Yeasts

Cell, 1989 Sep 8, 58(5), 833 - 46
Isolation of a human cyclin cDNA: evidence for cyclin mRNA and protein regulation in the cell cycle and for interaction with p34cdc2; Pines J et al.; This paper reports the nucleotide and predicted amino acid sequence of a human B-type cyclin . The predicted protein sequence shows strong homology to the other known cyclins in the central third of the protein . We show that the level of cyclin mRNA is regulated during the cell cycle, increasing during G2 phase to four time that present in G1 . The protein accumulates steadily during G2 to at least 20 times its level in G1 and is abruptly destroyed at mitosis . In G2/M phase, cyclin is associated with p34cdc2, the human homolog of the fission yeast gene cdc2+, and this complex has histone H1 kinase activity.

Nature, 1989 Sep 7, 341(6237), 74 - 6
Changing fos oncoprotein to a jun-independent DNA binding protein with GCN4 dimerization specificity by swapping "leucine zippers"; Sellers JW et al.; A structural motif for DNA-binding proteins, the 'leucine zipper', has been proposed for the jun, fos and myc gene products, the yeast transcriptional activator GCN4, and the C/EBP enhancer-binding protein . These proteins all contain a region with four or five leucine residues spaced exactly seven amino acid residues apart whose sequence is consistent with the formation of an amphipathic alpha-helix . It has been proposed that the leucine zipper consists of two interdigitated alpha-helices, one from each monomer, that constitute the dimerization function necessary for high-affinity binding to DNA; an adjacent region of basic residues is thought to be responsible for specific protein-DNA contacts . In support of this model, substitution of the leucine residues within the motif can abolish dimerization and DNA-binding, and a synthetic peptide corresponding to the GCN4 leucine zipper forms alpha-helical dimers . Despite the conserved leucine residues, however, each protein has a distinct dimerization specificity . Specifically, GCN4 homodimer, Jun homodimer and Fos-Jun heterodimer proteins bind to the same DNA site, whereas Fos is unable to form homodimers, bind DNA, or interact with GCN4 (refs 8-14) . Here, we alter the dimerization specificity of Fos by precisely replacing its leucine zipper with that from GCN4 . This Fos-GCN4 chimaeric protein is able to bind to the target site in the absence of Jun, and can form DNA-binding heterodimers with GCN4 but not with Jun . These results indicate that the leucine zipper is sufficient to confer dimerization specificity and strongly suggest that Fos contacts DNA directly.

Am J Med, 1989 Sep 4, 87(3A), 26S - 29S
Protective efficacy of a recombinant deoxyribonucleic acid hepatitis B vaccine in institutionalized mentally handicapped clients; Van Damme P et al.; Mentally handicapped clients in institutions are at high risk for hepatitis B virus (HBV) infection . In 1985, 770 mentally handicapped residents from four institutions in the Antwerp area were screened for HBV markers . The prevalence of hepatitis B surface antigen was 10.3 percent (range, 6.1 to 15.2 percent); 42.3 percent (range, 11.5 to 60.1 percent) had antibodies to hepatitis B surface antigen and the hepatitis B core antigen . In 1986, 275 seronegative mentally handicapped residents were vaccinated intramuscularly in the deltoid region with 20 micrograms (1.0 ml) of a recombinant deoxyribonucleic acid yeast-derived hepatitis B vaccine (Engerix-B, SmithKline Biologicals, Rixensart, Belgium) on a zero-, one-, six-month schedule . Serum samples were collected at Months 1, 2, 7, 12, and 24 and were tested for HBV markers by radioimmunoassay . The seroconversion rates for hepatitis B surface antigen antibodies were 39 percent at Month 1 (geometric mean concentration, 6.4 IU/liter), 82 percent at Month 2 (geometric mean concentration, 23.4 IU/liter), 97 percent at Month 7 (geometric mean concentration, 1,034 IU/liter), and 96 percent at Month 12 (geometric mean concentration, 269 IU/liter) . Among the 214 residents evaluated at Month 12, 69 percent had hepatitis B surface antigen antibody levels greater than 100 IU/liter (geometric mean concentration, 603 IU/liter) . No significant adverse reactions were observed . Within the first seven months of observation, HBV infection was detected in eight of 271 subjects (estimated annual incidence of 5 percent) . During this period, none of the clients developed clinical hepatitis or showed biochemical evidence of liver damage . Between eight and 24 months, no additional HBV infections were detected . These data can be compared with an annual incidence of HBV infection of 8.7 percent in a historical cohort of mentally handicapped residents in one of the four institutions.

Am J Vet Res, 1989 Sep, 50(9), 1481 - 5
Characterization of the attachment of Treponema hyodysenteriae to Henle intestinal epithelial cells in vitro; Bowden CA et al.; Properties of the attachment of Treponema hyodysenteriae to Henle intestinal epithelial (HIE 407) cells were examined . The frequency of attachment depended on the motility and viability of the spirochetes . Rabbit hyperimmune and swine convalescent antisera inhibited attachment . Treatment of HIE cells with neuraminidase had no effect on attachment; however, treatment of spirochetes with the enzyme decreased adherence significantly (P = 0.01) . Attachment was inhibited by N-acetylneuraminic acid, D-glucuronic acid, and fetuin . Adherence was increased following coincubation with N-acetylglucosamine or yeast mannan . Surface antigens of T hyodysenteriae, isolated by chemical extraction, competitively inhibited adherence . Concentrated T hyodysenteriae culture supernatant fractions inhibited adherence, but concentrated phosphate buffered-saline washings of the spirochete and concentrated uninoculated media did not inhibit adherence . Sialic acid was detected in unwashed T hyodysenteriae and spent culture supernatant fractions in higher concentrations than from washed spirochetes and uninoculated media . It was concluded that the binding adhesins on T hyodysenteriae for cultured HIE cells may contain sialic acid residues.

Mol Cell Biol, 1989 Sep, 9(9), 3614 - 20
Sequence identification of cytochrome b in Plasmodium gallinaceum; Aldritt SM et al.; We have identified a gene that encodes the polypeptide cytochrome b in the avian malarial parasite Plasmodium gallinaceum . The gene containing the open reading frame was found to be located on a 6.2-kilobase multimeric extrachromosomal element . The amino acid translation from this gene demonstrated significant similarities to cytochrome b sequences from yeast, mammal, and fungus genomes . We present evidence that the P . gallinaceum cytochrome b transcript is part of a larger primary transcript from the element that is subsequently processed . The message for P . gallinaceum cytochrome b was found to be 1.2 kilobases in size . This is the first report identifying a mitochondrial nucleic acid sequence in malaria-causing organisms and suggests that a functional cytochrome system may exist in these parasites.

J Clin Microbiol, 1989 Sep, 27(9), 2003 - 7
New fluorescence assay for the quantitation of fungi; Coleman T et al.; Quantitative determination of fungal mass is easily achieved with a new procedure that detects particle epifluorescence . Fungi are detected after exposure to a fluorescent stain (Fungiqual; CIBA-GEIGY Corp., Summit, N.J.) by using a fluorescence particle concentration analyzer . This report describes a simple fluorescence method for quantitation of either yeast or mycelial forms of fungi . The nature of the staining reaction was studied, and a practical application of this procedure for determination of fungal susceptibility to an antifungal agent is presented.

Proc Natl Acad Sci U S A, 1989 Sep, 86(17), 6773 - 7
Human platelet glycoprotein IX: an adhesive prototype of leucine-rich glycoproteins with flank-center-flank structures; Hickey MJ et al.; The glycoprotein (GP) Ib-IX complex on the surface of human platelets functions as the von Willebrand factor receptor and mediates von Willebrand factor-dependent platelet adhesion to blood vessels . GPIX is a relatively small (Mr, 17,000) protein that may provide for membrane insertion and orientation of the larger component of the complex, GPIb (Mr, 165,000) . Using antibody screening, we cloned a cDNA encoding GPIX from a human erythroleukemia cell cDNA library constructed in phage lambda gt11 . Lacking a 5' untranslated region and start codon, the cDNA sequence includes 604 nucleotides, beginning with 495 bases at the 5' end coding for 165 amino acids, followed by a stop codon and 106 noncoding bases at the 3' end . By Northern blot analysis, the GPIX cDNA hybridizes with a single 1.0-kilobase species of platelet poly(A)+ RNA . Translation of the cDNA sequence gives a predicted protein sequence beginning with a truncated putative signal sequence of 5 amino acid followed by a sequence of 17 amino acids matching that determined directly by Edman degradation of intact GPIX . The predicted amino acid sequence of mature GPIX includes an NH2-terminal extracytoplasmic domain of 134 residues, a transmembrane domain of 20 residues, 6 intracytoplasmic residues, and 1 N-linked glycosylation site . GPIX contains a leucine-rich glycoprotein (LRG) sequence of 24 amino acids similar to conserved LRG sequences in GPIb and other proteins from humans, Drosophila, and yeast . "Flanking" sequences of approximately 22 amino acids are present at the NH2 and/or COOH sides of the "central" LRG sequence(s) in GPIX, GPIb, and the other human and Drosophila members of the LRG family . The role of the flank-LRG center-flank structure in the evolution and function of the LRG proteins remains to be defined.

Proc Natl Acad Sci U S A, 1989 Sep, 86(17), 6686 - 90
Alu polymerase chain reaction: a method for rapid isolation of human-specific sequences from complex DNA sources; Nelson DL et al.; Current efforts to map the human genome are focused on individual chromosomes or smaller regions and frequently rely on the use of somatic cell hybrids . We report the application of the polymerase chain reaction to direct amplification of human DNA from hybrid cells containing regions of the human genome in rodent cell backgrounds using primers directed to the human Alu repeat element . We demonstrate Alu-directed amplification of a fragment of the human HPRT gene from both hybrid cell and cloned DNA and identify through sequence analysis the Alu repeats involved in this amplification . We also demonstrate the application of this technique to identify the chromosomal locations of large fragments of the human X chromosome cloned in a yeast artificial chromosome and the general applicability of the method to the preparation of DNA probes from cloned human sequences . The technique allows rapid gene mapping and provides a simple method for the isolation and analysis of specific chromosomal regions.

Indian J Exp Biol, 1989 Sep, 27(9), 785 - 91
Development of improved media for axenic cultivation of Acanthamoeba culbertsoni, Singh and Das 1970; Shukla OP et al.; Several varieties of peptone supported growth of A . culbertsoni to different extents reaching a maximum cell density of 1-2 X 10(6)/ml . Proteose peptone and tryptone also yielded good growth when combined with thiamine and vitamin B12 . A combination of proteose peptone with glucose, yeast extract and salts promoted excellent growth of A . culbertsoni with cell density reaching 1-2 X 10(7) cells/ml; tryptone and one of the indigenous peptones also yielded comparable growth when substituted for proteose peptone in this medium . Casamino acids also supported good growth of amoebae and requirement of yeast extract could be met by a combination of thiamine, vitamin B12 and biotin . Bacto peptone did not support good growth of this amoeba but supplementation of peptone with casamino acids or amino acid mixture improved the growth supporting capacity of the medium . Development of several media with or without glucose will aid in cultivation of A . culbertsoni, studies on its metabolism as well as screening of potential drugs.

Mycoses, 1989 Sep, 32(9), 463 - 8
Candida guilliermondii var . guilliermondii infection in infertile women; Nagy B et al.; The sera of 263 women--217 infertile and 46 pregnant--were examined by various serological methods (precipitation test, agglutination, indirect immunofluorescence) to detect Candida guilliermondii var . guilliermondii (C.g.) infection . The precipitation reaction was performed with extracellular C . guilliermondii antigen, the agglutination reaction was employed parallel with C . albicans . In the infertile group 122 (56.2%) proved to be C.g . positive, while in the fertile 11 women (23.9%) proved to be so, the level of significance being p less than 0.0001 between the two groups . A one-month ketoconazole treatment (one tablet, 200 mg/day) was adequate for eliminating the C.g . infection . In a few cases hystological examinations were also performed according to Gomori-Grocott and yeast cells could be detected in the stroma of the ovary . IgA, IgG, IgM, Gc-globulin, transferrin and ferritin determinations were carried out before and after the ketoconazole treatment, and there were significant differences in the IgM and transferrin levels between the infected and non-infected groups . The authors achieved 5 pregnancies of 56 treated women in 6 months.

Trends Biochem Sci, 1989 Sep, 14(9), 368 - 73
Quantum mechanical effects in enzyme-catalysed hydrogen transfer reactions; Klinman JP; Hydrogen tunneling at room temperature has recently been demonstrated in the reactions catalysed by yeast alcohol dehydrogenase and bovine serum amine oxidase . These results suggest that quantum mechanical effects may be a fundamental and pervasive feature of enzyme-catalysed hydrogen transfer reactions . Our ability to detect such behavior introduces a new probe for the investigation of reaction barrier shape and protein dynamics in enzyme catalysis.

G Ital Dermatol Venereol, 1989 Sep, 124(9), XLVII - IL
{Efficacy of fenticonazole in patients with pityriasis versicolor}; Di Silverio A et al.; Thirty patients suffering from Pityriasis versicolor were treated with fenticonazole in cream or lotion form with two applications a day . At microscopy, Malassezia furfur was encountered in 25 cases and Pityrosporum orbiculare in 5 . Disappearance of the yeast was obtained on average in 2 weeks of treatment . Measurement of sebum content with a Sebumeter apparatus did not reveal significant difference (p greater than 0.05) between patients suffering from Pityriasis versicolor and controls and in treated patients, in the course of topical therapy.

Arch Pharm (Weinheim), 1989 Sep, 322(9), 531 - 4
Antifungal activity of novel 5-carbonyl derivatives of 3-phenyl-3-(1H-imidazol-1-ylmethyl)-2-methylisoxazolidines; Bennett GA et al.; The synthesis and antifungal activity of a series of novel 5-carbonyl derivatives of 3-phenyl-3-(1H-imidazol-1-ylmethyl)-2-methylisoxazolidines (4) are discussed . The preparation of the title compounds involved a 1,3-dipolar cycloaddition reaction of alpha-substituted ketonitrones with either acrylic esters, acrylamide or methyl vinyl ketone to furnish cis/trans-diastereomeric mixtures of the desired 5-carbonyl isoxazolidines 4 . The anifungal activity was evaluated in vitro in solid agar cultures . Some of the compounds tested exerted moderate to potent activity against a wide variety of dermatophytes and yeast and systemic fungi.

Am J Gastroenterol, 1989 Sep, 84(9), 1079 - 83
Fungal colonization of the esophagus; Vermeersch B et al.; A prospective study was conducted in 224 patients to determine the clinical significance of esophageal colonization with yeasts under different conditions . In accordance with the results of direct smear microscopic examination and culture of esophageal brushings, patients were divided into three groups: positive, negative, and the patients, in whom saprophytic forms were detected . A higher prevalence of positive findings was noted in patients with predisposing factors for yeast invasion than in patients free of underlying disease . Eleven percent of patients with an endoscopically normal appearing esophagus were positive . We have the impression that this situation may represent a preclinical condition of fungal esophagitis . Patients treated with H2-blocking agents showed a significantly higher incidence of positive findings, than did those without such treatment . Whether patients suffering from a refluxesophagitis resistant to long-term treatment with H2 blockers, but with a significant colonization by yeasts, could benefit by an additional treatment with antimycotics remains a controversial issue and should be studied in a controlled way.

J Biochem (Tokyo), 1989 Sep, 106(3), 396 - 400
N-terminal half of a mitochondrial presequence peptide takes a helical conformation when bound to dodecylphosphocholine micelles: a proton nuclear magnetic resonance study; Endo T et al.; Two-dimensional proton nuclear magnetic resonance (NMR) spectra of a synthetic peptide (p25) corresponding to the amino-terminus of the yeast mitochondrial cytochrome oxidase subunit IV precursor protein have been analyzed . Sequence-specific resonance assignments of the peptide have been made in the presence of micelles of a phospholipid analog, perdeuterated dodecylphosphocholine (DPC), with the aid of such techniques as HOHAHA, DQF-COSY, and NOESY . The interresidue nuclear Overhauser effects (NOEs) indicate that the N-terminal half of p25 (S3-F11) takes a helical structure while the C-terminal half does not take a regular secondary structure . Addition of DPC to the solution of p25 induced chemical shift changes only of the resonances from the residues in the N-terminal half, suggesting that the N-terminal half of p25 is directly involved in binding to DPC . The induced helical structure in the N-terminal half at a lipid-water interface may be important in the ability of this presequence to direct a "passenger" protein into mitochondria.

J Med Entomol, 1989 Sep, 26(5), 487 - 8
Larval diet and the vector competence of Culex annulirostris (Diptera: Culicidae) for Murray Valley encephalitis virus; Kay BH et al.; Culex annulirostris Skuse larvae were reared on two different amounts of powdered dog chow and yeast, low intake (2.8 micrograms/larva) and high intake (8.4 micrograms/larva), to produce adults with mean wing lengths of 3.25 and 3.7 mm, respectively . The resulting adults were fed different dosages of Murray Valley encephalitis virus and after 10 d extrinsic incubation at 28 degrees C, the infection rate, transmission rate, salivary gland, and body titers were determined . There were no significant differences in any of these parameters.

Anal Biochem, 1989 Sep, 181(2), 227 - 33
Computer-controlled discontinuous rotating gel electrophoresis for separation of very large DNA molecules; Gekeler V et al.; The newly designed equipment for alternating field gel electrophoresis which permits the separation of very large DNA molecules and the simultaneous analysis of up to 35 samples is described . The field alternation is effected by intermittently rotating the submerged agarose gel by optitional angles . The time intervals between changes of position are controlled by a computer program driving a simple switching device which was designed to suit any technique using periodic switching or inversion of the electrical field . Because the electrophoresis unit provides an absolutely homogeneous electrical field, no distortion of migration lanes occurs and the resolution is very good . Moreover, by using a switching time interval gradient an almost perfect linear relationship between migration distances and molecule sizes in the range of about 100-1250 kilobase pairs is observed . In two-dimensional separations, different switching time programs for the first and second dimension allow maximum resolution of selected size ranges . Field inversion gel electrophoresis is possible as well . The performance of the method is demonstrated by comparing the chromosome sizes of different yeast strains.

J Clin Invest, 1989 Sep, 84(3), 886 - 91
Receptor-mediated phagocytosis in human neutrophils is associated with increased formation of inositol phosphates and diacylglycerol . Elevation in cytosolic free calcium and formation of inositol phosphates can be dissociated from accumulation of diacylglycerol; Fallman M et al.; Phagocytosis of C3bi- or IgG-opsonized yeast particles in human neutrophils was found to be associated with an increased formation of inositol phosphates and diacylglycerol . Pertussis toxin only marginally affected phagocytosis of IgG- and C3bi-opsonized particles and the associated formation of second messengers . Forskolin, which induced a threefold rise of cellular cAMP, however, markedly inhibited both C3bi- and IgG-mediated phagocytosis as well as the particle-induced formation of inositol phosphates and diacylglycerol . These observations are in contrast to what was found to occur with chemotactic factors and indicate that chemotactic and phagocytic signaling can be regulated independently in human neutrophils . Since C3bi-mediated phagocytosis has been shown to occur at vanishingly low cytosolic free calcium levels, calcium-depleted cells were used to study the importance of the inositol cycle for the engulfment of C3bi-opsonized particles . Despite a total lack of receptor-induced formation of inositol phosphates, a significantly increased accumulation of diacylglycerol accompanied the ingestion of C3bi-opsonized particles . These data show that the engulfment of C3bi-opsonized particles can occur independently of both a calcium transient and an increased inositol phosphate production . However, the observed accumulation of diacylglycerol, not derived from phosphoinositides, suggests that this second messenger play a role in the control of the engulfment process.

Br Poult Sci, 1989 Sep, 30(3), 633 - 9
Utilisation of free and protein dietary lysine in chicks estimated with isotopes; Otto M et al.; 1 . Utilisation of supplementary free L-lysine hydrochloride was estimated in growing chicks and compared to that of protein-bound lysine . The technique used is based on the oxidative catabolism of either free (U-14C)-L-lysine or the labelled lysine incorporated into yeast proteins . 2 . The animals received a wheat-wheat gluten diet which was L-lysine-supplemented with either unlabelled yeast proteins or a mixture of synthetic amino acids simulating the yeast proteins or L-lysine hydrochloride alone . 3 . At 13 and 15 days after hatching, expiry of (14C)--carbon dioxide was followed 8 h after dosing with the appropriate radiolabelled diet . After 4 h, 0.66% of the protein-bound and 5.3 to 5.7% of the free lysine radioactivity appeared as 14C--carbon dioxide . 4 . It is concluded that under these conditions lysine from both sources was utilised more efficiently than had been assumed hitherto, protein-bound lysine being slightly better utilised than free lysine.

J Mol Evol, 1989 Sep, 29(3), 202 - 7
Evolution of the mitochondrial genetic code . I . Origin of AGR serine and stop codons in metazoan mitochondria; Osawa S et al.; AGA and AGG (AGR) are arginine codons in the universal genetic code . These codons are read as serine or are used as stop codons in metazoan mitochondria . The arginine residues coded by AGR in yeast or Trypanosoma are coded by arginine CGN throughout metazoan mitochondria . AGR serine sites in metazoan mitochondria are occupied mainly in corresponding sites in yeast or Trypanosoma mitochondria by UCN serine, AGY serine, or codons for amino acids other than serine or arginine . Based on these observations, we propose the following evolutionary events . AGR codons became unassigned because of deletion of tRNA Arg (UCU) and elimination of AGR codons by conversion to CGN arginine codons . Upon acquisition by serine tRNA of pairing ability with AGR codons, some codons for amino acids other than arginine mutated to AGR, and were captured by anticodon GCU in serine tRNA . During vertebrate mitochondrial evolution, AGR stop codons presumably were created from UAG stop by deletion of the first nucleotide U and by use of R as the third nucleotide that had existed next to the ancestral UAG stop.

