Yeasts

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 administration of 3-methylcholanthrene and this induction was the result of an increase in its mRNA . These results establish testosterone 7 alpha-hydroxylase as a member of the P-450e gene family that is developmentally regulated, sex-dependent, and markedly inducible by 3-methylcholanthrene.

Anal Biochem, 1987 Feb 15, 161(1), 20 - 5
A method for assaying orotate phosphoribosyltransferase and measuring phosphoribosylpyrophosphate; Hupe DJ et al.; An orotate phosphoribosyltransferase, OPRTase, assay method which relies upon binding reactant {3H}orotic acid and product {3H}orotidine-5'-monophosphate to polyethyleneimine-impregnated-cellulose resin and collecting on a GFC glass fiber filter is presented . Elution with 2 X 5 ml of 0.1 M sodium chloride in 5 mM ammonium acetate removes all of the orotate and leaves all of the product orotidine monophosphate (OMP) bound so that it may be measured in a scintillation counter . It was found that the addition of 10 microM barbituric acid riboside monophosphate to the reaction mixture prevented the conversion of OMP to UMP and products of UMP . The assay is suitable for measurement of OPRTase activity with purified enzyme or in crude homogenates . A modification of this scheme using commercially available yeast OPRTase and 10 microM of unlabeled OMP provides an assay for phosphoribosylpyrophosphate with a sensitivity such that 10 pmol of PRPP may be measured.

Biochim Biophys Acta, 1987 Feb 11, 890(2), 117 - 26
Dynamic compartmentation of adenine nucleotides in the mitochondrial intermembrane space of rat-heart mitochondria; Gellerich FN et al.; To investigate whether or not the mitochondrial intermembrane space together with the extramitochondrial space form a homogeneous pool for adenine nucleotides, rat-heart mitochondria were studied in reconstituted systems with pyruvate kinase and ADP-producing enzymes with varied localization . In the hexokinase system, ADP is produced extramitochondrially by added yeast hexokinase, whereas in the creatine kinase system mitochondrial creatine kinase is responsible for ADP regeneration in the intermembrane space . The dependence of mitochondrial respiration on the extramitochondrial {ATP}/{ADP} ratio in both systems was investigated experimentally and by means of computer simulation . Near the resting state, higher {ATP}/{ADP} ratios were found in the creatine kinase system than in the hexokinase system at the same rate of respiration . This and the maintaining of a substantial creatine kinase-stimulated respiration in the presence of pyruvate kinase in excess is explained by a two-compartment model considering diffusion limitations of adenine nucleotides . A diffusion rate constant of (8.7 +/- 4.7) 10(4) microliters X mg-1 X min-1 for ADP and ATP was estimated, resulting in rate-dependent concentration differences up to 13.7 microM AdN between the extramitochondrial space and the AdN-translocator at the maximum rate of oxidative phosphorylation of rat-heart mitochondria . The results support the assumption that ADP diffusion towards the AdN-translocator is limited if its extramitochondrial concentration is low, resulting in a dynamic compartmentation of adenine nucleotides in the mitochondrial intermembrane space.

J Biol Chem, 1987 Feb 5, 262(4), 1505 - 9
Squalene synthetase . Solubilization and partial purification of squalene synthetase, copurification of presqualene pyrophosphate and squalene synthetase activities; Kuswik-Rabiega G et al.; Squalene synthetase (farnesyldiphosphate:farnesyldiphosphate farnesyltransferase, EC 2.5.1.21) is an intrinsic microsomal protein that catalyzes the synthesis of squalene from farnesyl pyrophosphate via the intermediate presqualene pyrophosphate . We have solubilized this enzyme from yeast with a mixture of the detergents N-octyl beta-D-glucopyranoside and Lubrol PX . Approximately 50-fold purification of the solubilized activities has been achieved by chromatography on DEAE-cellulose and hydroxylapatite and by isoelectric focusing . The most highly purified preparation has one major band of protein with a molecular weight of 53,000 as estimated by electrophoresis under denaturing conditions . The enzyme may also have been modified by proteolysis during isolation since a 47,000 molecular weight species was also found . The two activities, presqualene pyrophosphate synthetase and squalene synthetase, copurified during isolation.

Eur J Biochem, 1987 Feb 2, 162(3), 493 - 500
The phosphoglycerate kinases from Trypanosoma brucei . A comparison of the glycosomal and the cytosolic isoenzymes and their sensitivity towards suramin; Misset O et al.; Trypanosoma brucei has two phosphoglycerate kinase (PGK) isoenzymes, one is particle-bound and localized in glycosomes while the other is present in the cytosol . The cytosolic isoenzyme (cPGK) was 900-fold purified from cultured procyclic trypanosomes by hydrophobic interaction chromatography on phenyl-Sepharose followed by affinity chromatography on 2',3'-ATP-Sepharose and had a specific activity of 275 units/mg protein . cPGK was compared with the purified glycosomal isoenzyme (gPGK) from bloodstream-form trypanosomes as well as with the commercially available PGKs from yeast, rabbit muscle and Spirulina platensis, a blue-green alga . Like all other PGKs, cPGK was a monomeric protein with a molecular mass of approximately 45 kDa similar to that of the PGKs from other organisms but 2 kDa smaller than that of gPGK . Despite this difference in length and a great difference in isoelectric point, the two trypanosome isoenzymes strongly resembled each other in several respects . The kinetic parameters did not differ significantly from each other or from the PGKs of other organisms . Both trypanosome enzymes resembled the enzyme from S . platensis in that they had an almost absolute requirement for ATP, contrary to the enzymes from yeast and rabbit muscle, which were capable of utilizing GTP and ITP also . This difference in substrate specificity may be related to the amino acid substitutions, Trp 308----His and Ala 306----Glu in the adenine-binding site, which are only found in the two Trypanosoma isoenzymes . Kinetic analysis showed that these substitutions do not prevent binding of the ATP analogues, but probably prevent phosphoryl-group transfer . Both isoenzymes displayed an activity optimum at pH 6.0-9.0 similar to that for the enzyme of yeast . Both gPGK and cPGK were inhibited by the trypanocidal drug Suramin . This inhibition could be described as competitive both with ATP and 3-phosphoglycerate with two inhibitor molecules binding to one molecule of enzyme . The gPGK, however, was much more sensitive (Ki app . = 8.0 microM) to Suramin than either the cPGK (Ki app . = 20 microM) or the enzymes from rabbit muscle (Ki app . = 55 microM), yeast (Ki app . = 167 microM) or S . platensis (Ki app . = 250 microM) . It is suggested that positive charges on the enzyme's surface may play an important role in the potentiation of the binding of the negatively charged Suramin molecule.

Am Rev Respir Dis, 1987 Feb, 135(2), 412 - 7
Growth inhibition of Blastomyces dermatitidis in alveolar and peripheral macrophages from patients with blastomycosis; Bradsher RW et al.; Blastomyces dermatitidis evokes a pyogranulomatous disorder with organisms frequently found inside giant cells . Macrophages from bronchoalveolar lavage fluid and peripheral blood in monolayer cell cultures were challenged with live yeast organisms to examine phagocytosis and intracellular growth . A greater number of macrophages from patients recovering from blastomycosis had phagocytized Blastomyces compared with macrophages from healthy control donors . No differences were detected within the groups between alveolar and peripheral macrophages . Intracellular growth of the fungus was reduced in cultures of both cell types from patients compared with those from control subjects . Supernatants from specific Blastomyces antigen-stimulated lymphocyte cultures were collected, and treatment with the supernatant to control donors' macrophages resulted in increased phagocytosis and inhibition of intracellular growth . Antigen-induced lymphocyte stimulation as a correlate of cellular immunity is qualitatively related to alveolar or peripheral macrophage phagocytosis and growth inhibition of this fungus.

Arch Microbiol, 1987 Feb, 147(1), 64 - 7
Characterization of the amine oxidase involved in the growth of Trichosporon cutaneum X4 on ethylamine as source of carbon, nitrogen and energy; Large PJ et al.; The amine oxidase from Trichosporon cutaneum X4 grown on ethylamine as carbon, nitrogen and energy source was purified to near homogeneity . The purified enzyme showed the highest resistance to heat of any amine oxidase hitherto characterized from a yeast (half-life at 62 degrees C, 14 min) . Measurement of kinetic parameters as a function of carbon chain length showed results typical of a benzylamine oxidase . Both non-denaturing- and sodium dodecyl sulphate-polyacrylamide gel electrophoresis showed multiple bands, and dimethyl suberimidate cross-linking studies revealed that the enzyme consisted of multimers of two polypeptide chains of Mr respectively 19,000 and 26,000 . The smallest structure to show activity probably contained two of each kind of subunit.

Mycopathologia, 1987 Feb, 97(2), 121 - 7
Ultrastructure of Sporothrix schenckii treated with iodine-potassium iodide solution; Hiruma M et al.; The ultrastructural changes produced by iodine-potassium iodide solution on yeast cells of Sporothrix schenckii were investigated by transmission electron microscopy in order to clarify the mechanism of oral potassium iodide therapy for sporotrichosis . Yeast cells were dipped with solutions containing various concentrations of iodine . The rate of germination decreased markedly between the range of iodine concentrations from 0.63 microgram/ml to 5.0 micrograms/ml . No significant ultrastructural changes were seen at the concentration of the iodine of 1.25 microgram/ml (80% germination) or less . In the concentration of 2.5 micrograms/ml (50% germination), normal cells and degenerated cells coexisted . When the cells were treated with 5.0 micrograms of iodine per ml (0% germination) or more, their interior structures were completely destroyed . It is assumed that iodine treatment of the organism causes rapid destruction in the whole cell.

Monatsschr Kinderheilkd, 1987 Feb, 135(2), 99 - 102
{Protein metabolism and lipid balance in dietary treatment of acute enteritis in infants with a defined standardized oligopeptide diet}; Heine W et al.; The advantage of a standardized oligopeptide formula (Peptisorb pad) for dietary management of diarrhea was proved in 10 infants aged 1 to 10 months with body weights between 4000 and 8860 g . The nitrogen balance turned to normal already at day 3 to 4 of the treatment due to the rapid increase of food supply . Whole body protein parameters estimated by 15N yeast protein thermitase hydrolyzate as a tracer substance were already normalized at that time . Protein synthesis amounted to 5.0 +/- 1.5 g/kg/day, protein breakdown to 3.7 +/- 1.5 g/kg/day and net protein gain to 1.3 +/- 0.6 g/kg/day, resp . Reutilisation rate of endogenous nitrogen was found to be 83% . This correlates with the relatively high nitrogen supply of the oligopeptide diet in comparison to mother's milk feeding . Approximately 15% of the administered amount of total nitrogen were excreted in the feces in contrast to only 5.8% of the 15N tracer dose, indicating the higher losses of endogenous nitrogen due to enteritis . The absorption of the medium chain triglycerides from the diet was 97.5% at an average and thus extremely high.

Agents Actions, 1987 Feb, 20(1-2), 50 - 60
The antialgesic drugs: human therapeutic correlates of their potency in laboratory animal models of hyperalgesia; Dubinsky B et al.; This survey discusses the correlation between the oral potency of antialgesic drugs in several pharmacology laboratories and their human oral dose in clinical practice . We also present a brief overview of a few biological assays that have been successfully used to direct the synthesis of newer antialgesic drugs . The laboratory assay that our analysis showed to be most predictive of the clinical analgesic dose is based upon the response of rats to flexion of an arthritic joint . Laboratory ED50 values from the ACh-induced abdominal constriction assay in mice are nearly as predictive while the predictive power of the yeast-induced hyperalgesia assay in rats is somewhat less . Probably because of the small number of experiments, the correlation between the efficacy of these agents in a canine model of synovitis and their clinical doses only reached borderline statistical significance (p = 0.0651) . Regression equations are presented that permit calculations of single clinical analgesic doses from efficacy data in individual tests . Calculation of stepwise multiple regression showed that the clinical dose could be best predicted when efficacy data obtained in the joint flexion assay in rats and the ACh-induced constriction assay in mice are both taken into account . We have concluded that the effective doses are highly predictive of clinical efficacy because these animal assays have been designed to reflect the action of drugs upon prostanoid-induced hyperalgesia.

Mycopathologia, 1987 Feb, 97(2), 101 - 4
Aflatoxin B1 production in orange (Citrus reticulata) juice by isolates of Aspergillus flavus Link; Varma SK et al.; Out of 7 isolates of Aspergillus flavus obtained from rotting orange (Citrus reticulata) fruits, 3 isolates were found to be toxigenic, producing variable amounts of aflatoxin B1 on a semisynthetic liquid medium . The most toxigenic isolates were further evaluated in plain juice and juice supplemented either with 0.05% yeast extract or 0.5% sucrose or both, at three different incubation periods . The maximum yield of aflatoxin B1 (162.5 micrograms/25 ml) was obtained from the juice supplemented with both sucrose and yeast extract within an incubation period of 10 days, whereas a sharp decline in aflatoxin B1 (30 micrograms/25 ml) was observed when incubation was extended beyond 10 days . The addition of yeast extract has a promoting effect on the yield of aflatoxin in comparison to the sucrose.

J Med Vet Mycol, 1987 Feb, 25(1), 47 - 53
Growth of pathogenic Candida isolates anaerobically and under elevated concentrations of CO2 in air; Webster CE et al.; Individual isolates of seven potentially pathogenic yeast species in the genus Candida all grew to some extent in Eagle's minimal essential medium including serum under elevated concentrations of CO2 and in anaerobic gas jars . The C . glabrata and C . tropicalis isolates had the highest anaerobic growth rates and yields, the C . guilliermondii and C . parapsilosis isolates had the lowest, and the C . albicans, C . krusei and C . pseudotropicalis isolates gave intermediate growth rates and yields . The same relative abilities to grow anaerobically were seen when the seven isolates were cultured in liquid and on agar formulations of a peptone-glucose broth and two media containing Yeast Nitrogen Base.

Proc Natl Acad Sci U S A, 1987 Feb, 84(4), 1050 - 4
Production of glycosylated physiologically "normal" human alpha 1-antitrypsin by mouse fibroblasts modified by insertion of a human alpha 1-antitrypsin cDNA using a retroviral vector; Garver RI Jr et al.; Alpha 1-Antitrypsin (alpha 1AT) deficiency is a hereditary disorder characterized by reduced serum levels of alpha 1AT, resulting in destruction of the lower respiratory tract by neutrophil elastase . As an approach to augment alpha 1AT levels in this disorder with physiologically normal human alpha 1AT, we have integrated a full-length normal human alpha 1AT cDNA into the genome of mouse fibroblasts . To accomplish this, the retroviral vector N2 was modified by inserting the simian virus 40 early promoter followed by the alpha 1AT cDNA . Southern analysis demonstrated that the intact cDNA was present in the genome of selected clones of the transfected murine fibroblasts psi 2 and infected NIH 3T3 . The clones produced three mRNA transcripts (5.8, 4.8, and 2.4 kilobases) containing human alpha 1AT sequences, secreted an alpha 1AT molecule recognized by an anti-human alpha 1AT antibody, with the same molecular mass (52 kDa) as normal human alpha 1AT and that complexed with and inhibited human neutrophil elastase . The psi 2 produced alpha 1AT was glycosylated, and when infused intravenously into mice, it had a serum half-life similar to normal alpha 1AT purified from human plasma and markedly longer than that of nonglycosylated human alpha 1AT cDNA-directed yeast-produced alpha 1AT . These studies demonstrate the feasibility of using a retroviral vector to insert the normal human alpha 1AT cDNA into non-alpha 1AT-producing cells, resulting in the synthesis and secretion of physiologically "normal" human alpha 1AT.

J Trauma, 1987 Feb, 27(2), 151 - 4
Oxygen-derived free radical inhibition in the healing of experimental zone-of-stasis burns; Melikian V et al.; When dehydration, infection, and mechanical trauma are prevented, procedures (such as cooling and/or oral antithromboxane) designed to diminish ischemia in experimental zone-of-stasis burns have been associated with no or only minor improvement in wound healing . To test the hypothesis that ongoing skin damage occurring postburn (PB) may in part be due to release of oxygen-derived free radicals during the 16-hour through 4-day PB period of reperfusion in such burns, beginning immediately and for a period of 5 days PB, equal numbers of guinea pigs received: allopurinol 150 mg/kg PO q 6 h vs . placebo, dimethylsulfoxide (DMSO) 75% applied topically q 12 h vs . placebo, or yeast-derived superoxide dismutase coupled with polyethylene glycol (PEG-SOD, Pharmacia) 10,000 U (Fridovich) given IV q 8 h producing a concentration of 16 U/cc of plasma 8 hr after injection vs . placebo . Gross and histologic examination of wounds by a 'blinded' investigator at 1 week and 3 weeks PB revealed no difference between treatment and control groups when rates of re-epithelialization and frequencies of hair-follicle retention were compared . Using the dosages, routes, and model described, treatment of a zone-of-stasis burn with PO allopurinol (a xanthine oxidase inhibitor), topical DMSO (a scavenger of the hydroxyl radical), or IV PEG-SOD (a scavenger of the superoxide radical) during the first 5 days PB was associated with no increase in the rate of re-epithelialization or frequency of hair follicle retention at 1 and 3 weeks PB when compared with controls.

Zhonghua Min Guo Wei Sheng Wu Ji Mian Yi Xue Za Zhi, 1987 Feb, 20(1), 61 - 8
A modified method for rapid mycological diagnosis of Tinea unguium in Taiwan, Republic of China; Jen TM; This is a study to modify and simplify the classical laboratory procedures of mycological diagnosis of Tinea unguium . By means of a tube KOH digestion technique for microscopic examination of nail clippings and a paper disc transfer technique using a more uniform wool-fiber-yeast extract culture fluid as inoculum for all tests necessary for fungal identification, dermatophytes can be identified to a species level . To authenticate the advantage of the etiologic diagnosis of Tinea unguium by mycological studies, the promising result of our previous clinical therapeutic trial with Griseofulvin was presented.

Antibiot Med Biotekhnol, 1987 Feb, 32(2), 144 - 7
{Comparative study of interferon inducers obtained from natural sources}; Timofeev IV et al.; Interferon-inducing and antiviral effects of natural dsRNA preparations of phage phi 6 and yeast cells were studied in the culture of murine cells L-929 and on random bred albino mice . Both the preparations showed interferon inducing activity in the cell culture . However, for realization of their effect modification of the surface cell membrane by polycation exchange resin (DEAE-dextran) was required . The interferon-inducing activity of both of the natural dsRNA in the mice was high . The maximum interferon titers (1280-5120 units/ml) in blood serum were observed 4-6 hours after the inductor intraperitoneal administration . The interferon-inducing activity of the phage dsRNA was high in the cell culture and yeast dsRNA--in mice, respectively . Both the inductors had antiviral activity and protected 15 to 38.9 per cent of the experimental animals from the effect of 100 LD50 of the murine encephalomyocarditis virus and 10 LD50 of the influenza virus A/Aichi 2/68 (H3N2).

Cell, 1987 Jan 16, 48(1), 165 - 73
Proton ionophores prevent assembly of a peroxisomal protein; Bellion E et al.; Peroxisomal matrix proteins are imported into the organelle posttranslationally . Here we report that proton ionophores disrupt the import and assembly of alcohol oxidase, a homo-octameric flavoprotein of the induced peroxisome from the methylotrophic yeast Candida boidinii . When drug is added to cells containing newly synthesized monomeric alcohol oxidase, octamerization fails to occur and a membrane-associated complex is formed instead . The formation of the complex, which appears to face the cytoplasmic side of the membrane, is reversed when drug is removed, leading to the generation of octamer . Surprisingly, when drug is added to cells containing newly assembled octamers, they dissociate into monomers . We suggest that both the complex and the labile octamer are intermediates in the normal assembly pathway of alcohol oxidase and that energy is required for import and maturation of this peroxisomal protein.

Eur J Biochem, 1987 Jan 15, 162(2), 317 - 23
A monoclonal antibody directed against a small subunit of RNA polymerase B blocks the initiation step; Vilamitjana J et al.; Monoclonal antibodies directed against RNA polymerase B of the fungus Podospora were probed for their inhibitory effect on enzyme activity in vitro . From nine antibodies directed against various subunits of the enzyme only two of them totally inhibit RNA polymerase B activity in a non-selective transcription system . Enzymes A and C are not inhibited at all . The two antibodies recognize the same small subunit (B11) of Podospora enzyme B . One of these antibodies has been used as a specific inhibitor to study the function of the B11 subunit . Enzyme activity is partially protected from the inhibition when the enzyme is previously incubated with the template . The antibody does not interfere with enzyme-DNA complex formation but blocks the initiation process in a transcription system using a dinucleotide as initiator . Elongation of RNA chains does not appear to be affected . Also the antibody cross-reacts with yeast and calf thymus RNA polymerase B and has some, but less, effect on initiation by these enzymes . It is suggested the antigenic site corresponding to this antibody may overlap the catalytic initiation site on the native form of RNA polymerase B.

J Biol Chem, 1987 Jan 15, 262(2), 801 - 10
Isolation, complementary DNA sequence, and regulation of rat hepatic lauric acid omega-hydroxylase (cytochrome P-450LA omega) . Identification of a new cytochrome P-450 gene family; Hardwick JP et al.; Lauric acid omega-hydroxylase (cytochrome P-450LA omega) was purified from livers of rats that had been given the hypolipidemic drug clofibrate . Immunoblot analysis with anti-cytochrome P-450LA omega antibody revealed the presence of two clofibrate-induced cytochrome P-450 proteins in both liver and kidney . This antibody was used to screen a lambda gt11 expression library constructed from liver mRNA isolated from rats given clofibrate . The clone, pP-450LA omega, was isolated and found to contain 2062 base pairs with an open reading frame of 509 amino acids (Mr 58,222) . The pP-450LA omega cDNA insert was placed in the yeast expression vector pAAH5, and the resultant plasmid expressed a protein of the same size and activity as cytochrome P-450LA omega . The mechanism of the regulation of the cytochrome P-450LA omega gene by hypolipidemic agents was examined . The rate of cytochrome P-450LA omega gene transcription was increased as early as 1 h after administration of clofibrate, and this increase was followed by an elevation of cytochrome P-450LA omega mRNA, immunochemically detectable protein, and cytochrome P-450LA omega activity . In contrast, no increase in transcriptional activity was detected after clofibrate administration for the cytochrome P-450PCN or P-450b/e genes . The cytochrome P-450LA omega has several structural features in common with other cytochrome P-450s, including a conserved cysteine-containing heme-binding fifth ligand fragment and a hydrophobic amino terminus . Overall, cytochrome P-450LA omega shared less than 35% cDNA nucleotide and amino acid similarity with cytochromes P-450c, P-450d, P-450e, and P-450PCN, indicating that it is a member of a new cytochrome P-450 gene family . Southern blot analysis indicates that this family contains two or three genes.

J Biol Chem, 1987 Jan 5, 262(1), 69 - 74
Properties of anthranilate hydroxylase (deaminating), a flavoprotein from Trichosporon cutaneum; Powlowski JB et al.; Anthranilate hydroxylase was purified from the yeast Trichosporon cutaneum . This enzyme is a simple flavoprotein which apparently does not require any additional cofactor for the conversion of anthranilate to 2,3-dihydroxybenzoate . Anthranilate hydroxylase has Mr of approximately 95,000, with subunit Mr of 50,000; it contains 2 mol of FAD/mol of enzyme . A number of compounds in addition to anthranilate serve as substrates, or effectors, for this enzyme . Oxygen-labeling experiments show that the oxygen atom at the 3-position of the product, 2,3-dihydroxybenzoate, originates from O2, while that at the 2-position is derived from H2O . A mechanism is proposed involving imine formation and hydrolysis during the reaction with the flavin hydroperoxide formed from reduced enzyme flavin and molecular oxygen . This proposal is in accord with the mechanism postulated for other flavoprotein aromatic hydroxylases.

Eur J Biochem, 1987 Jan 2, 162(1), 45 - 8
Nucleotide sequence of cloned cDNA specific for rat ribosomal protein L31; Tanaka T et al.; A cDNA clone, specific for rat ribosomal protein L31, was isolated by hybrid-selected translation from a cDNA library made for 8-9-S poly(A)-rich RNA from regenerating rat liver . The nucleotide sequence of the cDNA was determined . It consists of 25 base pairs from the 5' leading sequence, the entire coding sequence of 378 base pairs and the 3' trailing sequence of 36 base pairs besides the poly(A) tail . The primary structure of protein L31 was deduced from the nucleotide sequence . It consists of 124 amino acids with a relative molecular mass of 14,331 . The calculated amino acid composition is consistent with the reported composition determined for the hydrolysate of L31 . The amino acid sequence showed marked homology with that of yeast ribosomal protein L34.

Ann Nutr Metab, 1987, 31(1), 9 - 17
Dietary ascorbic acid and selenium relationships in the guinea pig; Poovaiah BP et al.; Plasma-, erythrocyte- and liver Se-dependent glutathione peroxidase (GPx) activity, ascorbic acid (AA) concentration, and erythrocyte and liver Se levels were analyzed in groups of guinea pigs fed a Torula yeast-based semipurified diet supplemented with either O (-AA), 200 (+AA), or 400 (++AA) mg of AA/kg of diet and either 0.05 (+Se) or 0.2 (++Se) ppm of Se for 24 days . Plasma, erythrocyte, and liver AA concentration were directly related to the levels of AA fed . There were 21.2 +/- 2.7% and 51.2 +/- 4.5% (p less than 0.0001) increases in erythrocyte and liver Se, respectively, in ++Se guinea pigs compared to +Se guinea pigs . Dietary AA had no effect on erythrocyte or liver Se levels . Plasma, erythrocyte, and liver GPx specific activities were significantly (p less than 0.001) increased in ++Se guinea pigs compared to +Se guinea pigs . In addition, liver, plasma, and erythrocyte GPx activities were directly related to the level of AA fed . These data indicate that dietary AA has no influence on tissue levels of Se but increases plasma, liver, and erythrocyte GPx activities in the guinea pig.

