Yeasts

Mol Cell Biol, 1984 Dec, 4(12), 2686 - 96
The two beta-tubulin genes of Chlamydomonas reinhardtii code for identical proteins; Youngblom J et al.; The two beta-tubulin genes of the unicellular green alga Chlamydomonas reinhardtii are expressed coordinately after deflagellation and produce two transcripts of 2.1 and 2.0 kilobases . Full-length cDNA clones corresponding to the transcript of each gene were isolated . DNA sequences were obtained from the cDNA clones and from cloned tubulin gene fragments . Both genes contained 1,332 base pairs of coding sequence, with only 19 nucleotide differences between the genes . Because all the differences occurred at the third base position of a codon and did not change the predicted amino acid sequence, we concluded that both beta-tubulin genes code for the same protein of 443 amino acids . The predicted amino acid sequence is 89 and 72% homologous with beta-tubulins from chicken and yeast cells, respectively . Each gene had three intervening sequences, which occurred at identical positions . Although the first two intervening sequences were not conserved between the two genes, the nucleotide sequence of the third intervening sequence was 89% conserved between the genes . The codon usage in the tubulin genes of C . reinhardtii was very biased: only 37 different codons were used . Striking differences occurred between the codons used in these nuclear genes and C . reinhardtii chloroplast genes.

EMBO J, 1984 Dec 1, 3(12), 2779 - 86
Termination of the ovalbumin gene transcription; LeMeur MA et al.; RNA transcribed in vivo from the chicken ovalbumin gene has been analyzed in detail in the 3' region of the gene using nuclease S1 mapping and Northern blotting . We describe two new additional minor polyadenylation sites leading to pre-messenger RNA with lengths of approximately 7900 and 9700 nucleotides . Hybridization of RNA transcribed in vitro from oviduct nuclei with various immobilized DNA fragments covering the 3' region of the gene indicates that greater than 90% of transcription terminates in a discrete region of 170 bp located approximately 900 bp downstream from the 3' end of the last exon . Two copies of a sequence homologous to the one proposed for yeast transcription termination are present within the region where transcription of the ovalbumin gene terminates.

Proc Natl Acad Sci U S A, 1984 Dec, 81(23), 7412 - 6
Molecular cloning and nucleotide sequence of cDNA for rat ornithine carbamoyltransferase precursor; Takiguchi M et al.; Messenger RNA of rat ornithine carbamoyltransferase (EC 2.1.3.3), a mitochondrial matrix enzyme, was enriched by immunoprecipitation of rat liver free polysomes, and recombinant plasmids were prepared from the enriched mRNA by a vector-primer method . The cDNA clones for ornithine carbamoyltransferase were identified by hybrid-arrested translation and hybrid-selected translation . One of the clones, designated pOTC-1, contained a 1.6-kilobase insert and hybridized to a mRNA of approximately equal to 1.8 kilobases in rat liver . The cDNA clone was subjected to nucleotide sequence analysis . The deduced amino acid sequence indicates that the ornithine carbamoyltransferase precursor consists of the mature enzyme of 322 amino acid residues and an NH2-terminal peptide extension (presequence) of 32 amino acid residues . The presequence contains 8 basic amino acid residues, no acidic residues, and no hydrophobic amino acid stretch . The amino acid sequence of the rat ornithine carbamoyltransferase was compared with the recently reported sequence of the human enzyme {Horwich, A . L., Fenton, W . A., Williams, K . R., Kalousek, F., Kraus, J . P., Doolittle, R . F., Konigsberg, W . & Rosenberg, L . E . (1984) Science 224, 1068-1074} . The sequences of the mature enzyme portion are 93% identical, whereas those of the presequences are 69% identical . There are two highly conserved segments in the presequences of the rat and human enzymes . One of the two conserved segments is significantly similar to a segment of the presequence of yeast mitochondrial elongation factor EF-Tu . These results suggest that the homologous segments are important for the proteins that are synthesized in the cytosol to be transported into the mitochondrial matrix.

Z Naturforsch {C}, 1984 Nov-Dec, 39(11-12), 1042 - 7
Preparation of an affinity chromatographic system for the separation of ADP binding proteins; Bieber E et al.; {4-(3-Bromoacetylpyridinio)-butyl}adenosine pyrophosphate as a structural analog of NAD+ reacts covalently with the sulfhydryl groups of thiopropyl agarose . 10-20 mumol can be bound to 1 ml gel . Stabilization of the insoluble coenzyme is attained by treatment with sodium boro hydride (NaBH4) . This complex when applied to column chromatography, allows the separation of various dehydrogenases as a result of their different complex stability coefficients . Alcohol dehydrogenase from liver, lactate dehydrogenase, and adenylate kinase, which all bind to the ADP-analog residues of the gel matrix, can thus be separated by different salt gradients . Alcohol dehydrogenase from yeast, however, does not form a complex and can easily be eluted from the column with phosphate buffer . Glyceraldehyde-3 phosphate and aldehyde dehydrogenases can be eluted by the addition of NAD+ or NADH to the buffer . The uncharged 1,4-dihydropyridine ring of the reduced coenzyme produces a more stable complex with the dehydrogenases than the oxidized form.

Res Vet Sci, 1984 Nov, 37(3), 353 - 4
Diversity in nicotinamide-adenine dinucleotide requirement for the growth of different strains of Mycoplasma synoviae; Yagihashi T et al.; The type strain WVU 1853 and field strains SG, N26 and A642 of Mycoplasma synoviae were examined for their requirement for nicotinamide-adenine dinucleotide (NAD) for in vitro growth . All the strains grew and could be repeatedly passaged in Frey broth medium supplemented with filter-sterilised NAD . In modified Frey broth medium from which NAD was omitted and broth medium for which the supplements including yeast extract and NAD were autoclaved, only strains N26 and A642 could be grown and passaged . The growth curves of strain N26 determined in broth media with and without NAD were similar . These results indicated that there are differences in NAD-requirement for in vitro growth among strains of M synoviae.

Poult Sci, 1984 Nov, 63(11), 2217 - 24
Effect of dietary composition and estradiol implants on hepatic microsomal mixed function oxidase and lipid deposition in growing chicks; Takahashi K et al.; Three experiments were conducted to investigate the effect of diet and estradiol (E2) administration on hepatic microsomal mixed function oxidase (MFO) activity, E2 metabolism, and liver lipid content in male broiler chicks . Broiler chicks (3 weeks of age) were fed either a corn-soybean (CS) diet or a diet containing fish meal, alfalfa meal, and torula yeast (FAY) for 19 days in Experiments 1 and 3 and for 14 days in Experiment 2, respectively . Half of the chicks were implanted with tubes containing E2 . In all experiments when the chicks were estrogenized, feeding FAY significantly lowered liver lipid content and plasma E2 concentration . Activity of hepatic microsomal aniline hydroxylase and content of cytochrome P-450 were significantly increased by feeding FAY with or without E2 administration . The chicks fed the CS diet had a significantly lower content of cytochrome P-450 when E2 was administered . Activities of aminopyrine demethylase and nicotinamide adenine dinucleotide phosphate, reduced (NADPH)-cytochrome C reductase did not differ significantly between the diets . In in vitro studies, conversion of 14C-E2 into the water soluble fraction was significantly increased in microsomes from chicks fed the FAY diet as compared to ones from chicks fed the CS diet . The results suggest that some of the hepatic microsomal functions on the CS diet are modified by the change in diet composition and that these modifications are probably associated with E2 metabolism and occurrence of fatty liver.

J Exp Med, 1984 Nov 1, 160(5), 1261 - 72
Trichomonas vaginalis is dependent on uptake and degradation of human low density lipoproteins; Peterson KM et al.; Human plasma low density lipoprotein uptake by the urogenital pathogen, Trichomonas vaginalis, was examined . Rapid binding and internalization of 125I-labeled low density lipoproteins by live T . vaginalis was observed at 37 degrees C . Data showing parasite degradation of the internalized apoproteins and lipid accumulation following low density lipoprotein uptake was obtained . Acquisition of low density lipoproteins was by a trichomonad surface protein that possessed a molecular weight of greater than 250,000 . The receptor is specific for apolipoprotein CIII, a component of high, low, and very low density lipoprotein subfractions . Low density lipoproteins in a semi-defined medium of trypticase, nucleic acid precursors, vitamins, and maltose promoted T . vaginalis growth and multiplication at rates and levels equal to the yeast extract-trypticase-serum complex medium routinely used for culture of trichomonads . HeLa cell membranes as a source of lipids were unable to sustain T . vaginalis organisms . These data demonstrate host lipoprotein internalization by T . vaginalis via a specific uptake mechanism.

Eur J Immunol, 1984 Nov, 14(11), 997 - 1002
Cell surface events which may initiate lysosomal enzyme secretion by human monocytes; Leoni P et al.; Freshly isolated and subsequently matured human monocytes secreted lysosomal hexosaminidase in response to exposure to IgG-Sepharose, but not certain derivatized control Sepharoses . The cells bound selectively to the surface of IgG-Sepharose (and not the control Sepharoses) but because of the large size of the particles, could not ingest them . Since the soluble IgG was covalently linked to the Sepharose and free soluble IgG was not an inducer of secretion, the secretion was thus induced directly at the cell surface . Zymosan, a yeast cell wall particle which contains a mannan, was also able to induce secretion in the various monocyte stages under study . It could even bind to the cell surface of fresh monocytes which lacked the receptor for mannose-terminated glycoproteins, and induce secretion in these cells . The mannose receptor appeared as monocytes matured, and the Ir number on the surface was increased by the action of lymphokines . Although zymosan-induced secretion could be inhibited by mannose and certain other sugars, these seemed to have some complex metabolic effects in human monocytes (which previous work with mouse macrophages has not revealed) . Thus, it was not possible to demonstrate whether zymosan could initiate secretion directly by interaction at the monocyte surface mannose glycoprotein receptor.

Arch Microbiol, 1984 Nov, 140(1), 83 - 5
Mode of action of glyphosate in Candida maltosa; Bode R et al.; The broad-spectrum herbicide glyphosate inhibits the growth of Candida maltosa and causes the accumulation of shikimic acid and shikimate-3-phosphate . Glyphosate is a potent inhibitor of three enzymes of aromatic amino acid biosynthesis in this yeast . In relation to tyrosine-sensitive 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase and dehydroquinate synthase, the inhibitory effect appears at concentrations in the mM range, but 5-enolpyruvylshikimate 3-phosphate (EPSP) synthase is inhibited by micromolar concentrations of glyphosate . Inhibition of partially purified EPSP synthase reaction by glyphosate is competitive with respect to phosphoenolpyruvate (PEP) with a Ki-value of 12 microM . The app . Km for PEP is about 5-fold higher and was 62 microM . Furthermore, the presence of glyphosate leads to derepression of many amino acid biosynthetic enzymes.

Mol Cell Biol, 1984 Nov, 4(11), 2289 - 97
Role of intron-contained sequences in formation of moloney murine leukemia virus env mRNA; Hwang LS et al.; Formation of the Moloney murine leukemia virus envelope mRNA involves the removal of a 5,185-base pair-long intron . Deletion analysis of two Moloney murine leukemia virus-derived expression vectors revealed the existence of two short regions within the viral intron which are required for the efficient formation of the spliced RNA species . One region was present upstream from the 3' splice junction, extended at least 85 nucleotides beyond the splice site, and was not more than 165 nucleotides long . As yeast polymerase II introns, the Moloney murine leukemia virus intron contains the sequence 5'-TACTAAC-3' 15 nucleotides upstream from the 3' splice site . A second region located in the middle of the intron, within a 560-nucleotide-long sequence, was also essential for formation of the spliced RNA species . The efficient splicing of the env mRNA in the absence of expression of viral genes raises the possibility that similar mechanisms are used to remove introns of (some) cellular genes.

Br J Dermatol, 1984 Nov, 111(5), 603 - 7
The response of seborrhoeic dermatitis to ketoconazole; Ford GP et al.; A randomized double-blind placebo-controlled cross-over study was made of ketoconazole 200 mg daily in nineteen patients with seborrhoeic dermatitis . All had scalp lesions and sixteen had seborrhoeic dermatitis at other sites . Responses were measured by clinician and patients independently, using a linear analogue scale . Body and scalp lesions and itch regressed considerably and significantly with ketoconazole in all but five patients, three of whom subsequently responded to a higher dose . The patients studied with seborrhoeic dermatitis had been sent by their family doctors in answer to a request for patients with dandruff, and the clinical difference between the two was found to be only of degree . Three patients with dandruff without erythema were studied separately using the same study design: all three responded similarly to those with seborrhoeic eczema . It is concluded that Pityrosporum yeast infection is the immediate cause of seborrhoeic dermatitis and that dandruff is its mildest manifestation.

Cell, 1984 Nov, 39(1), 141 - 8
Developmental regulation of a Dictyostelium gene encoding a protein homologous to mammalian ras protein; Reymond CD et al.; We have cloned, sequenced, and examined the regulation of a Dictyostelium gene encoding a protein homologous to mammalian ras proteins . The Dictyostelium, yeast, and mammalian proteins have homologous N-terminal regions and less conserved C-terminal regions . We have used DNA probes and a polyclonal antibody to examine the differential accumulation of ras RNA and protein through development . The gene encodes two mRNAs (0.9 and 1.2 kb) that are differentially expressed . The 1.2 kb RNA is found in vegetative cells and disappears rapidly upon initiation of development . Later, both RNAs accumulate preferentially in prestalk cells . The level of the Dd-ras protein remains constant until early culmination and then decreases . Like other prestalk genes, Dd-ras can be induced with cAMP in the absence of cell contact . When aggregated cells are dissociated, both mRNAs decrease . Upon addition of cAMP, the 1.2 kb mRNA reaccumulates at a higher level than that in normal developing cells . The presence of the Dd-ras protein in vegetative cells corroborates other reports suggesting a possible function during cell growth . The sustained level of Dd-ras protein in prestalk cells suggests an additional role during differentiation.

Am J Med, 1984 Oct 30, 77(4D), 39 - 43
Esophageal, gastric, and intestinal candidiasis; Trier JS et al.; Gastrointestinal Candida infection is more prevalent than previously recognized . It is most often seen in patients with underlying impairment of the immune system but may also occur in apparently normal individuals . Esophageal involvement is most common, presenting with odynophagia, dysphagia, or bleeding . Gastric Candida infection may cause diffuse mucosal involvement or focal invasion of benign gastric ulcers . Intestinal candidiasis is uncommon and poorly characterized . The diagnosis is usually established by visualizing the characteristic yeast or mycelial forms in endoscopic brushings and biopsies . Oral nystatin is effective therapy in many patients, but other antifungal agents may be needed in extensive or persistent disease, especially in immunocompromised patients.

J Biol Chem, 1984 Oct 25, 259(20), 12514 - 8
3-O-methylation of mannose residues . A novel reaction in the processing of N-linked oligosaccharides occurring in Mucor rouxii; Lederkremer GZ et al.; Yeast- and mycelial-form cells of the dimorphic fungus Mucor rouxii incubated with {U-14C}glucose were found to synthesize Man-P-dolichol, Glc-P-dolichol, and Glc3Man9GlcNAc2-P-P-dolichol . The structure of the oligosaccharide moiety of the latter was similar to that of the same compound isolated from other eucaryotic cells . Oligosaccharides that migrated on paper chromatography as Man6-30GlcNAc standards were obtained upon treatment of delipidated proteins with a protease and endo-beta-N-acetylglucosaminidase H . The oligosaccharides that migrated apparently as single substances on paper chromatography could be separated into three different populations by paper electrophoresis in sodium borate buffer . The fastest migrating substances contained only mannose and N-acetylglucosamine residues, whereas the other two contained, in addition, different proportions of 3-O-methylmannose units . The oligosaccharides with the highest content of 3-O-methylmannose residues appeared to be completely resistant to alpha-mannosidase degradation; they were, however, cleaved by endo-beta-N-acetylglucosaminidase H . Mycelial cells synthesized a much higher proportion of 3-O-methylmannose-containing oligosaccharides than yeast cells . Cells incubated with {methyl-14C}methionine were found to label only the N-linked oligosaccharides containing 3-O-methylmannose residues . It is concluded that transfer of Glc3Man9GlcNAc2 to protein is followed by excision of glucose and probably one or two mannose residues, followed by further mannosylation and in some cases also methylation of oligosaccharides . This represents a novel reaction in the processing of N-linked oligosaccharides.

Mech Ageing Dev, 1984 Oct 15, 27(2), 153 - 60
Growth rate and life span in Drosophila . III . Effect of body size and developmental temperature on the biphasic relationship between growth rate and life span; Economos AC et al.; The previous finding of a biphasic relationship between life span and growth rate of Drosophila, developed at 25 degrees C, was confirmed at other temperatures in the usual range (19-28 degrees C) and for development in either a standard medium or one deprived of nutrients but with varying amounts of yeast added on the medium . The role of body size in this relationship was studied by developing flies in a nutrient-less medium with a constant, submaximal amount of added yeast and varying temperature . It was found that under these conditions, which abolished the usual inverse relationship between body size and developmental temperature, body size variations did not account for the observed variations in life span . Thus, corroborating previous studies from this laboratory, body size was ruled out as a causative factor in the life span-growth rate relationship.

Mech Ageing Dev, 1984 Oct 15, 27(2), 143 - 51
Growth rate and life span in Drosophila . II . A biphasic relationship between growth rate and life span; Economos AC et al.; The relationship between growth rate and life span was studied in Drosophila by varying the amount of yeast available to each developing larva at constant temperature, 25 degrees C . With one approach the larvae developed in a standard medium at constant larval density and a varying amount of yeast added on the medium . Across the entire growth rate range covered in this way (10-100 micrograms/day, male flies) imaginal life span depended on growth rate in a biphasic way, the relationship having a parabolic form with a maximum at about 55 to 60 micrograms/day . Similar covariation of growth rate and life span was obtained by varying larval density at a constant amount of added yeast . With both these approaches growth rate variation was due to opposite variations of both components of growth rate, i.e . duration of development and body size . However, development in a medium without nutrients but with a varying amount of added yeast at constant larval density led to a similar biphasic relationship between growth rate and life span although duration of development did not vary . Therefore, the present results are not compatible with the hypothesis that there is a single causal negative relationship between growth rate and life span and demonstrate that duration of development is not a causative factor of the biphasic relationship between growth rate and life span established here.

Mech Ageing Dev, 1984 Oct 15, 27(2), 233 - 7
Age-related changes in cAMP and cGMP levels during phagocytosis in human polymorphonuclear leukocytes; Fulop T et al.; Alterations of cyclic nucleotides have been studied during the incorporation of opsonized yeast cells into human polymorphonuclear leukocytes (PMNL) from both young and aged subjects . In PMNLs obtained from healthy young males the cAMP level rose to a maximal value during the first 15 min and returned to the starting level at the 60th min of phagocytosis . The cGMP level started to rise only at the 30th min of incorporation and enhanced progressively up to the 120th min . In contrast, the elevated cAMP level remained increased during the 120 min of phagocytosis in PMNLs obtained from aged subjects, whereas the cGMP level was not altered during the same period . The basic level of cAMP diminished, while the cGMP level was found to be elevated with ageing in PMNLs . It is concluded that phagocytosis was not impaired with age yet cytotoxic functions were diminished and that the data presented support the idea that cyclic nucleotides may be directly involved only in the latter process.

Eur J Biochem, 1984 Oct 15, 144(2), 387 - 92
Import and processing of cytochrome b-c1 complex subunits in isolated hepatoma ascites cells . Inhibition by Rhodamine 6G; Kolarov J et al.; The import and processing of cytochrome c1 and the iron sulfur protein of the cytochrome b-c1 complex were studied in Zajdela hepatoma ascites cells . Both peptides were synthesized as larger percursor molecules which were approximately 2-3 kDa and 5-6 kDa larger than the mature forms of apocytochrome c1 and apo-iron sulfur protein, respectively . Comparison of these precursors to those reported for functionally homologous peptides in yeast and Neurospora indicate significant size changes have occurred in mammals . Rhodamine 6G, a specific vital stain for mitochondria, is a potent inhibitor of precursor processing in isolated hepatoma cells . Both precursor to cytochrome c1 and precursor to FeS accumulate in the soluble and particulate fractions obtained by digitonin treatment of tumor cells treated with Rhodamine 6G . Appearance of the mature peptides was abolished . The precursors are unstable, however, and disappear from the cytosolic and membrane fractions during a 10 min chase . Comparison of the effects of Rhodamine 6G and carbonylcyanide m-chlorophenylhydrazone on precursor processing shows that: (a) Rhodamine 6G is a more effective inhibitor of processing, (b) it has less of an inhibitory effect on cellular protein synthesis, and (c) it inhibits processing under conditions in which it appears to have little influence on coupled respiration in whole cells . The data suggest that the most likely mode of action of Rhodamine 6G is on the matrix processing step.

Science, 1984 Oct 5, 226(4670), 67 - 70
A gradient of sequence divergence in the human adult alpha-globin duplication units; Hess JF et al.; The nucleotide sequences of the two 5'-homology blocks of human alpha-globin gene duplication units were determined . The sequence difference between the two blocks is essentially zero in the 5' portions, and increases gradually toward the 3' ends until it reaches a value of 18 percent . This gradient of sequence divergence is similar to the distribution of the frequencies of gene conversion along several loci in Ascobolus and yeast . Hot spots for initiation of gene correction processes appear to exist near the 5' ends of the human alpha-globin duplication units . The data provide the physical evidence for polar gene correction process in a mammalian genome.

J Clin Microbiol, 1984 Oct, 20(4), 813 - 4
Taxonomic and nomenclatural evaluation of the genera Candida and Torulopsis; McGinnis MR et al.; It is concluded that the validly published genera Candida and Torulopsis are taxonomically distinct . The recently proposed merger of these two yeast genera is rejected.

Biochem J, 1984 Oct 1, 223(1), 151 - 60
Uptake of mannose-terminated glycoproteins in isolated rat liver cells . Evidence for receptor-mediated endocytosis in hepatocytes; Tolleshaug H et al.; Even though most of the hepatic binding capacity for mannose-terminated glycoproteins has previously been shown to reside in the hepatocytes (not in the non-parenchymal cells), detailed evidence for the specific uptake of mannose-terminated ligands has been lacking . In the present studies, yeast invertase, a large glycoprotein (Mr 270 000) containing about 50% mannose, was shown to be taken up into hepatocytes by receptor-mediated endocytosis . The uptake was saturable and could be specifically inhibited by mannosides or by a Ca2+ chelator . The asialo-glycoprotein receptor was not involved . The low-Mr (13 000) ligand ribonuclease B, which contains a single high-mannose glycan, was not taken up by hepatocytes; however, it was taken up as fast as invertase by non-parenchymal liver cells . After injection of 131I-invertase into a rat in vivo, about one-half of the labelled protein was recovered in the hepatocytes . On a per-cell basis, each endothelial cell contained 3-4 times as much radioactivity as did the hepatocytes . On fractionation of hepatocytes in sucrose gradients, invertase showed a different intracellular distribution from that of asialo-fetuin, in that invertase moved much faster into that region of the gradient where the lysosomes were recovered . This indicates that invertase and asialo-fetuin are not transported intracellularly by identical mechanisms.

Appl Environ Microbiol, 1984 Oct, 48(4), 747 - 50
Isolation of a bioemulsifier from Candida lipolytica; Cirigliano MC et al.; The yeast Candida lipolytica produced an inducible extracellular emulsification activity when it was grown with a number of water-immiscible carbon substrates . Negligible emulsification activity was produced by this yeast when it was grown with glucose as the carbon substrate . In hexadecane-supplemented cultures, emulsification activity was first detected after 36 h of growth, with maximum production after 130 h . A water-soluble emulsification activity was partially purified by repeated solvent extractions of the culture filtrate . This emulsifier, which we named liposan, was primarily composed of carbohydrate . Maximum emulsification activity was obtained when the ratio of hexadecane to liposan was 50:1 . Maximum emulsification activity was obtained from pH 2 to 5 . Liposan was heat stable to temperatures up to 70 degrees C, with a 60% loss in activity after heating for 1 h at 100 degrees C . Liposan effected stable oil-in-water emulsions with a variety of hydrocarbons.

Surgery, 1984 Oct, 96(4), 731 - 7
Percutaneous aspiration and drainage for suspected abdominal infection; Pruett TL et al.; Percutaneous drainage (PCD) of abdominal infection is a therapeutic modality whose role is not well defined . Surgical literature on abdominal infection cites a cumulative mortality rate in the range of 20% to 30%, markedly dissimilar from the 80% to 90% cure rates reported in the literature on PCD . We reviewed the PCD experience at a tertiary teaching hospital from 1981 to 1983 . Fifty-five patients were suspected to have localized abdominal infection and underwent 66 procedures . PCD was attempted after percutaneous needle aspiration produced drainable fluid . Cure is defined by complete resolution of the abdominal process without any surgical intervention . Palliation is defined as acute decompression of the abdominal process permitting an elective corrective procedure to be performed . Failure is defined as false diagnosis, unsuccessful drainage requiring operation, or recurrence of infection . Diagnosis of the abdominal process was successfully made by aspiration in 59/66 (89%) attempts . PCD was curative in 31/66 (47%) attempts and failed or was palliative in 35/66 (53%) . Simple nonfungal, nonfistulous abdominal abscesses were cured with PCD in 25/26 attempts (96%) . PCD failure was encountered in 10 infected organized hematomas or thick phlegmons, nine fungal infections, nine abscesses with enteric communication, and five infected necrotic tumors . Abscesses with an underlying enteric communication were cured in 28%, were palliated in 32% and failed in 32% of PCD attempts . Abscesses with yeast as a major component or with necrotic tumor were never cured with PCD . PCD is a valuable diagnostic and therapeutic tool that is curative in simple abdominal abscesses . Its therapeutic role in complex abdominal infections seems to be limited.

J Cell Biol, 1984 Oct, 99(4 Pt 1), 1535 - 40
Phosphomannosyl receptors may participate in the adhesive interaction between lymphocytes and high endothelial venules; Stoolman LM et al.; Normal and malignant lymphocytes can migrate from the bloodstream into lymph nodes and Peyer's patches . This process helps distribute normal lymphocytes throughout the lymphoid system and may provide a portal of entry for circulating malignant cells . An adhesive interaction between lymphocytes and the endothelium of postcapillary venules is the first step in the migratory process . We have recently shown that the simple sugars L-fucose and D-mannose, and an L-fucose-rich polysaccharide (fucoidin), can inhibit this adhesive interaction in vitro . We now report that mannose-6-phosphate, the structurally related sugar fructose-1-phosphate, and a phosphomannan, core polysaccharide from the yeast Hansenula holstii (PPME) are also potent inhibitors . Inhibitory activity was assessed by incubating freshly prepared suspensions of lymphocytes, containing the various additives, over air-dried, frozen sections of syngeneic lymph nodes at 7-10 degrees C . Sections were then evaluated in the light microscope for the binding of lymphocytes to postcapillary venules . Mannose-6-phosphate and fructose-1-phosphate were potent inhibitors of lymphocyte attachment (one-half maximal inhibition at 2-3 mM) . Mannose-1-phosphate and fructose-6-phosphate had slight inhibitory activity, while glucose-1-phosphate, glucose-6-phosphate, galactose-1-phosphate, and galactose-6-phosphate had no significant activity (at 10 mM) . In addition, the phosphomannan core polysaccharide was a potent inhibitor (one-half maximal inhibition at 10-20 micrograms/ml); dephosphorylation with alkaline phosphatase resulted in loss of its inhibitory activity . Preincubation of the lymphocytes, but not the lymph node frozen sections, with PPME resulted in persistent inhibition of binding . Neither the monosaccharides nor the polysaccharide suppressed protein synthesis nor decreased the viability of the lymphocytes . Furthermore, inhibitory activity did not correlate with an increase in negative charge on the lymphocyte surface (as measured by cellular electrophoresis) . These data suggest that a carbohydrate-binding molecule on the lymphocyte surface, with specificity for mannose-phosphates and structurally related carbohydrates, may be involved in the adhesive interaction mediating lymphocyte recirculation.

Toxicol Appl Pharmacol, 1984 Sep 30, 75(3), 454 - 65
Biochemical, cytological, and histological alterations in rat lung following acute beryllium aerosol exposure; Hart BA et al.; Fischer 344 rats were exposed for 1 hr to an aerosol of BeO generated at a temperature of 560 degrees C . An initial lung burden of 500 +/- 4.1 ng Be was achieved . Animals were killed at 2.5 hr, and 2, 12, and 21 days postexposure . Bronchopulmonary lavage fluids were analyzed biochemically for enzymes, protein, lipids, and sialic acid, and cytologically to determine the composition of the free alveolar cell population . Nonspecific phagocytosis of yeast was measured in adherent macrophages . There were increases in all the biochemical parameters by 2 days postexposure, which peaked by Day 5 and then began to return to control levels . The cytological response on Days 2 and 5 was characterized by polymorphonuclear leucocyte infiltration and a depression in macrophage number and phagocytic activity . By Day 12, increased numbers of newly recruited macrophages with supranormal phagocytic activity populated the lung . During the same period, there was a reduction in lavage protein and lipid levels, perhaps due to a restoration of normal clearance mechanisms . Tissue morphological changes correlated well with the cytological and biochemical alterations.

Biochemistry, 1984 Sep 25, 23(20), 4572 - 80
pH profiles and isotope effects for aconitases from Saccharomycopsis lipolytica, beef heart, and beef liver . alpha-Methyl-cis-aconitate and threo-Ds-alpha-methylisocitrate as substrates; Schloss JV et al.; alpha-Methyl-cis-aconitate (cis-2-butene-1,2,3-tricarboxylate) was converted only to alpha-methylisocitrate (3-hydroxybutane-1,2,3-tricarboxylate) by aconitases from beef liver or S . lipolytica . While the kinetic parameters of beef liver (cytoplasmic) or heart (mitochondrial) aconitases did not vary over the pH range 4.9-9 with the natural substrates, and only slightly with the alpha-methyl substrates, the yeast aconitase exhibited a bell-shaped pH profile with all substrates and for binding of the competitive inhibitor, tricarballylate, with pK values around 7 and 9 . The third pK of the substrates does not affect V/K, showing that these pK's are for catalytic groups on the enzyme . One of these catalytic groups presumably removes a proton to give the carbanion intermediate in the reaction, and the other protonates the hydroxyl group when it is eliminated to give water, possibly with the assistance of the Fe-S center . Beef liver aconitase showed a primary deuterium isotope effect of 1.12 (measured by equilibrium perturbation with deuterated alpha-methylisocitrate) which was pH independent and only slightly greater than the equilibrium isotope effect . Isotope effects with the yeast enzyme were also pH independent but about 1.22 on V/K (or when measured by equilibrium perturbation) and 1.7 on V . These data suggest a kinetic mechanism for beef aconitases in which product release occurs only by displacement by the substrate in a step independent of pH or of the protonation state of the substrate . With the yeast enzyme, product displacement either depends on the protonation state of the catalytic groups on the enzyme or can occur spontaneously at a finite rate.(ABSTRACT TRUNCATED AT 250 WORDS)

J Biol Chem, 1984 Sep 25, 259(18), 11169 - 72
Dicyclohexylcarbodiimide binds to cytochrome b and subunit VIII in soluble complex III from beef heart mitochondria; Clejan L et al.; Incubation of soluble complex III isolated from either yeast or beef heart mitochondria with 25-100 nmol of {14C}dicyclohexylcarbodiimide (DCCD)/nmol of cytochrome b followed by centrifugation through 10% sucrose or precipitation with trichloroacetic acid did not result in any changes in the appearance of the subunits of either complex . The {14C}DCCD was bound to cytochrome b and phospholipids in the yeast complex and with similar kinetics to both cytochrome b and subunit VIII (Mr = 4000-8000) plus phospholipids of the beef complex . Subunit VIII of the beef complex was partially extracted with chloroform:methanol; however, no subunit of this mobility was present in the yeast complex . Incubation of the beef complex in phosphate buffer for short times resulted in a doubling of the {14C}DCCD bound to cytochrome b relative to that to subunit VIII . Preincubation of both complexes with venturicidin prior to treatment with DCCD resulted in a 50% decrease in the binding of {14C}DCCD to cytochrome b . Reisolation of the beef complex III by precipitation with (NH4)2SO4 after incubation with {14C}DCCD resulted in the formation of a new band with an apparent molecular weight of 39,000 even in the zero time control . The {14C}DCCD was bound to subunit VIII and the core proteins but not to cytochrome b at all times, suggesting that precipitation with (NH)2SO4 in the presence of DCCD causes cross-linking of the subunits of complex III.

Biochem Biophys Res Commun, 1984 Sep 17, 123(2), 489 - 96
Uptake of free hemoglobin by rat liver parenchymal cells; Weinstein MB et al.; Parenchymal cells isolated from rat liver are capable of taking up free hemoglobin . Uptake was saturable with a concentration for half-maximal velocity of 1.35 mg/ml (1.99 X 10(-5) M) hemoglobin . At a concentration of 0.088 mg/ml, the endocytic index for hemoglobin uptake was 4.5 microliters/h per mg of cell protein . This may be compared with the rate of fluid pinocytosis by these cells of 0.025 microliter/h per mg of cell protein (determined with yeast invertase as the marker) . Free beta globin chains were also taken up with an endocytic index of 26.7 microliters/h per mg of cell protein at a beta chain concentration of 0.075 mg/ml . Hemoglobin inhibited the uptake of labeled beta globin . Hemoglobin-haptoglobin complex at a concentration of 0.12 mg/ml (as hemoglobin) was cleared at a rate of 0.89 microliter/h per mg cell protein and its uptake was also inhibited by free hemoglobin . We conclude that haptoglobin serves to conserve the iron of hemoglobin by preventing its renal clearance and not by promoting its hepatic uptake.

J Biol Chem, 1984 Sep 10, 259(17), 11153 - 6
Direct photoaffinity labeling of proteins with adenosine 3'-{32P}phosphate 5'-phosphosulfate . Atractyloside inhibits labeling of a Mr = 34,000 protein in an adrenal medullary Golgi fraction; Lee RW et al.; Direct photoaffinity labeling with radioactively labeled adenosine 3'-phosphate 5'-phosphosulfate (PAPS) followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography was used to identify PAPS binding proteins in a Golgi membrane preparation of bovine adrenal medulla . {3'-32P}PAPS was synthesized from adenosine 5'-phosphosulfate (APS) and {gamma-32P}ATP using APS kinase prepared from yeast and was purified by reverse-phase ion pair high performance liquid chromatography . Upon irradiation with UV light, {3'-32P}PAPS, as well as {35S}PAPS under conditions which minimized sulfotransferase-catalyzed incorporation of 35SO4 from {35S}PAPS into proteins, bound selectively to a 34-kDa protein of the Golgi membrane preparation . PAPS binding to the 34-kDa protein was strongly inhibited by the presence of 50 microM atractyloside . The 34-kDa PAPS binding protein therefore appears to be similar to the mitochondrial ATP/ADP translocator with regard to both molecular weight and inhibition by atractyloside of adenine nucleotide binding . Photoaffinity labeling will be useful in the purification and functional identification of the 34-kDa protein.

Mol Biol (Mosk), 1984 Sep-Oct, 18(5), 1380 - 4
{The effect of exogenous tRNA from different sources on the biosynthesis of milk proteins}; Turkovskaia GV et al.; The tRNA dependent cell--free protein--synthesizing system from rabbit differentiated mammary gland has been obtained . The level of protein synthesis including caseins was found to be much higher in the presence of homologous tRNA in comparison with tRNAs from non-differentiated mammary gland, liver, brewer's yeast . The efficiency of translation was shown to depend on the tRNA pool quantitative balance . The addition of tRNA to mammary gland explants causes stimulation of casein synthesis . The level of this stimulation depends on both the origin of tRNA and physiological state of the gland . It is concluded that the functional adaptation of tRNA is a regulatory link in specific protein biosynthesis at the translation level.

