structural genes for the proteins (heritable stickiness) or by direct action of mutagens on the proteins (induced stickiness); (ii) occurs in various degrees (slight, moderate, severe, extreme) that are determined by the number of target protein molecules affected, a certain number (threshold) of affected molecules at a given site on a chromosome being required to resist the forces of anaphase movement in order to produce microscopically detectable stickiness; (iii) results from molecular events that can occur at several phases of the cell cycle (including interphase), but can only be recognized at prometaphase, metaphase and anaphase; and (iv) causes chromosome aberrations by the physical stretching and breaking of chromatids at the sticky sites; hence the breakage resulting from stickiness is a secondary effect that requires anaphase movement, in contrast to breakage resulting from direct action of mutagens on DNA. Br J Haematol, 1987 Sep, 67(1), 3 - 10 Development of the phagocytic and cidal capacity during maturation of myeloid cells: studies on cells from patients with chronic myelogenous leukaemia; Segel EK et al.; Myeloid cells from peripheral blood of patients with chronic myelogenous leukaemia were isolated and fractionated by density gradient centrifugation using Lymphoprep gradient followed by discontinuous Percoll gradients . Six fractions were obtained, each enriched in one of the morphologically identifiable types of myeloid cells from myeloblasts to polymorphonuclear neutrophils . Each of these cell types were functionally and biochemically characterized . The development of the capacities for phagocytosis and killing of yeast cells and the ability to generate a respiratory burst of phagocytosis correlated closely with the content of cytochrome b and vitamin B12-binding protein, a marker of specific granules . These results support the notion that the specific granules provide the developing neutrophil with components which are essential for its microbicidal activity. J Immunol, 1987 Sep 1, 139(5), 1679 - 82 Antigenic analysis of the repeat domain of the circumsporozoite protein of Plasmodium vivax; Romero P et al.; In the present study we analyzed the fine specificity of mouse monoclonal and human polyclonal antibodies directed against the repeat domain of the circumsporozoite (CS) protein of the human malaria parasite, Plasmodium vivax . Five synthetic peptides, representing monomeric and dimeric repeats of this malarial antigen, were assayed for their capacity to inhibit the binding of these antibodies to a yeast-derived recombinant CS protein . The results revealed the existence of at least two distinct repeated overlapping epitopes in the CS protein of P . vivax . Furthermore, polyclonal sera contain antibodies which recognize additional determinants not represented by the synthetic repeat peptides . Some of these sera contain antibodies recognizing a region flanking the repeat domain (region I) . The present findings are in contrast with the antibody response in rodents and humans to the Plasmodium falciparum CS protein, which is directed against a single repeated immunodominant epitope. Schweiz Med Wochenschr, 1987 Aug 29, 117(35), 1289 - 96 {Disseminated histoplasmosis due to Histoplasma capsulatum in a patient with acquired immunodeficiency syndrome (AIDS)}; Dietrich PY et al.; Very commonly encountered in the United States, histoplasmosis is rare in Europe, where only 27 patients have so far been infected by this mycosis . In Africa, two varieties of histoplasmosis have been observed: those due to H . capsulatum and H . duboisii . Histoplasmosis due to H . capsulatum is one of the twelve secondary infectious diseases listed in the surveillance definitions of AIDS . This complication has been described only in approximately 20 patients with AIDS; all patients had stayed on the American Continents . We report the case of a 30-year-old African male who lived in Switzerland and in Zaire . With AIDS and multiple Kaposi's sarcoma, the patient died from disseminated histoplasmosis due to H . capsulatum; a peripheral blood smear obtained a few hours before death revealed numerous typical yeast forms of H . capsulatum inside polymorphonuclear leukocytes . Post-mortem examination and cultures confirmed the diagnosis of disseminated histoplasmosis . Histoplasmosis should be suspected in AIDS patients even in Europe, especially where they have stayed in endemic areas . Examination of blood smears and bonemarrow aspirate may allow early recognition of the disease and permit appropriate treatment with amphotericin B and ketoconazole. Arch Biochem Biophys, 1987 Aug 15, 257(1), 17 - 26 Inhibition of hexokinase activity by a fructose 2,6-bisphosphate-dependent cytosolic protein from liver; Niemeyer H et al.; Mammalian and yeast hexokinases were found to be reversibly inhibited by fructose 2,6-bisphosphate, an effect requiring the presence of a cytosolic protein factor . Experimental evidence suggests that this factor (inhibitor) is a regulatory protein, the interactions of which with hexokinases are modulated by fructose 2,6-bisphosphate . The Vmax of hexokinase D was decreased, and no changes on other kinetic parameters were observed . The inhibitor was present in fresh liver cytosol filtered through Sephadex G-25 and was partially isolated by negative absorption on DEAE-cellulose followed by ammonium sulfate fractionation . The inhibitor was also present in brain and kidney, but not in muscle . A molecular mass of 200,000 was determined by gel filtration . The inhibition was dependent on the concentrations of both the inhibitory protein and fructose 2,6-bisphosphate . No delay in fructose 2,6-bisphosphate inhibition was observed . Several other hexose phosphates were tested and were not effective . In the presence of amounts of inhibitor sufficient to produce complete inhibition of hexokinase D, the concentration of fructose 2,6-bisphosphate required to produce 50% inhibition was about 0.5 microM . The inhibitor was unstable and was stabilized by the presence of fructose 2,6-bisphosphate. Eur J Clin Microbiol, 1987 Aug, 6(4), 392 - 4 Evaluation of a new slide latex agglutination test for diagnosis of vaginal candidosis; Hopwood V et al.; A new commercial slide latex particle agglutination test for rapid (2 min) diagnosis of vaginal candidosis was evaluated and compared with conventional methods . Of the 263 women studied, 63 (23.9%) had yeasts in the vagina . Clinical signs of vulvitis or vaginitis were seen in 23 women (8.8%) and 40 (15.2%) were harbouring yeasts without clinical signs . Yeast counts were generally higher in women with clinical signs of vaginal candidosis than in those without . The test was positive in 15 of the 23 women (65.2%) with clinical signs, the incidence of a positive test increasing in direct proportion to the amount of yeasts isolated . The test's sensitivity, specificity and predictive values were comparable to those of microscopy and culture . Being both rapid and simple to perform, this new test offers a useful alternative to conventional methods for the diagnosis of vaginal candidosis. Mycopathologia, 1987 Aug, 99(2), 129 - 31 Studies on the isolation, growth and maintenance of Malassezia pachydermatis; Lorenzini R et al.; Results related to the isolation, cultivation, culture and maintenance of the opportunistic pathogen Malassezia pachydermatis are reported . A dextrose nutrient medium with 1.5% yeast extract turned out to be the most favourable medium for its development . It permitted identification in 24 hours and maintenance of isolates for three months without subculturing . Addition of Tween 80 (1%) significantly enhanced the isolation of this yeast from clinical materials. J Parasitol, 1987 Aug, 73(4), 792 - 6 In vitro cultivation of Paragonimus miyazakii and P . ohirai; Hata H et al.; Excysted metacercariae of Paragonimus miyazakii and P . ohirai were cultured in various media at 37.5 C in a 5% CO2 atmosphere . Paragonimus miyazakii grew rapidly and showed a well-developed ovary, uterus, and testes at 172 days in NCTC 109 supplemented with 30% rabbit serum, 50% egg yolk-109, and rabbit red blood cells (RBC's) . However, none of the worms formed yolk or eggs in these cultures . On the other hand, P . ohirai grew to the adult stage, in which vitellaria and imperfect ova were formed, in NCTC 109 supplemented with 30% dog serum, 10% yeast extract Earle's solution (YLE), and dog RBC's at 252 days . The maximum body length of these worms measured 7.0 mm (mean 5.5 mm) at 252 days . The dog RBC's were an essential ingredient of the culture medium for the development of P . ohirai . Additions of liver concentrate, chick embryo extract (CEE), and egg yolk-109 in the medium did not provide any additional benefits for the development of worms . Using this supplemented medium, adult worms of P . ohirai removed from rats were maintained in vitro to examine their ability to lay eggs . Egg laying occurred during the first 10-13 days for worms that survived more than 60 days . The number of eggs deposited in this medium was about 2 times that found when Hanks' BSS and NCTC 109 were used. J Bacteriol, 1987 Aug, 169(8), 3525 - 30 Purification and properties of two isozymes of pyruvate kinase from Mucor racemosus; Hohn TM et al.; The dimorphic phycomycete Mucor racemosus was found to contain up to five electrophoretic forms of pyruvate kinase (ATP: pyruvate 2-O-phosphotransferase, EC 2.7.1.40) depending on growth conditions . M . racemosus hyphal cells grown on glutamic acid as the carbon source contained only the fastest electrophoretic form, designated PK1, while yeast cells grown on glucose contained only the slowest electrophoretic form, PK5 . Intermediate electrophoretic forms PK2, PK3, and PK4 as well as PK1 and PK5 were found in hyphal cells grown on media containing fructose or cellibiose . All five electrophoretic forms had molecular weights of ca . 230,000 as determined from plots of log Rm versus acrylamide gel concentration . Both PK1 and PK5 were purified to homogeneity and determined to be homotetramers, with subunit molecular weights of 54,000 and 58,100, respectively . The amino acid content of PK1 and PK5 was determined and found to be similar but not identical . Analysis of limited tryptic digests and cyanogen bromide cleavage fragments of PK1 and PK5 indicate that the subunits of the two isozymes are significantly different. Immunol Cell Biol, 1987 Aug, 65 ( Pt 4), 329 - 35 Enhancement of neutrophil-mediated phagocytosis by human granulocyte-macrophage colony-stimulating factor demonstrated using a novel mathematical model; Williamson DJ et al.; A mathematical model is presented which may be applied to describe and analyse data from microscopic phagocytosis assays . The method has been used to investigate the phagocytosis of opsonized yeast by peripheral blood neutrophils treated with purified recombinant human granulocyte-macrophage colony-stimulating factor (rH GM-CSF) in vitro . Under limiting conditions of serum opsonization, rH GM-CSF decreased the proportion of non-phagocytic cells and increased the mean number of ingested yeast per cell . Stimulation of phagocytosis was dose-dependent and occurred with concentrations of rH GM-CSF in the range 10-320 units/ml . The effect was dependent on a heat-labile component in serum and was not attributable to endotoxin contamination of the preparation. J Exp Med, 1987 Aug 1, 166(2), 476 - 88 Characterization of the human B cell stimulatory factor 1 receptor; Park LS et al.; 125I-labeled recombinant human B cell stimulatory factor 1 (BSF-1) was used to characterize receptors specific for this lymphokine on in vitro cell lines representing human B, T, and hematopoietic lineages, as well as on adherent cell lines of epithelial and endothelial origin, and on primary human gingival fibroblasts . BSF-1 binding was extremely rapid and saturable at both 4 and 37 degrees C, with a slow dissociation rate . On all human cell types examined, BSF-1 bound to a single class of high-affinity receptor (less than 3,000 receptors per cell) with a Ka of 0.5-1.0 X 10(10)/M . Human BSF-1 also bound to cell lines of simian but not murine origin . Comparison of kinetic characteristics obtained with a yeast-derived hyperglycosylated form of BSF-1 (Mr 60,000) and N-glycanase-treated, sugar-free BSF-1 (Mr 15,000) showed no significant differences . Among a panel of lymphokines and growth hormones, only unlabeled human BSF-1 was able to compete for the binding of 125I-labeled human BSF-1 . Affinity crosslinking experiments resulted in the identification on both Raji cells and on primary human gingival fibroblasts of a receptor subunit with an average Mr of 139,000 . These studies show that the BSF-1 receptor on human cells has an extremely broad cellular distribution, while further supporting the notion that the ability of BSF-1 to mediate a spectrum of biological activities cannot be accounted for by overt differences in the receptor for this lymphokine on different cell lineages. Exp Appl Acarol, 1987 Aug, 3(3), 191 - 200 Differences in development time, mortality and water loss between eggs from laboratory and wild populations of Dermatophagoides pteronyssinus (Trouessart, 1897) (Acari: Pyroglyphidae); Colloff MJ; A total of four microcultures of adults of Dermatophagoides pteronyssinus, two each from laboratory and wild populations, were fed on separate diets of house dust and yeast granules . A total of 35 eggs of known age from each of the four microcultures were harvested and incubated at 15 degrees C, 60% RH for 16 h/day and 30 degrees C, 75% RH for 8 h/day to simulate diurnal microclimatic fluctuations in a bed . Eggs from females fed on yeast were larger and underwent more rapid rates of water loss, estimated by measurements of reduction in visible surface area (VSA), than eggs from females fed on house dust . There were no significant differences in mean egg development time between the four microcultures (range 6.0-6.88 days) . Mortality of the eggs was as follows: from laboratory females fed on yeast, 31.4%; laboratory females fed on house dust, 11.5%; wild females fed on yeast, 2.9%; wild females fed on house dust, 0% . Thus diet and egg size at oviposition had no effect on mortality . Since the microclimates at which eggs from both populations were oviposited and incubated were identical, it is hypothesized that mortality was higher in eggs from laboratory cultures because the mites had become acclimated to the optimal conditions at which they had been kept and were less able to withstand the diurnal fluctuations in microclimate, similar to those imposed upon wild mites in their natural habitats. J Cell Biol, 1987 Aug, 105(2), 991 - 7 Multiple carbohydrate receptors on lymphocytes revealed by adhesion to immobilized polysaccharides; Brandley BK et al.; Phosphomannan polysaccharides and fucoidan, a polymer of fucose 4-sulfate, have been demonstrated to inhibit adhesion of lymphocytes to tissue sections that contain high endothelial venules (Stoolman, L . M., T . S . Tenforde, and S . D . Rosen, 1984, J . Cell Biol., 99:1535-1540) . We have investigated the potential cell surface carbohydrate receptors involved by quantitating adhesion of rat cervical lymph node lymphocytes to purified polysaccharides immobilized on otherwise inert polyacrylamide gels . One-sixth of the lymphocytes adhered specifically to surfaces derivatized with PPME (a phosphomannan polysaccharide prepared from Hansenula holstii yeast), whereas up to half of the cells adhered to surfaces derivatized with fucoidan . Several lines of evidence demonstrated that two distinct receptors were involved . Adhesion to PPME-derivatized gels was labile at 37 degrees C (decreasing to background levels within 120 min) whereas adhesion to fucoidan-derivatized gels was stable . Soluble PPME and other phosphomannans blocked adhesion only to PPME-derivatized gels; fucoidan and a structurally related fucan blocked adhesion to fucoidan-derivatized gels . Other highly charged anionic polysaccharides, such as heparin, did not block adhesion to either polysaccharide-derivatized gel . Adhesion to PPME-derivatized gels was dependent on divalent cations, whereas that to fucoidan-derivatized gels was not . The PPME-adherent lymphocytes were shown to be a subpopulation of the fucoidan-adhesive lymphocytes which contained both saccharide receptors . These data reveal that at least two distinct carbohydrate receptors can be found on peripheral lymphocytes. EMBO J, 1987 Aug, 6(8), 2425 - 31 A short amino-terminal segment of microsomal cytochrome P-450 functions both as an insertion signal and as a stop-transfer sequence; Sakaguchi M et al.; Co-translational insertion of liver microsomal cytochrome P-450 into the endoplasmic reticulum membrane is mediated by the signal recognition particle (SRP) and the presence in the cytochrome molecule of a signal sequence that can be recognized by SRP has been postulated . To locate this signal sequence, six hybrid cDNAs were constructed in which various segments of a cDNA for a rabbit liver cytochrome P-450 are fused with a cDNA or its fragment encoding yeast porin (an outer mitochondrial membrane protein) or with a cDNA for pre-interleukin 2 (a secretory protein) from which the 5'-terminal portion encoding most of its signal sequence had been removed . These hybrid cDNAs were inserted into an SP-6 transcription vector and transcribed in vitro . The mRNAs thus synthesized were translated in a cell-free system in the presence of rough microsomes . It was thus found that only those chimeric proteins containing (at their amino-terminal end) the amino-terminal cytochrome P-450 segments consisting of greater than or equal to 29 amino acid residues were co-translationally inserted into the membrane in an SRP-dependent fashion . These proteins were, however, neither processed nor translocated across the membrane . These findings, coupled with the observation that the major portion of these proteins, when inserted into the membrane, was degraded by trypsin, led to the conclusion that a short amino-terminal segment (less than 29 residues) of the cytochrome P-450 functions not only as an insertion signal but also as a stop-transfer sequence . This segment is, therefore, similar to the internal signal of type II plasma membrane proteins, but differs from the latter in the topogenic function. Proc Natl Acad Sci U S A, 1987 Aug, 84(16), 5720 - 4 Identification of an acetylation site of Chlamydomonas alpha-tubulin; LeDizet M et al.; An acetylation site of Chlamydomonas axonemal alpha-tubulins was identified near, or within, the binding site of 6-11B-1, a monoclonal antibody specific for posttranslationally acetylated alpha-tubulins . In a first approach, axonemal proteins were hydrolyzed by formic acid, cyanogen bromide, or chymotrypsin and analyzed with immunoblots . The smallest alpha-tubulin peptide retained on nitrocellulose and containing antibody-binding site(s) was found to span amino acids 37-138 (alpha 37-138) . A smaller antibody-binding peptide, identified as alpha 25-50, was obtained by complete digestion of alpha-tubulin with chymotrypsin . This fragment was purified by reversed-phase HPLC and assayed by its ability to bind 6-11B-1 in solution . Determination of the amino acid sequences of alpha 37-138 and alpha 25-50 showed that residue 40 in axonemal alpha-tubulin is epsilon N-acetyllysine . A sequence very similar to Chlamydomonas alpha 25-50 is found in the majority of alpha-tubulins analyzed so far . However, the corresponding region is markedly divergent in some alpha-tubulin isoforms from chicken, Drosophila, and yeast. Clin Chim Acta, 1987 Jul 30, 167(1), 59 - 65 Biosensor for lactate determination in biological fluids . 2 . Interference studies; Racek J et al.; The selectivity of a yeast lactate biosensor with immobilized cells of aerobic yeast Hansenula anomala was studied . Reducing substances potentially present in blood plasma influenced both enzyme and yeast biosensors in the same way; the highest positive error was observed in the case of uric acid . With respect to the metabolic activity of the yeast cells the biosensor was absolutely specific for lactate during the first two weeks; later on the biosensor responded slightly to some other metabolites, especially some sugars and amino acids . Glucose could cause the highest degree of interference, its effect was however completely eliminated by adding sodium fluoride to the reaction solution . The concentration of other metabolites present in blood plasma is not great enough to call a significant positive error . The results thus support the general use of the yeast lactate biosensor for lactate determination in biological material. Biochemistry, 1987 Jul 14, 26(14), 4554 - 8 Sequence dependence for the energetics of dangling ends and terminal base pairs in ribonucleic acid; Sugimoto N et al.; Stability increments of terminal unpaired nucleotides (dangling ends) and terminal base pairs on the core helixes AUGCAU and UGCGCA are reported . Enthalpy, entropy, and free energy changes of helix formation were measured spectrophotometrically for 18 oligoribonucleotides containing the core sequences . The results indicate 3' dangling purines add more stability than 3' dangling pyrimidines . In most cases, the additional stability from a 3' dangling end on an AU base pair is less than that on a GC base pair {Freier, S.M., Burger, B.J., Alkema, D., Neilson, T., & Turner, D.H . (1985) Biochemistry 22, 6198-6206} . The sequence dependence provides a test for the importance of dangling ends for various RNA interactions . Correlations are suggested with codon context effects and with the three-dimensional structure of yeast phenylalanine transfer RNA . In the latter case, all terminal unpaired nucleotides having stability increments more favorable than -1 kcal/mol are stacked on the adjacent base pair . All terminal unpaired nucleotides having stability increments less favorable than -0.3 kcal/mol are not stacked on the adjacent base pair . In several cases, this lack of stacking is associated with a turn in the sugar-phosphate backbone . This suggests stability increments measured on oligoribonucleotides may be useful for predicting tertiary structure in large RNA molecules . Comparison of the stability increments for terminal dangling ends and base pairs, and of terminal GC and AU base pairs, indicates the free energy increment associated with forming a hydrogen bond can be about -1 kcal/mol of hydrogen bond. Arch Tierernahr, 1987 Jul-Aug, 37(7-8), 551 - 7 {The dose dependence of 15N-incorporation in organ proteins of newborn rats after pulse labeling with different tracers}; Wutzke KD et al.; A short-chain 15N-peptide mixture characterized by an average chain length of 2.3 was obtained when 15N-labeled yeast protein has hydrolyzed enzymatically by thermitase from Thermoactinomyces vulgaris . Fifteen newborn Wistar-rats were given a single pulse of {15N}glycine . {15N}H4Cl and {15N}yeast protein-thermitasehydrolysate (YPTH) in a dosage of 50 mg 15N excess kg-1 by gastric tube . In comparison with {15N}glycine the 15N-incorporation rates of brain, muscle and liver were approximately 150% higher after {15N}YPTH-application . Uniform labeling, high 15N-enrichment, almost complete absorption, avoidance of imbalances and the low price make this tracer substance superior to other tracers conventionally used for organ labeling. Yale J Biol Med, 1987 Jul-Aug, 60(4), 333 - 9 Hepatitis B vaccine: prospects for duration of immunity; Krugman S et al.; The duration of hepatitis B vaccine-induced immunity was studied in a group of 54 seronegative health professionals who received plasma-derived hepatitis B vaccine (Merck's Heptavax) in 1978 and 1979 . Five to seven years later, 52 vaccinees received a booster dose of yeast recombinant hepatitis B vaccine (Merck's Recombivax) . Of 54 vaccinees, 47 (87 percent) had a favorable anti-HBs response (greater than 10 S/N RIA units) and 7 (13 percent) had low (2.1-10 S/N) or undetectable levels (less than 2.1 S/N) one year after primary immunization . After five to seven years, the anti-HBs values had declined to undetectable levels in 25 percent and to low levels in 23 percent . A booster dose of vaccine induced an anamnestic response in 90 percent of vaccinees by two weeks . The results of this study indicate that persons who respond favorably to primary immunization may be protected for at least seven years. Prikl Biokhim Mikrobiol, 1987 Jul-Aug, 23(4), 522 - 6 {A method of quantitative determination of delta 5,7-sterols in unsaponifiable lipid fractions}; Zhakovskaia ZA et al.; The technique is offer for rapid quantitative definition of sterols having a system of delta 5,7-conjugated double bonds which conditions biological activity of D vitamin--the result of photoisomerisation of the sterols . The technique bases on the analysis of UV absorption spectra of unsaponifiable lipid fraction of several kinds of mutant yeast strains and the model mixtures of sterols . The results obtained concerning the estimation of delta 5,7-sterols containing in total fraction are confirmed by gas-liquid-chromatography and mass-spectrometry data. J Neurol Sci, 1987 Jul, 79(3), 287 - 300 Fetal brain development in response to iodine deficiency in a primate model (Callithrix jacchus jacchus); Mano MT et al.; The common cotton-eared marmoset (Callithrix jacchus jacchus) has been used for the first time as a primate model to study the effects of dietary iodine deficiency on fetal brain development . Paired male and female marmosets were fed a low-iodine diet of maize, peas, meat meal, Torula yeast, maize oil and added vitamins, minerals and amino acids for 6 months before mating . Offspring from first and second pregnancies were compared with offspring from control marmosets fed the same diet but supplemented with iodine . Severe iodine deficiency in the fetus at birth was evident by reduced plasma thyroxine levels, increased plasma thyroid stimulating hormone levels, increased thyroid weight and reduced thyroid iodine content . Thyroid histology revealed hyperplasia, hypertrophy and absence of colloid material in the follicles . Iodine deficiency caused a reduction in the weight of the fetal brain and in particular the cerebellum . Brain cell number was reduced in the cerebellum and brainstem but cell size was reduced in the cerebral hemispheres . Histology of the brain revealed morphological changes in the cerebellum and cerebral hemispheres . In the-cerebellum there was: an increase in the thickness of the external germinal layer indicative of impaired cell acquisition; a decrease in total area; a decrease in molecular layer area; and an increase in Purkinje cell (Pc) linear density due to a reduction in the length of the Pc line . The decrease in molecular layer area and increase in Pc linear density imply diminished ascending and lateral extension of Pc dendrites . Changes in the cerebral hemispheres consisted of an increase in the density of neuronal cell bodies in the granular band and a decrease in synaptic counts in the layer between the pia mater and supragranular band of the visual cortex . Offspring from second pregnancies compared to those from first pregnancies were more severely affected and associated with lower plasma levels of maternal and