Biokhimiia, 1987 Jan, 52(1), 73 - 81
{Interaction of NAD(H)-dependent dehydrogenases with active dyes and their complexes with transitional metal ions}; Flaksaite SS et al.; The interaction of NAD(H)-dependent dehydrogenases--yeast alcohol dehydrogenase and rabbit muscle lactate dehydrogenase--with reactive dyes produced in the USSR was studied . The essential role of metal ions in specific binding of alcohol dehydrogenase and dyes was demonstrated by differential spectroscopy, circular dichroism spectroscopy and chromatography . Lactate dehydrogenase in contrast with alcohol dehydrogenase does not require metal ions for the binding of the above-said dyes . A comparative study of eluting abilities of selected desorption agents (imidazole, adenine, 8-oxyquinoline-5-sulfonic acid, NAD, AMP, EDTA) by alcohol dehydrogenase chromatography on adsorbents with light-resistant yellow 2KT-Cu(II) and orange 5K revealed the differences in competition of the dyes for NAD-binding sites of alcohol dehydrogenase . The participation of light-resistant yellow 2KT-Cu(II) in the formation of mixed complexes with imidazole, adenine, 8-oxyquinoline-5-sulfonic acid, NAD and EDTA suggests that the specific binding of alcohol dehydrogenase to light-resistant yellow 2KT-Cu(II) is due to coordination between the Cu(II) ion and the amino acid residue in alcohol dehydrogenase.

FEBS Lett, 1987 Jan 1, 210(1), 66 - 70
Mitochondria catalyze the reduction of NAD by reduced methylviologen; Nagata S et al.; Mitochondria from beef heart and yeast catalyze the reduction of NAD to NADH at the expense of reduced methylviologen (MV+) . Based on protein the specific activity of mitochondria for this reaction is about 10 20-times higher than the consumption of oxygen in the presence of succinate or NADH . In 2H2O buffer (4S)-{4-2H}NADH is formed in high enantiomeric excess if the reduced methylviologen is electrochemically regenerated.

Drug Nutr Interact, 1987, 5(3), 169 - 79
Differential effects of dietary selenium on hepatic and renal glutathione-related enzymes and on hepatic microsomal drug metabolism in the rat; Davies MH et al.; These studies were undertaken to assess the effect of dietary selenium on glutathione-related enzyme activities in the liver and kidney and on hepatic drug metabolism . The intent was to study underlying mechanisms of selenium-induced beneficial effects in some models of hepatoxicity and carcinogenesis . Dietary selenium, as sodium selenite, was incorporated into a torula yeast basal diet (0.02 ppm selenium) and fed to male rats at supplementation levels of 0.0-5.0 ppm selenium for periods of three or six weeks . Additionally, a commercial cereal-based diet (CD, 0.05-0.08 ppm selenium) was compared to the experimentally defined diet (DD) supplemented with approximately the same amount of selenium . Liver and kidney glutathione peroxidase activity essentially plateaued at levels of selenium of 0.1 ppm and greater . CD- and DD-fed animals had hepatic and renal glutathione peroxidase activities which were similar . Glutathione S-transferase activity in liver, but not kidney, increased with increasing supplements of selenium . Glutathione S-transferase activities in CD- and DD-fed rats were not different . Cytochrome P-450 content and associated oxidative drug metabolism activities were relatively unmodified by selenium . Overall, dietary selenium appeared to act by enhancing potential conjugative detoxication pathway, rather than by decreasing the potential activation of chemicals via the hepatic cytochrome P-450 system.

Int Arch Allergy Appl Immunol, 1987, 84(4), 414 - 23
Mediators of C5a-induced bronchoconstriction in the guinea pig; Regal JF et al.; The effect of intravenous injection of C5a on pulmonary resistance and dynamic lung compliance was determined in anesthetized, artificially respirated guinea pigs . A mixture of C5a plus C5ades arg was purified from yeast-activated guinea pig serum and is referred to as C5a . Intravenous injection of C5a caused a dose-related bronchoconstriction as evidenced by a decrease in compliance and increase in resistance . Conversion of the C5a in the mixture to C5ades arg by carboxypeptidase B digestion did not significantly alter the magnitude of the bronchoconstriction . Pharmacological antagonists were employed to determine if histamine, acetylcholine or products of the arachidonate metabolism were mediators of C5a-induced bronchoconstriction . The histamine H1 antagonist pyrilamine inhibited the C5a-induced bronchoconstriction, suggesting the involvement of histamine . The cholinergic receptor antagonist atropine in combination with pyrilamine caused an inhibition of the C5a-induced increase in resistance but not compliance, suggesting acetylcholine does not play a major role in C5a-induced bronchoconstriction beyond its known role in participating in histamine-induced bronchoconstriction . Involvement of arachidonate metabolites was suggested by the ability of the cyclooxygenase inhibitor, indomethacin, to prevent the C5a-induced bronchoconstriction . Because indomethacin also caused a delay in the leukotriene C4 (LTC4)-induced bronchoconstriction, the participation of peptidoleukotrienes in the C5a-induced bronchoconstriction could not be ruled out . The leukotriene antagonists FPL 55712 and L-649,923 were evaluated for their specificity in inhibiting LTC4-induced bronchoconstriction . FPL 55712 was nonselective since it inhibited prostaglandin D2 and histamine-induced bronchoconstriction as well as LTC4-induced bronchoconstriction . L-649,923 inhibited only the LTC4-induced bronchoconstriction and was without effect on the C5a-induced bronchoconstriction, suggesting that peptidoleukotrienes are not important mediators of C5a-induced bronchoconstriction . Using radioimmunoassay, the change in peptidoleukotriene levels detected in plasma during C5a-induced bronchoconstriction was not significantly different from 0 . Thus, these studies have quantitated the C5a-induced decrease in dynamic lung compliance and increase in pulmonary resistance and suggest that histamine and cyclooxygenase products, but not peptidoleukotrienes, play a major role in C5a-induced bronchoconstriction.

Int Arch Allergy Appl Immunol, 1987, 84(4), 345 - 50
Hydrocortisone both decreases the up-regulation of complement receptors CR1 and CR3 and the ingestion process of human granulocytes; Forslid J et al.; We have studied the effect of hydrocortisone on the complement receptor expression (CR1 and CR3) of human granulocytes during the up-regulation phase and the following stable period when exposed to N-formyl-methionyl-leucyl-phenylalanine (fMLP) or the medium alone at 37 degrees C . The receptor expression was also correlated to the C3bi-mediated phagocytosis with special reference to attachment and ingestion . The results showed that after incubation with fMLP the increased expression of CR1 and CR3 is accompanied by an increased ingestion, but not attachment, of complement-opsonized yeast particles . This increase was significantly lower with regard both to attachment and ingestion, as well as receptor expression if hydrocortisone was present during the up-regulation phase . However, the addition of hydrocortisone after the up-regulation of fMLP-treated granulocytes decreased the ingestion of particles but not the CR1 and CR3 expression . The results indicate a membrane effect of hydrocortisone that affects both the receptor mobilization and ingestion mechanism of the phagocytes.

Z Erkr Atmungsorgane, 1987, 169(1), 23 - 32
{Chemiluminescent measurements of alveolar macrophages in sarcoidosis patients}; Winsel K et al.; The phagocytic activity of alveolar macrophages (AM) in a group of patients with stage I, stage II and stage III pulmonary sarcoidosis (gradation according to Wurm) has been investigated . Additionally a classification in different forms of the course of sarcoidosis was made (acute = Loefgren's syndrome, latent = primary-chronic, and relapses) . Patients with other lung diseases and healthy subjects were recruited as control group . The phagocytic activity (stimulation with opsonized yeast cell wall particles) of AM, which were isolated by bronchoalveolar lavage, was determined by means of chemiluminescence (CL)-measuring using lucigenin and luminol, respectively, as amplifiers . The investigations showed that the lucigenin-dependent yeast cell wall-induced CL of AM in patients with sarcoidosis is significantly increased in comparison to the control group . No significant changes of the luminol-dependent CL of AM from sarcoidosis patients could be detected . The lucigenin-dependent CL-response of AM is obviously an indicator of the intensity of the alveolitis and thus of the activity of the pathological process in sarcoidosis . The results suggest that in pulmonary sarcoidosis there is a hyperreactive AM/lymphocytes-system.

Dtsch Z Verdau Stoffwechselkr, 1987, 47(3), 128 - 33
{New knowledge of protein nitrogen resorption in the colon}; Heine W et al.; The absorption of protein nitrogen by the colon was assessed in 8 infants with colostomy by giving {15N} yeast protein in a dosage of 5-20 mg 15N/kg (92.4 atom-% 15N) . The absorption of 15N ranged between 87.1 and 98.1% of the administered dose, and the retention in the protein pool ranged between 81.0 and 95.7% . The incorporation of 15N in the plasma proteins was demonstrated by 15N-excess values between 0.01 and 0.10 atom-%, the TCA soluble fraction contained 15N-excess values of 0.04 to 0.19 atom-% . The results suggest that the colon can assimilate proteins when insufficient absorption of protein nitrogen in the small intestine occurs . The breakdown of protein is thought to result from the action of the colonic flora.

Nahrung, 1987, 31(5-6), 635 - 6
Lead and cadmium content in cocoa beans (short communication); Prugarova A et al.; The choice of cocoa beans as the experimental and sample material for study of the contamination with lead and cadmium was inspired by high Pb and Cd limits in foods made on its basis (cocoa powder, chocolate) as well as by the relatively high proportion of these foods in human nutrition . For Cd, the limits in food products are within the range of 0.01 mg X kg-1 (milk) to 1.0 mg X kg-1 (kidneys) whereas the limits for lead range between 0.1 mg X kg-1 (e.g . milk) and 10.0 mg X kg-1 (e.g . tea, yeast, crustaceans, molluscs) . Limits for Pb and Cd in foods made on cocoa bean basis are given in Table 1.

Chemotherapy, 1987, 33(4), 287 - 90
Effects of fluorouracil, doxorubicin and FAM combination on human peritoneal macrophages; Athlin L et al.; The effects of fluorouracil and adriamycin as single drugs and in combination with mitomycin (FAM) on the phagocytosis of tissue macrophages were studied in vitro . Human peritoneal macrophages (HPM) from cancer patients or uremic patients on peritoneal dialysis were incubated with the test drugs . No apparent inhibition of the adherence, engulfment or total phagocytosis of yeast cells was found with the fluorescence quenching technique . This is in contrast to our previous findings on blood monocytes . Thus the heterogeneity in sensitivity to anticancer drugs is probably related to cell maturity and cell surface characteristics.

Biomed Biochim Acta, 1987, 46(2-3), S209 - 13
Red blood cell glutathione peroxidase activity as a function of selenium supplementation in dietary treated children with phenylketonuria; Zachara BA et al.; Red blood cell glutathione peroxidase (GSH-Px) activity of six patients with phenylketonuria treated with aminoacid mixture and protein hydrolysate diets was significantly lower (11.2 IU/g Hb) than that of 21 age-matched healthy children (14.9 IU/g Hb) . When the diets were supplemented with yeast rich in selenium the red blood cell GSH-Px activity increased already after one month of treatment to 16.1 IU/g Hb (P less than 0.001) and remained at that level during subsequent two months of selenium supplementation . A high significantly positive correlation has been found between calculated red blood cell selenium level and GSH-Px activity within three months of selenium supplementation.

Exp Biol, 1987, 46(3), 141 - 7
Aerial respiration facilitates growth in suspension-feeding anuran larvae (Xenopus laevis); Wassersug RJ et al.; Xenopus laevis tadpoles are midwater suspension-feeders that use buccopharyngeal surfaces for both food capture and aquatic respiration, but also have functioning lungs well before metamorphosis . To examine the effect of aerial respiration on premetamorphic growth and development in this species, tadpoles were raised in normoxic water for from one to three weeks at three different concentrations of food (yeast cells in suspension) . At each food concentration the larvae were either allowed or denied access to air . Xenopus larvae reached greater snout-vent lengths, weighed more and developed more rapidly when allowed to breathe air . The difference in growth and development between those tadpoles with and without access to air was proportional to the concentration of particulate matter . In the extreme, tadpoles denied access to air at the highest food concentration all asphyxiated shortly after the start of the experiment . Xenopus tadpoles faced with the functional conflict of using buccopharyngeal surfaces for both feeding and respiration retard ingestion . They do this, however, not by ceasing mucus outflow to their branchial food traps, as has been speculated previously, but rather by capturing food particles in mucus and then expectorating it . Lungs appear to be advantageous to aquatic organisms even in normoxic water in that they allow buccopharyngeal surfaces to be dedicated fully to feeding rather than respiration.

Eur Biophys J, 1987, 14(5), 307 - 19
The superstructure of chromatin and its condensation mechanism . III: Effect of monovalent and divalent cations X-ray solution scattering and hydrodynamic studies; Koch MH et al.; Solutions of rat liver and chicken erythrocyte chromatin at different ionic strengths were characterized by synchrotron X-ray solution scattering, ultracentrifugation, density and viscosity measurements . Previous observations on nuclei were extended to rat liver, calf thymus and yeast nuclei . It is shown that with monovalent cations condensation is independent of the nature of the cation whereas with divalent cations there are significant differences related to the preference of base binding over phosphate binding . The consistency of hydrodynamic and scattering results confirm the view that chromatin in solution at low ionic strength has a helix-like superstructure . A survey of X-ray and neutron scattering results in the literature shows that previous interpretations, e.g . in terms of a 10 nm filament, are incompatible with the experimental data at low resolution.

J Cell Biochem, 1987 Jan, 33(1), 53 - 63
Evolution in the structure and function of aspartic proteases; Tang J et al.; Aspartic proteases (EC3.4.23) are a group of proteolytic enzymes of the pepsin family that share the same catalytic apparatus and usually function in acid solutions . This latter aspect limits the function of aspartic proteases to some specific locations in different organisms; thus the occurrence of aspartic proteases is less abundant than other groups of proteases, such as serine proteases . The best known sources of aspartic proteases are stomach (for pepsin, gastricsin, and chymosin), lysosomes (for cathepsins D and E), kidney (for renin), yeast granules, and fungi (for secreted proteases such as rhizopuspepsin, penicillopepsin, and endothiapepsin) . These aspartic proteases have been extensively studied for their structure and function relationships and have been the topics of several reviews or monographs (Tang: Acid Proteases, Structure, Function and Biology . New York: Plenum Press, 1977; Tang: J Mol Cell Biochem 26:93-109, 1979; Kostka: Aspartic Proteinases and Their Inhibitors . Berlin: Walter de Gruyter, 1985) . All mammalian aspartic proteases are synthesized as zymogens and are subsequently activated to active proteases . Although a zymogen for a fungal aspartic protease has not been found, the cDNA structure of rhizopuspepsin suggests the presence of a "pro" enzyme (Wong et al: Fed Proc 44:2725, 1985) . It is probable that other fungal aspartic proteases are also synthesized as zymogens . It is the aim of this article to summarize the major models of structure-function relationships of aspartic proteases and their zymogens with emphasis on more recent findings . Attempts will also be made to relate these models to other aspartic proteases.

EMBO J, 1987 Jan, 6(1), 35 - 41
A human and a plant intron-containing tRNATyr gene are both transcribed in a HeLa cell extract but spliced along different pathways; van Tol H et al.; tRNA splicing enzymes had been identified in mammalian and plant cells long before homologous intron-containing tRNA genes were detected . The tRNATyr gene presented here is the first intron-containing, human tRNA gene for which transcription and pre-tRNA maturation has been studied in a homologous system . This gene is disrupted by a 20-bp long intron and encodes one of the two major human tRNAsTyr which have been purified and sequenced . A tRNATyr gene recently isolated from Nicotiana also contains an intron and codes for a functional, major cytoplasmic tRNATyr . Both tRNA genes are efficiently transcribed in a HeLa cell nuclear extract . Each of them produces two independent primary transcripts because of two initiation and termination sites, respectively . The maturation of the tRNATyr precursors proceeds along different pathways . The intervening sequence of the human pre-tRNATyr is excised first, followed by ligation of the tRNA halves and maturation of the flanks, as has been shown for all intron-containing tRNA genes transcribed in HeLa extract . The order of maturation steps is reversed for the plant pre-tRNATyr: processing of the flanking sequences precedes intron excision . This maturation pathway corresponds to that observed in vivo for tRNA biosynthesis in Xenopus oocytes and yeast.

Drugs Exp Clin Res, 1987, 13(5), 237 - 45
A sulphonamido-indanone derivative CGP 28237 (ZK 34228), a novel non-steroidal anti-inflammatory agent without gastro-intestinal ulcerogenicity in rats; Bottcher I et al.; CGP 28237 (5-methylsulphonylamino-6-phenoxy-1-indanone) belongs to a series of structurally novel indanones . The compound is a weak acid (pK = 6.98), but it does not contain a carboxylic group . CGP 28237 exhibits potent anti-inflammatory activity in developing and established adjuvant arthritis in rats (ED40 approximately 0.5 mg/kg p.o.) and good activity in carrageenin oedema (ED40 approximately 3 mg/kg p.o.) . It inhibits yeast-induced fever in rats with ED50 values of 1, 2 and 10 mg/kg p.o . at 1, 3 and 5 hours after drug administration . The antinociceptive activity in mice (phenyl-p-benzoquinone writhing) and rats (acetic-acid writhing) is weak . CGP 28237 has been shown to be non-ulcerogenic in rats under acute and chronic test conditions: it does not cause mucosal lesions in the stomach at 2 X 400 mg/kg p.o., it does not enhance gastro-intestinal blood loss during 10 days' oral treatment with 400 mg/kg p.o., and it did not induce gastro-intestinal lesions in a 4-week toxicity study up to 1000 mg/kg p.o . Although CGP 28237 is not a cyclooxygenase inhibitor in bovine seminal vesicle microsomes, it inhibits prostaglandin synthesis in zymosan-stimulated murine macrophages (IC50 approximately 3 X 10(-6) mol/l) and protects rabbits against arachidonic acid-induced lung embolism with 10 mg/kg p.o . CGP 28237 may represent a novel anti-inflammatory drug with excellent gastro-intestinal tolerability.

J Reprod Fertil Suppl, 1987, 35, 599 - 605
Chemotactic and phagocytic function of peripheral blood polymorphonuclear leucocytes in newborn foals; Bernoco M et al.; Chemotactic and phagocytic responsiveness of peripheral blood polymorphonuclear leucocytes (PMNLs) from 11 foals were analysed immediately after birth (pre-colostral) and at different times after colostrum ingestion . The number of foal PMNLs per microscopic field that had migrated through the filter in chemotaxis and the number of yeast particles ingested per foal PMNL in phagocytosis were significantly lower when tested with foal plasma before colostrum ingestion (chemotaxis, 2.0 +/- 0.55 (s.e.m.); phagocytosis, 0.98 +/- 0.352) than in tests 4 or more days after colostrum ingestion (chemotaxis, 17.6 +/- 3.88; phagocytosis, 3.87 +/- 0.410; P less than or equal to 0.005) . A similar functional deficiency was found in foal PMNLs tested with pre-colostral foal plasma when compared to tests of the same cells with mare plasma (chemotaxis, 11.6 +/- 2.91; phagocytosis, 3.94 +/- 0.269; P less than or equal to 0.005) . PMNLs from neonatal foals responded with normal functions to plasma from normal adult horses . Results of both assays suggest that pre-colostral foal PMNLs are functionally mature but the expression of their chemotactic and phagocytic functions depends on the humoral source of chemotactic and opsonic factors.

Z Erkr Atmungsorgane, 1987, 169(3), 250 - 9
{Effect of antioxidants on the production of reactive oxygen metabolites by stimulated alveolar macrophages}; Winsel K et al.; Reactive oxygen metabolites (ROM) (O2-, H2O2, 1O2, .OH, OX-) which are produced by stimulated alveolar macrophages (AM), neutrophils and eosinophils, play an important role in the pathogenesis of many acute and chronic lung diseases . With regard to a therapeutic application the influence of the antioxidants ascorbic acid (Vitamin C) and alpha-tocopheryl acetate (Vitamin E acetate) on the production of ROM by AM was investigated . The AM were isolated by bronchoalveolar lavage from patients with different lung disorders . The ROM were determined by means of chemiluminescence-measuring . alpha-Tocopheryl acetate solved in peanut oil causes a little increase of the yeast cell wall-induced chemiluminescence . Pure alpha-Tocopheryl acetate has no effect on the chemiluminescence . In contrast to alpha-Tocopheryl acetate the addition Vitamin C to the stimulated AM results in a strong diminution of the chemiluminescence signal . This result suggests that Vitamin C reduces the generation of ROM by AM . Therefore Vitamin C could be a suitable scavenger of radicals and oxidants in different lung diseases.

Microbios, 1987, 52(212-213), 161 - 71
Media-induced departures from the usual, temperature-dependent cell shapes of Sporothrix schenckii and concomitant changes in the acid phosphatase isoenzyme patterns; Arnold WN et al.; Sporothrix schenckii grew as mycelia at 20 degrees C or yeast at 35 degrees C on a common culture medium, YNG (yeast extract, neopeptone and glucose), in agreement with previous observations that standard media support the mycelial phase at the lower temperature and the yeast phase at the higher temperature . Special media, M-1 and M-2, were used to generate yeast at 20 degrees C and mycelia at 35 degrees C, respectively, i.e . the reverse of the typically-observed, temperature-dependent phases . In this context the M-1 induced (or forced) generation of yeast cells at 20 degrees C was judged to be abnormal yet these cells had a typical yeast ultrastructure including the definitive microfibrillar zone of the cell envelope . Electropherograms of extracts of these cells (M-1 at 20 degrees C) displayed the typical acid phosphatase isoenzyme pattern of yeast grown at 35 degrees C, and that is much more complicated than the typical pattern for extracts of mycelia grown at 20 degrees C on YNG . On the other hand the fungus was held in the mycelial phase on M-2 at 35 degrees C, at least for short term cultures, to complement the study and provide a departure from the cell shape which is expected at the higher temperature . These mycelia were judged to be ultrastructurally abnormal in that they had a thin microfibrillar zone not previously seen in S . schenckii mycelia produced on standard media at lower temperatures . The cell-free extract of these abnormal mycelia grown at 35 degrees C exhibited an isoenzyme pattern which was closer to that of yeast grown at 35 degrees C than to mycelia grown at 20 degrees C, although one isoenzyme showed intermediate electrophoretic mobility . The findings with M-2 medium at 35 degrees C were less meaningful because of the eventual conversion to yeast phase . Overall the results indicate an association between acid phosphatase isoenzyme pattern and cell shape, rather than simply an expression of the various acid phosphatases with growth temperature.

Comp Biochem Physiol B, 1987, 88(2), 595 - 601
Properties of a ribonuclease from Aedes aegypti larvae; Fritz MA et al.; 1 . The properties of a soluble ribonuclease from Aedes aegypti larvae have been compared with ribonuclease activity in adult female tissue . 2 . In larval extracts ribonuclease activity was maximal at 40-45 degrees C whereas activity in tissue from adult females was highest at 50 degrees C . 3 . Ribonuclease activity that was recovered in a 20-60% ammonium sulfate precipitate was further purified by batch elution from DEAE-Sephacel and from carboxymethylcellulose . 4 . Ribonuclease activity in the partially purified fraction was sensitive to EDTA, stimulated by magnesium, had a pH optimum at 9.0 and a Mr of 45,000 . 5 . Agarose gels containing yeast RNA substrate were used to monitor partial purification of the larval ribonuclease.

Am J Chin Med, 1987, 15(3-4), 127 - 32
Evaluation of Artemisia inculta for anti-inflammatory activity in rats; Tariq M et al.; The ethanolic extract of Artemisia inculta has been screened for anti-inflammatory, analgesic and antipyretic activities on suitable experimental models . It has been found to produce significant inhibition of carrageenan induced paw edema and cotton pellet induced granuloma pouch and a significant decrease in the prothrombin time in rats . It failed to produce any analgesic or antipyretic activity on the hot plate reaction time and yeast induced hypyrexia tests in mice . It also did not produce any effect on the platelet aggregation and fibrinogen level in the rats . Amongst the phytoconstituents detected in this plant, flavonoids may be responsible for the observed anti-inflammatory effect of the ethanolic extract.

Symp Soc Exp Biol, 1987, 41, 113 - 33
Temperature and animal cell protein synthesis; Burdon RH; A predominant feature of brief heat stress to animal cells is the vigorous but transient activation of a small number of specific genes, previously either silent, or active at low levels . New mRNAs are actively transcribed from these genes and are translated into proteins, known collectively as the heat shock proteins, or hsps . The number of different types of hsp varies considerably in different organisms and cell types but in all cases proteins of approximately 84 and 70 kDa are amongst the most prominent . A dramatic feature that emerges from a study of their genes is that these proteins have been highly conserved during evolution . Gene sequence data reveal specific nucleotide sequences upstream of the transcription start sites that are essential for induction . These are known as 'heat shock elements' and are believed to be the region to which activated 'heat shock transcription factors' bind to facilitate hsp gene transcription . A recent model suggests that transcriptional regulation is based on competition between abnormal intracellular proteins and such a labile regulatory factor . Other experiments suggest such hsp inducers as heat, ethanol, arsenite, or oxygenation after anoxia, may cause protein damage through oxygen-derived free radical action . Although heat shock can cause very considerable changes in transcriptional patterns, effects specifically on translational control are no less dramatic . Different organisms achieve a rapid change in different ways . In Drosophila, heat shock promotes the translation of hsp mRNAs . In yeast there is no mechanism for sequestering pre-existing mRNAs from translation . Instead these mRNAs simply disappear rapidly from the cell . Mammalian cells are different in yet another way . There is neither sequestration of pre-existing mRNAs nor their removal from the cell . Superimposed upon the transient heat induced activation of hsp genes there are now clear indications of developmental regulation . The mechanism for the specific developmental control of hsp expression is not yet known . However, as oncogene products are known to induce hsp synthesis it may be that hsps are involved in eukaryotic growth control . Hypothermia causes loss of protein synthetic activity in cultured animal cells . No specific proteins are however induced and recovery at 37 degrees C is rapid.

Ann Biol Clin (Paris), 1987, 45(5), 553 - 7
{Recent advances in the biology of Candida}; Odds FC; There has been enormous progress in our understanding of biological processes in pathogenic Candida species, particularly in the case of C . albicans . Yeast taxonomists have continued to discover the sexual affinities of several members of the genus, although the sexual stage of C . albicans itself remains as elusive as ever . Genetically, C . albicans has been proved to be a diploid with balanced lethal mutations . It is likely to become a popular object of study for transposable genetic elements: recent research in the USA has demonstrated its ability to alter its phenotype at a very high frequency . Phenotypic switching is a property that may be associated with the relative virulence of the species . There is evidence for substantial phenotypic alteration of the cell surface of C . albicans in infected tissues in vivo, which may explain some of the pathological aspects of Candida infection . The best known of the "phenotypic switches" in C . albicans is its ability to transform from budding yeast cells into hyphae . Although the precise mechanisms of this change remains undiscovered, molecular and cell biological approaches to the problem are beginning to reveal many of the processes involved.