Exp Eye Res, 1984 Sep, 39(3), 343 - 54
The purification and properties of human lens glutathione reductase; Latta K et al.; A very simple and rapid method for the purification of human lens glutathione reductase has been developed . The method involves only two steps--affinity chromatography on 2',5'-ADP-Sepharose 4B and gel filtration on Sephacryl S-200 . With whole lenses, the purification achieved is over 18000-fold and 80% of the activity in the tissue homogenate is recovered as an enzyme with a specific activity of 218 IU/mg-1 . Glutathione reductase was purified from the nucleus and cortex of type I cataractous human lenses and the properties of the two enzymes were compared . No differences could be detected between these enzymes in their specific activities, molecular weights, pH-activity profiles, heat labilities, reactivity towards various substrates and kinetic parameters (Vmax and Km) for glutathione, NADPH2 and NADH2 . Therefore, it was concluded that specific alterations in the properties of nuclear glutathione reductase were not responsible for the decreased ability of the lens nucleus to protect itself against oxidative insults . Several different glutathione reductase preparations were examined for their ability to cleave mixed disulphides of lens proteins and glutathione . Only crude tissue extracts (wheat germ and whole lens) were able to cleave the mixed disulphides: no activity was observed with purified lens or yeast glutathione reductases . Therefore, it was concluded that glutathione reductase does not cleave mixed disulphides.

J Clin Microbiol, 1984 Sep, 20(3), 421 - 9
Determination of catalase, peroxidase, and superoxide dismutase within the genus Legionella; Pine L et al.; We examined 40 strains of Legionella for reduced-oxygen scavenging enzymes . Using a simple reaction chamber with a Swinney filter for the Beers and Sizer assay, we determined the catalase activity of live cells grown on buffered charcoal-yeast extract agar . For 29 strains of Legionella pneumophila, the apparent first-order rate constants for catalase ranged from 0.000 to 0.005 . Similarly, low values ranging from 0.001 to 0.005 were observed for Legionella wadsworthii, Legionella oakridgensis, and Legionella gormanii . High catalase activities were found for Legionella jordanis, Legionella longbeachae, Legionella micdadei, and Legionella bozemanii, with first-order rate constant values of 0.010 to 0.035 . Cell-free extracts were analyzed for catalase, peroxidase, and superoxide dismutase . Cell-free extracts of all strains had superoxide dismutase levels ranging from 8.2 to 30.5 U per mg of protein . The species could be characterized by their catalase and peroxidase since L . pneumophila and L . gormanii had only peroxidase (relative molecular weight {Mr}, 150,000); L . dumoffii had a peroxidase (Mr, 150,000) plus a catalase (Mr, 174,000); and all remaining species had catalase only (Mr, 300,000, 220,000, or 150,000).

Arch Biochem Biophys, 1984 Sep, 233(2), 643 - 51
Enolase isozymes from Ricinus communis: partial purification and characterization of the isozymes; Miernyk JA et al.; The plastid and cytosolic isozymes of enolase from developing endosperm of castor oil seeds, Ricinus communis L . cv . Baker 296, were separated and partially purified . Each purified isozyme had a specific activity of approximately 200 mumol min-1 mg protein . The isozymes have similar pH optima for the forward reaction, but different optima for the reverse reaction . The divalent metal specificity is the same for both isozymes . In addition to differences in charge, the isozymes can be distinguished by their different kinetic constants, thermostability, and sensitivity to fluoride inhibition . Antibodies against yeast enolase isozyme I cross-react with Ricinus plastid enolase but not with the cytosolic isozyme.

Jpn J Pharmacol, 1984 Sep, 36(1), 77 - 85
Pharmacological properties of a new anti-inflammatory compound, alpha-(3,5-di-tert-butyl-4-hydroxybenzylidene)-gamma-butyrolacto ne (KME-4), and its inhibitory effects on prostaglandin synthetase and 5-lipoxygenase; Hidaka T et al.; The pharmacological effects of a new anti-inflammatory compound, alpha-(3,5-di-tert-butyl-4-hydroxybenzylidene)-gamma-butyrolactone (KME-4), and its inhibitory effects on arachidonate prostaglandin synthetase and 5-lipoxygenase activities were examined . KME-4 showed anti-inflammatory activity . It was less active than indomethacin, but more active than naproxen and ibuprofen in carrageenin-induced paw edema in rats; and it was less active than indomethacin, equipotent as naproxen, but more active than ibuprofen in granuloma formation in rats . The ulcerogenic activity of KME-4 was weaker than indomethacin and naproxen, but stronger than ibuprofen in starved rats . The ratio of UD50 stomach to ED30 carrageenin edema or to ED25 granuloma for KME-4 showed higher values than those of the reference drugs . KME-4 showed antipyretic activity in yeast-induced fever in rats . It also inhibited platelet aggregation induced by arachidonic acid and protected rabbits from arachidonic acid-induced death . Furthermore, KME-4 was found to be equipotent in inhibiting both prostaglandin synthetase and 5-lipoxygenase activities of rat basophilic leukemia cells, unlike indomethacin, naproxen and ibuprofen . It also inhibited the prostaglandin synthetase activity of bovine seminal vesicle . The present findings indicate that KME-4 may be a new type of anti-inflammatory drug with dual prostaglandin synthetase and 5-lipoxygenase inhibition.

Mech Ageing Dev, 1984 Sep, 27(1), 1 - 13
Growth rate and life span in Drosophila . I . Methods and mechanisms of variation of growth rate; Economos AC et al.; It has been suggested that development and ageing may be linked and it has been shown, in Drosophila, under conditions of varying developmental temperature and larval crowding that the rate of development may be inversely related with the duration of adult life . In order to test this hypothesis systematically, precise methods were devised for varying, in Drosophila, either growth rate or each of its components, i.e . body weight and duration of development, while holding the other constant . These methods are described in the present paper . Moreover, we report studies that shed some light on the mechanisms underlying the effects of temperature and larval crowding on Drosophila development . The major novel findings from these studies were: (a) the restriction of the amount of yeast per larva as larval density increases accounts entirely for the effect of larval crowding on duration of development but only for about two-thirds of its effect on body size; and (b) the increased size of flies grown at lower temperatures may be due to assimilation of more food rather than to more efficient assimilation of food.

Biochem J, 1984 Sep 1, 222(2), 407 - 11
Oxidation of tyrosine residues in proteins by tyrosinase . Formation of protein-bonded 3,4-dihydroxyphenylalanine and 5-S-cysteinyl-3,4-dihydroxyphenylalanine; Ito S et al.; A simple and rapid method was developed for the determination of 3,4-dihydroxyphenylalanine (dopa) and 5-S-cysteinyl-3,4-dihydroxyphenylalanine (5-S-cysteinyldopa) in proteins with the use of high-pressure liquid chromatography . With this method, it is demonstrated that mushroom tyrosinase can catalyse hydroxylation of tyrosine residues in proteins to dopa and subsequent oxidation to dopaquinone residues . The dopaquinone residues in proteins combine with cysteine residues to form 5-S-cysteinyldopa in bovine serum albumin and yeast alcohol dehydrogenase, whereas dopa is the major product in bovine insulin, which lacks cysteine residues.

Mol Biol (Mosk), 1984 Sep-Oct, 18(5), 1181 - 93
{Enzymatic synthesis of tRNA fragments}; Zhenodarova SM et al.; In the present paper the results of enzymatic synthesis of yeast tRNA1Val fragments have been summarized . It is shown that complex use of nucleolytic enzymes is a convenient and effective method of synthesis of the defined sequence oligoribonucleotides . The consecutive use of different nucleolytic enzymes (ribonucleases with different substrate specificity and polynucleotide phosphorylase) and RNA ligase has permitted to obtain various fragments (or their analogs) of T psi-loop, D-arm, anticodon arm and acceptor stem . Some fragments containing modified nucleosides such as tetranucleotide GpDpCpGp (fragment 15-18), octanucleotide GpUpCpUpApGpDpC (analog of fragment 10-17), nonanucleotide GpTpUpCpGpApUpCpC (analog of T psi-loop), decanucleotide psi pCpUpGpCpUpUpIpApC (analog of fragment 27-36), hexanucleotide CpApCpGpCpA (fragment 36-41) and others were synthesized.

Nature, 1984 Aug 9-15, 310(5977), 514 - 6
Role of reverse transcription in the generation of extrachromosomal copia mobile genetic elements; Flavell AJ; The Drosophila genetic element copia is one of the best studied eukaryotic transposable sequences . Copia shares structural features with a wide variety of mobile elements in Drosophila, Lepidoptera, yeast and vertebrates, the last class being the retrovirus proviruses . Furthermore, retrovirus-like particles containing copia RNA have been isolated from Drosophila cells and extrachromosomal circular copias with structures closely resembling circular retrovirus proviruses have been isolated and cloned . Therefore, copia-like elements and retroviruses may be members of a class of mobile genetic elements existing throughout the eukaryotic kingdom . Consequently, there has been speculation that retroviruses evolved from transposable elements and, conversely, that copia-like elements transpose as retroviruses or retrovirus-like particles . To date, however, there has been no demonstration that copia RNA is reverse transcribed into copia DNA . The present report describes the isolation of linear extrachromosomal copias whose structure closely resembles the analogous retrovirus provirus linears and whose synthesis is unaffected by inhibitors of the cellular DNA polymerase responsible for chromosomal DNA replication.

Nippon Yakurigaku Zasshi, 1984 Aug, 84(2), 243 - 9
{Effects of anti-inflammatory drugs on the lame walking reaction in adjuvant-induced edematous rats}; Higuchi S et al.; Acute inflammatory paw edema of rats was formed by the injection of 0.5% Mycobacterium tuberculosis-liquid parraffin suspension into the hind paw, and then the pain threshold of the inflamed paw decreased . At that time, the rats showed a three-legged gait, namely, the lame walking reaction . The reaction was inhibited by acidic nonsteroidal anti-inflammatory drugs, e.g., indomethacin, ibuprofen and aspirin, inhibitors of prostaglandins biosynthesis, at a lower dose level than those in the Randall-Selitto test using yeast edematous rats and in the flection tests using adjuvant arthritic or silver nitrate arthritic rats . On the other hand, basic nonsteroidal anti-inflammatory drugs, e.g., tiaramide HC1, mepirizole and perisoxal citrate, not inhibitors of prostaglandins biosynthesis, were less potent than the acidic nonsteroidal anti-inflammatory drugs in the inhibition of the lame walking reaction . When prostaglandin E2 was injected into the inflamed paw, the inhibitory effects of acidic non-steroidal anti-inflammatory drugs on the reaction disappeared, but those of the basic nonsteroidal anti-inflammatory drugs didn't disappear . Bradykinin had no influence on the effects of both acidic and basic nonsteroidal anti-inflammatory drugs in the inhibition of the reaction . Analgesic evaluation with the lame walking reaction is more sensitive than with the Randall-Selitto or the flection methods . Morphine, pentazocine and acetaminophen inhibited the reaction, and these effects didn't disappear by the injection of prostaglandin E2 into the inflamed paw . These results suggest that prostaglandins play important roles in inflammatory pain, and the lameness test can serve as a new method for evaluating analgesics such as anti-inflammatory drugs and for investigating the mechanism of inflammatory pain.

Am Rev Respir Dis, 1984 Aug, 130(2), 317 - 20
Tissue morphology of Histoplasma capsulatum in acute histoplasmosis; Reynolds RJ 3rd et al.; Reports of histopathology in acute histoplasmosis are rare . A case of fulminating acute histoplasmosis with hematogenous dissemination is described in which Histoplasma capsulatum was identified in transbronchial biopsy . This report represents the first morphologic description of H . capsulatum within human pulmonary tissue in the early phase of acute histoplasmosis . The yeast forms observed were of typical size, but they were present within the alveoli and were budding . No tissue granulomata were noted . This unusual morphology is in sharp contrast to the classic description of the organism in tissues and presented diagnostic difficulties, which are discussed in this report . Hematogenous dissemination is considered to be an important part of the pathogenesis of the disease, but it is rarely documented during the symptomatic phase of acute histoplasmosis . Cultural documentation of hematogenous dissemination was obtained in this patient . These observations stress the importance of obtaining culture material from extrapulmonary sites early in the course of acute histoplasmosis when a specific diagnosis is necessary.

Am J Clin Pathol, 1984 Aug, 82(2), 243 - 6
Cerebral blastomycosis: an immunodiagnostic study; Tang TT et al.; Cerebral blastomycosis may simulate a brain tumor . Its diagnosis is sometimes very difficult . The morphologic identification of the fungus may be misleading because it shares some common features with many other dimorphic fungi . Culturing and conversion of the organism from mycelial phase to yeast phase are not always successful . Immunofluorescent staining of the biopsy tissue is useful in confirming the diagnosis . However, a combination of double immunodiffusion (DID) test and complement fixation (CF) test makes the diagnosis more accurate and reliable . The direct role of macrophages in defending the host against blastomycosis is illustrated by electron microscopy.

Arch Biochem Biophys, 1984 Aug 1, 232(2), 496 - 504
Hypothesis--a chemical mechanism for the biosynthesis of ATP involving ion-exchange reactions; Jenkins WT et al.; Dissociation constants for Mg . ATP were determined by displacing ATP from Dowex-1 resin with magnesium . These constants were then used to analyze the kinetics of yeast mitochondrial ATPase, in terms of the concentrations of free magnesium and free ATP, at a series of pH values . Both Mg . ATP and hydroxide ions were found to compete with the binding of ATP to the enzyme . These results were interpreted, in terms of an ion-exchange model, to mean that the synthesis of ATP may require the utilization of both magnesium and hydroxide ions for the dissociation of ATP from the enzyme as Mg . ATP . The concentrations of Mg and hydroxide required to compete with ATP were both found to be about three orders of magnitude greater than those required to form products, indicating that magnesium and hydroxide ions can contribute about 8 kcal of energy when ATP is synthesized.

J Biomol Struct Dyn, 1984 Aug, 2(1), 55 - 61
Mechanistic studies on metal-pyrophosphate hydrolysis . Structure of tetraammine(chloroaquo)cobalt (III) dichloride ({Co(NH3)4ClH2O}2+.2Cl-), an acid hydrolysis product of Co(NH3)4HP2O7 and Co(NH3)4PO4; Sundaralingam M et al.; The octahedral complex tetraammine(chloroaquo)cobalt(III) dichloride is shown to be the HCl hydrolysis product of both P1,2-bidentate tetraammine(pyrophosphato)cobalt(III) {Co(NH3)4HP2O7 or CoPP} and bidentate tetraammine(phosphato)cobalt(III) {Co(NH3)4PO4 or CoP} . The complex crystallizes in the orthorhombic space group Pna21 with cell dimensions a = 13.033(2)A, b = 6.710(1)A, and c = 10.318(2)A; the crystal structure was refined to a final disagreement index of 0.033 . The average of the four Co-N distances is 1.944 +/- 6A . The Co-Cl distance is 2.257(2)A and the Co-O(W) distance is 1.971(4)A . Both protons of the coordinated water molecule are engaged in strong hydrogen bonds to the two nonbonded chloride counterions with O(W)-Cl distances of 3.087(6)A and 3.123(6)A . Each nonbonded chloride is engaged in seven hydrogen bonding interactions resulting from the high ratio of hydrogen bond donors to acceptors in the CoP structure . Cobalt bisphosphate (CoP2) is the final enzyme hydrolysis product when CoPP is used as substrate in the yeast inorganic pyrophosphatase reaction . The bridge oxygen atom is the site of initial CoPP cleavage both for HCl catalyzed hydrolysis as well as for enzyme catalyzed hydrolysis.

J Biomol Struct Dyn, 1984 Aug, 2(1), 165 - 74
An unexpected major groove binding of netropsin and distamycin A to tRNA(phe); Rubin J et al.; Crystalline complexes of yeast tRNA(phe) and the oligopeptide antibiotics netropsin and distamycin A were prepared by diffusing drugs into crystals of tRNA . X-ray structure analyses of these complexes reveal a single common binding site for both drugs which is located in the major or deep groove of the tRNA T-stem . The netropsin-tRNA complex is stabilized by specific hydrogen bonds between the amide groups of the drug and the tRNA bases G51 O(6), U52 O(4) and G53 N(7) on one strand, and is further stabilized by electrostatic interactions between the positively charges guanidino side chain of the drug and the tRNA phosphate P53 on the same strand and the positively charged amidino propyl side chain and the phosphates P61, P62 and P63 on the opposite strand of the double helix . These results are in contrast to the implicated minor groove binding of these drugs to non-guanine sequences in DNA . The binding to the GUG sequence in tRNA implies that major groove binding to certain DNA sequences is possible.

Nucleic Acids Res, 1984 Jul 25, 12(14), 5757 - 65
A leucine tRNA gene adjacent to the QA gene cluster of Neurospora crassa; Huiet L et al.; A single tRNALeu gene has been localized and sequenced from Neurospora crassa . It is located only 375 bp from the qa gene cluster and it is the only tRNA or 5S rRNA gene within this cloned 37 kb region . The gene encodes a tRNALeu with the anti-codon AAG, and unlike the other nuclear eukaryotic tRNALeu (AAG) gene sequenced (from C . elegans), contains an intervening sequence of 27 bp . The Neurospora tRNALeu (AAG) is 84% and 73% homologous respectively to the C . elegans and bovine tRNALeu (AAG), and is 84% homologous to a Drosophila tRNALeu (CAA) . However, it is only 65% homologous to a yeast tRNALeu (CAA) and there is little conservation of intervening sequences or V-loop regions . The gene hybridizes to at least 16 other DNA fragments in the Neurospora genome . Its expression does not seem to be linked to that of the qa genes.

Nucleic Acids Res, 1984 Jul 25, 12(14), 5737 - 55
Accurate transcription of homologous 5S rRNA and tRNA genes and splicing of tRNA in vitro by soluble extracts of Neurospora; Tyler BM et al.; We have developed soluble extracts from Neurospora crassa capable of accurately and efficiently transcribing homologous 5S rRNA and tRNA genes . The extracts also appear to quantitatively end-process and splice the primary tRNA transcripts . Although the extracts could not transcribe a heterologous (yeast) 5S rRNA gene, they did transcribe a yeast tRNALeu gene and slowly process the transcripts . In addition, we have developed a novel strategy for rapidly sequencing uniformly labelled RNAs using base-specific ribonucleases . We have used this procedure to verify the identity of the in vitro transcripts and processing products.

Biochemistry, 1984 Jul 17, 23(15), 3501 - 7
Photoaffinity labeling of the major nucleosidetriphosphatase of rat liver nuclear envelope; Clawson GA et al.; We employed the photoaffinity probe 8-azido-adenosine 5'-triphosphate (aATP) to identify the nuclear envelope (NE) nucleosidetriphosphatase activity (NTPase) implicated in control of RNA transport . The photoprobe was hydrolyzed at rates comparable to those for ATP, with a Michaelis constant of 0.225 mM . Photolabeling was dependent upon UV irradiation (300-nm max) and was not affected by quercetin . Unlabeled ATP or GTP competed with {32P}aATP in photolabeling experiments, and UTP was a less effective competitor, paralleling the substrate specificity of the NTPase . Incubation of NE with aATP led to a UV, time, and concentration dependent irreversible inactivation of NTPase . The inactivation could be blocked by ATP or GTP . Polyacrylamide gel electrophoresis and autoradiography of photolabeled NE showed selective, UV-dependent labeling of a 46-kDa protein with both {gamma-32P}aATP and {alpha-32P}aATP . This band was not labeled with {gamma-32P}ATP . Since the NE NTPase implicated in RNA transport is modulated by RNA, we examined the effects of RNA on the labeling process . Removal of RNA from the NE preparations (by RNase/DNase digestion) reduced NTPase by 30-40% and eliminated photolabeling of the 46-kDa band . Addition of yeast RNA to such preparations increased NTPase activity to control levels and selectively reinstated photolabeling of the 46-kDa band . These results suggest that the 46-kDa protein represents the major NTPase implicated in RNA transport.

Nucleic Acids Res, 1984 Jul 11, 12(13), 5515 - 27
Coding strategy differences between constant and variable segments of immunoglobulin genes; Perrin P; Vertebrate immunoglobulin (Ig) mRNAs reveal intraspecies variation in codon usage distinct from that seen with yeast or bacterial genes . Comparison of all available Ig gene sequences shows that %(G + C) in codon position III is consistently lower in variable (V) segments than in constant (C) segments . I find an even lower %(G + C) in the hypervariable domains of V segments . This analysis suggests that base substitution in Ig genes correlates positively with local A + T content.

J Clin Pathol, 1984 Jul, 37(7), 783 - 6
Defective phagocytosis in insulin controlled diabetics: evidence for a reaction between glucose and opsonising proteins; Davidson NJ et al.; Neutrophils from diabetic patients controlled with insulin showed impaired phagocytosis of a yeast, Candida guilliermondii . The defect was detected by measuring the initial rate of phagocytosis at optimal concentrations of phagocytes and organisms in autologous plasma . By mixing normal neutrophils in diabetic plasma and vice versa both cellular and plasma abnormalities were shown . The defect was reproduced by incubating normal plasma at a D-glucose concentration of 15 mmol/l for 3 h or for shorter periods at higher concentrations of glucose . The data suggest that defective phagocytosis is partly due to a reaction between glucose and the plasma proteins concerned with opsonisation . Defective phagocytosis may be important not only in coping with infections but also in other diabetic complications as plasma proteins are concerned with the removal of damaged or effete cells as well as foreign antigens.

J Exp Med, 1984 Jul 1, 160(1), 197 - 207
Accumulation of diabetic rat peripheral nerve myelin by macrophages increases with the presence of advanced glycosylation endproducts; Vlassara H et al.; We have previously shown that increased nonenzymatic glycosylation occurs in peripheral nervous tissue of diabetic humans and animals, primarily on the PO-protein of peripheral nerve myelin . The pathophysiologic mechanism by which this biochemical alteration leads to myelin breakdown and removal is not as yet understood . In the present study we show that advanced glycosylation end-product (AGE) adducts that form during long-term exposure of peripheral nerve myelin proteins to glucose in vitro and in vivo markedly alter the way in which myelin interacts with elicited macrophages . In this interaction, macrophages appear to specifically recognize AGEs on myelin, since AGE-BSA competes nearly as effectively as AGE-myelin, while neither unmodified BSA nor unmodified myelin compete . The failure of yeast mannan to interfere with macrophage recognition of AGE-myelin suggests that the mannose/fucose receptor does not mediate this process . Recognition of AGE-protein by macrophages is associated with endocytosis, as demonstrated by resistance of cell-associated radioactivity to removal by trypsin action, and by low temperature inhibition of ligand accumulation in the cellular fraction . 125I-labeled myelin that had been incubated in vitro with 50 mM glucose for 8 wk reached a steady state accumulation within thioglycolate-elicited macrophages that was five times greater than that of myelin incubated without glucose . Similarly, myelin isolated from rats having diabetes for 1.5-2.0 years duration had a steady state level that was 9 times greater than that of myelin from young rats, and 3.5 times greater than that of myelin from age-matched controls . In contrast, myelin isolated from rats having diabetes for 4-5 wk had the same degree of accumulation observed with myelin of age-matched normal rats . These data suggest that the amount of increased nonenzymatic glycosylation observed in the myelin of short-term diabetic rats had not yet resulted in the significant accumulation of AGE-myelin present both in vitro and in the long-term diabetic rats . The disappearance of acid-insoluble radioactivity from within the cells and the appearance of acid-soluble radioactivity released into the medium were very similar for the two groups, suggesting that the striking difference in accumulation seen between normal myelin and AGE-myelin is due primarily to increased uptake . Formation of irreversible AGE-adducts on myelin appears to promote the recognition and uptake of the modified myelin by macrophages . This interaction between AGE-myelin and macrophages may initiate or contribute to the segmental demyelination associated with diabetes and the normal aging of peripheral nerve.

J Cell Physiol, 1984 Jul, 120(1), 91 - 102
Effect of retinoic acid on the proliferation and phagocytic capability of murine macrophage-like cell lines; Goldman R; Retinoic acid (RA) exerted a variable degree of growth inhibitory activity on the macrophage-like cell lines P388D1, J774.2, WEHI-265, WEHI-3, and PU-5 . Comparison of cell proliferation and clonal growth suggests that at concentrations of 10(-9)-10(-6) M the inhibitory activity stems from processes leading to elongation of cell cycle time and not from terminal differentiation processes . RA was shown to be a potent inducer of the development of high-phagocytic phenotypes (assessed by phagocytosis of heat-killed yeast cells) in the P388D1, J774.2, and WEHI-265 cell lines which differ substantially in their proliferative and adherence characteristics . The PU-5 and WEHI-3 cell lines were not induced by RA to express an enhanced phagocytic activity toward heat-killed yeast cells . The augmented phagocytic capability was dose dependent over a wide range of RA concentrations . In P388D1 cells, 2 X 10(-12) M RA already exerted significant phagocytosis augmentation effects, which progressively increased up to 2 X 10(-5) M RA, the highest concentration tested . Retinal, retinyl acetate, and retinol had similar effects to those of RA on both cell adherence and phagocytosis in P388D1 cells, albeit at concentrations four to six orders of magnitude higher . Optimal development of the high-phagocytic phenotype in P388D1, J774.2, and WEHI-265 cells required at least 96 hr of culture in the presence of RA; at 48 hr and 23 hr the effects were already substantial, whereas at 4 hr of exposure to RA no significant enhancement of phagocytosis could be detected . Thus both extended periods of culture in the presence of RA (more than two to three cell cycles) and high concentrations were needed for induction, in more than 90% of the cells, of the expression of a high-phagocytic phenotype . The reversion to a low-phagocytic phenotype upon removal of RA was also rather slow and required several cell cycles . In P388D1 cells RA also enhanced the phagocytosis of latex beads but had no effect on the phagocytosis of starch particles, or the extent of binding of immunoglobulin G-coated sheep red blood cells (SRBC) . The expression of receptors for concanavalin A and for nonopsonized SRBC remarkably increased in RA-treated cells, as was the ability to perform Fc-receptor mediated erythrophagocytosis . Both P388D1 cells and WEHI-265 cells were induced by RA to express nitroblue tetrazolium reducing activity . The data suggest that RA induces profound changes in the functional capabilities of macrophage-like cell lines which are apparently not dependent on cessation of growth and terminal differentiation processes.

J Bacteriol, 1984 Jul, 159(1), 353 - 9
Properties of salicylate hydroxylase and hydroxyquinol 1,2-dioxygenase purified from Trichosporon cutaneum; Sze IS et al.; Salicylate hydroxylase (salicylate 1-monooxygenase, EC 1.14.13.1) was purified from the soil yeast Trichosporon cutaneum . The enzyme contained flavin adenine dinucleotide and was monomeric, with a molecular weight of 45,300 . In addition to salicylate, the four isomeric dihydroxybenzoates having one hydroxyl adjacent to carboxyl in the benzene nucleus were oxidatively decarboxylated without formation of hydrogen peroxide . One of these isomers, gentisate, was rapidly oxidized to hydroxyquinol by the enzyme but did not serve as an effective single carbon source for T . cutaneum; however, when growing with salicylate, cells also readily utilized gentisate for growth . Hydroxyquinol 1,2-dioxygenase (EC 1.13.11....) is a newly investigated enzyme which was purified from T . cutaneum grown with 4-hydroxybenzoate . The enzyme was red, contained ferric iron, and was specific for hydroxyquinol; catechol and pyrogallol were oxidized at less than 1% of the rate for hydroxyquinol, and no activity could be detected against seven other catechols . The enzyme was composed of two nonidentical subunits having molecular weights of 39,600 and 38,200 and was apparently dimeric.

J Bacteriol, 1984 Jul, 159(1), 390 - 2
Ca(II)-calmodulin regulation of fungal dimorphism in Ceratocystis ulmi; Muthukumar G et al.; We have shown that Ca(II) ions, ethylene glycol-bis(beta-aminoethyl ether)-N,N-tetraacetic acid, LaCl3, and six known calmodulin inhibitors shift the yeast-mycelium dimorphic potential of Ceratocystis ulmi . Our data are consistent with the conclusions that Ca(II)-calmodulin interaction is necessary for mycelial growth in C . ulmi and that the absence of this interaction leads to the yeast phase.

Antibiotiki, 1984 Jul, 29(7), 490 - 5
{Interrelation between the activity of the lytic enzyme producer Actinomyces recifensis var . lyticus 2435 and its population variability}; Babenko IuS et al.; Spontaneous variability of Actinomyces recifensis var . lyticus 2435 producing a complex of lytic enzymes was studied . A correlation between the strain activity and the content of the variants of the main morphological type in the population was shown . The carbon sources influenced the proportion of different type variants in the population . Strain 2435 was rather stable with respect to the level of the synthesis of yeast-like enzymes and showed a significant variation with respect to the level of the biosynthesis of the bacteriolytic complex . The population variation of strain 2435 with respect to the staphylolytic (synthesis of specific endopeptidases) and muramidase activity was most pronounced . The high activity levels of strain 2435 and wide lytic spectrum were provided by selection of the variants of the first morphological type with respect to the property of high staphylolytic activity.

Zh Mikrobiol Epidemiol Immunobiol, 1984 Jul, (7), 26 - 9
{Various problems of biotechnology}; Zhdanov VM; The prospects of using the methods of gene engineering for the development of viral vaccines are considered . The technological and biological barriers hampering the introduction of these biotechnological methods into the practice of vaccinal prophylaxis are analyzed . The work points out that in the near future successes may be achieved in synthetizing the internal proteins M and NP of influenza viruses and similar proteins of paramyxoviruses and rhabdoviruses, as well as in using yeast for the production of vaccine against hepatitis B.

EMBO J, 1984 Jul, 3(7), 1581 - 5
Two different types of intervening sequences in the glucoamylase gene from Aspergillus niger; Boel E et al.; One single glucoamylase gene could be identified in the chromosomal DNA of Aspergillus niger by Southern blot analysis . This glucoamylase gene was isolated from a genomic library of A . niger DNA . The glucoamylase gene is situated on a 2.5-kb EcoRI-EcoRV fragment and contains five intervening sequences in the coding region . One 169-bp intron is involved in differential mRNA processing leading to the two different glucoamylase enzymes G1 and G2; the other four introns are all very small ranging from 55 to 75 bp in length . One intron has a significant homology to the coding region which immediately follows, and it contains the internal conserved sequence TACTAAC, which is also found in yeast chromosomal gene introns, and is thought to participate in mRNA splicing . Two transcription initiation sites and a typical eukaryotic promoter region with TATAAT and CAAT boxes are located upstream from the gene.

Cell, 1984 Jul, 37(3), 915 - 25
A minimal intron length but no specific internal sequence is required for splicing the large rabbit beta-globin intron; Wieringa B et al.; We constructed rabbit beta-globin genes with deletions in the large intron, extending from the midpoint toward the 5' or 3' splice sites . Analysis of transcripts in transformed HeLa cells showed that six 5' proximal intron nucleotides allowed normal splicing . Correct splicing at the 3' splice site required 12 or more 3' proximal intron nucleotides; optimal efficiency required 24 nucleotides . Remarkably, a mini-intron comprising six 5' and 24 3' intron nucleotides gave no correctly spliced transcripts; extending the miniintron with polyoma or pBR322 fragments to 80 or more nucleotides restored normal splicing . Thus other than in yeast nuclear genes, no specific internal intron sequences appear to be needed but a minimal intron length is important.

Toxicol Appl Pharmacol, 1984 Jun 30, 74(2), 293 - 5
Pyrvinium pamoate lacks in vivo genotoxicity in the colon; Goldberg MT; Pyrvinium pamoate, a widely used anthelmintic drug, has been previously tested for genotoxicity with equivocal results . Assays with bacterial or yeast test strains yielded positive results while in vitro tests with mammalian cells yielded negative results . In this study, the genotoxicity of pyrvinium was studied in vivo in the mouse colon, the therapeutic site of action of this agent . 1,2-Dimethylhydrazine (DMH), a colon carcinogen, was tested simultaneously as a positive control . The colonic nuclear aberration assay was used to determine genotoxicity . Pyrvinium delivered orally in three vehicles was not genotoxic in the murine colon, even at doses up to 12.5 times the recommended human dosage.

Mycopathologia, 1984 Jun 30, 86(3), 185 - 90
Case report: blastomycosis causing sciatic neuritis; Miller-Catchpole R et al.; A patient presented with pain in the right lower back, radiating down the right leg . Initial pelvic X-rays did not reveal any lesion . A follow up computerized tomography (CT) scan and technitium scan showed a sharply lytic lesion of the right ilium extending to the right sacroiliac joint . Open biopsy revealed a granulomatous inflammation with many budding yeast from consistent with Blastomyces dermatitidis . Subsequent culture confirmed this identification . There was no other site of fungal infection . Two courses of Amphotericin B (each to 2 g total dose) failed to eradicate this infection . The patient is now responding to Ketoconazole.

J Biol Chem, 1984 Jun 25, 259(12), 7835 - 41
Phosphorylation sites in enolase and lactate dehydrogenase utilized by tyrosine protein kinases in vivo and in vitro; Cooper JA et al.; Enolase, lactate dehydrogenase, and phosphoglycerate mutase have previously been found to contain phosphotyrosine in fibroblasts transformed by Rous sarcoma virus, which encodes a tyrosine-specific protein kinase . However, these phosphorylations are not stoichiometric, and their significance for any aspect of the transformed phenotype is unknown . We show here that enolase and lactate dehydrogenase are each phosphorylated chiefly at a single tyrosine in Rous sarcoma virus-transformed cells . The purified enzymes can also be phosphorylated at the same tyrosine in vitro when incubated with an immunoprecipitated retroviral transforming protein having associated tyrosine protein kinase activity . The phosphorylated tyrosine in lactate dehydrogenase is amino acid 238 . The phosphorylated tyrosine in enolase lies in a sequence homologous to that surrounding histidine 43 in yeast enolase . Although the phosphorylated sequence in lactate dehydrogenase shows some homology to those sequences surrounding phosphotyrosines found in retroviral transforming proteins, the phosphorylated sequence in enolase is quite different.

Neurosci Lett, 1984 Jun 15, 47(2), 113 - 8
Behavioral evidence in rats for a peptidergic-noradrenergic interaction in cutaneous sensory and vascular function; Coderre TJ et al.; Cutaneous sensory and vascular function was examined following application of capsaicin to the sciatic nerve and systemic injection of guanethidine . Together the two drugs produced a reduction in sensitivity to heat-pain, inflammatory pain (formalin test), tactile stimulation and skin temperature of the foot that exceeded the effects of either drug alone . The inflammation produced by an injection of formalin to the plantar surface of the hind paw was reduced equally by capsaicin or capsaicin + guanethidine . Cold sensitivity and inflammation produced by yeast injection were unaffected by all treatments . The data imply a peripheral interaction between peptidergic and noradrenergic systems with significant functional implications that may be important in the pathology of familial dysautonomia.

Biochim Biophys Acta, 1984 Jun 6, 794(1), 96 - 103
Nonspecific inhibition of enzymes by p-bromophenacyl bromide . Inhibition of human platelet phospholipase C and modification of sulfhydryl groups; Kyger EM et al.; This study demonstrates that p-bromophenacyl bromide irreversibly inhibits, in a time- and dose-dependent manner, yeast alcohol dehydrogenase, bovine pancreatic alpha-chymotrypsin, human platelet phosphatidylinositol (PI)-specific phospholipase C, in addition to the neutral-active and calcium-dependent phospholipase A2 of human platelets . The PI-specific phospholipase C has maximal activity at pH 5,5 is calcium-dependent, and is strongly inhibited by sulfhydryl reagents 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) and methylmethane thiosulfonate . Increasing concentrations of DTNB produced concomitant inhibition of phospholipase C activity and titration of sulfhydryl groups . In contrast, human platelet phospholipase A2 activity was unaffected by concentrations of DTNB that titrated sulfhydryl groups, and completely inhibited PI-specific phospholipase C activity . Treatment of cysteine with p-bromophenacyl bromide resulted in modification of the amino acid as demonstrated by paper chromatography, and loss of titratable sulfhydryl groups . These data show that p-bromophenacyl bromide inhibits a wide spectrum of enzymatic activities including PI-specific phospholipase C . This reagent modifies amino acid residues other than active-site histidines and therefore has a broader reactivity than previously considered . Thus, it should not be used as a selective inhibitor of enzymes in crude cellular experiments.