fetal thyroxine . These findings indicate the importance of maternal and fetal thyroid function in relation to fetal brain development in the primate. Hum Pathol, 1987 Jul, 18(7), 740 - 5 Scanning electron microscopy of Malassezia furfur attachment to Broviac catheters; Powell DA et al.; Malassezia furfur has been increasingly associated with Broviac-catheter-related sepsis in infants receiving fat emulsions for parenteral alimentation . We examined by scanning electron microscopy the appearance of M . furfur attached to Broviac catheter segments mock-infected in vitro and to Broviac catheters removed from two infants with catheter-related sepsis . In vitro attachment occurred equally on external and internal surfaces of the catheters . Although some organisms were attached next to surface defects in the catheters, we could not determine if such defects were preferential sites of attachment . In the patient catheters, a dense coating of yeast cells was found adhering to the luminal surface, most abundantly near the tip . No organisms were seen on the external surface of the catheters . These findings show the need to examine the mechanisms of intraluminal catheter colonization in order to understand better the pathogenesis of M . furfur infections. Indian J Lepr, 1987 Jul-Sep, 59(3), 247 - 62 Repeated isolation of nocardia like organisms from multibacillary cases of leprosy; Chakrabarty AN et al.; Nocardia like organisms were isolated from all the 22 multibacillary cases of leprosy, on minimal media consisting of only mineral salts and supplemented with simple C-sources (e.g., liquid paraffin, tetradecane etc.) and N-sources (e.g . ammonium salts, urea, asparagine, gelatin etc.) . Complex organic substances, e.g., xanthine, tyrosine, casein, peptone, meat extract, egg proteins, serum, blood, yeast extract as well as medium 199, did not support the growth of these organisms at all . Paraffin-urea minimal (PUM), paraffin gelatin minimal (PGM) and gelatin minimal medium, as well as the agar slants of these media, selectively allowed good growth of these organisms on which these could be serially propagated continuously, and isolated as pure cultures; these were acid-fast long slender rods which were seen to arise directly from fragmented or unfragmented long, slender hyphae, forming at places mycelial tufts many of which, on ageing, sporulated abundantly . Their acid-fastness was pyridine susceptible and these were DOPA-Oxidase positive; these grew best under reduced 02 tension, at pH 7.0-8.0 and temperature about 28 degrees C . Serologically, these appeared to be sufficiently related to, each other, 2 nocardiae (N . brasiliensis and N . caviae) and some mycobacteria. Rev Infect Dis, 1987 Jul-Aug, 9(4), 743 - 53 Malassezia fungemia in neonates and adults: complication of hyperalimentation; Dankner WM et al.; Until recently, Malassezia furfur was thought to be a pathogen only in tinea versicolor . More recently, this lipophilic yeast has been recovered from sick neonates with catheter-related infections . Malassezia fungemia was studied in seven patients, and the salient features of this infection in patients described in the literature were reviewed . Major risk factors include prolonged hospitalization, the presence of central venous catheters, and the use of intravenous fat emulsions . It is difficult to identify specific manifestations of fungemia in these complex cases occurring in patients with severe underlying disease; however, neonates often present with the signs and symptoms of sepsis and thrombocytopenia, whereas fever may be the only manifestation in adults . Some patients are asymptomatic . When symptoms are present, they resolve upon removal of the colonized catheter . The role of the lipophilic nature of Malassezia in the pathogenesis of infection is apparent from the ability of intravenous fat emulsions to support the growth of the fungus in vitro . A special solid medium that can be used to determine the true prevalence of malassezia fungemia has been devised . M . furfur must be considered in the differential diagnosis of opportunistic infections in patients receiving central hyperalimentation and should be sought by the culture of blood on appropriate medium. Diagn Microbiol Infect Dis, 1987 Jul, 7(3), 161 - 75 Epidemiology, diagnosis, and management of Malassezia furfur systemic infection; Marcon MJ et al.; Malassezia furfur, a normal skin flora yeast, generally associated with very mild superficial skin infections, has become an opportunistic pathogen in patients with deep-line vascular catheters . The use of intravenous fat emulsions appears to have altered the microenvironment of the catheter and allowed colonization and subsequent infection . Dissemination of the organism appears to be limited to the lungs, which may have been previously altered by vascular lipid deposition . Because of the serious underlying disease(s) of patients with M . furfur catheter sepsis, it is difficult to determine the exact role of the organism in the overall status of the patients . At the very least, however, catheter removal or discontinuance of the fat emulsion therapy may be required . Antifungal therapy without either of these two steps has not been shown to be efficacious . Physicians must maintain a high index of suspicion of M . furfur catheter sepsis in the appropriate clinical setting, and laboratory investigators must be prepared to provide appropriate diagnostic methods. Invest Ophthalmol Vis Sci, 1987 Jul, 28(7), 1195 - 9 Immunopathology of acute experimental histoplasmic choroiditis in the primate; Anderson A et al.; The immunopathologic features of experimental acute histoplasmic choroiditis were studied in the nonhuman primate . Using an indirect immunoperoxidase technique, a panel of hybridoma-derived anti-human monoclonal antibodies, recognizing distinct lymphoid cell and macrophage surface antigens, have been adapted for use in the primate system . Twenty-two individual foci of histoplasmic choroiditis from five eyes were studied at time periods from 20 to 60 days post intracarotid injection of yeast phase Histoplasma capsulatum . A mononuclear and granulocytic cell infiltration was seen in all lesions . The predominant cell type was the CAPPEL+ T lymphocyte (suppressor/cytotoxic subset) . Other cell types found in smaller numbers were OKT4+ T cells (helper/inducer subset), OK7+ (peripheral B lymphocytes), IgD+ (mantle B cells) and OKM1+ cells (macrophages and polymorphonuclear leukocytes) . Herein, we present immunopathologic data on the acute phase of experimental ocular histoplasmosis. J Immunol, 1987 Jun 15, 138(12), 4313 - 21 Leukocyte-endothelial cell recognition: evidence of a common molecular mechanism shared by neutrophils, lymphocytes, and other leukocytes; Lewinsohn DM et al.; The interaction of leukocytes with endothelial cells is intrinsic to the process of leukocyte extravasation, whether during the entry of blood polymorphonuclear leukocytes and monocytes into sites of acute and chronic inflammation, or during the homing of lymphocytes to lymphoid organs . A lymphocyte surface glycoprotein, defined by monoclonal antibody MEL-14, has been described that appears to mediate lymphocyte recognition of postcapillary venules in peripheral lymph nodes, and to control the migration of lymphocytes from the blood into these lymphoid organs . We now report that the antigenic determinant recognized by MEL-14 is present at high levels on other leukocytes as well, including neutrophils, monocytes, and eosinophils; and we demonstrate involvement of the MEL-14 antigen in neutrophil-endothelial cell interactions . MEL-14 immunoprecipitates a neutrophil surface protein of Mr approximately 100,000, similar in m.w . to the 80,000 to 90,000 dalton lymphocyte surface MEL-14 antigen, and it blocks the interaction of neutrophils with endothelial cells in an in vitro model of adhesion to postcapillary venules in lymph node frozen sections . Neutrophil binding to lymph node venules is also inhibited by PPME, a mannose-6-phosphate-rich yeast polysaccharide that is thought to mimic the endothelial cell ligand for the MEL-14-defined lymphocyte receptor . Interestingly, neither MEL-14 nor PPME exhibit a major effect on neutrophil binding to postcapillary venules in Peyer's patches, suggesting that as for lymphocytes, the neutrophil MEL-14 antigen is involved in recognition of tissue-specific endothelial determinants . Finally, we show that MEL-14 inhibits the capacity of neutrophils to migrate from the blood into sites of acute inflammation in the skin . These observations lead us to propose that receptors for tissue-specific endothelial determinants are utilized by neutrophils and lymphocytes and probably other leukocytes during the physiologic process of leukocyte extravasation in vivo. J Immunol, 1987 Jun 15, 138(12), 4169 - 74 Identification of a shrimp-derived allergen as tRNA; Nagpal S et al.; During an attempt to isolate shrimp allergens, evidence was obtained that shrimp ribonucleic acid was capable of eliciting a specific IgE response in man and an experimental animal model system . The shrimp ribonucleic acid was extracted from boiled whole shrimp (Peneaus indicus), and was isolated by salt precipitation and sequential chromatography over DEAE-Sephacel and BioGel P-100 . The allergenic material was identified as a ribonucleic acid based on the following criteria: a maximal absorption at 258 nm, failure to stain positively with Coomassie Brilliant Blue on slab gel electrophoresis, positive staining with ethidium bromide, co-migration with yeast tRNA on submerged gel electrophoresis in 1.5% Agarose M, and sensitivity to ribonuclease T2 and 0.3 M NaOH . Treatment with protease did not alter its allergenic activity . The RNA was capable of binding allergen-specific IgE in sera from two shrimp-sensitive patients, as demonstrated by microELISA and solid-phase radioimmunoassay (SPRIA) using antigen-coated nitrocellulose filter paper discs and purified 125I-labeled goat anti-human IgE . RNA isolated from shrimp by a conventional tRNA isolation procedure also had the ability to specifically bind IgE in the sera of shrimp-sensitive patients . IgE antibodies to shrimp RNA did not recognize yeast tRNA or salmon testes DNA, and were not detected in sera of other subjects . The shrimp-derived RNA was further able to induce a reaginic response in mice . A combination of in vitro aminoacylation of shrimp tRNA and SPRIA resulted in the identification of the allergenic tRNA as tRNA(Tyr) and tRNA(Arg) . Thus, shrimp tRNA is capable of inducing a specific IgE response in man. Eur J Biochem, 1987 Jun 15, 165(3), 473 - 81 Eucaryotic primase; Roth YF; Eucaryotic primase, an enzyme that initiates de novo DNA replication, is tightly associated with polymerase alpha or yeast DNA polymerase I . It is probably a heterodimer of 5.6 +/- 0.1 S . The enzyme synthesizes oligoribonucleotides of about eight residues which are always initiated with a purine . In vitro the polymerase-primase complex initiates synthesis and pauses at preferred sites on natural single-stranded templates . The relative concentrations of ATP and GTP present in the reaction medium modulate the frequency of site recognition . Primase is strongly ATP-dependent in the presence of single-stranded DNA and of poly(dT) . It also synthesizes oligo(rG) in the presence of poly(dC) very efficiently. J Biol Chem, 1987 Jun 15, 262(17), 8131 - 7 Complete nucleotide sequence of cDNA and predicted amino acid sequence of rat acyl-CoA oxidase; Miyazawa S et al.; cDNA clones for rat acyl-CoA oxidase were isolated . The 3.8-kilobase mRNA sequence of the enzyme was completely covered by two overlapping clones . The composite cDNA sequence consisted of 3741 bases and contained a 1983-base open reading frame which encodes a polypeptide of 661 amino acid residues . Two species of acyl-CoA oxidase cDNA were identified . They differed in their coding nucleotide sequences, only within a small region . They contained the same number of nucleotides and can be translated in a common reading frame . They are 55% and 50% homologous in the above region at the nucleotide and the amino acid levels, respectively . Both types of cDNA were isolated from a library constructed from mRNA of a single rat, thereby suggesting the occurrence of two species of acyl-CoA oxidase in each rat . The amino terminus of the enzyme was determined to be N-acetylmethionine, which corresponds to the initiator methionine, thus confirming the absence of a terminal presequence . We reported previously that a purified preparation of the enzyme contained three polypeptide components, A, B, and C, and suggested that components B and C are produced by a proteolytic cleavage of component A (Osumi, T., Hashimoto, T., and Ui, N . (1980) J . Biochem . (Tokyo) 87, 1735-1746) . We located components B and C on the amino- and the carboxyl-terminal sides of component A . Possible functional significances of several stretches of amino acids of the enzyme are discussed, based on the sequence comparison data between rat and yeast acyl-CoA oxidases. J Biol Chem, 1987 Jun 5, 262(16), 7451 - 4 Serum lectin with known structure activates complement through the classical pathway; Ikeda K et al.; Serum mannan-binding protein (MBP), a lectin specific for mannose and N-acetylglucosamine, was revealed to activate the complement system as measured by passive hemolysis using sheep erythrocytes coated with yeast mannan . In contrast, rat liver MBP, which shares many properties in common with serum MBP, could not activate complement at all . The activation by serum MBP was inhibited effectively by the presence of haptenic sugars and dependent absolutely upon the presence of C4, indicating that the activation is initiated by the sugar binding activity of MBP and proceeds through the classical pathway . The 25 NH2-terminal amino acid sequence of rat serum MBP determined in this study was completely matched with that of MBP-A deduced from cDNA sequence by Drickamer et al . (Drickamer, K., Dordal, M . S., and Reynolds, L . (1986) J . Biol . Chem . 