Crit Rev Microbiol, 1987, 15(1), 57 - 72
Anti-Candida drugs--the biochemical basis for their activity; Vanden Bossche H et al.; The past years have seen a continuous effort toward the synthesis of new antifungal agents . Most of them belong to the N-substituted imidazoles and triazoles . Another interesting series of antifungals are the allylamines . Biochemically, both the azole derivatives and the allylamines belong to the class of ergosterol biosynthesis inhibitors and thus differ from the polyene macrolide antibiotics . Indeed, it is now believed that the antifungal action of the polyenes, nystatin and amphotericin B, is due to a direct interaction with ergosterol itself . A more detailed analysis of the ergosterol biosynthesis inhibitors revealed that ergosterol depletion is the consequence of the interaction of the azole derivatives, e.g., miconazole, ketoconazole, and itraconazole, with the cytochrome P-450 involved in the 14 alpha-demethylation of lanosterol . Both the accumulation of 14 alpha-methylsterols and the concomitant decreased ergosterol content affect the membranes and membrane-bound enzymes of yeast and fungi . The allylamines seem to act by inhibition of the squalene epoxidase resulting in ergosterol depletion and accumulation of squalene . The target for the fluorinated pyrimidine, flucytosine, is completely different . Its antifungal properties may result from its conversion to 5-fluorouracil . The latter is then phosphorylated and incorporated into RNA, thus disrupting the protein synthesis in the yeast cell . These different biochemical targets for the antifungals of use in candidosis are discussed in this paper.

Gene, 1987, 53(2-3), 235 - 45
Nucleotide sequence of a cDNA for a member of the human 90-kDa heat-shock protein family; Rebbe NF et al.; This paper describes the isolation and sequence of a human cDNA homologous to a class of proteins commonly referred to as 90-kDa heat-shock proteins . The complete nucleotide sequence of 2563 bp and the deduced amino acid sequence are presented . A single long open reading frame encodes a protein of 83,303 Da, the amino acid composition of which correlates well with that determined for the human 90-kDa heat-shock or 'stress' protein {Welch, W.J . and Feramisco, J.R., J . Biol . Chem . 257 (1982) 14949-14959} . Moreover, sequence analysis of this gene reveals extensive homology with the Drosophila 83-kDa and yeast 90-kDa heat-shock proteins . A comparison of the translated product of the human cDNA to the published yeast 90-kDa heat-shock protein reveals more than 60% homology at both the nucleotide and amino acid levels . Several regions of 50 aa or more show greater than 90% identity . This cDNA also hybridizes with an RNA species which increases upon heat shock of HeLa cells.

Comp Biochem Physiol B, 1987, 87(1), 79 - 85
Purification and comparative studies of alcohol dehydrogenases; Tsai CS et al.; Alcohol dehydrogenases from various animal and plant sources were purified by a common procedure which employed DEAE, Sephadex-G100 and affinity chromatographies . The procedure achieves an 80-130 fold purification for animal enzymes . However, only a 5-15 fold purification for plant enzymes was attained because of the instability of these enzymes . Purified alcohol dehydrogenases from animal and plant sources differ in coenzyme and substrate specificities . The enzymes from mammalian, avian and fish livers display aldehyde oxidizing and esterolytic activities in addition to alcohol oxidizing activity . However, the enzymes from plants and yeast show only the oxidative activity toward alcohols . Chemical modifications have been performed to identify amino acid residues which are essential to the oxidative and esterolytic activities of alcohol dehydrogenases.

Gene, 1987, 51(2-3), 205 - 16
Comparison of the cis-acting control regions of two coordinately controlled genes involved in ethanol utilization in Aspergillus nidulans; Gwynne DI et al.; The alcA and aldA genes of Aspergillus nidulans are regulated in exactly the same manner, being subject to positive control by the product of the alcR gene . We report the complete nucleotide sequence of the alcA gene and its 5' non-coding region, preliminary localization of the region involved in the regulation of alcA expression, and a detailed comparison of this region to the 5' non-coding region of aldA (Pickett et al., 1987) . The 5' flanking regions of the genes contain six similar sequence elements . Three of these elements are located upstream from the messenger RNA start points and one is related to a sequence element found in the region responsible for ethanol induction of the yeast ADH2 gene (Beier et al., 1985) . The other homologous elements are located within the messenger RNA leader and may be associated with selection of messenger RNA start points . The amino acid sequence of alcohol dehydrogenase I (348 residues) shows a significant level of homology with analogous sequences in other organisms . Gene alcA contains introns which are similar in size and structure to other fungal introns . We discuss the positions of the introns in alcA of A . nidulans with particular reference to the conservation of intron position in and the evolutionary assembly of enzymes which possess NAD-binding domains.

Gene, 1987, 60(1), 115 - 27
Rapid construction of large synthetic genes: total chemical synthesis of two different versions of the bovine prochymosin gene; Wosnick MA et al.; We have tested several different synthesis designs and assembly methodologies to develop an improved gene synthesis strategy which enables significantly longer nucleotide sequences to be easily constructed . This strategy, based in part upon our ability to synthesize high-quality extended-length oligodeoxynucleotides (over 100-mer in length), together with the use of chemical 5'-phosphorylation, and simplified low-melting-temperature agarose gel purification methods, combines ease, speed and high overall efficiency . We show that it is now feasible to synthesize routinely even long genes (at least 1-2 kb) . To demonstrate this capability we have chemically synthesized and assembled two different versions of the gene encoding the bovine enzyme prochymosin (prorennin) . One gene is essentially the natural bovine prochymosin gene sequence . In the second gene the codons have been optimized with regard to the codon bias of highly expressed yeast genes . Each synthetic gene was in excess of 1100 bp, yet they were assembled from only 13 or 14 pairs of complementary oligodeoxynucleotides (oligos), the average lengths of which were 87 and 82 bp, respectively . The 'mutation' rate was low enough to assess that more than 75% of all such oligo pairs (160-170 total nt) were error-free.

Postgrad Med J, 1987, 63 Suppl 2, 27 - 32
Cost-benefit analysis of hepatitis B vaccination; Jonsson B; Cost effectiveness of hepatitis B vaccination is dependent on the vaccination strategy and the target group . Vaccination with plasma-derived vaccine has been shown to be cost-saving for high-risk groups such as homosexual men, and cost-effective for medium-risk populations such as surgical residents . For the population at large in European countries, vaccination is not a cost-effective use of scarce health care resources . Cost-benefit, cost-effectiveness, and cost-saving studies have made significant contributions to the design of rational vaccination strategies for hepatitis B vaccination programmes . These studies identify the cost of the vaccine and the infection rate to be the most important determinants of cost effectiveness . Studies of benefits and costs of vaccines generally undervalue the net benefits gained by prevention of pain and suffering associated with disease morbidity and mortality . As epidemiological conditions change, and with the introduction of the new generation of recombinant DNA yeast-derived hepatitis B vaccines, there is a need for repeated studies in different countries to identify the risk groups for which the expected net benefit of vaccination is positive . For such studies, improvements in both methodology and epidemiological data are needed.

J Mol Evol, 1987, 24(4), 337 - 45
Models of nearly neutral mutations with particular implications for nonrandom usage of synonymous codons; Li WH; The population dynamics of nearly neutral mutations are studied using a single-site and a multisite model . In the latter model, the nucleotides in a sequence are completely linked and the selection schemes employed are additive, multiplicative, and additive with a threshold . Although the third selection scheme is very different from the first two, the three schemes produce identical results for a wide range of parameter values . Thus the present study provides a general theory for the population dynamics of nearly neutral mutations because the results can also be used to draw inferences about other selection schemes such as stabilizing selection and synergistic selection . It is shown that the number of slightly deleterious mutations accumulated in a sequence can be considerably larger under the multisite model than under the single-site model, particularly if the sequence is long or if the mutation rate per site is high . The results show that even a very slight selective difference between synonymous codons can produce a strong bias in codon usage . Three alternative explanations for the strong bias in codon usage in bacterial and yeast genes are considered . The implications of the present results for molecular evolution are discussed.

Adv Dermatol, 1987, 2, 243 - 67
Wound healing for the clinician; Zitelli J; Wound healing is a complex sequence of events, beginning with tissue injury, mediated by inflammation, and ending long after reepithelialization is complete . Research and controlled clinical experience have provided a better understanding so that clinicians can influence the events of healing to decrease pain, control bleeding, infection, and cosmetic result as well as speed the time for complete healing . The following is a summary of guidelines for the management of wound healing: (1) wound creation; wounds should be created with minimal necrosis of tissue in order to prevent delays in healing . Electrosurgical, cryosurgical, and laser surgical wounds heal more slowly than wounds created by scalpel excision or curettage . Electro-coagulation should be used sparingly in sutured wounds . Large lesions are best treated in a single stage rather than in divided treatments since the rate of wound healing is not proportional to the area but instead to the logarithm of the area . Thus, the total healing time is much shorter if done in a single treatment session . (2) use of drugs; corticosteroids given before or within three days of wounding in dose of prednisone 40 mg or greater will inhibit wound healing . Vitamin A topically or systemically may reverse this inhibition . Aspirin and other nonsteroidal anti-inflammatory agents are more important for their effects on platelet function and bleeding than on wound healing . (3) wound dressings; the use of occlusive dressings to promote moist wound healing is the most significant advance in wound management . Occlusive dressings shorten the time for healing, decrease pain, reduce wound contamination, and improve the cosmetic result . (4) control of wound contraction and scar formation; at the time of wound formation, guiding sutures may be helpful in wound healing by secondary intention in order to control the direction of wound contraction and prevent distortion . Intralesional steroids may be useful for hypertrophic scars and keloids . (5) identification of complications; early identification of certain complications can prevent the delays in healing . These include infection, remembering infrequently cultured organisms such as yeast, malnourishment with protein and mineral deficiency, and the knowledge that adhesive-backed wound dressings can cause rewounding of otherwise normally healing wounds . (6) predicting the cosmetic result; wounds healed by secondary intention may provide a cosmetic result superior to surgical repair . Wounds in concave areas usually heal with a better result than wounds managed by flaps or grafts although wounds on convex surfaces usually look best if a skillful primary closure can be performed without distortion.(ABSTRACT TRUNCATED AT 400 WORDS)

J Mol Evol, 1987, 25(1), 65 - 73
The sequence of the 3.3-kilobase repetitive element from Dipodomys ordii suggests a mechanism for its amplification and interspersion; Keim P et al.; DNA from the kangaroo rat, Dipodomys ordii, contains a 3.3-kb, highly repeated sequence that is interspersed throughout the genome in small tandem clusters . One 3.3-kb unit has been cloned into pBR322 and the nucleotide sequence determined . The clone used was shown to be representative of the bulk of such sequences found in the genomic DNA . The sequence contains 10 homologous subunits each ca . 260 bp in length . Comparison of these to one another yielded a 258-bp consensus sequence containing a 35-bp terminal inverted repeat . Two unique stretches also occur . One of these contains a region that could serve as a promoter for RNA polymerase III; the other contains a sequence related to the ARS sequences of yeast . It is proposed that an ancestral sequence similar to the consensus sequence was amplified to 10 or more units, and that, subsequently, two other sequences were inserted . The properties of these insertions may have led to the dispersal of the sequence throughout the genome.

Comp Biochem Physiol B, 1987, 86(3), 547 - 54
Metabolism of glycerate 2,3-P2--XI . Essential amino acids of pig phosphoglycerate mutase isozymes; Berrocal F et al.; Phosphoglycerate mutase isozymes (types M, B and MB) from pig tissues are inactivated upon treatment with reagents specific for histidyl, arginyl and lysyl residues . Their mutase, 2,3-bisphosphoglycerate synthase and 2,3-bisphosphoglycerate phosphatase activities are concurrently lost, although some differences exist in the rate of inactivation . No significant differences are observed between the isozymes . The reversion of the modifying reactions reactivates the three enzymatic activities . Substrates and cofactors protect against inactivation, the protective effects varying with the modifying reagent . Titration with pCMB shows the existence of two essential thiol groups per subunit type M . These results provide evidence of the intrinsic character of the three enzymatic activities, favor their location at the same active site and suggest the existence of separate binding sites for monophosphoglycerates and bisphophoglycerates . Both type M and B subunit from pig phosphoglycerate mutase are similar to type M subunit from rabbit and to the enzyme from yeast.

Xenobiotica, 1987 Jan, 17(1), 45 - 57
Interaction of miconazole, ketoconazole and itraconazole with rat-liver microsomes; Lavrijsen K et al.; The interaction of the antimycotics miconazole, ketoconazole and itraconazole with liver microsomes from untreated rats or from rats pretreated with phenobarbital or 3-methylcholanthrene, gave rise to type II difference spectra . The interactions of the antimycotics with control, phenobarbital-induced or 3-methylcholanthrene-induced microsomes were biphasic, except for the monophasic binding of ketoconazole to phenobarbital-induced microsomes . The N-demethylation of N,N-dimethylaniline, the O-demethylation of p-nitroanisole and the hydroxylation of aniline in microsomes from untreated and inducer-treated rats were lowered by miconazole and ketoconazole, the former being the more potent inhibitor . Control microsomes were less sensitive than induced microsomes . Itraconazole was almost devoid of inhibitory properties . The three antimycotics were non-competitive (mixed) inhibitors of enzyme activities in phenobarbital-induced microsomes . The Ki values were of the same order of magnitude as the Ks values, except for itraconazole . For the latter drug, Ki values were much greater than could be expected from the spectral studies . It is concluded that the antimycotics affect microsomal enzyme activities via a direct interaction of an azole-nitrogen with the haem group of cytochrome P-450 . The interaction with mammalian cytochrome P-450 decreases from miconazole greater than ketoconazole much greater than itraconazole and is much weaker than the interaction of the antimycotics with yeast cytochrome P-450.

J Neurosci, 1987 Jan, 7(1), 77 - 87
A model of chronic pain in the rat: functional correlates of alterations in the activity of opioid systems; Millan MJ et al.; Intradermal inoculation of rats at the tail base with Mycobacterium butyricum led to the gradual development of an arthritic swelling of the limbs which peaked at 3 weeks and subsided thereafter . Arthritic rats displayed a loss of body weight, hypophagia, and hypodipsia in addition to a disruption of the diurnal rhythms of ingestive behavior and of core temperature . The activity of adenohypophyseal beta-endorphin-(beta-EP) secreting corticotrophs, in contrast to prolactin-(PRL) secreting lactotrophs, was increased in arthritic rats . Indeed, hypertrophy of the adrenal glands was seen . Arthritic rats also showed an elevation in spinal cord levels of immunoreactive dynorphin (DYN), an endogenous ligand of the kappa-opioid receptor . The paws and tail of arthritic rats showed lower thresholds in response to noxious pressure (hyperalgesia), higher thresholds in response to noxious heat (hypoalgesia), and no change in their response to noxious electrical stimulation . Neither naloxone nor ICI-154, 129 (a preferential delta-receptor antagonist) modified the responses of the paw or tail to pressure . However, MR 2266 (an antagonist with higher activity at kappa-receptors) decreased thresholds to pressure in arthritic, but not control, rats; that is, it potentiated the hyperalgesia . This action was stereospecific . None of the antagonists modified the response to heat . MR 2266 did not affect the response to pressure in rats with acute inflammation produced by yeast . Thus, the potentiation of pressure hyperalgesia by MR 2266 in chronic arthritic rats is highly selective . Arthritic rats showed a reduced response to the analgesic effect of a kappa-agonist (U-50,488H), whereas the response to a mu-agonist (morphine) was enhanced . These effects were specific to nociception in that their influence upon endocrine secretion (PRL and beta-EP) was otherwise changed . The secretion of beta-EP and PRL was stimulated by both morphine and U-50,488H, and the influence of U-50,488H upon the release of beta-EP (from the adenohypophysis) was enhanced in arthritic rats . It is suggested that polyarthritis is a complex condition entailing many changes, both behavioral and endocrinological . Further, arthritic rats cannot simply be described as "hyperalgesic": of critical importance is the nature of the nociceptive stimulus applied . The parallel alterations in spinal cord pools of DYN and kappa-receptors (see also Millan et al., 1986) and the changes in the influence on nociception of kappa-agonists and kappa-antagonists suggest an increased activity of spinal DYN . Thus, spinal kappa-receptors may play a role in the modulation of nociception under chronic pain.(ABSTRACT TRUNCATED AT 400 WORDS)

Antiviral Res, 1987 Jan, 7(1), 43 - 51
Photodynamic inactivation of influenza and herpes viruses by hematoporphyrin; Perlin M et al.; Hematoporphyrin (HP), at concentrations as low as 0.5 microgram/ml, was found to inhibit the in vitro replication of influenza A and herpes simplex viruses, but not of several other viruses . The effect required exposure of the viruses or cells to visible light and was demonstrable when HP was administered shortly before virus inoculation or during the infection . In studies on the mechanism of action of HP, we found that in the presence of light, HP caused decomposition of GMP but not of various other nucleosides . It caused breakdown of yeast tRNA and inhibited polymerization of RNA and DNA by influenza virus and HSV-1-specific polymerases as well as some other polymerases isolated from bacterial and mammalian sources . Protective effects of HP and light were demonstrable in embryonated eggs infected with the WSN and PR8 strains of influenza A virus and in mice infected with the WSN strain . HSV-1-induced keratitis in rabbits and HSV-2-induced dermatitis in mice were not responsive to HP treatment.

Proc Natl Acad Sci U S A, 1987 Jan, 84(1), 194 - 8
Nucleotide sequence of a preferred maize chloroplast genome template for in vitro DNA synthesis; Gold B et al.; Maize chloroplast DNA sequences representing 94% of the chromosome have been surveyed for their activity as autonomously replicating sequences in yeast and as templates for DNA synthesis in vitro by a partially purified chloroplast DNA polymerase . A maize chloroplast DNA region extending over about 9 kilobase pairs is especially active as a template for the DNA synthesis reaction . Fragments from within this region are much more active than DNA from elsewhere in the chromosome and 50- to 100-fold more active than DNA of the cloning vector pBR322 . The smallest of the strongly active subfragments that we have studied, the 1368-base-pair EcoRI fragment x, has been sequenced and found to contain the coding region of chloroplast ribosomal protein L16 . EcoRI fragment x shows sequence homology with a portion of the Chlamydomonas reinhardtii chloroplast chromosome that forms a displacement loop {Wang, X.-M., Chang, C.H., Waddell, J . & Wu, M . (1984) Nucleic Acids Res . 12, 3857-3872} . Maize chloroplast DNA fragments that permit autonomous replication of DNA in yeast are not active as templates for DNA synthesis in the in vitro assay . The template active region we have identified may represent one of the origins of replication of maize chloroplast DNA.

J Exp Med, 1987 Jan 1, 165(1), 195 - 210
Role of the adherence-promoting receptors, CR3, LFA-1, and p150,95, in binding of Histoplasma capsulatum by human macrophages; Bullock WE et al.; The principal host cell of H . capsulatum (Hc) is the M phi within which the pathogenic yeast phase of the fungus multiplies during active disease . The initial interaction between Hc yeasts and M phi therefore is a crucial step in the pathogenesis of histoplasmosis . In the present study, we have identified the major receptor mechanism that mediates the attachment of unopsonized Hc yeasts to human monocyte-derived M phi from peripheral blood . Binding of Hc yeasts by M phi is rapid, temperature dependent, and requires both Ca and Mg ions for optimum activity . Recognition of Hc yeasts does not require Fc receptors, mannosyl/fucosyl receptors, beta-glucan receptors, or secretion of C3 by M phi . Studies were performed on the effect of down regulating specific receptors of the CR3/LFA-1/p150,95 adherence-promoting protein family from the apical portion of M phi to determine the effects upon binding of Hc yeasts . Anti-beta chain mAbs that recognize all three of these proteins blocked binding of yeasts . However, removal of individual receptors with antibodies against the alpha polypeptides caused negligible depression of binding, and removal of any pair caused only modest depression . Thus, each of the members of the CR3/LFA-1/p150,95 family is independently capable of binding Hc . The delineation of this new mechanism for nonopsonic recognition by M phi that is exploited by Hc yeasts will aid in future studies to identify the Hc ligand, to elucidate the stoichiometry of CR3/LFA-1/p150,95 binding, and to determine triggering mechanisms for release of toxic oxygen metabolites.

Infect Immun, 1987 Jan, 55(1), 29 - 34
Histoplasma capsulatum fails to trigger release of superoxide from macrophages; Eissenberg LG et al.; The yeast form of the dimorphic fungus Histoplasma capsulatum survives within macrophages after phagocytosis . To do so, it must avoid, inhibit, or resist a variety of toxic oxygen metabolites . Using ferricytochrome c reduction to assay superoxide release, we examined the response of mouse macrophages to the yeast form of various H . capsulatum strains . Doses of zymosan as low as 20 particles per macrophage elicited superoxide, whereas H . capsulatum failed to induce superoxide even at 160 yeast cells per macrophage . This phenomenon was observed with two virulent strains of H . capsulatum (G217B and G186A) and with an avirulent variant of G186A . Over a 15- to 150-min observation period, zymosan stimulated increasing reduction of ferricytochrome c, but H . capsulatum did not . When added concurrently with zymosan, H . capsulatum had no effect on superoxide production . Therefore, H . capsulatum was unable either to inactivate the oxygen radical or inhibit host cell superoxide response to other competent stimuli . Enzymatically generated superoxide reduced ferricytochrome c even in the presence of H . capsulatum, again implying that the organism does not readily inactivate superoxide . This experiment also demonstrated that the yeast did not interfere with the assay used . Thus, rather than inhibiting superoxide generation or inactivating the anion, H . capsulatum yeast cells appear to avoid the toxic effects of superoxide by failing to trigger its release.

J Membr Biol, 1987, 98(2), 169 - 89
Potassium-proton symport in Neurospora: kinetic control by pH and membrane potential; Blatt MR et al.; Active transport of potassium in K+-starved Neurospora was previously shown to resemble closely potassium uptake in yeast, Chlorella, and higher plants, for which K+ pumps or K+/H+-ATPases had been proposed . For Neurospora, however, potassium-proton cotransport was demonstrated to operate, with a coupling ratio of 1 H+ to 1 K+ taken inward so that K+, but not H+, moves against its electrochemical gradient (Rodriguez-Navarro et al., J . Gen . Physiol . 87:649-674) . In the present experiments, the current-voltage (I-V) characteristic of K+-H+ cotransport in spherical cells of Neurospora has been studied with a voltage-clamp technique, using difference-current methods to dissect it from other ion-transport processes in the Neurospora plasma membrane . Addition of 5-200 microM K+ to the bathing medium causes 10-150 mV depolarization of the unclamped membrane, and yields a sigmoid I-V curve with a steep slope (maximal conductance of 10-30 microS/cm2) for voltages of -300 to -100 mV, i.e., in the normal physiologic range . Outside that range the apparent I-V curve of the K+-H+ symport saturates for both hyperpolarization and depolarization . It fails to cross the voltage axis at its predicted reversal potential, however, an effect which can be attributed to failure of the I-V difference method under reversing conditions . In the absence of voltage clamping, inhibitors-such as cyanide or vanadate-which block the primary proton pump in Neurospora also promptly inhibit K+ transport and K+-H+ currents . But when voltage clamping is used to offset the depolarizing effects of pump blockade, the inhibitors have no immediate effect on K+-H+ currents . Thus, the inhibition of K+ transport usually observed with these agents reflects the kinetic effect of membrane depolarization rather than any direct chemical action or the cotransport system itself . Detailed study of the effects of {K+}o and pHo on the I-V curve for K+-H+ symport has revealed that increasing membrane potential systematically decreases the apparent affinity of the transporter for K+, but increases affinity for protons (Km range: for {K+}o, 15-45 microM; for {H+}o, 10-35 nM) . This behavior is consistent with two distinct reaction-kinetic models, in which (i) a neutral carrier binds K+ first and H+ last in the forward direction of transport, or (ii) a negatively charged carrier (-2) binds H+ first and K+ last.

Antonie Van Leeuwenhoek, 1987, 53(3), 171 - 81
Stabilization of chitin synthetase and purification of chitosomes from several mycelial Mucorales; Martinez-Cadena G et al.; Stability of chitin synthetase in cell-free extracts from mycelial fungi was markedly improved by the presence of sucrose in the homogenization media . Breakage of mycelium in sucrose-containing buffer yielded enzyme preparations from which chitosomal chitin synthetase could be purified by a procedure involving ammonium sulfate precipitation, gel filtration and centrifugation in sucrose density gradients . Purified chitosomes catalyzed the synthesis of chitin microfibrils in vitro upon incubation with substrate and activators . Chitosomal chitin synthetase from the filamentous form of M . rouxii was similar to the enzyme from yeast cells, except for the poorer stability and diminished sensitivity to GlcNAc activation of the former.

Ann Biol Clin (Paris), 1987, 45(6), 661 - 8
{Anti-Candida activity of azoles}; Van Cutsem J et al.; The in vitro activity of the broad-spectrum antifungals miconazole, ketoconazole and itraconazole was evaluated by the decimal dilution method in liquid media, for respectively 2511, 1536 and 1859 strains of yeasts belonging to 31 species . Sabouraud broth was used for miconazole and brain heart infusion broth for ketoconazole and itraconazole . These three azoles proved to be potent anti-yeast compounds . Activity by oral treatment of miconazole, ketoconazole and itraconazole was compared in gastro-intestinal candidosis of the guinea-pig, in vaginal candidosis of the rat and in systemic candidosis of the guinea-pig . Ketoconazole and itraconazole are more efficacious than miconazole in gastro-intestinal candidosis . Miconazole showed marginal activity in the other two experimental models: the spectrum of activity of miconazole was a lead for research which resulted in the synthesis and selection of ketoconazole and itraconazole . Ketoconazole was highly active in these experimental models . Itraconazole cured the animals at distinctly lower doses . No side-effects due to these azoles were observed during these experiments . A proposal is made to classify the broad-spectrum azoles in three classes: compounds for topical use (e.g . miconazole), compounds with oral activity without activity in aspergillosis (e.g . ketoconazole), compounds with oral activity including activity in aspergillosis (e.g . itraconazole).

Acta Derm Venereol, 1987, 67(5), 445 - 7
Experimental folliculitis with Pityrosporum orbiculare: the influence of host response; Goodfield MJ et al.; The aetiology of the folliculitis associated with seborrhoeic eczema is unclear, though the yeast, Pityrosporum orbiculare has been implicated . P . orbiculare was applied under occlusion to normal forearm skin of patients with seborrhoeic eczema (SE), seborrhoeic eczema and folliculitis (SEF), and normal controls . There were significant differences in response to occlusion between the three groups . Those patients with previous clinical evidence of folliculitis (SEF) developed folliculitis at the site of occlusion more frequently than either of the other two groups (p less than 0.001), in whom only one patient developed skin changes . This difference was not explained by the response to occlusion alone, nor by natural carriage of yeasts . These results suggest that the yeast P . orbiculare is necessary for the development of folliculitis, but that the nature of the host response determines those patients prone to follicular inflammation.