Proc Natl Acad Sci U S A, 1984 Jun, 81(11), 3361 - 4
Signal recognition particle is required for co-translational insertion of cytochrome P-450 into microsomal membranes; Sakaguchi M et al.; Insertion of newly synthesized P-450(1), the major phenobarbital-inducible form of rabbit liver microsomal cytochrome P-450, into microsomal membranes was studied in a wheat germ cell-free translation system programed with total RNA from the liver of a phenobarbital-treated rabbit . P-450(1) synthesized in vitro had the same molecular weight as the mature molecule and was co-translationally inserted into dog pancreas rough microsomal membranes . In the presence of salt-washed microsomes, instead of unwashed ones, the insertion was greatly diminished . It could, however, be restored by supplementation of the system with purified signal recognition particle (SRP), a known component of the membrane translocation machinery for secretory proteins . In the absence of microsomes, SRP inhibited the translation of mRNA encoding P-450(1) and this translation arrest was released by the addition of salt-washed microsomes . On the other hand, SRP did not affect the translation of mRNAs encoding yeast porin and reticulocyte globin, which are mitochondrial and cytosolic proteins, respectively . We conclude that co-translational insertion of P-450(1) into microsomal membranes requires SRP and postulate that P-450(1) possesses an uncleavable signal sequence that can be recognized by SRP.

J Leukoc Biol, 1984 Jun, 35(6), 605 - 15
Enhanced phagocytic activity of blood leukocytes in athymic nude mice; Holub M et al.; After 30-min incubation, blood leukocytes of adult athymic BALB/c nude mice (nu/nu) have a distinctly higher methacrylate or yeast particle uptake than leukocytes of euthymic nu/+ or +/+ mice . This permanent enhancement is not due to humoral factors, since the percentage of phagocytosing nu/nu leukocytes increases further in nu/+ littermate's plasma . Also, chronic infection or intraperitoneal immunization causes an additional transitory increase of the percentage of phagocytosing leukocytes and numbers of particles ingested; the phagocytic performance of leukocytes of euthymic mice is raised under these conditions by a greater factor . In fetuses enhanced phagocytosis of nu/nu mice is found only in monocytes . The bulk of ingestion of methacrylate particles is mediated by Fc receptors and significantly more receptors for IgG2B, IgG1, and IgG2A are demonstrable on nu/nu neutrophils; ingestion of particles via these receptors is again higher in nu/nu neutrophils, whereas nu/nu monocytes display a higher uptake via Fc(IgG1) receptors.

Agents Actions, 1984 Jun, 14(5-6), 654 - 61
The antiinflammatory profile of (5H-dibenzo{a,d}-cyclohepten-5-ylidene)acetic acid (WY-41,770), an agent possessing weak prostaglandin synthetase inhibitory activity that is devoid of gastric side effects; Carlson RP et al.; Wy-41,770 {(5H-dibenzo{a,d}cyclohepten-5-ylidene)acetic acid}, a novel acrylic acid, was compared to indomethacin and aspirin in standard antiinflammatory, analgesic and antipyretic animal models . The acute antiinflammatory, analgesic and antipyretic activity of Wy-41,770 (oral ED50S 50-170 mg/kg) was similar to aspirin; however, it was considerably more potent orally in adjuvant arthritis in the rat (ED50, 16 mg/kg) and urate-induced synovitis in the dog (ED50, 4.5 mg/kg) . Wy-41,770 was a weak inhibitor of prostaglandin biosynthesis and did not inhibit either 5- or 15-lipoxygenase . Furthermore, the cellular migration characteristic of carrageenan pleurisy was not affected by Wy-41,770 . Unlike a majority of NSAIDs, it produced no gastric irritation in rats after either acute or chronic oral administration over the range 400-800 mg/kg . The major mechanism of action of Wy-41,770 has yet to be identified but does not seem to involve interference of arachidonic acid metabolism.

Nippon Yakurigaku Zasshi, 1984 Jun, 83(6), 513 - 21
{Effect of various types of analgesic drugs on an experimental model of chronic inflammatory pain in mice}; Okuyama S et al.; We attempted to develop an experimental model of chronic inflammatory pain in mice . The mice were injected intradermally at the base of the tail with various kinds of irritants (yeast, carrageenin, mustard and adjuvant) . The pain threshold was measured by the pressure method every 60 min for 5 hr and once a day at the same time throughout the experimental period . The group of 10% yeast-injected mice exhibited the most intensive hyperalgesia . The analgesic effect of various types of analgesic drugs were studied, comparing the effects in normal mice and various kinds of irritants-induced hyperalgesia mice . It was demonstrated by observing ED50 values that nonsteroidal antiinflammatory drugs (NSAIDs), narcotic analgesic drugs and agonist/antagonist type of analgesic drugs were effective, but CNS-acting drugs were ineffective in yeast hyperalgesia mice . In comparison with yeast hyperalgesia mice, larger doses of analgesic drugs were required in normal mice and other irritants-treated mice . Especially, acidic NSAIDs were more effective in yeast hyperalgesia mice than normal mice . It was suggested that acidic NSAIDs specifically inhibit inflammatory pain . Moreover, yeast hyperalgesia mice are useful for the quantitative measurement of analgesic drugs.

Mycopathologia, 1984 May 30, 86(2), 103 - 11
Phialophora richardsiae isolated from infected human bone: morphological, physiological and antifungal susceptibility studies; Yangco BG et al.; A dematiaceous fungus, Phialophora richardsiae (Nannf.) Conant, was isolated from human bone . In culture the fungus produced no yeast forms and was less pigmented than two other P . richardsiae isolates . While growth rates were similar, colonial forms differed . Phialides were of two kinds . While both had broad bases and tapered at the tips, only one terminated with a cupulate or rarely a saucer-shaped collarette . Most phialides were hyaline with a few lightly pigmented ones in older cultures . Broth dilution susceptibility testing of the isolates against amphotericin B, miconazole, ketoconazole, clotrimazole, and 5-fluorocytosine showed the fungus was susceptible to miconazole, ketoconazole and amphotericin B at achievable serum levels and resistant to 5-fluorocytosine and clotrimazole . The other isolates were reported to differ in their resistance to miconazole and amphotericin B . Enzyme and salinity studies showed minor difference among the isolates.

J Biol Chem, 1984 May 25, 259(10), 6090 - 7
Pyrophosphate of high and low energy . Contributions of pH, Ca2+, Mg2+, and water to free energy of hydrolysis; de Meis L; The equilibrium between inorganic pyrophosphate and inorganic orthophosphate was determined at pH values varying between 6.0 and 8.0, in the presence of different concentrations of MgCl2, mixtures of MgCl2 and CaCl2, and different organic solvents . The reactions were catalyzed by yeast inorganic pyrophosphatase . It was found that at 35 degrees C, depending on the conditions used, the observed equilibrium constant of pyrophosphate hydrolysis vary from a value higher than 4 X 10(3) M (delta Goobs more negative than -5.1 kcal/mol) to a value as low as 3 M (delta Goobs -0.7 kcal/mol) . The experimental data were used to compute the equilibrium constants of the reactions involving different ionic species . The data presented are interpreted according to the concept that the Keq of hydrolysis of a high energy compound depends on the difference in solvation energy of reactants and products.

Nucleic Acids Res, 1984 May 11, 12(9), 4001 - 9
Nucleotide sequence of the 3'-terminal tRNA-like structure in barley stripe mosaic virus genome; Kozlov YuV et al.; This paper describes the sequence of 257 nucleotides from the 3' end of RNA 2 of barley stripe mosaic virus ( BSMV , strain Argentina Mild) including an internal oligo (A) tract localized at a distance of 236 nucleotides from the 3' end, and the 3' terminal tRNA-like structure accepting tyrosine . This sequence is shown to be the same with RNAs 1,2 and 3 of another BSMV strain, Norwich , for at least the first 106 nucleotides from the 3' end . The 3' extremity of BSMV RNA bears some resemblance to tRNATyr from yeast in its primary structure . The possible secondary structures of the tRNA-like sequence in BSMV genome are discussed.

Nucleic Acids Res, 1984 May 11, 12(9), 3983 - 4000
Molecular analysis of the alcohol dehydrogenase (Adh1) gene of maize; Dennis ES et al.; A cDNA clone of maize Adh1 which contains the entire protein coding region of the gene has been constructed . The protein sequence predicted from the nucleotide sequence is in agreement with limited protein sequencing data for the ADH1 enzyme . An 11.5 kb genomic fragment containing the Adh1 gene has been isolated using the cDNA clone as a probe, and the gene region fully sequenced . The gene is interrupted by 9 introns, their junction sequences fitting the animal gene consensus sequence . Within the gene there is a triplication of a segment (104 bp) spanning an intron-exon junction . Presumptive promoter elements have been identified and are similar in nucleotide sequence and location, relative to the start of transcription, to those of other plant and animal genes . No recognizable poly(A+) addition signal is evident . Comparison of the nucleotide sequences of the cDNA (derived from an Adh1 -F allele) and genomic (derived from an Adh1 -S allele) clones has identified an amino acid difference consistent with the observed difference in electrophoretic mobility of the two enzymes . The maize ADH1 amino acid sequence is 50% homologous to that of horse liver ADH but is only 20% homologous to yeast ADH.

Biochemistry, 1984 May 8, 23(10), 2139 - 44
Bovine conglutinin is a collagen-like protein; Davis AE 3rd et al.; Conglutinin is a bovine plasma protein which is relatively large and asymmetric with elevated contents of glycine and, to some extent, proline . Although its physiologic function is unknown, conglutinin is known to bind, in the presence of calcium, to yeast cell walls and to the solid-phase-inactivated third component of complement . On sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions, isolated conglutinin appeared to consist of a single polypeptide chain (Mr 48 000) . Unreduced conglutinin consisted of a single stained band with an apparent molecular weight of approximately 300 000 . Cross-linking experiments with glutaraldehyde and dimethyl suberimidate suggested that this Mr 300 000 molecule consists of six of the disulfide-linked polypeptide chains . Amino acid composition revealed hydroxylysine and hydroxyproline together with elevated glycine and proline contents . Digestion of reduced, alkylated conglutinin with bacterial collagenase resulted in formation of a precipitate which consisted of an Mr 24 000 peptide which was digested to Mr 21 000 with large quantities of collagenase . These peptides contained less glycine, proline, hydroxylysine, and hydroxyproline than did the intact protein . The supernatant from this digestion mixture was, however, enriched in these four amino acids, with glycine making up nearly one-third of the total . Prolonged digestion with pepsin at 37 degrees C resulted in an Mr 20 000 peptide which was enriched in glycine, proline, hydroxyproline, and hydroxylysine . Amino-terminal sequence analysis showed that the glycine-X-Y repeating sequence begins at residue 26.(ABSTRACT TRUNCATED AT 250 WORDS)

FEBS Lett, 1984 May 7, 170(1), 162 - 4
Uric acid substantially enhances the free radical-induced inactivation of alcohol dehydrogenase; Kittridge KJ et al.; Lactate dehydrogenase (LDH) and yeast alcohol dehydrogenase ( YADH ) are inactivated when attacked by hydroxy free radicals (OH) . Organic molecules with a high rate constant of reaction with OH such as ascorbate or urate can compete with the enzymes for these strongly oxidising radicals . However, although 10(-3)M ascorbate can substantially protect both LDH and YADH from OH attack, in the presence of 10(-3)M urate only LDH is protected . In the case of YADH an even greater degree of inactivation than with OH occurs . The extent of inactivation is considerably reduced when oxygen is absent, in agreement with a urate peroxy radical perhaps being partly responsible for the increased inactivation of the enzyme.

Radiobiologiia, 1984 May-Jun, 24(3), 364 - 7
{Modeling the processes of cell recovery from radiation injury and the effective dose reduction principle . Model of postradiation recovery with a finite number of recovery channels}; Andreev AD; Using the effective dose reduction principle for the description of the postirradiation recovery a mathematical model has been developed with the definite number of recovery channels in a cell . It was established that the number of recovery channels in Megri 139-B yeast cells was of the order of 10 . The model may be applied for co-ordination of studies in the repair of radiation-induced lesions in a DNA molecule and in the postirradiation survival of exposed cells.

Clin Exp Immunol, 1984 May, 56(2), 321 - 9
Receptor associated defects of cultured monocytes in bronchial carcinoma; Lukacs K et al.; Using peripheral blood monocytes from individuals with bronchial carcinoma (BC) we have measured changes, over an 8 day culture period, in monocyte/macrophage complement (C3b) and IgG (Fc) rosettes, chemotactic factor (CF)-induced enhancement of C3b and IgG (Fc) rosettes, and the phagocytic index (PI) for yeast particles . The same measurements were made on monocyte/macrophages from individuals with non-malignant respiratory diseases (NMRD) and healthy controls (HC) . Following 8 days of culture there was a significant increase in C3b rosettes, IgG (Fc) rosettes and the PI in NMRD and HC . In contrast there was no significant increase in C3b rosettes in BC . The PI increased in BC, but on day 8 was still significantly less than control monocytes/macrophages . IgG (Fc) rosettes increased in BC following culture to a similar degree as that observed with NMRD and HC . In addition, there were no differences between BC and HC in CF-induced enhancement of IgG (Fc) rosettes at either day 0 or day 8 of culture whereas complement receptor enhancement in BC was impaired both at the beginning and end of the 8 day culture period . These results indicate that in advanced BC there is an abnormality in the expression of monocyte/macrophage complement receptors (which is not readily demonstrable for IgG {Fc}receptors) and that the defect persists following an 8 day culture period.

Allergy, 1984 May, 39(4), 259 - 74
A reference allergen preparation of the house dust mite D . pteronyssinus, produced from whole mite culture--a part of the DAS 76 study . Comparison with allergen preparations from other raw materials; Lind P et al.; A D . pteronyssinus whole culture allergen preparation contained 49 antigens as revealed by crossed immunoelectrophoresis (CIE), using polyspecific rabbit antibodies . Crossed radio-immunoelectrophoresis ( CRIE ) with sera from 30 patients revealed nine allergens, antigens 42, X, Y and 23 (in rank order) showing the most frequent and intense IgE-uptake . Nine antigens originated from the culture medium (human dander + yeast), but none of these gave rise to specific IgE-uptake . Extremely few and weak reactions were observed in radioallergosorbent (RAST) with 129 sera, using media extracts on the discs . Purified mite body extract (PMB) contained less ag 42 and more ag Y and ag 23 than whole mite culture extract ( WMC ), whereas an acetone-extracted mite excreta preparation (AML) contained 5 times more ag 42, but was devoid of ag Y and ag 23 . Ag X was present in all preparations . The RAST-inhibitory potency of PMB was best correlated with the content of ag X . Preparations with properties similar to WMC and PMB were judged as suitable for clinical application.

Poult Sci, 1984 May, 63(5), 1020 - 6
Effect of fractions of fish meal and hepatic lipid deposition in estrogenized chicks; Mendonca CX Jr et al.; Three experiments were conducted for the purpose of determining the effect of feeding isoenergetic diets with and without fish meal or its fractions on hepatic lipid deposition in estrogenized chicks . The chicks were fasted for 48 hr before being fed the experimental diets and were injected with estradiol three times during the experiment . After 4 days, hepatic lipid content was determined . In the first experiment, the quantity of hepatic lipid deposited per unit of liver and per 100 g body weight was significantly less for chicks fed 10% fish meal and an ethanol extract of fish meal equivalent to 10% than for chicks fed an unsupplemented corn-soy diet . In a second experiment a significant reduction in hepatic lipids deposited per 100 g body weight was observed with 10% fish meal but only a numerical reduction with ethanol extracts of fish meal equivalent to 10 or 20% and the ash of ethanol extract equivalent to 20% . In a third experiment a highly significant reduction in hepatic lipid deposition was observed in birds refed a diet containing fish meal, alfalfa, and torula yeast but feeding this before the starvation-refeeding period did not affect liver lipids . These results show that fish meal contains hepatic lipid lowering activity in refed estrogenized chicks and suggest that the activity may be extracted with ethanol.

J Virol, 1984 May, 50(2), 287 - 92
Influenza virus-induced immune complexes suppress alveolar macrophage phagocytosis; Astry CL et al.; Immune complexes in the lungs are capable of inducing adverse responses . Herein we have detailed the formation of immune complexes in the lungs of influenza virus-infected mice and examined their effect on alveolar macrophage defenses . On days 3, 7, 10, 15, and 30 after aerosol infection with influenza A/PR8/34 virus, the acellular pulmonary lavage fluid was tested for viral antigen, specific viral antibody, and immune complexes by immunoassays . Whereas peak viral antigen (day 3) diminished to undetectable levels by day 10, specific viral antibody remained at a low concentration until day 10, after which it rapidly increased . Immune complex concentrations increased through day 7, peaked at day 10, and gradually returned to the control level by day 30 . These data demonstrate that immune complexes of detectable size are induced by influenza virus infection during the interface between antigen excess and antibody excess conditions . Since alveolar macrophages are the pivotal phagocytic defense cells in the lung, the ability of normal alveolar macrophages to ingest opsonized erythrocytes was quantitated in the presence of immune complexes from lavage fluid . Immune complexes from day 10 virus-infected lungs caused a dose-dependent suppression of antibody-mediated phagocytosis to 30% of control values . In contrast, although these immune complexes also markedly decreased the phagocytosis of antibody-coated yeast cells, they did not significantly impair the antibody-independent ingestion of unopsonized yeast cells by macrophages . the suppressive effects of immune complexes on alveolar macrophages may, in part, explain the phagocytic dysfunction that occurs 7 to 10 days after influenza virus pneumonia.

Biofizika, 1984 May-Jun, 29(3), 419 - 23
{Changes in the electrical and visco-elastic properties of bilayer lipid membranes during interaction with proteins and lipoproteins}; Shimane Ch et al.; A method for simultaneous registration of planar bilayer lipid membrane (BLM) DC conductance G, capacitance C, surface potential difference delta phi and transversal elasticity module E is developed . C, delta phi and E are proportional to the amplitude of the first, second and third harmonics of capacitance current respectively . A comparative study of the interaction of BLM with very low density lipoproteins (VLDL), influenza virus matrix protein (M-protein) and yeast invertase was carried out . The kinetics of delta phi, E and G changes at different concentrations of VLDL, and dependence of delta phi and G on M-protein and invertase concentration was investigated . It is shown for VLDL invertase and M-protein that the changes in delta phi and E occur before the change in G . The method used permits to study peculiarities of individual stages of interaction between charge particles, supramolecular structures and lipid membranes.

J Clin Immunol, 1984 May, 4(3), 246 - 9
Hyaluronic acid enhances phagocytosis of human monocytes in vitro; Ahlgren T et al.; Pretreatment of human blood monocytes with hyaluronic acid for 0.5 hr enhanced significantly their ingestion in vitro of yeast particles opsonized with complement . The enhancement of the phagocytic capacity was dose dependent and maximal in the range of 25-50 micrograms/ml.

J Pediatr, 1984 May, 104(5), 706 - 9
Differentiation of lymphoma from histoplasmosis in children with mediastinal masses; Gaebler JW et al.; The differentiation of mediastinal masses caused by lymphoma from those caused by histoplasmosis may require thoracotomy . We reviewed the medical records of 37 children undergoing initial evaluation for anterior or middle mediastinal masses . Sixteen had biopsy-proved lymphoma, and 21 had histoplasmosis; seven with histoplasmosis underwent thoracotomy . Age, sex, fever, weight loss, duration of illness, anemia, erythrocyte sedimentation rate, nonspecific reactants, and lung infiltrates and calcifications were similar in both groups . Masses were in the middle mediastinum in all patients with histoplasmosis and in 69% with lymphoma . Masses were in the anterior mediastinum in one of 21 (5%) with histoplasmosis and 13 of 16 (81%) with lymphoma . Among patients with lymphoma, histoplasmal complement fixation antibody titers were less than 1:8 in 14 of 15 (93%); a single patient had a titer of 1:16 . The CF titers were greater than or equal to 1:32 in 14 of 21 (67%) with histoplasmosis . In children with middle mediastinal masses, a histoplasmal CF yeast or mycelial titer greater than or equal to 1:32 is strongly suggestive of acute histoplasmosis and biopsy is not required . Children not fulfilling these criteria should undergo diagnostic biopsy.

Mol Biochem Parasitol, 1984 May, 12(1), 37 - 48
Selective acquisition of plasma proteins by Trichomonas vaginalis and human lipoproteins as a growth requirement for this species; Peterson KM et al.; Trichomonas vaginalis avidly bound numerous host macromolecules which were not removed by repeated washing in phosphate buffered saline . The use of radioiodinated Cohn plasma fractions in binding studies allowed the identification of plasminogen, fibrinogen, immunoglobulin G, lipoproteins A and B, transferrin, alpha 1-antitrypsin, and albumin on intact organisms . The binding of immunoglobulin G, albumin, transferrin, and lipoproteins to intact, motile trichomonads was further demonstrated using 125I-labeled plasma that was chromatographically depleted of these proteins . Kinetic studies indicated that 125I-labeled lipoproteins bind to T . vaginalis in a receptor-ligand-like manner . The surface localization and uptake of bound lipoproteins was shown by treatment of intact organisms with pronase at various times after incubation with lipoproteins . Purified lipoproteins could be substituted for plasma or serum as a growth supplement in a complex medium of trypticase/yeast extract/maltose and supported growth and multiplication rates equal to those in the same medium with plasma.

Antimicrob Agents Chemother, 1984 May, 25(5), 560 - 2
In vitro activities of amphotericin B in combination with four antifungal agents and rifampin against Aspergillus spp; Hughes CE et al.; Strains of Aspergillus fumigatus, Aspergillus flavus, and Aspergillus niger were tested for in vitro susceptibility with a microtiter plate system in buffered yeast-nitrogen base and in buffered minimal essential medium . Isolates were tested against amphotericin B, flucytosine, rifampin, ketoconazole, ICI 153,066, and Bay n 7133 and against combinations of amphotericin B with each of the other five drugs . Combinations of amphotericin B and rifampin were the most active against all three species of Aspergillus . Flucytosine combined with amphotericin B produced little or no reduction of the MICs at which 90% of the strains were inhibited compared with amphotericin B alone . With one exception, the addition of ketoconazole, ICI 153,066, or Bay n 7133 to amphotericin B did not consistently alter the MICs . The addition of ICI 153,066 markedly increased the MICs of amphotericin B against the A . flavus isolates in both media . When the azoles were tested alone, Bay n 7133 was the most active against A . fumigatus, but was two- to fivefold less active against A . flavus . Ketoconazole was the most active azole against A . flavus.

J Biochem (Tokyo), 1984 May, 95(5), 1355 - 66
Cross-reactivity of contact site A to antibody produced against a stage-specific antigen from a phenol/water extract of Dictyostelium discoideum; Hirano T et al.; The stage-specific antigen, gp68 (Hirano, T., Yamada, H., & Miyazaki, T . (1983) Biochim . Biophys . Acta 742, 224-234), was purified from a phenol/water extract of aggregation-competent cells of Dictyostelium discoideum by preparative polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS) . Anti-gp68 was produced against purified gp68 which was determined to be homogeneous by silver staining on analysis by SDS-polyacrylamide gel electrophoresis . The cross-reactivity of anti-gp68 against cellular antigens was estimated by immuno-affinity chromatography on anti-gp68 immunoglobulin G (IgG)/Sepharose . When the whole cell lysate was applied to this affinity column, three major proteins, with molecular weights of 80,000, 68,000, and 56,000, were obtained in the absorbed fraction . When the butanol extract, which was enriched in contact site A, an adhesion mediating glycoprotein, was applied to the same column, two major proteins with molecular weights of 80,000 (corresponding to contact site A) and 56,000 were obtained in the absorbed fraction but, however, gp68 was negligible . Reversely, when the phenol/water extract was applied to anti-contact site A-IgG/Sepharose, only gp68 was obtained in the absobed fraction . Moreover, contact site A was seen to compete with {3H}mannose-labeled gp68 in a competition radioimmunoassay using anti-gp68 serum . The effect of Pronase or exo-alpha-mannosidase digestion on the antigenic activity of gp68 was examined by radioimmunoassaying . The results indicated that the alpha-mannosyl residue of the non-reducing terminal in the carbohydrate moiety of gp68 was a major immunodeterminant . However, the polypeptide chain did not participate in the antigenic reactivity against anti-gp68 . Both anti-gp68 and anti-contact site A agglutinated heat killed-yeast cells . Also, both anti-sera inhibited EDTA-stable cell adhesion of aggregation-competent cells in the presence of Fab from goat anti-rabbit IgG . These results indicate that gp68 and contact site A have a common antigenic determinant against anti-gp68, and that the target antigen of anti-gp68 was somehow involved in cell adhesion.

Vopr Pitan, 1984 May-Jun, (3), 41 - 4
{Comparative biological value of protein hydrolysates and their fortified forms}; Safronova AM et al.; Experiments on Wistar male rats were made to study the biological value (with the use of the "growth" method) of basic and protein- and crystalline amino acids-enriched protein hydrolysates . Autolysine and protein hydrolysates obtained from formed elements of the blood of slaughtered animals (the degree of degradation 30 and 70%) and a mixture of crystalline amino acids composed according to the FAO/WHO scale were studied . Casein was used as control . The differences in the biological value of basic and protein hydrolysates-enriched preparations point to the necessity of correcting basic hydrolysates.

Mycopathologia, 1984 Apr 30, 86(1), 29 - 33
Effects of caffeine, cyclic 3', 5' adenosine monophosphate and cyclic 3', 5' guanosine monophosphate in the development of the mycelial form of Sporothrix schenckii; Rodriguez-Del Valle N et al.; The kinetics of the development of the mycelial form of Sporothrix schenckii from yeast cells and conidia in a minimal basal medium with glucose at pH 4.0 and 25 degrees C were established . Germ tube formation was used as the index of germination for both yeast cells and conidia . Yeast cells were first observed to develop germ tubes after 3 h of incubation, reaching 92 +/- 5%, after 12 h of incubation . Germ tubes were first detected in conidia after 9 h of incubation, and 12 h after inoculation 92 +/- 6% of the conidia had germ tubes . After 24 h of incubation, fully developed, sporulating mycelia were observed from both yeast cells and conidia . A delay in germ tube formation from yeast cells was observed when But2cAMP (10 mM) and But2cGMP (10 mM) were added to the medium . Also the addition of caffeine, a cyclic nucleotide phosphodiesterase inhibitor, inhibited the yeast to mycelial transition . Conidial germination into the mycelial form was also inhibited when cAMP, But2cAMP and caffeine were added to the medium . These results suggest the possible involvement of cyclic nucleotides in the control of dimorphism in S . schenckii.

Biochem Biophys Res Commun, 1984 Apr 16, 120(1), 124 - 30
Interaction of reduced and oxidized cytochrome c with the mitochondrial cytochrome c oxidase and bc1-complex; Bill K et al.; Making use of a hetero-bifunctional reagent (succinimidyl 4-(p-male-imidophenyl)butyrate, SMPB), yeast cytochrome c was linked through a thioether bond to the maleimide group whereas the active N-hydroxy-succinimide ester site of the SMPB was used for the reaction with the primary amino groups of Affi-gel 102 . The capacity and stability (also to reducing agents) of the column were greatly improved relative to previous systems . This new gel allowed the study of the interactions of cytochrome c oxidase and reductase with reduced and oxidized cytochrome c . For cytochrome c oxidase a significant difference in the interaction with ferri- and ferro-cytochrome c was observed but no such a difference was seen in the case of cytochrome c reductase.

Mycopathologia, 1984 Apr 15, 85(3), 167 - 70
Vaginal candidosis . Opportunistic factors and clinical correlation in 600 patients; Lopez-Martinez R et al.; Vaginal exudates were taken from 600 new patients of the gyneco -obstetrics outpatient clinic . Candida was isolated from 261 patients, 134 (22.3%) of which had this yeast as a component of the normal flora, and in 127 (21.2%) it was considered as a pathogen . The most frequent symptoms in the last group were vaginal discharge, erythema and pruritus . Pregnancy was the most frequent opportunistic factor, followed by the association of pregnancy and malnutrition, and anemia . Vaginal candidosis was more frequent in patients of the medium socio-economical stratum . The species of Candida isolated were C . albicans (67.7%), C . tropicalis (18.8%), C . stellatoidea (8.7%), C . pseudotropicalis (2.4%), C . parakrusei (1.6%) and C . guillermondi (0.8%).

Nucleic Acids Res, 1984 Apr 11, 12(7), 3405 - 23
Mapping of psoralen cross-linked nucleotides in RNA; Garrett-Wheeler E et al.; A method is described for using the cross-linking reagent 4'-(hydroxy-methyl)-4,5',8-trimethylpsoralen (HMT) to map base paired regions and higher-order structure within RNA molecules . Applying this method to yeast tRNAPhe, we have specifically identified cross-links within the acceptor stem between U6 X U68, in the D-stem between C11 X C25, and in the T psi-stem between U50 X C63 and U52 X C63 . We have also identified a unique cross-link between U8 X C48 which are trans pyrimidines in the core region due to tertiary interactions between U8:A14 and C48:G15 . The precise point of cross-linking was deduced in every case by using purine-specific U2 ribonuclease along with cytidine-specific CL3 ribonuclease which will anomalously cleave after photoreversed pyrimidines . The ability to map the precise point of cross-linking should prove invaluable in identifying nucleotides in close proximity within the tertiary structure of other RNA molecules.

Arch Biochem Biophys, 1984 Apr, 230(1), 285 - 93
Orotate phosphoribosyltransferase and orotidylate decarboxylase from Crithidia luciliae: subcellular location of the enzymes and a study of substrate channeling; Pragobpol S et al.; Orotate phosphoribosyltransferase (OPRTase) and orotidylate decarboxylase (ODCase) have been found to be particulate in the kinetoplastid protozoan, Crithidia luciliae . Sucrose density centrifugation indicated that these two enzymes are associated with the glycosome, a microbody which appears to be unique to the Kinetoplastida and which contains many of the glycolytic enzymes . The particulate location of OPRTase and ODCase was considered to be favorable for channeling of orotidine-5'-monophosphate (OMP), the product of the first enzyme and substrate for the second . The degree of channeling was determined by double radioactively labeled experiments designed to determine the relative efficiency of endogenous and exogenous OMP as substrates of ODCase . The efficiency of channeling was high, with an approximate 50-fold preference for endogenous OMP . By comparison, the degree of channeling for the yeast enzymes, which are soluble and unassociated, was less than 2-fold . The OPRTase-ODCase enzyme complex was solubilized using Triton X-100 in the presence of dimethyl sulfoxide, glycerol, and phosphoribosyldiphosphate . The percentage recovery of the overall enzyme activity was approximately 20% . The degree of channeling was reduced by approximately 10-fold for the solubilized complex . The Km for OMP changed from 7.5 (+/- 1.8) to 1.6 (+/- 0.3) microM in the ODCase reaction . There was no alteration in the Km for orotate in the OPRTase reaction.

Biochem J, 1984 Apr 1, 219(1), 341 - 4
Tertiary structure of tRNAPhe . A possible correlation between the structural functional unit of this tRNA and its exonic sequence; Malathi R et al.; It has been shown recently {Go (1981) Nature (London) 291, 90-92; Blake (1983) Trends Biochem Sci . 8, 11-13} that the exonic regions of the genes of proteins haemoglobin, lysozyme and immunoglobin correspond closely to the compactly folded structural units . Despite the absence of classical domain structures in tRNA compared with those found in several proteins, close inspection of certain features in the distance maps obtained for yeast tRNAPhe using the conformationally equivalent heminucleotide scheme reveals that a similar situation might also be present in ribonucleic acids such as tRNA species and the exonic sequences of their genes . Also it seems possible that certain segments of yeast tRNAPhe may be characterized as possessing a domain-like character, and this seems to provide stereochemical support for possible conservation of L-shape structure for tRNA species lacking the entire dihydrouridine arm such as those found in mitochondria.

Proc Natl Sci Counc Repub China B, 1984 Apr, 8(2), 182 - 6
Biosynthetic studies of amphotericins, candicidin and nystatin by means of mutation; Liu YT; A novel mutant strain designated as Streptomyces nodosus, NDMC-034 was selected from the mutation of Streptomyces nodosus, IMRU 3694 . This mutant was characterized by delayed sporulation, the incapability of producing the melanoid pigment either in the production medium or on YD agar containing 1% yeast extract and 1% dextrose, and the increased yield of amphotericin A as compared to its parent strain . In addition, the biosynthesis of amphotericin A by this mutant seems to be sensitive to temperature . When the incubation temperature was raised to 32 degrees C, the yield of amphotericin A was dramatically decreased; in contrast, the yield of amphotericin B was greatly increased as compared to that at 28 degrees C . Another unique mutant strain designated as Streptomyces griseus, YT-1 was selected from the mutation of Streptomyces griseus, IMRU 3570 . It was characterized by the incapability of synthesizing candicidin and the delayed sporulation . All the mutants were auxanographed and characterized by the requirement for growth factor.

Gen Comp Endocrinol, 1984 Apr, 54(1), 76 - 84
Ecdysteroids during the third larval instar in 1(3)ecd-1ts, a temperature-sensitive mutant of Drosophila melanogaster; Berreur P et al.; The temperature-sensitive 1(3)ecd-1ts mutation (A . Garen, L . Kauvar, and J.A . Lepesant (1977) . Proc . Natl . Acad . Sci USA 74, 5099-5103.) has been used in several laboratories to obtain Drosophila larvae deprived of moulting hormone . The development of mutants and controls during the third larval instar at permissive (20 degrees C) and restrictive temperatures (29 degrees C) was compared . Pupariation was inhibited when larvae were shifted to the restrictive temperature immediately at the second moult . The permanent larvae obtained remained active, did not leave the food, and reached a maximum weight superior to the weight of controls . Ecdysteroids were studied during the third larval instar by HPLC analysis and radioimmunoassays . A careful synchronization of the larvae at the second moult enabled the confirmation that at least one ecdysteroid peak occurs during the third larval instar, prior to the wandering stage in controls (20 or 29 degrees C) . Ecdysone was then the predominant moulting hormone, whereas 20-hydroxyecdysone was the main ecdysteroid at the time of pupariation . Low levels of ecdysteroid were measured in mutant larvae shifted to 29 degrees C immediately at the second moult but larvae completely deprived of immunoreactive material were never observed . Nearly normal levels of ecdysteroids appeared at 27.5 degrees C . Feeding ecd-1 larvae maintained at restrictive temperature on 20-hydroxyecdysone-yeast mixture for 16 hr triggered abortive pupariation . Ecdysteroid levels were measured after the return of the larvae to the standard medium; normal levels were restored 24 hr later . The mutant ecd-1 appears to present interesting opportunities for the detailed study of the hormonal induction of a developmental process during the third larval instar.(ABSTRACT TRUNCATED AT 250 WORDS)

Pathology, 1984 Apr, 16(2), 179 - 83
Dermatophytes identified at the Australian National Reference Laboratory in Medical Mycology 1966-1982; Muir DB et al.; Over a 17 yr period from 1966 to 1982, 4354 dermatophytes were identified at the Australian National Reference Laboratory in Medical Mycology . The most frequently identified species was Trichophyton rubrum, accounting for 35.3% of identifications, followed by Trichophyton mentagrophytes (26.5%), Trichophyton tonsurans (12.8%), Epidermophyton floccosum (10.7%) and Microsporum canis (8.4%) . Specimens taken from the feet were the most common source of the Trichophyton rubrum isolates, followed by specimens from the groin . The highest incidence of T . rubrum and other Trichophyton infections was found in males in the age group 21-30 yr . Microsporum species were most commonly isolated from children aged 10 yr or less . In 1.5% of specimens, more than one fungal species was isolated . In most of these instances a yeast (predominantly Candida species) was found in addition to a Trichophyton species.