261, 6878-6887), revealing that MBP-A is in fact identical with serum MBP . On the basis of the knowledge of primary structures and physicochemical properties of rat serum and liver MBPs, a possible mechanism of the complement activation by serum MBP is discussed with reference to close similarity in the gross structures of serum MBP and C1q. Mycopathologia, 1987 Jun, 98(3), 171 - 8 Effect of growth of Candida spp . in the presence of various glucocorticoids on the adherence to human buccal epithelial cells; Ghannoum MA et al.; In vitro culturing of three different yeast species with a number of glucocorticoids altered their adherence ability in two ways: Incubation with dexamethasone and triamcinolone acetonide promoted the adherence in general (the increase in adherence ranged between 17% and 44%), whilst growth in the presence of cortisone acetate or hydrocortisone blocked the adherence (inhibition ranged from 16% to 32%) . No statistical difference in the adherence capabilities of different growth phases of C . albicans noted, and the effects of glucocorticoids persisted irrespective of the phase of growth used . An attempt to explain the differences in adherence of the Candida spp . investigated, in the presence of various steroids, on the basis of variation in their structural configurations and/or steroid-receptor interaction is given. Arch Latinoam Nutr, 1987 Jun, 37(2), 378 - 87 {Supplementation of wheat flour with chickpea (Cicer arietinum) flour . I . Preparation of flours and their properties for bread making}; Figuerola FE et al.; The feasibility of adding chick-pea flour substituting part of wheat flour in yeast-leavened bread-making in order to increase the protein value, was studied . A 70% extraction chick-pea flour of commercial granulometry (150 mu) was prepared . Wheat flours of 74% and 78% extraction were then blended with 5%, 10% and 15% of chick-pea flour . Every flour and blend were subsequently analyzed to determine protein, ash, fiber, fat and maltose content, as well as sedimentation, farinogram and bread-making . Addition of chick-pea flour increased protein, fiber, ash and fat content in the blends, not causing a severe effect on quality, even at the 15% level of substitution . Blends showed an increase in maltose content, W value and bread specific volume . Furthermore, breads prepared were of good quality even without the use of maturing agents. Prostaglandins Leukot Med, 1987 Jun, 28(1), 73 - 93 Development of a system for evaluating 5-lipoxygenase inhibitors using human whole blood; Sweeney FJ et al.; A reliable system for evaluating 5-lipoxygenase (5-LO) pathway inhibitors employing human whole blood stimulated by the calcium ionophore, A-23187, and yeast cell walls (YCW) is described . In developing this system, we have shown that leukotriene B4 (LTB4) and 5-hydroxyeicosatetraenoic acid (5-HETE) can be recovered quantitatively from whole blood, and can be measured with accuracy and a precision (standard deviation) of +/- 12% . Apparent differences in LTB4/5-HETE levels between donors can be minimized by normalizing the LTB4/5-HETE production to neutrophil number . Variability in LTB4/5-HETE production among different donors was reduced by increasing the ionophore concentration . The kinetics of ionophore stimulated product production display a 1-4 min lag which is dependent on ionophore concentration . The lag is removed by pretreatment of blood with 5 micrograms/ml cytochalasin B . Likewise, the kinetics of product formation after stimulation with yeast cell walls demonstrated a lag period, which could be shortened by prior opsonization of the YCW . The amount of LTB4 metabolism to 20-OH-LTB4 and 20-COOH-LTB4 in this system is approximately 20% . Phenidone, nordihydroguaiaretic acid, and nafazatrom, known inhibitors of the 5-LO pathway, display half-maximal inhibition points of 0.4, 1.5, and 9 micrograms/ml, respectively . In summary, we believe that this assay offers a guide for predicting systemic levels of drug needed to be achieved for effective inhibition of cellular LTB4/5-HETE synthesis/release in humans. J Immunol, 1987 Jun 1, 138(11), 3897 - 901 Lysosomal enzyme release from human monocytes by particulate activators is mediated by beta-glucan inhibitable receptors; Janusz MJ et al.; Human peripheral blood monocytes ingest particulate activators and generate leukotrienes via a trypsin-sensitive, beta-glucan-inhibitable receptor . The incubation of monolayers of monocytes with from 4 X 10(5) to 2 X 10(8) zymosan or glucan particles resulted in a dose-dependent release of up to 9% +/- 1.9 and 17.8% +/- 5.3 (mean +/- SD, n = 3) of the lysosomal enzyme, N-acetylglucosaminidase, into the culture medium . Lysosomal enzyme release occurred throughout the 2-hr period studied, with the greatest rate of N-acetyl-glucosaminidase release occurring during the first hour; the presence of 5 micrograms/ml of cytochalasin B accelerated this process when zymosan was the agonist . The preincubation of monocytes with from 0.5 to 500 micrograms/ml of soluble yeast beta-glucan inhibited N-acetylglucosaminidase release by 4 X 10(7) zymosan and glucan particles in a dose-dependent manner, with 50% inhibition occurring with 50 micrograms/ml of soluble yeast beta-glucan (mean +/- SD, n = 3) . Preincubation with as much as 5 mg/ml of yeast mannan had no inhibitory effect on N-acetylglucosaminidase release . The pretreatment for 30 min of monolayers of monocytes with 50 micrograms/ml of affinity-purified trypsin, which selectively inactivates the monocyte-phagocytic response to particulate activators, also fully inhibited lysosomal enzyme release induced by zymosan and glucan particles . The inhibitory effects of a soluble ligand, yeast beta-glucan, and of trypsin pretreatment on lysosomal enzyme release correspond to the inhibitory effect of these agents on monocyte phagocytosis of zymosan and glucan particles and thus indicates ligand specificity for the beta-glucan receptor in the release of stored intracellular mediators. Infect Immun, 1987 Jun, 55(6), 1355 - 8 Morphogenesis and pathogenicity of Histoplasma capsulatum; Medoff G et al.; The sulfhydryl blocking agent p-chloromercuriphenylsulfonic acid (PCMS) irreversibly inhibited the mycelium-to-yeast transitions of two virulent strains of Histoplasma capsulatum, G184A and G222B, when the temperature of incubation was raised to 37 degrees C, and the block persisted even after the cultures were washed free of PCMS . Instead of transforming to yeast cells, PCMS-treated mycelia continued to grow as mycelia at the elevated temperatures . A less virulent strain (Downs) was more temperature sensitive, but it showed a similar irreversible effect at 34 degrees C . Therefore, the mycelium-to-yeast transition of H . capsulatum is not required for the adaptation of mycelia to elevated temperatures but probably results from the temperature-dependent activation of yeast-specific genes . The transition to yeast is inferred to be obligate for pathogenicity in mice because PCMS-treated mycelia failed to cause infection, and no fungi were seen in tissues after PCMS-treated mycelia were injected into mice. Arch Dermatol, 1987 Jun, 123(6), 751 - 6 Glucan-induced keratoderma in acquired immunodeficiency syndrome; Duvic M et al.; Six of 20 patients with acquired immunodeficiency syndrome (AIDS) or AIDS-related complex receiving intravenous infusions of soluble glucan (beta-1-3 polyglucose) developed a keratoderma of the palms and soles . The eruption began during the first two weeks of therapy and resolved two to four weeks after its discontinuation . The eruption was different in appearance from our previously reported keratoderma blennorrhagica in AIDS-associated psoriasis . None of the other 735 patients with AIDS or AIDS-related complex not treated with soluble glucan developed a similar keratoderma . The correlation between receiving glucan and the hyperkeratosis is highly significant . Since glucan is a naturally occurring component of the cell walls of yeast, fungus, and some bacterial organisms, recognition of its ability to induce such a striking reaction pattern may be of general significance and interest, although the reaction itself may be limited to patients with AIDS. J Cell Biol, 1987 Jun, 104(6), 1749 - 54 Differential and sequential delivery of fluorescent lysosomal probes into phagosomes in mouse peritoneal macrophages; Wang YL et al.; It has previously been inferred that the fusion of a macrophage secondary lysosome with a phagosome delivers the entire lysosomal contents uniformly to the phagosome . We found, however, that different fluorescent lysosomal probes can enter phagosomes at remarkably different rates, even though they are initially sequestered together in the same organelles . Thus, sulforhodamine is almost exclusively delivered to yeast-containing phagosomes within 2 h of phagocytosis . But fluoresceinated, high molecular weight dextran accumulates in the same phagosomes only over a period of approximately 24 h . We postulate that the delivery of lysosomal contents may involve an intermittent and incremental process in which individual components can be selectively and sequentially transferred. J Biol Chem, 1987 May 25, 262(15), 7363 - 7 The Neurospora crassa metallothionein gene . Regulation of expression and chromosomal location; Munger K et al.; The promoter region of the Neurospora crassa metallothionein gene contains no sequences which are similar to the mammalian or the yeast metal responsive elements (Munger, K., Germann, U . A., and Lerch, K . (1985) EMBO J . 4, 2665-2668) . We therefore studied the regulation of expression of the N . crassa metallothionein gene in response to different metal ions (Cu2+, Cd2+, Zn2+, Co2+, and Ni2+) by Northern analysis . Only copper led to the induction of metallothionein mRNA . In N . crassa cultures inoculated and grown in copper-supplemented media, metallothionein mRNA appeared during the late logarithmic growth period (about 30 h after inoculation) and was detectable for a time period of more than 30 h . In response to copper shock, however, rapidly increasing amounts of metallothionein mRNA were detected within minutes after copper administration at any time in vegetatively growing mycelia of N . crassa . Maximum levels were detected about 1 h after addition of copper to the medium . The half-life time of the mRNA was estimated as 2.5 h . The amounts of copper metallothionein reach a maximum level at 3 h after induction and thereafter remain constant . The rapid induction by copper ions of metallothionein mRNA and metallothionein together with the remarkable stability of the native protein intracellularly suggest that this protein serves an important homeostatic role in the copper metabolism in this fungus . The structural gene of N . crassa metallothionein has been located on chromosome VI using restriction fragment-length polymorphisms as genetic markers. Biochem J, 1987 May 15, 244(1), 219 - 24 Oxidation of protoporphyrinogen to protoporphyrin, a step in chlorophyll and haem biosynthesis . Purification and partial characterization of the enzyme from barley organelles; Jacobs JM et al.; The protoporphyrinogen-oxidizing enzyme from Triton X-100 extracts of the mitochondrial and etioplast fractions of etiolated barley was purified by using ion-exchange and hydroxyapatite chromatography . The purified enzyme from both organelle fractions exhibited a Km of 5 microM and was labile to mild heat and acidification . The pH optimum (5-6) and the substrate-specificity (mesoporphyrinogen was oxidized as rapidly as protoporphyrinogen) revealed properties very different from the protoporphyrinogen-oxidizing enzyme of rat liver or yeast mitochondria, which is specific for protoporphyrinogen as substrate . The purest fractions showed a polypeptide band corresponding to an Mr of approx . 36,000 on SDS/polyacrylamide-gel electrophoresis . This is the first purification and characterization of the enzyme from a plant, and indicates no readily detectable differences between the enzyme isolated from mitochondrial or etioplast fractions, although only the latter organelle has the capacity for both haem and chlorophyll synthesis. Nucleic Acids Res, 1987 May 11, 15(9), 3891 - 906 Structure and sequence of a UDP glucose pyrophosphorylase gene of Dictyostelium discoideum; Ragheb JA et al.; Cell-cell contact and exogenous cAMP regulate the expression of uridine diphosphoglucose pyrophosphorylase (UDPGP) of Dictyostelium discoideum (B . Haribabu, A . Rajkovic and R . P . Dottin, 1986, Dev . Biol., Vol . 113, 436-442) . cAMP appears to regulate gene expression in Dictyostelium by transmembrane signal transduction (B . Haribabu and R . Dottin, 1986, Mol . Cell . Biol . 