J Neurogenet, 1989 Sep, 6(1), 11 - 26
The expression of the neurogenic locus Notch during the postembryonic development of Drosophila melanogaster and its relationship to mitotic activity; Markopoulou K et al.; The molecular analysis of the Notch locus of Drosophila melanogaster demonstrated that it codes for a protein which shows homology to the epidermal growth factor as well as to the products of certain yeast genes involved in the control of the cell cycle (Wharton et al., 1985a; Breeden and Nasmyth, 1987) . The structure of the protein suggests that Notch is involved in a cell interaction mechanism which controls the differentiation of several different tissues during development . Here we examine Notch expression during imaginal development using in situ hybridization to tissue sections and demonstrate that Notch is not expressed ubiquitously during the postembryonic stages, but rather is confined to specific tissues . During the larval and early pupal period Notch transcripts are predominantly localized in the imaginal discs and the central nervous system . In the middle and late pupal period the signal levels in these tissues drop dramatically and in the adult animal Notch transcripts are essentially detected only in the ovaries . In the larval stages the pattern of Notch expression appears to be closely correlated with mitotically active tissues, while in later stages this correlation appears less strict . The findings reported here indicate that there is a requirement for normal Notch function in a number of tissues at several developmental stages and that the pleiotropic phenotypic manifestation of Notch mutations is a context dependent developmental result . The observed association of Notch expression with mitotically active cell populations raises the possibility that Notch may play a role in the cell cycle.

Jpn J Cancer Res, 1989 Sep, 80(9), 879 - 86
The tumor rejection antigen separated from Rous sarcoma virus-induced murine fibrosarcoma exhibits a molecular weight of approximately 60 kD but differs from functional pp60src; Suda T et al.; The tumor antigen capable of inducing tumor resistance (tumor rejection antigen; TRA) was obtained in a solubilized form by sodium dodecyl sulfate (SDS) extraction of plasma membrane fraction from Rous sarcoma virus (RSV)-induced CSA1M fibrosarcoma cells (BALB/c origin) . Analyses by Sephacryl S-300 gel filtration and SDS-polyacrylamide gel electrophoresis revealed that TRA activity was recovered in the fraction with a molecular weight of approximately 60 kD . Unfractionated crude SDS-solubilized preparation contained gp70 as detected by rabbit anti-gp70 antiserum, whereas such reactivity was lost in the fraction exhibiting the molecular weight of about 60 kD . Since this fraction retained pp60src activity, the relation of TRA to pp60src was further investigated . pp60v-src was also obtained from the lysate of v-src-expressing yeast transformant . Immunization of BALB/c mice with such pp60v-src-containing lysate failed to induce any significant tumor protection . The above 60 kD fraction of CSA1M solubilized antigens was allowed to bind to Sepharose beads coupled with anti-pp60src monoclonal antibody and separated into the bead-bound and bead-unbound fractions . The bead-bound fraction that was recovered from pp60src-binding beads (pp60src-positive fraction) did not exhibit the TRA activity . In contrast, immunization with the fraction depleted of pp60src activity (bead-unbound fraction) resulted in potent tumor protection . These results indicate that the solubilized membranous component(s) of CSA1M with a molecular weight of approximately 60 kD, which is distinct from functional pp60src, functions as the TRA against RSV-induced CSA1M tumor cells.

Biochim Biophys Acta, 1989 Aug 31, 997(3), 236 - 41
Nonpolar interactions in the modification of an essential sulfhydryl of sorbitol dehydrogenase by N-alkylmaleimides; Beier KH et al.; A series of N-alkylmaleimides varying in chainlength from N-methyl- to N-octylmaleimide inclusive was shown to effectively inactivate sheep liver sorbitol dehydrogenase at pH 7.5 and 25 degrees C . The apparent second-order rate constants for inactivation increased with increasing chainlength of the N-alkylmaleimide used . Positive chainlength effects were also indicated by the Kd values for the N-ethyl and N-heptyl derivatives obtained from studies of the saturation kinetics observed for inactivation of the enzyme at high concentrations of these maleimides . The complete inactivation of sorbitol dehydrogenase was demonstrated to occur through the selective covalent modification of one cysteine residue per subunit of enzyme . The stoichiometry of enzyme inactivation was supported on the one hand by fluorescence titration with fluorescein mercuric acetate of the native and the inactivated enzyme, and, on the other hand, by the simultaneous inactivation of the enzyme with selective modification of one sulfhydryl per subunit by N-{p-(2-benzoxazolyl)phenyl}maleimide . Protection of the enzyme from N-alkylmaleimide inactivation was observed with the binding of NADH, whereas both NAD and sorbitol were ineffective as protecting ligands . Diazotized 3-aminopyridine adenine dinucleotide, in contrast to previous studies of this reagent with yeast alcohol dehydrogenase and rabbit muscle glycerophosphate dehydrogenase, did not function as a site-labeling reagent for sorbitol dehydrogenase.

Wien Med Wochenschr, 1989 Aug 31, 139(15-16), 381 - 3
{Prevention of candidiasis in at-risk patients}; Hain K; Colonisation by yeasts and invasive yeast-growth (candidosis, yeast-mycosis) increase whenever a patient's power of resistance is lowered . Early recognition of colonizing yeasts in a patient can be as important for his vital functions, as the therapy of the underlying disease will be . This seems to be quite complicated in many cases and often it is omitted by several reasons . An "easy to handle method" is a "rapid yeast culture" . Blood-cultures and serodiagnosis can be added if held necessary . It is more important to find out, if there are few or many yeast-cells in a patient, than to count them . Hidden "yeast-nests" can in short time recruit eliminated yeasts . Too much sugar means too many yeasts as well . Anti-yeast-therapeutics have to be selected and dosed according to their characteristic qualities.

Wien Med Wochenschr, 1989 Aug 31, 139(15-16), 379 - 80
{Dermatomycoses and an antifungal diet}; Putzier E; Treatment of dermatomycoses and some other dermatoses, e.g . seborrheic eczema, is more effective if the intestinal tract is free of pathogenic fungi . In order to reduce the yeast colonies between the intestinal villi and to avoid the persorption of living yeast cells into the lymph-tract and the blood circulation it is recommendable to perform the so-called antifungal diet: less sugar of all kind and much more vegetable fibres in the food.

J Biol Chem, 1989 Aug 25, 264(24), 14240 - 5
Proalbumin to albumin conversion by a proinsulin processing endopeptidase of insulin secretory granules; Rhodes CJ et al.; A lysate of purified insulin secretory granules, which contains two types of proinsulin processing activity (type 1, Arg-Arg-directed and type II, Lys-Arg-directed (Davidson, H.W., Rhodes, C.J., and Hutton, J . C . (1988) Nature 333, 93-96), was found to process proalbumin by specific proteolytic cleavage of the COOH-terminal side of the Arg-2-Arg-1 sequence . The subcellular distribution of proalbumin processing activity in insulinoma tissue paralleled that for proinsulin conversion and occurred principally in a secretory granule fraction . Cleavage appeared to result from the Arg-Arg-directed type 1 proinsulin processing endo-peptidase . It was Ca2+-dependent (K0.5 activation = 1.0-1.5 mM Ca2+), unaffected by group-specific inhibitors of serine, cysteinyl, or aspartyl proteinases, and had an acidic pH optimum (5.5) . Active-site inhibitor studies showed this activity had a preference for dibasic over monobasic amino acid sequences and indicated that the sequence of the dibasic site was an important determinant of the susceptibility of the substrate to cleavage . The activity did not process the proalbumin Christchurch mutant (Arg-2-Arg-1 to Arg-2-Gln-1) . It was inhibited by the variant alpha 1-antitrypsin Pittsburgh (Met358 to Arg358; K0.5 = 100 nM) but not by other related proteins normally co-secreted with albumin from hepatocytes, namely alpha 1-antitrypsin M, alpha 2-macroglobulin, or antithrombin III . The insulin secretory granule proalbumin processing activity was indistinguishable from a proalbumin endopeptidase reported in rat liver membranes and similar to the yeast KEX-2 protease . These findings suggest that a highly conserved set of proprotein endopeptidases exists, which are specific for a dibasic sequence but broadly specific for proprotein substrates . Such enzymic activities appear to be active within both the constitutive and regulated pathways of secretion . Intraorganellar Ca2+ and pH appear to play a key role in regulating their activities.

J Mol Biol, 1989 Aug 20, 208(4), 587 - 99
Processing of mitochondrial RNA in Aspergillus nidulans; Dyson NJ et al.; Genes for cytochrome oxidase subunit I (oxiA), ATPase subunit 9, NADH dehydrogenase subunit 3 (ndhC) and cytochrome oxidase subunit II (oxiB) are located within a 7.2 kb (1 kb = 10(3) bases or base-pairs) segment of the Aspergillus nidulans mitochondrial genome . Northern hybridization shows that abundant RNA molecules of 4.0, 2.5 and 1.5 kb, each containing copies of two or more genes, are transcribed from this region . The 4.0 kb molecule, which contains copies of each of the four genes but lacks the three oxiA introns, is cleaved at a point just upstream from ndhC to give rise to the 2.5 kb RNA, which contains copies of oxiA and the ATPase subunit 9 gene, and the 1.5 kb RNA, which carries ndhC and oxiB . The ATPase subunit 9 gene, which has no identified function, is therefore transcribed into an abundant RNA . S1 nuclease analysis indicates that there are no additional introns in the amino-terminal region of oxiA and that the 4.0 and 2.5 kb transcripts of this gene have staggered 5' termini, the most upstream of which is adjacent to the 3' end of the histidinyl-tRNA gene . The results suggest that transcription of this genome proceeds via a very limited number of primary transcripts with mature RNAs produced by extensive processing events including tRNA excision . RNA synthesis and processing in A . nidulans mitochondria therefore resembles the events occurring in metazoa rather than yeast.

Nature, 1989 Aug 17, 340(6234), 568 - 71
Leucine zippers of fos, jun and GCN4 dictate dimerization specificity and thereby control DNA binding; Kouzarides T et al.; The products of the fos and jun protooncogenes form a stable heterodimer which binds to the TPA-responsive element (TRE) TGACTCA with high affinity . These two proteins, together with the yeast GCN4 protein, belong to a growing family of transcription factors, including FosB, Fra1, JunB and JunD, whose members share a highly conserved DNA-binding domain . This domain is composed of two structures: a basic motif, which is thought to bind directly to DNA; and a leucine zipper, which provides a dimerization interface . Although this domain is highly conserved in Fos, Jun and GCN4, each of these three proteins has very different relative affinities for the TRE . To understand these differences, we used 'domain-swapping' experiments designed to test the relative contributions of the basic motif and the leucine zipper to TRE-binding affinity . Here we show that fos, jun and GCN4 have different affinities for the TRE due to differences in the hetero- or homo-dimerization capacity of their leucine zipper domains; the basic motifs of these three proteins have comparable DNA binding potential . These results indicate that leucine zippers control the types of protein complexes which can associate with a TRE and regulate gene expression.

Mol Cell Biochem, 1989 Aug 15, 89(1), 57 - 67
Phosphoglucoisomerase-catalyzed interconversion of hexose-phosphates . Isotopic discrimination between hydrogen and tritium; Malaisse-Lagae F et al.; The rate of conversion of D-glucose 6-phosphate to D-fructose 6-phosphate as catalyzed by yeast phosphoglucoisomerase is about fourfold lower when 3H, rather than 1H, is present on the C2 of D-glucose 6-phosphate . This difference appears to be due mainly to a change in maximal velocity, rather than affinity . Phosphoglucoisomerase also distinguishes between 1H and 3H in terms of either their intramolecular transfer from C2 to C1 or their incorporation from water on the C1 of D-fructose 6-phosphate.

J Biol Chem, 1989 Aug 15, 264(23), 13929 - 37
Two genes encode the major membrane-associated protein of methanol-induced peroxisomes from Candida boidinii; Garrard LJ et al.; A massive proliferation of peroxisomes occurs in the yeast Candida boidinii when methanol is utilized as the sole carbon source; these peroxisomes contain the enzymes which catalyze the initial steps of methanol utilization . The most abundant peroxisomal membrane-associated protein has an apparent molecular mass of 20 kDa and is termed PMP20 . We report the isolation of two genes that encode very similar forms of PMP20; this is the first report of genes that encode proteins associated with peroxisomal membranes . Southern analysis demonstrates that the two genes are on different loci, although there are several homologous regions of both 5'- and 3'-untranslated sequence . One of the areas of 5' homology is within the untranslated region of the mRNA . Within the coding region there are 35 base differences between the two genes that are reflected in only five amino acid differences . The mRNAs representing both genes of PMP20 are induced in cells grown in methanol-containing medium and are below detection in cells grown in glucose . S1 nuclease protection analysis indicates that there is a 2.5-fold difference in mRNA expression between the two genes when induced . The predicted sequences of both PMP20 genes show the absence of a cleaved amino-terminal leader sequence and the presence of only 1 cysteine residue . In agreement with previous biochemical data suggesting a peripheral association of this protein with the membrane (Goodman, J . M., Maher, J., Silver, P . A., Pacifico, A., and Sanders, D . (1986) J . Biol . Chem . 261, 3464-3468), there are no obvious membrane spanning regions predicted in the sequences . Both PMP20 gene products contain the carboxyl-terminal sequence AKL, similar to the putative SKL peroxisomal sorting sequence (Gould, S . J., Keller, G.-A., and Subramani, S . (1988) J . Cell Biol . 107, 897-905).

Eur J Biochem, 1989 Aug 15, 183(3), 661 - 9
Purification and characterization of trimming glucosidase I from pig liver; Bause E et al.; Trimming glucosidase I has been purified about 400-fold from pig liver crude microsomes by fractional salt/detergent extraction, affinity chromatography and poly(ethylene glycol) precipitation . The purified enzyme has an apparent molecular mass of 85 kDa, and is an N-glycoprotein as shown by its binding to concanavalin A-Sepharose and its susceptibility to endo-beta-N-acetylglucosaminidase (endo H) . The native form of glucosidase I is unusually resistant to non-specific proteolysis . The enzyme can, however, be cleaved at high, that is equimolar, concentrations of trypsin into a defined and enzymatically active mixture of protein fragments with molecular mass of 69 kDa, 45 kDa and 29 kDa, indicating that it is composed of distinct protein domains . The two larger tryptic fragments can be converted by endo H to 66 kDa and 42 kDa polypeptides, suggesting that glucosidase I contains one N-linked high-mannose sugar chain . Purified pig liver glucosidase I hydrolyzes specifically the terminal alpha 1-2-linked glucose residue from natural Glc3-Man9-GlcNAc2, but is inactive towards Glc2-Man9-GlcNAc2 or nitrophenyl-/methyl-umbelliferyl-alpha-glucosides . The enzyme displays a pH optimum close to 6.4, does not require metal ions for activity and is strongly inhibited by 1-deoxynojirimycin (Ki approximately 2.1 microM), N,N-dimethyl-1-deoxynojirimycin (Ki approximately 0.5 microM) and N-(5-carboxypentyl)-1-deoxynojirimycin (Ki approximately 0.45 microM), thus closely resembling calf liver and yeast glucosidase I . Polyclonal antibodies raised against denatured pig liver glucosidase I, were found to recognize specifically the 85 kDa enzyme protein in Western blots of crude pig liver microsomes . This antibody also detected proteins of similar size in crude microsomal preparations from calf and human liver, calf kidney and intestine, indicating that the enzymes from these cells have in common one or more antigenic determinants . The antibody failed to cross-react with the enzyme from chicken liver, yeast and Volvox carteri under similar experimental conditions, pointing to a lack of sufficient similarity to convey cross-reactivity.

J Biol Chem, 1989 Aug 15, 264(23), 13697 - 700
Regulation of the product of a possible human cell cycle control gene CDC2Hs in B-cells by alpha-interferon and phorbol ester; Thomas NS; The product of the cell cycle control gene cdc2 is required in yeast for transition through both G1 and G2 control points of the cell cycle . The homologous protein in higher eukaryotes has been shown to be a component of the mitosis promoting factor complex and may thus regulate entry through the G2 control point into mitosis . It is suggested from the work presented here that, as in yeast, the human CDC2Hs gene product (p34CDC2Hs) may also play a role in cell cycle control in the G1(G0) phase of the cell cycle . Interferon-alpha inhibits the growth of the human B-cell line Daudi in the G1(G0) phase of the cell cycle and prevents cells from entering S-phase . Culturing the cells with interferon-alpha inhibits the phosphorylation of p34CDC2Hs and causes the down-regulation of CDC2Hs mRNA . Phorbol ester also inhibits the Daudi cell cycle in G1(G0) and causes the inhibition of p34CDC2Hs phosphorylation and a reduction of CDC2Hs mRNA . These studies provide insights into the process of growth control and the cytostatic mechanism of interferon-alpha.

Cell, 1989 Aug 11, 58(3), 553 - 63
glp-1 and lin-12, genes implicated in distinct cell-cell interactions in C . elegans, encode similar transmembrane proteins; Yochem J et al.; Genomic DNA closely related in sequence to lin-12, a gene that specifies certain cell fates during C . elegans development, was isolated from a C . elegans library by low stringency hybridization . DNA sequencing of genomic and cDNA clones predicts the new sequence to encode an integral membrane protein that shares three repeated amino acid sequence motifs with the lin-12 product and the Drosophila Notch product: an epidermal growth factor-like motif, the "lin-12/Notch Repeat," and a motif present in two yeast gene products that have cell cycle dependent functions . Austin and Kimble (see accompanying paper) present evidence that this sequence corresponds to glp-1, a gene implicated in cell-cell interactions distinct from those involving lin-12 . Possible implications of the predicted structure of the glp-1 product with respect to these cell-cell interactions are discussed.

Cell, 1989 Aug 11, 58(3), 473 - 83
The AU-rich sequences present in the introns of plant nuclear pre-mRNAs are required for splicing; Goodall GJ et al.; Plant cells do not in general process the introns of transcripts expressed from introduced vertebrate genes . By studying the processing of model introns in transfected plant protoplasts, we have investigated the special requirements for intron recognition by plant cells . Our results indicate that the requirements for intron recognition in plants are different from those of both metazoa and yeast . A synthetic intron of arbitrary sequence but incorporating splice site consensus sequences and a high proportion of U and A nucleotides, a characteristic feature of plant introns, was efficiently spliced in protoplasts . We have studied the effects of various sequence alterations and conclude that AU-rich sequences are necessary for intron recognition . In addition, we find that the criteria for branch site selection are relaxed, as they are in vertebrates, but a polypyrimidine tract is not necessary.

Nature, 1989 Aug 10, 340(6233), 487 - 8
Transcription factor IIIA induced bending of the Xenopus somatic 5S gene promoter; Schroth GP et al.; Transcription factor IIIA (TFIIIA), the canonical zinc-finger protein, is a protein of relative molecular mass 39,000 (39K) that is required for transcription of 5S-ribosomal subunit genes in Xenopus . It binds in a sequence-specific manner to the internal control region of the 5S gene (see Fig . 1) and facilitates transcription of the gene by RNA polymerase III . It also binds to the 5S gene product to form a 7S ribonucleoprotein particle . In oocytes the 7S particle acts as a storage form of the RNA to be utilized later in development . TFIIIA binds to DNA through its 30 K N-terminal domain, which contains nine zinc-fingers . TFIIIA was the first protein described to have this type of DNA binding motif, but numerous other proteins have now been shown to have zinc-finger domains . A structure for a single zinc-finger from the yeast protein ADR1, was recently proposed based on two-dimensional NMR data (ref . 8), and a similar structure was proposed based on comparison with crystal structures of other metalloproteins . Although models for the interaction of TFIIIA with the 5S-ribosomal gene DNA have been proposed, based on nuclease digestion and methylation interference data, little precise structural information is available for TFIIIA and the physical basis for the interaction of zinc-fingers with DNA is not understood . Using both circular permutation and circularization assays we provide convincing biochemical evidence that TFIIIA bends the DNA at the internal promoter of the 5S gene.

J Chromatogr, 1989 Aug 4, 476, 205 - 25
High-performance liquid chromatography of amino acids, peptides and proteins . XCII Thermodynamic and kinetic investigations on rigid and soft affinity gels with varying particle and pore sizes; Anspach FB et al.; In these investigations Cibacron Blue F3GA was immobilized on soft gels, porous silicas, and non-porous glass beads . Hen egg white lysozyme, human serum albumin and yeast alcohol dehydrogenase were used as adsorbates with the dye-affinity sorbents . Batch experiments with continuous monitoring of protein concentration were employed to evaluate thermodynamic and kinetic behaviour of these proteins in finite bath systems . The observed adsorption kinetic rates of interaction of the above proteins with each of the dye-affinity sorbents were found to decrease with increasing protein molecular weight . Equilibration times, in the batch experimental mode, of the adsorption of lysozyme on the dye-affinity sorbents varied from 20 s for the non-porous glass beads with a size range of 20-40 microns to more than 60 min in the case of a porous sorbent with a particle diameter of 100-300 microns and 60 nm pore size . Furthermore, equilibration times, which represent the overall adsorption rates incorporating all the non-equilibrium effects, increased with all affinity systems when adsorption took place in the non-linear portion of the isotherm . The most dramatic increase was observed when sorbents with relatively high protein size to pore size ratios, lambda, were employed.