Anal Biochem, 1984 Apr, 138(1), 164 - 80
Optimization of in vitro protein synthesis by isolated mouse adrenal mitochondria; Mills NC et al.; The requirements for in vitro mitochondrial protein synthesis have been studied using isolated mitochondria from cultured adrenal Y-1 tumor cells from mice . By reducing the reaction volume to 50 microliter we were able to assay in replicate the requirements for various reaction components using trichloroacetic acid (TCA)-precipitable counts for a quantitative evaluation with time of incubation . Sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis followed by autoradiography was also used for a qualitative and quantitative evaluation of the translation products . With the optimized system, 1 to 3% of added {35S}methionine was incorporated . The products of mitochondrial protein synthesis range from 70,000 to 5000 molecular weight . Major autoradiographic bands were observed at 38,000, 31,000, 23,000, 20,000, and 5600 molecular weight as separated on 10 to 20% gradient SDS-polyacrylamide gels; however, 20 to 30 protein products of various molecular weights were discernible . Mitochondrial concentrations of 0.8 to 1.4 mg/ml of incubation gave the better incorporation of {35S}methionine per milligram of protein . Total {35S}methionine incorporated into mitochondrial protein was greatest at 25 degrees C after 90 min . Chloramphenicol at 10 micrograms/ml inhibited mitochondrial protein synthesis by more than 50% and at 100 micrograms/ml inhibited incorporation by more than 95% . Cycloheximide had no effect on incorporation at less than 1.0 mg/ml . Magnesium and ATP in a molar ratio of one to one at 5 mM gave optimal incorporation . Other energy generating systems using oxidative phosphorylation to supply ATP for protein synthesis were not as effective as ATP and 5 mM phosphoenol pyruvate, 20 micrograms/ml pyruvate kinase and 5 mM a-ketoglutarate . In contrast to in vitro yeast mitochondrial protein synthesis, no enhancement of in vitro adrenal cell mitochondrial protein synthesis was found with GTP or its analogs . The buffers N,N-bis(2-hydroxyethyl)glycine, N-(tris(hydroxymethyl)methyl)glycine, and N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid were superior to Tris-HCl for mitochondrial protein synthesis . Optimal pH for {35S}methionine incorporation into mitochondrial proteins was pH 7.0 to 7.6 . Potassium at 50 to 90 mM gave the best incorporation of {35S}methionine, and the higher molecular weight products of translation were enhanced at these concentrations . Sodium at 10 to 40 mM had no effect; however, 100 mM sodium inhibited label incorporation by 30% . Calcium at 100 microM inhibited mitochondrial protein synthesis by approximately 50%, and at 1.0 mM little if any incorporation occurred.(ABSTRACT TRUNCATED AT 400 WORDS)

Aust J Exp Biol Med Sci, 1984 Apr, 62 ( Pt 2), 117 - 23
Immunogenicity of RNA-protein fraction isolated from axenic Entamoeba histolytica (NIH:200); Sharma GL et al.; The immunogenicity of Entamoeba histolytica RNA (EhRNA) was studied in guinea-pigs . Animals immunized with a single dose of EhRNA and Freund's complete adjuvant (FCA) showed a high level of cell-mediated immune response (CMIR) as assessed by percent leucocyte migration inhibition (74.8 +/- 2.9) after 1 week of immunization . This response faded away gradually and by week 5 after immunization the percent leucocyte migration inhibition was 27.1 +/- 8.27, which is quite close to control values . The immunogenic activity of EhRNA was found to be specific, since yeast RNA failed to elicit immunological responses, and was RNase-sensitive . The CMIR could be maintained at its high level by giving boosters or multiple doses . Anti-EhRNA antibodies (haemagglutinins or precipitins) could not be demonstrated at any stage.

J Clin Invest, 1984 Apr, 73(4), 1062 - 71
Functional maturation of membrane potential changes and superoxide-producing capacity during differentiation of human granulocytes; Kitagawa S et al.; The alterations of stimulus-induced membrane potential changes, superoxide (O2-)-producing capacity and phagocytic activity during differentiation of human granulocytes were investigated in the human leukemia cell lines HL-60 and KG-1 differentiating in vitro and in human leukemic granulocytes obtained from chronic myelogenous leukemia patients . HL-60 cells incubated with dimethyl sulfoxide or with retinoic acid showed progressively increasing O2- production as well as membrane potential changes (depolarization) on contact with phorbol myristate acetate or the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine, with a concomitant increase in the proportion of mature cells of the granulocytic type . Phagocytosis of latex particles, yeast, and oil droplets appeared 24 h after incubation with dimethyl sulfoxide and anteceded the increment of O2- production and membrane potential changes, both of which appeared concomitantly 3 d after incubation with dimethyl sulfoxide . Similar findings were observed when immature and mature granulocytes obtained from chronic myelogenous leukemia patients were stimulated by phorbol ester, the chemotactic peptide, or calcium ionophore A23187, and the amount of O2- production was parallel to the magnitude of membrane potential changes . HL-60 and KG-1 cells incubated for 1-6 d with phorbol myristate acetate showed neither O2- production nor membrane potential changes on contact with phorbol ester, chemotactic peptide, or A23187, although such cells resembled macrophages morphologically, and their phagocytic activity was significantly increased . O2- production and membrane potential changes in normal granulocytes induced by phorbol ester, chemotactic peptide and A23187 were inhibited by 2-deoxyglucose . These findings indicate that the O2--producing system and the system provoking membrane potential changes may develop concomitantly as human granulocytes mature and differentiate, and that the development of these systems and of phagocytic activity may be independently regulated.

Biochem Biophys Res Commun, 1984 Mar 30, 119(3), 1008 - 14
Phosphorylation of glucose analog 3-O-methyl-D-glucose by rat heart; Gatley SJ et al.; When isolated rat hearts were perfused with medium containing 3-O-methyl-D-glucose (3MG) a compound with the chromatographic behavior expected for 3MG-6-phosphate was formed . This assignment is supported by the action of alkaline phosphatase and by production of an identical peak on incubation of 3MG with yeast hexokinase in a buffer containing adenosine triphosphate and Mg++ ions . The clearance of labeled 3MG from isolated perfused rat hearts was biphasic and eventually total . Since sugar phosphates do not cross cellular membranes, this behavior is consistent with both formation and dephosphorylation of 3MG-6-phosphate, with dephosphorylation rate-limiting the slow phase of the clearance . Phosphatase activity was also indicated by a decrease in 3MG-6-phosphate when hearts were not chilled and homogenized immediately after perfusion . The common assumption that 3MG is not metabolized after transport into cells is shown by these experiments to be erroneous.

Science, 1984 Mar 16, 223(4641), 1189 - 91
Phenylalanine transfer RNA: molecular dynamics simulation; Harvey SC et al.; Yeast phenylalanine transfer RNA was subjected to a 12-picosecond molecular dynamics simulation . The principal features of the x-ray crystallographic analysis are reproduced, and the amplitudes of atomic displacements appear to be determined by the degree of exposure of the atoms . An analysis of the hydrogen bonds shows a correlation between the average length of a bond and the fluctuation in that length and reveals a rocking motion of bases in Watson-Crick guanine X cytosine base pairs . The in-plane motions of the bases are generally of larger amplitude than the out-of-plane motions, and there are correlations in the motions of adjacent bases.

Eur J Biochem, 1984 Mar 15, 139(3), 481 - 7
Specific changes in lactate levels, lactate dehydrogenase patterns and cytochrome b559 in Dictyostelium discoideum caused by queuine; Schachner E et al.; Higher eukaryotes contain tRNA transglycosylases that incorporate the guanine derivative queuine from the nutritional environment into specific tRNAs by exchange with guanine at position 34 . Alterations in the queuosine content of specific tRNAs are suggested to be involved in regulatory mechanisms of major routes of metabolism during differentiation . Dictyostelium discoideum has been applied as a model to investigate the function of queuine or queuine-containing tRNAs . Axenic strains are supplied with queuine by peptone, but they grow equally well in a defined queuine-free medium . Queuine-lacking amoebae, starved in suspension culture for 24 h, lose their ability to differentiate into stalk cells and spores, whereas amoebae sufficiently supplied with queuine will overcome this metabolic stress and undergo further development when plated on agar . The results presented here show that D(-)-lactate occurs in the slime mould in millimolar amounts and that its level is remarkably decreased in queuine-lacking cells after 24 h of starvation in suspension culture . On isoelectric-focusing polyacrylamide gels, nine different forms of NAD-dependent D(-)-lactate dehydrogenase can be separated from extracts of vegetative cells, and six forms from extracts of the starved cells . Under queuine limitation, one form is missing in the starved cells . Low amounts of L(+)-lactate are usually found in vegetative amoebae but significantly less in queuine-lacking cells . Five forms of NAD-dependent L(+)-lactate dehydrogenase are detectable in extracts from vegetative, queuine-treated cells, and slight alterations occur in queuine-deficient amoebae . In the starved cells only one form of L(+)-lactate dehydrogenase is found, irrespective of the supply of queuine to the cells . A cytochrome of type b with an absorption maximum at 559 nm accumulates during starvation only in queuine-lacking cells; it might be a component of an NAD-independent lactic acid oxidoreductase as is cytochrome b 557 in yeast and be responsible for the reduced level of lactate in cells lacking queuine in tRNA.

Acta Neurol Scand, 1984 Mar, 69(3), 147 - 53
Changes in response to metrazole during fever in juvenile rats; a new model for febrile convulsions?
MacKintosh DA, Baird-Lambert J, Buchanan N.
Previous models for febrile convulsions have used environmentally induced hyperthermia as the stimulus to induce convulsions . Changes in response to metrazole during yeast-induced fever in juvenile rats are reported here . Animals were more susceptible to metrazole during the rising phase of fever but showed some resistance to its convulsant effects once the fever was established and following defervescence . It is suggested that this may form the basis of a physiologically more appropriate model for the study of the pathogenesis of febrile convulsions.

Poult Sci, 1984 Mar, 63(3), 524 - 31
Diet composition, environmental temperature, and exogenous estradiol effects on hepatic lipid deposition in growing chicks; Takahashi K et al.; The interrelationships among dietary composition, environmental temperature, and level of estradiol (E2) administration on hepatic lipid deposition in chicks were studied . Two levels of E2 were injected at three intervals over a 4-day period, in 3-week-old male Leghorn chicks fed either a corn-soybean meal (CS) diet or a diet containing fish meal (FM) after 2 days fasting . The chicks were subjected to temperature ranges of 15 to 24 C (low) or 24 to 35 C (high) at 2 weeks of age . The E2 (in silastic tubes) was also implanted subcutaneously in 3-week-old broiler chicks fed either the CS diet or a diet containing fish meal, alfalfa meal, and torula yeast (FAY) from day of age to 6 weeks . They were subjected to the high and low temperature ranges at 3 weeks . Liver lipid deposition markedly increased with E2 administration among chicks fed all diets within both temperature ranges . Liver lipid was significantly greater at 24 to 35 C than at 15 to 24 C among estrogenized chicks . Feeding the FM or the FAY diet decreased hepatic lipid accumulation as compared to feeding the CS diet, but the ameliorative effect of the FM diet on hepatic lipid deposition was not observed at the low temperature or at the lower level of implantation . It was concluded that a range for estrogen administration exists wherein dietary effects are expressed . These data indicate that environmental temperature, dietary composition, estrogen level, and their interactions influence hepatic lipid deposition and also suggest that high temperature augments liver response to estrogen.

J Nutr, 1984 Mar, 114(3), 591 - 7
Responses in calcium and phosphorus metabolism and hepatic lipid deposition among estrogenized chicks fed various dietary ingredients; Bolden SL et al.; The purpose of this study was to determine whether diet composition would influence calcium and phosphorus metabolism in chicks administered estrogen . At 1 day of age, broiler chicks were fed either a corn-soybean meal diet (CS), or an isoenergetic and isonitrogenous diet containing 5% fish meal, 5% alfalfa meal and 10% torula yeast (FAY) . At 21 days equivalent numbers were implanted with one of two lengths of Silastic tubing containing estradiol dipropionate, while the remaining birds served as nonimplanted controls . Significant increases were observed in liver weight, liver lipid, plasma total calcium and inorganic phosphate in chicks that were implanted, while concomitant declines were seen in body weight . Implanted chicks fed the CS diet had significantly higher liver weight, liver lipid, plasma phosphorus and plasma calcium and lower tibial bone ash than those fed the FAY diet . Furthermore, liver lipid values were very highly correlated with plasma phosphorus and calcium . In an identical study with slower growing White Leghorn chicks, the same trends were observed but were less well defined . These data show that the inclusion of certain ingredients into corn-soybean diets balanced for the major nutrients affects the response of chicks to estrogenization . Liver lipid deposition, calcium and phosphorus metabolism are all subject to diet and estrogen interactions.

J Bacteriol, 1984 Mar, 157(3), 899 - 908
Characterization of Saccharomycopsis lipolytica mutants that express temperature-sensitive synthesis of isocitrate lyase; Matsuoka M et al.; Four mutants specifically deficient in the activity of isocitrate lyase were independently isolated in the alkane yeast Saccharomycopsis lipolytica . Genetic analysis by means of protoplast fusion and mitotic haploidization revealed that the mutations were recessive and non-complementary at a single genetic locus, icl . icl is a structural gene for isocitrate lyase, because some revertants from icl-1 and icl-3 mutants produced thermolabile isocitrate lyase in comparison with the wild-type enzyme, and also because the gene dosage effect was observed on the specific activity of isocitrate lyase in icl+/icl-1 and icl+/icl-3 heterozygotes . The icl-3 mutation also gave rise to temperature-sensitive revertants that could grow on acetate at 23 degrees C but not at 33 degrees C, exhibiting temperature-sensitive synthesis as well as thermostable activity of isocitrate lyase . Studies on purified isocitrate lyase showed that this enzyme is tetrameric and that the enzyme synthesized at 23 degrees C by a temperature-sensitive synthesis mutant was indistinguishable from the wild-type enzyme with respect to the subunit molecular weight (59,000), the isoelectric pH (5.3), the thermostability, and the Km value for threo-Ds-isocitrate (0.2 mM) . When induced by acetate at 33 degrees C, the temperature-sensitive synthesis mutant did not express isocitrate lyase activity but did synthesize polypeptides whose electrophoretic mobilities were equal to that of the purified mutant enzyme . Hence, the temperature-sensitive mutation assumed in the structural gene for isocitrate lyase might have prevented the maturation of the polypeptide chains synthesized at the restrictive temperature.

Exp Cell Res, 1984 Mar, 151(1), 247 - 51
Visualization of reactive oxygen species formation by phagocytizing macrophages; Peskin AV et al.; Activated peritoneal macrophages exhibiting phagocytosing capacity produced an electron-dense precipitate of formazan in contact sites of macrophage plasmalemma and phagocytosed yeast cells . No production of formazan occurred, when non-opsonized latex particles were ingested by macrophages . Formazan precipitation could be prevented by anaerobiosis but not by addition of cyanide.

Arch Fr Pediatr, 1984 Mar, 41(3), 193 - 5
{Purulent pleurisy, esophagopleural fistula and presence of Torulopsis glabrata in the pleura}; Goldszmidt D et al.; The authors report the simultaneous discovery of a septic pleural effusion and an esophagopleural fistula in a 4 year-old patient . The yeast Torulopsis glabrata found in the pleural effusion does not seem to behave as a pathogen but more likely as a witness of a fistula between the gastrointestinal tract and the pleura.

Exp Cell Res, 1984 Mar, 151(1), 183 - 93
Indirect immunofluorescence of microtubules in Dictyostelium discoideum . A study with polyclonal and monoclonal antibodies to tubulins; Roos UP et al.; Amebae of D . discoideum on coverslips were fixed in situ with glutaraldehyde and permeabilized with Triton X-100 . Of six antibodies tested, only a monoclonal antibody to yeast tubulin consistently gave bright fluorescence . Counterstaining with DAPI facilitated the identification of interphase and mitotic stages . Most microtubules (MTs) in interphase amebae emanated from a nucleus-associated centre that had a non-fluorescent core . Amebae in early stages of mitosis lacked cytoplasmic MTs almost entirely . The nascent spindle in prophase appeared as a brightly fluorescent dot, whereas the prometaphase spindle was a short rod . Spindles in metaphase and anaphase nuclei were more elongate, some consisting of several fluorescent lines . Astral MTs were prominent on spindles in anaphase and telophase . Asters are obviously converted to the interphase complex of MTs in post-mitotic cells, while the shaft-like remnant of the central spindle disappears . The cyclical changes in the MT system related to cell division resemble those observed in higher eukaryotes and probably reflect changes in the locomotory behavior of the amebae rather than changes in cell shape.

Arch Intern Med, 1984 Mar, 144(3), 549 - 51
Efficacy of ketoconazole v nystatin in prevention of fungal infections in neutropenic patients; Jones PG et al.; A prospective randomized study was undertaken in neutropenic patients to evaluate the efficacy of prophylactic ketoconazole v nystatin in reducing yeast infections . Eighteen patients received 500,000 units of nystatin suspension four times daily, and 18 patients received 200 mg of ketoconazole daily . The nystatin group experienced nine local yeast infections (four thrush, three esophagitis, and two vaginitis); three patients receiving ketoconazole had thrush . No cases of disseminated candidiasis occurred in either group . Ketoconazole was better tolerated than nystatin and neither drug caused toxic effects . In addition to being nontoxic and better tolerated, ketoconazole appeared to be slightly more effective than nystatin in reducing locally severe yeast infections.

J Cell Sci, 1984 Mar, 66, 265 - 75
The mitotic spindle of Chinese hamster ovary cells isolated in taxol-containing medium; Kuriyama R et al.; Mitotic spindles from Chinese hamster ovary (CHO) cells were isolated and purified by a one-step procedure in an isolation medium containing the microtubule-stabilizing drug, taxol . Released mitotic spindles were examined by phase-contrast, polarizing and differential-interference microscopy . They were also stained with monoclonal antibody raised against yeast tubulin and examined by epifluorescence microscopy . The spindles were free from visible cytoplasmic contaminants and the chromosomes were generally lost from the preparations . Electron microscopy showed that microtubules were the dominant structural component and sodium dodecyl sulphate/gel electrophoresis showed that tubulin was the major molecular species present, although a number of minor components, possibly representing microtubule-associated proteins (MAPs), were present . The taxol procedure was also useful in obtaining other microtubule-containing structures such as the midbody or the cytoplasmic microtubule complex in interphase cells . The taxol procedure was also used to isolate mitotic spindles from HeLa cells . The HeLa spindles stained positively with an antibody specific for the 210 X 10(3) Mr microtubule-associated protein, indicating that the MAP was retained by the taxol procedure . The taxol procedure appears to be of great advantage in large-scale preparations of spindles for biochemical analysis.

Boll Soc Ital Biol Sper, 1984 Feb 28, 60(2), 351 - 4
{Comparison of various media and various methods for the growth and isolation of Pityrosporum pachydermatis}; Lorenzini R et al.; The pathogenic role of Pityrosporum pachydermatis in otitis externa of dogs and the related diagnostic problems are emphasized . We report results related to isolation, cultivation and identification of yeast . Agar nutritive glucosate with 1,5% of yeast extract has been showed as the best medium permitting identification in 24 hrs, associated with morfological test . Tween 80 integration (1%) to the medium permits to isolate lipolitic yeasts also.

J Biol Chem, 1984 Feb 25, 259(4), 2375 - 82
Regulation of asparagine-linked oligosaccharide processing . Oligosaccharide processing in Aedes albopictus mosquito cells; Hsieh P et al.; We have examined the synthesis and processing of asparagine-linked oligosaccharides from Aedes albopictus C6/36 mosquito cells . These cells synthesized a glucose-containing lipid-linked oligosaccharide with properties identical to that of Glc3Man9GlcNAc2-PP-dolichol . Results of brief pulse label experiments with {3H}mannose were consistent with the transfer of Glc3Man9GlcNAc2 to protein followed by the rapid removal of glucose residues . Pulse-chase experiments established that further processing of oligosaccharides in C6/36 cells resulted in the removal of up to six alpha-linked mannose residues yielding Man3GlcNAc2 whose structure is identical to that of the trimannosyl "core" of N-linked oligosaccharides of vertebrate cells and yeast . Complex-type oligosaccharides were not observed in C6/36 cells . When Sindbis virus was grown in mosquito cells, Man3GlcNAc2 glycans were preferentially located at the two glycosylation sites which were previously shown to have complex glycans in virus grown in vertebrate cells . These Man3GlcNAc2 structures are the most extensively processed oligosaccharides in A . albopictus, and as such, are analogous to the complex glycans of vertebrate cells . We suggest that determinants of oligosaccharide processing which reside in the polypeptide are universally recognized despite evolutionary divergence of the oligosaccharide-processing pathway between insects and vertebrates.

Biochem Biophys Res Commun, 1984 Feb 14, 118(3), 814 - 20
Biosynthesis of riboflavin . Enzymatic formation of the xylene moiety from {14C}ribulose 5-phosphate; Nielsen P et al.; We have studied the enzymatic formation of the xylene ring of riboflavin using cell extracts from the flavinogenic yeast Candida guilliermondii . 5-Amino-6-ribitylamino-2,4(1H,3H)-pyrimidinedione or its 5'-phosphate could serve as substrates . In addition, a pentose phosphate or pentulose phosphate was required . Experiments with {14C}ribulose 5-phosphate gave evidence for the incorporation of the ribulose carbon atoms except C-4 into the xylene ring of the vitamin.

J Biol Chem, 1984 Feb 10, 259(3), 1644 - 8
Purification and characterization of three elongation factors, EF-1 alpha, EF-1 beta gamma, and EF-2, from wheat germ; Lauer SJ et al.; Three elongation factors, EF-1 alpha, EF-1 beta gamma and EF-2, have been isolated from wheat germ . EF-1 alpha and EF-2 are single polypeptides with molecular weights of approximately 52,000 and 102,000, respectively . The most highly purified preparations of EF-1 beta gamma contain four polypeptides with molecular weights of approximately 48,000, 46,000 and 36,000, 34,000 . EF-1 alpha supports poly(U)-directed binding of Phe-tRNA to wheat germ ribosomes and catalyzes the hydrolysis of GTP in the presence of ribosomes, poly(U), and Phe-tRNA . EF-2 catalyzes the hydrolysis of GTP in the presence of ribosomes alone and is ADP-ribosylated by diphtheria toxin to the extent of 0.95 mol of ADP-ribose/mol of EF-2 . EF-1 beta gamma decreases the amount of EF-1 alpha required for polyphenylalanine synthesis about 20-fold . EF-1 beta gamma enhances the ability to EF-1 alpha to support the binding of Phe-tRNA to the ribosomes and enhances the GTPase activity of EF-1 alpha . Wheat germ EF-1 alpha, EF-1 beta gamma, and EF-2 support polyphenylalanine synthesis on rabbit reticulocyte ribosomes as well as on yeast ribosomes.

Nucleic Acids Res, 1984 Feb 10, 12(3), 1737 - 47
Nucleotide sequence of the 18S-26S rRNA intergene region of the sea urchin; Hindenach BR et al.; The DNA sequence which spans the internal transcribed spacers of a cloned ribosomal transcription unit from the sea urchin, Lytechinus variegatus, has been determined . The region extends from the conserved Eco RI site near the 3' end of the 18S rDNA to a Bam HI site in the 26S rDNA and includes 232 nucleotides coding for 18S rRNA, 367 nucleotides of internal transcribed spacer, 159 nucleotides coding for 5.8S rRNA, 338 nucleotides of internal transcribed spacer, and 505 nucleotides coding for 26S rRNA . The rRNA coding regions were identified by direct analysis of 3'-labeled 18S and 5.8S rRNA and 5'-labeled 5.8S rRNA, and by sequence homology of the 26S rDNA with yeast and vertebrate 26/28S rRNAs . The internal transcribed spacers are GC-rich, similar to those of vertebrates . The 5.8S and 5' 26S rDNA sequences support a proposed model for a structural domain of the yeast large subunit ribosomal RNA (Veldman et al . {1981} Nucleic Acids Res . 9, 6935-6952).

J Anim Sci, 1984 Feb, 58(2), 346 - 50
Effect of selenium on performance, serum selenium concentration and glutathione peroxidase activity in pigs; Adkins RS et al.; Pigs from sows fed a diet deficient in Se and low in vitamin E were fed a Torula yeast diet supplemented with 100 IU dl-alpha-tocopheryl acetate/kg of diet . Dietary treatments were levels of supplemental Se of 0, .025, .050, .075 or .100 ppm . Some death loss occurred in pigs receiving no supplemental Se at approximately 5 wk of age . Autopsy revealed liver and heart lesions typical of vitamin E-Se deficiency . Selenium supplement had no significant effect on average daily gain, feed intake or gain to feed ratio for the 4-wk experiment . Selenium status of pigs was determined by serum Se concentration and serum glutathione peroxidase (GSH-Px) activity . Serum Se increased linearly (P less than .01) with increasing supplemental Se . Serum GSH-Px activity increased linearly (P less than .01) and quadratically (P less than .05) with increasing supplemental Se . With time, the level of serum Se and GSH-Px activity decreased in unsupplemental pigs, but increased in pigs fed diets supplemented with Se and resulted in significant interactions (P less than .01) between dietary Se level and time on experiment . The correlation between serum Se concentration and GSH-Px activity was .81 (P less than .01).

Mol Biochem Parasitol, 1984 Feb, 10(2), 161 - 70
Sterols of Leishmania species . Implications for biosynthesis; Goad LJ et al.; The major sterol of promastigotes of stocks of Leishmania tropica, L . donovani and 3 subspecies of L . mexicana has been identified as ergosta-5,7,24(28)-trien-3 beta-ol; and of an L . major stock as ergosta-7,24(28)-dien-3 beta-ol . 24-Methylcholesta-5,7,22-trien-3 beta-ol and 24-ethylcholesta-5,7,22-trien-3 beta-ol were minor constituents, and traces of ergosta-5,7,22,24(28)-tetraen-3 beta-ol and a C27-diene were also recognized in some species . Lanosterol and 4,4-dimethylcholesta-8,24-dien-3 beta-ol were detected in all species studied, and squalene was identified in a stock of L . tropica . The sterol composition of members of the genus Leishmania and the sterol biosynthetic pathways it implies are characteristic of yeast and other fungi.

Clin Exp Immunol, 1984 Feb, 55(2), 303 - 12
Evidence for intrinsic cellular defects of 'complement' receptor-mediated phagocytosis in patients with systemic lupus erythematosus (SLE); Hurst NP et al.; Fc and 'complement'-mediated phagocytosis of pre-opsonized yeast has been studied in monocytes from 18 patients with SLE . Monocytes from nine of 18 patients had depressed complement-mediated phagocytosis (P = 0.0001, Fishers exact test) but only three of 18 had depressed Fc-mediated phagocytosis (NS, Fishers exact test) . Although reduced complement-mediated phagocytosis was correlated with increased frequency of disease manifestations (P less than 0.003, rank correlation) there was no correlation with serum DNA or C1q binding activity, C3 or C4 levels . Serum from SLE patients with depressed phagocytosis did not inhibit phagocytosis by normal monocytes . The data suggests the presence of intrinsic abnormalities of monocyte receptor function in SLE and the nature of the 'complement' receptors involved is discussed.

J Anim Sci, 1984 Feb, 58(2), 351 - 5
Effect of supplemental selenium on pancreatic function and nutrient digestibility in the pig; Adkins RS et al.; The effect of Se deficiency on pancreatic digestive enzyme activity and nutrient digestibility was studied by using pigs weaned at 3 wk of age from sows fed a diet deficient in Se and low in vitamin E . Pigs were fed a Torula yeast diet supplemented with 100 IU dl-alpha-tocopheryl acetate/kg of diet . Treatments were levels of supplemental Se of 0, .025, .050, .075 or .100 ppm . Apparent digestibility was determined at the end of the second and fourth week . Digestive enzyme activity, glutathione peroxidase (GSH-Px) activity and Se concentration were determined in pancreatic tissue at the end of the 4-wk experiment . Selenium concentration in the pancreas increased linearly (P less than .01) and quadratically (P less than .05) in response to increasing level of Se supplementation . The correlation between pancreas Se level and GSH-Px activity was .36 (P less than .05) . Supplementation of Se had no effect on pancreas weight, protein content or the activity of pancreatic trypsin, chymotrypsin, alpha-amylase or lipase . Apparent digestibility increased linearly for dry matter (P less than .01) and N (P less than .05) as Se supplementation increased . There was however, no significant effect on ether extract digestibility . Apparent digestibilities were higher (P less than .01) at 4 wk than those measured at 2 wk.

Proc Natl Acad Sci U S A, 1984 Feb, 81(4), 1084 - 8
Two initiation sites detected in the small s1 species of reovirus mRNA by dipeptide synthesis in vitro; Cenatiempo Y et al.; Reovirus mRNAs directed the synthesis of fMet dipeptides in a translation initiation system reconstituted from rabbit reticulocyte initiation and elongation factors, Artemia salina 80S ribosomes, yeast fMet-tRNAiMet and Escherichia coli3H-labeled aminoacyl tRNAs . As predicted from the GC(U,G) codon that follows the 5'-proximal AUG in half of the viral mRNA species, fMet-Ala was the predominant dipeptide product obtained in response to a mixture of mRNAs or to the separated size classes of medium (m) and small (s) mRNA . The four individual small mRNA species each directed the synthesis of an fMet dipeptide that was consistent with the utilization of the 5'-proximal AUG for initiation . In addition to fMet-Asp, the s1 mRNA also directed fMet-Glu synthesis indicative of initiation in a second reading frame at the 5'-penultimate AUG . The tripeptide fMet-Glu-Tyr was also synthesized from s1 mRNA, which further verified this second initiation site . mRNAs containing 5'-terminal GpppG were 10-15% as active as the corresponding m7G-capped templates . The dipeptide assay provides a rapid method for determining initiation sites in individual mRNAs or in mixtures of mRNAs.

South Med J, 1984 Feb, 77(2), 161 - 3
Significance of serum antibodies to Histoplasma capsulatum in endemic areas; George RB et al.; To evaluate the specificity of serologic tests for histoplasmosis in an endemic area, we studied sera from 104 consecutive healthy blood donors, testing for antibodies to either histoplasmin or Histoplasma yeast antigen, using complement fixation (CF), radioimmunoassay (RIA), and radial immunodiffusion (ID) . Twenty-five subjects (24%) had CF antibody titers of 1:8 or 1:16 to one or both antigens; none had titers above 1:16 . Nine subjects (9%) had RIA antibody titers of 1:8 or higher . No titers above 1:8 were found using the yeast antigen; five subjects had titers of greater than 1:16 to histoplasmin using RIA . No precipitin bands were found in any of these subjects . The results of four previous studies in endemic populations yielded similar or lower incidences of positive tests . CF antibody titers to either antigen, RIA antibody titers to yeast antigen of 1:32 or greater, or precipitin bands are rare in persons who live where histoplasmosis is endemic.

Med Hypotheses, 1984 Feb, 13(2), 139 - 51
Rationales for micronutrient supplementation in diabetes; McCarty MF et al.; Available evidence--some well-documented, some only preliminary--suggests that properly-designed nutritional insurance supplementation may have particular value in diabetes . Comprehensive micronutrient supplementation providing ample doses of antioxidants, yeast-chromium, magnesium, zinc, pyridoxine, gamma-linolenic acid, and carnitine, may aid glucose tolerance, stimulate immune defenses, and promote wound healing, while reducing the risk and severity of some of the secondary complications of diabetes.

Cell Biol Int Rep, 1984 Feb, 8(2), 147 - 50
Organization of microtubules in stabilized meristematic plant cells revealed by a rat monoclonal antibody reacting only with the tyrosinated form of alpha-tubulin; Wehland J et al.; A rat monoclonal antibody against yeast tubulin (clone YL 1/2; Kilmartin et al., 1982) that reacts specifically with mammalian alpha-tubulin carrying a carboxyterminal tyrosine residue (Wehland et al., 1983) was used to localize microtubules in plant cells derived from onion root apices (Allium cepa L.) . YL 1/2 reacted with all types of microtubular arrays known to occur in higher plant meristematic cells such as interphase cortical microtubules, pre-prophase bands, the mitotic spindle and phragmoplast microtubules . The specific labeling of microtubules in isolated cells from onion root tips by YL 1/2 indicates that plant cells like animal cells contain tubulin tyrosine ligase, the enzyme which posttranslationally modifies alpha-tubulin . This enzyme could be involved in the dynamic regulation of microtubular arrays in all eukaryotic cells.

Nucleic Acids Res, 1984 Jan 25, 12(2), 1069 - 75
The spatial organization of sequences involved in initiation and termination of eukaryotic DNA replication; Cook PR et al.; Nuclear DNA is looped by attachment to a matrix or cage . As this cage is the site of DNA synthesis, sequences in the loops must attach before they are replicated . We have tested whether sequences which initiate replication are usually out in the loop and attach only during S phase or whether they are attached but quiescent during most of the cell-cycle . Sequences which permit plasmids to replicate autonomously in yeast cells (ARS's) are strong candidates for initiating sequences . Four different human ARS's all map remote from attachment points to the HeLa nuclear cage . In addition a potential terminus of replication is also remote from the cage . We conclude that sequences involved in initiation are usually out in the loop and that DNA synthesis is initiated by their attachment.

Biochem J, 1984 Jan 15, 217(2), 561 - 71
Cytochrome c mediates electron transfer between ubiquinol-cytochrome c reductase and cytochrome c oxidase by free diffusion along the surface of the membrane; Froud RJ et al.; Ubiquinol oxidase can be reconstituted from ubiquinol-cytochrome c reductase (Complex III) and cytochrome c oxidase (Complex IV) whose endogenous phosphatidylcholine and phosphatidylethanolamine have been replaced by dimyristoylglycerophosphocholine . Phase transition of the lipid has no effect on Complex III and Complex IV activities assayed separately, but ubiquinol oxidase activity rapidly decreases as the temperature is lowered through the phase transition . A spin-labelled yeast cytochrome c derivative has been synthesized . Binding of the cytochrome c to liposomes demonstrates that only cardiolipin is involved under the conditions used for the ubiquinol oxidase experiments . In liposomes consisting of cardiolipin and dimyristoylglycerophosphocholine, e.s.r . (electron-spin-resonance) measurements show that rotational diffusion of cytochrome c is slowed in the gel phase of the latter lipid . We propose that the cytochrome c pool is bound to cardiolipin molecules, whose lateral and rotational diffusion in the bilayer is adequate to account for electron-transport rates.

Eur J Biochem, 1984 Jan 2, 138(1), 153 - 9
Evidence for an incomplete dolichyl-phosphate pathway of lipoglycan formation in Volvox carteri f . nagariensis; Muller T et al.; Crude membrane fractions from Volvox carteri in the presence of detergent and metal complexing agent catalyze the transfer of glucose from dolichyl phosphate glucose to branched dolichyl diphosphate chitobiosyl pentamannoside Dol-PP-(GlcNAc)2-(Man)5, a known intermediate of the lipid-mediated pathway of N-glycosylation of proteins, resulting in the formation of Dol-PP-(GlcNAc)2-(Man)5-(Glc)1 . Under the various conditions tested, neither Dol-P-Man nor other known mannosyl donors of the nucleoside-activated or lipid-activated type can serve as donor molecules for the elongation of the lipid-linked heptasaccharide . On the other hand, calf liver microsomes in similar experiments mannosylated the heptasaccharide further with Dol-P-Man up to a nonamannoside, Dol-PP-(GlcNAc)2-(Man)9 . A direct glucosylation of the acceptor, however, with Dol-P-Glc failed in this system . The (GlcNAc)2-(Man)5-(Glc)1, obtained after mild acid hydrolysis of the above glycolipid is not significantly split by an unspecific alpha-glucosidase from yeast . However, Volvox microsomes liberated most of the glucose indicating a specific glucosidase in the membranes of the alga . This enzyme does not act on (GlcNAc)2-(Man)9-(Glc)1, the usual protein-linked carbohydrate intermediate of trimming processes of N-glycosidic glycoproteins . The data on glycolipid formation let us postulate that in Volvox the normal N-glycosylation pathway differs from that found in higher plants and animals either by a lack of evolution or by mutation in the genes coding for the mannosyl transferases involved.