Gene, 1987, 56(1), 29 - 40
Cloning and nucleotide sequence of the murine hsp84 cDNA and chromosome assignment of related sequences; Moore SK et al.; The nucleotide (nt) sequence of mouse 84-kDa heat shock protein (Hsp) cDNA has been determined using a combination of molecular cloning and oligodeoxynucleotide priming on poly(A) + RNA . The cDNA was 2.5 kb long, not including the poly(A) tail . It contained a 5' leader of about 94 nt that was G + C-rich, and a 243-nt 3'-untranslated region that was A + T-rich in the vicinity of the polyadenylation signal . Gene hsp84 codes for an acidic polypeptide of 724 amino acid (aa) residues . Mouse Hsp84 had 81% and 63% aa homology to Drosophila melanogaster Hsp82 and yeast Hsp90, respectively . The nucleotide sequence had 74% and 59% homology to Drosophila and yeast hsp sequences, respectively, in the coding regions of these genes . This homology did not extend to the 5' - and 3'-untranslated regions . Chromosomal analysis indicated that hsp84-related sequences are on at least three different chromosomes.

Cell, 1986 Dec 26, 47(6), 965 - 71
Binding and cleavage of pre-tRNA by the Xenopus splicing endonuclease: two separable steps of the intron excision reaction; Baldi MI et al.; We have constructed three base-substitution mutants of the yeast tRNALeu3 gene . In two of them the ability to form an extended anticodon stem is lost . In the first mutant the bases encoding the anticodon change from TTG to GAC (positions 37, 36, 35); in the second, the nucleotides encoding the region of the intron that base-pair with the anticodon change from CAA to GTC (positions 48, 47, 46) . The third is a double mutant characterized by both substitutions described above so that its ability to form an extended anticodon stem is restored . The precursors derived from the two single mutants are accurately spliced in the X . laevis germinal vesicles (GV) extract: pairing of the anticodon with the intron, therefore, is not required for the splicing reaction . The precursor derived from the double mutant is not spliced, indicating that the new extended anticodon stem exerts an inhibitory action . Since the double mutant precursor binds to the purified splicing endonuclease, binding and cleavage occur as two separable steps in the intron excision reaction.

Nucleic Acids Res, 1986 Dec 22, 14(24), 9549 - 59
A catalogue of splice junction and putative branch point sequences from plant introns; Brown JW; Splice junction and possible branch point sequences have been collected from 177 plant introns . Consensus sequences for the 5' and 3' splice junctions and for possible branch points have been derived . The splice junction consensus sequences were virtually identical to those of animal introns except that the polypyrimidine stretch at the 3' splice junction was less pronounced in the plant introns . A search for possible branch points with sequences related to the yeast, vertebrate and fungal consensus sequences revealed a similar sequence in plant introns.

Eur J Biochem, 1986 Dec 15, 161(3), 695 - 700
Intracellular pathway followed by invertase endocytosed by rat liver; Jadot M et al.; Yeast invertase, when injected into rats, is endocytosed by the liver, mainly by sinusoidal cells . The work reported here aims at investigating the organelles involved in the intracellular journey of this protein . Experiments were performed on rats injected with 125I-invertase (25 micrograms/100 g body wt) and killed at various times after injection . Homogenates were fractioned by differential centrifugation, according to de Duve, Pressman, Gianetto, Wattiaux and Appelmans {(1955) Biochem . J . 63, 604-617} . Early after injection the radioactivity was recovered mainly in the microsomal fraction P; later it was found in the mitochondrial fractions (ML) . At all times a peak of relative specific activity was observed in the light mitochondrial fraction L . After isopycnic centrifugation in a sucrose gradient, structures bearing 125I-invertase, present in P, exhibited a relatively flattened distribution with a density of around 1.17 g/ml, relatively similar to that of alkaline phosphodiesterase a plasma membrane marker . The organelles located in ML were endowed with a more homogeneous distribution, their median equilibrium density increasing up to 30 min after injection (1.20 g/ml----1.23 g/ml); with time the radioactivity distribution became more closely related to the distribution of arylsulfatase, a lysosomal enzyme . ML fractions, isolated 10 min and 180 min after 125I-invertase injection, were subjected to isopycnic centrifugation in Percoll gradient with, as solvent, 0.25 M, 0.5 M and 0.75 M sucrose . The change of density of the particles bearing 125I-invertase, as a function of the sucrose concentration, paralleled the change of density of the lysosomes as ascertained by the behaviour of arylsulfatase . The distribution of radioactivity and arylsulfatase in a sucrose gradient was established after isopycnic centrifugation of the ML fraction of rats injected with 125I-invertase, the animals having received or not an injection of 900 micrograms/100 g body weight of unlabelled invertase 15 h before killing . In agreement with our previous results, a shift towards higher densities of about 25% or arylsulfatase takes place in rats pretreated with unlabelled invertase . At 10 min, invertase preinjection did not change the radioactivity distribution curve . Later, it caused a progressive shift of the distribution towards higher-density regions of the gradient where the arylsulfatase, which had been shifted, was located.(ABSTRACT TRUNCATED AT 400 WORDS)

Nature, 1986 Dec 11-17, 324(6097), 589 - 91
Effect of mutations at the lariat branch acceptor site on beta-globin pre-mRNA splicing in vitro; Hornig H et al.; Introns are excised from full-length transcripts (pre-messenger RNAs) of eukaryotic genes in two steps . First, the pre-mRNA is cleaved at the 5' splice site and a branched (lariat) intermediate is formed . Then, cleavage at the 3' splice site and ligation of the two exons leads to the release of the lariat intron . The intron sequence which accepts the 5' end to form the lariat branch is strictly conserved in yeast, but shows more variation in eukaryotes . To investigate the requirements for branch formation in eukaryotes further, we have studied in vitro splicing of a rabbit globin gene intron with mutations of the normal branch-accepting adenosine nucleotide . We conclude that all four nucleotides can serve as branch acceptors, but that A and C are preferred to G and U in lariat formation . Mutation of the normal A to G or U can lead to an A residue one nucleotide upstream of the normal branch site being used instead . Only branches to A or C participate efficiently in the second splicing step.

J Biol Chem, 1986 Dec 5, 261(34), 16215 - 8
Crystallographic characterization of recombinant human CuZn superoxide dismutase; Parge HE et al.; Recombinant human CuZn superoxide dismutase as expressed in yeast has been crystallized in three different crystal forms . Hexagonal plates grow from 2.4 M ammonium sulfate, pH 7.5, and belong to the space group P6(3)22, with cell dimensions a = b = 113.5(3), c = 151.5(5) A, and Vm = 2.21 A3/dalton for two dimers per asymmetric unit . At 2.0 M ammonium sulfate, pH 7.5, chunky wedges grow in space group C222(1), a = 205.2(6), b = 166.5(4), c = 145.4(4) A with a Vm of 2.43 A3/dalton for eight dimers per asymmetric unit . With polyethylene glycol 8000, pH 7.5-8.0, hexagonal prisms are obtained with cell dimensions a = b = 197.4(6), c = 43.1(2) A, space group P6, and Vm = 2.53 A3/dalton for three dimers per asymmetric unit . All of these forms diffract to high resolution, are stable to x-rays, and appear suitable for determination of the atomic structure . Crystals of the doubly mutated enzyme (Cys6----Ala, Cys111----Ser) grown from both micro- and macroseeds of the wild type protein demonstrate the feasibility of isomorphous crystallization of site-directed mutants of the cloned parent enzyme for comparative structure-function studies.

Br J Pharmacol, 1986 Dec, 89(4), 719 - 30
Kinetics of the generation and action of chemical mediators in zymosan-induced inflammation of the rabbit peritoneal cavity; Forrest MJ et al.; Acute inflammation was induced by intraperitoneal injection of zymosan (yeast cell walls) in the rabbit . Peritoneal inflammation was monitored by the local accumulation of intravenously-injected Evans blue dye (which binds to plasma albumin) and of polymorphonuclear leukocytes (PMNLs) . The zymosan-induced exudate fluid contained a microvascular permeability-increasing factor or factors which, unlike histamine and bradykinin, had a long duration of action when tested in rabbit skin and was dependent on circulating PMNLs . Using radioimmunoassay, high levels of rabbit C5a, or C5a des Arg, were detected in the exudate fluid and accounted for much of the permeability-increasing activity, as judged by skin bioassay after separation on Sephadex G-100 . The vasodilator prostaglandin, prostaglandin I2 (PGI2), was generated in the inflammatory reaction, as judged by the presence of high levels of 6-oxo-PGF1 alpha detected in the exudate by radioimmunoassay . However, in contrast to observations in rabbit skin, inhibition of prostaglandin generation had a relatively small effect on peritoneal oedema formation . C5a and C5a des Arg increase microvascular permeability by a PMNL-dependent mechanism in the rabbit . However, in response to zymosan, protein leakage was detected considerably earlier than PMNL accumulation . A hypothesis to account for this difference is proposed.

Mycopathologia, 1986 Dec, 96(3), 143 - 51
Increased resistance of splenectomized mice to Sporothrix schenckii infection; Miyaji M et al.; The susceptibility of splenectomized mice to Sporothrix schenckii was studied, and the role of the spleen in the host defense is discussed . S . schenckii Sp-1 and ddy male mice were used . The mice were divided into 3 groups consisting of splenectomized, sham-operated and intact mice . Each mouse was inoculated intravenously with 2 X 10(6) yeast cells 7 days after operation and the mice were sacrified at adequate intervals for 30 days . Then histological sections stained with H&E or by PAS were prepared from various visceral organs . Using the liver sections the number of yeast cells in a 40 mm2 was counted . Furthermore, the colony forming unit in 100 mg of the liver tissue was compared to each other . In the sham-operated and intact mice many purulent lesions appeared on the 5th day . On the 8th day mononuclear cells accumulated at the foci, and on the 10th day most of the foci became granulomatous . The number of yeast cells in granulomatous lesions reached a peak on the 10th day and thereafter decreased abruptly . On the other hand, in the splenectomized mice approximately half of foci became granulomatous on the 5th day, and the number of yeast cells in the foci began to decrease after the 5th day . There were definite differences in the colony forming unit between the splenectomized and sham-operated or intact mice sacrificed 9 days after inoculation . The colony forming unit of the former is 9.3 X 10(5) on the average, while that of the latter two is 5.6 X 10(6) and 5.1 X 10(6) on the average, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

J Clin Invest, 1986 Dec, 78(6), 1638 - 47
Correlation between pathogenicity and temperature sensitivity in different strains of Histoplasma capsulatum; Medoff G et al.; We compared the mycelial to yeast transitions of the Downs strain of Histoplasma capsulatum (low level of virulence) with those of G184A and G222B, two more virulent strains having different levels of pathogenicity for mice . When the morphological transitions are initiated by a temperature shift from 25 degrees to 37 degrees C, all three strains undergo similar physiological changes, but these are less severe in G184A and G222B than in the Downs strain . The transitions from mycelial to yeast morphology in both of the more virulent strains are also one-third more rapid than in Downs . We also find that the differences in temperature sensitivity of the three strains can be correlated with the temperature required for complete uncoupling of oxidative phosphorylation . The differences in sensitivity to elevated temperatures extend to the growth of yeast cells of all three strains . Considered together, our results suggest that sensitivity to elevated temperatures may be a key factor accounting for differences in virulence and that uncoupling of oxidative phosphorylation may be the primary event in the morphological transition in all three strains.

Environ Res, 1986 Dec, 41(2), 488 - 96
Rabbit alveolar macrophages after long-term inhalation of soluble cobalt; Johansson A et al.; Rabbits were exposed to 2 or 0.4 mg/m3 of cobalt as CoCl2 for 14-16 weeks (5 days/week and 6 hr/day) . More macrophages were lavaged from the lungs of rabbits exposed to the higher Co2+ concentration, and the diameter and variation of the diameter of the macrophages were significantly larger than in controls . The activity of lysozyme in the lavage fluid and in the macrophages was increased in the two exposed groups . Some macrophages in the exposed animals were large and engorged with intracellular lamellar inclusions and lipid droplets . Most of these cells had a smooth surface . The oxidative metabolic activity measured by reduction of nitroblue tetrazolium was increased in the exposed groups . The number of yeast cell particles attached to the surface of the macrophages was increased in the group exposed to the high concentration, but the number of ingested particles was not affected by cobalt exposure . Apart from the fact cobalt increased lysozyme activity whereas nickel decreased it, cobalt produced the same type of effects on macrophages as nickel did in earlier studies . Cobalt affected only a minor proportion whereas nickel affected most macrophages . This can be explained by the fact nickel produced a general increase in the volume density of the type II cells while cobalt affected the type II cells only in some areas of the lungs.

Arch Pathol Lab Med, 1986 Dec, 110(12), 1180 - 1
Candida parapsilosis diagnosed by peripheral blood smear; Monihan JM et al.; A 52-year-old man presented with a history of chronic lymphocytic leukemia and Kaposi's sarcoma . Three days after admission for clinical sepsis, yeast forms were noted within monocytes and neutrophils on a routine peripheral blood smear . Blood cultures yielded a pure culture of Candida parapsilosis . Despite antifungal therapy, the patient died 22 days after admission . This case represents the first reported instance of C parapsilosis sepsis diagnosed through a peripheral blood smear.

J Gen Microbiol, 1986 Dec, 132 ( Pt 12), 3421 - 32
Acid phosphatases of Sporothrix schenckii; Arnold WN et al.; Sporothrix schenckii cells were grown on a medium containing yeast extract, neopeptone and glucose at 20 degrees C to obtain a mixture of mycelia and conidia, and at 35 degrees C to obtain yeast-like cells . The organism was maintained in the mycelial form, and its transformation to yeast at the higher temperature proceeded via conidia and 'intermediate cells' that then gave rise to yeast by a blastic mechanism . Cell-free extracts were analysed by PAGE at pH 8.0 and acid phosphatases (EC 3.1.3.2) were revealed by a sensitive detection reagent at pH 5.0 . Mycelial, conidial and yeast extracts all had some acid phosphatase activity (M-I, C-I and Y-I) at the origin, although the proportion was highest for the yeast extracts . All of the bands that penetrated the gels had different electrophoretic mobilities . Mycelial and conidial extracts each had one other isoenzyme (M-II and C-II), while the yeast extracts had a total of five electrophoretically distinct acid phosphatases . Isoenzyme Y-II was further resolved into five closely related bands (Y-IIa to Y-IIe), the relative intensities of which varied with the phosphate nutrition of the yeast cells and the history of the extracts . The acid phosphatase isoenzymes were inhibited to various extents by sodium fluoride, L(+)-tartrate and phosphate, and showed interactions with citrate as opposed to acetate as the background buffer at pH 5.0.

Arzneimittelforschung, 1986 Dec, 36(12), 1796 - 800
Pharmacological investigations of the new antiinflammatory agent 2-(10,11-dihydro-10-oxodibenzo{b,f}thiepin-2-yl)propionic acid . 1st communication: inhibitory effects of rat paw edema; Tsurumi K et al.; In the development process of a new nonsteroidal antiinflammatory drug (NSAID) with less toxicity and side-effects, 2-(10,11-dihydro-10-oxodibenzo{b,f}thiepein-2-yl) propionic acid (CN-100) was chosen as the most excellent NSAID from synthetic tricyclic compounds after screening test . For the series of studies on antiinflammatory effects of this compound in detail, its effect on rat paw edema induced by various phlogists was first investigated . The inhibitory effect of CN-100 on carrageenin-induced edema was remarkable and nearly equal to that of indometacin . The effect was not affected by continuous administration for 2 weeks or adrenalectomy . Similarly to indometacin, CN-100 had no significant effect on yeast-induced edema mediated by 5-hydroxytryptamine and concanavalin-A-induced edema unrelated with prostaglandins . However, CN-100 displayed a weaker inhibitory effect on nystatin-induced edema than indometacin, suggesting that CN-100 has a low membrane stabilizing action and a strong blocking action on synthesis of prostaglandins . CN-100 inhibited the sustained edema induced by mustard, but the drug did not interfere with the increase in body weight of rats . Indometacin in the same dose caused decrease in body weight and death . The toxicity of CN-100 was definitely less than that of indometacin, although both drugs were similar in antiinflammatory activity and mode of action on rat paw edema . Results suggest that CN-100 is an effective drug on not only acute but also subacute and chronic inflammation.

J Natl Cancer Inst, 1986 Dec, 77(6), 1281 - 6
Effects of dietary selenium on bis(2-oxopropyl)nitrosamine-induced carcinogenesis in Syrian golden hamsters; Birt DF et al.; In studies designed to determine the influence of dietary Se on pancreatic carcinogenesis, Syrian golden hamsters were fed unsupplemented torula yeast diet or diet supplemented with 0.1 or 5.0 ppm Se, from sodium selenite, starting at 4 weeks of age until the termination of the study . In separate groups, hamsters were given the diet supplemented with 0.1 ppm Se until 5 days after carcinogen treatment . Then they were fed either the unsupplemented diet or the diet supplemented with 5.0 ppm Se until the end of the experiment . N-Nitrosobis(2-oxopropyl)amine (BOP; CAS; 60599-38-4) treatment was given as a single sc injection of 20 mg/kg (body wt) at 8 weeks of age, and surviving hamsters were killed 50 weeks later . As a measure of Se status, glutathione peroxidase (GSHPX) activities were determined in plasma, erythrocytes, and liver . Values were elevated in animals fed higher levels of dietary Se . BOP treatment depressed plasma GSHPX at 24 hours and elevated erythrocyte and liver values at 4 weeks . Pancreatic ductular adenoma yields were inhibited with each elevation of dietary Se in female hamsters fed the diets, both before and after BOP administration, and were further inhibited in females that were fed diets containing 0.1 ppm Se before BOP administration and that were changed to the unsupplemented or 5.0-ppm-supplemented diets after BOP was given . Pancreatic ductular adenoma yields were highest in all male groups given diets of 0.1 ppm Se before BOP administration, irrespective of the Se level after BOP was fed . Adenoma yields in males were lowest in hamsters fed unsupplemented diet, both before and after BOP treatment . Pancreatic carcinoma yields were low and not influenced by dietary Se . The incidence of hepatic necrosis was elevated in BOP-treated hamsters fed the unsupplemented diet, and that of biliary cystic adenomas was highest in the group fed 0.1 ppm Se before and after BOP treatment.

Mol Cell Endocrinol, 1986 Dec, 48(2-3), 111 - 20
Preparation and characterization of submitochondrial fractions from adrenal cells; della-Cioppa G et al.; A method for preparing submitochondrial fractions from adrenocortical cells was developed by adapting a procedure that has been successful with yeast mitochondria . The method is based upon osmotic swelling, sonication and centrifugation in sucrose . The preparation yields highly purified fractions of outer membrane, intermembrane space, inner membrane and a less purified fraction of matrix . Recoveries are good so that 10(7) cells yield approximately 170 micrograms of inner membrane protein and 12 micrograms of outer membrane protein . Electron microscopy shows that the outer membrane fraction consists of vesicles (0.2-0.6 micron diameter) while inner membrane appears as densely packed sheets of membranous material . Two-dimensional polyacrylamide gels (isoelectric focusing followed by electrophoresis) of all the fractions give highly reproducible patterns of protein spots with Coomassie staining . Steroidogenic proteins were found only in inner membrane fractions which were shown to contain cytochrome P-450 C27 side-chain cleavage and P-450 11 beta-hydroxylase together with adrenodoxin and adrenodoxin reductase . Inner membrane catalyzes side-chain cleavage of cholesterol (conversions to pregnenolone) and 11 beta-hydroxylation (DOC----corticosterone) when substrate and NADPH are added . The preparation yields highly purified submitochondrial fractions from Y-1 mouse adrenal tumor cells and from porcine and bovine adrenocortical mitochondria . The method has the virtue of yielding highly purified intermembrane fluid which is not true of other methods for fractionation of adrenal mitochondria . The procedure also yields cleaner preparations of the two membranes than two other published methods currently used to prepare submitochondrial fractions from adrenocortical cells.

Arch Biochem Biophys, 1986 Dec, 251(2), 767 - 70
Enzymatic reactions involving orthoarsenate: arsenate is competitive with sulfate in the ATP sulfurylase reaction; Knowles FC; In the presence of ATP, Mg2+, and arsenate, ATP sulfurylase from yeast will catalyze the formation of inorganic pyrophosphate . Inorganic pyrophosphate was detected by determination of orthophosphate in the presence of inorganic pyrophosphatase . Two moles of Pi were found for each molecule of ATP in the reaction mixture . The activity of ATP sulfurylase with arsenate as an activating anion was from 1 to 3% of the activity observed with molybdate.

Proc Natl Acad Sci U S A, 1986 Dec, 83(24), 9313 - 7
Molecular analysis of cDNA clones and the corresponding genomic coding sequences of the Drosophila dunce+ gene, the structural gene for cAMP phosphodiesterase; Chen CN et al.; We have isolated and sequenced cDNA clones representing portions of the polyadenylylated transcripts of the dunce+ gene . These define an open reading frame of 1086 bases and some of the 5'- and 3'-untranslated regions of the transcripts . The deduced amino acid sequence is strikingly homologous to the amino acid sequence of a Ca2+/calmodulin-dependent cyclic nucleotide phosphodiesterase isolated from bovine brain and more weakly related to the predicted amino acid sequence of a yeast cAMP phosphodiesterase . These homologies, together with prior genetic and biochemical studies, provide unambiguous evidence that dunce+ codes for a phosphodiesterase . In addition, the dunce+ gene product shares a seven-amino acid sequence with a regulatory subunit of cAMP-dependent protein kinase that is predicted to be part of the cAMP binding site . We also identify a weak homology between a region of the dunce+ gene product and the egg-laying hormone precursor of Aplysia californica . The open reading frame is divided in the genome by four introns.

J Am Mosq Control Assoc, 1986 Dec, 2(4), 545 - 7
Laboratory colonization and life cycle of Coquillettidia crassipes in Malaysia; Chiang GL et al.; Methods are described for the laboratory colonization of Coquillettidia crassipes . The highest rate of insemination occurred in 60 x 60 x 120 cm cages and better insemination in laboratory adapted F15 generation . Embryonation and hatchability of eggs ranged from 69.6 to 97.9% and 63.3 to 94.3% respectively . Gravid females laid egg rafts on water in 500 ml breakers with small leaves of Salvinia for resting . Newly hatched larvae were set up in a basal medium of guinea pig dung and water or liver powder, yeast powder and water . Larvae attached to aquatic plants or 'Keaykolour' ruffia snow white paper . The cultures with paper gave better yields . At present 21 generations of Cq . crassipes have been reared in the laboratory.

Infect Immun, 1986 Dec, 54(3), 714 - 22
Development and characterization of Histoplasma capsulatum-reactive murine T-cell lines and clones; Deepe GS Jr et al.; Experimental studies have suggested that antigen-specific T lymphocytes are important mediators of resistance to infection with the pathogenic fungus Histoplasma capsulation . To gain a better understanding of the role of T lymphocytes, we developed murine T-cell lines and clones that recognized Histoplasma antigens . These T cells were of the helper/inducer phenotype (Thy-1.2+ Lyt-1+ L3T4+ Lyt-2-) and exerted multiple immunological functions . T-cell lines and 12 clones proliferated vigorously in response to histoplasmin; the T-cell lines and 6 clones also were reactive with heterologous fungal antigens prepared from either Blastomyces dermatitidis or Coccidioides immitis . Recognition of antigen by T cells was H-2 restricted; in the absence of antigen, four clones demonstrated alloreactivity . All T-cell clones conferred local delayed-type hypersensitivity responses when injected with antigen into footpads of mice . Ten of 12 T-cell clones released interleukin-2 after stimulation with antigen, and all clones tested secreted interferon . Moreover, culture supernatants from antigen-stimulated clones armed peritoneal macrophages to inhibit intracellular growth of H . capsulatum yeast cells . All clones assayed exerted nonspecific help . Thus, development of T-cell clones should facilitate analysis of the regulatory properties of Histoplasma-specific T cells.

J Immunol Methods, 1986 Nov 20, 94(1-2), 209 - 14
A colorimetric method for measuring the candidacidal activity of leucocytes; Ashman RB; The candidacidal activity of mouse neutrophils was determined by a colorimetric method, using a tetrazolium dye that detects living, but not dead, yeast cells . The assay is carried out in microtitre trays, read using a multiwell scanning spectrophotometer (ELISA reader), and can be analysed by computer.

Eur J Biochem, 1986 Nov 17, 161(1), 13 - 8
Molecular cloning of the cDNA for androgen-dependent sperm-coating glycoproteins secreted by the rat epididymis; Brooks DE et al.; cDNA clones coding for two closely related androgen-dependent sperm-coating glycoproteins secreted by the rat epididymis were selected by screening an epididymal cDNA library constructed in lambda gt 11 with affinity-purified antibody directed against the glycoproteins . The largest clone of 956 nucleotides provided coding information for a protein of 246 amino acids of which the first 19 residues comprise a putative signal peptide sequence which when cleaved would produce a mature protein of 227 residues and a molecular mass of 26 kDa . Confirmation of the identity of the clone was provided by a match between the amino acid sequence predicted from the cDNA sequence and the actual amino acid sequence determined for a tryptic peptide fragment of one of the pure glycoproteins . It is probable that the primary amino acid sequence of the two glycoproteins is identical . Northern blot and slot-blot analysis revealed that the mRNA for the glycoproteins is approximately 1250 nucleotides long and that the concentration of the mRNA in the epididymis is androgen-dependent . The glycoproteins and their mRNAs were unique to the epididymis as determined by Western and Northern blots, respectively, since signals were absent from skin, brain, liver, kidney, heart, skeletal muscle and testis . Cross-reacting proteins of slightly smaller apparent molecular mass were detected in extracts of mouse and guinea-pig epididymis, but not rabbit or bull epididymis . Comparison with existing protein data bases revealed that the epididymal glycoproteins display significant sequence homology with yeast carboxypeptidase Y.