Biochim Biophys Acta, 1989 Aug 3, 975(3), 377 - 83
Inhibition of the bovine-heart mitochondrial F1-ATPase by cationic dyes and amphipathic peptides; Bullough DA et al.; The bovine heart mitochondrial F1-ATPase is inhibited by a number of amphiphilic cations . The order of effectiveness of non-peptidyl inhibitors examined as assessed by the concentration estimated to produce 50% inhibition (I0.5) of the enzyme at pH 8.0 is: dequalinium (8 microM), rhodamine 6G (10 microM), malachite green (14 microM), rosaniline (15 microM) greater than acridine orange (180 microM) greater than rhodamine 123 (270 microM) greater than rhodamine B (475 microM), coriphosphine (480 microM) greater than safranin O (1140 microM) greater than pyronin Y (1650 microM) greater than Nile blue A (greater than 2000 microM) . The ATPase activity was also inhibited by the following cationic, amphiphilic peptides: the bee venom peptide, melittin; a synthetic peptide corresponding to the presence of yeast cytochrome oxidase subunit IV (WT), and amphiphilic, synthetic peptides which have been shown (Roise, D., Franziska, T., Horvath, S.J., Tomich, J.M., Richards, J.H., Allison, D.S . and Schatz, G . (1988) EMBO J . 7, 649-653) to function in mitochondrial import when attached to dihydrofolate reductase (delta 11.12, Syn-A2, and Syn-C) . The order of effectiveness of the peptide inhibitors as assessed by I0.5 values is: Syn-A2 (40 nM), Syn-C (54 nM) greater than melittin (5 microM) greater than WT (16 microM) greater than delta 11,12 (29 microM) . Rhodamines B and 123, dequalinium, melittin, and Syn-A2 showed noncompetitive inhibition, whereas each of the other inhibitors examined (rhodamine 6G, rosaniline, malachite green, coriphosphine, acridine orange, and-Syn-C) showed mixed inhibition . Replots of slopes and intercepts from Lineweaver-Burk plots obtained for dequalinium were hyperbolic indicating partial inhibition . With the exception of Syn-C, for which the slope replot was hyperbolic and the intercept replot was parabolic, steady-state kinetic analyses indicated that inhibition by the other inhibitors was complete . The inhibition constants obtained by steady-state kinetic analyses were in agreement with the I0.5 values estimated for each inhibitor examined . Rhodamine 6G, rosaniline, dequalinium, melittin, Syn-A2, and Syn-C were observed to protect F1 against inactivation by the aziridinium of quinacrine mustard in accord with their experimentally determined I0.5 values.(ABSTRACT TRUNCATED AT 400 WORDS)

Blut, 1989 Aug, 59(2), 165 - 70
Lineage- and differentiation-dependent alterations in the expression of receptors for glycoconjugates (lectins) in different human hematopoietic cell lines and low grade lymphomas; Gabius S et al.; Important biological functions and cellular recognition phenomena are supposedly governed by specific sugar-protein interactions . Human hematopoietic cell lines offer an excellent model for the study of the expression of endogenous receptors for the carbohydrate part of glycoconjugates with respect to cell lineage and modulation by differentiation . Initially, a panel of fluorescent (neo)glycoproteins was successfully employed to demonstrate cytologically the actual presence of such receptors on different cell lines: the B lymphoblast line, Daudi; the T cell lymphoblastic leukemia line, P12; the multipotent leukemic line, K562 and the promyelocytic line, HL060 . Biochemical analyses were performed using affinity chromatography on supports with immobilized lactose and asialofetuin (simple or complex beta-galactosides), melibiose (alpha-galactoside), fucose, N-acetyl-D-galactosamine, maltose (alpha-glucoside), the mannose-rich yeast glycoprotein, mannan, glycopeptides containing sialic acid residues and heparin . Subsequently, sodium dodecyl sulfate-polyacrylamide slab gel electrophoresis was used to detect cell lineage-dependent changes in theses parameters . Differentiation-dependent changes in the expression of receptors with specificity to galactose, N-acetylgalactosamine, maltose and heparin were similarly uncovered upon dimethyl sulfoxide-induced differentiation of HL60 cells . Differences in this type of cellular characteristic were also apparent for lymphoma cells from patients with various histological subtypes of lowgrade lymphomas . This initial description of lineage- and differentiation-dependent differences in various human hematopoietic cell lines and in cells from patients with lowgrade lymphomas suggests that advances in the knowledge of the composition of endogenous sugar receptors (lectins) may aid in understanding aspects of the biological behavior of hematopoietic cells and their related malignancies via participation of sugar-protein (lectin) interactions.

Am J Clin Nutr, 1989 Aug, 50(2), 346 - 52
Qinghaosu, dietary vitamin E, selenium, and cod-liver oil: effect on the susceptibility of mice to the malarial parasite Plasmodium yoelii; Levander OA et al.; Young female mice were fed torula-yeast-based diets deficient in vitamin E or selenium or supplemented with cod-liver oil to determine the effect of host antioxidant status on the therapeutic efficacy of the Chinese traditional antimalarial drug qinghaosu (QHS), a sesquiterpene endoperoxide . Vitamin E deficiency enhanced the antimalarial action of QHS against Plasmodium yoelii, both in terms of decreased parasitemia and improved survival but Se deficiency did not . A vitamin E-deficient diet containing 5% cod-liver oil had such strong antimalarial activity in itself that no additional therapeutic benefit of QHS could be demonstrated . Hematocrit values in parasitized mice treated with QHS or fed the cod-liver-oil-supplemented, vitamin E-deficient diet were normal . Nutritional manipulation of host antioxidant status may provide a promising prophylactic and/or therapeutic tool for the control of malaria.

Res Commun Chem Pathol Pharmacol, 1989 Aug, 65(2), 249 - 52
Effect of selenium supplementation after acute myocardial infarction; Korpela H et al.; The effect of selenium supplementation was evaluated in 81 patients with acute myocardial infarction in a double-blind, placebo-controlled study . Patients were randomised into two treatment groups receiving either selenium-rich yeast (100 micrograms/day) or placebo in addition to conventional drug therapy for a 6-month period . During treatment the mean serum selenium concentration increased from 82 micrograms/l to 122 micrograms/l (p less than 0.001) in the selenium supplemented group and remained unaltered in the placebo group (83 micrograms/l) . During the 6-month follow-up period there were 4 cardiac deaths in the placebo group whereas no patients in the selenium group died during the follow-up period . Furthermore, there were 2 nonfatal reinfarctions in the placebo group and one nonfatal reinfarction in the selenium group . The results encourage further studies to evaluate the efficacy of antioxidants in the prevention and therapy of acute myocardial infarction.

FASEB J, 1989 Aug, 3(10), 2151 - 63
Structural and functional properties of ras proteins; Santos E et al.; The ras proteins belong to a family of related polypeptides that are present in all eukaryotic organisms from yeast to human . Their extraordinary evolutionary conservation suggests that they have essential cellular functions, although their exact role remains unknown . Mutations in specific amino acids and overexpression of normal proteins have been linked to altered proliferation and/or differentiation and, particularly, to neoplastic processes . Mature ras proteins are located on the inner side of the plasma membrane, and their biochemical properties include binding and exchange of guanine nucleotides and GTPase activity . The favored hypothesis for ras function is that these proteins exist in an equilibrium between an inactive conformation (p21.GDP) and an active conformation (p21 . GTP) in which they are able to interact with their as yet unknown cellular target or targets . Similarities in cellular location, structure, and biochemistry with other known regulatory (G) proteins suggest that they play a role in transduction of signals from the cell surface . The elucidation of the crystal structure of normal and transforming ras proteins and the identification of cellular proteins that interact directly with them (GAP, CDC25) or suppress some of their biological effects (Krev-1) have opened new avenues in the search for their elusive cellular targets and in the elucidation of the functional role of ras gene products.

J Dent Que, 1989 Aug, 26, 425 - 8
{Vaccination as a means of prevention}; Montplaisir S et al.; While recognizing the fact that the use of strict hygienic techniques, disposable materials and quality control are essential for sterilizing instruments, vaccination still remains the most efficient way to combat infection . In this article, the authors discuss the principle problems associated with the use of vaccines and especially as they relate to the dental office . Hepatitis B is certainly of interest to all dentists and their personnel, especially since its primary mode of transmission is by contact of blood with the skin and the mucosa . The two forms of vaccines presently available, are derived from the human plasma of carriers or from yeast and cause a genetic reaction which produces hepatitis B virus antigens . These vaccines protect at least over 90% of all healthy individuals and do not generate any secondary infection or unfavorable reactions . Faced with the reality of hepatitis B, it is very wise to remember that it is BETTER TO BE VACCINATED THAN TO HAVE TO LIVE WITH THE EFFECTS OF HEPATITIS.

Proc Natl Acad Sci U S A, 1989 Aug, 86(15), 5698 - 702
Association of charge clusters with functional domains of cellular transcription factors; Brendel V et al.; Using rigorous statistical methods, we have identified and evaluated unusual properties of the distribution of charged residues within the sequences of eukaryotic cellular transcription factors . Virtually all transcription factors, including GAL4, c-Jun, C/EBP, CREB, Oct-1, Oct-2, Sp1, Egr-1, CTF-1, steroid and thyroid hormone receptors, and others, carry one or more highly significant charge clusters . For the most part these clusters (conserved within families of homologous proteins) are of positive net charge but contain also substantial numbers of acidic residues . Predominantly basic charge clusters are often, but not exclusively, associated with DNA-binding domains, and vice versa . Negative charge clusters of note occur only in the yeast protein PHO4 and in the proteins encoded at the Drosophila loci zeste (zeta) and knrl . This dearth of statistically significant negative charge clusters raises questions with respect to the generality of acidic activation domains . A number of sequences (Oct-1, Oct-2, zeste, Dhr23, E75, and knrl) contain multiple charge clusters together with one or more significantly long uncharged regions . The occurrence of multiple charge clusters is a rare phenomenon (found in less than 3% of all proteins, mainly in Drosophila developmental control proteins and in transactivators of eukaryotic DNA viruses) . Most of the proteins with zinc-binding "fingers" carry a mixed charge cluster centered at the zinc-finger motif preceded by a long uncharged stretch, suggestive of a modular structure for these proteins.

EMBO J, 1989 Aug, 8(8), 2275 - 82
Cyclin is a component of the sea urchin egg M-phase specific histone H1 kinase; Meijer L et al.; A so-called 'growth-associated' or 'M-phase specific' histone H1 kinase (H1K) has been described in a wide variety of eukaryotic cell types; p34cdc2 has previously been shown to be a catalytic subunit of this protein kinase . In fertilized sea urchin eggs the activity of H1K oscillates during the cell division cycle and there is a striking temporal correlation between H1K activation and the accumulation of a phosphorylated form of cyclin . H1K activity declines in parallel with proteolytic cyclin destruction of the end of the first cell cycle . By virtue of the high affinity of the fission yeast p13suc1 for the p34cdc2 protein, H1K strongly binds to p13-Sepharose beads . Cyclin, p34cdc2 and H1K co-purify on this affinity reagent as well as through several conventional chromatographic procedures . Anticyclin antibodies immunoprecipitate the M-phase specific H1K in crude extracts or in purified fractions . Sea urchin eggs appear to contain much less cyclin than p34cdc2, suggesting that p34cdc2 may interact with other proteins . These results demonstrate that cyclin and p34cdc2 are major components of the M-phase specific H1K.

Biokhimiia, 1989 Aug, 54(8), 1294 - 9
{Molecular characteristics of beta-galactosidase secreted by Penicillium canescens}; Nikolaev IV et al.; Extracellular beta-galactosidase from P . canescens culture medium was purified by ion-exchange chromatography on DEAE and CM-Sepharose CL-6B and gel filtration . The enzyme active form was shown to be a monomer with a molecular weight of about 120 kDa; the isoelectric point is 6.7 and the sedimentation coefficient is 6.5 . In terms of physico-chemical and catalytic properties, the purified enzyme is similar to beta-galactosidases of other fungi of genus Penicillium . The amino acid composition and the NH2-terminal sequence of 24 residues non-homologous to the corresponding sequences of bacterial and yeast beta-galactosidases were determined.

Clin Immunol Immunopathol, 1989 Aug, 52(2), 271 - 8
Monocyte function in systemic lupus erythematosus; Boswell J et al.; Systemic lupus erythematosus (SLE) is characterized by multiple defects affecting B and T lymphocyte cell function . In view of the influence that monocytes may have on these functions, we have examined monocyte function in patients with SLE and normals . Monocytes were examined as to number, their ability to modulate IgG synthesis, phagocytose yeast particles, and acid phosphatase activity . Patients had more monocytes and mononuclear cells made more IgG, but these factors appear to be independent of each other . Monocytes from lupus patients are less phagocytic but show similar levels of acid phosphatase activity to normal controls . Normal, but not lupus monocytes, show enhanced phagocytosis following treatment with LPS . The effect of monocyte stimulation on acid phosphatase activity is the same in lupus and normal monocytes . Taken together, these observations suggest that the increased numbers of monocytes in lupus patients and some subtle functional differences in these cells may be important in the progression of this disease.

J Bioenerg Biomembr, 1989 Aug, 21(4), 461 - 70
Hexokinase-binding properties of the mitochondrial VDAC protein: inhibition by DCCD and location of putative DCCD-binding sites; Nakashima RA; The outer mitochondrial membrane receptor for hexokinase binding has been identified as the VDAC protein, also known as mitochondrial porin . The ability of the receptor to bind hexokinase is inhibited by pretreatment with dicyclohexylcarbodiimide (DCCD) . At low concentrations, DCCD inhibits hexokinase binding by covalently labeling the VDAC protein, with no apparent effect on VDAC channel-forming activity . The stoichiometry of {14C}-DCCD labeling is consistent with one to two high-affinity DCCD-binding sites per VDAC monomer . A comparison between the sequence of yeast VDAC and a conserved sequence found at DCCD-binding sites of several membrane proteins showed two sites where the yeast VDAC amino acid sequence appears to be very similar to the conserved DCCD-binding sequence . Both of these sites are located near the C-terminal end of yeast VDAC (residues 257-265 and 275-283) . These results are consistent with a model in which the C-terminal end of VDAC is involved in binding to the N-terminal end of hexokinase.

Biochimie, 1989 Aug, 71(8), 963 - 8
Ionic mitochondrial channels: characteristics and possible role in protein translocation; Henry JP et al.; Most of the mitochondrial proteins are synthesized in the cytoplasm as precursors which are then translocated into the organelle . These precursors have a NH2-terminal extension which functions as a mitochondrial targeting signal . The import process through mitochondrial membranes is voltage-dependent; its mechanism is still unknown . Translocation has been proposed to occur through specific channels, thus, indicating the interest of the study of mitochondrial ionic channels . Two anion channels with different electrical characteristics have been described in the outer and the inner membranes . Using the technique of "Tip-Dip", we have shown the existence of a cation channel of large conductance in mitochondria . The characteristics of this channel differ from that of the other mitochondrial anion channels . A positively charged 13-residue synthetic peptide, with the sequence of the amino terminal extremity of the nuclear-coded subunit IV of yeast cytochrome C oxidase, induces a blockade of the cationic channel . From the characteristics of the blockade, it is likely that the channel could be permeable to the peptide . The specificity of this effect suggests that this channel might be involved in protein translocation.

Exp Cell Res, 1989 Aug, 183(2), 361 - 75
M-phase-specific protein kinase from mitotic sea urchin eggs: cyclic activation depends on protein synthesis and phosphorylation but does not require DNA or RNA synthesis; Arion D et al.; Histone H1 kinase (H1K) undergoes a transient activation at each early M phase of both meiotic and mitotic cell cycles . The mechanisms underlying the transient activation of this protein kinase were investigated in mitotic sea urchin eggs . Translocation of active H1K from particulate to soluble fraction does not seem to be responsible for this activation . H1K activation cannot be accounted for by the transient disappearance of a putative H1K inhibitor present in soluble fractions of homogenates . Aphidicolin, an inhibitor of DNA synthesis, and actinomycin D, an inhibitor of RNA synthesis, do not impede the transient appearance of H1K activity . H1K activation therefore does not require DNA or RNA synthesis . Fertilization triggers a rise in intracellular pH responsible for the increase of protein synthesis . H1K activation is highly dependent on the intracellular pH . Ammonia triggers an increase of intracellular pH and stimulates protein synthesis and H1K activation . Acetate lowers the intracellular pH, decreases protein synthesis, and blocks H1K activation . Protein synthesis is an absolute requirement for H1K activation as demonstrated by their identical sensitivities to emetine concentration and to time of emetine addition . About 60 min after fertilization, H1K activation and cleavage become independent of protein synthesis . The concentration of p34, a homolog of the yeast cdc2 gene product which has been recently shown to be a subunit of H1K, does not vary during the cell cycle and remains constant in emetine-treated cells . H1K activation thus requires the synthesis of either a p34 postranslational modifying enzyme or another subunit . Finally, phosphatase inhibitors and ATP slow down in the in vitro inactivation rate of H1K . These results suggest that a subunit or an activator of H1K is stored as an mRNA in the egg before mitosis and that full activation of H1K requires a phosphorylation.

Cell, 1989 Jul 28, 58(2), 361 - 72
Regulation of p34cdc2 protein kinase during mitosis; Moreno S et al.; The cell-cycle timing of mitosis in fission yeast is determined by the cdc25+ gene product activating the p34cdc2 protein kinase leading to mitotic initiation . Protein kinase activity remains high in metaphase and then declines during anaphase . Activation of the protein kinase also requires the cyclin homolog p56cdc13, which also functions post activation at a later stage of mitosis . The continuing function of p56cdc13 during mitosis is consistent with its high level until the metaphase/anaphase transition . At anaphase the p56cdc13 level falls dramatically just before the decline in p34cdc2 protein kinase activity . The behavior of p56cdc13 is similar to that observed for cyclins in oocytes . p13suc1 interacts closely with p34cdc2; it is required during the process of mitosis and may play a role in the inactivation of the p34cdc2 protein kinase . Therefore, the cdc25+, cdc13+, and suc1+ gene products are important for regulating p34cdc2 protein kinase activity during entry into, progress through, and exit from mitosis.

Gene, 1989 Jul 15, 79(2), 207 - 17
Structure and expression of a gene encoding heat-shock protein Hsp70 from the Oomycete fungus Bremia lactucae; Judelson HS et al.; A gene encoding a protein homologous to a 70-kDa heat-shock protein (Hsp70) was isolated from Bremia lactucae and its structure and pattern of expression were determined . This is the first report on the structure of a protein-coding gene from an Oomycete fungus . The cloned gene is a member of a small multigene family . The level of hsp70 mRNA in germlings increases from a low constitutive level in response to heat or cold treatment . A high level of the mRNA is also detected in spores . The hsp70 gene is expressed as a primary transcript of 2241 nucleotides (nt) and contains a continuous open reading frame of 2025 nt . Near the C terminus of the coding sequence is an unusual region that contains repeated enhancer-like sequences . This insert has not been described in other hsp70 genes and is not an intron . Upstream from the 5' terminus of the mRNA are multiple CCAAT motifs, a sequence similar to a consensus heat-shock regulatory element, and an A + T-rich putative 'TATA' box . A canonical polyadenylation recognition sequence is present downstream from the coding sequence . The deduced amino acid sequence is equally similar to yeast and maize Hsp70, providing further evidence of the dissimilarity between Oomycetes and true fungi . The cloning of this gene is part of our strategy to develop a transformation system for B . lactucae.

J Biol Chem, 1989 Jul 15, 264(20), 11928 - 33
Synthesis and properties of 5-azido-UDP-glucose . Development of photoaffinity probes for nucleotide diphosphate sugar binding sites; Drake RR Jr et al.; A new active site directed photoaffinity probe, which is a model compound for studying nucleotide diphosphate sugar binding proteins, has been synthesized by coupling 5-azido-UTP and {32P}Glc-1-P using yeast UDP-glucose pyrophosphorylase to produce {beta-32P}5-azidouridine 5'-diphosphoglucose (5N3UDP-Glc) . This probe has photochemical properties similar to that of 5-azidoUTP (Evans, R . K., and Haley, B . E . (1987) Biochemistry 26, 269-276) . The efficacy of 5N3UDP-Glc as an active site directed probe was demonstrated using yeast UDP-Glc pyrophosphorylase . Saturation effects of photoinsertion were observed with an apparent Kd of 51 microM and the natural substrate, UDP-Glc, prevented photoinsertion of {beta-32P}5N3UDP-Glc with an apparent Kd of 87 microM . Prevention of photoinsertion was also seen with UTP and pyrophosphate with apparent Kd values less than 200 microM . UMP, UDP, ATP, and GTP were much less effective competitors . Selective photoinsertion was observed with several partially purified enzymes including UDP-Glc dehydrogenase, UDP-Gal-4-epimerase, Gal-1-P uridyltransferase, and phosphorylase a . The absence of nonselective photoinsertion into bulk proteins was demonstrated with crude homogenates of rabbit liver as well as with several UDP-Glc binding proteins . Of the six purified enzymes tested, only phosphoglucomutase has been shown to incorporate radiolabel from the photoprobe in the absence of UV irradiation . These results and a discussion of the utility of 5N3UDP-Glc for detecting UDP-Glc binding proteins and isolating active site peptides are presented.