Eur J Biochem, 1984 Jan 2, 138(1), 67 - 75
Tertiary structure of animal tRNATrp in solution and interaction of tRNATrp with tryptophanyl-tRNA synthetase; Garret M et al.; Alkylation in beef tRNATrp of phosphodiester bonds by ethylnitrosourea and of N-7 in guanosines and N-3 in cytidines by dimethyl sulfate and carbethoxylation of N-7 in adenosines by diethyl pyrocarbonate were investigated under various conditions . This enabled us to probe the accessibility of tRNA functional groups and to investigate the structure of tRNATrp in solution as well as its interactions with tryptophanyl-tRNA synthetase . The phosphate reactivity towards ethylnitrosourea of unfolded tRNA was compared to that of native tRNA . The pattern of phosphate alkylation of tRNATrp is very similar to that found with other tRNAs studied before using the same approach with protected phosphates mainly located in the D and T psi arms . Base modification experiments showed a striking similarity in the reactivity of conserved bases known to be involved in secondary and tertiary interactions . Differences are found with yeast tRNAPhe since beef tRNATrp showed a more stable D stem and a less stable T psi stem . When alkylation by ethylnitrosourea was studied with the tRNATrp X tryptophanyl-tRNA synthetase complex we found that phosphates located at the 5' side of the anticodon stem and in the anticodon loop were strongly protected against the reagent . The alkylation at the N-3 position of the two cytidines in the CCA anticodon was clearly diminished in the synthetase X tRNA complex as compared with the modification in free tRNATrp; in contrast the two cytidines of the terminal CCA in the acceptor stem are not protected by the synthetase . The involvement of the anticodon region of tRNATrp in the recognition process with tryptophanyl-tRNA synthetase was confirmed in nuclease S1 mapping experiments.

Mol Cell Biochem, 1984, 61(2), 99 - 109
Origins of replication and gene regulation; Taylor JH; Eukaryotic chromosomes appear to consist of many replicons, the time of replication of which is probably controlled by specific origins . However, plasmids without specific eukaryotic origins may also replicate in some cells when injected into nuclei or transferred during transformation . The efficiency and the mechanisms of their initiation are still uncertain . A number of reports are cited which indicate that natural eukaryotic DNAs initiate their replication from specific origins . The nature of these origins are known in only a few instances and no general conclusions can yet be given about the nucleotide sequences involved . Short dispersed repeats of the Alu type appear to function as origins since they enhance the efficiency of replication of vector plasmids in Xenopus eggs . Certain sequences from a variety of eukaryotic DNAs also enhance the replicative potential of plasmids in yeast cells . The common features of such initiators or enhancers is uncertain . If dispersed repeats are origins in mammalian chromosomes, the number appears to be excessive . Either only a subset are functional, or the functional ones are only suborigins in larger replicons in which master origins (not yet isolated) function in the regulation of the timing of replication . Evidence is cited which indicates that the regulation of the time of replication of a gene or gene cluster is part of a regulatory system that makes the DNA available for transcription or leaves it in an inactive state . About one-half the DNA in mammalian cells is replicated in the first half of S phase (SE) . After a brief pause in mid-S phase, the remainder of the DNA is replicated in what is designated late S (SL) . The fractions replicated in SE and SL may vary in other phylogenetic groups, but wherever division of differentiated cells occurs such fractions are likely to be found . The following hypothesis is proposed . The DNA replicated in SL is suppressed in transcription, if it has the appropriate promoter regions, because the newly replicated DNA is complexed with proteins that suppress transcription . These proteins are only available during SL . Those genes replicated in SE are complexed with a different set of proteins which leave the promoter regions open for transcription when the appropriate regulatory molecules are available . In this way an inactive state or potentially active state can be transmitted from one cell generation to the next . Evidence is cited which indicates that genes which are active in all cells at some stage in the cell cycle are replicated in SE.(ABSTRACT TRUNCATED AT 400 WORDS)

Vet Immunol Immunopathol, 1984 Jan, 5(3), 289 - 95
Variations in sheep serum conglutinating and haemolytic activity for sheep erythrocytes sensitized by rabbit antibody; Jonas W et al.; Based on conglutinating and haemolytic reactions with sheep erythrocytes (E) sensitized by rabbit antibody (A), three types of sheep sera were encountered . Type 1 sera do not conglutinate or haemolyse sheep E-rabbit A . Type 2 sera failed to conglutinate or haemolytically active . Type 3 sera have both activities . Serum from one type 1 sheep still failed to conglutinate 5 days after venepuncture but was not haemolytically active (i.e., type 2) . Some sheep that initially had type 2 sera had, five days after an intraperitoneal injection of yeast cells, sera with conglutinating activity (type 3 sera) . Type 1, 2 and 3 sera all had haemolytic activity with human E-sheep A indicator cells . Pooled type 3 sera have the highest conglutinating titres with sheep E-rabbit A after 10 min incubation at 39 degrees C . At this stage, the haemolytic titres were very low . From 10 min, the conglutinating titres decreased whereas the haemolytic titres gradually increased until 80 min . Optimal conglutinating activity required less rabbit A to sensitize sheep E than did haemolytic activity.

Cell, 1984 Jan, 36(1), 145 - 54
Purified lupus antigen La recognizes an oligouridylate stretch common to the 3' termini of RNA polymerase III transcripts; Stefano JE; The small ribonucleoproteins recognized by anti-La sera consist mainly of RNA polymerase III products complexed with an antigenic cellular protein of 50 kd . A biochemical procedure for purifying the La protein from HeLa cells is described . The interaction of the isolated protein with a collection of model tRNA precursors, generated by ligation of specific oligonucleotides to the 3' terminus of yeast tRNAPhe, was studied . The most stable complexes are formed with adducts possessing three or four terminal uridylate residues . Addition of a terminal phosphate, fragmentation of the RNA, or substitution of other nucleotides reduce the affinity for the La protein . The preferred terminal sequence recognized and bound by La protein is homologous to the transcriptional termination signal for RNA polymerase III.

Nucleic Acids Symp Ser, 1984, (15), 125 - 8
Poly(U)-dependent polyphenylalanine and polytyrosine synthesis in vitro by a tRNATyr variant with an enzymatically altered anticodon, G-A-A; Nishikawa K et al.; A variant of T . utilis tRNATyr containing a base substitution (psi----A) in the middle position of the anticodon has been constructed by enzymatic procedures in vitro . This variant is unique in that it can accept both tyrosine and phenylalanine . This tRNA was shown to be active in transferring both tyrosine and phenylalanine into polypeptides in a cell-free, poly (U)-directed translation system from yeast . This result gives further support to the adapter hypothesis since tyrosine, attached to the variant tRNATyr with an anticodon G-A-A, is incorporated into polypeptides in response to poly (U) message.

Arzneimittelforschung, 1984, 34(9), 992 - 1001
{Animal experimental studies on the question of the interaction of paracetamol and acetylsalicylic acid}; Engelhardt G; The mutual effects of acetylsalicylic acid and paracetamol used in combination were studied in rats and mice . The antiexudative effects of acetylsalicylic acid in the edema tests on the hind paw of the rat, the antipyretic activity against yeast-fever of the rat and the analgesic effect on the inflamed hind paw of the rat was potentiated by paracetamol given simultaneously . The biosynthesis of prostaglandins in vitro by an enzyme preparation from bovine seminal vesicles was inhibited by acetylsalicylic acid and paracetamol in additive synergistic mode . The hepatotoxicity of paracetamol in mice measured by an increase of GOT in serum is antagonized by acetylsalicylic acid in dose-dependent manner . The suppression of the hepatotoxicity of paracetamol by acetylsalicylic acid in mice is not the consequence of interaction during absorption . The 3H-content of the liver following an oral dose of 3H-paracetamol is not influenced by acetylsalicylic acid given in combination with paracetamol . The mechanism of interactions between acetylsalicylic acid and paracetamol is discussed.

Sabouraudia, 1984, 22(2), 109 - 16
Paecilomyces lilacinus (Thom) Samson, from systemic infection in an armadillo (Dasypus novemcinctus); Gordon MA; Paecilomyces lilacinus was recovered in culture from pulmonary lesions and other internal organs of a captive armadillo . Impression films and smears of the lung tissue revealed numerous budding yeast-like cells, many of them appearing encapsulated and resembling intracellular forms of Histoplasma capsulatum . Yeast-like forms developed in culture on appropriate media at 35 degrees C . The armadillo isolate was compared in culture at both 27 degrees C and 35 degrees C with bamboo rat and human isolates of Penicillium marneffei . The two species were also subjected to serologic and pathogenicity studies . Sensitivities of the armadillo isolate to five antifungal agents in vitro were consistent with those of three human isolates of this species but contrasted sharply with those of P . marneffei.

Folia Microbiol (Praha), 1984, 29(1), 8 - 13
A sulfate, sulfite and thiosulfate incorporating system in Candida utilis; Alonso A et al.; Sulfate, sulfite and thiosulfate incorporation in the yeast Candida utilis is inhibited by extracellular sulfate, sulfite and thiosulfate and by sulfate analogues selenate, chromate and molybdate . The three processes are blocked if sulfate, sulfite, thiosulfate, cysteine and homocysteine are allowed to accumulate endogenously . Incorporation of the three inorganic sulfur oxy anions is inactivated by heat at the same rate . Mutants previously shown to be defective in sulfate incorporation are also affected in sulfite and thiosulfate uptake . Revertants of these mutants selected by plating in ethionine-supplemented minimal medium recovered the capacity to incorporate sulfate, sulfite and thiosulfate . These results taken together with previous evidence demonstrate the existence of a common sulfate, sulfite and thiosulfate incorporating system in this yeast.

Trans Am Clin Climatol Assoc, 1984, 96, 67 - 78
The effects of essential fatty acid deficiency on pulmonary alveolar macrophage function; Balint JA et al.; Male rats were maintained for periods of up to 16 weeks on a fat free diet which was supplemented with either 4% tripalmitin (essential fatty acid {EFA} deficient) or with 4% safflower oil (SAFF, control) . Pulmonary alveolar macrophages (PAM) were obtained by lung lavage . PAM from EFA deficient rats had reduced phagocytic activity and capacity . Intracellular killing of ingested yeast was also reduced by EFA deficiency . The activity of acid phosphatase, beta-glucuronidase and cathepsin D from PAM was not altered by dietary treatment . Transmission electron microscopy failed to show any consistent morphologic differences between PAM from EFA deficient and SAFF animals, but did confirm the decreased phagocytosis by PAM from EFA deficient rats . However, scanning electron microscopy did show loss of pseudopodia in PAM from EFA deficient rats . EFA deficiency was demonstrated by analyzing the methyl esters of the fatty aids from the total lipid extract of PAM . The arachidonate content was decreased while the eicosatrienoate content was increased in PAM derived from rats fed the EFA deficient diet . In an effort to elucidate further the mechanism of action of EFA deficiency in impairing phagocytosis by PAM, inhibitors of various reactions which lead to oxygenated derivatives of arachidonate were studied using PAM from chow fed rats . Some of these inhibitors were effective in diminishing phagocytosis . Furthermore, PAM from these preparations when fixed in suspension and examined with scanning electron microscopy showed morphological changes similar to those seen in EFA deficiency . This similarity of surface ultrastructural changes suggests that EFA deficiency may impair phagocytic function of PAM by reducing availability of an oxygenated derivative of arachidonic acid.

Antonie Van Leeuwenhoek, 1984, 50(5-6), 807 - 13
Morphological taxonomy of little-differentiated Hyphomycetes; de Hoog GS et al.; Morphological taxonomy of simple Hyphomycetes is complicated by the frequent occurrence of pleoanamorphism . In some groups of yeast-like fungi, uncommon synanamorphs are diagnostic . Differences in conidiogenesis do not always delimit natural groups . Some nomenclatural problems are mentioned, with an emphasis on the need of neotypification . Prospects are sketched for future taxonomic research.

Gen Pharmacol, 1984, 15(6), 535 - 9
Use of the artificial beta cell (ABC) in the assessment of peripheral insulin sensitivity: effect of chromium supplementation in diabetic patients; Elias AN et al.; The artificial beta cell (ABC), a closed-loop insulin delivery system, was used to determine insulin sensitivity . Progressively increasing glucose loads were administered after initial stabilization of the blood glucose at euglycemic levels such that the serum C-peptide was suppressed . The amount of insulin required to maintain euglycemia was considered a measure of sensitivity to insulin . Six stable maturity onset diabetics were studied before and after supplementation with chromium-rich brewer's yeast . All patients demonstrated an increase in sensitivity to insulin as indicated by a decrease in the fasting blood glucose concentration and a decrease in insulin requirement during the glucose challenge (P less than 0.02) . The data obtained support the hypothesis that chromium or some other factor(s) present in brewer's yeast potentiates the peripheral effects of insulin . It remains to be established whether this effect occurs at the receptor or post-receptor level.

Zh Nevropatol Psikhiatr Im S S Korsakova, 1984, 84(9), 1330 - 3
{Dimexide (dimethylsulfoxide) in the treatment of multiple sclerosis}; Zingerman LIa; Thirty-four patients with multiple sclerosis were treated with dimexide . The use of the drug was found to be desirable, since it had a positive effect on immunity and antiallergic and reparative action on the injured tissues . The treatment proved most effective in patients with a remitting course of the disease . In patients with a rapidly progressive course, the improvement was unstable . No side-effects were observed . The beneficial therapeutic effect of dimexide may be explained by remyelinization, a reduction in the edema and neurodynamic improvement.

Dev Comp Immunol, 1984 Summer, 8(3), 579 - 88
Endocytosis of a mannose-terminated glycoprotein and formaldehyde-treated human serum albumin in liver and kidney cells from fish (Salmo alpinus L.); Smedsrud T et al.; The uptake and degradation of a mannose-terminated glycoprotein, yeast invertase, in char (Salmo alpinus L.) tissue was studied after intravenously injection of the 125I-labelled protein . 125I-labelled formaldehyde-treated human serum albumin (fHSA) and native HSA was also injected for comparison . Labelled invertase was rapidly cleared from blood and at about the same rate as labelled fHSA (at 8 degrees C) . Approximately 50% of the initial concentration remained in blood 15 min after the injection of the ligands . Acid soluble degradation products appeared in the circulation about 60 min after the injection of the proteins . 125I-labelled invertase was recovered in the liver, pronephros and kidney . The clearance of labelled invertase from blood and the uptake in the organs were inhibited by co-injection of excess unlabelled invertase . fHSA was taken up in the pronephros and kidney tissue, while HSA was not taken up in any organs . In vitro degradation of the labelled ligands was studied in isolated pronephros cells, which had taken up the proteins in vivo . The degradation of invertase in isolated cells was partly inhibited by ammonium chloride . Ammonium chloride and chloroquine inhibited degradation of fHSA, but not leupeptin . These results together suggest that invertase and fHSA were taken up in the organs described by the receptor-mediated endocytosis . The degradation was partly or wholly lysosomal.

Pharmatherapeutica, 1984, 3(10), 682 - 5
Oxiconazole in the treatment of vaginal candidiasis: single dose versus 3-day treatment with econazole; Gouveia DC et al.; Fifty-one women with vaginal candidiasis were included in an open randomized study to compare the efficacy and local tolerance of a single-dose treatment with 600 mg oxiconazole (one vaginal tablet) with that of a 3-day treatment with econazole (one 150 mg ovule daily) . Twenty-five patients were treated with oxiconazole and the remaining 26 received econazole . Patients were followed-up 1 week and 4 to 5 weeks after the initial examination and details were recorded of the results of yeast culture, the clinical signs and symptoms present and any signs of intolerance . A cure rate of 92% was achieved in both treatment groups and only 1 patient (econazole group) of the 4 treatment failures (2 in each group) did not respond to a subsequent second course with the same treatment schedule . Vaginal burning was the only reported local side-effect in 5 patients of the oxiconazole group and in 6 of the econazole group.

J Interferon Res, 1984 Summer, 4(3), 315 - 27
Markedly enhanced production of gamma interferon in murine T lymphocytes treated with lentil lectin and the diterpene ester, mezerein; Taylor JL et al.; Gamma interferon (IFN-gamma) was induced in murine splenocytes first stimulated to grow by concanavalin A (Con A) and subsequently treated for 3 h with the diterpene ester, mezerein (MZN) and then with lectin from Lens culinaris for 24 h . Yields as high as 60,000 u/ml were obtained in cells from either male or female, random-bred, white Swiss mice or inbred C67B1/6 mice . Antibody to Thy 1.2 surface antigen completely obliterated the mouse gamma interferon (MuIFN-gamma) response, whereas anti-Lyt 1.2 and anti-Lyt 2.2 each destroyed a portion of the lymphocyte population responsible for MuIFN-gamma production . Kinetic analysis of production and release showed that IFN was detectable in culture fluids within 4 h after treatment with very little IFN remaining cell-associated (less than 10%) . A simple, rapid, and economical two-step purification procedure involving ammonium sulfate fractionation and yeast RNA affinity chromatography resulted in as much as 770-fold purification to achieve specific activities greater than 10(7) u/mg protein . The purified MuIFN-gamma was shown to be predominantly acid-labile, inactivated by sodium dodecyl sulfate (SDS), and neutralized by antiserum to MuIFN-gamma . Approximately 10% of the MuIFN-gamma was acid-stable and SDS-resistant, but was still neutralized by anti-MuIFN-gamma serum . Two molecular weight peaks of about 40 and 20 kD were demonstrated by SDS-polyacrylamide gel electrophoresis . Isoelectric focusing in polyacrylamide slab gels gave a relatively heterogeneous band of activity between pH 5.5 and 6.5 . The mechanism by which the combination treatment described enhances MuIFN-gamma production so markedly remains unknown, but the degree of enhancement is greater than additive.

J Biol Response Mod, 1984, 3(2), 132 - 7
Induction of neutrophilic differentiation of human promyelocytic leukemic cells by branched-chain carboxylic acid anticonvulsant drugs; Fischkoff SA et al.; The anticonvulsant drug 1-methyl-1-cyclohexanecarboxylic acid ( MCCA ) has been shown to cause maturation of murine neuroblastoma cells in vitro at concentrations that are pharmacologically achievable . HL-60 human promyelocytic leukemia cells cultured with this drug underwent a dose-dependent decrease in growth . Similarly, neutrophilic differentiation, based on morphologic criteria and the acquisition of the ability to reduce nitroblue tetrazolium and phagocytose yeast, was observed . Valproic acid, a clinically available anticonvulsant that is chemically related to MCCA , likewise inhibited growth and promoted maturation of HL-60 cells, although only at concentrations above the recommended therapeutic blood levels . MCCA was additive in its ability to induce differentiation of HL-60 with retinoic acid, another compound that induces differentiation at pharmacologic concentrations . MCCA , or similar branched-chain fatty acids, may be useful in the treatment of human leukemia, particularly in combination with other differentiation-inducing drugs.

Scand J Urol Nephrol Suppl, 1984, 86, 237 - 40
Single dose of tinidazole in the treatment of vaginal discharge; Paavonen J et al.; Seventy-nine unselected women whose main symptom was abnormal vaginal discharge were seen at the Department of Obstetrics and Gynecology at Helsinki University Central Hospital between October 1980 and September 1981 . All patients and their current sex partners were randomly treated either with a 2-gram single dose of tinidazole or with identical placebos . Those excluded were 9 patients who did not attend the follow-up examination, 15 who had specific cervicovaginal infections (6 with chlamydial infection, 4 with trichomoniasis and 5 with yeast infection), and 22 with normal Doderlein flora seen on vaginal Pap smear . Response to the treatment was analysed among the remaining 33 women, of whom 17 received tinidazole and 16 placebo . The symptoms and signs that best discriminated between those who had normal Doderlein flora and those who had non-Doderlein flora were malodour, other than white colour of the discharge, and the presence of clue cells on Pap smear . When these findings were used to evaluate the response to treatment, tinidazole proved to be more effective than placebo . Disappearance of clue cells best (p less than 0.01) discriminated between tinidazole and placebo.

Sabouraudia, 1984, 22(6), 471 - 6
An investigation of the role of true hypha production in the pathogenesis of experimental oral candidosis; Martin MV et al.; From 427 cases of human oral candidosis 2135 yeast clones were screened for the presence of germ tube-negative C . albicans and variants that formed only pseudohyphae in serum; one strain of each was found . The pathogenic potential of the serum-pseudohyphal and germ tube-negative C . albicans variants was investigated in the oral cavity of the rat; both variants failed to induce palatal candidosis, in contrast to a germ tube-positive C . albicans control strain . The pathogenic potential of C . albicans strains appears to be dependent on the formation of true germ tubes.

J Biol Stand, 1984 Jan, 12(1), 67 - 74
The standardization of a 'spot-test' ELISA for the rapid screening of sera and hybridoma cell products I . The determination of the optimum buffering system; McCullough KC et al.; Using the different commercially available enzyme-linked immunosorbent assay (ELISA) plates, several sources of albumin were tested along with Tween 20 as supplements to the diluting buffer in ELISA for their ability to minimize non-specific reactions . There was an obligate requirement for Tween 20 (0.05%) but the different albumin sources had varied effectiveness on each of the different ELISA plates . In general, however, the optimum buffering system was concluded to be phosphate-buffered saline supplemented with 0.05% (v/v) Tween 20 and 3% (w/v) lactalbumin yeast hydrolysate or bovine serum (plasma) albumin fraction V.

Infect Immun, 1984 Jan, 43(1), 320 - 5
Correlation of elastase production by some strains of Aspergillus fumigatus with ability to cause pulmonary invasive aspergillosis in mice; Kothary MH et al.; Seventy-five strains of Aspergillus fumigatus were screened for production of elastase in liquid and agar media containing elastin in yeast carbon base buffered with 0.05 M borate, pH 7.6 . Of 71 strains which cleared elastin in agar plates, 33 produced elastase in liquid medium, as measured spectrophotometrically with elastin-Congo red . Six strains producing elastase and four nonproducers were tested for ability to cause invasive aspergillosis in immunocompromised mice (six mice per strain) . All 36 mice exposed to elastase-producing strains died within 48 to 96 h . Lung tissue from dead mice showed hyphae and necrosis of the alveoli . Lungs of mice exposed to spores of strains not producing elastase showed few germinated spores and no destruction of alveoli . These results indicate that elastase may be significant in the invasion process.

Zentralbl Bakteriol Mikrobiol Hyg {B}, 1984 Jan, 178(5-6), 527 - 34
{Detection of enteroviruses in seawater and beach sand}; Nestor I et al.; In the years from 1975 to 1978 investigations were carried out to detect enteroviruses in the sea-water and in the sand of the beaches of the Rumanian Black Sea coast, in zones with and without waste water discharge . The quantities of enteroviruses found in the seawater and in the sand of the beaches were lower than those verified in other countries . For the identification of viruses we used two concentration methods simultaneously (polyelectrolyte PE60 Monsanto USA and brewer's yeast cells) as well as two methods of isolation (in cell cultures and with newborn mice) . The incidence of enteroviruses depended on the season, with no viruses present in the sea-water and in the sand of the beaches during the periods considered to be outside the holidaying season . The discharge of purified sewage into the sea was not attended by considerable viral contamination of the sea-water and of the beaches.

J Exp Med, 1984 Jan 1, 159(1), 244 - 60
Local opsonization by secreted macrophage complement components . Role of receptors for complement in uptake of zymosan; Ezekowitz RA et al.; We have examined the role of macrophage (M phi plasma membrane receptors for the cleaved third complement component (iC3b; CR3) and mannosyl, fucosyl terminated glycoproteins (MFR) in uptake of unopsonized zymosan . Monoclonal antibodies against CR3, M1/70 (Mac-1) and MO1, each inhibited approximately 50% of uptake of 125I-zymosan by murine and human M phi, respectively . Yeast mannan inhibited 0-50% of zymosan uptake in various M phi, in parallel with their expression of MFR . We demonstrated that M phi were the source of C3 in our assay and that the activity of other components of the complement system, namely a C3 convertase, factor I, and a factor I cofactor were also present in serum-free cultures of human monocytes . Macrophage C3 was deposited rapidly, within 10 min, on the zymosan particles and mediated binding, ingestion, and stimulation of superoxide release in BCG-activated and thioglycollate-elicited peritoneal M phi via CR3 . Local secretion of complement proteins by M phi themselves can therefore opsonize pathogens and cells able to activate the alternative pathway and effect their destruction.

Ann Dermatol Venereol, 1984, 111(12), 1081 - 5
{Pityrosporum ovale in keratotic lesions of seborrheic areas}; Bourlond A et al.; The Pityrosporum ovale (PO) is demonstrated in various percentages of the keratotic lesions of the seborrheic areas depending upon the techniques . The accumulation of horny material most probably enhances the multiplication of the yeast and makes its demonstration easier . The PAS technique prevails upon the mycological ones (77 p . 100 versus 50-65 p . 100) . PO proliferates only in the keratin layer; there are isolated spores, small clusters or even large stratified colonies . On the other side the prevalence of PO does not differ significantly whatever the ultimate diagnosis of seborrheic keratosis, actinic keratosis, naevocellular nevus or vial wart . Its multiplication within the keratin layer of the lesions does not make any difference as far as the lymphocytic infiltration of the dermis is concerned.

Acta Physiol Pharmacol Latinoam, 1984, 34(1), 25 - 30
Effects of different diets on voluntary consumption of ethanol in UChA and UChB rats; Mardones J et al.; Based on the decrease of voluntary consumption of ethanol observed in rats of UChA (low ethanol consumer) and UChB (high ethanol consumer) strains, coincident to the use of a new issue of a commercial diet, the effects of two new diets devoid of animal food were studied . One of these diets (Diet 3) induced an increase of the voluntary consumption of ethanol in a proportion of UChA rats, in such a way that its frequency distribution curve resulted in a bimodal one because of the presence of individuals which drank as much alcohol as UChB rats . This diet is a useful tool for the study of eventual nutrimental factors which decrease ethanol intake.

J Inorg Biochem, 1984 Jan, 20(1), 69 - 78
Effects of 2-formylpyridine monothiosemicarbazonato copper II on red cell components; Antholine WE et al.; 1-Formylpyridine monothiosemicarbazonato copper II (CuL+) is readily taken up by red cells and is initially bound to glutathione and hemoglobin . Glutathione was depleted within 5 hr of incubation, presumably by oxidation mediated by CuL+ and O2 with concomitant generation of toxic oxygen species . Cupric ion was slowly transferred from CuL+ to hemoglobin within about 7 hr and hemoglobin was oxidized until the major form prevailing after 10 hr was alpha 2 beta 2+ . Little increase in hemolysis due to addition of CuL+ dissolved in the radical scavenger dimethyl sulfoxide was observed with prolonged incubation . Strong inhibition of red cell hexokinase by CuL+ was observed when the enzymes in red cell lysates and hemoglobin-free red cell lysates were examined . CuL+ was also an effective inhibitor of yeast hexokinase . However, the inhibitory effect of CuL+ within the red cells was less pronounced . It is suggested that even though intracellular accumulation of CuL+ creates an oxidizing environment and is potentially capable of inhibiting thiol enzymes such as hexokinase, protective effects are exerted in the red cell by the presence of hemoglobin, of radical scavengers, and of high levels of enzymes that detoxify toxic oxygen species.

Int Arch Allergy Appl Immunol, 1984, 75(1), 32 - 7
Histamine release from mast cells during phagocytosis and interaction with activated neutrophils; Coble BI et al.; Histamine release from rat peritoneal mast cells was evaluated during interaction with IgG, C3b-bi and Concanavalin A-opsonized yeast particles and activated neutrophils . In contrast to others we could show no phagocytic uptake of yeast particles attaching to Fc, or Concanavalin A receptors on the mast cell . Attached yeast particles opsonized with C3b/bi were also poorly ingested (less than 10% of the attached particles.) Neither could we detect any significant histamine release . If, however, neutrophils were added to the mast cell-yeast particle complex, histamine release was induced particularly in the presence of Concanavalin A-yeast . We furthermore showed that phorbol myristate acetate-activated neutrophils and eosinophils initiated mast cell degranulation and histamine release . This release is not dependent on myeloperoxidase, but on other oxidative metabolites, since myeloperoxidase-deficient neutrophils also induce histamine release . These experiments show that mast cells exposed to immune complexes and activated neutrophils or eosinophils may augment the inflammatory response.

Exp Cell Res, 1984 Jan, 150(1), 36 - 46
pH in the endosome . Measurements during pinocytosis and receptor-mediated endocytosis; Geisow MJ et al.; By fluorescence spectroscopy, the average pH within endocytic compartments was determined during endocytosis of fluorescein conjugates by macrophages and hepatocytes . In mouse macrophages and hepatocytes fluorescein conjugates taken up either in the fluid phase or by binding to cell surface receptors were rapidly transferred to an acidic compartment (pH 5-5.5) . The half-time for this process was generally less than 4 min . The pH within yeast-containing phagosomes was also rapidly reduced to similar levels, following a unique and transient increase . In each case, the acid endosomal compartments involved probably do not contain lysosomal enzymes . When fluorescein conjugates of asialoglycoproteins were internalised by hepatocytes at 20 degrees C, no proteolysis occurred within the acidic endosome until the temperature was raised . Fluorescein conjugates of concanavalin A (conA) and polylysine were relatively more slowly internalised by macrophages . The half-times for uptake, estimated by fluorescence change, were comparable with the turnover time for bulk plasma membrane . The relatively high average pH experienced by these conjugates indicated that a small proportion of these non-specific cell-surface labels was always in contact with the extracellular medium.

Eur J Immunol, 1984 Jan, 14(1), 47 - 52
Marginal zone B cells express CR1 and CR2 receptors; Gray D et al.; Opsonized yeast is known to bind strongly to the marginal zones in frozen sections of rat spleen (Kumararatne, D . S . et al., Eur . J . Immunol . 1981 . 11: 858) . This study reports an analysis of the cells involved in this binding . Sheep red cells coated respectively with C3b, C3bi or C3d were used as indicator cells . These showed homogeneous binding of both C3b and C3bi to marginal zones and germinal centers . C3d-coated red cells bound in a uniform speckled pattern to marginal zones . They also bound to germinal centers and the small lymphocyte zones of the follicles . Selective depletion experiments were undertaken to show that binding to marginal zones was a property of the IgM+ and IgD- B cells characteristic of this area . Binding to germinal centers was attributable to follicular dendritic cells . The C3d receptors in follicles were shown to be on IgM+ and IgD+ small B lymphocytes.

Allerg Immunol (Leipz), 1984, 30(4), 225 - 9
Microbicidal activity and superoxide production by macrophages in aging rats; Ganguly R et al.; This study examined the effects of aging on the immunobiological properties of alveolar macrophages in young adult and aged rats . Macrophages, obtained by lung lavage, were enumerated and tested in vitro for yeast phagocytosis and fungicidal activities . Superoxide anion (O2-) generation stimulated by opsonized zymosan was compared between the two groups . Total numbers of alveolar cells and the differential cell distribution were comparable in both the groups . The difference between the groups was not statistically significant, even though the yeast phagocytic capacity in the cells from senescent animals was consistently higher than in the controls . This may be due to the wide degree of variability observed in the aged animals . However, cell fungicidal activity was statistically lower in the aged animals at 90 minutes of incubation which correlated with a significant decline in the O2- production . Aging may adversely affect some of the immunobiologic functions of lung macrophages and thus compromise host defense mechanisms in the respiratory tract.

Sabouraudia, 1984, 22(4), 331 - 9
Phialophora parasitica, an emerging pathogen; Weitzman I et al.; Monoconidial cultures derived from 12 clinical and environmental isolates of Phialophora parasitica were compared with respect to morphologic and physiologic characteristics and response to antifungal agents . No yeast cells were seen in 1- and 3-week-old slide culture preparations . Also, not all of the distinguishing characteristics of this species were displayed by all isolates on all media examined . Although the isolates grew on Sabouraud agar with chloramphenicol and cycloheximide, some inhibition was observed . All cultures were strongly urease-positive and hydrolyzed casein and starch; most decomposed tyrosine but not gelatin . All but one environmental isolate grew well at both 23 and 37 degrees C, but none grew at 40 degrees C . In the sensitivity testing the isolates did not vary much in their response to each drug, although some anomalies were observed . Amphotericin B and miconazole had minimum inhibitory concentrations in the low sensitivity range (2.0-8.0 and 2.5-10 micrograms m-1 respectively), for most isolates, and most isolates were resistant to both 5-fluorocytosine and ketoconazole . Limited observations were made on three other Phialophora species which might be confused with P . parasitica.

Int Rev Cytol, 1984, 91, 141 - 86
Transport of proteins into mitochondria; Doonan S et al.; There is still much that is obscure concerning the transport of proteins into or through the mitochondrial membrane systems . In addition, as pointed out previously, it is unlikely that the details of the process are the same for proteins destined for different compartments of the organelle . A brief summary of the process for matrix proteins might be as follows: The proteins are synthesized on free polysomes as precursors of higher molecular weight than the native forms . These precursors are liberated into the cell cytosol and subsequently translocated into the mitochondria . This timing might be different in yeast under some circumstances, synthesis being completed in association with the mitochondria . The precursors interact with a receptor in the outer mitochondrial membrane interaction being mediated by the presequences of the precursors . The presequences therefore act as addressing signals as well as possibly playing a role in one or all of (a) solubilization of precursors, (b) prevention of premature assembly into multimeric structures, or (c) maintenance of nonnative configurations required for transport . Interaction occurs with a second receptor, this time in the inner membrane of the mitochondria, interaction being with multiple sites in the polypeptide chain . Transport across the inner membrane then occurs, this transport depending on a transmembrane electrochemical gradient of which the proton component is the essential part . Transport is accompanied or followed by proteolysis of the prepiece, and formation of the native structure . While steps 1 and 2 of this sequence can be considered well established, the remaining steps are still poorly understood or purely hypothetical . Nevertheless, this sequence of events is consistent with known facts about the process and provides a framework for future investigations.

J Hyg Epidemiol Microbiol Immunol, 1984, 28(4), 471 - 8
Cell-mediated resistance induced by axenic amoebal RNA--protein fraction; Sharma GL et al.; Immunologically potent, RNA protein fraction could be isolated from axenic Entamoeba histolytica (NIH: 200) using a phenol extraction method . Immunization of animals with amoebal RNA protein (Eh RNA) fraction evoked an intense cellular sensitization . In immunized animals the percent leucocyte migration inhibition was found to be 73.7 +/- 6.9 . In control groups where the animals were treated with Freund's complete adjuvant, yeast RNA and amoebal RNA treated with RNase, the percent leucocyte migration inhibition was 15.8 +/- 2.9, 16.8 +/- 6.2 and 14.0 +/- 4.0 respectively . Anti Eh RNA antibodies (haemagglutinins and precipitins) could not be demonstrated in immunized animals at any stage of observations . The cellular immunity could be maintained at its high level by giving three immunization doses of Eh RNA and also a high degree (95.45%) of protection against experimental challenge with virulent strain of E . histolytica could be obtained in immunized animals . The immunogenic and protective capabilities of Eh RNA were found to be specific since they were abolished by the RNase treatment . Further, the yeast RNA failed to elicit immunity.

Allerg Immunol (Leipz), 1984, 30(4), 251 - 4
Influence of chemotherapy on the phagocytic activity of mononuclear cells in patients with Hodgkin's disease; Suss J et al.; The phagocytic activity of mononuclear cells (MNC) was measured in 18 healthy blood donors and 16 patients with Hodgkin's disease (HD) before and after a new chemotherapeutic cycle (Cyclophosphamide - Vincristine - Prednisone - Procarbazine, CVPP; Adriamycin - Bleomycin - Vinblastine - Decarbazine, ABVD) using radiolabeled, opsonized yeast cells . The phagocytic activity of the MNC of HD patients was found to be significantly higher than that of the healthy controls . In patients in whom chemotherapy induced remission or partial remission, phagocytic activity of MNC was also reduced significantly, while phagocytic activity was increased in two patients, whose clinical course was not influenced by therapy.

Nature, 1983 Dec 22-1984 Jan 4, 306(5945), 803 - 6
The protein products of the myc and myb oncogenes and adenovirus E1a are structurally related; Ralston R et al.; Structural and functional homologies have been found among proteins encoded by several retroviral oncogenes, demonstrating the existence of families of these genes . Because the retroviral oncogenes have cellular homologues, the existence of similar families among these 'cellular oncogenes' is also implied (for a review, see ref . 2) . Cellular genes belonging to these families have been found in such evolutionarily distant species as humans, fruit flies, nematodes and brewer's yeast (E . Scolnick and S . Reed, personal communications), consistent with the hypothesis that these genes have evolved from a small number of ancestral sequences . We extend these observations by showing here that the proteins encoded by the oncogenes myc, myb and adenovirus E1a are structurally related . Our findings suggest that oncogenes of RNA and DNA tumour viruses may in at least some instances share evolutionary origins and function according to common principles.