Nucleic Acids Res, 1986 Nov 11, 14(21), 8513 - 33
Detection, sequence patterns and function of unusual DNA structures; Anderson JN; Unusual DNA structures were detected by an electrophoretic procedure in which DNA fragments were separated according to size on agarose gels and then by shape on polyacrylamide gels . Fragments from yeast centromeres migrated faster in polyacrylamide than predicted from their base composition and size and this property was attributed to a nonrandom distribution of oligomeric A tracts that exhibited minima at 10-11 base intervals . Fragments from seven loci in 107 kb of DNA migrated anomalously slow and these fragments contained blocks of A2-6 in a 10-11 base periodicity which is indicative of bent DNA . The most pronounced bent sequences were found within yeast ARS1 and centered at 245 and 240 bp from the left and right ends of the adenovirus genome . Each sequence is approximately 150 bp away from a replication origin and the adenovirus sequences are within 50 bp of enhancers . Nuclear matrix attachment sites, which are also adjacent to enhancers, contain sequences characteristic of bent DNA . These results suggest that bent structures reside at the base of DNA loops in chromosomes.

Nippon Yakurigaku Zasshi, 1986 Nov, 88(5), 369 - 74
{Effect of cysteine ethylester hydrochloride (Cystanin) on host defense mechanisms (III): Potentiating effects on phagocytosis and nitroblue tetrazolium (NBT) reduction by leukocytes of mice and guinea pigs}; Hisadome M et al.; ICR mice were treated orally with cysteine ethylester hydrochloride (ethylcysteine, 10 and 100 mg/kg) immediately before the intraperitoneal injection of yeast particles . This agent significantly potentiated phagocytosis of yeast particles by peritoneal polymorphonuclear leukocytes in mice obtained 2 hr after the yeast injection, and the treatment with this agent (3 and 30 mg/kg, p.o.) 4 hr before the injection of yeast potentiated phagocytosis of yeast particles by mouse peritoneal leukocytes . This agent (30 mg/kg, p.o.) restored the suppression of phagocytosis of mouse leukocytes by the intraperitoneal administration of cyclophosphamide (30 mg/kg, i.p.) 24 hr before the yeast injection . This agent (10-100 mg/kg, p.o.) had no effect on the decrease of peripheral leukocyte number in irradiated mice (560 rad), but restored the suppression of phagocytosis, nitroblue tetrazolium (NBT) reduction and stimulated NBT reduction by the addition of lipopolysaccharide . Furthermore, this agent (3-30 mg/kg, p.o.) potentiated phagocytosis, NBT reduction and stimulated NBT reduction of peripheral leukocytes obtained from guinea pigs 2 and 6 hr after ethylcysteine treatment . It is suggested that ethylcysteine potentiates phagocytosis and NBT reduction of leukocytes in animals, and it restores phagocytosis and NBT reduction inhibited by the treatment with cyclophosphamide or X-ray irradiation . It may be possible that this stimulating effect of ethylcysteine could be at least in part involved in the stimulation of nonspecific resistance to infection in the compromised host.

Genetics, 1986 Nov, 114(3), 955 - 70
Mitochondrial DNA transmission genetics in crickets; Rand DM et al.; This paper presents the results of a single generation study of the transmission genetics of mitochondrial DNA in the field cricket Gryllus firmus . In this species, individuals heteroplasmic for at least two different-sized mitochondrial genomes can be collected easily from natural populations . The frequencies of mtDNA size variants in heteroplasmic females and samples of their offspring were estimated by densitometry of autoradiographs . The variance in mitochondrial genotype frequencies among the offspring of heteroplasmic females indicates that, through genetic drift, fixation would take several hundred animal generations . Differences between the observations and data on mtDNA transmission in yeast and cows are discussed in light of the differences in organelle sampling regime and early developmental events in these species . Our data also show shifts in genotype frequencies in the transmission from mother to offspring that suggest a bias in favor of smaller genomes . The nature of mtDNA size variation in natural populations of crickets is discussed in reference to a mutation-selection balance.

Chest, 1986 Nov, 90(5), 656 - 61
Pulmonary alveolar proteinosis . Further evaluation of abnormal alveolar macrophages; Gonzalez-Rothi RJ et al.; To investigate the function of alveolar macrophages (AM) and the mechanisms of impairment in pulmonary alveolar proteinosis, we established in culture AM from three patients and from eight normal nonsmokers and assessed phagocytosis and phagolysosome fusion by the acridine orange assay with live yeast as the phagocytic challenge . Alveolar macrophages from the patients with pulmonary alveolar proteinosis ingested fewer yeasts per cell than did normal AM (mean +/- SE, 2.3 +/- 0.3 vs 3.3 +/- 0.2; p less than 0.05) and had decreased phagolysosome fusion (33 +/- 6 percent vs 64 +/- 1 percent; p less than 0.001) . Alveolar macrophages from three normal subjects were incubated with cell-free fractions isolated by centrifugation of lavage fluid from the patients at 250 g (P1) or centrifugation of P1 supernatant at 20,000 g (P2) . The P1 fraction did not decrease the number of AM ingesting yeast or the number of yeast cells ingested per cell, but the P2 fraction decreased both phagocytic indices . Conversely, phagolysosome fusion was depressed by the P1 fraction (48 +/- 3 percent vs 66 +/- 2 percent for untreated AM from the same subject; p less than 0.02) but not by the P2 fraction . Significant morphologic changes were noted in AM cocultured with both P1 and P2 . Comparable concentrations of pooled P2 fractions from normal subjects did not decrease phagocytic indices in normal AM . These data confirm that AM in pulmonary alveolar proteinosis are dysfunctional, and, in particular, the finding of decreased phagolysosome fusion may be related to the high incidence of uncommon infections in these patients . We have shown that different fractions of alveolar filling material from patients with pulmonary alveolar proteinosis have unique effects on the phagocytic process in the normal AM, and the induced defects may be associated with apparent uptake of this material . These observations further support the hypothesis that in patients with pulmonary alveolar proteinosis, locally produced "toxic" substances may lead to impaired alveolar clearance and contribute to the pathogenesis of this disease.

J Immunol, 1986 Nov 1, 137(9), 2913 - 7
Ribonuclease activity associated with human eosinophil-derived neurotoxin and eosinophil cationic protein; Slifman NR et al.; The eosinophil granule contains a series of basic proteins, including major basic protein, eosinophil peroxidase, eosinophil-derived neurotoxin (EDN), and eosinophil cationic protein (ECP) . Both EDN and ECP are neurotoxins and helminthotoxins . Comparison of the partial N-terminal amino acid sequences of EDN and ECP showed 67% identity; surprisingly, they also showed structural homology to pancreatic ribonuclease (RNase) . Therefore, we determined whether EDN and ECP possess RNase enzymatic activity . By spectrophotometric assay of acid soluble nucleotides formed from yeast RNA, purified EDN showed RNase activity similar to bovine pancreatic RNase, whereas ECP was 50 to 100 times less active . The RNase activity associated with ECP was not significantly inhibited after exposure of ECP to polyclonal or monoclonal antibody to EDN . These results indicate that EDN and ECP both possess RNase activity, the RNase activity of EDN and ECP is specific, and EDN and ECP have maintained not only structural but also functional homology to pancreatic RNase.

Curr Eye Res, 1986 Nov, 5(11), 833 - 40
Immunofluorescent detection of Histoplasma capsulatum in primate experimental ocular histoplasmosis; Miyashiro JE et al.; We have developed a polyclonal anti-Histoplasma capsulatum antibody to detect H . capsulatum antigens in ocular tissue . Antibodies were specific for H . capsulatum as determined by enzyme linked immunosorbent assay and immunoprecipitation . A total of 21 choroidal histoplasmic lesions in 4 primate eyes (Macaca speciosa and Macaca mulatta), taken at various times from 14 to 60 days after the internal carotid artery injection of yeast phase organisms, were evaluated . Using an indirect immunofluorescent technique, these antibodies stained yeast phase organisms within acute choroidal lesions at 14 days after infection . By 60 days intact organisms were no longer detected; occasional cells, however, contained intracellular inclusions that stained with these antibodies . Although yeast phase organisms are rapidly cleared from the primate choroid, these data indicate that residual H . capsulatum antigens may remain in choroidal lesions after the acute infectious stage . These findings are consistent with the hypothesis that H . capsulatum antigens play a role in the immunologic reactivation of atrophic choroidal scars in ocular histoplasmosis.

Am J Prev Med, 1986 Nov-Dec, 2(6), 324 - 9
Endometriosis: a comparison of associated disease histories; Lamb K et al.; We examined a subgroup of 43 women with endometriosis who reported family members affected by the disease . A matched group of female friend-controls (n = 43) was used for comparison, and reports of selected autoimmune diseases, infectious diseases, and allergic manifestations were compared . For this subset of women, there appeared to be a strong familial tendency to allergic manifestations . Vaginal yeast infections, a history of mononucleosis, eczema, hayfever, and food sensitivities were reported to occur much more frequently for these women . All statistical tests on these conditions were significant at the .05 level of probability or less.

Pharmazie, 1986 Nov, 41(11), 796 - 9
{The effect of ambroxol and bromhexine on the "respiratory burst" of alveolar macrophages}; Winsel K et al.; The effect of ambroxol and bromhexine on the yeast cell wall- and arachidonic acid (AA)-induced luminol-respectively lucigenin-dependent chemiluminescence (CL) of alveolar macrophages (AM) of patients with lung diseases has been investigated . Both drugs cause a suppression of the induced CL . These results suggest that ambroxol and bromhexine reduce the generation of reactive oxygen metabolites (ROM) by AM . The mechanism of action is thought to involve the activation of the acyl-CoA: lysophosphatide acyltransferase . The increase of this enzyme activity lowers probably the intracellular concentration of free AA and consequently also the production of ROM . The investigations show a new possibility for the influence of biosynthesis of ROM and likely of eicosanoids, which play an important role as pathogenetic mediators in different lung diseases.

J Bacteriol, 1986 Nov, 168(2), 843 - 50
Effect of L-amino acids on Mucor rouxii dimorphism; Leija A et al.; Mucor rouxii organisms growing aerobically and exponentially on a well-defined minimal medium are able to differentiate as yeasts or as mycelia, depending on the amino acid as the nitrogen source . When certain amino acids were used as the nitrogen source, spores differentiated only as hyphae, whereas other amino acids gave rise to other morphological forms having different ratios of yeasts to hyphae . In both hyphal and yeast cultures, an aerobic metabolism was predominant, as shown by determining several metabolic parameters such as oxygen tension, glucose consumption, ethanol production, and CO2 release . A complete conversion of yeasts to hyphae was obtained by the appropriate change in the amino acid used as nitrogen source . By preparing spheroplasts from mycelial cultures and transferring them to media with amino acids that induce yeast formation, a 50% yield in the reverse transformation was achieved . A correlation between the change in pH of the medium and cell morphology was observed in different growth conditions . Decrease in the pH of the medium preceded the appearance of hyphae . Also, when the initial pH of the medium was increased, aspartate-containing cultures developed mainly as mycelia, instead of yeasts, with a corresponding decrease in the final pH.

J Clin Invest, 1986 Nov, 78(5), 1220 - 8
Recombinant human granulocyte-macrophage colony-stimulating factor stimulates in vitro mature human neutrophil and eosinophil function, surface receptor expression, and survival; Lopez AF et al.; A purified recombinant human granulocyte-macrophage colony stimulating factor (rH GM-CSF) was a powerful stimulator of mature human eosinophils and neutrophils . The purified rH GM-CSF enhanced the cytotoxic activity of neutrophils and eosinophils against antibody-coated targets, stimulated phagocytosis of serum-opsonized yeast by both cell types in a dose-dependent manner, and stimulated neutrophil-mediated iodination in the presence of zymosan . In addition, rH GM-CSF enhanced N-formylmethionylleucylphenylalanine(FMLP)-stimulated degranulation of Cytochalasin B pretreated neutrophils and FMLP-stimulated superoxide production . In contrast, rH GM-CSF did not promote adherence of granulocytes to endothelial cells or plastic surfaces . rH GM-CSF selectively enhanced the surface expression of granulocyte functional antigens 1 and 2, and the Mo1 antigen . rH GM-CSF induced morphological changes and enhanced the survival of both neutrophils and eosinophils by 6 and 9 h, respectively . These experiments show that granulocyte-macrophage colony stimulating factor can selectively stimulate mature granulocyte function.

Science, 1986 Oct 31, 234(4776), 614 - 8
URF6, last unidentified reading frame of human mtDNA, codes for an NADH dehydrogenase subunit; Chomyn A et al.; The polypeptide encoded in URF6, the last unassigned reading frame of human mitochondrial DNA, has been identified with antibodies to peptides predicted from the DNA sequence . Antibodies prepared against highly purified respiratory chain NADH dehydrogenase from beef heart or against the cytoplasmically synthesized 49-kilodalton iron-sulfur subunit isolated from this enzyme complex, when added to a deoxycholate or a Triton X-100 mitochondrial lysate of HeLa cells, specifically precipitated the URF6 product together with the six other URF products previously identified as subunits of NADH dehydrogenase . These results strongly point to the URF6 product as being another subunit of this enzyme complex . Thus, almost 60% of the protein coding capacity of mammalian mitochondrial DNA is utilized for the assembly of the first enzyme complex of the respiratory chain . The absence of such information in yeast mitochondrial DNA dramatizes the variability in gene content of different mitochondrial genomes.

Nucleic Acids Res, 1986 Oct 24, 14(20), 8111 - 9
Small tandemly repeated DNA sequences of higher plants likely originate from a tRNA gene ancestor; Benslimane AA et al.; Several monomers (177 bp) of a tandemly arranged repetitive nuclear DNA sequence of Brassica oleracea have been cloned and sequenced . They share up to 95% homology between one another and up to 80% with other satellite DNA sequences of Cruciferae, suggesting a common ancestor . Both strands of these monomers show more than 50% homology with many tRNA genes; the best homologies have been obtained with Lys and His yeast mitochondrial tRNA genes (respectively 64% and 60%) . These results suggest that small tandemly repeated DNA sequences of plants may have evolved from a tRNA gene ancestor . These tandem repeats have probably arisen via a process involving reverse transcription of polymerase III RNA intermediates, as is the case for interspersed DNA sequences of mammalians . A model is proposed to explain the formation of such small tandemly repeated DNA sequences.

Nucleic Acids Res, 1986 Oct 24, 14(20), 7995 - 8006
Sequence and transcription analysis of the Petunia mitochondrial gene for the ATP synthase proteolipid subunit; Young EG et al.; We have sequenced the Petunia hybrida gene that specifies the proteolipid subunit of the mitochondrial Fo ATP synthase and have used this gene to investigate plant mitochondrial gene transcription . The Petunia atp 9 gene contains a single open-reading frame capable of specifying a 77 amino acid-polypeptide that is homologous to bovine, fungal and maize proteolipid subunits . S1 protection identified 3 transcripts in a ratio of 1:5:100 in the Petunia tissues tested . The transcripts share a common 3' terminus but have 5' termini that map 528, 266, and 121 nucleotides upstream of the translation start site . The 5' terminus of the longest transcript maps to the sequence ATATAGTA, which is nearly identical to the yeast mitochondrial transcription initiation site ATATAAGTA . Primer extension analysis indicates that these two shorter transcripts are not due to splicing . The two shorter transcripts originate at sequences homologous to sites at 5' termini of two pea and maize genes . These consensus sequences may signal processing events other than splicing.

Eur J Biochem, 1986 Oct 15, 160(2), 343 - 8
Simple-kinetic descriptions of alcohol dehydrogenase after immobilization on tresyl-chloride-activated agarose; Bille V et al.; Yeast alcohol dehydrogenase was successfully immobilized on tresyl-chloride-activated agarose; the optimized conditions allowed an enzyme activity recovery of over 90% . Comparison of free and immobilized enzyme properties showed an unchanged intrinsic activation energy of the reaction and a shift of optimum activity to a higher pH medium after immobilization . Comparison of the kinetic parameters for both substrates of the reaction showed that the Michaelis-Menten model could not take into consideration all the constraints induced by the immobilization on the enzyme properties but that the Theorell-Chance model was more appropriate . These results are discussed taking into consideration the factors affecting the immobilized enzyme . Finally, we discuss the possibilities of cofactor regeneration with this immobilized alcohol dehydrogenase.

Schweiz Med Wochenschr, 1986 Oct 11, 116(41), 1401 - 10
{Food allergies . III . Therapy: elimination diet, symptomatic drug prophylaxis and specific hyposensitization}; Wuthrich B et al.; The treatment of food allergies is logically based on strict elimination of causative allergens . While it is easy to eliminate food which is infrequently consumed, it is more difficult to manage an allergy involving regularly consumed foods, especially where patients have to eat away from home for professional reasons . The creation of elimination diets for milk, eggs, and mould and yeast allergies is discussed . In raw food and vegetable allergy the act of cooking is often sufficient to denature the allergen as it is unstable to heat . Follow-up investigations show that some 50% of children achieve cure spontaneously by strict elimination diet, especially in regard to milk allergy . In our own 173 (mainly adult) patients with food allergy, some 2/3 reported after 3-5 years that a strict elimination diet had to be followed, since otherwise prompt relapse of allergic symptoms was noted . About 1/3 of patients, mainly with milk, cheese or egg allergy, can hope for spontaneous desensitization by appropriate diet . This is demonstrated by a case history with disappearance of IgE antibodies . Should this fail to occur, oral desensitization with milk or egg-white extracts offers an effective therapy . The practice of hyposensitization with foodstuffs is illustrated by examples and tabulation of immunologic parameters . In raw food or vegetable allergy, which is often associated with birch or mugwort pollinosis, improvement or even complete cure can be expected in about 1/3 of cases by systematic desensitization of pollinosis . On the other hand, the therapy and prognosis of food allergy involving extreme and polyvalent sensitivities, especially to spices, or with multifactorially induced symptoms, is more problematic . In these cases a strict elimination diet should be followed by continuous prophylactic/symptomatic treatment with antianaphylactic substances such as cromoglicinic acid (Nalcrom) - especially in gastrointestinal food allergies - or with ketotifen (Zaditen) or oxatomide (Tinset) in hematogenically released shock fragments . Patients with severe anaphylactic reactions after meals should, by analogy with patients with hymenoptera allergy, carry an emergency kit containing an adrenaline spray (Medihaler-Epi), a soluble corticosteroid (e.g . Betnesol) and anthistaminic drugs (e.g . Tavegyl, Teldane).

Biochemistry, 1986 Oct 7, 25(20), 6244 - 51
Amino acid sequence of a chicken heat shock protein derived from the complementary DNA nucleotide sequence; Kulomaa MS et al.; The complete nucleotide sequence for a chicken heat shock protein (hsp108) was determined from cDNA clones isolated from hen oviduct and bursal lymphoma recombinant DNA libraries . This protein has certain biochemical similarities to the progesterone receptor, but it is clearly distinct from it . The initial cDNA clone, isolated from a chicken oviduct cDNA library, was detected by antibody screening and hybrid-selected translation {Zarucki-Schulz, T., Kulomaa, M . S., Headon, D . R., Weigel, N . L., Baez, M., Edwards, D . P., McGuire, W . L., Schrader, W . T., & O'Malley, B . W . (1984) Proc . Natl . Acad . Sci . U.S.A . 81, 6358-6362} . The earlier clones were used to screen for additional cDNAs, and cDNAs that define the entire mRNA sequence of hsp108 have been obtained . The nucleotide sequence codes for peptides present in hsp108 as determined by protein microsequencing . The 5' end of the mRNA was determined by primer extension studies . The mRNA contains a noncoding region of 101 nucleotides upstream from the predicted initiation codon . The 3' untranslated region contains 244 nucleotides beyond the termination codon, and it contains a predicted polyadenylation signal 26 nucleotides from the end of the complete cDNA . The coding region of 2385 nucleotides corresponds to a polypeptide chain of 795 amino acids, giving a molecular weight of 91,555 for the hsp108 protein . In another paper, evidence is presented that hsp108 shows a high degree of amino acid sequence homology with two heat shock proteins, hsp90 (yeast) and hsp83 (Drosophila), and is indeed inducible by heat shock {Sargan, D . R., Tsai, M.-J., & O'Malley, B . W . (1986) Biochemistry (following paper in this issue)}.

Anal Biochem, 1986 Oct, 158(1), 104 - 10
Specifically increased solubility of enzymes in polyethyleneglycol solutions using polymer-bound triazine dyes; Johansson G et al.; The enzymes glucose-6-phosphate dehydrogenase (EC 1.1.1.49) and 3-phosphoglycerate kinase (EC 2.7.2.3), present in an extract of Bakers' yeast, are largely kept in solution by minor amounts of polyethylene glycol-bound triazine dyes (Procion yellow HE-3G and Procion olive MX-3G) even when the solution contains such concentrations of polyethylene glycol (12.5% w/w) which normally precipitate the enzymes . The specific prevention from precipitation can be used for purification of enzyme, preferentially in dealing with crude extracts, which has been demonstrated in this work . A 3.4-fold purification of glucose-6-phosphate dehydrogenase has been achieved with good recovery (93%) . Further purification has been possible by combining the recovered (enzyme-containing) supernatant liquid with a solution of dextran which generates an aqueous two-phase system . The lower, dextran-containing phase extracts part of the remaining bulk proteins leaving the target enzyme in the upper phase . The advantages of this method for enzyme purification in large scale are discussed.

J Med Vet Mycol, 1986 Oct, 24(5), 395 - 400
Sarcinomyces phaeomuriformis: a new dematiaceous hyphomycete; Matsumoto T et al.; Sarcinomyces phaeomuriformis is described as a new species in the genus Sarcinomyces Lindner . Currently, the taxon is known to occur only in Japan as a causal agent of phaeohyphomycosis . The colonies are initially yeast-like, which on aging become dry, granular, heaped, friable and black . The initial growth consists of single cells which form multiple broad-based buds and by a successive budding process produce chains of blastoconidia . In its blastic conidiogenesis, S . phaeomuriformis resembles Phaeococcomyces catenatus . However, mature colonies consist of thick-walled, pale to dark brown muriform cells which develop broad-based buds . The buds, after separating from their parent cells, either bud or enlarge and divide internally by septations laid down in different planes to become muriform . Conidiogenesis that gives rise to multiple, broad-based blastic and thallic-sarcinic conidia characterizes S . phaeomuriformis.

Biokhimiia, 1986 Oct, 51(10), 1688 - 95
{Isolation and various properties of soluble and membrane-bound acid RNAse from rat brain lysosomes}; Nechaeva GA; Preparations of soluble (I) and membrane-bound (II) acid RNAse with Mr 68,000 and 72,000 Da, respectively, and purified about 2000-fold were isolated from lysosome-rich fractions of rat brain large hemispheres . RNAase II differed from RNAase I by a lower temperature stability . The pH optimum (pH 5.8-6.1), temperature optimum and substrate specificity of RNAase I and II appeared to be identical . The Km values of RNAases I and II for poly(U) are 166 and 160 micrograms/ml; those for RNA--1200 and 1250 mu k/ml, respectively . RNAases I and II extensively hydrolyze soluble, polymeric RNA, rRNA from brain and yeast and poly(U) but do not influence poly(C), poly(A), poly(G), tRNA and DNA . Monovalent cations (K+, Na+, NH4+) activate both RNAase forms.

Mol Biochem Parasitol, 1986 Oct, 21(1), 83 - 92
Membrane potential of erythrocytic stages of Plasmodium chabaudi free of the host cell membrane; Mikkelsen RB et al.; Free parasites were isolated from Plasmodium chabaudi-infected rat erythrocytes by N2-cavitation and purified on Percoll gradients . The membrane potential of the free parasites determined from the transmembrane distribution of the lipophilic cation, tetraphenylphosphonium, was -93 +/- 10 mV for late stage parasites and -90 +/- 3 mV for ring forms . Studies with intact infected erythrocytes demonstrated that the membrane potential of ring forms was much smaller compared to late trophozoites and schizonts and thus the present findings with free parasites suggest that host cell cytoplasmic factors may determine the magnitude of the parasite membrane potential . Both extracellular pH and {Na+} were found to modify the membrane potential of free parasites . Electrogenic protonophores, the H+-ATPase inhibitor dicyclohexylcarbodiimide and orthovanadate collapsed the potential of free parasites . Ouabain (or its membrane permeant derivative, strophanthidin), and oligomycin were without effect . These inhibitor studies suggest that an electrogenic H+-ATPase similar to that found in yeast generates in part the membrane potential of malaria parasites . Using weak acid distribution or a pH sensitive fluorescent dye, it was demonstrated that free parasites maintain an alkaline intracellular pH at extracellular pH greater than 6.5 . The pH gradient was partially collapsed by orthovanadate or dicyclohexylcarbodiimide and by substitution of Na+ for K+ in the suspending buffer . The H+-ATPase and K+:H+ exchange may therefore both contribute to regulation of intracellular pH in Plasmodium.

Am Rev Respir Dis, 1986 Oct, 134(4), 771 - 6
The pathogenesis of experimental pulmonary histoplasmosis . Correlative studies of histopathology, bronchoalveolar lavage, and respiratory function; Baughman RP et al.; A murine model of acute pulmonary histoplasmosis was employed to study the pathogenesis of the disease process by means of histopathology, bronchoalveolar lavage, and respiratory function tests . These studies were performed on C57BL/6 mice from 8 h to 8 wk after intranasal inoculation of 10(5) yeast forms of Histoplasma capsulatum and on age-matched control animals that received saline only . At Week 1, the histopathology was characterized by subacute inflammation consisting of polymorphonuclear leukocytes (PMN), lymphocytes, and macrophages that infiltrated the interstitium around small bronchioles and adjacent alveoli . At Weeks 2 and 4, the infiltrates were comprised predominantly of lymphocytes and macrophages; noncaseating granulomas were present at Week 2 . Aggregates of lymphoid cells were prominent along the bronchial tree and in perivascular distribution . Those in close contact with bronchiolar epithelium resembled hyperplastic bronchus associated lymphoid tissue . Quantitative studies of cells in the BAL fluid revealed a large influx of PMN at Week 1 with return to normal range by Week 2 . At this time there was a significant (p less than 0.02) increase in lymphocytes that persisted through Week 8, although histopathologic changes were minimal in lung at this time . A significant decrease in the DLCO/TLC at Week 2 in association with a normal vital capacity indicated impairment of respiratory function secondary to the alveolitis induced by H . capsulatum infection rather than a reduction of lung volume . This model offers promise for additional correlative studies of lymphocyte subsets in lung tissue and alveolar spaces as well as of the functions subserved by these respective populations.