Cell, 1989 Jul 14, 58(1), 69 - 76
Evolution of plant mitochondrial genomes via substoichiometric intermediates; Small I et al.; Comparison of the modern fertile maize mitochondrial genome (N) with an ancestral maize mitochondrial genome (RU) reveals a 12 kb duplication (containing the atpA gene) in the modern genome that is absent from the ancestor . Cloning, mapping, and sequencing of the relevant portions of the ancestral genome shows that this duplication probably arose via a three-stage recombination process involving substoichiometric intermediates . Comparison with analogous observations on yeast mitochondrial genomes suggests that this three-stage model of genome reorganization can be generally applied to plant mitochondrial genomes to explain both deletions and the creation of novel repeats, common features of plant mitochondrial genome evolution.

Cell, 1989 Jul 14, 58(1), 37 - 45
Site selection by Xenopus laevis RNAase P; Carrara G et al.; Investigation of the mechanism of cleavage site selection by Xenopus RNAase P reveals that the acceptor stem, a 7 bp helix common to all tRNA precursors, is required for cleavage . We propose that Xenopus RNAase P recognizes conserved features of the mature tRNA and that the cleavage site is selected by measuring the length of the acceptor stem . In support of this, we demonstrate that insertion of 2 bp in the acceptor stem of yeast pre-tRNA(3Leu) relocates the cleavage site 2 bases 3' to the original one . In addition, insertion of 1 bp in the acceptor stem of the end-matured yeast pre-tRNA(Phe) generates an RNAase P cleavage site: the enzyme produces a mature tRNA with the characteristic 7 bp stem and releases one 5' flanking nucleotide . Since it has previously been shown that cleavage sites of the splicing endonuclease are determined by the length of the anticodon stem, RNAase P and the splicing endonuclease apparently use different stems to determine their cutting sites.

Cell, 1989 Jul 14, 58(1), 85 - 93
Transcriptional activation by the v-myb oncogene and its cellular progenitor, c-myb; Weston K et al.; The v-myb oncogene, like its cellular progenitor c-myb, encodes a short-lived nuclear protein involved in processes affecting growth and differentiation in a number of cell types . Fusion proteins, in which v-myb sequences are linked to the DNA binding domain of the yeast transcriptional activator GAL4, can activate transcription from a reporter gene linked in cis to a GAL4 binding site . The domain of v-myb responsible for transcriptional activation is located between residues 204 and 254, and is both necessary and sufficient for activation . Intact v-myb and c-myb proteins can also activate transcription, via a myb binding site linked in cis to a reporter gene . A v-myb protein bearing a deletion in the activator domain is no longer capable of stimulating transcription.

Biochemistry, 1989 Jul 11, 28(14), 5794 - 801
Structure of an unmodified tRNA molecule; Hall KB et al.; We have used NMR to study the structure of the yeast tRNA(Phe) sequence which was synthesized by using T7 RNA polymerase . Many resonances in the imino 1H- spectrum of the transcript have been assigned, including those of several tertiary interactions . When the Mg2+ concentration is high, the transcript appears to fold normally, and the spectral features of the transcript resemble those of tRNA(Phe) . The transcript has been shown to be aminoacylated with kinetics similar to the modified tRNA(Phe) {Sampson, J . R., & Uhlenbeck, O . C . (1988) Proc . Natl . Acad . Sci . U.S.A . 85, 1033-1037}, suggesting that the structure of the two molecules must be similar . In the absence of Mg2+ or at {tRNA}:{Mg2+} ratios less than 0.2, the transcript does not adopt the native structure, as shown by both chemical shifts and NOE patterns . In these low Mg2+ conditions, a second GU base pair is found, suggesting a structural rearrangement of the transcript . NMR data indicate that the structure of a mutant having G20 changed to U20 is nearly identical with that of the normal sequence, suggesting that the low aminoacylation activity of this variant is not due to a substantially different conformation.

Biochemistry, 1989 Jul 11, 28(14), 6035 - 41
A novel, arsenite-sensitive E2 of the ubiquitin pathway: purification and properties; Klemperer NS et al.; In the multienzyme ubiquitin-dependent proteolytic pathway, conjugation of ubiquitin to target proteins serves as a signal for protein degradation . Rabbit reticulocytes possess a family of proteins, known as E2's, that form labile ubiquitin adducts by undergoing transthiolation with the ubiquitin thiol ester form of ubiquitin activating enzyme (E1) . Only one E2 appears to function in ubiquitin-dependent protein degradation . The others have been postulated to function in regulatory ubiquitin conjugation . We have purified and characterized a previously undescribed E2 from rabbit reticulocytes . E2(230K) is an apparent monomer with a molecular mass of 230 kDa . The enzyme forms a labile ubiquitin adduct in the presence of E1, ubiquitin, and MgATP and catalyzes conjugation of ubiquitin to protein substrates . Exogenous protein substrates included yeast cytochrome c(Km = 125 mu M; kcat approximately 0.37 min-1) and histone H3 (Km less than 1.3 mu M; kcat approximately 0.18 min-1) as well as lysozyme, alpha-lactalbumin, and alpha-casein . E2(230K) did not efficiently reconstitute Ub-dependent degradation of substrates that it conjugated, either in the absence or in the presence of the ubiquitin-protein ligase that is involved in degradation . E2(230K) may thus be an enzyme that functions in regulatory Ub conjugation . Relative to other E2's, which are very iodoacetamide sensitive, E2(230K) was more slowly inactivated by iodoacetamide (k(obs) = 0.037 min-1 at 1.5 mM iodoacetamide; pH 7.0, 37 degrees C) . E2(230K) was also unique among E2's in being subject to inactivation by inorganic arsenite (k(i)max = 0.12 min-1; K(0.5) = 3.3 mM; pH 7.0, 37 degrees C) . Arsenite is considered to be a reagent specific for vicinal sulfhydryl sites in proteins, and inhibition is usually rapidly reversed upon addition of competitive dithiol compounds . Inactivation of E2(230K) by arsenite was not reversed within 10 min after addition of dithiothreitol at a concentration that blocked inactivation if it was premixed with arsenite; inactivation is therefore irreversible or very slowly reversible . We postulate that a conformation change of E2(230K) may be rate-limiting for interaction of enzyme thiol groups with arsenite.

Biochim Biophys Acta, 1989 Jul 7, 1008(2), 243 - 6
Transformation of Neurospora crassa by an integrative transforming plasmid is not enhanced by ribosomal DNA sequences; Russell PJ et al.; Two integrative transforming plasmids of Neurospora crassa that differed only by the presence of almost all of a ribosomal DNA repeat unit on one plasmid were constructed . The plasmids were used to test the target concentration hypothesis which states that the transformation frequency is proportional to the number of genomic copies of a homologous sequence located on the transforming plasmid . Since there are approx . 200 copies of the rDNA sequences in the genome, the target concentration hypothesis would have been proved if the transformation frequency was 200-fold higher for the rDNA-containing plasmid compared with the plasmid without rDNA . The results indicated no difference in the transformation for the two plasmids, thereby providing no support for the hypothesis . The target concentration hypothesis has been proved for yeast, and thus mechanisms different from that responsible for integrative transformation in yeast must operate in N . crassa, perhaps including non-homologous recombination events.

Vopr Med Khim, 1989 Jul-Aug, 35(4), 132 - 4
{Determination of glutathione peroxidase activity}; Model' MA; A simplified procedure is developed for estimation of glutathione peroxidase activity involving glutathione reductase in the conjugated reaction . The procedure developed enabled to use commonly employed spectrophotometers and to decrease distinctly the glutathione reductase consumption . Isolation of glutathione reductase from yeast is described.

Mycopathologia, 1989 Jul, 107(1), 51 - 5
Ketoconazole resistance in Torulopsis glabrata; Nobre G et al.; The majority of clinical isolates of T . glabrata has been shown highly sensitive to ketoconazole, when tested with an agar dilution method, and resistant, when using a broth dilution method . The fact was accounted for the high degree of mutation associated with the haploid state present in this species . Experimental T . glabrata infection in immunocompetent and immunocompromised mice appears not respond to the oral administration of the drug . The results agree with that of broth dilution tests and not with those produced by the agar methods . It seems that agar dilution sensitivity tests are inadequate for yeast species with an high rate of mutation, as T . glabrata.

Nippon Shokakibyo Gakkai Zasshi, 1989 Jul, 86(7), 1519 - 24
{The effects of biotin on the metabolism of ammonia and amino acids in urease-induced hyperammonemic rats}; Nagamine T et al.; The effects of oral and intraperitoneal administration of biotin in urease-induced hyperammonemic rats, as well as the influence of biotin deficiency, have been studied . Biotin deficiency was produced by feeding standard diet MF (Oriental Yeast Co.) supplemented with dry egg-white (egg-white group) . Egg-white + biotin group had free access to 0.0014% of biotin solution at all time . Following an intraperitoneal injection of urease, 25 U/kg (B.W.), plasma ammonia levels in egg-white + biotin group were lower than in egg-white group, especially there was significance (p less than 0.05) at 8 hours after the urease injection . Similarly, plasma ammonia levels in biotin-injected rats, in which 1 mg of biotin had been injected intraperitoneally prior to the experiment, were significantly low compared with saline-injected controls at 4 and 6 hours after urease administration . Results of plasma amino acid analysis, 9 hours after the urease injection indicated that Fischer's molar ratio (Leu + Ileu + Val/Tyr + Phe) was significantly higher in the biotin-injected rats than the saline-injected control . It suggests that biotin might decrease blood ammonia by facilitating the detoxification mechanism as follow: L-glutamate + NH3----L-glutamine.

J Biolumin Chemilumin, 1989 Jul, 4(1), 159 - 63
Enhanced chemiluminescence ELISA for the detection of antibody to hepatitis B virus surface antigen; Ireland D et al.; A chemiluminescent enzyme linked immunosorbent assay (ELISA) for the detection of antibody to hepatitis B virus surface antigen (anti-HBs) in human serum has been developed . Polystyrene microtitre plates were coated with recombinant, yeast-derived hepatitis B surface antigen (rec-HBsAg) . Patient serum samples and appropriate controls were added to the rec-HBsAg-coated wells and incubated to bind anti-HBs . The wells were then washed and a fluorescein isothiocyanate (FITC) conjugate of a human plasma-derived hepatitis B surface antigen (HBsAg) was added . Following incubation and further washing, the bound FITC-labelled HBsAg was detected after addition of a horseradish peroxidase (HRP) conjugate of a monoclonal anti-FITC antibody and assaying for the enzyme . The activity of the HRP was measured using luminol and hydrogen peroxide as substrates and iodophenol as a chemiluminescence enhancer . The luminescence was recorded using a camera luminometer . Preliminary tests have shown the assay to be suitable for the detection of antibody in sera from both vaccinees and also from individuals with a past hepatitis B virus infection . The use of the FITC-anti-FITC system together with the measurement of a chemiluminescence signal makes possible the completion of this assay in a few hours . The assay has been shown to be both specific and sensitive and provides a permanent photographic record.

Res Commun Chem Pathol Pharmacol, 1989 Jul, 65(1), 35 - 56
Research on heterocyclic compounds . XXV . Phenyl derivatives of fused imidazole systems: antiinflammatory activity; Abignente E et al.; A group of seven acidic phenyl derivatives of some fused imidazole systems were subjected to a series of tests in vivo in order to evaluate their pharmacological activity . Antiinflammatory activity was studied by means of the carrageenin-induced rat paw edema . Hot plate test and writhing induced by acetic acid in mice were used to evaluate analgesic activity . Yeast-induced hyperthermia in rats was employed to study antipyretic action . Irritative and ulcerogenic action on the rat gastric mucosa was examined at higher doses . The antiaggregating activity and the inhibition of platelet malondialdehyde production were studied in vitro . All experimental results are discussed from the point of view of structure-activity relationships and mode of action.

S Afr Med J, 1989 Jul 1, 76(1), 13 - 6
Blastomyces dermatitidis infections in the RSA; Frean JA et al.; Twenty cases of blastomycosis have been confirmed in the RSA, 9 of which are presented for the first time . Patients came from all four provinces and the mean age was 40 years . Six cases were diagnosed between 1985 and 1987 . Differences between strains of Blastomyces dermatitidis isolated in the RSA and in North America include morphological and cultural characteristics, mycelial-yeast conversion, antigenic structure, and compatibility in cross-mating experiments . The diagnosis of this disease can be made by direct examination of unstained specimens, by histological examination or by culture of the organism . Culture should be attempted in all cases for confirmation of microscopic findings.

Res Microbiol, 1989 Jul-Aug, 140(6), 373 - 8
Differentiation of Candida guilliermondii varieties by lectin-like substances from marine algae; Fabregas J et al.; Lectin-like substances of 54 marine algae, corresponding to 30 red algae and 24 brown algae, were studied to estimate the agglutination activity against Candida guilliermondii . Extracts of Dictyopteris membranacea, Bifurcaria bifurcata, Halidrys siliquosa, Ascophyllum nodosum, Fucus serratus and F . spiralis algae showed selective agglutinating activities for C . guilliermondii "sensu stricto" and some of its varieties . According to their response to agglutination (positive or negative), certain algal extracts enabled the differentiation of varieties of C . guilliermondii independently of the titre values . Three extracts of brown algae were sufficient to identify 4 varieties of C . guilliermondii . The results suggest that lectin-like substances from marine algae may be valuable reagents as assistant tools for the identification of yeast strains.

Vestn Khir Im I I Grek, 1989 Jul, 143(7), 39 - 43
{Preventive use of immunostimulants in patients with infectious endocarditis}; Kostiuchenko AL et al.; Leakadine++, levamisole++, proper-myl were used in the postoperative period in 36 patients operated upon for infectious endocarditis with regard for the character of the immunopathology found . The directed immunostimulation results in resolution of immunodeficiency and normalization of the immune status of the patients.

Mol Biol (Mosk), 1989 Jul-Aug, 23(4), 1171 - 6
{Characteristics of DNA isolated from a complex form of DNA-polymerase alpha from the rat liver}; Timchenko NA et al.; A complex from of DNA polymerase alpha was isolated from the nuclear membrane of hepatocytes . DNA fragments were shown to be among components of the complex under study . In this paper we present evidence that DNA from the alpha-polymerase complex from quiescent hepatocytes (DNA-G) differs in its nucleotide composition from its counterpart (DNA-S) isolated from hepatocytes synthesizing DNA . As judged by dot hybridization, DNA-G0 does not contain nucleotide sequences which are complementary to ribosomal or messenger RNA, whereas the abovementioned sequences are present in DNA-S . At the same time DNA-G0 is found to contain sequences which are homologous to both SV40 DNA and yeast TRPI-ARS1 DNA . The difference in nucleotide sequences between DNA-G0 and DNA-S indicates that in the process of replication DNA is being stretched across the multienzyme complex located on the nuclear membrane.

Prostaglandins, 1989 Jul, 38(1), 79 - 89
Effects of inadequate vitamin E and/or selenium nutrition on the release of arachidonic acid metabolites in rat alveolar macrophages; Eskew ML et al.; Effects of vitamin E and/or selenium (Se) deficiency on the secretion of arachidonic acid metabolites by zymosan-stimulated pulmonary alveolar macrophages (AM) were examined using cells from male Long-Evans hooded rats fed torula-yeast based diets with or without the supplementation of vitamin E (150 IU/kg) or Se (0.5 mg/kg) . Alveolar macrophages obtained by lavage were purified by adherence and cultured for 4 h in Hank's balanced salt solution containing bovine serum albumin (0.1%) and zymosan (300 micrograms/ml) . The arachidonic acid metabolites present in the culture supernatant were measured by radioimmunoassay . Altered vitamin E and Se nutrition had no effect on the number of cells or cell types recovered from the pulmonary airways . Alveolar macrophages derived from animals fed on diets deficient in vitamin E or Se or both nutrients secreted higher levels of prostaglandin E2 and thromboxane B2 . Levels of both 5-hydroxyeicosatetraenoic acid and leukotriene B4 were significantly increased only in the group fed the diet adequate in Se but deficient in vitamin E . Our data suggest that vitamin E and Se might play an important role to control the levels of several physiologically and pathologically important arachidonic acid metabolites.

Proc Natl Acad Sci U S A, 1989 Jul, 86(14), 5325 - 9
Molecular cloning of rat homologues of the Drosophila melanogaster dunce cAMP phosphodiesterase: evidence for a family of genes; Swinnen JV et al.; To study the structure and function of cyclic nucleotide phosphodiesterases (PDEs) involved in mammalian gametogenesis, a rat testis cDNA library was screened at low stringency with a cDNA clone coding for the Drosophila melanogaster dunce-encoded PDE as a probe . This screening resulted in the isolation of two groups of cDNA clones, differing in their nucleotide sequences (ratPDE1 and ratPDE2) . In the rat testis, RNA transcripts corresponding to both groups of clones were expressed predominantly in germ cells . Additional screenings of a Sertoli cell cDNA library with a ratPDE2 clone as a probe led to the isolation of two more groups of clones (rat-PDE3 and ratPDE4) . Unlike ratPDE1 and ratPDE2, these clones hybridized to transcripts present predominantly in the Sertoli cell . In the middle of the coding region, all four groups of clones were homologous to each other . The deduced amino acid sequences of part of this region were also homologous to the D . melanogaster dunce PDE and to PDEs from bovine and yeast . These data indicate that a family of genes homologous to the D . melanogaster dunce-encoded PDE is present in the rat and that these genes are differentially expressed in somatic and germ cells of the seminiferous tubule . These findings provide a molecular basis for the observed heterogeneity of cAMP PDEs.

Allergol Immunopathol (Madr), 1989 Jul-Aug, 17(4), 205 - 7
Polymorphonuclear functions in patients with mixed connective tissue disease; Bodolay E et al.; Polymorphonuclear leukocytes (PMNs) from 29 patients with mixed connective tissue disease (MCTD) were studied "in vitro" for their phagocytic and chemotactic function as well as for granulocyte alkaline phosphatase (GAP) activity . Fc-receptor expression detected by EA-rosette formation was comparable to the control . Yeast-phagocytosis, C3b-receptor mediated phagocytosis and chemotaxis of PMNs, however, significantly decreased in MCTD . At the same time, photometric measure of alkaline phosphatase activity indicated a nearly two fold increase in PMNs from patients with MCTD . Although no correlation was found between PMN functions and the activity of the disease, PMN disorders may play a role in pathogenesis of these connective tissue disorders.

Vet Med (Praha), 1989 Jul, 34(7), 421 - 30
{Detection of cyclopiazonic acid and its producers in food}; Ostry V et al.; In the course of six months, 60 samples of foods were examined for their contents of cyclopiazonic acid . These samples were subjected to a basal mycological screening aimed at Aspergillus flavus and Penicillium sp . strains . Cyclopiazonic acid contents in samples of Hermelin cheese, peanuts, rice, peeled barley grains, Folican salami, and packaged meat did not exceed the value of 0.5 mg.kg-1 . When using a modification of the method of cyclopiazonic acid isolation described by Dorner et al . (1983), 521 mg of this mycotoxin were isolated from a culture of Penicillium griseofulvum CCM 8006 strain grown in liquid medium containing 2% yeast autolysate and 2.5% sucrose . About 47% of the isolated Aspergillus flavus strains were bitoxicogenic (produced both cyclopiazonic acid and aflatoxin) . Cyclopiazonic acid was produced by 23.5% of the isolated Penicillium sp . strains . No cyclopiazonic acid was produced in vitro by Penicillium nalgoviensis strains from the Czechoslovak collection on sweet wort agar containing peptone from soybean . Penicillium commune F-426 and Penicillium aurantiogriseum F-708 strains are efficient producers of this acid.

J Mol Evol, 1989 Jul, 29(1), 40 - 51
Dinoflagellates in evolution . A molecular phylogenetic analysis of large subunit ribosomal RNA; Lenaers G et al.; The sequence of the large subunit ribosomal RNA (LsuRNA) gene of the dinoflagellate Prorocentrum micans has been determined . The inferred rRNA sequence {3408 nucleotides (nt)} is presented in its most probable secondary structure based on compensatory mutations, energy, and conservation criteria . No introns have been found but a hidden break is present in the second variable domain, 690 nt from the 5' end, as judged by agarose gel electrophoresis and primer extension experiments . Prorocentrum micans LsuRNA length and G+C content are close to those of ciliates and yeast . The conserved portions of the molecule (1900 nt) have been aligned with corresponding sequences from various eukaryotes, including five protista, one metaphyta, and three metazoa . An extensive phylogenetic study was performed, comparing two phenetic methods (neighbor joining on difference matrix, and Fitch and Margoliash on Knuc values matrix) and one cladistic (parsimony) . The three methods led to similar tree topologies, except for the emergence of yeast that groups with ciliates and dinoflagellates when phenetic methods are used, but emerges later in the most parsimonious tree . This discrepancy was checked by statistical analyses on reduced trees (limited to four species) inferred using parsimony and evolutionary parsimony methods . The data support the phenetic tree topologies and a close relationship between dinoflagellates, ciliates, and yeast.