6, 2402-2408) . To further characterize the mechanism of action of cAMP on the expression of this gene and the nature of the defects in UDPGP mutants that abort development, we sequenced the cDNA and the genomic DNA, including intervening and flanking sequences . The deduced amino acid sequence predicts a polypeptide of 57,893 d . molecular weight . Three short (100-200 nucleotides) A+T rich introns occur within the coding sequences but only one of them contains a sequence TAACTAAC, similar to the yeast lariat acceptor site . The 5' flanking sequences are also A+T rich and contain an oligo A tract (-14 to -24), a TATA box (-25 to -32), and a short G+C rich region (-63 to -101) which may be a control region . From -196 to -209 is a sequence AAAGTAGTATTCAA which matches in 11 of its 14 nucleotides, a sequence found upstream from the hormonally regulated P-enolypyruvate carboxykinase gene of rat. J Biol Chem, 1987 May 5, 262(13), 6280 - 3 Complete nucleotide sequence of cDNA and deduced amino acid sequence of rat liver arginase; Kawamoto S et al.; Arginase (EC 3.5.3.1) catalyzes the last step of urea synthesis in the liver of ureotelic animals . The nucleotide sequence of rat liver arginase cDNA, which was isolated previously (Kawamoto, S., Amaya, Y., Oda, T., Kuzumi, T., Saheki, T., Kimura, S., and Mori, M . (1986) Biochem . Biophys . Res . Commun . 136, 955-961) was determined . An open reading frame was identified and was found to encode a polypeptide of 323 amino acid residues with a predicted molecular weight of 34,925 . The cDNA included 26 base pairs of 5'-untranslated sequence and 403 base pairs of 3'-untranslated sequence, including 12 base pairs of poly(A) tract . The NH2-terminal amino acid sequence, and the sequences of two internal peptide fragments, determined by amino acid sequencing, were identical to the sequences predicted from the cDNA . Comparison of the deduced amino acid sequence of the rat liver arginase with that of the yeast enzyme revealed a 40% homology. Zentralbl Bakteriol Mikrobiol Hyg {A}, 1987 May, 264(3-4), 379 - 85 Light-microscopical appearance and ultrastructure of Blastocystis hominis, an intestinal parasite of man; Matsumoto Y et al.; A study on Blastocystis hominis was undertaken for the purpose of clarifying the morphology of the organism using the following techniques; Giemsa, Heidenhain Iron Hematoxylin, and DAPI stains, phase-contrast microscopy, and transmission- and scanning-electron microscopy . Microscopic examinations of the lumen fluids aspirated at the endoscopical examination revealed the habitation of B . hominis in the lower ileum and cecum of a patient . When examined light-microscopically, organisms from stool materials, cultures, and aspirated intestinal lumen contents of a patient showed morphological resemblances to each other except for variations in size . Vacuolated cells, which were spherical in shape and characteristically had a large central vacuole and a narrow rim of cytoplasm containing nuclei and some inclusions, were the only form of the organism observed in this study, although the contents of the vacuole notably varied . DAPI stains clearly revealed the nucleus and the possible mitochondrion in the narrow rim of cytoplasm . Phase-contrast microscopy of fresh material prepared with physiological saline was recommended for diagnosis . When examined electron-microscopically, the organisms were coated with a capsule that was composed of fine filamentous materials . All the organisms contained a central vacuole although the contents of it varied considerably . The cytoplasm gave the organism a signet ring appearance and contained cristate mitochondria, a great number of ribosomes, Golgi's apparatuses, cytoplasmic microtubules, and nuclei with a nucleolus . Very few of the ultrastructures are those that would be expected of a yeast . Recent occurrences of B . hominis infection in Kyoto City, Japan, during a two-year period (June 1983 to August 1985) were also reported. Mol Biol (Mosk), 1987 May-Jun, 21(3), 678 - 87 {The loss of CpG dinucleotides from DNA . III . Methylation and evolution of histone genes}; Mazin AL et al.; From nucleotide sequences of more than 70 histones genes in 15 species of eucaryotes the probable frequency was determined for CpG----TpG + CpA substitutions, occurring as a result of deamination of 5-methylcytosine residues in DNA . It was found that histone genes differ in the character of CpG methylation with respect to the species studied and may be divided into three groups differing in the value of CpG suppression . In one of them, M-, CpG dinucleotides must have not been methylated throughout the existence of these genes; in another, M+, nearly every other CpG has undergone transition . In the third group, M +/-, no more than 20% of CpG have steadily undergone methylation (and mutation) . The CpG deficiency in M+ and M +/- histone genes is in general proportional to the level of methylation of total DNA in different species . It has been noted that the genes of different core histones in the same organism are characterized, as a rule, by the same type of CpG methylation and belong to the same group . Genes H1 and H5 show a higher level of CpG suppression and thus have a higher degree of methylation than the genes of core histones from the same organism . The most conserved among the histone genes, those for H3 and H4 in particular, must have not been methylated in the majority of the species studied . The distribution of methylated and non-methylated spacers and coding sequences of histone genes of man, mouse, hen and yeast reveals a mosaic pattern . It has been found that 5'-flanked regions in most cases are methylated more than respective genes, while the G + C content in them is significantly lower, compared with the coding gene sequences . The absence of methylation in the 5'-regulatory regions does not appear to be mandatory for histone genes . It has been established that the genes of the same histones may differ in the level of methylation even in more or less closely related species . Group M- comprises genes of core histones of man, hen, sea urchin, Drosophila, Neurospora and wheat; group M +/- includes analogous genes of mouse, Xenopus, trout and sea urchins . The results obtained testify against the possible universal involvement of methylation in the regulation of histone gene expression. Toxicol Appl Pharmacol, 1987 May, 88(3), 322 - 8 Maternal selenium deficiency enhances the fetolethal toxicity of methyl mercury; Nishikido N et al.; The effect of maternal selenium deficiency on methyl mercury fetotoxicity was examined in the ICR strain of mice . Pregnant mice were fed either selenium-deficient diets based on torula yeast or selenium-supplemented diets which were identical to the former except that 0.1, 0.2, or 0.4 mg of selenium per kilogram of diet was added as sodium selenite . Fetolethality of methyl mercury was exacerbated by maternal selenium deficiency when mothers were administered sc 15, 25, or 35 mumol/kg/day of methylmercuric chloride (MMC) on the 13, 14, and 15th days of pregnancy . One-tenth part per million of selenium in the diet was sufficient to protect the fetuses against MMC fetolethality when dams were administered 25 mumol/kg/day of MMC . Mercury concentrations in maternal and fetal tissues were independent of the dietary selenium level . Selenium concentration and glutathione peroxidase (GSH-Px) activity in maternal tissues were unaffected by MMC administration . In fetal liver, on the other hand, selenium concentration was increased and GSH-Px activity was decreased concurrently by maternal MMC administration in the selenium-supplemented groups . Therefore, as far as GSH-Px activity was concerned, the bioavailability of selenium was markedly decreased in fetal liver by maternal injection of MMC . The increase in selenium content in fetal liver, which was observed only in the selenium-supplemented groups, may play an important role in protection against fetolethal toxicity of MMC. Proc Natl Acad Sci U S A, 1987 May, 84(10), 3166 - 70 Apparent lack of discrimination in the reading of certain codons in Mycoplasma mycoides; Samuelsson T et al.; We report a cluster of four tRNA genes from Mycoplasma mycoides as well as the sequence of the alanine, proline, and valine tRNAs and the serine tRNA reading the UCN codons (where N stands for G, A, C, or U) . This brings the total number of tRNA genes that we have so far characterized in this organism to 14, 6 of which code for tRNAs that read the codons of family boxes . In each of these latter cases, we found only one gene per family box, and the gene sequence contains a thymidine in the position corresponding to the wobble nucleotide, with the exception of the arginine tRNA gene that has an adenosine in this position . Furthermore, all of the tRNA structures reported here have an unsubstituted uridine in the wobble position . These findings are similar to those reported for mitochondria, especially yeast mitochondria, that contain an arginine tRNA with the anticodon ACG . However, the resemblance is not complete since we have demonstrated the presence of two isoacceptor tRNAs for threonine having uridine and adenosine, respectively, in the wobble position . It is suggested that in the M . mycoides at least some of the family codon boxes are read by only one tRNA each, using an unconventional method without discrimination between the nucleotides in the third codon position. Nippon Yakurigaku Zasshi, 1987 May, 89(5), 269 - 77 {Anti-inflammatory, analgesic and anti-pyretic activities of a new anti-inflammatory compound, 2-{4-(3-methyl-2-butenyl)phenyl} propionic acid (TA), in experimental animals}; Higuchi S et al.; Anti-inflammatory, analgesic and anti-pyretic activities of orally administered TA were investigated in experimental animals . Against acetic acid-induced vascular permeability in mice, carrageenin-induced hind paw edema in rats and ultra-violet ray-induced erythema in guinea pigs, TA produced a dose related inhibition at doses of 40-160 mg/kg, 10-40 mg/kg and 10-40 mg/kg, respectively . TA produced no inhibition against histamine-induced vascular permeability even at a dose of 200 mg/kg in rats . Cotton pellet-induced granuloma and adjuvant-induced arthritis in rats were significantly inhibited by repeated administration of TA at a dose of 50 mg/kg/day for 6 days and 25 mg/kg/day for 6 days, respectively . TA showed a dose related analgesic effect at a dose of 50-200 mg/kg in acetic acid writhing, Randall-Selitto and adjuvant arthritic pain methods . A high dose of TA was needed to produce an analgesic effect in the pressure method using mice . TA produced an anti-pyretic effect against the pyrexia induced by yeast in rats . On the other hand, TA showed no effect against normal body temperature in rats . These results suggest that anti-inflammatory, analgesic and anti-pyretic activities of TA are generally a little weaker than those of ibuprofen, and the mode of action of TA is similar to that of a typical acidic non-steroidal anti-inflammatory drug such as ibuprofen, indomethacin or phenylbutazone . The ulcerogenic activity of TA was about 2 and 4 times weaker than that of ibuprofen in rats and mice, respectively . TA showed a protective effect against gastric necrosis induced by HCl.(ABSTRACT TRUNCATED AT 250 WORDS) Proc Natl Acad Sci U S A, 1987 May, 84(9), 2761 - 5 Stimulation of proliferation, differentiation, and function of human cells by primate interleukin 3; Lopez AF et al.; Cloned gibbon interleukin 3 (gIL-3) was found to stimulate the proliferation and differentiation of human bone marrow cells to produce day-14 granulocyte, macrophage, granulocyte-macrophage, and eosinophil colonies in semisolid agar . In the presence of normal human plasma, gIL-3 stimulated megakaryocytes . In methylcellulose cultures, it stimulated erythroid colonies in the presence, but not in the absence, of erythropoietin . When mature human leukocytes were used, gIL-3 stimulated the function of purified mature eosinophils as measured by the capacity to kill antibody-coated target cells, to produce superoxide anions, and to phagocytize opsonized yeast particles in a manner similar to recombinant human granulocyte-macrophage colony-stimulating factor . In contrast, gIL-3 did not significantly stimulate any of the neutrophil functions tested, whereas human recombinant granulocyte-macrophage colony-stimulating factor was active in these assays . Among cytokines that are active on human hematopoietic cells, gIL-3 thus has a distinct set of functions and may predict the range of actions of the human molecule. Acta Cytol, 1987 May-Jun, 31(3), 243 - 8 Cytomorphology of Alternaria in bronchoalveolar lavage specimens; Radio SJ et al.; Review of the bronchoalveolar lavage specimens from 326 patients resulted in the identification of Alternaria in 28 (8.6%) of the specimens . On Papanicolaou-stained Millipore filters, the most common finding was a yellow-brown-pigmented muriform conidium with characteristic transverse and longitudinal septations . Four of the patients had floccose branched and septated hyphae of Alternaria in addition to conidia . Budding yeast or yeast forms were also present in the lavage fluid of 14 of the patients with Alternaria . Two patients had concurrent Pneumocystis carinii pneumonia, and one patient had cytomegalovirus pneumonitis . No patient developed clinical features of systemic Alternaria infection, and autopsy of four patients did not reveal pneumonia . Alternaria conidia in a bronchoalveolar lavage fluid will usually represent laboratory contaminants or nonpathogenic saprophytes, and their significance lies in distinguishing them from other fungi . However, the expanded use of immunosuppressive therapy and the increasing prevalence of acquired immune deficiency syndrome may render such saprophytes clinically important. Mol Cell Biol, 1987 May, 7(5), 1731 - 9 Primary structure of human nuclear ribonucleoprotein particle C proteins: conservation of sequence and domain structures in heterogeneous nuclear RNA, mRNA, and pre-rRNA-binding proteins; Swanson MS et al.; In the eucaryotic nucleus, heterogeneous nuclear RNAs