Arch Otolaryngol Head Neck Surg, 1986 Oct, 112(10), 1090 - 2
Oropharynx decontamination preventing Candida vegetation on voice prostheses; Mahieu HF et al.; Candida vegetations found in silicone voice prostheses used in postlaryngectomy voice rehabilitation are associated with prosthesis dysfunction . Selective oropharyngeal decontamination with amphotericin B lozenges resulted in sharp decrease of oropharyngeal Candida concentrations with a subsequent reduction of yeast colonization on voice prostheses and in tracheoesophageal fistula in ten patients . A case report suggests that this selective oropharyngeal decontamination can prolong the device's lifetime.

Proc Natl Acad Sci U S A, 1986 Oct, 83(19), 7211 - 5
Cloning and sequence analysis of cDNA for human argininosuccinate lyase; O'Brien WE et al.; Using antibodies specific for argininosuccinate lyase (EC 4.3.2.1), we isolated two cDNA clones by screening a human liver cDNA library constructed in the lambda gt11 expression vector . The identity of these isolates was confirmed by in vitro translation of plasmid-selected mRNA . One of these isolates was used to rescreen the cDNA library and a 1565-base-pair (bp) clone was identified . The entire nucleotide sequence of this clone was determined . An open reading frame was identified which encoded a protein of 463 amino acids with a predicted molecular weight of 51,663 . The clone included 115 bp of 5' untranslated sequence and 46 bp of 3' untranslated sequence . A canonical poly(A) addition site was present in the 3' end, 16 bp from the beginning of the poly(A) tract . Comparison of the deduced amino acid sequence of the human enzyme with that of the yeast enzyme revealed a 56% homology, when conservative amino acid changes were taken into consideration . The yeast protein is also 463 amino acids long, with a molecular weight of 51,944 . By use of a genomic DNA panel from human-Chinese hamster somatic cell hybrids, the human gene was mapped to chromosome 7 . Another hybridizing region, corresponding to a portion of the 5' end of the cDNA, was found on chromosome 22.

Chem Biol Interact, 1986 Oct 1, 59(3), 295 - 300
Increased proliferative activity in selenium-deficient mouse liver; Otter R et al.; Male albino NMRI mice were fed a selenium-deficient (Se-), torula yeast-based diet containing less than 10 ppb Se for at least 2 months (Se-) while a control group received the same diet supplemented (Se+) with 330 ppb Se as Na2SeO3 . The Se-(-)animals showed multiple enzyme modulations of liver enzyme activities indicating that they were in a severely Se- state . No significant difference in the basic DNA synthesis rate of Se-(-)animals compared to Se+ controls was measured . However, when liver cell proliferation was induced by either hepatopoietin pretreatment or by partial hepatectomy, an about 3-fold increase in DNA replication rates was found in Se- compared to controls . We conclude that the enhanced proliferative activity in Se- mouse liver is expressed in an emergency situation.

Eur J Biochem, 1986 Sep 15, 159(3), 493 - 7
Sulfite oxidase from chicken liver . The role of imidazole and carboxyl groups for the reaction with cytochrome c; Ritzmann M et al.; Oxidation of sulfite to sulfate by sulfite oxidase is inhibited when the enzyme is treated with reagents known to modify imidazole and carboxyl groups . Modification inhibits the oxidation of sulfite by the physiological electron acceptor cytochrome c, but not by the artificial acceptor ferricyanide . This indicates interference with reaction steps that follow the oxidation of sulfite by the enzyme's molybdenum cofactor . Reaction with diethylpyrocarbonate modifies ten histidines per enzyme monomer . Loss of activity is concomitant to the modification of only a single histidine residue . Inactivation takes place at the same rate in free sulfite oxidase and in the sulfite-oxidase--cytochrome-c complex . Blocking of carboxyl groups with water-soluble carbodiimides inactivates the enzyme . But none of the enzyme's carboxyl groups seems to be essential in the sense that its modification fully abolishes activity . The pattern of inactivation by chemical modification of sulfite oxidase is quite similar to that observed previously for cytochrome c peroxidase from yeast {Bosshard, H . R., Banziger, J., Hasler, T . and Poulos, T . L . (1984) J . Biol . Chem . 259, 5683-5690; Bechtold, R . and Bosshard, H . R . (1985) J . Biol . Chem . 260, 5191-5200} . The two enzymes have very different structures yet share cytochrome c as a common substrate of which they recognize the same electron-transfer domain around the exposed heme edge.

J Biol Chem, 1986 Sep 15, 261(26), 12209 - 12
The stereospecificity of oxidation of alpha-{4R-2H}NADH by dehydrogenases; Oppenheimer NJ; The stereospecificity of the enzyme-dependent oxidation of alpha-{4R-2H}NADH has been determined for four dehydrogenases: two pro-R specific enzymes, pig heart malate dehydrogenase and yeast alcohol dehydrogenase; and two pro-S specific enzymes, rabbit muscle glycerol-3-phosphate dehydrogenase and Rhodopseudomonas spheroides 3-hydroxybutyrate dehydrogenase . In all cases, an enzyme-dependent and substrate-specific oxidation to alpha-NAD+ is observed with the stereochemistry of oxidation identical with that found for the oxidation of the correspondingly labeled beta-NADH . The ability of dehydrogenases from diverse sources to utilize alpha-NADH in a stereochemically competent fashion is discussed in relation to proposed interactions between the nicotinamide sugar moiety and active site residues or obligatory alignments of the pyridine and sugar moieties.

Cancer Lett, 1986 Sep, 32(3), 323 - 6
Effects of CCNU on monocyte phagocytosis; Athlin L et al.; The effect of CCNU on human monocyte phagocytosis of yeast cells was measured in vitro . 'Therapeutic' CCNU concentrations had no measurable effect on monocyte phagocytosis . Higher concentrations of CCNU had a slight but statistically significant effect on the engulfment phase of monocyte phagocytosis . The results suggest that CCNU treatment is of minor importance for monocyte function.

J Nutr, 1986 Sep, 116(9), 1701 - 10
Deposition of dietary organic and inorganic selenium in rat erythrocyte proteins; Beilstein MA et al.; The deposition of selenium (Se) in erythrocyte proteins was studied in rats fed Se as sodium selenite, selenocystine, selenomethionine (Se-Met), high Se wheat or selenium-enriched yeast . Ion-exchange chromatography of acid hydrolyzates of selenium-enriched yeast, high Se wheat and intrinsically labeled 75Se wheat indicated that the majority of the Se was present as Se-Met . Gel filtration (Sephadex G-150) of erythrocyte lysates revealed that Se was deposited mainly in two proteins, glutathione peroxidase (GPx) and hemoglobin (Hb) . When selenite or selenocystine was the dietary form of Se, the majority of the erythrocyte Se was present with GPx, but when the Se was supplied from either Se-Met, yeast or wheat, it was deposited to a greater extent in Hb than GPx . Injection of 75Se as either Se-Met or selenite gave results consistent with the feeding studies . 75Se-labeled selenite injection resulted in labeling of primarily GPx, but 75Se-Met injection labeled predominantly Hb . Hence, the dietary forms of Se influence the relative distribution of Se between GPx and Hb in erythrocytes, and this may be a factor contributing to the difference between human and animal erythrocytes.

Proc Natl Acad Sci U S A, 1986 Sep, 83(18), 6741 - 5
Molecular cloning and nucleotide sequence of a full-length cDNA for human alpha enolase; Giallongo A et al.; We previously purified a 48-kDa protein (p48) that specifically reacts with an antiserum directed against the 12 carboxyl-terminal amino acids of the c-myc gene product . Using an antiserum directed against the purified p48, we have cloned a cDNA from a human expression library . This cDNA hybrid-selects an mRNA that translates to a 48-kDa protein that specifically reacts with anti-p48 serum . We have isolated a full-length cDNA that encodes p48 and spans 1755 bases . The coding region is 1299 bases long; 94 bases are 5' noncoding and 359 bases are 3' noncoding . The cDNA encodes a 433 amino acid protein that is 67% homologous to yeast enolase and 94% homologous to the rat non-neuronal enolase . The purified protein has been shown to have enolase activity and has been identified to be of the alpha type by isoenzyme analysis . The transcriptional regulation of enolase expression in response to mitogenic stimulation of peripheral blood lymphocytes and in response to heat shock is also discussed.

Acta Cytol, 1986 Sep-Oct, 30(5), 477 - 80
Cytodiagnosis of Candida organisms in cervical smears; Siapco BJ et al.; Cervical smears and cervical scrapings cultured on Sabouraud agar from 31 women suspected of having Candida genital infections were examined in a study of the cytomorphology of this fungal infection in cervical smears . Of the 31 samples, 20 (64.5%) grew C . albicans in culture . One sample (3.2%) grew C . paratropicalis, 2 (6.4%) grew mixed C . albicans and Torulopsis glabrata and 2 (6.4%) grew T . glabrata alone . Of the 25 fungus-positive samples, 20 (80%) had fungus-positive cervical smears and 5 (20%) had fungus-negative smears . There was no instance in which the cervical smear was positive but the culture was negative . Among the cases positive for C . albicans, organisms occurred in two forms: pseudohyphae without blastospores (29.4%) and pseudohyphae with blastospores (70.6%) . T . glabrata was present in the smears as budding and nonbudding yeasts . Although the sensitivity of the cervical smear in detecting fungus in culture-positive patients was only 80%, the cervical smear can still be a useful means of rapid identification of C . albicans when blastospores and pseudomycelium are present . The presence of budding or nonbudding yeast without pseudohyphae should strongly suggest a T . glabrata infection.

Mycopathologia, 1986 Sep, 95(3), 175 - 81
Effects of antineoplastic agents on growth, morphology and metabolism of Torulopsis glabrata; Ghannoum MA; The effect of treatment by a number of antineoplastic agents on the growth, ultrastructure and macromolecular synthesis of T . glabrata was studied . Many differences were noted in the response of this yeast to these agents . Thiotepa and methotrexate inhibited the growth of T . glabrata, while it was resistant to endoxan-asta and vincristine sulphate . A variation of morphological response of T . glabrata was also observed . Methotrexate enhanced filamentation while thiotepa influenced the surface structures of the cells, resulting in loss of cytoplasmic materials and cell collapse . The other two drugs had little or no effect on the morphology of the yeast tested . The incorporation of precursors for macromolecular synthesis of T . glabrata in the presence of thiotepa and methotrexate was restricted . Thiotepa affected the uptake of precursors of RNA, DNA and protein limiting them to between 62 and 66% of the control values . In contrast, methotrexate limited the uptake of macromolecular precursors to a lesser extent . The possible mechanism of action of antineoplastic agents against yeast and the clinical implications of these findings are discussed.

Nature, 1986 Sep 25-Oct 1, 323(6086), 340 - 3
Phenotypic changes induced by a mutated ras gene during the development of Dictyostelium transformants; Reymond CD et al.; The ras proto-oncogene, found in all eukaryotes so far examined, encode s a protein with guanine nucleotide-binding and GTPase activity . Gene disruption experiments in yeast indicate that ras is essential for cell growth . Anit-sense mutagenesis approaches suggest that this is also true for Dictyostelium . Most mutations causing an amino-acid substitution for Gly 12 result in decreased GTPase activity and produce a transforming phenotype . In yeast, a Gly 19---- Val 19, missense mutation (Gly 19 is similar to Gly 12 in mammalian and Dictyostelium ras proteins) causes a series of dominant phenotypes, including elevated adenylate cyclase activity . In mammalian cells there is no evidence that ras activates adenylate cyclase activity . D . discoideum contains a single ras gene (Dd-ras) that encodes a protein very similar to the mammalian ras protein and identical to c-ras at the potentially transforming positions . Dd-ras is expressed in vegetative cells and later in development in prestalk cells whereas ras protein is found in vegetative and developing cells . In the migrating pseudoplasmodium, ras protein is found in prestalk but not prespore cells, suggesting it is involved in the function and/or differentiation of the anteriorly localized prestalk cells . In this report we examine the effects of expression of a Dd-ras gene carrying a Gly-12----Thr 12 missense mutation.

J Nutr, 1986 Sep, 116(9), 1726 - 34
Selenium-dependent glutathione peroxidase inhibitors increase toxicity of prooxidant compounds in chicks; Mercurio SD et al.; The acute lethality of paraquat (1, 1'-dimethyl-4,4'-bipyridinium dichloride; also methyl viologen) for chicks was reduced in a dose-dependent manner by adding to a selenium-deficient torula-yeast-based diet low concentrations (0.02-0.04 ppm) of selenium (Se) as either Na2SeO3, selenomethionine or a high Se yeast without significantly increasing plasma Se-dependent glutathione peroxidase (Se GSH-Px) activity . Similarly, chicks orally dosed with 100 mg nitrofurantoin {N-(5-nitro-2 furfurylidine)-1-aminohydantoin} per kilogram had highest mortalities in the Se-deficient (unsupplemented) group; lowest mortalities occurred in chicks supplemented with 0.2 ppm Se; chicks supplemented with 0.02 ppm Se survived at rates not statistically different from chicks either unsupplemented or supplemented with 0.2 ppm Se . The activities of SeGSH-px in various vital organs were significantly elevated by supplementation of 0.2 ppm Se to Se-deficient chicks; but only kidney SeGSH-Px increased with 0.02 ppm Se . Additionally, no histopathology was observed in the vital organs of moribund chicks 5 or 24 h following nitrofurantoin administration at any dietary level of Se tested . Exposure of chicks to oxygen enhanced the toxicity of nitrofurantoin, but the protective effect of dietary Se was still evident . Two inhibitors of SeGSH-Px, D(-)-penicillamine X HCl and aurothioglucose, were found to increase the lethalities of both nitrofurantoin and paraquat . Aurothioglucose was most effective when administered simultaneously with the prooxidant compounds; penicillamine increased toxicities only when administered at least 24 h before paraquat or nitrofurantoin (it decreased nitrofurantoin lethality and did not significantly alter paraquat toxicity if given simultaneously) . These data support an hypothesis that the protection offered by dietary Se against the acute toxicities of the prooxidant compounds paraquat and nitrofurantoin may be provided by SeGSH-Px in the chick.

Proc Natl Acad Sci U S A, 1986 Sep, 83(18), 7034 - 8
Abnormal response to DNA crosslinking agents of Fanconi anemia fibroblasts can be corrected by transfection with normal human DNA; Diatloff-Zito C et al.; Primary skin fibroblast cell lines from patients with Fanconi anemia were cotransfected with UV-irradiated pSV2neo plasmids and high molecular weight DNA from normal human cells . Restoration of a normal cellular resistance to mitomycin C (MMC) was observed provided that a Fanconi anemia cell line is selected for DNA-mediated transformation (neo gene) and that at least two successive rounds of transfection are performed . Cells were selected by taking advantage of the higher proliferation rate and plating efficiency of the MMC resistant transformants . As estimated from reconstruction experiments, the frequency of transfer of MMC resistance lies between 1 and 30 X 10(-7) . The MMC resistance phenotype was maintained for at least 10 generations following transfection . Evidence for DNA-mediated transformation also includes the recovery of a normal pattern of DNA semiconservative synthesis after treatment with 8-methoxypsoralen and 365-nm UV irradiation, and the presence of exogenous pSV2neo DNA sequences was shown by Southern blot analysis . The acquired MMC resistance is probably due to the presence of DNA from normal cells . Indeed, sensitivity to MMC was maintained when Fanconi anemia cells were cotransfected with the UV-irradiated pSV2neo plasmid mixed with their own DNA or with yeast or salmon sperm DNA . These negative results also render unlikely the selection of spontaneous MMC resistant revertants in transfection of Fanconi anemia cells with normal DNA . These experiments establish the prerequisites for the isolation of the gene(s) involved in the response to DNA crosslinking lesions in human cells.

EMBO J, 1986 Sep, 5(9), 2203 - 8
The ral gene: a new ras related gene isolated by the use of a synthetic probe; Chardin P et al.; We synthesized a set of 20-mer oligonucleotides corresponding to a sequence of seven amino acids strictly conserved in all the different ras proteins, from yeast to man, as well as in rho and YPT, two proteins distantly related to p21 ras (approximately 30% amino acid homology) . This oligonucleotide probe was used to search for new members of the ras family . We describe here a new ras related gene named ral, isolated from a cDNA library of immortalized simian B-lymphocytes . The ral gene codes for a 206 amino acid protein of expected mol . wt 23.5 kd that shares greater than 50% homology with H-ras, K-ras or N-ras . The GTP binding regions of p21 ras and a C-terminal cysteine involved in membrane anchoring are also present in ral; this strongly suggests that ral is a GTP binding protein with membrane localization . Furthermore, several external regions of p21 ras presumably involved in the interaction with effector, receptor and/or regulatory proteins are highly homologous to the corresponding regions in ral . Therefore some of the proteins that interact with ral might be identical or closely related to those interacting with p21 ras.

Anal Biochem, 1986 Sep, 157(2), 385 - 95
ATP sulfurylase-dependent assays for inorganic pyrophosphate: applications to determining the equilibrium constant and reverse direction kinetics of the pyrophosphatase reaction, magnesium binding to orthophosphate, and unknown concentrations of pyrophosphate; Daley LA et al.; A continuous, coupled, spectrophotometric assay is described in which the enzyme ATP sulfurylase is employed to measure the concentration of inorganic pyrophosphate (PPi) at equilibrium with known concentrations of inorganic orthophosphate (Pi) in the presence of excess inorganic pyrophosphatase (PPitase) . In agreement with previous reports, the apparent equilibrium constant (Keq,app) of the PPi hydrolysis reaction was shown to decrease as the concentration of Mg2+ is increased . At pH 7.3, 30 degrees C, in the presence of 150 mM NaCl and 1 mM free Mg2+, Keq,app (calculated as {Pi}t2/{PPi}t) was 1950 . Measurements of Keq,app at different total concentrations of Mg2+ and Pi permitted the determination of K0, the dissociation constant of the Mg-Pi complex . In 0.05 M Tris-Cl, pH 8.0, at 30 degrees C, K0 was 3.6 mM . In the presence of excess ATP sulfurylase, yeast PPitase catalyzed PPi formation from Pi with a specific activity (Vmax) of 9 units X mg protein-1 at pH 8.0, 30 degrees C, and 1 mM free Mg2+ . Half-maximum reverse reaction velocity was observed at a total Pi concentration of 18 mM . (Under the same conditions, Vmax of the PPi hydrolysis reaction was 530 units X mg protein-1.) A radiochemical end point ("reaction-to-completion") assay for measuring unknown concentrations of PPi was devised . In the presence of excess 35S-adenosine-5'-phosphosulfate ({35S}APS) as the cosubstrate, 35SO2-4 formation was stoichiometric with added PPi . (The 35SO2-4 and {35S}APS are separated by adsorption of the latter onto charcoal.) The sensitivity of the assay can be adjusted by varying the specific radioactivity of the {35S}APS . In the absence of interfering substances, as little as 2 pmol of PPi per 1.0 ml assay volume can be measured . The sensitivity of the assay is reduced in the presence of ATP plus perchlorate (which synergistically inhibit the enzyme) . However, if the bulk of the ATP is removed from perchloric acid extracts of tissues with glucose and hexokinase, initial intracellular levels as low as 1 microM can be measured . The possibility that most of the cellular PPi extracted with perchloric acid was originally enzyme bound is discussed.

Vet Immunol Immunopathol, 1986 Sep, 13(1-2), 85 - 95
Cultured bovine monocytes exhibit decreased release of superoxide anion and increased levels of lysosomal enzymes but do not secrete detectable lysozyme activity; Zurbrick BG et al.; In this study we determined the effects of in vitro maturation on the phagocytic activity, lysosomal enzyme content and oxidative response of bovine monocytes . Intracellular levels of the lysosomal enzymes acid phosphatase, beta glucuronidase, and beta glucosaminidase increased as bovine monocytes matured in vitro . However, in marked contrast to the mononuclear phagocytes of other mammalian species, lysozyme activity was undetectable in the culture supernatants and cell lysates of adherent bovine blood monocytes cultured for one to fifteen days . In vitro maturation of bovine monocytes also increased their phagocytic activity as determined by the ingestion of opsonized yeast . A greater percentage of monocyte-derived macrophages that were stimulated with opsonized yeast and phorbol myristic acetate (PMA) reduced nitroblue tetrazolium than did similarly treated monocytes . Monocyte-derived macrophages stimulated with PMA released significantly less superoxide anion than did PMA-stimulated monocytes . Bovine monocytes and macrophages also failed to bind the monoclonal antibody Mac-1, which binds to human and mouse macrophages . Bovine monocytes demonstrated both similarities and differences with other mammalian mononuclear phagocytes, thus making them a useful model for further study of the comparative and developmental biology of mononuclear phagocytes.

Rev Esp Fisiol, 1986 Sep, 42(3), 301 - 8
Evidence for common antigens on human non-adherent synoviocytes (type A) and peripheral monocytes; Perez-Maceda B et al.; The characterization of a homogeneous non-adherent synoviocyte (Type A) cell population (greater than or equal to 95%) from non-rheumatoid patients by culturing the cells in the presence of forty percent foetal calf serum is reported . These cells were able to phagocyte latex beads, iron particles, fluoresceinated zymosan and yeast . Furthermore, non-adherent synoviocytes were capable of being infected by the obligate intracellular parasite of peripheral monocytes Leishmania donovani . Indirect immunofluorescence experiments with specific anti-human monocyte (OKM1) antibody and specific antisynoviocyte serum, showed the presence of common surface structures between synoviocytes A cells and peripheral monocytes . Fifty five percent of the synoviocytes were also positive for HLA Dr antiserum . Analysis by two dimensional gel electrophoresis showed that peripheral monocytes and synoviocytes secreted identical polypeptides in vitro . These results strongly suggest a relationship between synoviocytes A and mononuclear phagocyte system.

J Am Mosq Control Assoc, 1986 Sep, 2(3), 289 - 91
Growth of three mosquitoes on two larval diets measured by protein accumulation; Van Handel E; Larval growth recorded as accumulation of protein was measured in Aedes aegypti, Culex nigripalpus and Culex quinquefasciatus, raised on liver powder or dry brewer's yeast . In the early stages all 3 species grew faster on liver powder, but at pupation there was no significant difference in protein content between diets and among species . Aedes aegypti pupated one day ahead of Culex.

Jpn J Pharmacol, 1986 Sep, 42(1), 35 - 41
Inhibitory effect of N-(3,4-dimethoxycinnamoyl)anthranilic acid on release of SRS from alveolar macrophages in vitro; Yasuoka S et al.; The effects of N-(3,4-dimethoxycinnamoyl)anthranilic acid (N-5') on the release of the slow-reacting substance (SRS) by zymosan- or Ca ionophore-stimulated rat and human alveolar macrophages (AM) were examined in vitro . Disodium cromoglycate (DSCG) was used as a control . N-5' at concentrations of 10(-4)-10(-3) M significantly inhibited the release of SRS from both rat and human AM stimulated by zymosan . N-5' had almost the same inhibitory effect when added to the AM culture system at any time from 180 min before to 30 min after the addition of zymosan . N-5' (10(-4)-10(-3) M) also significantly inhibited the release of SRS by Ca ionophore-stimulated rat AM . N-5' (10(-6)-10(-3) M) had no significant effect on phagocytosis of yeast particles by rat AM . DSCG (10(-6)-10(-3) M) did not inhibit the release of SRS from the zymosan-stimulated rat AM . N-5' was concluded to have a relatively specific inhibitory effect on the non-immunological release of SRS from stimulated AM . It is postulated that N-5' inhibits the process of release of SRS from AM by acting after the initial stage.

Mol Cell Biol, 1986 Aug, 6(8), 2774 - 83
Primary structure of human ribosomal protein S14 and the gene that encodes it; Rhoads DD et al.; Chinese hamster ribosomal protein S14 cDNA was used to recognize homologous human cDNA and genomic clones . Human and Chinese hamster S14 protein sequences deduced from the cDNAs are identical . Two overlapping human genomic S14 DNA clones were isolated from a Charon 28 placental DNA library . A fragment of single-copy DNA derived from an intron region of one clone was mapped to the functional RPS14 locus on human chromosome 5q by using a panel of human X Chinese hamster hybrid cell DNAs . The human S14 gene consists of five exons and four introns spanning 5.9 kilobase pairs of DNA . Polyadenylated S14 transcripts purified from HeLa cell cytoplasma display heterogeneous 5' ends that map within noncoding RPS14 exon 1 . This precludes assignment of a unique 5' boundary of RPS14 transcripts with respect to the cloned human genomic DNA . Apparently HeLa cells either initiate transcription at multiple sites within RPS14 exon 1, or capped 5' oligonucleotides are removed from most S14 mRNAs posttranscription . In contrast to the few murine ribosomal protein and several other mammalian housekeeping genes whose structures are known, human RPS14 contains a TATA sequence (TATACTT) upstream from exon 1 . Three related short sequence motifs, also observed in murine and yeast ribosomal protein genes, occur in this region of the RPS14 gene . RPS14 introns 3 and 4 both contain Alu sequences . Interestingly, the Alu sequence in intron 3 is located slightly downstream from a chromosome 5 deletion breakpoint in one human X hamster hybrid clone analyzed.

Mycopathologia, 1986 Aug, 95(2), 93 - 100
Effects of hyperthermia on phagocytosis and intracellular killing of Sporothrix schenckii by polymorphonuclear leukocytes; Hiruma M et al.; The effects of hyperthermia on phagocytosis and killing of Sporothrix schenckii by polymorphonuclear leukocytes (PMNs) were investigated in order to clarify the mechanism of local thermotherapy for sporotrichosis . Yeast cells of S . schenckii, PMNs and serum were incubated at 37 degrees C or 40 degrees C for 2 or 4 hours . Rate of phagocytosis and killing rate (rate of germination) were estimated, and their processes were observed by transmission electron microscopy . There was no effect of hyperthermia on the phagocytosis rate, but the killing rate increased significantly at 40 degrees C . Electron microscopic examination showed an increase of granularity in the yeast cytoplasm, elongation and fragmentation of the cell membrane . The ultrastructural changes were basically identical under both temperatures, but the degree of these changes was higher at 40 degrees C than at 37 degrees C . Although both intact and degenerated yeasts were found in the same conditions, their transient forms were few, suggesting that the PMN-killing process was completed promptly.