Proc Natl Acad Sci U S A, 1989 Jul, 86(13), 4892 - 6
Multiple alpha subunits of guanine nucleotide-binding proteins in Dictyostelium; Pupillo M et al.; Previous results have shown that chemotaxis and the expression of several classes of genes in Dictyostelium discoideum are regulated through a cell surface cAMP receptor interacting with guanine nucleotide-binding proteins (G proteins) . We now describe cloning and sequencing of cDNAs encoding two G alpha protein subunits from Dictyostelium . The derived amino acid sequences show that they are 45% identical to each other and to G alpha protein subunits from mammals and yeast . Both cDNAs are complementary to multiple mRNAs that are differentially expressed during development . This evidence and analysis of mutants presented elsewhere suggest that they have distinct physiological functions.

Arch Biochem Biophys, 1989 Jul, 272(1), 97 - 102
Phytoalexin synthesis in soybean cells: elicitor induction of reductase involved in biosynthesis of 6'-deoxychalcone; Welle R et al.; Chromatofocusing on Mono P proved to be an efficient purification procedure for the NADPH-dependent reductase from soybean (Glycine max L.) cell cultures which acts together with chalcone synthase in the biosynthesis of 2',4',4-trihydroxychalcone (6'-deoxychalcone) . By isoelectric focusing the pI of reductase was determined to be 6.3 . Addition of pure soybean reductase to cell-free extracts from stimulated cell cultures of parsley and bean (Phaseolus vulgaris) and from young flowers of Dahlia variabilis caused in each case synthesis of 6'-deoxychalcone . When 4-coumaroyl-CoA was replaced by caffeoyl-CoA in the reductase assay, formation of 2',4',3,4-tetrahydrochalcone (butein) was observed . A polyclonal antireductase antiserum was raised in rabbits and proved to be specific in Ouchterlony diffusion experiments, Western blots and immunotitration . The reductase antiserum showed no cross-reactivity with soybean chalcone synthase (CHS) . A biotin/{125I}streptavidin system provided a quantitative Western blot for the reductase . Changes in the activities, amounts of protein, and mRNA activities of reductase and CHS were determined after challenge of soybean cell cultures by elicitor (from Phytophthora megasperma f.sp . glycinea or yeast) . For both enzymes a pronounced and parallel increase in activity and amounts of protein was observed after elicitor addition with a maximum at about 16 h after challenge . Parallel increases in mRNA activities occurred earlier . The results indicate a parallel induction of de novo synthesis of reductase and CHS which coact in synthesis of 6'-deoxychalcone.

Mol Plant Microbe Interact, 1989 Jul-Aug, 2(4), 165 - 8
Expression of reverse transcriptase genes in Fulvia fulva; McHale MT et al.; Antibodies raised against intercellular fluid antigens isolated from diseased tomato leaves have revealed that the fungal pathogen Fulvia fulva expresses genes for a fungal reverse transcriptase (RNA-dependent DNA polymerase) . This enzyme is required for the replication of retroviruses and retroviral-like transposable elements and could provide a mechanism for increasing the mutation rate of fungal pathogens, perhaps explaining their ability to evolve new races rapidly . We report here the DNA sequence of a 225-bp clone from a lambda gt11 genomic library of F . fulva . This clone, designated P5, exhibits a high degree of sequence homology with the reverse transcriptase (pol) gene of the Drosophila melanogaster copia-like retrotransposon 17.6 . Southern blot analysis of genomic DNA of F . fulva showed that P5-related sequences are moderately reiterated with 30-100 copies, some of which exhibit restriction fragment length polymorphism in different races of the pathogen . Western blot analysis of extracts from F . fulva with antibodies raised to purified reverse transcriptase (from human immunodeficiency virus-1) revealed immunoreactive proteins . Reverse transcriptase previously has been detected in a variety of organisms including yeast, insects, protozoa, and mammals, but to our knowledge, this is the first report of its occurrence in filamentous fungi.

Cell, 1989 Jun 30, 57(7), 1167 - 77
All ras proteins are polyisoprenylated but only some are palmitoylated; Hancock JF et al.; The C-terminal CAAX motif of the yeast mating factors is modified by proteolysis to remove the three terminal amino acids (-AAX) leaving a C-terminal cysteine residue that is polyisoprenylated and carboxyl-methylated . Here we show that all ras proteins are polyisoprenylated on their C-terminal cysteine (Cys186) . Mutational analysis shows palmitoylation does not take place on Cys186 as previously thought but on cysteine residues contained in the hypervariable domain of some ras proteins . The major expressed form of c-K-ras (exon 4B) does not have a cysteine residue immediately upstream of Cys186 and is not palmitoylated . Polyisoprenylated but nonpalmitoylated H-ras proteins are biologically active and associate weakly with cell membranes . Palmitoylation increases the avidity of this binding and enhances their transforming activity . Polyisoprenylation is essential for biological activity as inhibiting the biosynthesis of polyisoprenoids abolishes membrane association of p21ras.

Cell, 1989 Jun 30, 57(7), 1259 - 73
The gradient morphogen bicoid is a concentration-dependent transcriptional activator; Struhl G et al.; The bicoid (bcd) protein is expressed in an anteroposterior gradient in early Drosophila embryos and controls the zygotic activation of the segmentation gene hunchback (hb) in a broad but precisely bounded anterior domain . Here we show that the hb gene contains multiple regulatory elements that mediate transcriptional activation in response to bcd protein . Further, we demonstrate that the resulting patterns of expression in vivo depend critically on both the bcd gradient profile and the number and quality of these hb elements . Finally, we show that these same elements mediate bcd-dependent transcriptional activation in yeast and that this interaction requires distinct DNA binding and activating regions in the bcd protein . Our results argue that bcd protein normally binds and activates the hb gene in a concentration-dependent fashion, thereby allowing the gradient of bcd protein to dictate where the hb gene is initially turned on in early embryos . They also suggest that the bcd gradient has the instructive capacity to activate other subordinate control genes by the same mechanism, each in a distinct spatial domain according to its affinity for bcd protein.

J Biol Chem, 1989 Jun 25, 264(18), 10649 - 53
The two Xenopus laevis SRC genes are co-expressed and each produces functional pp60src; Steele RE et al.; The haploid genome of Xenopus laevis contains two src genes, and transcripts from both genes are found in the maternal RNA pool of the oocyte (Steele, R . E . (1985) Nucleic Acids Res . 13, 1747-1761) . We have now isolated cDNA clones which contain complete coding sequences from both src mRNAs . In vitro translation of RNAs transcribed in vitro from these clones produces in each case a protein with an apparent molecular mass of 57 kDa . The in vitro-synthesized proteins show identical protease cleavage patterns . Sequence analysis of the coding regions of the two cDNAs revealed that they both produce 532-amino acid polypeptides which differ from each other at only eight sites . Analysis of silent site changes between the two coding sequences suggests that the two genes began diverging about 25 million years ago . Hybridization with probes specific for each of the two src RNAs indicates that the two genes are co-expressed in embryos and in at least some adult tissues as well as during oogenesis . Finally, expression of each of the cDNA clones in yeast causes the appearance of proteins which are recognized by an antibody which binds to phosphotryosine.

Science, 1989 Jun 23, 244(4911), 1475 - 7
Regulatory role for GTP-binding proteins in endocytosis; Mayorga LS et al.; Guanosine 5'-triphosphate (GTP)-binding proteins have been implicated in the transport of newly synthesized proteins along the secretory pathway of yeast and mammalian cells . Early vesicle fusion events that follow receptor-mediated endocytosis as measured by three in vitro assays were blocked by guanosine 5'-O-(3-thiotriphosphate) and aluminum fluoride . The effect was specific for guanosine nucleotides and depended on the presence of cytosolic factors . Thus, GTP-binding proteins may also have a role in the transport of molecules along the endocytic pathway.

Carbohydr Res, 1989 Jun 15, 189, 289 - 99
Purification of endo-N-acetyl-beta-D-glucosaminidase H by substrate-affinity chromatography; Greber UF et al.; Endo-N-acetyl-beta-D-glucosaminidase H (Endo H) was purified to homogeneity (3000-fold) from a culture filtrate to Streptomyces plicatus . The key step was substrate-affinity chromatography, which afforded a 1000-fold purification and yielded a protease- and exoglycosidase-free preparation of Endo H . Proteins from the crude sample were applied to the substrate-affinity column, consisting of yeast-invertase glycopeptides bound to Sepharose-immobilized concanavalin A . After washing off the unbound proteins, Endo H was quantitatively eluted by methyl alpha-D-mannopyranoside . Various conditions were tested to achieve an optimal binding of Endo H to this substrate-affinity gel . After substrate-affinity chromatography, Endo H was separated from the coeluted gylcopeptide substrate and some protein impurities by gel filtration and hydrophobic chromatography.

Biochemistry, 1989 Jun 13, 28(12), 5226 - 31
Molecular organization of developmentally regulated Dictyostelium discoideum ubiquitin cDNAs; Ohmachi T et al.; Dictyostelium discoideum ubiquitin mRNAs are regulated in a complex fashion during spore germination and multicellular development . Species of mRNA of 1900, 1400, 1100, 840, 580, and 500 nucleotides (nt) are found which are expressed differentially during different stages of development . DNA blot analysis indicates that ubiquitin genes constitute a multigene family of at least six genes . cDNAs representing all the ubiquitin mRNA transcripts were isolated and sequenced . The Dictyostelium mRNAs are organized as tandem repeats of the 76 amino acid ubiquitin unit (228 nt) . We isolated one cDNA containing seven of these tandem repeats, and two different five- and three-repeat cDNAs . In addition, 2 cDNAs containing a single ubiquitin repeat fused at its 3' end to an unrelated 52 and 78 amino acid extension were identified . There is a remarkable similarity in the sequences of the non-ubiquitin extensions among yeast and mammalian counterparts . The extensions are very basic, containing approximately 30% lysine/arginine . Another common feature of these proteins is the presence of a common structural motif containing cysteine residues at conserved positions, suggesting a metal binding domain that matches a consensus sequence of Xenopus transcription factor TFIIIA and other nucleic acid binding proteins . The characterization of ubiquitin cDNAs and genomic sequences in D . discoideum now makes the understanding of its developmental regulation feasible.

J Biol Chem, 1989 Jun 5, 264(16), 9141 - 4
A synthetic peptide-directed antibody as a probe of the phosphorylation site of pyruvate dehydrogenase; Miernyk JA et al.; A synthetic peptide, corresponding to the 14-amino acid tryptic fragment containing phosphorylation sites one and two of bovine mitochondrial pyruvate dehydrogenase, was coupled to Limulus polyphemus hemocyanin and used to produce rabbit polyclonal antibodies . A positive signal was observed when Western blots of bovine, porcine, or yeast mitochondrial pyruvate dehydrogenase complexes were probed with the antibodies but not with blots of bacterial, cellular slime mold, plant mitochondrial, or plant plastid pyruvate dehydrogenase complexes . The antibodies also gave a positive signal when used to probe blots of the bovine kidney branched chain 2-oxo acid dehydrogenase complex . The ATP-dependent phosphorylation/inactivation of rat liver mitochondrial pyruvate dehydrogenase complex could be inhibited by prior incubation with the anti-peptide antibodies.

J Biol Chem, 1989 Jun 5, 264(16), 9103 - 6
Guanylate cyclase, a cell surface receptor; Garbers DL; Guanylate cyclase appears to represent a central member of a diverse family of proteins involved in cell signaling mechanisms including the protein kinases, a low Mr ANP receptor, and possibly adenylate cyclase (based on limited sequence identity with the yeast enzyme) . A membrane form of guanylate cyclase represents a new model for cell surface receptors, although such a model was once envisioned for adenylate cyclase (79) . In original models for adenylate cyclase, hormone was thought to bind with either the enzyme or with an unknown protein to enhance cyclic AMP production (79) . Guanylate cyclase appears to fall into the first adenylate cyclase model where binding of a ligand to an extracellular site on the enzyme transmits a signal to an intracellular catalytic site . The production of cyclic GMP, a second messenger, and of pyrophosphate are then increased . The protein tyrosine kinase family of receptors (80) and possibly another forthcoming family of cell surface receptors containing protein tyrosine phosphatase activity (81-83) contain a single transmembrane domain like guanylate cyclase . Furthermore, the protein tyrosine kinases are activated by ligand binding to the extracellular domain . However, the activation of guanylate cyclase, unlike these cell surface receptors, results in the formation of a low molecular weight second messenger.

Cell, 1989 Jun 2, 57(5), 835 - 45
The genes and transcripts of an antigen gene expression site from T . brucei; Pays E et al.; The AnTat 1.3A antigen gene expression site of T . brucei was cloned from genomic libraries of the 200 kb expressor chromosome . In addition to the antigen gene, it contains seven putative coding regions (ESAGs, for expression site-associated genes), as well as a RIME retroposon . The polypeptide encoded by ESAG 4 shows homology to yeast adenylate cyclase, and possesses structural features of a transmembrane protein . The expression site is transcribed by a pol l-like polymerase in the parasite bloodstream form only, but sequences similar to ESAGs 5, 4, and 2 are also transcribed constitutively elsewhere, by a polymerase sensitive to alpha-amanitin . Ultraviolet irradiation, which seems to block RNA processing, allows the tentative mapping of a transcription promoter about 45 kb upstream of the antigen gene.

Vrach Delo, 1989 Jun, (6), 10 - 2
{Optimization of chemotherapy of stomach cancer}; Chernyi VA et al.; Results are analyzed of the treatment of 520 patients with diffuse gastric cancer using different schemes that included chemotherapy with fluorouracil, intraabdominal polychemotherapy with fluorouracil and adriablastin, immunotherapy with yeast polysaccharides and thymus hormone--taktivin . Chemotherapy supplemented by immunotherapy essentially increases survival . Intraabdominal administration of antitumour drugs in gastric cancer is anatomically more adequate than the intravenous route.

Akush Ginekol (Mosk), 1989 Jun, (6), 20 - 3
{Urogenital chlamydia infection in women with habitual abortion}; Chuchupalov PD et al.; The Chlamydozoan population was identified in 43.6 per cent of 110 females with a history of habitual abortion . It was found that the infection was more frequent in females aged under 30 years, in those who was born preterm, in patients with early (premarital) sexual contacts or those with spontaneously stopped pregnancy during the first trimester, as well as in those females who suffered from cervicitis, yeast colpitis, pyelonephritis or had post-inflammatory changes of placental tissue . Higher incidence of cervicitis, cervical erosion, yeast colpitis, salpingo--oophoritis and pyelonephritis was documented in females infected with Chlamydozoa.

Lipids, 1989 Jun, 24(6), 518 - 25
A novel spectrophotometric assay for lipase activity utilizing cis-parinaric acid; Rogel AM et al.; A new spectrophotometric assay for determining the activity of acylglycerol hydrolases (lipases, E.C . 3.1.1.3) was developed and optimized for yeast lipase (Candida cylindracea) . Studies with porcine pancreatic lipase were also conducted and the influence of various detergents and divalent cations on the assay was evaluated . The assay uses cis-parinaric acid (PnA), a naturally occurring fatty acid that has unique spectroscopic properties, and takes advantage of the reversible binding of fatty acids to bovine serum albumin (BSA) . Free PnA has an ultraviolet absorption peak at 321.2 nm . When PnA is bound to BSA, however, the peak shifts to 324.2 nm . The assay mixture contains 6 microM PnA, 1 microM BSA, 75 microM triolein, and 0.3 mM taurocholate in a 50 mM tris-HCl buffer with 1 microM EDTA . The release of oleic acid from triolein is monitored over time by measuring the ratio of optical densities (OD) at 319.0 and 329.0 nm . Initially, there is maximum binding of PnA to BSA, and the OD ratio is approximately 1.0 . Upon addition of lipase, PnA is displaced from the BSA by oleic acid released from triolein, and the OD ratio increases to a maximum of about 1.8 . However, when calcium is present in the reaction mixture an insoluble calcium-PnA complex forms, resulting in a progressive decrease in OD at both 319.0 and 329.0 nm . The kinetic assay described here is simple, rapid, sensitive, reproducible, inexpensive, and it can be adapted to measure the activity of a variety of calcium-independent lipases . Under similar assay conditions, activities for Candida cylindracea lipase obtained with this assay are similar to those obtained with 14C-labelled triolein.

Br J Dermatol, 1989 Jun, 120(6), 767 - 77
UVA sunbeds: tanning, photoprotection, acute adverse effects and immunological changes; Rivers JK et al.; The effects on 31 normal subjects following exposure to sunbeds containing UVA lamps with minimal UVB emission have been compared in a double-blind study with the effects on nine control subjects of a similar exposure course three times weekly for 4 weeks to sunbeds emitting visible light . On previously untanned areas, all those subjects on active treatment developed a mild tan; in tanned areas they all developed a moderate tan, while all control subjects developed a minimal to mild tan . The mean protection factor against later UVB-induced erythema was 3.2 +/- 0.3 after the active course and 1.6 +/- 0.2 among the controls . Significantly more frequent adverse cutaneous effects for active subjects were pruritus, erythema, freckling, burning sensation, dryness and polymorphic light eruption . Cutaneous Langerhans cell numbers, and blood CD3+ (pan T-cell) and CD4+ (helper T-cell) lymphocyte subsets were reduced in both active and control groups . CD8+ (cytotoxic/suppressor T-cell) counts were similarly but not significantly reduced in both groups . Pityrosporum yeast counts were significantly reduced in both groups . The changes found in both groups seem attributable to small amounts of UVB emission from both active and control lamps.

J Am Mosq Control Assoc, 1989 Jun, 5(2), 201 - 7
Improved techniques for rearing Anopheles freeborni; Fritz GN et al.; Techniques are described for mass rearing Anopheles freeborni . Eggs were incubated overnight at ca . 28 degrees C and then dried . Measured quantities of dried eggs were placed into styrofoam rings floating on the water surface of rearing trays . Water levels in larval rearing trays were kept shallow, and temperature was maintained with heat tapes at ca . 28 degrees C . Larvae were fed once a day on a slurry containing a 3:1:1:1 mixture of guinea pig chow, liver powder, yeast and hog chow . Pupation began on the 7th day after egg hatch, and pupae were harvested on the 8th, 9th and 10th days; ca . 1,700 pupae were harvested/tray . Adults emerged from 85% of the pupae, and about 40% were female . Individual males held in gallon-sized containers inseminated as many as 10 females . Although most sugar-fed males died within 2 weeks after emergence, over 35% of sugar-fed females survived for 3 weeks . Colonies were maintained on defibrinated bovine blood provided in natural membrane prophylactics . There were no significant differences in the number of blood-fed females or in the number of eggs they produced when mosquitoes were offered either guinea pigs or defibrinated bovine blood . Eggs were collected in plastic cups placed in cages . There was less than 6% mortality of eggs when these were dried and stored at 10 degrees C for 6 days.

Dis Colon Rectum, 1989 Jun, 32(6), 518 - 20
African histoplasmosis masquerading as carcinoma of the colon . Report of a case and review of literature; Khalil M et al.; A case of African histoplasmosis primarily affecting the large bowel--a very unusual manifestation--is reported . In the absence of classical clinical features, pathognomonic of large bowel histoplasmosis, the macroscopic appearance at surgery was confused with large-bowel cancer . The diagnosis was, therefore, retrospective and based on demonstration of large yeast cells of Histoplasma capsulatum var . duboisii in tissue sections of the lesions.

J Clin Lab Immunol, 1989 Jun, 29(2), 91 - 4
Neutrophil leucocyte function in primary hyperparathyroidism; Nordenstrom J et al.; Neutrophil leucocyte chemotaxis, phagocytosis and oxidative metabolism were measured in six patients with primary hyperparathyroidism (HPT) who underwent parathyroidectomy . The preoperative neutrophil chemotaxis value was 6.2 +/- 0.3 arb.U . and this decreased to 5.8 +/- 0.5 arb.U . (p less than 0.05) on the third postoperative day and to 5.4 +/- 0.3 arb.U . three weeks later . Serum calcium levels were 2.95 +/- 0.06 mmol/l preoperatively and decreased to 2.20 +/- 0.05 (p less than 0.05) and 2.34 +/- 0.05 mmol/l (p less than 0.05) on the third and 21st day after parathyroidectomy . Phagocytosis measured as the intracellular uptake of complement-opsonized yeast particles by neutrophils was not influenced by operation . In a control group, six patients undergoing hemithyroid ectomy due to microfollicular tumours were studied . In this group leucocyte chemotaxis, phagocytosis and serum calcium levels were not significantly different before and after surgery . Neutrophil oxidative metabolism, measured by the ability of the cells to reduce nitroblue tetrazolium (NBT), was similar in the HPT and control groups both before and after operation . The results indicate that HPT is associated with an abnormal leucocyte migration which is reversed after successful parathyroidectomy.

FEMS Microbiol Immunol, 1989 Jun, 1(6-7), 407 - 9
Lewis a blood group antigen of non-secretors: a receptor for candida blastospores; May SJ et al.; This study tested the hypothesis that the Lewis a blood group antigen found predominantly on the cells of non-secretors might be one of the receptors for Candida species . Binding of strain 3118C to epithelial cells from either secretor or non-secretor donors was not inhibited by treating the cells with anti-Lewis a or anti-Lewis b antisera . Binding of strain 3091 to non-secretor cells was inhibited by pretreating the cells with anti-Lewis a, but this was not observed for secretor cells . The results suggest that Lewis a might be one of the receptors for some yeast strains.

