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 phosp