Mycopathologia, 1983 Dec 1, 84(1), 21 - 30
The effects of heat on Sporothrix schenckii in vitro and in vivo; Hiruma M et al.; In order to clarify the mechanism of action of topical thermotherapy on sporotrichosis, the effects of heat on Sporothrix schenckii in vitro and in vivo were investigated by observing the percentage germination and the ultrastructure . When the spores were heated to 42 degrees C, it took 10 hr with the conidia, 2 hr with the yeast-like cells and 1 hr with the spores in vivo to reduce the germination rates to 10% . The percentage germination curves were reduced slowly at first but later exponentially . Changes in the ultrastructure became evident in 2 hr with the yeast-like cells and in 8 hr with the conidia . The ribosome count declined and amorphous dense materials appeared in the cytoplasm and mitochondria . In vivo, the outstanding feature of the heated spores was the diversity of internal ultrastructural changes encountered and morphological changes . These were observed at 1 hr post treatment.

J Biochem (Tokyo), 1983 Dec, 94(6), 1827 - 32
Purification and characterization of an alpha-mannosidase of sea-squirt; Shigeta S et al.; An alpha-mannosidase was isolated from the extract of acetone powder of CHCl3-treated internal organs of the sea-squirt, Styela plicata . The enzyme was purified 4,110-fold in 11% yield . The preparation was fairly homogeneous on disc and SDS-polyacrylamide gel electrophoreses and Sephadex G-200 chromatography . The enzyme had an estimated molecular weight of 275,000 by gel chromatography and 70,000 by SDS-polyacrylamide gel electrophoresis, and was therefore considered to be a tetramer . The optimum pH for the enzyme activity was 3.4 but the stable pH range was from 4 to 6 . The isoelectric point was 5.0 . The enzyme was activated by Zn2+ but inhibited by Cu2+, Fe2+, Hg2+, and EDTA . The isolated enzyme released mannose not only from stem bromelin glycopeptide and ovalbumin glycopeptide but also from yeast mannan.

Biochem J, 1983 Dec 1, 215(3), 605 - 12
Purification and characterization of ribonucleases from human seminal plasma; Lee CL et al.; Four ribonucleases (RNAases I-IV) have been purified to homogeneity from human seminal plasma by precipitation with 40-75%-satd . (NH4)2SO4, followed by chromatographies on concanavalin A-Sepharose 4B, DEAE-cellulose phosphocellulose, agarose-5'-(4-aminophenylphospho)uridine 2'(3')-phosphate (RNAase affinity column) and Sephadex G-75 or G-100 . The homogeneity of these RNAases was confirmed by polyacrylamide-gel electrophoresis . Mr values for these purified RNAases were 78 000, 16 000, 13 300 and 5000 as estimated by gel filtration . Enzyme activities of RNAases I, III and IV were inhibited by Mn2+, Zn2+ and Cu2+ and activated by Na+, K+, Ba2+, Mg2+, Fe2+ and EDTA, whereas that of RNAase II was inhibited by Ba2+, Mg2+, Fe2+, Mn2+, Zn2+ and Cu2+ and activated by Na+, K+ and EDTA . RNAases I, II and IV demonstrated a higher affinity for poly(C) and poly(U) or yeast RNA, whereas RNAase III preferentially hydrolysed poly(U) over poly(C) and yeast RNA . In the presence of 5 mM-spermine, RNAase I was dissociated to a low-Mr (5000) enzyme with an increase in total RNAase enzymic activity . Xenoantiserum to each RNAase was raised and evaluated by immunoprecipitation and immunohistochemical methods . Anti-(seminal RNAase III) antiserum showed no immunological cross-reaction with RNAases of other human origin, whereas anti-(seminal RNAase I), -(RNAase II) and -(RNAase IV) antisera exhibited indistinguishable immunological reactions with serum RNAase and other human RNAases, except that anti-(seminal RNAase I) and -(RNAase antisera IV) did not react with pancreatic RNAases . Seminal RNAases I and IV were identical immunologically as shown by anti-(RNAase I) and anti-(RNAase IV) in immunodiffusion . Immunohistochemical study revealed that, among human tissues examined, only prostate expressed seminal RNAase III . These results suggested that human seminal RNAase I may be an aggregated molecule of RNAase IV and that seminal RNAases II and IV are similar to serum RNAases, whereas seminal RNAase III is a prostate-specific enzyme.

Sabouraudia, 1983 Dec, 21(4), 303 - 15
The use of Histolyn CYL in an enzyme immunoassay to detect Histoplasma capsulatum antibodies; Boyer MJ et al.; Histolyn CYL, the yeast phase reagent, has been employed as the antigen in an enzyme immunoassay (ELISA) to detect antibody to Histoplasma capsulatum in sera from experimentally sensitized rabbits and from humans with histoplasmosis . Three different lots of Histolyn CYL were initially evaluated with respect to antibody detection in individual and pooled rabbit serum specimens . Optimal reactivity was obtained with 4.5 micrograms (total dry weight) per ml of lots 2 and 3 of the reagent . The optimally diluted pooled reagent detected antibody titers ranging from 1:16 to 1:8192 in 36 rabbit sera . In contrast, a marked decrease in sensitivity was evidenced when the same sera were assayed by complement fixation and immunodiffusion tests with commercially available reference reagents . Complement fixation titers ranged from 0 to 1:128 and immunodiffusion tests on undiluted sera were positive with only 5 specimens . Twenty-five sera from patients with acute, chronic and disseminated histoplasmosis were also assayed by the ELISA with the reagent described above . Titers ranged from less than 1:16 to 1:20,480 or greater . Complement fixation titers ranged from 0 to 1:1024 (mycelial antigen) and from 0 to 1:256 (whole yeast cell antigen) . No antibody was detected in 7 specimens with mycelial antigen or in 3 specimens with the yeast cell antigen . Immunodiffusion tests on undiluted sera were positive with 9 of 11 specimens . Minimal cross reactivity was evidenced when Histolyn CYL was used in the ELISA to detect anti-Coccidioides immitis antibody in sera from infected animals and humans . These data encourage the continued development of this method for the serodiagnosis of human histoplasmal infection.

J Biomol Struct Dyn, 1983 Dec, 1(4), 857 - 71
Three dimensional association of double-stranded helices are produced in conditions for Z-DNA formation; Revet B et al.; The Z form of alternating poly(dG-dC).poly(dG-dC) can be induced when the concentration of NaCl, MgCl2 or ethanol are increased . In order to obtain more information concerning this Z structure, the B----Z transition is analyzed on the same sample, both by UV spectrophotometry and electron microscopy . The procedures used in this work provide high resolution images with minimal alterations of the molecules . It is shown that at high values of cations or ethanol, the polymer makes complex associations of numerous molecules stuck together parallelly . By decreasing the salt or ethanol concentrations, a progressive decondensation of the molecules is obtained . At low concentrations of Mg++ (2.10(-2) M), alterations of the linear secondary structure of the molecules are observed, although the UV spectrum is of the B-type . In the presence of that low concentration of Mg++, natural DNAs (phi X174 and yeast mitochondrial DNA fragment inserted in pBR) exhibit structural modifications similar to those observed with the poly(dG-dC).poly(dG-dC) . These structures mainly consist in four-stranded hairpins and loops built up by the sticking of two segments of DNA . The correlation between these intertwining of short DNA segments and the presence of potentially Z-forming sequences is discussed.

J Biomol Struct Dyn, 1983 Dec, 1(3), 639 - 46
Lead ion binding and RNA chain hydrolysis in phenylalanine tRNA; Rubin JR et al.; Crystalline complexes of yeast phenylalanine tRNA and Lead (II) ion were prepared by soaking pregrown orthorhombic crystals of tRNA in saturated lead chloride solutions . The locations of tightly bound lead ions on the tRNA were determined by difference Fourier methods . There are three major lead binding sites; two of these replace tightly bound magnesium ions in the native tRNA structure . Site I is located in the dihydrouridine loop of the molecule adjacent to phosphate P18 which is specifically cleaved by lead . This is evident from changes observed in the Pb-native difference electron density maps . A possible mechanism for lead ion hydrolysis of the polynucleotide chain is proposed.

Biophys J, 1983 Dec, 44(3), 353 - 63
"Peroxidatic" form of cytochrome oxidase as studied by X-ray absorption spectroscopy; Chance B et al.; X-ray absorption spectroscopy shows pulsed oxidase to be similar to resting oxidase but to lack the sulfur bridge between iron and copper of active sites (Powers, L., Y . Ching, B . Chance, and B . Muhoberac, 1982, Biophys . J., 37{2, Pt . 2}: 403a . {Abstr.} ) The first shell ligands and bond lengths of the pulsed oxidase active site heme most clearly fit the ferric peroxidases from horseradish and yeast, and the pulsed oxidase cyanide compound resembles the low spin hemoprotein cyanide compounds . The structural results are consistent with an aquo or a peroxo form for pulsed oxidase as is also observed by optical studies . These structural and chemical data are consistent with a role for the pulsed forms in a cyclic peroxidatic side reaction in which the pulsed and pulsed peroxide compounds act as peroxide scavengers . The peroxidatic role of cytochrome oxidase in the nonsulfur bridged form suggests the renaming of the "oxygenated" or "pulsed" forms on a functional basis as "peroxidatic" forms of cytochrome oxidase.

Bull Soc Pathol Exot Filiales, 1983 Dec, 76(5 Pt 2), 777 - 84
A possible case of Lôbo's disease acquired in Europe from a bottle-nosed dolphin (Tursiops truncatus); Symmers WS; A granuloma of the skin of one hand, accompanied by supratrochlear lymphadenitis, became evident about 3 months after the patient, an aquarium attendant, had had occupational contact, in Europe, with a bottle-nosed dolphin (Tursiops truncatus) . The dolphin, when caught in the Bay of Biscay, had granulomas of the skin: yeast-like organisms, morphologically indistinguishable from Loboa loboi, were found in a biopsy specimen . Identical organisms were present in the attendant's skin lesion and supratrochlear lymph node.

J Clin Invest, 1983 Dec, 72(6), 1889 - 900
Purine metabolism in myeloid precursor cells during maturation . Studies with the HL-60 cell line; Lucas DL et al.; In studies with the human promyelocytic leukemia cell line HL-60, we defined changes in intermediary purine metabolism that appear to contribute to the regulation of terminal maturation in myeloid cells . When HL-60 cells were exposed to compounds that induce maturation, consistent alterations in purine metabolism were found to occur within 24 h of culture . Perturbation of guanosine nucleotide synthesis and decreases of up to 50% in intracellular guanylate pool sizes were associated with the induced maturation of these cells in response to diverse inducing agents . While immature HL-60 cells were observed to synthesize purine nucleotides by both de novo and salvage pathways, the activity of both pathways decreased in cells induced to mature, although the relative contribution of purine salvage increased . Moreover, incorporation of the salvage pathway precursor, {14C}hypoxanthine from the intermediate, inosine monophosphate (IMP), into guanylates was reduced by approximately 65% in induced HL-60 cells, reflecting decreased activity of both hypoxanthine phosphoribosyltransferase and IMP dehydrogenase . When various inhibitors of IMP dehydrogenase (mycophenolic acid, 3-deazaguanosine, and 2-beta-D-ribofuranosylthiazole-4-carboxamide) were evaluated for their effects upon HL-60 cells, each agent was found to induce the cells to mature morphologically and functionally . Like other inducers, these agents decreased HL-60 cell proliferation and caused the cells to acquire an ability to phagocytose opsonized yeast and reduce nitroblue tetrazolium . Each agent reduced intracellular guanosine nucleotide pool sizes and induced HL-60 cell maturation at micromolar concentrations . These observations suggest that the size of intracellular guanosine nucleotide pools, the biosynthesis of guanosine nucleotides, and the activity of IMP dehydrogenase may be central to the regulation of terminal maturation in myeloid cells.

Nucleic Acids Res, 1983 Nov 25, 11(22), 8111 - 20
Nucleotide sequences of the 5.8S rRNAs of a mollusc and a porifer, and considerations regarding the secondary structure of 5.8S rRNA and its interaction with 28S rRNA; Ursi D et al.; We report the primary structures of the 5.8 S ribosomal RNAs isolated from the sponge Hymeniacidon sanguinea and the snail Arion rufus . We had previously proposed (Ursi et al., Nucl . Acids Res . 10, 3517-3530 (1982)) a secondary structure model on the basis of a comparison of twelve 5.8 S RNA sequences then known, and a matching model for the interaction of 5.8 S RNA with 26 S RNA in yeast . Here we show that the secondary structure model can be extended to the 25 sequences presently available, and that the interaction model can be extended to the binding of 5.8 S RNA to the 5'-terminal domain of 28 S (26 S) RNA in three species.

Biochem Biophys Res Commun, 1983 Nov 15, 116(3), 1107 - 13
Affinity inactivation of bovine Cu,Zn superoxide dismutase by hydroperoxide anion, HO2-; Fuchs HJ et al.; Bovine liver Cu,Zn superoxide dismutase (SOD) is inactivated by hydrogen peroxide at alkaline pH, and full inactivation correlates with the loss of 1.1 histidine/subunit . At each pH utilized, saturation of the rate of inactivation is observed . This process is characterized by a half-saturation constant for peroxide and a maximum pseudo-first-order rate constant for inactivation . At 25 degrees C, the former decreases from 15.7 to 3.2 mM as the pH is increased from 9.0 to 11.5, while the latter increases from 0.83 to 2.43 per min over the same pH range . We have previously (Arch . Biochem . Biophys . 224, 579 (1983} proposed that the true affinity reagent for the inactivation of yeast SOD is the hydroperoxide anion, and we now believe the same is true for bovine SOD . However, a subtle difference between the two enzymes exists, for while the maximum pseudo-first-order rate constant for inactivation of bovine SOD increases with increasing pH, the same parameter for the yeast enzyme is pH-independent.

J Mol Biol, 1983 Nov 5, 170(3), 777 - 90
Conservation of a DNA-binding site in the largest subunit of eukaryotic RNA polymerase II; Carroll SB et al.; Using a monoclonal antibody to a DNA-binding site of calf RNA polymerase II, we found that this site occurs on the largest subunit and is structurally similar in RNA polymerase II of widely divergent eukaryotes . In immuno-blotting of electrophoretically separated subunits, the monoclonal antibody recognized a determinant on the largest polypeptide of all RNA eukaryotic polymerase II forms tested, with a preference for the IIA enzyme subunit of 215 X 10(3) Mr over the partially proteolyzed 180 X 10(3) Mr form . This site is conserved on human, chicken, Drosophila, wheat germ and yeast RNA polymerase II, all of which reacted strongly with the monoclonal antibody . These results contrasted with those obtained with polyclonal antibodies to non-functional determinants of the calf enzyme . The reactivity of the polyclonal antibody with eukaryotic RNA polymerase II steadily decreased with increasing evolutionary distance from the original antigen; the yeast enzyme showed no cross-reactivity . These results suggest that a basic functional feature of eukaryotic RNA polymerase II has been strongly conserved and support the view that divergence of RNA polymerase II has taken place mainly in other, perhaps regulatory, sites of the enzyme.

Eur J Biochem, 1983 Nov 2, 136(2), 245 - 52
Acetylation and methylation sites in histone H4 from Physarum polycephalum; Waterborg JH et al.; Histone H4 has been isolated and purified from plasmodia of Physarum polycephalum . The four major fragments produced by hydrolysis of H4 by acetic acid were separated and the complete amino acid sequence of two of them was determined . By analogy with calf H4, these peptides are at the C-terminus and give the sequence from residue 68 to the C-terminus (residue 102) . In this 35 residue sequence there are two minor differences from calf H4: (i) residue 77 is arginine in Physarum H4 and lysine in calf H4; (ii) lysine-79 is partially methylated in Physarum . Arginine occurs at position 77 in pea H4 but the occurrence of methylated lysine at position 79 has not been reported for other species . In the N-terminal region, amino acid compositions of acetic acid, tryptic and chymotryptic peptides indicate that Physarum H4 and calf H4 have identical sequences from the N-terminus to residue 47 . There may be minor differences in the region from residue 46 to residue 67 . The sites of acetylation were determined by Edman degradation of acetate-labelled peptide 4-17 of Physarum H4 . Acetylation was observed at positions 5, 8, 12, and 16 . The only other labelled peptide was the N-terminal peptide, which is not susceptible to Edman degradation and is thus probably alpha-N-acetylated as in most other organisms . The results confirm the conservation of H4 sequence and place Physarum H4 in an intermediate position between lower eukaryote H4, such as yeast or Tetrahymena H4, and higher eukaryote H4, such as mammalian H4 or pea H4.

J Cell Biol, 1983 Nov, 97(5 Pt 1), 1476 - 90
A rat monoclonal antibody reacting specifically with the tyrosylated form of alpha-tubulin . II . Effects on cell movement, organization of microtubules, and intermediate filaments, and arrangement of Golgi elements; Wehland J et al.; A rat monoclonal antibody against yeast alpha-tubulin (clone YL 1/2; Kilmartin, J . V., B . Wright, and C . Milstein, 1982, J . Cell Biol., 93:576-582) that reacts specifically with the tyrosylated form of alpha-tubulin and readily binds to tubulin in microtubules when injected into cultured cells (see Wehland, J., M . C . Willingham, and I . V . Sandoval, 1983, J . Cell Biol., 97:1467-1475) was used to study microtubule organization and function in living cells . Depending on the concentration of YL 1/2 that was injected the following striking effects were observed: (a) When injected at a low concentration (2 mg IgG/ml in the injection solution), where microtubules were decorated without changing their distribution, intracellular movement of cell organelles (saltatory movement) and cell translocation were not affected . Intermediate concentrations (6 mg IgG/ml) that induced bundling but no perinuclear aggregation of microtubules abolished saltatory movement and cell translocation, and high concentrations (greater than 12 mg IgG/ml) that induced perinuclear aggregation of microtubules showed the same effect . (b) YL 1/2, when injected at intermediate and high concentrations, arrested cells in mitosis . Such cells showed no normal spindle structures . (c) Injection of an intermediate concentration of YL 1/2 that stopped saltatory movement caused little or no aggregation of intermediate filaments and no dispersion of the Golgi complex . After injection of high concentrations, resulting in perinuclear aggregation of microtubules, intermediate filaments formed perinuclear bundles and the Golgi complex became dispersed analogous to results obtained after treatment of cells with colcemid . (d) When rhodamine-conjugated YL 1/2 was injected at concentrations that stopped saltatory movement and arrested cells in mitosis, microtubule structures could be visualized and followed for several hours in living cells by video image intensification microscopy . They showed little or no change in distribution and organization during observation, even though these microtubule structures appeared not to be stabilized by injected YL 1/2 since they were readily depolymerized by colcemid or cold treatment and repolymerized upon drug removal or rewarming to 37 degrees C, respectively . These results are discussed in terms of the participation of microtubules in cellular activities such as cell movement and cytoplasmic organization and in terms of the specificity of YL 1/2 for the tyrosylated form of alpha-tubulin.

Zh Mikrobiol Epidemiol Immunobiol, 1983 Nov, (11), 41 - 5
{Isolation of the 1st strains of Legionella pneumophila in the USSR}; Tartakovskii IS et al.; For the first time L . pneumophila strains were isolated from pneumonia patients in the USSR . To isolate these strains, the material obtained from the patients was inoculated into charcoal-yeast agar with antibiotics or into guinea pigs with the subsequent inoculation of chick embryos . Both isolated strains were classified with serogroup 1 of L . pneumophila on the basis of their cultural, morphological, biochemical and serological properties.

Tohoku J Exp Med, 1983 Nov, 141(3), 295 - 303
Monocyte function in idiopathic nephrotic syndrome in childhood; Waga S et al.; The monocyte function of 112 specimens from 42 children with idiopathic nephrotic syndrome (INS) aged from 2 to 17 years was studied by the methods of nitroblue tetrazolium (NBT) reduction, phagocytosis of immunobeads (IB) and yeast cells, chemotaxis and acid alpha-naphthyl acetate esterase (ANAE) staining . The dissociation between phagocytosis and chemotaxis was observed in the fresh cases of steroid sensitive INS and in the cases of steroid non-sensitive INS . In the fresh cases of steroid sensitive INS, NBT reduction and phagocytosis were increased, but chemotaxis was decreased . In the cases of steroid non-sensitive INS, the phagocytosis of IB was decreased, but chemotaxis was increased . These findings suggest a different pathogenesis between steroid sensitive and steroid non-sensitive INS . The dissociation between phagocytosis and chemotaxis may be explained by the alteration of the surface receptors of monocyte and by lymphokines.

J Nutr, 1983 Nov, 113(11), 2147 - 54
Factors influencing the antitumorigenic properties of selenium in mice; Poirier KA et al.; Sodium selenite (Na2SeO3) was administered at 2.0 micrograms selenium (Se) to Swiss ICR mice six times over a 9-day period by intraperitoneal (i.p.) injection or by gastric gavage . Survival time was significantly increased in Ehrlich ascites tumor (EAT)-bearing mice by 170 and 20%, respectively, compared to controls . In two separate studies, 5.0 micrograms Se as Na2SeO3 or selenodiglutathione (GSSeSG) administered i.p . was more effective in inhibiting EAT propagation in mice than either untreated (control) mice or mice receiving sodium selenide, dimethyl selenide {(CH3)2Se} or seleno-DL-cystine . In another study, EAT cells were preincubated with either 1 or 3 ppm Se as GSSeSG, Na2SeO3, or (CH3)2Se, washed, and reinoculated into mice . Only in mice inoculated with cells pretreated with GSSeSG was a significant increase in survival observed . The observed tumor inhibition was not limited to ascitic tumors since growth of solid Ehrlich tumors was also significantly inhibited by i.p . treatment of Na2SeO3 . Following i.p . administration of Na2(75)SeO3, the solid tumors retained more selenium-75 than did blood, lung, kidney, or liver . Supplementation of a torula yeast diet with 2.5 or 5.0 ppm Se as Na2SeO3 also significantly increased the survival time of EAT-bearing mice . These data show that the form and mode of administration of selenium influence the antitumorigenic properties of this trace element . In addition, the data suggest that some intermediate in the normal pathway for selenium detoxification is probably responsible for this trace element's antitumorigenic properties.

Arch Pathol Lab Med, 1983 Nov, 107(11), 577 - 9
Pathogenicity of Candida paratropicalis; Baker JG et al.; A new Candida species, Candida paratropicalis, was recently described . Four cases of infections due to C paratropicalis are reviewed in detail and an additional five cases are reviewed to establish the clinical relevance of this species of yeast . Candida paratropicalis was isolated from blood and several other body sites . Although the isolates tested were sensitive in vitro to amphotericin B and fluocytosine, significant morbidity and mortality were associated with the infections.

Can J Biochem Cell Biol, 1983 Nov, 61(11), 1201 - 7
Thermodynamic profiles for alcohol dehydrogenase action in free solution; Laidler KJ et al.; Stopped-flow equipment was used to study the kinetics of the reaction between nicotinamide adenine dinucleotide (NAD) and ethanol, catalyzed by yeast alcohol dehydrogenase . By measuring rates over a range of concentrations of NAD and ethanol and of temperatures, thermodynamic profiles were obtained for the reaction, which occurs by an ordered ternary complex mechanism with NAD adding first . There are significant negative entropies of activation and negative entropy changes for the addition of NAD and of ethanol; the breakdown of the ternary complex is, however, accompanied by a positive entropy of activation . The results are consistent with structural constraints associated with the binding of the substrates, these restraints being to some extent removed when the ternary complex undergoes reaction . The system follows a similar pattern to that found with three different varieties of lactate dehydrogenase.

Infect Immun, 1983 Nov, 42(2), 818 - 23
Protective effect of poly-2-vinylpyridine-N-oxide on susceptibility of silica-treated mice to experimental histoplasmosis; Von Behren LA et al.; We have studied the ability of poly-2-vinylpyridine-N-oxide (PVNO), a lysosomal stabilizing agent, to abrogate the cytotoxic effects of silica on macrophages . Male C3H/HeN mice were pretreated with PVNO and inoculated intravenously with silica particles . At 24 h after silica injection, silica-treated and -untreated mice were challenged intravenously with varying doses of live yeast cells of Histoplasma capsulatum . All mice receiving silica died when challenged with 5 X 10(5) yeast cells of Histoplasma sp . compared with no deaths in PVNO-pretreated animals and 10% mortality in controls not receiving PVNO or silica . When animals were given 2.5 X 10(5) yeast cells (a sublethal dose), the protective effect of PVNO was seen by a reduction in splenomegaly and viable Histoplasma sp . present in the spleen . Furthermore, PVNO alone showed a significant protective effect (P less than 0.05) against a lethal challenge with Histoplasma sp . Prior treatment with PVNO also protected mouse peritoneal macrophages from the cytotoxic effects of silica particles in vitro . These results indicate that PVNO abrogates the cytotoxicity of silica particles on macrophages and also increases the resistance of mice to histoplasmosis.

J Immunol, 1983 Nov, 131(5), 2368 - 73
Lymphokines secreted by an established lymphocyte line modulate receptor-mediated endocytosis in macrophages derived from human monocytes; Fogelman AM et al.; As little as 1 microliter of serum-free supernatant from Mo(t), an established lymphocyte line, when added to a 500-microliters incubation of macrophages derived from human monocytes, significantly decreased the receptor-mediated uptake and degradation of three cholesterol-rich molecules: low density lipoprotein (LDL); LDL complexed to dextran sulfate; and LDL modified by malondialdehyde (MDA-LDL) . In contrast, the receptor-mediated uptake and degradation of mannosyl bovine serum albumin was increased three-fold . The Mo(t) supernatant did not contain competitive inhibitors of the cholesterol-rich ligands, and it did not alter macrophage receptor-independent endocytosis, protein synthesis, or phagocytosis of heat-killed yeast . The effect of the Mo(t) supernatant was specific for macrophages and was abolished by preincubation of the supernatant with trypsin, which indicates that the active substances are protein in nature . The decrease in the uptake and degradation of MDA-LDL induced by preincubating the macrophages with Mo(t) supernatant appeared to result from a decrease in the number of receptors for this ligand at the cell surface . The isolation of these lymphokines should offer new insights into macrophage receptor-mediated endocytosis, and may yield substances useful in preventing foam cell formation in atherosclerosis.

J Clin Microbiol, 1983 Nov, 18(5), 1252 - 5
Candida ciferrii and Candida chiropterorum isolated from clinical specimens; Furman RM et al.; Ten clinical yeast isolates submitted to the Centers for Disease Control from diverse geographic areas were identified as Candida ciferrii and Candida chiropterorum . The association of C . ciferrii with clinical specimens, particularly its repeated isolation from a case of onychomycosis, suggests that this species may be an etiological agent of superficial yeast infections.

Biochem Biophys Res Commun, 1983 Oct 31, 116(2), 383 - 7
Rat liver protoporphyrinogen IX oxidase: site of synthesis and factor influencing its activity; Kolarov J et al.; Rat liver protoporphyrinogen IX oxidase is not formed in mitochondria in contrast to the claims made for the yeast enzyme (Poulson and Polglase, FEBS Lett . (1974) 40, 258) . Inhibition of mitochondrial protein synthesis in regenerating rat livers by thiamphenicol led, instead, to a slight increase in protoporphyrinogen oxidase activity . Protoporphyrinogen IX oxidase was not induced in rat liver by triiodothyronine, an inducer of mitochondrial protein synthesis, or by AIA, an inducer of heme synthesis . Significant increases in activity were observed to be associated with rapidly growing cells, such as regenerating livers and rat ascites hepatoma cells.

J Biol Chem, 1983 Oct 25, 258(20), 12386 - 93
Phaseolus vulgaris cytoplasmic leucyl-tRNA synthetase . Purification and comparison of its catalytic, structural, and immunological properties with those of the chloroplastic enzyme; Dietrich A et al.; The cytoplasmic leucyl-tRNA synthetase was purified from bean (Phaseolus vulgaris) leaves . After ammonium sulfate fractionation and chromatography on Sephadex G-50, DEAE-cellulose, hydroxylapatite, and phosphocellulose, complete purification was achieved by blue Sepharose CL-6B chromatography using specific elution with pure yeast tRNALeu1 . The enzyme was purified 1050-fold and had a specific activity of 940 nmol of leucyl-tRNA formed/min/mg of protein . Polyacrylamide gel electrophoresis of the native enzyme showed one band, but the denatured enzyme showed two bands . These two protein bands are structurally related . The smallest protein appears to be a cleavage product from the largest one, suggesting the presence of a sensitive cleavage site in the cytoplasmic leucyl-tRNA synthetase . The cytoplasmic enzyme is a monomer (Mr = 130,000), larger than its chloroplastic counterpart (Mr = 120,000) . The two enzymes differ in their substrate (tRNA) specificity, tryptic peptide map, and amino acid composition . Antibodies were raised against the cytoplasmic enzyme and against the chloroplastic enzyme and no cross-immunological reaction was detected, showing that the two enzymes do not share any antigenic determinant . Taken together, these results suggest that P . vulgaris cytoplasmic and chloroplastic leucyl-tRNA synthetases are coded for by different genes.

Nucleic Acids Res, 1983 Oct 11, 11(19), 6859 - 72
Genes for cytochrome c oxidase subunit I, URF2, and three tRNAs in Drosophila mitochondrial DNA; Clary DO et al.; Genes for URF2, tRNAtrp, tRNAcys, tRNAtyr and cytochrome c oxidase subunit I (COI) have been identified within a sequenced segment of the Drosophila yakuba mtDNA molecule . The five genes are arranged in the order given . Transcription of the tRNAcys and tRNAtyr genes is in the same direction as replication, while transcription of the URF2, tRNAtrp and COI genes is in the opposite direction . A similar arrangement of these genes is found in mammalian mtDNA except that in the latter, the tRNAala and tRNAasn genes are located between the tRNAtrp and tRNAcys genes . Also, a sequence found between the tRNAasn and tRNAcys genes in mammalian mtDNA, which is associated with the initiation of second strand DNA synthesis, is not found in this region of the D . yakuba mtDNA molecule . As the D . yakuba COI gene lacks a standard translation initiation codon, we consider the possibility that the quadruplet ATAA may serve this function . As in other D . yakuba mitochondrial polypeptide genes, AGA codons in the URF2 and COI genes do not correspond in position to arginine-specifying codons in the equivalent genes of mouse and yeast mtDNAs, but do most frequently correspond to serine-specifying codons.

J Biol Chem, 1983 Oct 10, 258(19), 11974 - 80
Characterization of tRNA precursor splicing in mammalian extracts; Laski FA et al.; Transcription of a Xenopus laevis tRNATyr gene and splicing of the transcript have been studied in HeLa cell extracts . This tRNATyr gene has a 13-base intervening sequence and is expressed as mature tRNA when transfected into mammalian cells . The tRNATyr gene is transcribed under conditions of low concentrations of magnesium and ATP, but is processed by splicing only when both of these cofactors are added at higher concentrations . The endonucleolytic activity of the tRNA-splicing system in the HeLa extract produces exons with 3'-phosphate and 5'-hydroxyl groups . The 3'-phosphate is retained during the ligation reaction and forms the phosphodiester bond in the mature tRNA . Retention of the 3'-phosphate during tRNA splicing differs from the more extensively studied process in yeast extracts where a phosphate group from an ATP cofactor is used to form the phosphodiester bond joining the exons . Thus, eucaryotic organisms can splice tRNA precursors by at least two distinguishable mechanisms.

Can J Microbiol, 1983 Oct, 29(10), 1253 - 7
Metabolism of cinnamic, p-coumaric, and ferulic acids by Streptomyces setonii; Sutherland JB et al.; Streptomyces setonii strain 75Vi2 was grown at 45 degrees C in liquid media containing yeast extract and trans-cinnamic acid, p-coumaric acid, ferulic acid, or vanillin . Gas chromatography, thin-layer chromatography, and mass spectrometry showed that cinnamic acid was catabolized via benzaldehyde, benzoic acid, and catechol; p-coumaric acid was catabolized via p-hydroxybenzaldehyde, p-hydroxybenzoic acid, and protocatechuic acid; ferulic acid was catabolized via vanillin, vanillic acid, and protocatechuic acid . When vanillin was used as the initial growth substrate, it was catabolized via vanillic acid, guaiacol, and catechol . The inducible ring-cleavage dioxygenases catechol 1,2-dioxygenase and protocatechuate 3,4-dioxygenase were detected with an oxygen electrode in cell-free extracts of cultures grown in media with aromatic growth substrates and yeast extract.

Arch Dis Child, 1983 Oct, 58(10), 799 - 802
A common congenital immunodeficiency predisposing to infection and atopy in infancy; Richardson VF et al.; Twenty six infants with a congenital immunodeficiency, characterised by failure of their sera to opsonise heat killed bakers' yeast for phagocytosis by normal polymorphonuclear leucocytes, were studied during infancy to determine the frequency of infection and development of atopy . They were compared with controls, matched prospectively for birth date, sex, parental smoking, and atopy and in whom feeding patterns were similar . In 18 of 26 infants the serum defect persisted at age one year . The incidence of infection and atopy, was appreciably greater in the study group than in controls . The 8 children in whom the defect was transient had a similar incidence of infection but a higher incidence of atopy than controls . Eight of 26 mothers and four of 9 fathers tested also had the serum defect, suggesting a strong genetic component . We support the hypothesis that immunodeficiency predisposes to infection and atopy, and that transient immunodeficiency predisposes to atopy.

Proc Soc Exp Biol Med, 1983 Oct, 174(1), 12 - 5
Culture of human gastrointestinal epithelial cells; Moyer MP; Human gastrointestinal (GI) epithelial cells and tissues have been propagated in vitro for several months . Viability of typical epithelial cells with minimal contamination by fibroblasts was best achieved by mechanical harvesting instead of using enzymes for dissociation or subculture . Individual cells generally adhered poorly to glass and plastic although they did attach to some coated substrates . Although standard formulations of culture base media were suitable, there was a continuous requirement for conditioned medium and cultures were readily propagated in suspension . Cultures had a propensity to grow as "islands" of epithelial cells consisting of central dividing regions and external "differentiated" regions . As yet undefined factors from yeast and brain (or pituitary) extracts were required for optimal growth.

J Bacteriol, 1983 Oct, 156(1), 447 - 9
Control of triglyceride synthesis by the intracellular level of long-chain acyl coenzyme A for lipid synthesis; Kamiryo T; Candida lipolytica mutants defective in acyl coenzyme A synthetase I synthesized triglyceride to a markedly less extent than did the wild-type yeast, when grown on oleic acid . The synthesis of triglyceride was controlled by the level of long-chain acyl coenzyme A available for lipid synthesis, whereas the synthesis of phospholipids was hardly affected.

Am J Nurs, 1983 Oct, 83(10), 1392 - 8
The pill: a closer look; Dickerson J; PIP: This article summarizes the major risks and benefits of oral contraceptive (OC) use for specific categories of users . Major risks associated with OC use include vascular and circulatory disorders, hypertension, cancer, and other conditions such as gallbladder disease . There are also numerous minor side effects, e.g., breast tenderness, weight changes, yeast infections . Most of these side effects are attributed either to estrogen or progestin, which mimic excesses or deficiencies in the natural hormonal balance . These symptoms can often be reversed through alterations in the hormonal content of the OC . There have been numerous recent reports regarding the protective effect of OC use against conditions such as benign breast disease, ovarian and endometrial cancer, pelvic inflammatory disease, ectopic pregnancy, and rheumatoid disease . The risks and benefits for potential users can only be evaluated through reference to data from the relevant population group, taking into account factors such as age, race, heredity, potential predisposition for disease, and social habits . Information about the risk of medical problems in specific population groups must be weighed against the risk for those problems in the same population when combined with OC treatment . The benefits of the drug must also be weighed against the number and degree of risks found for the specific user . The convenience and efficacy associated with OCs can far outweigh the risks, inconveniences, and less impressive efficacy of other contraceptive methods in many cases . However, women over age 35 years, especially smokers, should use alternative methods of contraception . New hormonal contraceptive formulations and different modes of drug delivery are currently under development . However, several years of scientific investigation will be required to evaluate the longterm advantages or disadvantages of the newer experimental drugs compared with present OCs .