FEMS Microbiol Immunol, 1989 Jun, 1(6-7), 401 - 5
Non-secretion of blood group antigens and susceptibility to infection by Candida species; Thom SM et al.; One of the innate defences against superficial infections by Candida species appears to be the ability of an individual to secrete the water-soluble form of his ABO blood group antigens into body fluids . There was a significantly higher number of non-secretors (48.9%) among 174 patients with either oral or vaginal candida infections compared with the proportion of non-secretors in the local population (26.6%) . The protective effect afforded by the secretor gene might be due to the ability of glycocompounds in the body fluids of secretors to inhibit adhesins on the surface of the yeast . In attachment studies, preincubation of blastospores with boiled secretor saliva significantly reduced their ability to bind to epithelial cells . Non-secretor saliva did not reduce the binding and often enhanced the numbers of attached yeasts . Possible host-parasite interactions underlying the susceptibility of non-secretors to candida and other infections are discussed.

Eur J Biochem, 1989 Jun 1, 182(1), 27 - 36
Circular dichroic spectroscopy of membrane haemoproteins . The molecular determinants of the dichroic properties of the b cytochromes in various ubiquinol:cytochrome c reductases; Degli Esposti M et al.; The circular dichroism (CD) of dihaem cytochrome b from mitochondrial and bacterial ubiquinol:cytochrome-c reductase (bc1 complex) has been characterized . The dichroic properties of the yeast purified cyt b are very similar to those of the native cyt b within the mitochondrial bc1 complex . The CD spectra in the Soret region of the native cytochrome b present in all species studied show an intense bisignate Cotton effect having a zero-crossing wavelength close to the absorbance maximum . In preparations partially or completely depleted of the low-potential b haem (b1) the CD spectra exhibit a single positive Cotton effect resembling the corresponding absorption spectrum . This is particularly evident in the purified cytochrome b-562 from Rhodobacter sphaeroides R26, which contains only the high-potential b haem (bh) . These spectral features together with the reconstitution of the cytochrome b1 haem have been used to resolve the CD contribution of each haem to the CD spectra of cytochrome b . The mechanisms which might be responsible for the optical activity have been examined . It appears that the CD spectra of cytochrome b derive from both the mutual interaction of its two haems (giving rise to exciton coupling) and to the interaction of each haem with nearby aromatic residues, other than the pairs of histidines which coordinate the iron . The dipole coupling between haem and aromatic residues appears to be more important than exciton coupling in the CD spectra of oxidized b cytochromes and correlations have been made between the CD features and the proposed structure of cytochrome b.

J Immunol, 1989 Jun 1, 142(11), 3901 - 8
Recombinant soluble Fc epsilon receptor II (Fc epsilon RII/CD23) has IgE binding activity but no B cell growth promoting activity; Uchibayashi N et al.; We have undertaken the production of recombinant soluble Fc epsilon receptor II (Fc epsilon RII) as a secretory protein, but not as a cleavage product of membrane-bound receptor . Several plasmid constructs containing soluble receptor sequence were prepared . Only a chimeric gene containing the sequences encoding IL-6 signal peptide and the soluble moiety of Fc epsilon RII could be expressed in Xenopus laevis oocytes and CHO cells, resulting in the secretion of soluble Fc epsilon RII . The recombinant soluble Fc epsilon RII was also produced in the yeast expression system . The NH2-terminal sequence analysis of the chimeric gene product generated by oocytes demonstrated the correct cleavage of IL-6 leader sequence by a signal peptidase . Moreover, most of CHO cell and all of the yeast-derived recombinant molecules were products identical with the native B cell-derived soluble Fc epsilon RII . These recombinant products as well as the natural soluble receptor derived from a human B cell line could bind both human IgE and two different anti-Fc epsilon RII mAb and could competitively inhibit the binding of IgE to Fc epsilon RII-expressing cells . However, the recombinant soluble Fc epsilon RII and highly purified native molecules did not display any B cell growth-promoting activity.

Eur J Immunol, 1989 Jun, 19(6), 1123 - 9
Histone H1(0) mapping using monoclonal antibodies; Dousson S et al.; Monoclonal antibodies (mAb) to ox liver histone H1 degree were produced and characterized . Two sets of mice were immunized either with pure H1(0) or with an H1(0)-yeast tRNA complex . Eleven hybridomas of various clonal origin were selected . Typing of the antibodies indicated that all but three IgM belonged to the IgG1 class and contained kappa light chains . Immunoblotting experiments using peptides derived from H1(0) or H5 treated by various proteolytic agents (trypsin, N-bromosuccinimide, cyanogen bromide, acetic acid), revealed that nine of the mAb reacted with the globular part of H1(0) . More advanced characterization of the antigenic determinants allowed us to determine distinct regions within this globular part which are involved in the antigenic recognition . The peptopes could be subdivided into two groups . Three mAb bound to residues 24-27 and were specific for H1(0) . Six mAb bound to residues 27-30 and were specific for H1(0) except one of them which strongly cross-reacted with H5 and GH5 . Two mAb reacted with the entire histone H1(0) but failed to react with any of the peptides, suggesting that the corresponding epitope is a conformational antigenic determinant . In order to confirm the localization of the two distinct regions which are involved in the antigenic recognition, a synthetic decapeptide corresponding to the beginning of human H1(0) globular part (from residue 19 to residue 28) was synthesized . Inhibition experiments of the reaction between H1(0) and the various IgG1 mAb by increasing amounts of peptide-bovine serum albumin conjugates were then performed.

Biochem Biophys Res Commun, 1989 May 30, 161(1), 121 - 6
Characterization of ribonucleolytic activity of angiogenin towards tRNA; Lee FS et al.; Yeast tRNA is a convenient substrate for the assay of the ribonucleolytic activity of human angiogenin . The optimal pH, {NaCl}, and temperature for tRNA cleavage by angiogenin are approximately 6.8, 15-30 mM, and approximately 55 degrees C, respectively, as compared with approximately 8.0, 100-200 mM, and approximately 65 degrees C, respectively, for RNase A . Polyanions and metals both inhibit angiogenin and RNase A but to different extents.

Biochem J, 1989 May 15, 260(1), 231 - 5
Protoporphyrinogen oxidase as a molecular target for diphenyl ether herbicides; Matringe M et al.; Diphenyl ether herbicides induce an accumulation of protoporphyrin IX in plant tissues . By analogy to human porphyria, the accumulation could be attributed to decreased (Mg or Fe)-chelatase or protoporphyrinogen oxidase activities . Possible effects of acifluorfen-methyl on these enzymes were investigated in isolated corn (maize, Zea mays) etioplasts, potato (Solanum tuberosum) and mouse mitochondria, and yeast mitochondrial membranes . Acifluorfen-methyl was strongly inhibitory to protoporphyrinogen oxidase activities whatever their origins {concn . causing 50% inhibition (IC50) = 4 nM for the corn etioplast enzyme} . By contrast, it was roughly 100,000 times less active on (Mg or Fe)-chelatase activities (IC50 = 80-100 microM) . Our results lead us to propose protoporphyrinogen oxidase as a cellular target for diphenyl ether herbicides.

Nature, 1989 May 11, 339(6220), 147 - 9
A family of mitochondrial proteins involved in bioenergetICS and biogenesis; Schulte U et al.; The respiratory chain complexes of mitochondria consist of many different subunits, of which only a few partake directly in electron transport . The functions of the subunits that do not contain prosthetic groups are largely unknown . The cytochrome reductase complex of Neurospora crassa, for examine, consists of nine different subunits, of which the peripheral membrane proteins I and II (ref.3) that are located on the matrix side of the mitochondrial inner membrane are the largest subunits devoid of redox centres . Significantly, a cytochrome reductase fraction lacking these two subunits was inactive in electron transfer, and in yeast mutants with defective genes for either of the two subunits, assembly of the reductase is disrupted . Most mitochondrial proteins are imported into the mitochondrion as precursor proteins, and two proteins are necessary for cleaving their presequences, namely the matrix processing peptidase (MPP) and the processing enhancing protein (PEP), the latter strongly stimulating the activity of the former . Temperature-sensitive yeast mutants, which are affected in PEP or MPP, accumulate precursors at the nonpermissive temperature . We report here that subunit I of the cytochrome reductase can be grouped as members of the same protein family.

Cell, 1989 May 5, 57(3), 393 - 401
The cdc2 kinase is a nuclear protein that is essential for mitosis in mammalian cells; Riabowol K et al.; A homolog of the fission yeast cdc2-encoded protein kinase (p34) is a component of M phase promoting factor in Xenopus oocytes . The homologous kinase in human HeLa cells is maximally active during mitosis, suggesting a mitotic role in mammalian somatic cells . This has been directly investigated by microinjection of anti-p34 antibodies into serum-stimulated rat fibroblasts . DNA synthesis was unaffected but cell division was quantitatively blocked in injected cells . Injection of antibodies against p13suc1, a component of the p34 kinase complex, did not block mitosis but caused mitotic abnormalities resulting in cells containing multiple micronuclei in the subsequent interphase . p34 localized in the nucleus during interphase . During mitosis, a fraction tightly associated with centrosomes . p13 was more evenly distributed between the nucleus and cytoplasm . These observations demonstrate that cdc2 is a nuclear and centrosomal protein that is required for mitosis in mammalian cells.

Biochemistry, 1989 May 2, 28(9), 3948 - 54
Long-lived tryptophan fluorescence in phosphoglycerate mutase; Schauerte JA et al.; Phosphoglycerate mutase (PGM; EC 2.7.5.3) isolated from rat and rabbit muscle has been shown to possess an unusually long-lived fluorescence component when excited by ultraviolet light below 310 nm . On the basis of spectral and physical measurements, this 16.4 (+/- 0.2) ns fluorescence lifetime at room temperature is assigned to a tryptophan residue in an unusual environment . The emission profile of this long-lived tryptophan is red shifted from the other tryptophans of PGM by approximately 25 nm . PGM has been crystallized and sequenced from yeast where it has been shown to be a tetramer with 29K subunits . However, we have not been able to detect the existence of an unusually long-lived fluorescence component in the yeast isomer . The long fluorescence lifetime is lost upon denaturation of rabbit PGM and is partially restored upon introduction of the protein to a nondenaturing environment, suggesting the long lifetime is not the result of a covalent modification . The PGM molecule was studied by a number of techniques including time-resolved tryptophan fluorescence, quenching studies of tryptophan fluorescence, and enzyme activity studies . The long-lived fluorescence has been shown to be statistically quenched by Br-, I-, and Cu2+ in the submillimolar region while the acrylamide quenching shows the tryptophan is marginally accessible to solvent . Characterization of the long-lived fluorescence and its possible sources are discussed.

Plasmid, 1989 May, 21(3), 216 - 25
Detection of killer-independent dsRNA plasmids in Ustilago maydis by a simple and rapid method of extraction of dsRNA; Seroussi E et al.; A novel method for efficient and rapid isolation of dsRNA molecules was developed . The dsRNA content of Ustilago maydis was reexamined; two distinct dsRNA classes were identified . Class I includes the dsRNA segments reported earlier for U . maydis virus systems and class II includes unencapsidated dsRNA molecules that were barely detected by the conventional extraction methods despite their high titer . Segments of the class II, some of which are reported for the first time, were further characterized; all the segments are independent of the killer system and other encapsidated dsRNA molecules . These segments are cytoplasmically transmitted and, in sharp contrast with class I-encapsidated dsRNA segments, their relative copy number decreases rapidly while entering the stationary phase.

Mol Biol (Mosk), 1989 May-Jun, 23(3), 830 - 42
{Phylogenetic analysis of partial nucleotide sequences of 18S rRNA for 14 plant species}; Rakhimova GM et al.; The variable 260 base long region from the interior of 18S rRNA of 14 plant species was determined by chain termination method with the use of reverse transcriptase . The hairpin revealed in this region appeared to be conservative in all species compared . Thermodynamic stability of such hairpin is lower than of an alternative structure with different base pairing mode . From sequence data dendrograms were produced by clustering algorithms and by the compatibility method . In addition to the plant sequences these dendrograms included also the homologous regions from yeast and Xenopus 18S rRNAs . The compatibility method seems to be more reliable . Inferences were drawn on relations between gymnosperms and angiosperms, monocots and dicots on the bases of the analysis of this tree.

J Clin Immunol, 1989 May, 9(3), 229 - 41
In vitro immune responses to hepatitis B surface antigen (Pre-S2 and S) following remote infection by hepatitis B virus in humans; Cupps TR et al.; In this report we evaluate the human immune response to hepatitis B surface antigen (HBsAg) following remote infection with hepatitis B virus (HBV) . HBsAg-reactive lymphocytes can be readily demonstrated in the peripheral blood of individuals with established immunity following infection with HBV . In vitro stimulation with small doses of plasma-derived HBsAg, yeast-derived HBsAg (S region) or pre-S2 peptide will induce specific IgG to HBsAg (anti-HBs) in the absence of a polyclonal increase in total IgG . The pre-S2 peptide will stimulate, in a T cell-dependent fashion, the in vitro production of anti-HBs with specificity for the S domain . This anti-HBs production is mediated by pre-S2-stimulated soluble T-cell factors . Peripheral blood mononuclear cells from individuals with established immunity proliferate to the yeast-derived HBsAg but not to the plasma-derived HBsAg or pre-S2 peptide . The chronic HBsAg carriers do not produce anti-HBs following stimulation with HBsAg regardless of the source or component of antigen used . Different study protocols failed to demonstrate HBsAg-specific responses in the peripheral blood mononuclear cells of chronic carriers.

J Antimicrob Chemother, 1989 May, 23(5), 721 - 8
Clindamycin enhances murine delayed-type hypersensitivity and anti-candidal activity; Kitz DJ et al.; The contact sensitivity response of mice to dinitrofluorobenzene (DNFB), as determined by an epicutaneous painting and subsequent ear challenge assay, was enhanced by clindamycin administration . The optimal augmentation effect of clindamycin required its simultaneous administration at the time of DNFB skin sensitization . Clindamycin also was found to boost both in-vitro and in-vivo murine response to experimental infection with Candida lusitaniae . Intraperitoneal injection of 1-2 mg of the drug increased the clearance of yeast from organs after intravenous inoculation of mice . Clindamycin at concentrations of as low as 12.5 mg/l also increased the ability of cultured murine macrophages to kill the yeast without an increase in phagocytic activity.

Acta Cytol, 1989 May-Jun, 33(3), 337 - 40
Disseminated histoplasmosis diagnosed by fine needle aspiration biopsy of the adrenal gland . A case report; Anderson CJ et al.; A case of disseminated histoplasmosis diagnosed by fine needle aspiration (FNA) biopsy of the adrenal gland is reported for a 60-year-old man who presented with a 40-pound weight loss and abdominal computed tomography showing bilateral adrenal enlargement . FNA biopsy of the adrenal gland revealed clusters of macrophages with abundant cytoplasm containing the yeast forms of Histoplasma capsulatum . This case emphasizes that FNA is effective in diagnosing infectious as well as neoplastic conditions of the adrenal glands.

J Med Entomol, 1989 May, 26(3), 210 - 6
Factors influencing ingestion of particulate materials by mosquito larvae (Diptera: Culicidae); Rashed SS et al.; Relative ingestion rates of mosquito larvae, as indicated by the number of substrate-filled gut segments per unit time, were determined for Culex tarsalis Coquillett, Aedes aetypti L., and Anopheles albimanus Wiedemann . Among the three species, Ae . aegypti larvae were the most rapid feeders . F50 (median time for complete repletion of 50% of the larvae) was 61, 42, and 100 min for the three species feeding on a wheat flour suspension, respectively . Food particles with nutritive values (dried yeast, wheat flour, fishmeal, or dried blood) were ingested faster than inert particles (kaolin, talc, chalk, or charcoal) . The addition of aqueous yeast extracts containing phagostimulants accelerated the ingestion of inert particles . Increasing the concentration of inert particles did not increase ingestion rates . Larval age, water temperature, and starvation, but not larval density, influenced rates of ingestion . Younger instars were more rapid feeders than older instars . First instars of the three species filled their guts with wheat flour approximately two times faster than fourth instars . Increasing water temperature from 18 to 31 degrees C accelerated wheat flour ingestion by fourth instars of Cx . tarsalis, Ae . aegypti, and An . albimanus by factors of 1.9, 1.5, and 1.7, respectively . After starvation for 12 h, fourth instars of Cx . tarsalis and Ae . aegypti increased ingestion of wheat flour about 1.6 and 1.8 times, respectively . In contrast, starvation of An . albimanus larvae for the same period resulted in decreased wheat flour ingestion by 2.2 times when compared with unstarved larvae . These results indicare that, in addition to the chemical factors associated with the food substances, the physiological and environmatal conditions of the larvae play an important role in regulating the ingestion rate of suspended particles by mosquito larvae.

J Leukoc Biol, 1989 May, 45(5), 410 - 5
Interactions of pseudorabies virus with swine alveolar macrophages: effects of virus infection on cell functions; Iglesias G et al.; In order to assess the effect of pseudorabies virus (PRV) infection on the function of swine alveolar macrophages (AM), lung lavage cells were cultured, infected with one of six strains of PRV, and various activities were measured . Activity measurement included viability, phagocytosis of yeast, phagosome-lysosome fusion, phagocytosis of opsonized particles, and superoxide release . AM were infected with 5 x 10(-3) PFU/cell, and the comparative assessment of functions was performed at 18-20 h postinfection . Cell viability in PRV-infected cultures ranged from 79 to 94% of the viability in noninfected cultures . Phagocytosis of yeast was significantly reduced only in the AM cultures infected with the strain S-62 . Phagosome-lysosome fusion was depressed in cultures infected with the strains S-62, 4892, 3816, and BUK . The phagocytosis of opsonized sheep red blood cells showed significant differences between noninfected and PRV-infected cultures in all cases except cultures infected with the strain PRV-C . The O2 release after stimulation with opsonized zymosan was significantly reduced in all the PRV-infected cultures . The effect of PRV infection on AM functions that are related to the bacterial activity of such cells suggests that PRV-induced AM dysfunction might have a role in the increased susceptibility of PRV-infected pigs to bacterial pneumonia.

J Med Microbiol, 1989 May, 29(1), 51 - 4
Candida species and C . albicans biotypes in women attending clinics in genitourinary medicine; Odds FC et al.; Yeasts were isolated from two or more anatomical sites in 198 women attending genitourinary clinics on at least two occasions . The yeast biotypes isolated concurrently from the vagina and urethra were the same in 138 (99%) of 140 instances, and 94% of 124 concurrent genital and anal isolates were of matching types, whereas only 75% of concurrent genital and oral isolates were of the same type . Mixtures of Candida spp . or C . albicans biotypes were encountered only five times among 545 yeast-positive samples . In instances where Candida spp . were isolated at successive times from the same site in a patient, the same yeast type was encountered on 97 (87%) of 112 occasions when the interval between samples was less than 15 weeks, and on 19 (66%) of 29 occasions when the interval was 15 weeks or more . These data indicate a tendency to carriage of phenotypically consistent types of Candida among most women attending genitourinary clinics.

Mol Cell Biol, 1989 May, 9(5), 2273 - 8
Remarkable intron and exon sequence conservation in human and mouse homeobox Hox 1.3 genes; Tournier-Lasserve E et al.; A high degree of conservation exists between the Hox 1.3 homeobox genes of mice and humans . The two genes occupy the same relative positions in their respective Hox 1 gene clusters, they show extensive sequence similarities in their coding and noncoding portions, and both are transcribed into multiple transcripts of similar sizes . The predicted human Hox 1.3 protein differs from its murine counterpart in only 7 of 270 amino acids . The sequence similarity in the 250 base pairs upstream of the initiation codon is 98%, the similarity between the two introns, both 960 base pairs long, is 72%, and the similarity in the 3' noncoding region from termination codon to polyadenylation signal is 90% . Both mouse and human Hox 1.3 introns contain a sequence with homology to a mating-type-controlled cis element of the yeast Ty1 transposon . DNA-binding studies with a recombinant mouse Hox 1.3 protein identified two binding sites in the intron, both of which were within the region of shared homology with this Ty1 cis element.

Acta Trop, 1989 May, 46(3), 191 - 203
Isolation and cultivation in vitro to the infective, metacyclic stage of Trypanosoma (Nannomonas) simiae from Glossina morsitans submorsitans; Dukes P et al.; Two separate trypanosome isolations were made from a single Nannomonas-infected Glossina morsitans submorsitans from The Gambia . Inoculation of a piglet with the infected hypopharynx produced an infection with Trypanosoma simiae . DNA was isolated from the bloodstream forms to prepare a probe specific for this species . Trypanosomes isolated from the fly midgut were frozen in liquid nitrogen and then cultivated in vitro . Amplification of this population and elimination of a yeast contaminant were achieved by two passages through laboratory G . m . morsitans . Further cultivation in vitro resulted in the production of epimastigotes and, later, metacyclic forms . Two pigs inoculated with cultivated metacyclic forms developed infections with atypical, relapsing parasitaemias and extended survival time . Neither the metacyclic forms, nor bloodstream forms derived from them, infected calves . The identity of various stages of the in vitro cultivated, procyclic-derived stock was confirmed morphologically and with the T . simiae-specific DNA probe.