Genetics, 1986 Aug, 113(4), 1009 - 19
Evaluating quantitative variation in the genome of Zea mays; Rivin CJ et al.; Genomic diversity within the species Zea mays has been examined by measuring the variation in the repetitive component of the nuclear genome among North American inbred lines and varieties . This was done by preparing a set of clones of repetitive maize sequences that differ in function, molecular arrangement and multiplicity and then using these as probes for quantitative hybridization to DNA from various maize genotypes . The comparison showed that the majority of repeated sequences are markedly variable in copy number among the ten maize strains tested.The clone sample contained the rDNA and 5S genes, the major repeat of the chromosome knobs, sequences functioning as origins of DNA replication in yeast (ARS sequences) and randomly cloned sequences of unknown function and chromosomal location . The sequences ranged in reiteration frequency from 200 to greater than 10(5) copies and included both tandemly arrayed and dispersed repeats . The copy numbers were measured by hybridizing labeled cloned sequences to aliquots of high molecular weight genomic DNA that were applied to nitrocellulose filters through a slotted template (slot blotting) . The hybridization signal on an autoradiogram occurred in a narrow band that could be scored reliably with a densitometer . This provided a rapid method of determining the abundance of particular repeated sequences in individual plants and plant populations . Using this technique, we found that the copy number of repeated sequences of all types generally varied among the strains by two- to threefold, although at least one sequence showed no detectable variation . In contrast to the variability found between strains, individuals within an inbred line or variety were found to be indistinguishable in terms of specific sequence multiplicity . Each genotype has a different pattern of copy numbers for the set of repeated sequence clones, and this pattern is characteristic of all individuals of a particular genotype . The data also show that the copy number of each sequence varies independently . No strains had uniformly high or low copy numbers for the entire set of probes.

Am J Physiol, 1986 Aug, 251(2 Pt 1), G195 - 200
Muscarinic receptor size on smooth muscle cells and membranes; Collins SM et al.; The loss of {3H}quinuclidinyl benzilate ({3H}QNB) binding following high-energy radiation was used to compare the muscarinic receptor size on single smooth muscle cells isolated by collagenase digestion from the canine stomach and on plasma membranes derived from intact gastric smooth muscle without exposure to exogenous proteolysis . Radiation inactivation of galactose oxidase (68 kdaltons), yeast alcohol dehydrogenase (160 kdaltons), and pyruvate kinase (224 kdaltons) activities were used as molecular-weight standards . Radiation inactivation of {3H}QNB binding to rat brain membranes, which gave a target size of 86 kdaltons, served as an additional control . In isolated smooth muscle cells, the calculated size of the muscarinic receptor was 80 +/- 8 kdaltons . In contrast, in a smooth muscle enriched plasma membrane preparation, muscarinic receptor size was significantly smaller at 45 +/- 3 kdaltons . Larger molecular sizes were obtained either in the presence of protease inhibitors (62 +/- 4 kdaltons) or by using a crude membrane preparation of gastric smooth muscle 86 +/- 7 kdaltons).

Nippon Yakurigaku Zasshi, 1986 Aug, 88(2), 77 - 84
{Pharmacological studies on proglumetacin maleate, a new non-steroidal anti-inflammatory drug . (2) . Analgesic and antipyretic effects}; Ono N et al.; Analgesic and antipyretic effects of proglumetacin maleate (PGM), a new indomethacin (IND) derivative, were investigated in comparison with those of IND on an equimolar-dose basis . The suppression of phenylquinone-induced writhing in mice by PGM was about 0.8 and 2 times as potent as that by IND when given 1 and 4 hr before the phenylquinone injection, respectively . The analgesic activity of PGM in rat silver nitrate arthritis was about 1.5 times more potent than that of IND . PGM was slightly less active in rat adjuvant arthritic pain than IND . On the other hand, PGM provoked a dose-dependent antipyretic effect on the yeast-induced fever in rats within the dose range without affecting the normal body temperature . Furthermore, PGM showed a significant antipyretic effect on LPS-febrile rabbits . Generally, the antipyretic effect of PGM was moderate as compared with that of IND . These analgesic and antipyretic actions of orally administered PGM may be mainly due to its active metabolite, IND . The above results indicate that PGM may be useful for inflammatory diseases associated with pain and/or fever.

Environ Res, 1986 Aug, 40(2), 274 - 84
The effects of ozone inhalation on the immunological response of selenium- and vitamin E-deprived rats; Eskew ML et al.; Deficiencies in vitamin E (E) or Se result in immune alterations, possibly due to reduction of antioxidant activity . Such reductions might greatly compromise the ability of the immune system to deal with additional oxidant stress, as encountered during exposure to air pollutants such as ozone (O3) . To study possible interactions of these oxidative stresses on immune function, male Long-Evans hooded rats were maintained 5 weeks on torula yeast-based diets, with or without the addition of E or Se . Each dietary group was subdivided into O3-exposed and nonexposed groups . Two different regimens of O3 exposure were used: continuous (1.0 ppm, 8 hr/day for 7 days) or intermittent (2.0 ppm, 8 hr/day for 4 days, 2-4 days in ambient air followed by 1 day of exposure prior to sacrifice) . Exposure to O3 in either regimen resulted in increased numbers of cells recovered by pulmonary lavage . With continuous exposure this increase was due to macrophage influx and, with intermittent exposure, due to influx of both macrophages and neutrophils . Combined deficiency of E and Se led to an enhanced ability of spleen and lung cells to mediate antibody-dependent cell-mediated cytotoxicity (ADCMC) . In animals deficient in E, but not Se, O3 exposure depressed spleen cell ADCMC . Deficiencies of either E or Se also depressed lymphocyte response to mitogens . Although intermittent exposure to O3 caused no changes in mitogen response, in animals exposed continuously to O3 there was a significant enhancement of this response.

J Med Vet Mycol, 1986 Aug, 24(4), 297 - 304
A diffusion chamber technique for testing of antifungal drugs against Sporothrix schenckii in vivo; Schaude M et al.; Growth of the yeast form of Sporothrix schenckii (ATCC 14804) was determined in diffusion chambers with 0.45 and 3.0 micron pore size over a period of 24 to 192 h after subcutaneous implantation into mice . Numbers of S . schenckii in 0.45 micron chambers increased significantly by 192 h when inocula of 10(3) and 10(5) colony forming units were implanted . In chambers with a pore size of 3.0 microns, only a slight decrease of fungal growth occurred, although host cells readily passed the filter membrane and phagocytosed yeast-form cells . The activities of amphotericin B, ketoconazole, itraconazole, ICI 153.066, vibunazole and potassium iodide against S . schenckii in implanted chambers were determined in terms of their effects on S . schenckii . ICI 153.066, ketoconazole, itraconazole and amphotericin B significantly reduced the numbers of reisolated S . schenckii in both types of chambers . There was a slight activity with vibunazole but none with potassium iodide.

Proc Natl Acad Sci U S A, 1986 Aug, 83(16), 5871 - 4
Role for topoisomerases in the release of DNA into the detergent-soluble fraction of eukaryotic cells; Zhang LH et al.; Detergent-soluble DNA is the fraction (2-4%) of DNA that is released into the supernate upon mild detergent lysis . It is nonmitochondrial in origin . It labels efficiently with deoxy{3H}ribonucleosides and the labeling is prevented by inhibitors of polymerase alpha and ribonucleotide reductase . In previous publications we have characterized detergent-soluble DNA from splenocytes of immunologically activated mice . In this publication we show that incorporation of {3H}thymidine into detergent-soluble DNA is prevented by pretreatment with novobiocin, 4'-(9-acridinylamino)methanesulfon-m-anisidide (m-AMSA), and teniposide (VM26), three inhibitors of type II topoisomerases . Camptothecin, an inhibitor of type I topoisomerases, also reduces incorporation of {3H}thymidine but only to 50% of control levels . In addition to affecting incorporation of {3H}thymidine, preincubation with the topoisomerase II inhibitors m-AMSA and VM26 alters the amount of DNA recovered in the detergent-soluble fraction . At low concentrations of m-AMSA the amount of detergent-soluble DNA increases somewhat, whereas at higher drug concentrations a marked decrease is observed . Treatment with VM26 results in diminished amounts of DNA being released into the detergent-soluble fraction as well . However, maximal inhibition of detergent-soluble DNA release by VM26 requires the presence of camptothecin . Therefore, we suggest that topoisomerases play an important role in making a small part of lymphocyte chromatin detergent labile . Furthermore, these results are consistent with recent studies demonstrating a role for topoisomerases in yeast replication . Thus, the newly synthesized portion of detergent-soluble DNA may arise as DNA replication intermediates not yet stabilized into mature chromatin.

J Cell Biol, 1986 Aug, 103(2), 621 - 30
Isolation and partial characterization of a 110-kD dimer actin-binding protein; Ueno T et al.; Two Triton-insoluble fractions were isolated from Acanthamoeba castellanii . The major non-membrane proteins in both fractions were actin (30-40%), myosin II (4-9%), myosin I (1-5%), and a 55-kD polypeptide (10%) . The 55-kD polypeptide did not react with antibodies against tubulins from turkey brain, paramecium, or yeast . All of these proteins were much more concentrated in the Triton-insoluble fractions than in the whole homogenate or soluble supernatant . The 55-kD polypeptide was extracted with 0.3 M NaCl, fractionated by ammonium sulfate, and purified to near homogeneity by DEAE-cellulose and hydroxyapatite chromatography . The purified protein had a molecular mass of 110 kD and appeared to be a homodimer by isoelectric focusing . The 110-kD dimer bound to F-actin with a maximal binding stoichiometry of 0.5 mol/mol of actin (1 mol of 55-kD subunit/mol of actin) . Although the 110-kD protein enhanced the sedimentation of F-actin, it did not affect the low shear viscosity of F-actin solutions nor was bundling of F-actin observed by electron microscopy . The 110-kD dimer protein inhibited the actin-activated Mg2+-ATPase activities of Acanthamoeba myosin I and myosin II in a concentration-dependent manner . By indirect immunofluorescence, the 110-kD protein was found to be localized in the peripheral cytoplasm near the plasma membrane which is also enriched in F-actin filaments and myosin I.

Eur J Biochem, 1986 Aug 1, 158(3), 547 - 53
The resolution of heterogeneous fluorescence of multitryptophan-containing proteins studied by a fluorescence-quenching method; Stryjewski W et al.; From acrylamide quenching results, analyzed by an itterative non-linear least-squares method, we have shown that the fluorescence of multitryptophan-containing proteins, such as horse-liver alcohol dehydrogenase, 3-phosphoglycerate kinase and lysozyme, can be resolved for different segmental contributions, each characterized by collisional (Ki) and static (Vi) quenching constants . The ability to resolve the heterogeneous fluorescence of proteins makes it possible to follow changes in dynamics of the individual residues . In yeast 3-phosphoglycerate kinase, which contains only two tryptophan residues, three fluorescent fractions, characterized by different accessibility to the quencher, were observed . Two of them are assigned to one of the tryptophan residue . This may be interpreted in terms of conformational fluctuations, which facilitate the access of acrylamide molecules to the buried tryptophan residues.

Mutat Res, 1986 Aug-Sep, 171(2-3), 177 - 83
Differential staining of chromosomes and spindle cannot be used as an assay to determine the effect of cancer promoters on primary cultures of human fibroblasts; Kirsch-Volders M; The differential staining technique allows simultaneous visualization of chromosomes and spindle fibers . The relative sensitivity of this test for chemicals with high tumor-promoting activity (TPA and iodoacetic acid) or for low tumor-promoting chemicals (4 beta-PDD and alpha-phorbol) was analyzed . Moreover, to ensure a complementary reference system, metals with possible mutagenic (As greater than Hg greater than Cd) and carcinogenic (As greater than Cd = Hg) properties were tested in parallel . As and Hg, which are known as spindle poisons, do induce, as expected, a disaggregation of the spindle fibers; with Cd, no clear-cut disappearance of the spindle is observed . As far as the cancer promoters are concerned, there is no evidence that their ability to induce mitotic aneuploidy in yeast would be confirmed by the induction of spindle disappearance and/or abnormal chromosome segregation in primary cultures of human fibroblasts.

Anticancer Res, 1986 Jul-Aug, 6(4), 785 - 90
Effects of dietary selenium compounds on benzo (a)-pyrene-induced forestomach tumours and whole-blood glutathione peroxidase activities in C3H mice; Bergman K et al.; Selenium (Se) compounds have shown an inhibitory effect on chemically induced tumours in several laboratory models and there is an inverse epidemiological relationship between Se status and certain types of cancer . Little is known about the influence of Se on the development of stomach cancer . Three different forms of dietary Se, selenomethionine, sodium selenite, and high-selenium yeast were investigated as possible inhibitors of benzo(a)pyrene-induced forestomach tumours in mice . The effects of sodium selenite in combination with vitamin E, and of Se-deficiency were also studied . None of the dietary modifications had any effect on tumour incidence or number . Marked elevations of whole-blood glutathione peroxidase (GSH-Px) activities were observed in animals supplemented with all Se-compounds . High-selenium yeast caused the largest increase of GSH-Px activity followed by sodium selenite and selenomethionine . The results indicate that the inhibitory effect of Se on carcinogenesis may be specific with respect to organ site or tumour cell examined.

Proc Natl Acad Sci U S A, 1986 Jul, 83(14), 5277 - 81
Experimental metastasis in nude mice of NIH 3T3 cells containing various ras genes; Bradley MO et al.; These studies have compared the ability of NIH 3T3 cells containing different ras oncogenes to form tumor nodules in the lungs of nude mice after tail vein injection . The genes studied include the normal cellular and bladder tumor ras genes, recombinant viral/cellular ras genes, recombinant yeast/mammalian ras genes, and a constructed gene with yeast RAS1 sequences significantly modified by deletions and an oncogenic mutation . The results show that NIH 3T3 cells containing these genes readily form lethal tumor nodules in the lungs of nude mice after tail vein injection . No control NIH 3T3 cells formed lung tumors within 66 days . Although there were some quantitative differences in the potencies of the various lines, the striking conclusion is that NIH 3T3 cells transformed by either normal or activated mammalian ras genes form approximately equal numbers of experimental lung metastases . In addition, cells transformed by a significantly modified yeast RAS1 gene containing a purposefully introduced oncogenic mutation were also equally active in this assay . The amount of p21 (the 21-kDa protein encoded by ras), as measured by immunoprecipitation, was approximately the same in the parent lines before injection as in the tumors recovered after injection . This result indicates that there is no selection for metastatic sublines containing larger quantities of p21 . Transfection of EJ bladder tumor ras DNA into NIH 3T3 cells followed by injection 3 days later into the tail veins of nude/beige mice indicated that the EJ ras gene can confer a metastatic phenotype within 3.5 cell generations without selection or clonal growth in vitro . Thus, the biochemical changes initiated after introduction of the c-Ha-ras gene into NIH 3T3 cells result in the almost immediate acquisition of phenotypes necessary for experimental metastasis.

Acta Cytol, 1986 Jul-Aug, 30(4), 425 - 9
Phialophora verrucosa-induced subcutaneous phaeohyphomycosis . Fine needle aspiration findings; Schnadig VJ et al.; A 34-year-old woman on immunosuppressive therapy presented with a subcutaneous, cystic lesion on the dorsum of the right foot . Cytologic examination of material obtained by fine needle aspiration (FNA) revealed a mixture of acute and granulomatous inflammation as well as brown-pigmented fungi in the form of budding yeast, pseudohyphae and septate hyphae . The findings suggested subcutaneous phaeohyphomycosis (phaeomycotic cyst) . Culture grew Phialophora verrucosa . The cytologic, histologic and cultural findings are given . This case demonstrates that phaeohyphomycosis can be diagnosed by FNA but that fungal culture is necessary to establish the identity of the etiologic agent . This appears to be the first case of P . verrucosa-induced subcutaneous phaeohyphomycosis reported in the Western Hemisphere.

Proc Natl Acad Sci U S A, 1986 Jul, 83(14), 5155 - 9
A measure of the similarity of sets of sequences not requiring sequence alignment; Blaisdell BE; Determination of first- and second-order Markov chain homogeneity of sets of nuclear eukaryotic DNA sequences, both coding and noncoding, finds similarities imperceptible to the standard Needleman-Wunsch base matching or dot-matrix algorithms . These measures of the similarities of the distributions of adjacent pairs or triplets are in agreement with accepted evolutionary-tree topologies . Hierarchical clustering of the distributions of doublets of 30 miscellaneous coding sequences gives clusters in reasonable agreement with accepted biological classifications . In addition to similarity by homology, there is also observed similarity of disparate genes in the same organism--for example, all three disparate yeast genes (two enzymes and actin) form a well-distinguished cluster.

J Infect, 1986 Jul, 13 Suppl A, 15 - 8
Efficacy and safety of recombinant hepatitis B vaccine in infants born to HBsAg-positive mothers; Yeoh EK et al.; Infants born to HBsAg-positive women were randomly assigned to receive either the plasma-derived hepatitis B vaccine or the yeast-derived recombinant hepatitis B vaccine within 12 h of birth, simultaneously with HBIg . The second and third doses of both vaccines were given at 1 and 6 months of age respectively . Two hundred and twenty-two infants received two doses, and 80 infants received three doses of either vaccine . Five infants were found to be HBsAg-positive at 6 months of age . Four of these infants had received the plasma vaccine and the remaining infant the recombinant vaccine . Anti-HBs was present in all the HBsAg-negative infants at 6 months of age . The titres of anti-HBs were similar in both groups of infants who had received either the plasma or the recombinant hepatitis B vaccine . No major adverse reactions were reported with either vaccine . The preliminary results from this study suggest that the yeast-derived recombinant hepatitis B vaccine is safe, immunogenic and at least as effective as the plasma-derived hepatitis B vaccine in preventing the HBsAg carrier state in infants born to HBsAg-positive women.

Toxicol Appl Pharmacol, 1986 Jul, 84(3), 533 - 40
Pathways for the bioactivation of aliphatic nitriles to free cyanide in mice; Kaplita PV et al.; Reports from several laboratories agree that many, but not all, aliphatic nitriles undergo hepatic biotransformation in mice and rats to release free cyanide, but the mechanisms at work in these reactions remain in doubt . We have used primarily n-butyronitrile, propionitrile, and their respective alpha-carbon-hydroxylated homologs, propionaldehyde cyanohydrin and lactonitrile, to examine this question in mice . Pretreatment of mice with the hepatic microsomal enzyme inducers, pregnenolone-16 alpha-carbonitrile, troleandomycin, and isosafrole, or with the cytochrome P-450-depleting agent, cobaltous chloride, did not influence the mortality of mice given single doses of nitriles . Repeated injections of aspirin or sodium salicylate in water failed to protect mice against death by the nitriles . Dimethyl sulfoxide, however, was effective in reducing mortality after nitrile administration . Repeated injections of 4-methylpyrazole or disulfiram protected mice against death after nitriles . Most of the treatment regimens successful against the nitriles also protected against death due to the cyanohydrins . The cyanohydrins were more acutely toxic than their parent nitriles, produced death much more rapidly, and resulted in the same toxic signs, suggesting that they are intermediates in the bioactivation pathway leading to free cyanide . The cyanohydrins appeared to serve as weak substrates for yeast alcohol dehydrogenase, however, incubation of them with either yeast or horse liver alcohol dehydrogenase did not increase the rate of cyanide release over that in incubations where the enzymes were absent . The slow rate of cyanide release due to spontaneous hydrolysis interfered with the determinations of alcohol dehydrogenase activity, but it cannot account for the rapid action and high toxicity of the cyanohydrins in vivo, or for the efficacy of the treatment regimens which protected against death . It appears unlikely that prostaglandin synthetase or alcohol dehydrogenase are importantly involved in nitrile bioactivation . The same active process, however, appears to be responsible both for alpha-carbon hydroxylation and for the subsequent degradation of the resulting cyanohydrins to release free cyanide . It is far more efficient in mediating the latter reaction than the former.

Ann Hum Genet, 1986 Jul, 50 ( Pt 3), 197 - 206
Immunological cross-reactivity of alcohol dehydrogenase (ADH) isozymes with rabbit immune sera against horse and human ADH subunits; Adinolfi A et al.; Rabbit immune sera raised against denatured forms of horse liver alcohol dehydrogenase and of human ADH5 isozyme were found to react with the denatured subunits of all the human ADH isozymes regardless of their class . The immune serum against the human ADH isozyme cross-reacted also with horse ADH subunits and, at appropriate dilutions, both the immune sera reacted with denatured yeast ADH, suggesting that common structures have been preserved in these molecules over a long evolutionary period . The immune sera partially reacted also with the respective antigens in their native conformation, indicating that some 'sequential' epitopes are expressed on the surface of the folded proteins.

Eur J Biochem, 1986 Jul 1, 158(1), 57 - 62
Structure and expression of ubiquitin genes in higher plants; Gausing K et al.; cDNA clones encoding ubiquitin were isolated from a barley leaf cDNA library using a mammalian ubiquitin cDNA clone as probe . The nucleotide sequence of one of the clones codes for 2.2 perfect repeats of the 76-amino-acid-long ubiquitin protein with an extra lysine residue at the C-terminus . The barley ubiquitin amino acid sequence differs from the animal sequence at three positions and from the yeast sequence at two positions . The ubiquitin poly(precursor) are coded by a multigene family with 8-10 genes that produce four or five different size messengers between 700 and 2000 nucleotides in length . The large poly(A)-rich RNAs are constitutively expressed in vegetative tissues whereas the 700-nucleotide messenger(s) were only detected in tissues containing dividing cells.

Biochim Biophys Acta, 1986 Jun 23, 871(3), 285 - 92
Proton and nitrogen-15 NMR studies of ferricytochrome c cyanide complexes: remarkable conservation of the heme environment among organisms of diverse origin; Behere DV et al.; The native ferric and cyanide-bound ferric forms of nine vertebrate and two yeast cytochromes c have been investigated by high-resolution proton nuclear magnetic resonance spectroscopy . Spectral comparisons have been made among the cytochromes with emphasis on the signal positions for heme and amino acid ligand protons . Consistent with earlier more limited studies of native ferric cytochromes c, the paramagnetically shifted proton NMR signals show little variation among species with up to 50% substitution of amino acids . Proton NMR spectra for the cyanide complexes also show little variation among species . The nitrogen-15 signal for the coordinated cyanide ion is known to be highly variable among other hemoproteins, but the signal covers a range of only 855 to 865 ppm (nitrate ion reference) for vertebrate cytochromes c and 884 to 886 ppm for yeast cytochromes c . The cyanide ligand probe thus reports an amazing conservation of the heme and proximal ligand environment among the cytochromes . Comparative proton and nitrogen-15 chemical shift values are consistent with a slightly stronger proximal histidine imidazole hydrogen bond to an amino acid carbonyl function than is the case for hemoglobin and myoglobin.

J Biol Chem, 1986 Jun 15, 261(17), 8063 - 9
Multiple forms and cellular localization of Drosophila DNA topoisomerase II; Heller RA et al.; Purified type II topoisomerase from Drosophila melanogaster embryos was reported earlier to contain a major polypeptide of 166,000 daltons and several smaller peptides between 132,000 and 145,000 daltons (Shelton, E . R., Osheroff, N . and Brutlag, D . L . (1983) J . Biol . Chem . 258, 9530-9535) . Using purified topoisomerase II we have raised antibodies against the 132,000-166,000-dalton cluster of polypeptides . In this paper we demonstrate that at least three of these polypeptides are also present in embryos immediately upon lysis . Using antigen-affinity purified antibody from the cluster of purified topoisomerase II antigens, we have also discovered several smaller polypeptides in the molecular size range of 30,000-40,000 daltons in embryo extracts . These observations suggest the presence of multiple forms of DNA topoisomerases in the cell . In addition, we demonstrate that purified Drosophila topoisomerase II antibody recognizes yeast topoisomerase II antigens expressed by lambda gt 11-yeast topoisomerase II recombinants (Goto, T . and Wang, J . C . (1984) Cell 36, 1073-1080) establishing a structural homology between yeast and Drosophila enzymes . Antibody preparations were also used to localize the distribution of topoisomerase II in polytene nuclei . In contrast with the distribution of topoisomerase I which is located primarily at puffs, the Drosophila topoisomerase II is distributed generally along the chromosomes paralleling the distribution of DNA itself.

J Biol Chem, 1986 Jun 5, 261(16), 7479 - 84
Chromosomal protein HMG-17 . Complete human cDNA sequence and evidence for a multigene family; Landsman D et al.; Antibodies elicited against chromosomal protein HMG-17, purified from calf, were used to screen a human lambda gt11 cDNA expression library and isolate the full length cDNA coding for this protein . Sequence analysis reveals that the nucleotide distribution along this cDNA is highly asymmetric . The amino acid sequence, deduced from the reading frame, reveals that the human HMG-17 is, respectively, 96 and 92% homologous with the calf and chicken protein . The amino acid substitution are conservative suggesting evolutionary constraints on the conformation of the protein . The human genome contains 35-50 HMG-17 gene copies which, as revealed by Southern analysis, are distributed at several loci . Northern analysis of total RNA isolated from 3 human cell lines, indicates that each cell contains a single-size mRNA coding for this protein . Nucleotide sequences which cross-hybridize, under stringent conditions, with the human HMG-17 cDNA are present in the genome of rodents and absent from the genomes of sea urchin, Drosophila, and yeast . The availability of a probe for the HMG-17 gene may help elucidate the cellular role of this protein which may confer specific conformations to transcribable regions in the genome.

J Med Vet Mycol, 1986 Jun, 24(3), 231 - 3
Red-pigmented Histoplasma capsulatum--an unusual variant; Morris PR et al.; An unusual variant of Histoplasma capsulatum was isolated from a canebrake . The mycelial form produced a red pigment that diffused into the medium and which was also present in the cell walls of the mycelium, microconidia and macroconidia . The yeast cells were not pigmented nor did they produce any pigment.

Yeast, 1986 Jun, 2(2), 87 - 92
A new type of methylamine oxidase: the sole oxidase produced during growth of Sporobolomyces albo-rubescens on primary alkylamines; Sherlock LA et al.; Under conditions known to separate methylamine oxidase from benzylamine oxidase in other yeast strains, only a single oxidase could be detected in Sporobolomyces albo-rubescens . This occurred irrespective of whether methylamine or n-butylamine was the nitrogen source for growth . The oxidase did not attack benzylamine . It was concluded that this organism can only produce a methylamine oxidase . The enzyme was purified to 90% homogeneity and found to have properties significantly different from the methylamine oxidases previously characterised . It lost only 40% of its activity in 30 min at 45 degrees C, whereas methylamine oxidases previously described had half-lives of from 2 to 9 min at 45 degrees C . It showed also a lower activity with short chain 1-aminoalkanes and a higher activity with longer chain 1-aminoalkanes than other methylamine oxidases, and had a significantly smaller subunit molecular weight (57,000 compared with 80,000).