J Cell Biol, 1983 Oct, 97(4), 1011 - 9
Identification of tubulin in Dictyostelium discoideum: characterization of some unique properties; White E et al.; We used three antitubulin antibodies to localize Dictyostelium tubulin subunits on two-dimensional polyacrylamide gels by Western blotting . All three antibodies, a polyclonal antibody against sea urchin alpha- and beta-tubulin and two monoclonal antibodies against yeast alpha-tubulin, recognize the same set of polypeptides with a molecular weight of 55,000 while focusing at a pH far more basic than all other tubulins . Each antibody specifically stains the microtubule system of slime mold amoebae by indirect immunofluorescence . The microtubule system can be isolated as a major component of the amoeba cytoskeleton, and these preparations are greatly enriched for the presumptive tubulin subunits . The microtubules of these cytoskeletons are resistant to being depolymerized by millimolar concentrations of calcium, while they retain their cold sensitivity . Comparison of peptide maps of slime mold and brain alpha-tubulins indicates that the proteins are related but not identical . Possible explanations for these unusual characteristics are discussed.

Am J Physiol, 1983 Oct, 245(4), R478 - 83
Temporal self-organization in biochemical systems: periodic behavior vs . chaos; Goldbeter A et al.; The patterns of temporal self-organization in regulated biochemical systems are examined . Simple periodic oscillations are the most frequent type of such organization, as exemplified by glycolytic oscillations in yeast and muscle and by the periodic synthesis of adenosine 3',5'-cyclic monophosphate in Dictyostelium discoideum amoebas . These phenomena originate, respectively, from the periodic operation of the product-activated phosphofructokinase and adenylate cyclase reactions . The analysis of a model for a multiply regulated biochemical system shows more complex oscillatory phenomena, e.g., the coexistence between two stable periodic regimes for the same set of parameter values (birhythmicity) and chaos . The latter phenomenon of aperiodic oscillations occurs in a narrow range of parameter values and is much less frequent than simple or complex periodic behavior . It is suggested that a sufficient condition for the occurrence of birhythmicity and chaos in a regulated biological system subjected to a constant environment (i.e., in the absence of periodic forcing) may be the simultaneous presence and interaction of two mechanisms capable of producing oscillations.

Mol Cell Biol, 1983 Oct, 3(10), 1694 - 702
Sequence analysis of mitochondrial DNA in a mouse cell line resistant to chloramphenicol and oligomycin; Slott EF Jr et al.; A mouse L-cell line, designated 111-OB3, is described which is resistant to two drugs, chloramphenicol and oligomycin . The cells contain two types of mitochondrial DNA molecules, in roughly equal proportions, which differ in that one is cleaved by endonuclease EcoRI at a novel site within the coding sequence for subunit 6 of the mitochondrial ATPase (ATPase-6) . Sequence analysis reveals that the cleavage site was created by a single transversion which predicts a replacement of valine in the wild-type ATPase-6 by glutamic acid . The replacement occurs in a hydrophobic amino acid sequence which is highly conserved in mouse, human, and bovine proteins . The position of the replacement is similar to a substitution observed in one class of yeast mutants resistant to oligomycin . Both of the mitochondrial DNA molecules in 111-OB3 also have a single nucleotide change in the gene encoding the large (16S) rRNA . These observations are consistent with the hypothesis that oligomycin resistance in mammalian cells can be cytoplasmically determined and can result from alterations in ATPase-6 . The appearance of the mutation before selection in oligomycin suggests a model for the origin of mitochondrial mutations in mammalian cells.

Biochem Biophys Res Commun, 1983 Sep 30, 115(3), 1101 - 7
Difference in hydrophobicity between mitochondria-bindable and non-bindable forms of hexokinase purified from rat brain; Kurokawa M et al.; Hexokinase able to bind to mitochondria was purified to homogeneity from rat brain by two successive DEAE-cellulose chromatographic steps . The enzyme lost only the binding ability with almost undetectable change in molecular weight on mild chymotrypsin digestion . The bindable hexokinase was adsorbed to a Phenyl-Sepharose column and eluted with a Lubrol PX gradient, whereas non-bindable hexokinase and yeast hexokinase were not adsorbed under the similar conditions . These results suggest that mitochondria-bindable hexokinase has a hydrophobic region on its surface, which is responsible for the specific interaction with mitochondria.

Biochem Biophys Res Commun, 1983 Sep 30, 115(3), 971 - 80
Induced maturation of the human promyelocytic leukemia cell line, HL-60, by 2-beta-D-ribofuranosylselenazole-4-carboxamide; Lucas DL et al.; The new synthetic nucleoside analogue, 2-beta-D-ribofuranosylselenazole-4-carboxamide, was evaluated for its effects upon the growth and maturation of the human promyelocytic leukemia cell line, HL-60 . At a concentration of greater than or equal to 1 nm, this agent was found both to decrease HL-60 cell proliferation and to cause the cells to acquire an ability to phagocytose opsonized yeast and to reduce nitroblue tetrazolium dye, functions characteristic of mature myeloid cells . In addition, this agent at similar concentrations caused a marked depression of intracellular guanosine nucleotide pools and a reduction in the incorporation of {14C} hypoxanthine into guanylates . These results suggested that the selenazole nucleoside caused an inhibition of inosinate monophosphate dehydrogenase, a key enzyme of guanylate biosynthesis . We therefore measured the activity of this enzyme indirectly by simultaneous-UV-radioactivity HPLC as well as by a direct radiometric method and demonstrated markedly reduced enzyme activities by both assays in drug treated cells . Dose response studies indicated that concentrations of drug which caused greater than 30% inhibition of IMP dehydrogenase activity induced greater than 50% maturation of the cells . These observations with this new nucleoside analogue provide further support for the concept that production of guanosine nucleotides and the activity of IMP dehydrogenase have a role in regulating the terminal maturation of myeloid cells.

J Biol Chem, 1983 Sep 25, 258(18), 11014 - 9
The reconstitution of denatured phosphoglycerate mutase; Hermann R et al.; The reconstitution of the tetrameric enzyme yeast phosphoglycerate mutase after denaturation in guanidine hydrochloride has been studied . Denaturation is almost completely reversible at enzyme concentrations greater than 10 micrograms/ml . Cross-linking by glutaraldehyde has been used to monitor the reassociation of the subunits; the kinetics of this process has been analyzed in terms of a model involving an equilibrium between monomer and dimer followed by a bimolecular association of two dimers to give a tetramer . Reactivation is found to parallel the appearance of tetramer . Structural changes during reconstitution have been measured by circular dichroism and fluorescence . Both methods reveal complex kinetics indicating the rapid formation of structured monomers (half-time less than 10 s), followed by slow subunit association . For comparison, preliminary reconstitution experiments were performed on the dimeric phosphoglycerate mutase from rabbit muscle.

J Bacteriol, 1983 Sep, 155(3), 1088 - 93
Two distinct classes of polyuronide from the cell walls of a dimorphic fungus, Mucor rouxii; Dow JM et al.; Polyuronides were extracted from purified yeast and mycelial walls of Mucor rouxii by sequential treatments with lithium chloride and potassium hydroxide and were fractionated by ion-exchange chromatography on DEAE-Sephadex . Two polymers (I and II) of different acidity were found in both wall types . Polymer I contained D-glucuronic acid, L-fucose, D-mannose, and much smaller amounts of D-galactose . Yeast and mycelial polymer I had similar uronic acid contents but differed in their neutral sugar compositions and molecular weights . Polymer II from both cell types contained largely D-glucuronic acid and had similar molecular weights . On partial acid hydrolysis, both polymers I and II gave rise to insoluble glucuronans which appeared to be homopolymeric . One-third of the total uronosyl residues of polymer I, and almost all of the uronosyl residues of polymer II, were present in homopolymeric segments . However, homopolymers derived from polymers I and II may not be identical.

Infect Immun, 1983 Sep, 41(3), 908 - 12
Susceptibility of Blastomyces dermatitidis strains to products of oxidative metabolism; Sugar AM et al.; Three strains of Blastomyces dermatitidis which differ in their virulence for mice were exposed in their yeast form to various components of the peroxidase-hydrogen peroxide-halide system . Susceptibility to H2O2 alone correlated with virulence, with the most virulent strain (ATCC 26199) least susceptible (50% lethal dose, greater than 50 mM) and an avirulent strain (ATCC 26197) most susceptible (50% lethal dose less than 3.3 mM) . A strain of intermediate virulence (ATCC 26198) was of intermediate susceptibility (50% lethal dose, 11.5 mM) . The addition of a nontoxic concentration of KI (5 X 10(-4) M) did not increase H2O2 toxicity . However, the addition of either myeloperoxidase or horseradish peroxidase and KI markedly decreased the amount of H2O2 required to kill the organisms, with 100 +/- 0% of all strains killed at 5 X 10(-5) M H2O2 and 97 +/- 4, 100 +/- 0, and 94 +/- 8% of ATCC 26199, ATCC 26198, and ATCC 26197 killed, respectively, at 5 X 10(-6) M H2O2 . Kinetic studies with H2O2 alone revealed a delayed onset of killing, but virtually 100% of organisms were killed by 120 min of exposure in all strains . By comparison, the peroxidase-hydrogen peroxide-halide system was 100% lethal for all strains at 1 min . The relatively high concentrations of H2O2 required to kill the yeast phase of B . dermatitidis suggest that H2O2 alone does not account for host resistance to the organism . However, the rapidly lethal effect of the peroxidase-hydrogen peroxide-halide system at physiologically relevant concentrations suggests that this may be one mechanism of host defense to B . dermatitidis.

Zentralbl Bakteriol Mikrobiol Hyg {B}, 1983 Sep, 177(6), 499 - 506
Isolation of sporothrix schenckii from the floor of an indoor swimming pool; Staib F et al.; In a routine mycological examination of an indoor swimming pool during the winter season (January 1981) in Berlin (West), Sporothrix schenckii was isolated from the floor near a shower . The strain was characterized by cultural, morphological and biochemical properties typical of strains which are causative agents of sporotrichosis . It is suggested that in routine mycological examinations of swimming pools attention should not be limited to dermatophytes and yeast-like fungi.

Clin Allergy, 1983 Sep, 13(5), 419 - 26
Immunological studies in asthmatic children undergoing antigen provocation in the skin, lung and nose; Price JF et al.; In children with perennial asthma, dual (immediate and late) reactions in the skin to D . pteronyssinus and Timothy grass pollen were more frequent with high doses of antigen and were associated with large immediate reactions . The frequency of dual bronchial or nasal reactions was not related to the size of the immediate reaction and dual reactions were commonly elicited to the lowest antigen dose which would elicit an immediate reaction . Serum IgG, IgA, IgM or IgE concentrations, IgE or IgG antibodies to the antigens and defective yeast opsonization did not differ in children with dual or immediate only reactions in skin, nose or lung . In five patients undergoing bronchial provocation tests with D . pteronyssinus there was no fever, no fine crepitations in the lungs and no significant change in the levels of C3.

Sabouraudia, 1983 Sep, 21(3), 215 - 21
Blastomyces dermatitidis in India: first report of its isolation from clinical material; Randhawa HS et al.; The isolation of Blastomyces dermatitidis in India is reported from the bronchial aspirate of a female asthmatic patient who had never travelled abroad . The patient was a resident of Rodhain, a small town in the District of Badaun (Uttar Pradesh), situated about 250 km south-east of Delhi . Apart from its demonstration by culture and direct microscopy of a bronchial aspirate, B . dermatitidis was seen microscopically on two occasions in KOH wet mounts and smears of sputum stained with periodic acid-Schiff's reagent . Anti-B . dermatitidis serum precipitins were shown by immunodiffusion in 3 of 4 serum samples from the patient . The identity of the fungus was based on its characteristic morphology in clinical specimens and in culture, conversion of the mold form to the yeast from in vitro and vice versa, and by verification of its pathogenicity in white mice . A detailed clinical and laboratory evaluation of the patient indicated that she had suffered from an episode of self-limited acute pulmonary blastomycosis that required no antifungal therapy . This is believed to be the first authentic report of the isolation of B . dermatitidis from clinical material in India.

J Clin Psychopharmacol, 1983 Aug, 3(4), 249 - 52
Monoamine oxidase inhibitors: reappraisal of dietary considerations; Folks DG; Dangers of adverse effects with MAOIs have been exaggerated, the most serious being hypertensive interactions with cheeses, yeast extracts, certain alcoholic beverages, broad bean pods, certain meats, and foods that are unfresh or overripe . Some adverse reactions involving MAOI-diet interactions were likely due to migraine headache, consumption of unusually large quantities of low-tyramine-containing foods, or other individual variations . Since MAOIs have advantages in pharmacotherapy of certain depressive subtypes and anxiety states, knowledge of the dietary considerations with MAOI treatment is essential to both the clinician and their patients . Patients are easily educated about MAOIs and the necessary dietary restrictions using the steps outlined in this report . Finally, written information concerning the MAOI diet together with Medic-Alert emblems or drug manufacturers' cards warning of MAOI-drug interactions should be provided before MAOI treatment is begun . Although MAOI-drug interactions were not addressed in this article, in general, it is good policy for the patient to contact the clinician whenever another medication (over-the-counter or prescribed by another physician) is at issue so that its safety can be verified.

J Cell Biol, 1983 Aug, 97(2), 566 - 70
Nonlysosomal vesicles (acidosomes) are involved in phagosome acidification in Paramecium; Allen RD et al.; Although acidification of phagocytic vacuoles has received a broadened interest with the development of pH-sensitive fluorescent probes to follow the pH changes of vacuoles and acidic vesicles in living cells, the mechanism responsible for the acidification of such vacuoles still remains in doubt . In previous studies of the digestive vacuole system in the ciliate Paramecium caudatum we observed and described a unique population of apparently nonlysosomal vesicles that quickly fused with the newly released vacuole before the vacuole became acid and before lysosomes fused with the vacuole . In this paper we report the following: (a) these vesicles, named acidosomes, are devoid of acid phosphatase; (b) these vesicles accumulate neutral red as well as acridine orange, two observations that demonstrate their acid content; (c) cytochalasin B given 15 s after exposure of the cells to indicator dye-stained yeast will inhibit the acidification of yeast-containing vacuoles; and that (d) we observed using electron microscopy, that fusion of acidosomes with the vacuole is inhibited by cytochalasin B . We conclude that the mechanism for acidification of phagocytic vacuoles in Paramecium resides, at least partially if not entirely, in the acidosomes.

Lab Invest, 1983 Aug, 49(2), 148 - 53
Roles of selenium and sulfur-containing amino acids in protection against oxygen toxicity; Forman HJ et al.; Tolerance and adaptation to hyperoxia have been correlated with increases in antioxidant enzymes . This study evaluated whether selenium deficiency would prevent an increase in glutathione peroxidase (GSHPX), a selenium-containing enzyme, during oxygen exposure, and, thus, inhibit adaptation . Because the Torula yeast-based diet, which was used to produce selenium deficiency, was also deficient in cysteine and methionine, the effects of these deficiencies were also evaluated . When rats were exposed to 80% oxygen for 1 week, mortality was 80% for rats deficient in both selenium and the sulfur-containing amino acids, 40% for selenium-deficient rats, 35% for cysteine- and methionine-deficient rats, and 0% for rats fed either a standard laboratory diet or a selenium, cysteine-, and methionine-supplemented Torula yeast diet . However, only one of the six surviving rats with low selenium and none of the rats from any other dietary group died during a subsequent 96 hours of 98% oxygen, indicating adaptation to hyperoxia (LD50 for unadapted rats is 72 hours.) GSHPX activity (per gram of dry weight) was decreased 85% in lungs from unexposed rats fed the low selenium diets . After oxygen exposure, lung GSHPX activity was elevated in all dietary groups . Rats fed the high selenium diets had a 47% increase in enzyme activity, whereas rats with high selenium had a 214% increase . Although hyperoxia caused a relatively high percentage increase in the low Se rats, the resulting absolute GSHPX activity was only 34 to 70% of that of unexposed high selenium rats . The results indicate that both selenium and sulfur-containing amino acids contribute to antioxidant defense . However, although the stress of hyperoxic exposure produces an increase in glutathione peroxidase activity, the absolute lung GSHPX activity is better correlated with tolerance than with adaptation to hyperoxia.

J Clin Microbiol, 1983 Aug, 18(2), 443 - 4
Fatal septicemia due to amphotericin B-resistant Candida lusitaniae; Guinet R et al.; Five yeast strains, causally associated with septicemia and death in a patient after peritonitis, were identified as Candida lusitaniae van Uden et do Carmo-Sousa by standard methods . The organism was initially susceptible to 5-fluorocytosine but strongly resistant to amphotericin B, requiring 50 micrograms/ml for complete inhibition at 48 h.

Appl Environ Microbiol, 1983 Aug, 46(2), 400 - 5
Production of aflatoxin by an Aspergillus flavus isolate cultured under a limited oxygen supply; Clevstrom G et al.; In a previous experiment on the preservation of hay of high moisture content with formic acid, among other agents, aflatoxin was formed in the hay, and aflatoxin-forming strains of Aspergillus flavus were isolated from this hay after incubation in air as well as in an anaerobic jar . One isolate from the anaerobic jar was cultivated in a chemostat (Bioflo model C 30; New Brunswick Scientific Co.) in a defined medium with added B vitamins, yeast extract, or formic acid, with or without gas flow (air or nitrogen) . In all cases where spore germination occurred, aflatoxin was formed in the cultures with gas flow, and small quantities of aflatoxins B1 and B2 occurred even in an atmosphere of nitrogen . Addition of B vitamins and supply of traces of air gave an approximately 15-fold increase in the amount of aflatoxin in 2 days . Carbon dioxide enrichment hindered aflatoxin formation on the defined medium even in the presence of B vitamins, but when formic acid was added, small quantities (5 to 15 micrograms/liter) were formed, and this low level remained constant until the gas flow was started.

J Reticuloendothel Soc, 1983 Aug, 34(2), 99 - 111
Effect of silica on the susceptibility of mice to experimental histoplasmosis; Von Behren LA et al.; The role of macrophages in the innate immunity of mice to histoplasmosis was investigated using silica, which selectively inactivates macrophages . Mice given silica IV 1 day prior to challenge with live yeast cells of Histoplasma capsulatum were more susceptible to infection than were untreated controls . This increased susceptibility to Histoplasma was observed when mice were given silica at 1, 14, and 21 days prior to infection but not at 3 and 7 days . Silica treated mice that survived 30 days after challenge with a sublethal dose of Histoplasma had 23 times more viable organisms in their spleens than in those of untreated controls . The blastogenic response of spleen cells to concanavalin A and phytohemagglutinin was unaffected at 12 hr after silica injection but was significantly depressed between 1 and 21 days . In contrast, silica treatment did not affect the blastogenic response of spleen cells to lipopolysaccharide . Silica particles were cytotoxic for mouse peritoneal macrophages but not to lymphocytes in vitro . These results indicate that macrophages play an essential role in natural immunity to histoplasmosis.

J Immunol, 1983 Aug, 131(2), 984 - 90
Immunoregulation in experimental disseminated histoplasmosis: flow microfluorometry (FMF) studies of the Thy and Lyt phenotypes of T lymphocytes from infected mice; Watson SR et al.; Previous studies have shown that mice infected i.v . with 6 X 10(5) yeast phase Histoplasma capsulatum (Hc) develop suppressed immune responses during weeks 1 to 4 of infection but that by weeks 8 to 12 of infection these responses return to normal . In this study total and differential cell counts showed that as early as the third day of infection there was a marked reduction in the number of lymphocytes recovered from the peripheral blood, bone marrow, and thymus of infected animals . Concomitantly, there was an increase in the number of splenic lymphocytes . By day 28 both the total and differential cell counts were similar in both infected and normal animals . Flow microfluorometric (FMF) studies comparing the Thy-1.2, Lyt-1, Lyt-2, and surface immunoglobulin (slg) phenotypes of lymphocytes from normal and infected mice were performed . Between days 5 and 7 the thymocytes from infected mice displayed a higher relative fluorescence intensity (RFI) of the Thy-1.2 marker than normal thymocytes, whereas at day 10, the RFI was less than that of normal thymic lymphocytes . Between days 7 and 10 of infection the RFI of the Lyt-2 marker was less on thymocytes from Hc-infected mice; however, there was no change in the Lyt-1 marker . Examination of these lymphocyte markers in blood, spleen, and mesenteric lymph nodes showed that there were decreases in the RFI of both the Thy-1.2 and Lyt-2 between days 5 and 10 of infection . No changes were observed in the Lyt-1 or slg markers . By day 28 there were no differences between the normal and infected mice with respect to any surface marker in any of the organs studied . In other experiments, the effect of adrenalectomy before infection on these surface markers was studied . Absolute numbers of Thy-1.2+, Lyt-1+, and Lyt-2+ cells were significantly increased in the spleen and significantly decreased in the thymus and peripheral blood of infected mice relative to normal controls . These studies suggest that there is a migration of cells from the thymus, blood, and bone marrow to the spleens of mice with disseminated Hc infection.

J Biol Chem, 1983 Jul 25, 258(14), 8872 - 5
Reversal of the reaction catalyzed by glyoxalase I . Calculation of the equilibrium constant for the enzymatic reaction; Sellin S et al.; Glyoxalase I catalyzes the formation of S-D-lactoyl-glutathione via the hemimercaptal adduct of methylglyoxal and glutathione . This enzymatic reaction, which has been considered virtually irreversible, was found to be reversible under such conditions that glutathione liberated from the thiolester was trapped . The reverse reaction could be monitored spectrophotometrically by use of 5,5'-dithiobis-(2-nitrobenzoate) . In addition to 5,5'-dithiobis-(2-nitrobenzoate), 2,2'-dithiobispyridine and cystamine were used to promote the reverse reaction . S-D-Lactoylglutathione did not hydrolyze in the presence of glyoxalase I under the conditions investigated, as shown by its stability in the absence of thioltrapping agents . Proof of the reversal of the reaction was obtained by demonstrating the formation of stoichiometric amounts of methylglyoxal and glutathione from S-D-lactoylglutathione . Catalysis of the reverse reaction was dependent upon the presence of a bivalent metal ion in the active site of the enzyme . Apoenzyme, obtained by removal of the essential Zn2+ from the active site, did not catalyze the reverse reaction, but catalytic activity was restored by addition of Zn2+, Mg2+, Mn2+, or Co2+ . The reverse reaction was also catalyzed by glyoxalase I from yeast . Linear competitive inhibition (Ki = 0.64 mM) was obtained with 5,5'-dithiobis-(2-nitrobenzoate), which necessitated correction of the apparent kinetic parameters of the reverse reaction . The corrected values for the reverse reaction catalyzed by glyoxalase I from human erythrocytes with S-D-lactoylglutathione as substrate were kcat = 3.6 s-1 and Km = 1.9 mM . Combination of these values with the corresponding parameters for the forward reaction allowed calculation, through the Haldane relation, of the equilibrium constant, Keq = 1.1 X 10(4), for the isomerization between the hemimercaptal of methylglyoxal and glutathione and S-D-lactoylglutathione . The strong reversible competitive inhibitor of the forward reaction, S-p-bromobenzylglutathione, also inhibited the reverse reaction competitively (Ki = 0.38 microM).

J Biol Chem, 1983 Jul 10, 258(13), 8374 - 83
Enzymatic mechanism of an RNA ligase from wheat germ; Schwartz RC et al.; We have characterized the mechanism of action of a wheat germ RNA ligase which has been partially purified on the basis of its ability to participate in in vitro splicing of yeast tRNA precursors (Gegenheimer, P., Gabius, H-J., Peebles, C.L., and Abelson, J . (1983) J . Biol . Chem . 258, 8365-8373) . The preparation catalyzes the ligation of oligoribonucleotide substrates forming a 2'-phosphomonoester, 3',5'-phosphodiester linkage . The 5' terminus of an RNA substrate can have either a 5'-hydroxyl or a 5'-phosphate . The 5'-phosphate, which for a 5'-hydroxyl substrate can be introduced by a polynucleotide kinase activity in the preparation, is incorporated into the ligated junction . The 3' terminus can have either a 2',3'-cyclic phosphate or a 2'-phosphate . 2',3'-Cyclic phosphates can be converted into 2'-phosphates by a 2',3'-cyclic phosphate, 3'-phosphodiesterase activity in the preparation . The 2'-phosphate of the ligated product is derived from the phosphate at the 3' terminus of the substrate . Ligation proceeds with the adenylylation of the 5'-phosphorylated terminus to form an intermediate with a 5',5'-phosphoanhydride bond.

J Biol Chem, 1983 Jul 10, 258(13), 8365 - 73
An RNA ligase from wheat germ which participates in transfer RNA splicing in vitro; Gegenheimer P et al.; Transfer RNA half-molecules are intermediates in the splicing of tRNA precursors containing intervening sequences . We have utilized yeast tRNA half-molecules to identify and partially purify an ATP-dependent RNA ligase activity from extracts of wheat germ . This activity can complement a yeast tRNA endonuclease in vitro to efficiently splice 10 different yeast tRNA precursors . The products of in vitro splicing are a covalently joined tRNA and a circular intervening sequence RNA . The internucleotide bond formed at the splice junction is a 2'-phosphomonoester, 3',5'-phosphodiester structure . The 2'-phosphate originates from the 2',3'-cyclic phosphate at the 3' terminus of the 5' half-tRNA . The phosphodiester phosphate is derived from the gamma-phosphate of ATP.

J Biol Chem, 1983 Jul 10, 258(13), 8223 - 30
Sulfhydryl induced respiratory "shunt" pathways and their role in morphogenesis in the fungus, Histoplasma capsulatum; Sacco M et al.; When the mycelial to yeast transition of the dimorphic fungus Histoplasma capsulatum is induced by a temperature shift from 25 to 37 degrees C, the activities of the cytochrome system and the alternate oxidase decrease in parallel over the first 24 to 40 h (stage 1 of the transition) . The decrease in activity of the cytochrome system is correlated with extensive decreases in the amounts of cytochromes b, c, and aa3, assayed spectrophotometrically . After 40 h, the cells enter a dormant phase (stage 2 of the transition) and cysteine or other sulfhydryl-containing compounds are required to reactivate mitochondrial respiration . This reactivation is due to the establishment of shunt pathways which bypass blocked segments of the electron transport system . The "shunt" pathways operate normally in mycelia grown at 25 degrees C, but are shut down during the transition, possibly because of depletion of intracellular cysteine . The longstanding observation that cysteine is required to progress beyond the initial stages of the morphological transition may be due, at least in part, to the reactivation of these "shunt" pathways.

Biochemistry, 1983 Jul 5, 22(14), 3336 - 44
Phosphorus-31 nuclear magnetic resonance of ethidium complexes with ribonucleic acid model systems and phenylalanine-accepting transfer ribonucleic acid; Goldfield EM et al.; The temperature dependence of the 31P NMR spectra of the ethidium complexes with poly(A) X oligo(U) and the 31P spectra of phenylalanine tRNA (yeast) in various molar ratios of ethidium ion (Et) are presented . In the poly(A) X oligo(U) X Et complex, a new peak about 2.0 ppm downfield from the double-helix peak appears . We have assigned this peak to phosphates perturbed by ethidium . The chemical shift of this peak is consistent with the intercalation mode of binding and provides additional support for our hypothesis that 31P shifts are sensitive probes of phosphate ester conformations . The main effect of ethidium on the 31P spectra of tRNAPhe is the broadening of several of the scattered signals . These scattered signals are associated with phosphates involved in tertiary interactions . We propose that these broadened signals arise from phosphates near the Et binding site.

Am J Pathol, 1983 Jul, 112(1), 61 - 7
Correlation of regional disease and in vivo PO2 in rat mammary adenocarcinoma; Cole MA et al.; A knowledge of the distribution of oxygen tension (PO2) and vascularization in neoplasia has been fundamental to understanding relationships between tumor growth, hypoxia, and therapy . We have combined recessed oxygen microcathode and freeze-substitution techniques to correlate in situ PO2 profiles and morphologic features in 7,12-dimethylbenz(a)anthracene (DMBA) tumors in rats . Overlying connective tissue of transplanted tumor was exposed by a 1-2 mm incision and a cross-stitch pattern demarcated electrode puncture sites for histologic reference . Three buffered salt solutions (BSS) with different PO2 were each allowed to flow through a well over the tumor where electrodes were placed for calibration . Zero electrode oxygen current was recorded from a buffered yeast-agar mixture of zero torr . PO2 was recorded at 5-mu intervals to approximately 1-2 mm . Atmospheric contamination was eliminated by continuous well flow of BSS, 30 torr . Finally, the tumor and surrounding tissues were quick-frozen in vivo with Freon 22 and liquid nitrogen . The tissue block was freeze-substituted and sectioned . PO2 profiles were superimposed onto correspondingly scaled photomicrographs . A viable periphery with a PO2 range of 50-82 torr and a transition to necrotic areas of PO2, 2-13 torr were observed . This transition was characterized by PO2 gradients within distances of 50-300 mu at variable puncture depths . This technique should be useful in further studies of growth, necrosis, and therapy.

Prikl Biokhim Mikrobiol, 1983 Jul-Aug, 19(4), 498 - 502
{Effect of nitrogen sources on cellulase biosynthesis by a mutant strain of Trichoderma viride 44}; Ostrikova NA et al.; The effect of various nitrogen sources on cellulase biosynthesis by the mutant strain Trichoderma viride 44 was examined . This strain may utilized nitrogen in the nitrate, ammonium of organic form . When cultivating this strain, it appears advantageous to add to the nutrient medium yeast and yeast lyzates as well as their mixture with ammonium sulfate . Cellulase reached its maximum activity of 20.2, 21.5 and 23.2 mu/ml when grown on the medium containing ammonium phosphate, peptone and brewing yeast plus ammonium sulfate, respectively . It is useful to apply nitrogen in its organic forms in small quantities and in combination with mineral forms . The nitrogen presence in the medium is necessary only at the exponential stage of fungal growth . The lack of nitrogen in the stationary stage characterized by the maximum cellulase formation does not inhibit an increase in the enzyme activity.

J Biochem (Tokyo), 1983 Jul, 94(1), 71 - 7
Purification and properties of an endoribonuclease existing as a complex with inhibitor in rat liver cytosol; Kumagai H et al.; An endoribonuclease existing as a complex with inhibitor in the cytosol of rat liver has been purified about 128,000-fold after inactivation of the inhibitor with CdCl2 . The enzyme had a molecular weight of 16,000 and produced 3'-CMP via 2',3'-cyclic phosphate of cytidine from poly(C) . The breakdown of poly(U) by the enzyme was less than 5% of poly(C) breakdown . Poly(A) was not hydrolyzed by the enzyme . The enzyme had a pH optimum of 7.5-8, was heat-stable and had a Km of 952 micrograms yeast RNA and a Km of 198 micrograms poly(C) per ml . The maximal velocities for yeast RNA and poly(C) degradation were 3,970 A260/min/mg protein and 1,890 A260/min/mg protein, respectively . The enzyme was slightly stimulated by polyamines or monovalent and divalent cations except Mn2+, but was inhibited by nucleoside triphosphate, poly(G) and rat liver RNase inhibitor . Inhibition of the enzyme by rat liver RNase inhibitor was not prevented by monovalent and divalent cations or polyamines, although inhibition by poly(G) was prevented by these ions.

Infection, 1983 Jul-Aug, 11(4), 205 - 7
Bestatin treatment for the correction of granulocyte dysfunction in patients with recurrent furunculosis; Mattsson L et al.; Bestatin, a new immunomodulator which is chemically well-defined, was examined for its capacity to enhance the phagocytic activity of neutrophilic granulocytes from patients with furunculosis . The ability of the granulocytes to ingest fluorescein-labelled yeast particles was significantly decreased in 19 patients with recurrent furunculosis (p less than 0.01) . Oral administration of 40 mg bestatin to ten patients increased the phagocytic function of their granulocytes significantly (p less than 0.01).

Nippon Yakurigaku Zasshi, 1983 Jul, 82(1), 27 - 35
{Effect of cysteine ethylester hydrochloride (Cystanin) on host defense mechanism}; Goto K et al.; Cysteine ethylester (10 approximately 100 mg/kg, p.o.) augmented the production of hemolytic plaque-forming cells (HPFC) to sheep red blood cells and lipopolysaccharide in the spleen of mice . It showed no effect, however, on the HPFC production to trinitrophenylated polyvinylpyrrolidone; and it had no activity as a polyclonal B cell activator like lipopolysaccharide . Cysteine ethylester at a concentration of 3 microM or more potentiated the phagocytosis of yeast by the peritoneal polymorphonuclear leukocytes of rats . This activity was less potent than that of levamisole and D-penicillamine . It also potentiated the reduction of nitroblue tetrazolium (NBT) by blood leukocytes prepared from guinea pigs and a human being . This activity was more potent than that of levamisole and D-penicillamine . Furthermore, cysteine ethylester at doses showing an immunopotentiating activity protected irradiated mice from death by spontaneous infection . These findings suggest that this drug may have a capacity to potentiate the host defense mechanisms.

Nature, 1983 Jun 30, 303(5920), 806 - 8
Possible relationship of morphogenesis in pathogenic fungus, Histoplasma capsulatum, to heat shock response; Lambowitz AM et al.; Histoplasma capsulatum, like many other fungal pathogens, is dimorphic: it exists as mycelia in the soil and yeast in animal hosts . Because only the yeast phase is parasitic, factors which affect morphogenesis have been of interest for understanding and controlling pathogenicity . In culture, the mycelial to yeast transition of H . capsulatum is induced by a temperature shift from 25 to 37 degrees C (ref . 1) . The transition occurs over several days and is accompanied by marked changes in metabolic processes, including respiration and cysteine metabolism . Here, we show that the triggering event for these morphological and biochemical changes is a rapid decline in intracellular ATP levels that follows uncoupling of oxidative phosphorylation when mycelia are shifted from 25 to 37 degrees C . We also show that respiration in the yeast phase is coupled at 37 degrees C and thus that the morphological transition may be viewed as a heat shock followed by cellular adaptation to higher temperature.

J Biol Chem, 1983 Jun 25, 258(12), 7702 - 6
Studies on the effect of ethanol on eukaryotic protein synthesis in vitro; David ET et al.; The effects of varying concentrations of ethanol on reactions involved in protein biosynthesis have been examined using a cell-free system from Chinese hamster ovary cells that actively translates natural mRNAs in order to detect those components most sensitive to alcohol . Ethanol, at relatively low concentrations (0.2 M or lower) inhibited the translation of endogenous polysomal mRNAs and, in mRNA-depleted extracts, of exogenous natural mRNA . Ethanol markedly inhibited leucyl-tRNA synthetase, and it inhibited Phe- and Glu-tRNA synthetases to some extent, but had only a small effect on several other aminoacyl-tRNA synthetases, elongation factors 1 and 2, ribosomes, or the formation of eukaryotic initiation factor 2 . GTP . Met-tRNAr ternary complex . Methanol inhibited slightly the translation of mRNA and Leu-tRNA synthetase, but isobutyl alcohol and isopropyl alcohol strongly depressed these activities . Ethanol inhibited the interaction of leucine with Leu-tRNA synthetase competitively, whereas isobutyl alcohol and acetaldehyde inhibited the leucine interaction in a noncompetitive manner . Leu-tRNA synthetase from Chinese hamster ovary cells was more sensitive to ethanol than that from yeast.

Nucleic Acids Res, 1983 Jun 25, 11(12), 4211 - 27
Nucleotide sequence of a segment of Drosophila mitochondrial DNA that contains the genes for cytochrome c oxidase subunits II and III and ATPase subunit 6; Clary DO et al.; The nucleotide sequence of a segment of the mtDNA molecule of Drosophila yakuba has been determined, within which have been identified the genes for tRNAleuUUR, cytochrome c oxidase subunit II (COII), tRNAlys, tRNAasp, URFA6L, ATPase subunit 6 (ATPase6), cytochrome c oxidase subunit III (COIII) and tRNAgly . The genes are arranged in the order given and all are transcribed from the same strand of the molecule in a direction opposite to that in which replication proceeds around the molecule . The tRNAlys gene is unusual among mitochondrial tRNAlys genes in that it contains a CTT anticodon . The triplet AGA is used to specify an amino acid in all of the COII, COIII, ATPase6, and URFA6L genes . However, the AGA codons found in these four polypeptide genes correspond in position to codons which specify nine different amino acids, but never arginine, in the equivalent polypeptide gene which have been sequenced from mtDNAs of mouse, yeast and Zea mays.