Mol Cell Biol, 1989 May, 9(5), 2042 - 9
Expression of alpha- and beta-tubulin genes during dimorphic-phase transitions of Histoplasma capsulatum; Harris GS et al.; Recent investigations have confirmed the presence of one alpha-tubulin gene (TUB1) and one beta-tubulin gene (TUB2) in the dimorphic fungus Histoplasma capsulatum . In the present study, Northern blot (RNA blot) analyses revealed multiple alpha-tubulin transcripts and a single beta-tubulin transcript in the yeast and mycelial phases of the high-virulence 217B strain and low-virulence Downs strain . S1 nuclease protection assays demonstrated one initiation start site and two major stop sites for the TUB1 transcripts, suggesting that variations in 3' processing generate the alpha-tubulin messages of 2.5 and 2.0 kilobases . Dot blot hybridization experiments indicated that tubulin gene expression is developmentally regulated during the dimorphic phase transitions . alpha- and beta-tubulin mRNAs increased six- to eightfold during the yeast-to-mycelium conversion and decreased two- to threefold during the reverse transition . These changes in tubulin mRNA content coincided with major morphological events associated with H . capsulatum development . Western blots (immunoblots) of H . capsulatum yeast-specific proteins resolved by two-dimensional gel electrophoresis demonstrated a single alpha- and a single beta-tubulin isoform . Multiple tubulin polypeptides expressed in mycelia are probably products of posttranslational modifications.

J Clin Microbiol, 1989 May, 27(5), 1097 - 8
Comparison of four charcoal media for the isolation of Bordetella pertussis; Hoppe JE et al.; Charcoal-horse blood agar with 40 micrograms of cephalexin per ml, charcoal-horse blood agar with 3 micrograms of lincomycin per ml, charcoal agar with 3 micrograms of lincomycin per ml, and Legionella (buffered charcoal-yeast extract) agar with 3 micrograms of lincomycin per ml were compared for isolation of Bordetella pertussis . Charcoal-horse blood agar with 40 micrograms of cephalexin per ml gave the best results, with a B . pertussis recovery rate of 100% . Growth was most rapid and the mean number of colonies was highest on this agar, and growth of pharyngeal flora was completely suppressed.

Exp Parasitol, 1989 May, 68(4), 443 - 9
Characterization of a protein fraction containing cytochromes b and c1 from mitochondria of Leishmania tarentolae; Shaw JM et al.; A soluble red band fraction was obtained from Leishmania tarentolae cells by sucrose gradient sedimentation of a Triton X-100 lysate . Spectral analysis indicated that cytochrome b was present in the red band: the reduced minus oxidized difference spectra revealed absorption maxima at 562,527, and 431 nm at room temperature and 562, 530, and 422 nm at 77K . In addition, a 28-kDa protein was identified in this fraction which retained heme-associated peroxidase activity even after denaturation on SDS-polyacrylamide gels . The amino acid composition of this protein showed a strong similarity to cytochrome c1 of both bovine and yeast.

J Am Acad Dermatol, 1989 May, 20(5 Pt 1), 770 - 3
Association of Pityrosporum orbiculare (Malassezia furfur) with seborrheic dermatitis in patients with acquired immunodeficiency syndrome (AIDS); Groisser D et al.; The possible causative role of the yeastlike fungus Pityrosporum (Malassezia) orbiculare in the pathogenesis of seborrheic dermatitis in patients with and without acquired immunodeficiency syndrome (AIDS) has been discussed but not resolved . Ten patients with AIDS-related seborrheic dermatitis were studied for the presence of Pityrosporum organisms . On the basis of a quantitative correlation between numbers of yeast cells adherent to and extruded from keratinocytes and the clinical severity of seborrheic dermatitis, an association, if not a causative role, for Pityrosporum is strongly suggested in seborrheic dermatitis in patients with AIDS . This association was further strengthened by the marked clinical response to ketoconazole in two patients with a concomitant decrease in the number of Pityrosporum cells per keratinocyte.

Trends Biochem Sci, 1989 May, 14(5), 172 - 5
The oncogene jun and nuclear signalling; Vogt PK et al.; Jun is a transcription factor that can also induce oncogenic transformation . Its DNA-binding domain is conserved from yeast to man and shows homology to several other transcriptional regulators . Jun dimerizes with the fos protein through an alpha-helical domain termed the leucine zipper, and the jun-fos heterodimers bind to DNA and regulate transcription of numerous specific unlinked genes.

Biochem J, 1989 May 1, 259(3), 847 - 53
Immunochemical characterization of NADPH-cytochrome P-450 reductase from Jerusalem artichoke and other higher plants; Benveniste I et al.; Polyclonal antibodies were prepared against NADPH-cytochrome P-450 reductase purified from Jerusalem artichoke . These antibodies inhibited efficiently the NADPH-cytochrome c reductase activity of the purified enzyme, as well as of Jerusalem artichoke microsomes . Likewise, microsomal NADPH-dependent cytochrome P-450 mono-oxygenases (cinnamate and laurate hydroxylases) were efficiently inhibited . The antibodies were only slightly inhibitory toward microsomal NADH-cytochrome c reductase activity, but lowered NADH-dependent cytochrome P-450 mono-oxygenase activities . The Jerusalem artichoke NADPH-cytochrome P-450 reductase is characterized by its high Mr (82,000) as compared with the enzyme from animals (76,000-78,000) . Western blot analysis revealed cross-reactivity of the Jerusalem artichoke reductase antibodies with microsomes from plants belonging to different families (monocotyledons and dicotyledons) . All of the proteins recognized by the antibodies had an Mr of approx . 82,000 . No cross-reaction was observed with microsomes from rat liver or Locusta migratoria midgut . The cross-reactivity generally paralleled well the inhibition of reductase activity: the enzyme from most higher plants tested was inhibited by the antibodies; whereas Gingko biloba, Euglena gracilis, yeast, rat liver and insect midgut activities were insensitive to the antibodies . These results point to structural differences, particularly at the active site, between the reductases from higher plants and the enzymes from phylogenetically distant plants and from animals.

J Invertebr Pathol, 1989 May, 53(3), 358 - 60
Immunological separation of Entomophthorales genera; Toriello C et al.; Exoantigens from Erynia neoaphidis, Conidiobolus major, C . thromboides, C . obscurus, Zoophthora radicans, and Basidiobolus ranarum were obtained from culture filtrates of fungal material grown in a yeast extract, peptone dialysate, dextrose medium and were tested against specific hyperimmune antisera prepared from E . neoaphidis, C . major, C . thromboides, and B . ranarum by the immunodiffusion technique . Specific precipitins were observed for E . neoaphidis and B . ranarum, while cross-reactions were detected among C . major, C . thromboides, and C . obscurus . The results suggest that genera of Entomophthorales can be easily separated by this simple immunological procedure.

Somat Cell Mol Genet, 1989 May, 15(3), 237 - 44
Sequence analysis of mouse mitochondrial chloramphenicol-resistant mutants; Howell N et al.; The nucleotide sequences of the 3' halves of the mitochondrial 16S rRNA genes from four independent mouse chloramphenicol-resistant (CAP-R) mutants were determined . Each contained a different, single base change that encodes the mutational phenotype . The mitochondrial rRNA gene from the SVA31 CAP-R mutant contains a G-to-A transition at nucleotide 2161 of the noncoding strand; the SVIS CAP-R mutant, a G-to-A transition at position 2375; the LA9 CAP-R mutant, an A-to-T transversion at position 2379; and the SVT2 CAP-R mutant, a T-to-C transition at position 2433 . Three of these CAP-R mutants appear to be heteroplasmic as the mtDNA populations contain both wild-type and mutant copies of the rRNA gene . The SVIS CAP-R mutation has not been observed in other mammalian CAP-R mutants, although it occurs at a site homologous to one of the yeast mitochondrial CAP-R mutations . Based upon the locations of the mutated sites within the 16S rRNA, and their proximity to previously analyzed sites of mutations conferring increased inhibitor resistance, all these mutations occur within the ribosomal RNA peptidyltransferase domain . These results provide an explanation for the pleiotropic nature of mitochondrial CAP-R mutations in mammalian cells, particularly the observations that some of the mutant lines are partially respiration deficient.

Cell, 1989 Apr 21, 57(2), 253 - 63
Purification of MPF from starfish: identification as the H1 histone kinase p34cdc2 and a possible mechanism for its periodic activation; Labbe JC et al.; MPF extracted from starfish oocytes copurifies with an M phase-specific H1 histone kinase encoded by a homolog of the fission yeast cell cycle control gene cdc2+ . The most purified preparations contain p34cdc2 as the only major protein . Activation of the p34cdc2 kinase is correlated with appearance of the MPF activity both in vivo and in vitro . The increase in protein kinase activity is associated with p34cdc2 dephosphorylation and the decrease in protein kinase activity on leaving M phase with rephosphorylation . Microinjection of a peptide perfectly conserved in p34cdc2 from yeast to humans induces meiotic maturation, suggesting that an inhibitory component in G2 arrested oocytes interacts with this region of the p34cdc2 kinase . We propose that initiation of M phase is brought about by the dephosphorylation of p34cdc2, leading to increase in its protein kinase activity.

Science, 1989 Apr 21, 244(4902), 362 - 4
An assay for circulating antibodies to a major etiologic virus of human non-A, non-B hepatitis; Kuo G et al.; A specific assay has been developed for a blood-borne non-A, non-B hepatitis (NANBH) virus in which a polypeptide synthesized in recombinant yeast clones of the hepatitis C virus (HCV) is used to capture circulating viral antibodies . HCV antibodies were detected in six of seven human sera that were shown previously to transmit NANBH to chimpanzees . Assays of ten blood transfusions in the United States that resulted in chronic NANBH revealed that there was at least one positive blood donor in nine of these cases and that all ten recipients seroconverted during their illnesses . About 80 percent of chronic, post-transfusion NANBH (PT-NANBH) patients from Italy and Japan had circulating HCV antibody; a much lower frequency (15 percent) was observed in acute, resolving infections . In addition, 58 percent of NANBH patients from the United States with no identifiable source of parenteral exposure to the virus were also positive for HCV antibody . These data indicate that HCV is a major cause of NANBH throughout the world.

J Mol Biol, 1989 Apr 20, 206(4), 567 - 77
Nucleotide sequences of Caenorhabditis elegans core histone genes . Genes for different histone classes share common flanking sequence elements; Roberts SB et al.; We have determined the nucleotide sequence of core histone genes and flanking regions from two of approximately 11 different genomic histone clusters of the nematode Caenorhabditis elegans . Four histone genes from one cluster (H3, H4, H2B, H2A) and two histone genes from another (H4 and H2A) were analyzed . The predicted amino acid sequences of the two H4 and H2A proteins from the two clusters are identical, whereas the nucleotide sequences of the genes have diverged 9% (H2A) and 12% (H4) . Flanking sequences, which are mostly not similar, were compared to identify putative regulatory elements . A conserved sequence of 34 base-pairs is present 19 to 42 nucleotides 3' of the termination codon of all the genes . Within the conserved sequence is a 16-base dyad sequence homologous to the one typically found at the 3' end of histone genes from higher eukaryotes . The C . elegans core histone genes are organized as divergently transcribed pairs of H3-H4 and H2A-H2B and contain 5' conserved sequence elements in the shared spacer regions . One of the sequence elements, 5' CTCCNCCTNCCCACCNCANA 3', is located immediately upstream from the canonical TATA homology of each gene . Another sequence element, 5' CTGCGGGGACACATNT 3', is present in the spacer of each heterotypic pair . These two 5' conserved sequences are not present in the promoter region of histone genes from other organisms, where 5' conserved sequences are usually different for each histone class . They are also not found in non-histone genes of C . elegans . These putative regulatory sequences of C . elegans core histone genes are similar to the regulatory elements of both higher and lower eukaryotes . The coding regions of the genes and the 3' regulatory sequences are similar to those of higher eukaryotes, whereas the presence of common 5' sequence elements upstream from genes of different histone classes is similar to histone promoter elements in yeast.

Nature, 1989 Apr 13, 338(6216), 589 - 90
Two functionally different regions in Fos are required for the sequence-specific DNA interaction of the Fos/Jun protein complex; Neuberg M et al.; THE products of the cellular and retroviral fos genes associate with other nuclear proteins, among them the transcription factor AP1/Jun (see ref . 3 for a review) . The Fos/Jun complex binds to a specific symmetrical DNA recognition sequence (termed TRE), thus stimulating transcription of the respective gene . Here, we show that two distinct regions in Fos are required for the formation of a Fos/Jun/TRE complex . These are the leucine zipper, involved in the association with Jun, and a directly adjacent basic region . Specific amino-acid substitutions in this basic, presumably alpha-helical, region abolish the interaction of Fos/Jun with the TRE but not the association of the two proteins . The functionally crucial amino acids are located in a region of Fos which is structurally similar to the putative DNA-binding sites in Jun and in the yeast transcriptional activator GCN4 (refs 15 and 16).

J Parasitol, 1989 Apr, 75(2), 261 - 9
Phagocytic activity of hemocytes of M-line Biomphalaria glabrata snails: effect of exposure to the trematode Echinostoma paraensei; Noda S et al.; The phagocytic activity of hemocytes from 6-8-mm M-line Biomphalaria glabrata snails was studied in an in vitro assay using glutaraldehyde-fixed sheep erythrocytes (SRBC) as target cells . For individual snails, the percentage of hemocytes ingesting SRBC during a 1-hr interval, termed the phagocytic activity index (PAI), was determined . Hemocytes from snails infected for 1 day with Echinostoma paraensei had a slightly elevated PAI, but at both 8 and 30 days postexposure (DPE), hemocytes from infected snails had a significantly lower PAI than controls . Hemocytes taken from snails at 8 DPE also had a low PAI using rabbit erythrocytes and yeast as target cells . The low PAI at 8 DPE is attributed to the presence of large numbers of poorly spreading hemocytes with low phagocytic activity . Hemocytes from snails with 30-day infections were well spread but nonetheless had a low PAI . The presence of plasma from 8-day infected snails did not alter the PAI of hemocytes from control snails, nor was the PAI of hemocytes from infected snails changed by plasma from control snails . SRBC preincubated for 60 min in plasma from various groups of M-line snails did not elicit an increase in PAI when presented to hemocytes from control snails; in some cases, as with plasma from 6-8-mm control snails, such preincubation significantly reduced the PAI below levels obtained using SRBC preincubated in culture medium . As compared to hemocytes from snails with normally developing, 8-day-old intraventricular sporocysts (IS), hemocytes from snails exposed to infection but subsequently lacking IS had a significantly higher PAI.(ABSTRACT TRUNCATED AT 250 WORDS)

Chem Pharm Bull (Tokyo), 1989 Apr, 37(4), 1068 - 70
Further studies on the pharmacological effect of the anti-inflammatory compound, bis{2-(E-2-octenoylamino)ethyl} disulfide; Mimura T et al.; Further studies on the pharmacological effect of orally administered bis{2-(E-2-octenoyl-amino)ethyl} disulfide (compd . I-3) was examined by using several experimental models in vivo . Compound I-3 showed analgesic activity, inhibiting both acetic acid- and acetylcholine-induced writhings in mice . This compound also showed antipyretic activity against yeast-induced fever in rats, and significantly inhibited arachidonic acid-induced mortality in mice . However, it had no effect on serotonin-induced paw edema formation or platelet activating factor-acether-induced mortality in mice . The effects of compd . I-3 are suggested to be due to inhibition of prostaglandin biosynthesis.

Genitourin Med, 1989 Apr, 65(2), 106 - 8
Survival of Trichomonas vaginalis in human semen; Daly JJ et al.; Although exposure of Trichomonas vaginalis to human semen is of short duration, any effect that this fluid may have on the urogenital protozoon could affect its transmission, especially if only few trichomonads are present . Small numbers of parasites (about 2500/ml semen) incubated in semen from different donors at 37 degrees C, were found to survive or grow for up to 12 hours in all samples and for up to 24 hours in most . Survival and growth of T vaginalis in semen most resembled that found in Diamond's trypticase, yeast extract, and maltose (TYM) medium without serum supplement, rather than in complete TYM medium and phosphate buffered saline . Contrary to previous reports, semen did not inhibit the survival of T vaginalis, and the presence of trichomonads did not alter motility or numbers of spermatozoa up to 24 hours . The data suggest that semen provides a favourable milieu for transmitting trichomonads.

Mycoses, 1989 Apr, 32(4), 183 - 6
Trichosporon beigelii infection in an immunocompromised host; Payne AL et al.; A case of Trichosporon beigelii infection in a patient with non-Hodgkins lymphoma that illustrates some of the associated diagnostic and chemotherapeutic problems, is described . Despite prolonged isolation of the yeast from blood cultures, the patient recovered from the infection after treatment with amphotericin B and flucytosine . Presenting features, diagnosis and monitoring of antifungal therapy in renal failure are discussed.

Genes Dev, 1989 Apr, 3(4), 572 - 83
xlgv7: a maternal gene product localized in nuclei of the central nervous system in Xenopus laevis; Miller M et al.; The Xenopus oocyte nucleus (GV) is a storehouse for a large number of proteins that are used during early development . We have cloned and characterized a cDNA coding for a maternal gene product that is localized in the GV and then becomes highly enriched in the nuclei of the central nervous system (CNS) of tadpoles and adult frogs . This cDNA (xlgv7) is 2.1 kb and hybridizes to a 2.4-kb RNA species on Northern blots . Southern blots of genomic DNA suggest that this gene is a member of a multigene family . The cDNA sequence reveals a long open reading frame (ORF) of 1773 nucleotides, with a putative nuclear targeting signal (Glu Arg Arg Lys Lys Lys Thr) at the extreme carboxyl terminus and an internal histidine (His)-rich region with a repeated conserved amino acid sequence between His pairs . The significance of this region is unclear, but the protein is a DNA-binding protein, and it is possible that this region is involved in this function . The xlgv7 protein also possesses a putative nucleotide-binding consensus sequence that is similar to the bacterial RecA and RecB and yeast RAD proteins . Protein xlgv7 exists as several isotypes that exhibit developmental and cell-specific changes during development . Northern blot analysis of the abundance of the xlgv7 mRNA shows an accumulation following neural induction at stages 15-16 . There is a transient expression of the mRNA in the gut of tadpoles . In the adult, the mRNA is highly enriched in the brain and is absent or in very low abundance in other tissues . Immunohistochemical analysis of the protein shows that the protein is localized in the nuclei of the brain cells . We conclude that the xlgv7 gene product is a maternal protein that may serve several important functions, one of which may be in the development and maintanance of the CNS.

Genomics, 1989 Apr, 4(3), 376 - 83
Genomic organization of human 5 S rDNA and sequence of one tandem repeat; Little RD et al.; An organization of human 5 S rDNA repeats is inferred from Southern analyses of restriction digests of genomic DNA fractionated by pulsed-field and conventional gel electrophoreses . A single unit of 2.2 kb is repeated approximately 90 times within a 200-kb fragment (defined by enzymes that do not cleave within individual units, i.e., EcoR1, BglII, HindIII, and PvuII); a comparable number of 5 S sequences are scattered elsewhere in the genome . A lambda clone containing six complete 5 S repeats was obtained from a human placental DNA library . One repeat contains 2231 bp and includes poly(dG-dT).(dC-dA), tracts of polypyrimidine, and an Alu sequence in the spacer region . Also, 5-S-hybridizing clones, containing DNA inserts with an average size of 250 kb, have been obtained as yeast artificial chromosomes . Thus far, four clones have been partially characterized and shown to be 5 S sequences from loci separate from the tandem repeat units.

Proc Natl Acad Sci U S A, 1989 Apr, 86(8), 2563 - 7
Complete amino acid sequence of rat brain hexokinase, deduced from the cloned cDNA, and proposed structure of a mammalian hexokinase; Schwab DA et al.; The complete amino acid sequence for the type I isozyme of hexokinase from rat brain has been deduced from the nucleotide sequence of cloned cDNA . The nucleotide sequence of 91 bases in the 5' untranslated region as well as that of the entire 3' untranslated region preceding the poly(A) sequence have also been determined . The N- and C-terminal halves of brain hexokinase show extensive sequence similarity to each other and to yeast hexokinase . These results provide direct support for the proposal that the mammalian hexokinases of approximately 100 kDa have evolved by a process of duplication and fusion of a gene encoding an ancestral hexokinase similar to the yeast enzyme of approximately 50 kDa . Taking this similarity in sequence to indicate basic similarity in structure between the N- and C-terminal regions of brain hexokinase and the yeast enzyme, a proposed structure for the mammalian hexokinase has been developed by fusing two molecules of yeast hexokinase, whose structure has previously been determined by x-ray crystallographic studies . Various features of the model are shown to be consistent with experimental observations bearing on the structure of the brain enzyme.