exist in a complex with a specific set of proteins to form heterogeneous nuclear ribonucleoprotein particles (hnRNPs) . The C proteins, C1 and C2, are major constituents of hnRNPs and appear to play a role in RNA splicing as suggested by antibody inhibition and immunodepletion experiments . With the use of a previously described partial cDNA clone as a hybridization probe, full-length cDNAs for the human C proteins were isolated . All of the cDNAs isolated hybridized to two poly(A)+ RNAs of 1.9 and 1.4 kilobases (kb) . DNA sequencing of a cDNA clone for the 1.9-kb mRNA (pHC12) revealed a single open reading frame of 290 amino acids coding for a protein of 31,931 daltons and two polyadenylation signals, AAUAAA, approximately 400 base pairs apart in the 3' untranslated region of the mRNA . DNA sequencing of a clone corresponding to the 1.4-kb mRNA (pHC5) indicated that the sequence of this mRNA is identical to that of the 1.9-kb mRNA up to the first polyadenylation signal which it uses . Both mRNAs therefore have the same coding capacity and are probably transcribed from a single gene . Translation in vitro of the 1.9-kb mRNA selected by hybridization with a 3'-end subfragment of pHC12 demonstrated that it by itself can direct the synthesis of both C1 and C2 . The difference between the C1 and C2 proteins which results in their electrophoretic separation is not known, but most likely one of them is generated from the other posttranslationally . Since several hnRNP proteins appeared by sodium dodecyl sulfate-polyacrylamide gel electrophoresis as multiple antigenically related polypeptides, this raises the possibility that some of these other groups of hnRNP proteins are also each produced from a single mRNA . The predicted amino acid sequence of the protein indicates that it is composed of two distinct domains: an amino terminus that contains what we have recently described as a RNP consensus sequence, which is the putative RNA-binding site, and a carboxy terminus that is very negatively charged, contains no aromatic amino acids or prolines, and contains a putative nucleoside triphosphate-binding fold, as well as a phosphorylation site for casein kinase type II . The RNP consensus sequence was also found in the yeast poly(A)-binding protein (PABP), the heterogeneous nuclear RNA-binding proteins A1 and A2, and the pre-rRNA binding protein C23 . All of these proteins are also composed of at least two distinct domains: an amino terminus, which possesses one or more RNP consensus sequences, and a carboxy terminus, which is unique to each protein, being very acidic in the C proteins and rich in glycine in A1, and C23 and rich in proline in the poly(A)-binding protein . These findings suggest that the amino terminus of these proteins possesses a highly conserved RNA-binding domain, whereas the carboxy terminus contains a region essential to the unique function and interactions of each of the RNA-binding proteins. Mol Cell Biol, 1987 May, 7(5), 1925 - 32 Expression of three genes for elongation factor 1 alpha during morphogenesis of Mucor racemosus; Linz JE et al.; Three genes, TEF-1, -2, and -3, encode elongation factor 1 alpha in Mucor racemosus . Neutral and alkaline S1 nuclease analyses revealed that the genetic organization is unique for each of the genes . The number and size of the intervening sequences vary in these closely related genes, which suggests that complex genetic rearrangements gave rise to the elongation factor 1 alpha gene family . Nucleotide sequence data from restriction fragments isolated from the 5' and 3' ends of TEF-2 and -3 confirmed the presence of a second intervening sequence in these genes . These data along with S1 nuclease mapping revealed a region at the 3' end of the three genes which was predicted to be transcribed but untranslated . Unique oligonucleotides containing 19 bases were synthesized to hybridize to this unique trailer region in the elongation factor 1 alpha transcripts . These oligonucleotides were used as probes in standard Northern analysis of RNA purified from M . racemosus cells of several morphological types . It was determined that all three genes were expressed in the cell morphological types studied . However, the accumulated level of transcript derived from each gene varied considerably, with TEF-1 mRNA present in approximately twofold greater quantity than the TEF-3 transcript and up to sixfold greater quantity than TEF-2 . The level of TEF-1 and -2 mRNA varied little among the cell morphological types studied, whereas TEF-3 mRNA was present in twofold greater quantity in sporangiospores than in either germlings or yeast cells which had been induced to undergo morphogenesis to hyphae . These data suggest that there is differential expression of the genes encoding elongation factor 1 alpha in M . racemosus . At least one gene, TEF-3, shows a morphology-specific pattern of transcript accumulation. Br J Nutr, 1987 May, 57(3), 319 - 29 Relative nutritional availability to rats of selenium in Finnish spring wheat (Triticum aestivum L.) fertilized or sprayed with sodium selenate and in an American winter bread wheat naturally high in Se; Mutanen M et al.; A Finnish national programme to fertilize crops with sodium selenate led us to compare the nutritional availability to rats of selenium in two Finnish spring wheats (Triticum aestivum L.), either fertilized or sprayed with sodium selenate, with that in an American winter bread wheat naturally high in Se . Weanling male rats were given a Se-deficient Torula yeast diet for 4 weeks followed by either continued depletion or repletion for 4 weeks with graded levels of Se as sodium selenite (standard) or wheat (test food) . Plasma and liver Se levels and plasma and liver glutathione peroxidase (EC 1.11.1.9; GSH-Px) activities were used as criteria of body Se status . The availability of Se under these conditions was calculated with the point-slope technique at two dietary levels of Se (Expt 1) and with the slope-ratio method (Expt 2) . In the point-slope assay, the level of dietary Se fed had a considerable effect on the apparent availability values obtained which made interpretation of the results difficult . In the slope-ratio assay, no difference in the availability of Se from the various wheats was observed when plasma or liver Se levels were used as the response criteria . The Se in the fertilized wheat was somewhat more available than that in the sprayed wheat when plasma or liver GSH-Px activities were the response criteria . Overall, availability values (%) derived by averaging all four response criteria were 86, 77 and 73 for the fertilized and sprayed Finnish wheats and the American wheat respectively (sodium selenite 100) . These results show that wheat is a relatively available source of Se to rats regardless of whether its Se content is naturally high or is increased by fertilization or spraying. Anal Biochem, 1987 May 1, 162(2), 443 - 5 The use of N,N,N',N'-tetramethylphenylenediamine to detect peroxidase activity on polyacrylamide electrophoresis gels; Butler MJ et al.; N,N,N',N'-Tetramethylphenylenediamine (TMPD) acts as an effective indicator of peroxidase activity on polyacrylamide electrophoresis gels . The test is easy to perform, rapid, sensitive, and reliable . The procedure produces vivid bright blue bands (Wursters blue) on a clear background . TMPD and Wursters blue did not interfere with a number of other electrophoresis stains subsequently applied . These included total protein staining with Coomassie blue, and a number of pigment producing electrophoresis stains used to investigate melanogenesis-related enzymes in the black yeast Phaeococcomyces sp. Eur J Biochem, 1987 Apr 15, 164(2), 383 - 7 Evidence for the transcriptional control of nitrate reductase in Candida nitratophila from in vitro translation studies; Cannons AC et al.; In vivo labelling and in vitro translation studies were used to study the regulation of the synthesis of nitrate reductase in the yeast Candida nitratophila . These studies showed that synthesis of the enzyme subunit took place when ammonium-grown cells were nitrogen-starved and this was stimulated by subsequent addition of nitrate . Ammonium-grown cultures did not contain mRNA that could be translated into the nitrate reductase subunit in an in vitro system . Nitrate reductase mRNA could be extracted from nitrogen-starved and nitrate cultures . Synthesis of the enzyme is apparently controlled at the level of transcription in this yeast. Eur J Biochem, 1987 Apr 15, 164(2), 329 - 36 Use of a polymer-bound flavin derivative for the rapid regeneration of NAD(P)+ from NAD(P)H in dehydrogenase systems; Montaine F et al.; An aldehyde derivative of riboflavin was covalently attached by reductive alkylation to soluble polycationic supports . The flavopolymers so obtained were stable under operational conditions . The catalytic efficiency towards oxidation of NADH by these flavopolymers was demonstrated, and the kinetic parameters (Km and kcat) revealed an overall catalytic efficiency (kcat/Km) 185-fold greater compared to riboflavin . Various factors affecting the chemical regeneration of NAD+ from NADH such as pH, ionic strength, nature of the buffer etc . were studied . The most interesting result was the highly favourable influence of borate ions which increased the reaction rate by a factor 2-4 compared to the other buffers . The flavopolymers are very effective for in situ recycling of NAD(P)+ . With up to 300-fold NADH----NAD+ conversions for the system using yeast alcohol dehydrogenase and up to 1500-fold NADPH----NADP+ regenerations for the system using glucose-6-phosphate dehydrogenase . These flavopolymers are superior to previous chemical recycling systems. Biochem Biophys Res Commun, 1987 Apr 14, 144(1), 271 - 6 Carrier RNA enhancement of recovery of DNA from dilute solutions; Gallagher ML et al.; A study of the use of carrier RNA to improve precipitation of DNA from dilute solutions was conducted to define the conditions which optimize DNA recovery . Replicate samples containing labeled pBR322 and increasing concentrations of commercially-available Torula yeast RNA were ethanol precipitated at -20 degrees C for 1 h in microfuge tubes obtained from various manufacturers . Nucleic acids were pelleted by centrifugation for either 5 or 30 min, dried and resuspended . Although recovery was not identical in each type of microfuge tube, in all cases the percent recovery increased when carrier was added . In most cases, extending centrifugation to 30 min did not significantly increase recovery . Recovery of unlabeled DNA's of heterogeneous molecular weight and conformation was also enhanced by the addition of carrier RNA . DNA's recovered by this method can be successfully digested with BamHI and ligated with T4 DNA ligase. Nucleic Acids Res, 1987 Apr 10, 15(7), 2837 - 50 tRNA-like properties of tobacco rattle virus RNA; van Belkum A et al.; The 3' terminal forty nucleotides of tobraviral RNAs readily fold into a tertiary structure, resembling that of tymo- and tobamoviral RNAs . The latter RNAs possess a tRNA-like structure at their 3' end that is recognized by a number of tRNA-specific enzymes (Rietveld et al . (1984), EMBO J . 3, 2613-2619) . Characteristic for their aminoacyl acceptor arm is the presence of a so-called pseudoknot which we now also find in a corresponding position at the 3' terminus of TRV RNA2 (PSG strain) . The nucleotide sequences of all tobraviral RNAs analysed so far indicate that they all possess a similar 3' terminal structure . A domain resembling the anticodon arm of canonical tRNA is not readily recognizable . TRV RNA2 can be adenylated with CTP, ATP; tRNA nucleotidyl transferase and ATP . It is unable, however, to accept any of the twenty common amino acids when incubated with ATP and aminoacyl-tRNA synthetases from wheat germ or yeast . We conclude that TRV RNA contains a tRNA-like structure, which, in contrast to the tymo- and tobamoviral tRNA-like structures, cannot be aminoacylated . It is unlikely therefore, that aminoacylation of plant viral RNAs with a tRNA-like structure is a prerequisite for viral RNA replication. Biochemistry, 1987 Apr 7, 26(7), 2060 - 6 Structural and biochemical properties of bidentate tetraaquarhodium(III) complexes of inorganic pyrophosphate and adenosine 5'-diphosphate; Shorter AL et al.; The structural and biochemical properties of the alpha,beta-bidentate tetraaquarhodium(III) complexes of inorganic pyrophosphate {Rh(H2O)4PP} and adenosine diphosphate {Rh(H2O)4ADP} are examined . These Rh(III) complexes are exchange-inert analogues of the corresponding physiologically important MgIIPP and MgIIADP complexes . The crystal structure of {Rh(H2O)4H2P2O7}+Cl- shows that the six-membered chelate ring adopts a twist-boat conformation with an unusually high puckering amplitude of 0.756 (3) A . The Rh coordination distances average 2.02 (1) A, while the bridge P-O bonds are virtually equal in length . All 10 protons of the complex participate in hydrogen bonding . There are two intramolecular hydrogen bonds between the phosphate oxygen atoms and the axially coordinated water molecules . The Rh(H2O)4PP complex was found to be a substrate for yeast inorganic pyrophosphatase, with Ki = 0.063 (7) mM and Vm = 500 (100) min-1 . The two screw sense isomers of Rh(H2O)4ADP were prepared from (Rp)-{alpha-16O,18O}ADP and assigned configuration on the basis of the magnitude of their 31P NMR isotopic chemical shifts . The Rh(H2O)4ADP complex binds a number of kinases as tightly as MgADP . Arginine kinase and creatine kinase were shown to bind the delta Rh(H2O)4ADP isomer 7 and 45 times tighter, respectively, than