Nucleic Acids Res, 1983 Jun 25, 11(12), 3903 - 17
Multiple, independently regulated, polyadenylated messages for histone H3 and H4 in Tetrahymena; Bannon GA et al.; Heterologous probes for yeast H4 and H3 histone genes have been used to study the corresponding histone mRNAs in growing and starved Tetrahymena . Histone mRNAs in both physiological states are polyadenylated . Two types of H4 protein and two types of H3 protein have previously identified in Tetrahymena . Two size classes of H4 messages and three classes of H3 messages have been detected by northern analyses . Southern blot analysis indicate that the number of different kinds of H3 and H4 genes is the same or slightly greater than the number of different messages, suggesting that each message is derived from a different gene . Growing cells have -30 times more histone mRNA than starved cells, even though their total mRNA content is only 4 times greater . The relative abundance of different H4 and H3 messages in growing and starved cells is different, demonstrating that the different messages for a particular type of histone are regulated non-coordinately . In starved cells the presence of a single size class of H3 messages correlates with the preferential synthesis of a previously described macronuclear-specific H3 variant . The fraction of histone messages loaded in growing and starved cells is the same as for bulk mRNAs, and the relative concentrations of the multiple messages for H4 and H3 are the same in polysomal and total RNAs of each cell type . These observations suggest that histone synthesis in Tetrahymena is controlled largely at the level of message abundance, and that very little, if any, control occurs at the translational level.

Biochemistry, 1983 Jun 21, 22(13), 3178 - 87
Lipid and subunit III depleted cytochrome c oxidase purified by horse cytochrome c affinity chromatography in lauryl maltoside; Thompson DA et al.; Cytochrome oxidase is purified from rat liver and beef heart by affinity chromatography on a matrix of horse cytochrome c-Sepharose 4B . The success of this procedure, which employs a matrix previously found ineffective with beef or yeast oxidase, is attributed to thorough dispersion of the enzyme with nonionic detergent and a low density of cross-linking between the lysine residues of cytochrome c and the cyanogen bromide activated Sepharose . Beef heart oxidase is purified in one step from mitochondrial membranes solubilized with lauryl maltoside, yielding an enzyme of purity comparable to that obtained on a yeast cytochrome c matrix {Azzi, A., Bill, K., & Broger, C . (1982) Proc . Natl . Acad . Sci . U.S.A . 79, 2447-2450} . Rat liver oxidase is prepared by hydroxyapatite and horse cytochrome c affinity chromatography in lauryl maltoside, yielding enzyme of high purity (12.5-13.5 nmol of heme a/mg of protein), high activity (TN = 270-400 s-1), and very low lipid content (1 mol of DPG and 1 mol of PI per mol of aa3) . The activity of the enzyme is characterized by two kinetic phases, and electron transfer can be stimulated to maximal rates as high as 650 s-1 when supplemented with asolectin vesicles . The rat liver oxidase purified by this method does not contain the polypeptide designated as subunit III . Comparisons of the kinetic behavior of the enzyme in intact membranes, solubilized membranes, and the purified delipidated form reveal complex changes in kinetic parameters accompanying the changes in state and assay conditions, but do not support previous suggestions that subunit III is a critical factor in the binding of cytochrome c at the high-affinity site on oxidase or that cardiolipin is essential for the low-affinity interaction of cytochrome c . The purified rat liver oxidase retains the ability to exhibit respiratory control when reconstituted into phospholipid vesicles, providing definitive evidence that subunit III is not solely responsible for the ability of cytochrome oxidase to produce or respond to a membrane potential or proton gradient.

Cryobiology, 1983 Jun, 20(3), 298 - 309
Ice nucleation and freezing in undercooled cells; Franks F et al.; DSC has been employed to study the effect of cooling on a range of cells under exclusion of extracellular ice and in the absence of chemical cryoprotectants . In contrast to earlier reports, all the cells studied were found to freeze at temperatures above that indicated for homogeneous nucleation of ice in undercooled liquid water . In the case of human erythrocytes this temperature difference was only 0.5 degrees, but for yeast cells and cells of plant origin the difference amounted to congruent to 9 degrees . Nucleation of ice within the cell (or at the cell wall/membrane) must therefore be initiated by a heterogeneous mechanism . A kinetic analysis of the temperature dependence of nucleation shows the rates to be consistent with the dimensions of the plant cells (or organelles), if these were to be the active nucleators . However, the nucleation kinetics of human erythrocytes are extremely temperature sensitive, and the kinetic parameters only differ by small, though significant, extents from those of the suspension medium . Possible nucleation mechanisms are discussed in terms of the experimental data and the cell dimensions . Finally, one of the underlying assumptions of the kinetic analysis, i.e., that ice growth must be rapid compared to nucleation, has been tested and validated by freeze-fracture electron microscopy.

Sabouraudia, 1983 Jun, 21(2), 121 - 7
Study of equine histoplasmosis farciminosi and characterization of Histoplasma farciminosum; Gabal MA et al.; A detailed clinical and mycological study of horse infections with Histoplasma farciminosum was conducted for the first time in the Middle East . The disease seems to prevail in endemic form in the region . In all of the cases studied the infection involved only the cutaneous lymphatics and skin tissue with extension to the regional draining lymph glands . The disease seems to impose serious economic impact in the infected areas . Full description and thorough characterization of both the mycelial form and the yeast phase of the causative fungus were made.

J Pharm Sci, 1983 Jun, 72(6), 626 - 30
Antitumor agents LIX: effects of quassinoids on protein synthesis of a number of murine tumors and normal cells; Hall IH et al.; The quassinoids (brusatol, bruceantin, bisbrusatolyl esters, and bisbruceantinyl esters of succinic and malonic acids) were observed not to be universal protein synthesis inhibitors . Rather, they were selective for both the types of cancers, e.g., P-388 lymphocytic leukemia, Ehrlich and hepatoma carcinoma and L-1210 lymphoid leukemia, as well as types of normal tissues (e.g., lymphocytes), in which they demonstrated protein synthesis inhibition . The data suggest that the observed difference in the magnitude of protein synthesis inhibition of two P-388 lymphocytic leukemia cell lines by the quassinoids was at the ribosomal levels, whereas the observed difference in normal livers from various strains of mice involve differences in cell membrane transport of the quassinoids into the various tissues . Detailed studies indicated that the mode of action of the quassinoids as protein synthesis inhibitors was identical in all of the cells where inhibition was observed; i.e., the elongation step of protein synthesis was blocked by the quassinoids . The data derived from assays for polyuridine-directed polyphenylalanine synthesis of isolated ribosomes demonstrated that the ID50 values obtained were consistent with the observed inhibition of whole cell protein synthesis inhibition for P-388 cells, as well as for BDF1 and DBA/2 liver cells . The ID50 values obtained from various cells, e.g., P-388 cells and normal liver, were all in the microM range and are consistent with previously published values for the quassinoids in the rabbit reticulocyte and yeast systems.

Am J Clin Nutr, 1983 Jun, 37(6), 887 - 97
Bioavailability of selenium to Finnish men as assessed by platelet glutathione peroxidase activity and other blood parameters; Levander OA et al.; Three groups of 10 men of low selenium status were given 200 micrograms Se/day as Serich wheat, Se-rich yeast, or sodium selenate for 11 wk . Twenty unsupplemented subjects served as controls . Plasma Se levels increased steadily in the wheat and yeast groups for 11 wk without plateauing, whereas in the selenate group, plasma Se plateaued around 110 ng/ml after 4 wk . Platelet glutathione peroxidase (GSH-Px) activities increased rapidly in the wheat and selenate groups for 4 wk and then plateaued . Platelet GSH-Px increased more slowly in the yeast group . Ten weeks after the supplements were discontinued, platelet GSH-Px was higher in the wheat and yeast groups than in the selenate group . Assessment of Se bioavailability requires a short-term platelet GSH-Px measurement to determine immediate availability, a medium-term plasma Se measurement to estimate retention, and a long-term platelet GSH-Px measurement after supplements are discontinued to determine the covertibility of tissue Se stores to biologically active Se.

Nippon Yakurigaku Zasshi, 1983 Jun, 81(6), 481 - 92
{Pharmacological studies of dl-2{3-(2'-chlorophenoxy)phenyl}propionic acid . I . Analgesic and antipyretic effects}; Kaneko T et al.; Analgesic and antipyretic effects of dl-2{3-(2'-chlorophenoxy)phenyl} propionic acid (CPP) were studied in mice, rats and guinea-pigs . CPP produced a dose dependent inhibition of acetic acid-induced writhing syndrome . Its ED50 values 1 and 3 hr after oral administration were 47 and 31 mg/kg, respectively . CPP had a potent analgesic effect on bradykinin-induced nociceptive response in rats, and its ED50 value was 15 mg/kg 2 hr after oral administration . The analgesic activity of CPP in these experiments was less potent than that of indomethacin, but it was approximately equivalent to ibuprofen and 10 to 20 times as potent as aspirin . CPP had no analgesic effect on both the tail pinch and hot plate tests in mice, while CPP potentiated the analgesic effect of codeine on these tests . CPP had no effect on the nociceptive response induced by intradermal injection of bradykinin and/or EDTA in guinea-pigs . On the other hand, when CPP was given orally in a dose range of 1.25 to 5 mg/kg, it produced an antipyretic effect on yeast-induced fever in rats . The antipyretic activity of CPP was equivalent to ibuprofen and 10 to 15 times as potent as aspirin.

Ann Intern Med, 1983 Jun, 98(6), 958 - 72
Genital herpes simplex virus infections: clinical manifestations, course, and complications; Corey L et al.; The clinical course and complications of 268 patients with first episodes and 362 with recurrent episodes of genital herpes infection were reviewed . Symptoms of genital herpes were more severe in women than in men . Primary first-episode genital herpes was accompanied by systemic symptoms (67%), local pain and itching (98%), dysuria (63%), and tender adenopathy (80%) . Patients presented with several bilaterally distributed postular ulcerative lesions that lasted a mean of 19.0 days . Herpes simplex virus was isolated from the urethra, cervix, and pharynx of 82%, 88%, and 13% of women with first-episode primary genital herpes, and the urethra and pharynx of 28% and 7% of men . Complications included aseptic meningitis (8%), sacral autonomic nervous system dysfunction (2%), development of extragenital lesions (20%), and secondary yeast infections (11%) . Recurrent episodes were characterized by small vesicular or ulcerative unilaterally distributed lesions that lasted a mean of 10.1 days . Systemic symptoms were uncommon and 25% of recurrent episodes were asymptomatic . The major concerns of patients were the frequency of recurrences and fear of transmitting infection to partners or infants.

Immunol Lett, 1983 Jun, 6(6), 287 - 91
C3b acceptors on macrophages: inhibition of Fc gamma-receptor-mediated phagocytosis by acceptor-bound C3b; Fabry Z et al.; The binding of nascent human C3b (i.e . the fragment of C3 just after trypsin cleavage) to mouse peritoneal macrophages was demonstrated by immune adherence . Acceptor-bound C3b could be detected longer than 24 h on the cell membrane . The rosette formation and phagocytosis of SRBC coated with anti-SRBC rat IgG was inhibited by preincubation of the cells with C3 and trypsin (15 min, 37 degrees C) . However, the phagocytosis of opsonized yeast particles was not influenced by acceptor-bound C3b, proving that C3b-C3b acceptor interaction did not alter the function of C3b-receptors . Acceptor-bound C3b on the macrophages failed to mediate phagocytosis of human 0,Rh+ red cells having C3b-receptors.

Cell Biophys, 1983 Jun, 5(2), 119 - 28
Lateral diffusion of wheat germ agglutinin-labeled glycoconjugates in the membrane of differentiating HL-60 and U-937 cells assessed with fluorescence recovery after photobleaching (FRAP); Magnusson KE et al.; The promyelocytic leukemia cell line HL-60 and the histiocytic cell line U-937 were grown in suspension culture . They were induced to differentiate during 5-d cultivation in the presence of dimethylsulfoxide (DMSO; 1.3% w/v) or phorbol-12-myristate-acetate (PMA; 10(-7) M), which yields granulocyte- and macrophage-like cells, respectively . Differentiation was evidenced by increased capacity to recognize and phagocytize IgG- or complement-coated yeast particles . Aliquots taken from the cultures with and without DMSO (or PMA) were spun down directly on glass microscope slides, washed, labeled with fluoresceinated wheat germ agglutinin (WGA), and directly examined at room temperature for the rate of fluorescence recovery after photobleaching (FRAP) . It was found that cultivation of the HL-60 and the U-937 cells in the presence of DMSO, which yields granulocyte-like cells, reduced the average value of lateral diffusion coefficient D (X 10(10} from 1.72 +/- 0.13 cm2s-1 to 0.97 +/- 0.13 cm2s-1 and from 1.77 +/- 0.11 cm2s-1 to 0.82 +/- 0.13 cm2s-1, respectively . U-937 cells grown with PMA also showed a reduction of D(X 10(10} to 0.88 +/- 0.10 cm2s-1 . There was a larger immobile fraction of fluorescence in the HL-60 cells than in the U-937 cells, viz., 70-80% compared to 10-50% . The total number of binding sites for WGA was not altered, but the surface density changed, since the HL-60 and the U-937 cells became smaller and larger, respectively, when grown in the presence of DMSO . It is concluded that differentiation reduces the average lateral mobility of the WGA-binding membrane component by a factor around 2.

J Immunol Methods, 1983 May 27, 60(1-2), 115 - 24
Differentiation between attached and ingested immune complexes by a fluorescence quenching cytofluorometric assay; Sahlin S et al.; Immune complexes attached to and ingested by human polymorphonuclear (PMN) cells were quantified by cytofluorometry using a fluorescence quenching assay which permits differentiation between attachment and ingestion . The fluorescence intensity decreased after ingestion as a result of the low pH in the phagolysosomes . When extracellular pH was lowered a slight decrease in phagolysosomal pH was detected in macrophages but not in PMN . When measuring total fluorescence, interaction at pH 5.8 for PMN and at pH 4.4 for macrophages is recommended, since the intensity of extra- and intracellular fluorescence are equal under these conditions . Thirty different dyes were tested for dye exclusion and fluorescence quenching of FITC-conjugated yeast particles, and FITC-conjugated IgG . Because of the lysosomotropic effect of basic dyes, acid and direct dyes are preferable as quenching agents . We could not find physical or chemical properties of the dyes that correlated with their quenching effect . Heat aggregated IgG was used as an immune complex analogue in the development of the assay . Trypan blue (0.2 mg/ml) at pH 4.4 was found to be the best quenching agent of extracellular fluorescence when using ingested aggregated IgG . The technique offers a simple method of quantifying ingested protein aggregates and of studying heterogeneity in phagocyte populations.

Eur J Biochem, 1983 May 16, 132(3), 525 - 30
Phosphorylated intermediate of a transport ATPase and activity of protein kinase in membranes from corn roots; Scalla R et al.; A maize-root microsomal fraction was enriched in ATPase by treatment with Triton X-100 . This activity, which reached 1.2-2.0/mumol Pi x min-1 x mg protein-1, was specific for ATP, very slightly stimulated by K+, inhibited by orthovanadate and diethylstilbestrol, resistant to oligomycin and azide, and had a Km of 1.2 mM MgATP . Incubation of the microsomal fraction with {gamma 32-P}ATP followed by electrophoresis in acid conditions revealed the presence of several phosphoproteins . The phosphorylation of a 110000-Mr polypeptide reached the steady-state level in less than 5 s and rapidly turned over the phosphate group . The phosphorylation level was an hyperbolic function of the {ATP} with a Km of 0.6 mM, suggesting that the rate of Pi production was proportional to the phosphoprotein concentration . The extent of phosphoprotein was decreased by vanadate and diethylstilbestrol . The phosphorylation level was 30% decreased by 50 mM K+ or Na+ while the ATPase activity was slightly stimulated (12% and 5%, respectively) . The polypeptide could not be phosphorylated in reverse by Pi . This phosphorylated intermediate from maize-root microsomes exhibits molecular properties characteristic of transport ATPases such as the yeast plasma membrane H+-translocating ATPase . This similarity indicates existence of a transport ATPase in plant plasma membranes . Three other plant microsomal polypeptides (Mr = 52000, 17000 and 16000) and a low molecular weight component (Mr less than 1000) were phosphorylated much more slowly, were not undergoing a rapid turnover and were not hydrolysed by hydroxylamine . These phosphoproteins and the Mr less than 1000 phosphorylated component were inhibited by vanadate and diethylstilbestrol . These properties are similar to those of the protein kinase activity recently described in yeast plasma membranes.

Nucleic Acids Res, 1983 May 11, 11(9), 2765 - 78
Two Ustilago maydis viral dsRNAs of different size code for the same product; Field LJ et al.; UmV is a double-stranded RNA (dsRNA) virus of the corn fungus Ustilago maydis . There are three viral subtypes, P1, P4 and P6, which differ in the specificity of their secreted killer toxins . Each has three size classes of dsRNAs: H (heavy), M (medium), and L (light) . We find that, unique among dsRNA viruses, two segments of different size code for the same product--the toxin resistance factor . The smaller dsRNA (L) is homologous to one end of the larger (M), and may have arisen by replication and packaging of a sub-genomic mRNA . We have also compared all the UmV dsRNAs with each other and with the dsRNAs of the similar yeast virus (ScV) by Northern gel and by 3' sequence analysis . Like those of ScV, many of the UmV dsRNAs have one 3' terminus with the sequence UUUUUCAOH or UUUUUCGOH . The H and L dsRNAs of similar size in different viral subtypes are generally related in sequence . The UmV H dsRNAs of different size are not detectably related in sequence.

J Infect Dis, 1983 May, 147(5), 824 - 8
Three-step stool examination for cryptosporidiosis in 10 homosexual men with protracted watery diarrhea; Ma P et al.; Cryptosporidiosis, a zoonosis caused by Cryptosporidium species, is a newly recognized coccidial protozoan infection causing severe protracted watery diarrhea in humans . In August 1981, the first case of cryptosporidiosis in a homosexual man with acquired immune deficiency syndrome (AIDS) was reported; diagnosis was determined by intestinal biopsy . It is necessary to adopt a simple laboratory diagnostic procedure to screen large numbers of suspected cases . A three-step stool examination was developed to demonstrate Cryptosporidium oocysts and the diagnostic and infective stages of the infection in 10 homosexual men with AIDS . This is a less invasive, less costly, and more sensitive test than intestinal biopsy and has been designed to prevent confusion caused by yeast cells that are frequently present in stool, leading to a false diagnosis . The examination consists of preliminary differential determination by iodine wet mount, definitive identification by modified Kinyoun acid-fast staining, and a more effective method of concentrating oocysts, by Sheather's sugar cover-slip flotation method.

Allergol Immunopathol (Madr), 1983 May-Jun, 11(3), 195 - 200
Annual fluctuations of mites and fungi in Danish house-dust: an example; Hallas TE et al.; For one year dust was sampled monthly from a single mattress . Until August mite counts were at background levels . In September and October there was a burst in the counts for the house-dust mite, Dermatophagoides farinae and for the house mite, Glycyphagus domesticus . Their occurrence might be interpreted as a response to large densities of an unidentified yeast in September . This yeast appeared when the indoor absolute humidity exceeded 9 g H2O/kg dry air . The mite Tarsonemus rakowiensis reached its maximum in October, and might be a possible house-dust mite predator, although the evidence is far from conclusive . An aliquot of ten grams of mattress dust sampled from a mite-free location on top of the mattress hardly became infested, although the environmental conditions were ideal . Obviously, the mattress dust from the dry room was not optimal for either the yeast or the mites.

J Clin Microbiol, 1983 May, 17(5), 909 - 17
Factors influencing the reactivity of Legionella antigens in immunofluorescence tests; Benson RF et al.; We examined several factors for their effects on the serological reactivity of Legionella antigens used for direct or indirect fluorescent-antibody tests . These factors included media, methods of killing, strain differences, and the nature of the reactivity with diverse human sera . The maximum serological reactivities were obtained with charcoal-yeast extract agar; the relative antigenicity of cells grown on a chemically defined medium could be fourfold less than those grown on the charcoal-yeast extract agar . Cells grown at 25 degrees C showed only small antigenic differences from those grown at 35 degrees C but had better morphological and staining characteristics . Cells killed by 1% Formalin or 37% Formalin vapors showed a 20% less relative antigenicity than those killed by heat, but their cell walls stained more clearly and they had fewer aberrations . As tested with several human sera, cells of Philadelphia 1 showed great variation in relative antigenicity with changes in media or methods of preparation; Bellingham 1 was quite stable under these same conditions . The data suggest that Bellingham 1 had serogroup 1-specific antigens, reactive with human sera, which were not present in Philadelphia 1.

Infect Immun, 1983 Apr, 40(1), 417 - 20
Susceptibility of congenitally athymic (nude) mice to sporotrichosis; Dickerson CL et al.; Congenitally athymic (nu/nu) mice were found to be more susceptible to intravenous challenge with Sporothrix schenckii than their phenotypically normal (nu/+) littermates as measured by lethality and the number of viable yeast cells in the liver 7 days postinfection . Thymus reconstitution of nu/nu mice (nu/thy) conferred a significant degree of resistance to sporotrichosis . Immunization greatly enhanced the resistance of nu/thy and nu/+ mice, but unexpectedly increased the susceptibility of nu/nu mice . The susceptibility of nonimmunized nu/nu mice and the finding that thymus transplants augmented resistance to sporotrichosis suggest that T lymphocytes are critical to host defense.

Proc Natl Acad Sci U S A, 1983 Apr, 80(7), 1811 - 5
In vitro synthesis of thymosin beta 4 encoded by rat spleen mRNA; Filipowicz AW et al.; Thymosin beta 4, containing 43 amino acids and acetylated at the NH2 terminus, is synthesized in vitro in a rabbit reticulocyte lysate or in a yeast protein-synthesis system in the presence of mRNA from rat spleen . The product formed was identified as beta 4 by immunoprecipitation by a specific anti-beta 4 antiserum, comigration with authentic beta 4 in NaDodSO4/polyacrylamide gel electrophoresis and in HPLC, and identity of peptide fragments . The immunoprecipitable product generated in the wheat germ protein-synthesizing system emerged slightly ahead of beta 4 in HPLC and appeared to lack the NH2-terminal acetyl group . There was no evidence for formation of a larger polypeptide precursor of beta 4 in any of the three systems used . In sucrose density gradient centrifugation, the mRNA coding for beta 4 was recovered in the 7-8S mRNA fraction.

Zentralbl Bakteriol Mikrobiol Hyg {B}, 1983 Apr, 177(3-4), 327 - 33
Viral pollution of some artificial reservoirs in Romania; Nestor I et al.; Five reservoirs were investigated for the presence of enteroviruses by using simultaneously two methods for concentration (PE60 and yeast cells) and two techniques for isolation (inoculation into suckling mice and into cell cultures) . The average rate of virus positive samples was lower in reservoir water than in the water of the adjoined rivers . The paper discusses the possibilities of viral self-purification in reservoir water and the advantages of its use for drinking purposes after adequate treatment.

Nucleic Acids Res, 1983 Mar 25, 11(6), 1759 - 71
The nucleotide sequence of the rat cytoplasmic beta-actin gene; Nudel U et al.; The nucleotide sequence of the rat beta-actin gene was determined . The gene codes for a protein identical to the bovine beta-actin . It has a large intron in the 5' untranslated region 6 nucleotides upstream from the initiator ATG, and 4 introns in the coding region at codons specifying amino acids 41/42, 121/122, 267, and 327/328 . Unlike the skeletal muscle actin gene and many other actin genes, the beta-actin gene lacks the codon for Cys between the initiator ATG and the codon for the N-terminal amino acid of the mature protein . The usage of synonymous codons in the beta-actin gene is nonrandom, and is similar to that in the rat skeletal muscle and other vertebrate actin genes, but differs from the codon usage in yeast and soybean actin genes.

J Biol Chem, 1983 Mar 25, 258(6), 3865 - 72
Pyrroline-5-carboxylate synthesis from glutamate by rat intestinal mucosa; Wakabayashi Y et al.; The mitochondria of rat intestinal mucosa were found to have an enzymatic activity that converts radioactive glutamate to pyrroline-5-carboxylate (P5C) in the presence of ATP, NADPH, and MgCl2 . The product of this enzyme was identified as P5C by the fact that it was converted to proline by chemical reduction with NaBH4 or by enzymatic reduction with NADH in the presence of purified yeast P5C reductase . The product was demonstrated to be P5C rather than pyrroline-2-carboxylate by thin layer chromatography . The presence of the activity in mitochondria prepared from intestinal mucosa of germ-free rats proved that this activity is of mammalian origin . Omission of either ATP, NADPH, or MgCl2 from the reaction mixture resulted in little or no activity . The optimal pH appeared to be about 7.0 under the conditions used . Substrate saturation curves in the presence of an ATP and an NADPH regeneration system gave apparent Km values of 2.5 mM for glutamate, 0.19 mM for ATP, and 6.5 microM for NADPH in the presence of 20 mM MgCl2 . The mitochondrial preparation usually produced P5C at a rate of 1.2 to 1.6 nmol/mg/min at 20 degrees C when incubated with 1 mM glutamate, 3 mM ATP, 0.2 mM NADPH, and 20 mM MgCl2.

Eur J Biochem, 1983 Mar 15, 131(2), 349 - 52
Interaction between isolated cytochrome c1 and cytochrome c; Broger C et al.; Cytochrome c1 from bovine heart mitochondria was isolated by a modification of the technique of Konig et al . {(1980) Biochim . Biophys . Acta 621, 283-295} which involved an affinity chromatography step on a gel with yeast cytochrome c as a ligand . Its spectra, electrophoretic pattern in presence of sodium dodecylsulfate, its reducibility by ascorbate and cytochrome c were characteristic of a native cytochrome, with a single polypeptide having an apparent molecular weight of 30 000 . By using an arylazido derivative of cytochrome c, having the photoactive group bound to lysine 13, upon illumination a cross-link with the described preparation of cytochrome c1 was obtained . By pepsin digestion of the cross-linked complex a limiting fragment was obtained and partially sequenced . It allowed to identify the site of binding of cytochrome c near the sequence 167-174 of cytochrome c1.

J Infect Dis, 1983 Mar, 147(3), 504 - 13
Serum arabinitol concentrations and arabinitol/creatinine ratios in invasive candidiasis; Gold JW et al.; Serum arabinitol concentrations and arabinitol/creatinine ratios were determined in 25 cancer patients with invasive candidiasis, 73 severely ill cancer patients who did not have invasive candidiasis, and 15 uninfected patients with chronic renal failure . Elevated arabinitol concentrations were found in patients with renal failure and invasive candidiasis . Serum arabinitol concentrations exceeded 1.19 micrograms/ml in 13 of 18 patients with invasive candidiasis who were studied when renal function was normal and in three of 52 control patients . Among patients with renal failure, 10 of 14 with invasive candidiasis but only four of 36 control patients had serum arabinitol concentrations of greater than 5.85 micrograms/ml . Serum arabinitol and creatinine concentrations were strongly correlated . The arabinitol/ creatinine ratio exceeded 1.5 in 16 of 25 patients with invasive candidiasis but in only three of 88 control patients . Increased serum arabinitol concentrations appear to reflect increased production of arabinitol by yeast in individuals with invasive candidiasis.

J Am Acad Dermatol, 1983 Mar, 8(3), 378 - 85
Decreased release of lysosomal enzymes from peripheral leukocytes of patients with atopic dermatitis; Ring J et al.; Peripheral blood leukocytes from fifteen patients with atopic dermatitis and ten normal nonatopic volunteers were incubated with various stimuli in vitro, and the release of the lysosomal beta-glucuronidase into the supernatant was measured . beta-Glucuronidase release was significantly reduced in patients with severe atopic dermatitis after stimulation with aggregated IgG, horse antihuman lymphocyte globulin (ALG), zymosan, and yeast-activated serum . There was an indirect correlation (r = -0.83) between aggregated IgG-induced beta-glucuronidase release and the intensity of clinical symptoms; however, there was no correlation with serum IgE levels . The enzyme release measured was not caused by cellular lysis, except for high concentrations of antilymphocyte globulin, as determined by lactic dehydrogenase (LDH) activity in the supernatant . It is concluded that lysosomal enzyme release defects might be involved in the well-known decreased resistance to infections in patients with atopic dermatitis.

Proc Natl Acad Sci U S A, 1983 Mar, 80(5), 1241 - 5
Rates of enzyme-catalyzed exchange determined by two-dimensional NMR: a study of glucose 6-phosphate anomerization and isomerization; Balaban RS et al.; The application of two-dimensional (2D) Fourier-transform NMR to the determination of rate constants of complex enzyme-catalyzed reactions in the steady state is described . The yeast phosphoglucose isomerase (EC 5.3.1.9)-catalyzed anomerization of glucose 6-phosphate (Glc-6-P) as well as its isomerization to fructose 6-phosphate (Fru-6-P) was chosen as an example . The 2D technique permitted the simultaneous monitoring of the time course of the anomerization and isomerization steps, from which the various reaction rates were determined . The results obtained in the steady state demonstrate the usefulness of the 2D technique by confirming that the anomerization of Glc-6-P is enzyme catalyzed and that the isomerization of the alpha anomer of Glc-6-P to Fru-6-P is at least 10 times faster than the isomerization of the beta anomer of Glc-6-P . These results are compared with reaction rates obtained by rapid-quench methods and the mechanistic implications are discussed . Extrapolation of these results suggests that the 2D Fourier-transform NMR method should be applicable in intact biological tissues.

Arch Biochem Biophys, 1983 Mar, 221(2), 593 - 7
Castanospermine, a tetrahydroxylated alkaloid that inhibits beta-glucosidase and beta-glucocerebrosidase; Saul R et al.; Castanospermine (1,6,7,8-tetrahydroxyoctahydroindolizine) was tested against a variety of commercially available glycosidases and found to be a potent inhibitor of almond emulsin beta-glucosidase, and also to inhibit fungal beta-xylosidase . This alkaloid was inactive on yeast alpha-glucosidase, alpha- or beta-galactosidase, alpha-mannosidase, beta-N-acetylhexosaminidase, beta-glucuronidase, alpha-L-fucosidase . Fifty-percent inhibition of beta-glucosidase required about 10 micrograms/ml of castanospermine . The amount of inhibition was uniform throughout the time course, and the inhibition with regard to substrate concentration (p-nitrophenyl-beta-D-glucopyranoside) appeared to be of the mixed type . Castanospermine was also a potent inhibitor of beta-glucocerebrosidase when assayed with fibroblast extracts using either a fluorimetric or a radioactive assay . Interestingly enough, castanospermine also inhibited the lysosomal alpha-glucosidase, and this inhibition required comparable levels of alkaloid to that required for inhibition of beta-glucocerebrosidase . However, a number of other lysosomal glycosidases were not sensitive to castanospermine (i.e., alpha- or beta-galactosidase, alpha- or beta-mannosidase, alpha- or beta-L-fucosidase, beta-N-acetylhexosaminidase, beta-glucuronidase).

Appl Environ Microbiol, 1983 Mar, 45(3), 784 - 91
Production of superoxide and hydrogen peroxide in medium used to culture Legionella pneumophila: catalytic decomposition by charcoal; Hoffman PS et al.; The difficulties associated with the growth of Legionella species in common laboratory media may be due to the sensitivity of these organisms to low levels of hydrogen peroxide and superoxide radicals . Exposure of yeast extract (YE) broth to fluorescent light generated superoxide radicals (3 microM/h) and hydrogen peroxide (16 microM/h) . Autoclaved YE medium was more prone to photochemical oxidation than YE medium sterilized by filtration . Activated charcoals and, to a lesser extent, graphite, but not starch, prevented photochemical oxidation of YE medium, decomposed hydrogen peroxide and superoxide radicals, and prevented light-accelerated autooxidation of cysteine . Also, suspensions of charcoal in phosphate buffer and in charcoal yeast extract medium readily decomposed exogenous peroxide (17 and 23 nmol/ml per min, respectively) . Combinations of bovine superoxide dismutase and catalase also decreased the rate of photooxidation of YE medium . Medium protected from light did not accumulate appreciable levels of hydrogen peroxide, and autoclaved YE medium protected from light supported good growth of Legionella micdadei . Various species of Legionella (10(4) cells per ml) exhibited sensitivity to relatively low levels of hydrogen peroxide (26.5 microM) in challenge experiments . The level of hydrogen peroxide that accumulated in YE medium over a period of several hours (greater than 50 microM) was in excess of the level tolerated by Legionella pneumophila, which contained no measurable catalase activity . Strains of L . micdadei, Legionella dumoffi, and Legionella bozmanii contained this enzyme, but the presence of catalase did not appear to confer appreciable tolerance to exogenously generated hydrogen peroxide.

Infect Immun, 1983 Mar, 39(3), 1161 - 6
Studies on the catalase of Histoplasma capsulatum; Howard DH; Factors which control the levels of catalase within yeast cells of Histoplasma capsulatum were studied . Only a small fraction of the total catalase activity could be detected in whole cells . The bulk of the activity was revealed in cell-free extracts or in cells permeabilized with acetone . The formation of the enzyme was regulated by glucose and by oxygen . There were large, consistent differences in the levels of catalase among strains of H . capsulatum . The sensitivity of the strains to H2O2 toxicity also varied remarkably . Peroxidase activity could not be detected in cell-free extracts of the strains . Resistance to H2O2 did not correspond to levels of catalase . There was no obvious correlation of H2O2 sensitivity and virulence among the strains.

Biochem J, 1983 Mar 1, 209(3), 669 - 76
Partial purification and regulatory properties of phosphofructokinase from Aspergillus niger; Habison A et al.; Phosphofructokinase (EC 2.7.1.11) from a citric acid-producing strain of Aspergillus niger was partially purified by the application of affinity chromatography on Blue Dextran--Sepharose and the use of fructose 6-phosphate and glycerol as stabilizers in the working buffer . The resulting preparation was still impure, but free of enzyme activities interfering with kinetic investigations . Kinetic studies showed that the enzyme exhibits high co-operativity with fructose 6-phosphate, but shows Michaelis--Menten kinetics with ATP, which inhibits at concentrations higher than those for maximal activity . Citrate and phosphoenolpyruvate inhibit the enzyme; citrate increases the substrate (fructose 6-phosphate) concentration for half-maximal velocity, {S}0.5, and the Hill coefficient, h . The inhibition by citrate is counteracted by NH4+, AMP and phosphate . Among univalent cations tested only NH4+ activates by decreasing the {S}0.5 for fructose 6-phosphate and h, but has no effect on Vmax . AMP and ADP activate at low and inhibit at high concentrations of fructose 6-phosphate, thereby decreasing the {S}0.5 for fructose 6-phosphate . Phosphate has no effect in the absence of citrate . The results indicate that phosphofructokinase from A . niger is a distinct species of this enzyme, with some properties similar to those of the yeast enzyme and in some other properties resembling the mammalian enzyme . The results of determinations of activity at substrate and effector concentrations resembling the conditions that occur in vivo support the hypothesis that the apparent insensitivity of the enzyme to citrate during the accumulation of citric acid in the fungus is due to counteraction of citrate inhibition by NH4+.

J Immunol Methods, 1983 Feb 25, 57(1-3), 291 - 4
The stabilization of the C3b molecule in immune deposits by formalin; Hed J et al.; Formalin-fixed tissue may be used as substrate for immunohistochemistry after enzymatic treatment with proteases . However, whether the antigenicity of C3b is restored is controversial owing to the protease sensitivity of the native molecule . The antigenicity and protease sensitivity of formalin-fixed and non-fixed C3b were tested by using C3b-opsonized yeast particles and the immunofluorescence technique . The results indicate that formalin, probably due to cross-linking, stabilizes C3b molecules and renders them less susceptible to protease treatment . Failures to detect C3b in immune deposits in fixed tissues may be due to factors other than protease treatment.

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