Proc Natl Acad Sci U S A, 1986 Jun, 83(12), 4119 - 23
Characterization of acetylated and acetolyzed glycoprotein high-mannose core oligosaccharides by fast-atom-bombardment mass spectrometry; Tsai PK et al.; Fast-atom-bombardment mass spectrometry has been applied to acetylated neutral and phosphorylated oligosaccharides from yeast glycoproteins and to their acetolysis products . Although acetylation increases the sample molecular weight and the complexity of the spectra, it also enhances the sensitivity of detection, is applicable to samples that contain salt, and is especially useful for analysis of phosphorylated derivatives . Acetylation by trifluoroacetic anhydride/glacial acetic acid is particularly convenient and can be done rapidly on a small amount of material . Acetolysis by acetic anhydride/glacial acetic acid/H2SO4 is done on the acetylated oligosaccharides, and the acetylated fragments are recovered by solvent extraction and immediately subjected to mass spectrometry . The methodology allows molecular weight determinations and sequence analysis by acetolysis to be carried out on a few micrograms of isolated oligosaccharide in a few hours.

J Natl Cancer Inst, 1986 Jun, 76(6), 1157 - 62
Interactions of selenium deficiency, vitamin E, polyunsaturated fat, and saturated fat on azoxymethane-induced colon carcinogenesis in male F344 rats; Reddy BS et al.; The effect of the interaction of selenium deficiency, excess vitamin E, and type of fat on colon carcinogenesis induced by azoxymethane (AOM) was studied in male F344 rats . The experimental diets, based on a Torula yeast diet and containing 20% stripped corn oil or 20% stripped lard, were as follows: 1) selenium deficient with adequate (50 mg/kg diet) vitamin E, 2) selenium deficient with excess (750 mg/kg diet) vitamin E, 3) selenium adequate with adequate vitamin E, and 4) selenium adequate with excess vitamin E . Starting at about 3 weeks of age, animals were fed the experimental diets, and at 7 weeks of age all animals except the vehicle-treated controls were given sc injections of AOM (15 mg/kg body wt) once weekly for 2 weeks . Animals were fed the experimental diets until termination of the experiment . Selenium deficiency significantly inhibited the incidence (percentage of animals with tumors) and multiplicity (tumors per animal) of colon adenocarcinomas and adenomas, whereas excess vitamin E had no effect on colon carcinogenesis . There was no interaction between the selenium status and vitamin E; the selenium status and type of fat; vitamin E and type of fat; and among selenium status, vitamin E, and type of fat.

Horm Metab Res, 1986 Jun, 18(6), 415 - 7
A possible competition between 5'-monodeiodination of thyroxine and the respiratory burst in human granulocytes; Nagy JT et al.; Thyroxine (T4) pretreatment of A 23187-stimulated human granulocytes in 10(-5)-10(-6) M concentration range inhibited the superoxide anion production of these cells . T4 increased the level of oxidized form of glutathione, whereas the intracellular level of the reduced form decreased . A similar alteration in the ratio of the oxidized to reduced forms of glutathione was detected in granulocytes during yeast cell phagocytosis . In addition, conversion of T4 to triiodothyronine (T3) was also inhibited during phagocytosis . A possible competition between 5'-monodeiodination of T4 and the oxidative burst of human granulocytes is discussed.

J Leukoc Biol, 1986 Jun, 39(6), 679 - 86
Tetrahydrocannabinol-induced suppression of macrophage spreading and phagocytic activity in vitro; Lopez-Cepero M et al.; The effects of tetrahydrocannabinol (THC) on several parameters of macrophage function in vitro were assessed . Delta 9 THC added to cultures of normal mouse peritoneal cells in vitro affected the ability of the cells to spread on glass surfaces and also had some effect on their ability to phagocytize yeast . These effects were dose related . A concentration of 20 micrograms of THC almost completely inhibited macrophage spreading, but it also decreased viability and the total number of these cells . Doses of 10 or 5 micrograms of THC also inhibited spreading but had little effect on cell viability or number . A dose of 1.0 microgram of THC had some inhibitory effect on spreading and the lowest dose affecting spreading appeared to be about 0.05 micrograms per culture . Higher doses of THC were necessary to inhibit phagocytosis of yeast particles as determined by direct microscopic examination or use of radiolabeled yeast as the test particles . These results indicate that several readily measured functions of macrophages may be suppressed by THC.

Arch Int Physiol Biochim, 1986 Jun, 94(2), 161 - 72
Anomeric specificity of mammalian hexokinase; Giroix MH et al.; The anomeric specificity of hexokinase was examined in crude homogenates of rat parotid gland, erythrocytes and pancreatic islets . At 8 degrees C, the alpha/beta ratio in maximal velocity averaged 0.73, 0.66 and 0.75 in the parotid, erythrocytes and pancreatic islets, respectively . Hexokinase displayed a greater affinity for alpha- than beta-D-glucose as judged from three criteria: the Km value, the reaction velocity measured with mixtures of the two anomers and their effect upon the phosphorylation of D-{U-14C} glucose in anomeric equilibrium . The latter procedure yielded an alpha/beta ratio in Km close to 0.51, 0.49 and 0.39 in parotid, erythrocytes and pancreatic islets, respectively . Within the limits of this study, the anomeric specificity of mammalian hexokinase would appear to be a mirror image of that of yeast hexokinase.

Mycopathologia, 1986 Jun, 94(3), 157 - 61
Immunocytochemical staining of Histoplasma capsulatum at the electron microscopic level; Patino MM et al.; Rabbits were immunized with histoplasmin emulsified in Freund's complete adjuvant . Antibody raised in these rabbits was exposed to Histoplasma capsulatum yeast cells, either in tissue culture medium, or after in vitro or in vivo phagocytosis by mouse macrophages . The sites of antibody binding were identified using an immunoperoxidase technique . At least two sites of antibody binding were identified, one to the fungal cell wall and the other to the outer cell membrane . Within 6 h after phagocytosis by macrophages, fungal cell walls appeared roughened, with what appeared to be cell wall antigen released into the phagolysosome, appearing associated with the phagolysosome membrane, and possibly adjacent macrophage cytoplasm . Similar staining of fungal antigen was noted in alveolar macrophages which had ingested Histoplasma capsulatum after a respiratory challenge . This method may be useful in detailing the host/pathogen interactions which occur in human pulmonary histoplasmosis.

Nucleic Acids Res, 1986 May 27, 14(10), 4077 - 94
A cDNA clone of the hnRNP C proteins and its homology with the single-stranded DNA binding protein UP2; Lahiri DK et al.; A cDNA clone which expresses a protein that cross-reacts immunologically with the human C1 and C2 hnRNP core proteins has been isolated . The clone was selected by a sensitive immunochemical assay employing an avidin-biotin complex for detection, and identified as a clone for the hnRNP C proteins by a highly sensitive antibody select assay that is described here . The clone contains 677 nucleotides, and, as shown by northern blotting, is derived from a 1.5 Kb poly(A)+ mRNA . There are regions of strong homology between the human and mouse genes, weak homology is seen with chicken DNA, and very little, if any, homology can be detected with Drosophila, Artemia, sea urchin, or yeast DNAs . Two peptides (a total of 24 amino acids) of the calf thymus single-stranded DNA binding protein UP2 show perfect homology with the deduced amino acid sequence of the clone, suggesting that UP2 is related to the hnRNP C proteins . There is also a region that has a sequence very similar to two regions of the single-stranded DNA binding protein UP1 that contain proposed DNA binding sites.

J Biol Chem, 1986 May 25, 261(15), 6705 - 11
Structural studies on the eukaryotic chain initiation factor 2 from rabbit reticulocytes and brine shrimp Artemia embryos . Phosphorylation by the heme-controlled repressor and casein kinase II; Mehta HB et al.; In contrast to reticulocyte polypeptide chain initiation factor 2 (eIF-2), the Artemia factor retains activity in the presence of Mg2+ or after phosphorylation of its alpha-subunit by rabbit reticulocyte heme-controlled repressor (Mehta, H . B., Woodley, C . L., and Wahba, A . J . (1983) J . Biol . Chem . 258, 3438-3441) . Furthermore, we have so far been unable to demonstrate a requirement for a GDP/GTP nucleotide exchange factor with Artemia eIF-2 . In order to explain these differences we compared the structure of eIF-2 from Artemia and rabbit reticulocytes by using one- and two-dimensional phosphopeptide and iodopeptide maps . Partial trypsin digestion of the alpha-subunit of Artemia eIF-2 after phosphorylation by the heme-controlled repressor generates a 4000 Mr phosphopeptide . Upon extensive trypsin digestion, the two-dimensional phosphopeptide maps of the alpha-subunits for the reticulocyte and Artemia factors are indistinguishable, whereas the iodopeptide maps are different . In addition, immunoblotting indicates that there is no consistent cross-reactivity of the reticulocyte subunits with antibodies prepared in rabbits against the Artemia eIF-2 subunits . A casein kinase II activity was isolated from Artemia embryos that phosphorylates the beta-subunit of reticulocyte eIF-2, but specifically phosphorylates the alpha-subunit of eIF-2 preparations from several non-mammalian sources, including Artemia, yeast, and wheat germ embryos . Since this kinase phosphorylates a site distinct from that recognized by the heme-controlled repressor, and this phosphorylation does not alter the ability of Artemia eIF-2 to undergo nucleotide exchange, caution must be exercised when interpreting the significance of eIF-2(alpha) phosphorylation in non-mammalian cells.

Eur J Biochem, 1986 May 15, 157(1), 77 - 81
Nucleotide sequence of the ARGRII regulatory gene and amino acid sequence homologies between ARGRII PPRI and GAL4 regulatory proteins; Messenguy F et al.; We report here the DNA sequence of the ARGRII gene, one of the three regulatory genes involved in controlling the anabolism and catabolism of arginine in yeast . This gene encodes a protein of 880 amino acids with a deduced molecular mass of about 100 kDa . The ARGRII protein shows significant homology with two other regulatory proteins of yeast, PPRI and GAL4.

J Immunol, 1986 May 15, 136(10), 3706 - 9
Development and characterization of antiserum to murine granulocyte-macrophage colony-stimulating factor; Mochizuki DY et al.; The expression in yeast of a cDNA clone encoding murine granulocyte-macrophage colony-stimulating factor (GM-CSF) has made possible the purification of large quantities of this recombinant protein . Rabbits immunized with pure recombinant GM-CSF generated antibodies that were shown to be specific for both recombinant GM-CSF and GM-CSF isolated from natural sources . Other lymphokines, including IL 1 beta, IL 2, IL 3, and recombinant human GM-CSF did not react with the antiserum . The antiserum, together with recombinant GM-CSF that had been radiolabeled with 125I to high specific activity, formed the foundation for a rapid, sensitive, and quantitative radioimmunoassay specific for murine GM-CSF . Furthermore, the antiserum was found to inhibit the biologic activities of GM-CSF as measured in both a bone marrow proliferation assay and a colony assay, and thus should prove to be a useful reagent for dissecting the complex growth factor activities involved in murine hematopoiesis.

J Biol Chem, 1986 May 15, 261(14), 6255 - 9
Phosphorylation and degradation of exogenous phosphatidylinositol incorporated into Friend erythroleukemic cells; Hohengasser CJ et al.; Phosphatidyl{2-3H}inositol (PtdIns) obtained from rat skeletal muscle and yeast was introduced into Friend erythroleukemic cells by use of the PtdInstransfer protein or by spontaneous route . The mammalian PtdIns incorporated by the transfer protein appeared metabolically inert while the spontaneously incorporated PtdIns was both phosphorylated to PtdIns-4-phosphate (i.e . 30% of the total PtdIns incorporated) and converted into lyso-PtdIns (i.e . 20% of the total PtdIns incorporated); formation of PtdIns, 4,5-bisphosphate was minimal . The extensive metabolism of the spontaneously incorporated PtdIns strongly suggests that this PtdIns does not rapidly equilibrate with the endogenous PtdIns pools . A similar spontaneous incorporation of yeast PtdIns was accompanied by a negligible degree of phosphorylation and hydrolysis . Evidence is provided that this difference in metabolism reflects the absence of arachidonate in the yeast PtdIns.

Biochem Biophys Res Commun, 1986 May 14, 136(3), 1148 - 54
N-alkanes induce the synthesis of cytochrome P-450 mRNA in Candida maltosa; Wiedmann B et al.; In the yeast Candida maltosa the level of cytochrome P-450 was 100-300-fold higher in alkane-grown cells than in glucose-grown ones (detected by a radioimmunoassay) . It was shown immunochemically (1) that this was not the result of an assembly of preexisting apoenzyme with the prosthetic heme group . By cell-free translation of total poly(A)RNA in a wheat germ system and subsequent immunoprecipitation it was shown that the amount of mRNA coding for cytochrome P-450 paralleled its concentration in the cell.

Biochim Biophys Acta, 1986 May 5, 866(4), 179 - 203
How proteins get into microbodies (peroxisomes, glyoxysomes, glycosomes); Borst P; All microbody proteins studies, including one microbody membrane protein, are made on free polysomes and imported post-translationally . This holds for animal tissues, plants, and fungi . The majority of microbody protein sub-units are synthesized in a form not detectably different from mature sub-units . In five cases a larger precursor protein has been found . The position of the extra piece in this precursor is not known . In two of the five cases, processing of the precursor is not coupled to import; in the other three this remains to be determined . It is not even known whether information in the prepiece contributes to topogenesis, or serves other purposes . Microbody preparations from Neurospora, plant tissue and rat liver can take up some newly synthesized microbody proteins in vitro . In most cases uptake is inefficient . No special requirements for uptake have been established and whether a receptor is involved is not yet known . Several examples have been reported of peroxisomal enzymes with a counterpart in another cell compartment . With the exception of catalase, no direct evidence is available in any of these cases for two isoenzymes specified by the same gene . In the Zellweger syndrome, a lethal hereditary disease of man, characterized by a lack of peroxisomes, the levels of several enzymes of lipid metabolism are strongly decreased . In contrast, D-amino-acid oxidase, L-alpha-hydroxyacid oxidase and catalase levels are normal . The catalase resides in the cytosol . Since there is no separate gene for cytosolic catalase, the normal catalase levels in Zellweger cells show that some peroxisomal enzymes can mature and survive stably in the cytosol . It is possible that maturation of the peroxisomal enzyme in the cytoplasm can account for the finding of cytosolic catalase in some normal mammalian cells . The glycosomes of trypanosomes are microbodies that contain a glycolytic system . Comparison of the glycosomal phosphoglycerate kinase with its cytosolic counterpart has shown that these isoenzymes are 93% homologous in amino-acid sequence, but less than 50% homologous to the corresponding enzymes of yeast and mammals . This implies that few alterations are required to direct a protein into microbodies . This interpretation is supported by the evidence for homology between some microbody and mitochondrial isoenzymes in other organisms mentioned under point 4 . The major changes of the glycosomal phosphoglycerate kinase relative to the cytosolic enzyme are a large increase in positive charge and a C-terminal extension of 20 amino acids.(ABSTRACT TRUNCATED AT 400 WORDS)

Int J Pept Protein Res, 1986 May, 27(5), 443 - 8
A "helix-scissors" mechanism for the hinge-bending conformational change in phosphoglycerate kinase; Blake CC et al.; X-ray studies of phosphoglycerate kinase (EC 2.7.2.3, PGK) have shown that the enzyme's single polypeptide chain is organized into two separate domains that correspond to the N- and C-terminal halves of the chain . Substrate binding studies and the incorporation of the complete amino acid sequence of horse-muscle PGK into its X-ray model suggest that the C-domain is an ADP/ATP binding unit and that the N-terminal domain contains the phosphoglycerate binding site and the active site located in a prominent cluster of positively charged residues . Because the distance between these two sites is 12-15 A, a hinge-bending of 10 degrees--20 degrees has been proposed to bring the two sites together for catalysis . Independent solution studies of yeast PGK have shown that the radius of gyration decreases significantly on the formation of the ternary complex . This change has been interpreted in terms of a 9 degrees--12 degrees rotation about a hinge in the interdomain region that brings the two domains together . We suggest here a structural basis for the proposed hinge-bending that involves the rotation of the two helices that form the domain interface about their contact normal carrying their respective domains with them.

Clin Allergy, 1986 May, 16(3), 251 - 7
Clinical and immunological characteristics of pre-school asthma; Loftus BG et al.; Ninety-two children, sixty-two boys and thirty girls, aged 18 months to 6 years with frequent asthma were studied . Nine were born prematurely and two were ventilated in the neonatal period; two had been admitted to hospital with acute bronchiolitis . Most had an early onset of symptoms, forty-two developing asthma in infancy . Although overall growth was normal, 63% had some chest deformity . Half had past or present eczema and just over half had chronic rhinitis . Upper respiratory infection appeared to be an important precipitant of attacks in all, and 88% had exercise-induced asthma . The majority were atopic, 74% had elevated serum IgE levels, skin-prick tests were positive in 61 of 77 tested and half had eosinophilia . However, allergic factors were clearly identified as causing asthma attacks in only 24% . Immunoglobulin deficiencies were found in thirteen and defective yeast opsonization in eighteen; these children showed no other obvious differences from the remainder . Eighty-two children were followed up and after 1 year fifty-one were receiving inhaled steroids . Measurement of serum immunoglobulin, eosinophil count and skin-prick testing had little influence on management.

Cell Biol Int Rep, 1986 May, 10(5), 383 - 7
Possible sequences for nuclear accumulation of parvoviral proteins; Lederman M et al.; Parvoviral genomes have been searched for sequences which may code for the nuclear transport of viral proteins . Sequences similar to those which regulate the nuclear transport of T antigen and yeast mating type protein were detected within the sequences coding for capsid and non-capsid proteins.

Nippon Yakurigaku Zasshi, 1986 May, 87(5), 527 - 36
{The analgesic and antipyretic effects of a non-steroidal antiinflammatory agent, EB-382, in experimental animals}; Fujiyoshi T et al.; The analgesic and antipyretic effects of EB-382 as a new non-steroidal antiinflammatory agent were examined in mice and rats . EB-382 had an equipotent inhibition to ibuprofen on the writhing syndrome caused by acetic acid, phenylquinone and acetylcholine in mice, but phenylbutazone was less potent in these experiments . EB-382 had a much more potent inhibition on the pain by the Randall-Selitto method and silver nitrate-induced arthritic pain in rats than ibuprofen and phenylbutazone . EB-382 had no analgesic effect on the pain of non-treated foot by the Randall-Selitto method in rats and by the hot-plate method in mice . EB-382 had a much more potent inhibition on the yeast-induced chronic inflammatory and adjuvant arthritic pains in rats than ibuprofen and phenylbutazone . The antipyretic activity of EB-382 was almost equipotent to that of ibuprofen in rats . EB-382 had no effect on the normal body temperature in rats, which was different from aminopyrine . The above results suggest that EB-382 will be a useful analgesic agent with an antipyretic antiinflammatory activity in clinical studies.

J Exp Med, 1986 May 1, 163(5), 1085 - 99
Murine eosinophil differentiation factor . An eosinophil-specific colony-stimulating factor with activity for human cells; Lopez AF et al.; A purified murine lymphokine, eosinophil differentiation factor (EDF), was found to be a selective stimulus for the clonal proliferation and differentiation of murine eosinophil progenitor cells, establishing it as the murine eosinophil colony-stimulating factor (Eo-CSF) . EDF was also active on human eosinophil progenitors and mature blood eosinophils, but had no effect on neutrophil or macrophage precursor cells, nor on blood neutrophils . In culture of human bone marrow cells, EDF stimulated equal numbers and equal sizes of eosinophil colonies to develop when compared with human placental conditioned medium, a source of human CSFs, suggesting that all responsive progenitor cells were stimulated . Clone transfer experiments and the linear relationship between number of bone marrow cells plated and colonies produced confirmed that the action of EDF was directly on eosinophil progenitor cells . EDF increased the capacity of human blood eosinophils, but not neutrophils, to kill antibody-coated tumor cells and to phagocytose serum-opsonized yeast cells . This functional activation was associated with the enhanced expression of functional antigens (GFA-1, GFA-2, and the receptor for C3bi) on eosinophils . The possession by EDF (Eo-CSF) of all the properties expected of a human eosinophil CSF raises the possibility that a human analog of this molecule exists, and is involved in the regulation of production and function of human eosinophils in vivo.

Proc Natl Acad Sci U S A, 1986 May, 83(10), 3121 - 5
A mouse tumor-specific transplantation antigen is a heat shock-related protein; Ullrich SJ et al.; A tumor-specific transplantation antigen has been purified to homogeneity from the cytosol of a methylcholanthrene-induced tumor, Meth A . The purified antigen is highly immunogenic and specific against challenge with Meth A, providing greater than 95% inhibition of tumor growth in immunized syngeneic mice . Immunofluorescence analysis of Meth A showed that the antigen is a highly abundant cytosolic protein but that it is also present at the cell surface and, therefore, accessible to the host's immune system . The antigen consists of two polypeptide isoforms present in equimolar amounts, having similar masses (84 and 86 kDa), pI values (4.95 and 4.90), and amino acid compositions . Both are phosphoproteins, and neither is glycosylated . The NH2-terminal sequences of the two isoforms are identical except that each chain contains a portion of unique sequence . Comparison of the NH2-terminal and CNBr-fragment sequence data to the sequences of the yeast and Drosophila heat shock proteins (Hsp90 and Hsp83, respectively) reveals that 73 of 91 residues compared are identical . In addition, an anti-Meth A tumor antigen serum that defects the isoforms from a variety of tumors also immunoprecipitates proteins of identical mass and pI from both normal and heat-shocked mouse embryo cells.

Med Hypotheses, 1986 May, 20(1), 37 - 50
The nature of the scrapie agent; Adams DH; There now seems little doubt that the infective agent of scrapie cannot be accommodated within current concepts of virology/molecular biology . It is proposed: that the basic infective entity is a nucleic acid fragment (oligonucleotide) of some 40 bp coupled with specific (but host encoded) protein totalling approximately 10(5) daltons, a significant proportion of which is in the form of proteolipid; that the nucleic acid fragment reprograms the host cell on the chemically switched microprocessor network analogy already proposed; that the nucleic acid fragment has no initiation sequence for replication: it is therefore non-infective; that the nucleic acid fragment can replicate when associated with the specific protein component because the resulting complex is able to displace mobile genetic element flanking sequences (similar to the yeast delta sequence) . The function of the protein is to provide a scaffolding which allows the nucleic acid fragment to be assimilated into the replication cycle of the mobile genetic element as a whole.

Mutat Res, 1986 May, 165(3), 191 - 8
A mammalian cell variant in which 3-aminobenzamide does not potentiate the cytotoxicity of dimethyl sulphate; Murray B et al.; Variants of mouse leukaemia L1210 cells have been isolated in which cytotoxicity to dimethyl sulphate is not fully potentiated by ADP-ribosyl transferase inhibitor 3-aminobenzamide, as occurs in normal L1210 cells . These variants were selected after mutagenesis by growing the cells in dimethyl sulphate and 3-aminobenzamide . The characterisation of one of these variants is described . Variant 3 cells repair low doses of DNA damage in the presence of ADP-ribosyl transferase inhibitors . The Vmax of the ADP-ribosyl transferase enzyme in these cells is only increased 35% compared to normal wild-type L1210 cells . The basal DNA ligase I activity is increased 66% above wild-type whereas DNA ligase II activity appears to be unchanged . The most striking observation, however, is that the DNA ligase II activity is not increased after dimethyl sulphate treatment as occurs in wild-type L1210 cells . It seems that by increasing DNA ligase I levels these cells can survive DNA damage in the presence of 3-aminobenzamide . This variant (mutant) provides genetic evidence for our previously published hypothesis that (ADP-ribose)n biosynthesis is required for efficient DNA repair after DNA damage by monofunctional alkylating agents, because ADP-ribosyl transferase activity regulates DNA ligase activity . This variant is the first mammalian cell reported in which DNA ligase activity is altered, as far as we are aware . In yeast, a DNA ligase mutant has a cell division cycle (cdc) phenotype . Presumably, DNA ligase is essential for DNA synthesis, repair and recombination . The present variant provides further evidence that in mammalian cells, DNA ligase II activity is related to ADP-ribosyl transferase activity.

Science, 1986 Apr 25, 232(4749), 506 - 8
Induction of macrophage tumoricidal activity by granulocyte-macrophage colony-stimulating factor; Grabstein KH et al.; Monocytes are a subpopulation of peripheral blood leukocytes, which when appropriately activated by the regulatory hormones of the immune system, are capable of becoming macrophages--potent effector cells for immune response to tumors and parasites . A complementary DNA for the T lymphocyte-derived lymphokine, granulocyte-macrophage colony-stimulating factor (GM-CSF), has been cloned, and recombinant GM-CSF protein has been expressed in yeast and purified to homogeneity . This purified human recombinant GM-CSF stimulated peripheral blood monocytes in vitro to become cytotoxic for the malignant melanoma cell line A375 . Another T cell-derived lymphokine, gamma-interferon (IFN-gamma), also stimulated peripheral blood monocytes to become tumoricidal against this malignant cell line . When IFN-gamma activates monocytes to become tumoricidal, additional stimulation by exogenously added lipopolysaccharide is required . No such exogenous signals were required for the activation of monocytes by GM-CSF.

J Biol Chem, 1986 Apr 25, 261(12), 5396 - 403
On the recognition of helical RNA by cobra venom V1 nuclease; Lowman HB et al.; The V1 nuclease from cobra venom preferentially hydrolyzes double helical RNA and has been used extensively for detecting RNA secondary structure . To increase the utility of this enzyme as an RNA structure probe, we have investigated its properties and substrate specificity, using assays for polynucleotide hydrolysis based on fluorescent polynucleotide derivatives . Enzymatic activity requires both Na+ and Mg2+, with optima at 100 and 0.3 mM, respectively . From the sharp decrease in enzyme activity above 100 mM Na+ we estimate that 3-4 ionic interactions between the protein and polynucleotide phosphates take place . Analysis of products remaining after extensive V1 digestion also shows that the minimum size substrate is 4-6 nucleotides long . Helical RNAs and DNAs have Michaelis constants a factor of 3-10 times lower than most single-stranded RNAs . However, poly(epsilon A) has a Michaelis constant equal to the best synthetic double helices tested and is hydrolyzed at a rate comparable to helical RNA . The major V1 cutting sites in yeast tRNAPhe have Michaelis constants lower than any synthetic polymers . These data suggest that V1 nuclease recognizes any 4-6-nucleotide segment of polynucleotide backbone with an approximately helical conformation, but does not require that the bases be paired in a helix . A few single-stranded V1 cleavage sites are known in tRNA and rRNA, and their structures are consistent with the suggested V1 recognition site.

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