the lambda isomer . The reactivity of Rh(H2O)4PP with pyrophosphatase is comparable to that of Cr(H2O)4PP, and the binding affinities of the Rh(H2O)4ADP screw sense isomers for kinases are also comparable to those observed for the corresponding Cr(H2O)4ADP screw sense isomers. J Biol Chem, 1987 Apr 5, 262(10), 4708 - 16 Involvement of phosphatidylinositol and insulin in the coordinate regulation of proteoheparan sulfate metabolism and hepatocyte growth; Ishihara M et al.; A rat hepatocyte cell line was cultured in Higuchi's medium with fetal calf serum and insulin and labeled with 35SO2/4- . The cells were treated with a number of ligands to displace the heparan 35SO4 proteoglycan (HSPG) from the pericellular matrix . Maximum release was obtained with D-mannose-6-PO4 (50 mM), D-glucose-6-PO4 (50 mM), myo-inositol-2-PO4 (2-5 mM), myo-inositol hexaphosphate (2-5 mM), and DL-myo-inositol-1-PO4 (1-2 mM) . D-myo-Inositol-1,3,4-(PO4)3 (1 mM) and L-myo-inositol-1-PO4 (2 mM) were intermediate in their ability to release the cell surface HSPG, whereas heparin (2 mg/ml), yeast phosphomannan (4 mg/ml), D-xylose-1-PO4 (50 mM), D-glucose-6-SO4 (50 mM), and myo-inositol hexasulfate (5 mM) were ineffective . When 35SO2/4- was added to cell cultures, the total cell surface HSPG increased linearly, but the percentage of the total cell surface {35SO4}HSPG that was released by myo-inositol-PO4 increased with time during the labeling period, reaching a maximum of 65% after 5 h . When cells were labeled for 12 h without insulin in the medium, the maximum amount of cell surface HSPG that was released by myo-inositol-PO4 was reduced to 30% . However, when cells labeled in the absence of insulin were treated with phosphatidylinositol-specific phospholipase C and then myo-inositol-PO4, the release of the cell surface {35SO4}HSPG was increased to 73% . When the {35SO4}HSPG that was released from the cell surface by treatment with myo-inositol-PO4 was added to cultures of unlabeled hepatocytes, it was taken up very rapidly and a portion of the internalized HSPG was converted to free heparan SO4 chains which appeared in the nucleus . Uptake was Ca2+- and Mg2+-independent . The amount of {35SO4}HSPG taken up was markedly reduced when the myo-inositol-PO4-releasable {35SO4}HSPG was pretreated with trypsin, thermolysin, alkaline borohydride, or alkaline phosphatase . When the cells were grown in inositol-deficient medium or in the presence of myo-inositol-PO4, the amount of heparan SO4 found in the nucleus was markedly reduced, and the cells no longer exhibited contact inhibition . These effects of myo-inositol deficiency on the growth and nuclear heparan SO4 were accentuated by addition of LiCl to the cultures to prevent phosphatidylinositol synthesis from the endogenous myo-inositol-PO4.(ABSTRACT TRUNCATED AT 400 WORDS) Acta Chir Scand, 1987 Apr, 153(4), 279 - 81 Response of human monocyte phagocytosis to FAM (fluorouracil, adriamycin, mitomycin); Athlin L et al.; The immunosuppressive effect of chemotherapy associated with surgery is poorly understood . The effect of combination chemotherapy--fluorouracil, adriamycin and mitomycin (FAM)--on monocyte phagocytosis was tested in patients operated on for gastric cancer . The adherence, engulfment and total phagocytic process were studied with a fluorescence-quenching technique 1 hour, 24 hours and 1 week after 19 FAM treatments in nine patients . The engulfment of yeast cells and the total phagocytosis were significantly impaired 1 hour post-treatment (p less than 0.05 and less than 0.01, respectively) . These functions had normalized after 24 hours, but after 1 week the engulfment step was significantly depressed (p less than 0.05) . Significantly increased adherence of yeast cells (p less than 0.05) was found at the same time . The initial, transient depression of monocyte phagocytosis probably was related to inactivation of cell-surface receptors . The late (1 week) depression probably was due to bone-marrow toxicity . This functional impairment of the blood monocytes was not reflected as clinical liability to infections. Poult Sci, 1987 Apr, 66(4), 700 - 4 Influence of diet composition on hepatic lipid deposition in two lines of Australorps selected for liver fat differences; Jensen LS; This investigation was conducted to determine if the level of liver fat in Black Australorp laying hens, which had been selected over several generations for high or low liver fat content, would respond to changes in diet composition similar to the response of commercial strains of White Leghorns . In the first experiment, the two selected lines were fed either a corn-soybean diet (CS) or an isonitrogenous and isoenergetic diet containing 4.89% each of fish meal, alfalfa meal, and torula yeast (FAY) for 6 weeks . Liver lipid was significantly higher for the high (19.4%) than for the low line (10.8%), but the FAY diet did not significantly affect this parameter . In a second 6-week experiment, the high and low lines of Black Australorps and unselected commercial White Leghorn hens were fed either the CS diet or one containing 14.67% distillers dried grains with solubles (DDGS) . The DDGS significantly reduced both liver lipid and relative liver weight in Leghorns but not in the low and high liver fat Black Australorp lines . These results indicate that the mechanism of hepatic lipid accumulation in the genetically selected birds is different from that caused by feeding Leghorn hens a simplified CS diet and that the high liver fat line would not be useful in delineating nutritional effects on hepatic lipid accumulation. Ann Rheum Dis, 1987 Apr, 46(4), 302 - 6 Polymorphonuclear neutrophil function in systemic sclerosis; Czirjak L et al.; In vitro functions of polymorphonuclear (PMN) neutrophils were studied in 20 patients with progressive systemic sclerosis (PSS) . An increase in the basal chemiluminescence (CL) activity of peripheral blood PMNs was found, suggesting that these cells had been preactivated in vivo . Patients with more extensive skin disease or signs of disease progression tended to have higher basal CL values . Active oxygen products during the respiratory burst may increase the extent of inflammatory and fibrotic processes and could be involved in the endothelial injury in PSS . The stimulatory capacity of CL response was normal in our study . No alterations were found in the opsonised yeast phagocytic activity of granulocytes when compared with control values . The binding of erythrocyte-antibody particles was found also to be normal . A depressed chemotactic activity of PMN cells against zymosan activated serum was also shown . The cause of the decreased chemotaxis of PMNs remains to be elucidated. J Nutr, 1987 Apr, 117(4), 732 - 8 The effect of progressive selenium deficiency on anti-glutathione peroxidase antibody reactive protein in rat liver; Knight SA et al.; Liver glutathione peroxidase (GSH-Px, EC 1.11.1.9) activity decreases when weanling rats are fed a Se-deficient diet . To determine the effect of dietary Se deficiency on the concentration of the protein portion of GSH-Px, weanling rats were fed a Se-deficient (less than 0.02 ppm Se) or a Se-supplemented (0.2 ppm Se as Na2SeO3) 30% torula yeast-based diet and killed 0, 3, 7, 14, 21 or 28 d later . GSH-Px activity was assayed using H2O2, so only the Se-dependent GSH-Px was measured . Anti-GSH-Px antibodies, produced in a rabbit by three injections of purified rat liver GSH-Px, were used in an enzyme-linked immunosorbent assay for GSH-Px protein . Immunoblotting showed that the antibodies were highly specific for GSH-Px . In Se-supplemented rats, liver GSH-Px activity increased 66% and GSH-Px protein increased 50% over the 28 d . In Se-deficient rats, liver GSH-Px activity decreased exponentially to zero with a half-life of 2.8 d . Liver GSH-Px protein also decreased exponentially, but with a longer half-life of 5.2 d (P less than 0.001), and the anti-GSH-Px antibody-reactive protein did not decrease to zero . This experiment shows that both GSH-Px activity and GSH-Px protein decrease exponentially during progressive dietary Se deficiency . The longer half-life of GSH-Px protein compared with GSH-Px activity suggests that an inactive GSH-Px polypeptide is present in rat liver during the early stages of Se deficiency. J Nutr, 1987 Apr, 117(4), 725 - 31 Relative bioavailability of seleno-compounds in the lactating rat; Smith AM et al.; Bioavailability of the organic forms of selenium (Se), selenomethionine (Se-methionine) and Se-yeast was determined relative to that of an inorganic form, selenite, in the lactating rat . A purified, casein-based diet without added Se was fed to nine groups of rats throughout pregnancy to produce a marginal Se deficiency . During lactation, groups (n = 8) were fed experimental diets containing either 0.1, 0.25 or 0.5 ppm Se as selenite, Se-methionine, or Se-yeast . On d 18 of lactation, tissue Se and glutathione peroxidase (GSH-Px) activities of dams and pups were determined . Based on slope-ratio analyses, the bioavailability of Se-methionine and Se-yeast was greater than that of selenite in both lactating dams and their nursing pups . The greater availability of organic Se to pup tissues may be a direct result of the greater concentration of Se in the milk of dams fed organic Se . A dietary level of 0.25 ppm Se as Se-methionine ensured maximal GSH-Px activity in both dam and pup tissues, but 0.5 ppm Se was necessary when selenite or Se-yeast was fed . These results indicate that, regardless of form, the National Research Council recommendation for growing rats of 0.1 ppm Se is not adequate to replete lactating dams and maintain maximal tissue GSH-Px in nursing pups. J Clin Microbiol, 1987 Apr, 25(4), 605 - 8 Phaeohyphomycotic cyst caused by a recently described species, Phaeoannellomyces elegans; Engleberg NC et al.; An 81-year-old man presented with a chronic, painful nodule on the palmar surface of the left fourth finger . As a former farm worker, the patient acknowledged frequent soil-contaminated wounds of the left hand 4 to 12 years previously, but he denied any recent trauma . The patient's other medical problems included a history of chronic immunoglobulin A gammopathy and a new pleural mass eroding into adjacent ribs on chest X-ray . The finger nodule was excised and consisted of an intact phaeohyphomycotic cyst which yielded growth of a darkly pigmented fungus . At 25 degrees C, the isolate formed annellidic yeast cells having dark-brown walls consistent with the recently described species Phaeoannellomyces elegans . In vitro antifungal susceptibility tests indicated resistance to amphotericin B and variable susceptibility to imidazoles . The lesion was excised, and the patient received no antifungal therapy . After 9 months of follow-up, the fungal infection shows no signs of recurrence. J Biomol Struct Dyn, 1987 Apr, 4(5), 707 - 28 3-D graphics modelling of the tRNA-like 3'-end of turnip yellow mosaic virus RNA: structural and functional implications; Dumas P et al.; The tRNA-like structure of the aminoacylatable 3'-end of turnip yellow mosaic virus (TYMV) RNA was submitted to 3-D graphics modelling . A model of this structure has been inferred previously from both biochemical results and sequence comparisons which presents a new RNA folding feature, the "pseudoknot" . It has been verified that this structure can be constructed without compromising accepted RNA stereochemical rules, namely base stacking and preferential 3'-endo sugar pucker . The model has aided interpretation of previous structural mapping experiments using chemical and enzymatic probes, and new accessibilities of residues could be predicted and tested . Pseudoknots have been considered as potential splice sites because they form antiparallel helical segments in a single RNA molecule . We have examined this possibility with the constructed 3-D model and could verify the hypothesis on a structural basis . The model presents a striking similarity with canonical tRNA and allows a valuable comparison between the protection patterns of yeast tRNA(Val) and tRNA-like viral RNA by cognate yeast valyl-tRNA synthetase against structural probes. J Clin Microbiol, 1987 Apr, 25(4), 675 - 9 Application of DNA typing methods to epidemiology and taxonomy of Candida species; Scherer S et al.; Methods are described for extraction of DNA from the yeast form of Candida spp., followed by digestion and electrophoresis of DNA fragments . The resulting gel patterns (greater than 100 bands) were used to type Candida isolates . Four intense bands identified, three of which are present in each isolate (6 to 7, 3.7 or 4.2, and 2.5 to 3 kilobases), appear to be DNA encoding the rRNA . The methods proved to be both simple and reproducible . The patterns were shown to be stable through several hundred doublings from multiple single colonies . A survey of isolates showed that, on the basis of similarity of gel patterns, several Candida species could be sorted into mutually exclusive groups, and subgroups could be created . Analyses of this survey suggested the possible epidemiologic and taxonomic applications of these methods . DNA typing methods appear to offer important potential advantages over phenotyping methods . The methods provide a base for further epidemiologic studies and for further development of techniques, such as the use of cloned probes for studies of DNA homology. Proc Natl Acad Sci U S A, 1987 Apr, 84(8), 2391 - 5 Transmission of mitochondrial and chloroplast genomes in crosses of Chlamydomonas; Boynton JE et al.; Physical differences between organelle genomes of the interfertile species Chlamydomonas reinhardtii and Chlamydomonas smithii have been used to demonstrate that sexual zygotes transmit chloroplast and mitochondrial DNA from opposite mating types . Processes responsible can be separated functionally and genetically, although both are controlled by mating type . In vegetative diploids, chloroplast and mitochondrial genomes are transmitted biparentally, but a 1-kilobase insert present in the C . smithii mitochondrial genome spreads unidirectionally to all C . reinhardtii genomes in a manner reminiscent of the intron found in the mitochondrial 21S rRNA gene of omega + strains of yeast. Proc R Soc Lond B Biol Sci, 1987 Mar 23, 230(1259), 147 - 61 The Florey lecture, 1986 . Vaccine prevention of virus-induced human cancers; Epstein MA; Carcinogenic viruses have been discovered in numerous animal species over the last 80 years but their role in human cancer has only recently become an important issue . With EB virus involved with endemic Burkitt's lymphoma and undifferentiated nasopharyngeal carcinoma, hepatitis B virus with primary liver cancer, papilloma viruses with carcinoma of the cervix, and T-cell leukaemia virus with adult T leukaemia, 20-25% of all human cancer appears to have a virus component in its causation . By analogy with certain virus-induced animal cancers, vaccine prevention of infection should greatly reduce subsequent tumour development; vaccines against hepatitis B virus are already on trial for this purpose in populations at risk . Experiments are described in which an EB virus subunit vaccine consisting of the virus-determined membrane antigen glycoprotein molecule of molecular mass 340 kDa (MA gp340) has been prepared by two purification methods . Material from one of these has successfully protected cotton-top tamarins against a 100% lymphomagenic dose of challenge virus and investigations are under way to identify an immunogen, based on MA gp340, suitable for use in man . Genetically engineered bacterial, yeast, and mammalian cells expressing the gp340 gene are already available; this gene has also been inserted into vaccinia and varicella virus vectors . Powerful new adjuvants are also considered, together with future strategies for human vaccine studies. J Mol Biol, 1987 Mar 20, 194(2), 345 - 7 Isolation and identification of partial cDNA clones for endoplasmin, the major glycoprotein of mammalian endoplasmic reticulum; Smith MJ et al.; The amino acid sequences of peptides isolated from murine endoplasmin showed significant homology (approximately 50%) with sequences in the heat-shock proteins 90 and 83 of yeast and Drosophila, respectively, indicating that they are related proteins . Mixed oligonucleotide probes, deduced from the peptide sequences, were used to isolate cDNAs from a murine liver cDNA library . DNA sequencing confirmed the presence of a coding sequence for one of the endoplasmin peptides, formally establishing the authenticity of the cDNA . The identity of the murine and hamster endoplasmin sequences suggests a level of sequence conservation associated with proteins that perform a structural role in cells. J Mol Biol, 1987 Mar 20, 194(2), 341 - 4 The glucose-regulated protein grp94 is related to heat shock protein hsp90; Sorger PK et al.; We report the sequence of a cDNA clone that encodes the C-terminal half of the hamster 94 X 10(3) Mr glucose-regulated protein, grp94 . The amino acid sequence of this protein is about 50% homologous to Drosophila hsp83 and yeast hsp90, suggesting that grp94 and hsp90 have similar functional properties . Unlike hsp90, grp94 is associated with the endoplasmic reticulum . It has the same C-terminal tetrapeptide as two other luminal endoplasmic reticulum proteins, grp78 and protein disulphide isomerase . We suggest that this sequence forms part of a signal for retention of proteins in the lumen of the endoplasmic reticulum. Biochim Biophys Acta, 1987 Mar 19, 923(3), 421 - 30 Organothallium(III) reagents for modification of biomacromolecules: irreversible labelling of phosphoglycerate kinase from rabbit muscle; Bunni MA et al.; Organothallium(III) reagents, by analogy with organomercurials, have been found to rapidly label phosphoglycerate kinase from rabbit muscle . By use of a radio-labelled version of p-methylphenylthallium(III) bis-trifluoroacetate (MPT) the inhibition was shown to be irreversible by the criterion of gel filtration desalting . The rate of labelling was shown to depend on the temperature, enzyme and thallium reagent concentrations, and the presence or absence of the various substrates of the enzyme . The structure and oxidation state of the thallium reagent used affected the extent of modification by the compounds MPT, o-carboxyphenylthallium(III) bis-trifluoroacetate, thallic trifluoroacetate and thallous acetate . A number of pieces of evidence implicate cysteine residues in the labelling, including changes in the free thiol titre of the enzyme on thalliation, model studies on the interaction of thiols (e.g . glutathione) with thallium(III) and thallous materials, the lack of inactivation of phosphoglycerate kinase from yeast (which has only one thiol residue distant from the active site), and the partial restoration of enzymic activity by treatment of thalliated enzyme with sulphydryl reducing agents . Substrate protection studies showed that modification of rabbit muscle phosphoglycerate kinase by MPT was fully prevented by 3-phosphoglycerate and partially by MgATP . The latter protected only against the fast phase of thallic modification, the slower phase being unaffected . The presence of MgADP potentiated the labelling by MPT . No evidence of an MgADP-induced conformational change in the enzyme could be obtained from fluorescence or circular dichroic spectroscopies, although changes of the native spectra were noted on thalliation by MPT alone . The cross-linking potential of these arylthallium(III) reagents is discussed along with conformational changes required to trigger the hinge-movement between the N- and C-domains of the protein. Ann Inst Pasteur Microbiol, 1987 Mar-Apr, 138(2), 165 - 76 {Morphogenesis and enzymes of a wild strain and a mutant of Aureobasidium pullulans}; Pasquier-Clouet C et al.; A . pullulans was able to produce several morphological growth forms, including yeast forms (blastospores, swollen cells and chlamydospores) and filamentous forms (true hyphae and pseudomycelium) . Morphological types were dependent on the chemical composition of the media . In nutrient broth medium, culture contained 71% blastospores and 29% pseudomycelium; in yeast extract (YE) medium, 59% blastospores and 41% true mycelium were observed, but not pseudomycelium . Mutagenic treatment by nitrosoguanidine, ethyl methyl sulphonate and orange acridine induced morphological mutant strains . In YE medium, the ratio between cellular and filamentous shapes of a mutant strain was inverted (78% hyphae versus 22% blastospores) compared to the wild type . At the same time, DNase and RNase activities were 5 to 10 times higher. J Ethnopharmacol, 1987 Mar-Apr, 19(2), 193 - 200 Analgesic, antipyretic and ulcerogenic activity of Nyctanthes arbor tristis leaf extract; Saxena RS et al.; The leaves of Nyctanthes arbor tristis, besides being used in the treatment of sciatica and arthritis, are advocated for various kinds of fevers and painful conditions by the Ayurvedic physicians . In the present study, the water-soluble portion of an ethanol extract of the leaves was screened for analgesic, antipyretic and ulcerogenic activities . The extract exhibited significant aspirin-like antinociceptive activity but failed to produce morphine-like analgesia . It was also found to possess antipyretic activity against brewer's yeast-induced pyrexia in rats . The extract also produced gastric ulcers following oral administration for six consecutive days in rats . Results of the present study tend to substantiate the use of this plant in fevers and painful conditions by Ayurvedic physicians. J Ethnopharmacol, 1987 Mar-Apr, 19(2), 185 - 92 Antipyretic studies on some indigenous Pakistani medicinal plants: II; Ikram M et al.; Eight Pakistani medicinal plants were investigated for antipyretic activity in rabbits receiving subcutaneous yeast injections . Hexane- and chloroform-soluble extracts of Aconitum napellus stems, Corchorus depressus whole plant and Gmelina asiatica roots exhibited prominent oral antipyretic activity while insignificant antipyretic effects were found in the hexane- and chloroform-soluble portions of Melia azadirachta seeds, Tinospora cordifolia stems and Vitex trifolia seeds . No antipyretic actions whatsoever were produced by extracts of A . heterophyllum roots and Hedysarum alhagi aerial parts . Toxicity studies revealed no noteworthy toxic or adverse effects for any of the above plant extracts up to the highest oral doses of 1.6 g/kg except in the case of A . napellus. Anal Biochem, 1987 Mar, 161(2), 438 - 41 Effect of the presence of a reversible inhibitor on the time course of slow-binding inhibition; Weiss PM et al.; The half-time for the initial burst seen when a slow-binding inhibitor is present in an enzyme assay decreases from 0.693/k4 to 0.693/(k3 + k4) as the concentration of the slow-binding inhibitor is increased from zero to infinity (k3 and k4 are forward and reverse rate constants for the isomerization causing the slow-binding behavior) . If the inhibitor solution contains a classical reversible inhibitor in addition to the slow-binding one, the half-time decreases from the same limit at zero inhibitor to a level which is higher at infinite inhibitor concentration (k3 is divided by (1 + xKi/Kj), where x is the ratio of classical and slow-binding inhibitor concentrations, and Ki and Kj are their initial inhibition constants before the slow-binding phase) . Thus if one is using a racemic inhibitor, both enantiomers of which inhibit initially but only one of which shows slow-binding behavior, one will not obtain the correct parameters for the pure slow-binding inhibitor . A similar situation would apply if one were using a mixture of inhibitors such as antibiotics, several of which inhibit initially, but only one of which is a slow-binding inhibitor . This theory is illustrated by determining the half-times for the slow-binding inhibition of yeast hexokinase by various levels of TmATP in the presence and absence of HoATP, which shows little slow-binding behavior. Proc Natl Acad Sci U S A, 1987 Mar, 84(5), 1253 - 7 Tunicamycin-resistant Leishmania mexicana amazonensis: expression of virulence associated with an increased activity of N-acetylglucosaminyltransferase and amplification of its presumptive gene; Kink JA et al.; Tunicamycin at 10 micrograms/ml inhibits the growth and infectivity of the parasitic protozoan Leishmania mexicana amazonensis . Tunicamycin-resistant variants of this parasite were produced by gradual acclimatization of cells to increasing concentrations of the drug up to 80 micrograms/ml and a single-step selection of ethyl methanesulfonate-pretreated or differentiating leishmanias with the drug at 10 micrograms/ml . Prolonged exposure to the drug increases stability of drug resistance of those resistant to 10 micrograms/ml . Tunicamycin-resistant cells contain amplified DNA, which hybridizes in proportion to the cells' degree of drug resistance with Alg 7, a cloned DNA probe apparently encoding yeast N-acetylglucosaminyltransferase . This enzyme from all variants remained sensitive to inhibition by tunicamycin, but its specific activity was up to 15-fold higher than that of the wild type . Thus, amplification of the gene encoding this enzyme appears to result in its overproduction in the variants, accounting for their resistance to tunicamycin . The tunicamycin-resistant cells are more virulent to mice than their parental wild type . Thus, leishmanial virulence may be related to amplification or expression of gene(s) encoding enzymes involved in the regulation of N-glycosylation of parasite proteins. J Biol Chem, 1987 Feb 25, 262(6), 2787 - 93 Rat testosterone 7 alpha-hydroxylase . Isolation, sequence, and expression of cDNA and its developmental regulation and induction by 3-methylcholanthrene; Nagata K et al.; A P-450, designated P-450a, with high testosterone 7 alpha-hydroxylase activity was purified from rat liver microsomes . Specific polyclonal antibody against this P-450 was used to screen a lambda gt11 expression cDNA library and a 1687-base pair cDNA was isolated and sequenced . The deduced protein had 492 amino acids, a calculated Mr of 56,016, and it shared 51 and 45% amino acid similarities to P-450e and P-450f, respectively . Regions of similarity were distributed in distinct areas of high and low similarity along the P-450a primary sequence . P-450a cDNA was introduced into yeast cells using the expression vector pAAH5, and the resultant yeast microsomes contained both a protein of identical electrophoretic mobility to that of rat P-450a and testosterone 7 alpha-hydroxylase activity . These results confirm enzyme reconstitution data and antibody inhibition data that P-450a possesses testosterone 7 alpha-hydroxylase activity . The antibody and cDNA probes were used to examine the mechanism of regulation of P-450a by inducers and during development . P-450a and its mRNA were present at low level in newborn rats and increased to maximal level at 1 week of age in both males and females . At age 12 weeks, however, the P-450a level decreased in males but remained elevated in females . Concomitant with the decrease in P-450a in adult males was an increase in level of another immunologically related P-450 . In adult male rats, P-450a was induced almost 5-fold by adminis