Revmatologiia (Mosk), 1989 Apr-Jun, (2), 30 - 5
{Anti-deoxyribonucleic acid antibodies in podagra}; Nikolenko IuI et al.; Antibodies to native (n) and denatured (d) DNA were studied in 70 patients with primary gout and 40 experimental animals with hyperuricemia . Titres of anti-nDNA in patients with gout considerably exceeded similar indices in donors and were in inverse relationship with the severity of arthritis and markedness of morphological manifestations of renal affection . Anti-nDNA directly correlated with the count of B-lymphocytes in the blood and indirectly with other immunological indices . Anti-dDNA directly correlated with the count of B-cells, reaction of lymphocytes to Kon-A and the titres of antirenal antibodies but were in inverse relationship with the content of uric acid and IgG in the blood serum . Long-term feeding of rats with yeast autolysate with ammonium molybdate and inosine was accompanied by the development of hyperuricemia, an abrupt drop in the level of DNA in the plasma and by a rise in the level of anti-DNA antibodies.

Arch Invest Med (Mex), 1989 Apr-Jun, 20(2), 123 - 7
Excystation and culture of Giardia spp . from human source; Ponce Macotela M et al.; In this paper, we report the hatching of Giardia spp . trophozoites from purified cysts harvested in Sheather's solution from human feces . Isolated cysts were induced by acidic Hank's Balanced Salt Solution (HBSS), incubated at 37 degrees C for 30 minutes, concentrated by centrifugation and washed with pH 7.5 HBSS . The cysts were placed in 8 ml . culture medium (Pancreatic digest of casein and yeast extracts) supplemented with human serum and bile in borosilicate glass tubes with screw caps . Trophozoite excystation was scanned in a Commandon chamber filled with the suspension, sealed with a cover glass and vaseline . The trophozoite emergence was recorded with phase contrast in a photomicroscope . The excystation began within the first 3 minutes, after inoculation into medium . The trophozoites emerged trough a small hole at one end of the cyst and simultaneously began cellular division, the whole process required 5-30 minutes . The trophozoites attached themselves to the walls of the culture tube, permitting the replacement and removal of contaminants without dislodging them from the tube walls . The isolates MM-INP and MG-INP have been maintained at 37 degrees C for more than six months.

J Neurosci Res, 1989 Apr, 22(4), 384 - 9
Developmental and regional expression of three new members of the ras-gene family in the mouse brain; Ayala J et al.; We have examined the expression in the mouse nervous system of three new members of the ras protooncogene family: rab1, rab2, and rab3 . Each of these genes was transcribed into messenger RNAs with different molecular weights . These transcripts has specific developmental and regional patterns of expression . In particular, for the three genes, the ratio between the heavy and light mRNAs depended strongly on developmental stage and brain region . The use of pure neuronal and glial cultures revealed that the high molecular weight transcripts were enriched in neurons and that, in the case of rab2 and rab3, their expression increased with neuronal differentiation . These results are discussed considering the sequence identities between these genes and the yeast YPTI and sec-4 genes, which are known to be implicated in post-Golgi vesicular transport and cytoskeletal stabilization . We propose that the rab genes might be of importance in the regulation of these two processes within the developing and adult nervous system.

Biochem J, 1989 Apr 1, 259(1), 145 - 52
A super-secondary structure predicted to be common to several alpha-1,4-D-glucan-cleaving enzymes; MacGregor EA et al.; Predictions of protein secondary structure are used with amino acid sequence alignments to show that the N-terminal domains of cyclodextrin glucanotransferases and a yeast alpha-glucosidase may have the same super-secondary structure as alpha-amylases, i.e . an (alpha/beta)8-barrel fold . Sequence similarities provide evidence that glucanotransferases, and possibly the glucosidase, are, like alpha-amylases, Ca2+-containing enzymes . The relationship between substrate specificity and the nature of the amino acid residues proposed at the active site is discussed for the transferases and alpha-glucosidase . A set of three programs for an Apple IIe computer to carry out the calculations described by Garnier, Osguthorpe & Robson {(1978) J . Mol . Biol . 120, 97-120} and a set of four programs for an Apple IIe computer to carry out the calculations described by Levin, Robson & Garnier {(1986) FEBS Lett . 205, 303-308} have been deposited as Supplementary Publication SUP 50149 (25 pages) at the British Library Document Supply Centre, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem . J . (1989) 257, 5.

Biochimie, 1989 Apr, 71(4), 497 - 503
Lysine degradation in Candida . Characterization and probable role of L-norleucine-leucine, 4-aminobutyrate and delta-aminovalerate:2-oxoglutarate aminotransferases; Der Garabedian PA et al.; Three enzymes partially purified that catalyze respectively the transamination of L-norleucine, 4-aminobutyrate and delta-aminovalerate with alpha-ketoglutarate as aminoacceptor were characterized and isolated from L-lysine adapted cell of Candida guilliermondii var . membranaefaciens . The transaminases have a maximum activity in the pH range of 7.8-8.5 and at 55 degrees C, 45 degrees C and 40 degrees C respectively . alpha-Ketoglutarate and to a lesser extent pyridoxal-5'-phosphate were effective protecting agents against rise in temperature . The enzymes exhibit absorption maximum at 280 nm, 330 nm and 410 nm . The fact that L-norleucine-leucine aminotransferase, 4-aminobutyrate aminotransferase and delta-aminovalerate aminotransferase are strongly induced by growing the yeast Candida on L-lysine suggests new hypothetic pathways for the catabolism of L-lysine where the main substrate of each aminotransferase could be an intermediary metabolite.

Biol Trace Elem Res, 1989 Apr-May, 20(1-2), 59 - 65
Selenium supplementation of symptomatic human immunodeficiency virus infected patients; Olmsted L et al.; The mean whole blood selenium levels in male San Diego, CA patients with acquired immune deficiency syndrome (AiDS) are 0.123 +/- 0.030 micrograms/mL (n = 24), and 0.126 +/- 0.038 micrograms/mL (n = 26) in patients with AIDS-related complex (ARC), compared to 0.195 +/- 0.020 micrograms/mL (n = 28) in San Diego healthy controls (males) . To establish whether intestinal absorption of dietary selenium is impaired in AIDS or ARC, a supplementation trial was conducted in which 19 symptomatic HIV-antibody positive male patients with AIDS or ARC were taking 400 micrograms of selenium/d in form of selenium yeast for up to 70 d . The mean whole blood Se levels increased to 0.28 +/- 0.08 micrograms/mL after 70 d of supplementation, the selenium supplements were well tolerated . A rationale for adjuvant selenium supplementation of symptomatic and asymptomatic HIV carriers is proposed.

Mol Pharmacol, 1989 Apr, 35(4), 428 - 32
Transfer RNA is cleaved by activated bleomycin; Magliozzo RS et al.; Activated bleomycin is shown for the first time to cleave tRNA in a specific and dose-dependent manner . Adenine and uracil are released in the reaction . Bleomycin and Fe(III)-bleomycin bind to yeast tRNAPhe) in analogy with the known behavior of the drug with B-DNA.

Nature, 1989 Mar 30, 338(6214), 438 - 40
Identification of the long ubiquitin extension as ribosomal protein S27a; Redman KL et al.; Two proteins of unknown function are encoded by 3' in-frame extensions of ubiquitin genes . The polypeptides are synthesized as an additional 52 or 76-80 amino acids on the C terminus of ubiquitin, an unusual arrangement conserved in man, yeast and plants (J . Callis and R . Vierstra, personal communication) . Although not homologous to each other or to ubiquitin, both extension proteins are highly basic and contain patterns of cysteine and histidine similar to those proposed to form 'zinc fingers' . The longer C-terminal extension protein (CEP80) is 30% lysine and arginine and, when denatured, behaves like a small cationic protein . Its properties after isolation in physiological conditions, however, suggested that CEP80 is part of an RNA-protein complex . Using the antibodies that confirmed the presence of CEP80 in eukaryotic cells, we show here that the protein is located on ribosomes . Immunoblotting of rat 40S subunit proteins specifically identifies CEP80 as ribosomal protein S27a.

J Biol Chem, 1989 Mar 25, 264(9), 4957 - 63
Mouse primase p49 subunit molecular cloning indicates conserved and divergent regions; Prussak CE et al.; Primase is a specialized RNA polymerase that synthesizes RNA primers for initiation of DNA synthesis . A full cDNA clone of the p49 subunit of mouse primase, a heterodimeric enzyme, has been isolated using a primase p49-specific polyclonal antibody to screen a lambda gt11 mouse cDNA expression library . The cDNA indicated the subunit is a 417-amino acid polypeptide with a calculated molecular mass of 49,295 daltons . The p49 mRNA is approximately 1500 nucleotides long with a 5'-untranslated region of 74 nucleotides and a 3'-untranslated region of 200 nucleotides . Comparison with a similar sized primase subunit from yeast showed highly conserved amino acid sequences in the N-terminal halves of the polypeptides and included a potential metal-binding domain suggesting the functional importance of this region for DNA binding . In contrast, the 3' portion of the cDNA has rapidly diverged in nucleotide sequence, as primase mRNA can be detected in mouse and rat cells with a 3' probe (including coding and noncoding) but not in RNA from hamster or human cells . A full-length cDNA probe detected mRNA from hamster and human cell lines, indicating a conserved 5' portion and divergent 3' region of the expressed gene . The rapid divergence may be related to the species-specific protein interactions found for the DNA polymerase alpha-primase complex . The mRNA is detected in proliferating but not in quiescent cells consistent with its function in DNA replication.

Nature, 1989 Mar 23, 338(6213), 337 - 40
Transcripts of one of two Drosophila cyclin genes become localized in pole cells during embryogenesis; Whitfield WG et al.; Cyclins, originally discovered in the eggs of marine invertebrates, are proteins which undergo dramatic cycles of synthesis followed by degradation at the metaphase-anaphase transition of cell division . That they participate in the G2-M transition is supported by the fact that when synthetic cyclin messenger RNAs from clam and sea urchin are microinjected into the G2-arrested oocytes of Xenopus, they induce maturation . The cyclin of fission yeast is the product of the cdc13 gene, which is known to interact with cdc2, a gene required for the entry into mitosis . We have cloned the genes that encode A-type and B-type cyclins from Drosophila melanogaster by virtue of their sequence similarity to oligonucleotides corresponding to conserved regions of the cyclin genes . We show that both genes encode abundant maternal mRNAs, but whereas the cyclin A mRNA is relatively uniformly distributed before cell formation, the cyclin B mRNA becomes localized to the developing pole cells . In larvae, cyclin A is expressed predominantly in brain and imaginal disks, whereas cyclin B transcripts are abundant in testes.

J Biol Chem, 1989 Mar 15, 264(8), 4571 - 6
Deduced protein sequence of bone small proteoglycan I (biglycan) shows homology with proteoglycan II (decorin) and several nonconnective tissue proteins in a variety of species; Fisher LW et al.; The small proteoglycans (PG) of bone consist of two different molecular species: one containing one chondroitin sulfate chain (PG II) and the other, two chains (PG I) . These two proteoglycans are found in many connective tissues and have Mr = 45,000 core proteins with clear differences in their NH2-terminal sequences . Using antisera produced against synthetic peptides derived from the human PG I and PG II NH2 termini, we have isolated several cDNA clones from a lambda gt11 expression library made against mRNA isolated from human bone-derived cells . The clones, which reacted with antisera to the PG II peptide, were sequenced and found to be identical with the PG II class of proteoglycan from human fibroblasts known as PG-40 or decorin . The clones reacting to the PG I antisera, however, had a unique sequence . The derived protein sequence of PG I showed sufficient homology with the PG II sequence (55% of the amino acids are identical, with most others involving chemically similar amino acid substitutions) to strongly suggest that the two proteins were the result of a gene duplication . PG II (decorin) contains one attached glycosaminoglycan chain, while PG I probably contains two chains . For this reason, we suggest that PG I be called biglycan . The biglycan protein sequence contains 368 residues (Mr = 42,510 for the complete sequence and Mr = 37,983 for the secreted form) that appears to consist predominantly of a series of 12 tandem repeats of 24 residues . The repeats are recognized by their conserved leucines (and leucine-like amino acids) in positions previously reported for a diverse collection of proteins (none of which is thought to be proteoglycans) including: two morphogenic proteins (toll and chaoptin) in the fruit fly; a yeast adenylate cyclase; and two human proteins, the von Willebrand Factor-binding platelet membrane protein, GPIb, and a rare serum protein, leucine-rich glycoprotein.

Biochem J, 1989 Mar 15, 258(3), 817 - 21
Inhibition of mitochondrial-matrix inorganic pyrophosphatase by physiological {Ca2+}, and its role in the hormonal regulation of mitochondrial matrix volume; Davidson AM et al.; 1 . The pyrophosphatase activity in cytosolic and mitochondrial fractions of rat liver was 1.7 and 0.26 units/mg of protein respectively when assayed at 37 degrees C in the presence of physiological {Mg2+} (0.3 mM) . 2 . Approx . 80% of the mitochondrial pyrophosphatase was inaccessible to extramitochondrial PPi, of which 40% represented soluble matrix enzyme (0.38 unit/mg of matrix protein) . 3 . Ca2+ inhibited the soluble matrix enzyme; the effective K0.5 for inhibition increased as {Mg2+}, an essential cofactor of the enzyme, increased . Measured values were 0.39, 1.15, 3.7, 8.3 and 12.5 microM at 0.04 mM-, 0.1 mM-, 0.3 mM-, 0.6 mM- and 1 mM-Mg2+ respectively . 4 . The data were analysed by a kinetic model similar to that for yeast pyrophosphatase, which assumes the substrate to be MgPPi (Km 5 microM) with Mg2+ also activating at an additional site (K0.5 23 microM) . Ca2+ inhibits through the formation of CaPPi, a strong competitive inhibitor (Ki 0.067 microM) . 5 . Heart mitochondria also contain a soluble matrix pyrophosphatase of similar activity to that of liver mitochondria and with the same sensitivity to {Ca2+} . 6 . The data provide an explanation for the increase in mitochondrial PPi, mediated by Ca2+, which is responsible for the increase in matrix volume induced by gluconeogenic hormones {Davidson & Halestrap (1988) Biochem . J . 254, 379-384}.

Nucleic Acids Res, 1989 Mar 11, 17(5), 2023 - 42
The mitochondrial plasmid from Neurospora intermedia strain Labelle-1b contains a long open reading frame with blocks of amino acids characteristic of reverse transcriptases and related proteins; Pande S et al.; We have determined the DNA sequence of the 4070 base pair mitochondrial plasmid from the Labelle-1b strain of Neurospora intermedia . Analysis of the sequence revealed that the plasmid contains a long open reading frame (ORF) that could encode a protein of up to 1151 amino acids . Codon usage in the long ORF shows no clear relationship to Neurospora mitochondrial genes, nuclear genes, nor to the ORF of a different Neurospora mitochondrial plasmid . The long ORF contains regions of similarity to yeast mitochondrial RNA polymerase as well as blocks of amino acids that are characteristic of reverse transcriptases and the ORFs of certain group II mtDNA introns (Michel and Lang, (1985) Nature 316,641) . The plasmid gives rise to specific transcripts, some of which may be unit length, and which carry the information for expression of the long ORF . The genetic organization and content of the plasmid suggest that it is related to mobile genetic elements.

Nature, 1989 Mar 9, 338(6211), 170 - 2
DNA-protein conjugates can enter mitochondria via the protein import pathway; Vestweber D et al.; Mitochondria import most of their proteins and small molecules from the cytoplasm . There is some tentative evidence that they import some of their RNAs, but it is not known how nucleic acids could enter mitochondria . Here, we show that isolated yeast mitochondria can import a single-stranded or double-stranded 24-base pair piece of DNA whose 5' end is covalently linked to the C-terminus of a mitochondrial precursor protein.

Biochem Pharmacol, 1989 Mar 1, 38(5), 773 - 9
Ability of 1-methyltetrazole-5-thiol with microsomal activation to inhibit aldehyde dehydrogenase; Lipsky JJ; Antibiotics that contain the 1-methyltetrazole-5-thiol (MTT) leaving group are associated with an adverse effect when alcohol is ingested after their administration . Therefore, the ability of MTT to inhibit an enzyme in alcohol metabolism, aldehyde dehydrogenase (ALDH), was examined . In the absence of microsomes, MTT did not inhibit ALDH obtained from either yeast or rat liver . In the presence of rat hepatic microsomes, MTT was able to inhibit the enzyme from both sources . The characteristics of the inhibition were studied, using the yeast enzyme, and found to be dependent upon the length of incubation with the hepatic microsomes and upon the concentration of MTT . Inhibition required the presence of NADH and was not detected if the microsomes were heat treated . Dilution did not reverse the inhibition . Intact antibiotics which contain the MTT moiety did not cause an inhibition of yeast ALDH unless the antibiotics were first treated with potassium hydroxide and then incubated with microsomes . Inhibition of ALDH activity measured in the mitochondrial plus microsomal fractions of rat liver also required NADH and was prevented by glutathione and heat treatment of the microsomes . These results indicate that microsomal activation of MTT is necessary for inhibition of aldehyde dehydrogenase . The behavior of MTT described here may explain the adverse effect observed if alcohol is ingested following administration of MTT-containing antibiotics.

Biochim Biophys Acta, 1989 Mar 1, 1007(2), 176 - 83
Studies of interactions of porphyrins with transfer RNA by high-resolution NMR; Birdsall WJ et al.; The interactions of tetra-4N-methylpyridyl porphyrin and its zinc(II), copper(II) and manganese(III) complexes with brewer's yeast type V phenylalanine specific tRNA have been evaluated by high-resolution NMR . Differences in chemical shifts have been noted for three proton resonances in response to the presence of small quantities of the free base and the zinc and copper complexes . The protons giving rise to these signals are located on bases T54 and psi 55, both of which are involved in the primary intraloop and interloop hydrogen bonds that hold the D and T psi C loops together in the tertiary structure . In addition, broadening of specific resonances due to hydrogen bonding protons in the D stem at low ratios of porphyrin to tRNA indicates that the association of porphyrins increases the rate of imino proton exchange . The titration of the tRNA with the manganese(III) complex did not reveal shifts or specific broadening comparable to the other porphyrins at low ratios . The changes induced in the NMR spectrum of tRNA by porphyrins define their site of interaction with the polynucleotide . This site, at the outside of the elbow-bend in the tRNA 'L', is different from the locus of binding in tRNA for other classical DNA intercalators . Furthermore, a new mode of binding may be involved that is neither intercalative nor simply electrostatic.

J Invest Dermatol, 1989 Mar, 92(3), 385 - 90
A novel histone-stimulated protein kinase in normal and psoriatic epidermis; Takematsu H et al.; During the course of studies on protein kinases in psoriatic epidermis, a novel histone-activated protein kinase activity was identified . This activity (referred to as PK-II because it was the second peak of protein kinase activity eluted from a DEAE column) was partially purified from the supernatant of an epidermal homogenate by DEAE-cellulose column chromatography . Although histone was not a substrate for phosphorylation, in the presence of histone, endogenous proteins of Mr 105 and 95 kDa were phosphorylated . Activity was not affected by Ca2+/phospholipid, cAMP, cGMP, cAMP-dependent kinase inhibitor, spermine, spermidine, calmodulin, EGF, or phorbol ester . Phosphorylation was specific for serine and threonine residues . A major peak of PK-II activity eluted from sepharose 6B with an apparent Mr of 100 kDa, suggesting that histone may stimulate autophosphorylation . The properties of PK-II resemble those recently described for a class of polypeptide-dependent protein kinases isolated from placenta, Ehrlich ascites tumor cells, and bakers' yeast . PK-II was significantly higher in psoriatic involved epidermis (32.6 +/- 11.6 pmol/min/mg protein) compared to psoriatic uninvolved epidermis (5.7 +/- 0.6 pmol/min/mg; p = 0.03) and normal epidermis (9.5 +/- 2.2 pmol/min/mg; p = 0.05) . The function of histone stimulated protein kinase in epidermal function and its role in the pathophysiology of psoriasis remain to be explored.

Chem Pharm Bull (Tokyo), 1989 Mar, 37(3), 775 - 7
Pharmacological studies on EB-382, a new non-steroidal antiinflammatory agents: mode of action of the analgesic effects; Fujiyoshi T et al.; The analgesic mechanism of (+/-)-2-{p-{(2-methylallyl)amino}phenyl}propionic acid (EB-382), a new non-steroidal antiinflammatory agent, was studied . EB-382 exerted a potent analgesic effect on yeast-induced hyperalgesia in rats and bradykinin-induced writhing in mice, and its potency was superior to that of ibuprofen . EB-382 had a comparatively weak inhibitory effect on the prostaglandin E2 production from arachidonic acid in sheep seminal vesicle microsomal fraction in vitro . EB-382 exhibited a dose-dependent inhibitory effect on the phospholipase A2 activity in 3T3 mouse fibroblast cells with a different mechanism from steroidal agents, although indomethacin and ibuprofen did not reveal any significant inhibition . EB-382 did not affect the bradykinin-induced contraction of isolated rat uterus . Intracisternal injection of EB-382 did not affect the acetic acid-induced writhing response in mice . From these results, it is suggested that the analgesic effect of EB-382 in inflammation is exerted through its direct peripheral action, and inhibition of prostaglandin biosynthesis via phospholipase A2, rather than cyclooxygenase.

Z Naturforsch {C}, 1989 Mar-Apr, 44(3-4), 296 - 306
On the probability of a common origin for tRNA and 5S rRNA; Staves MP et al.; As part of a continuing study on the origin of genetic coding and the process of protein synthesis, we have compared sequences of a large number of transfer RNAs and several 5S ribosomal RNAs using automated routines . Transfer RNAs were found to exhibit a high degree of matching with 5S rRNAs . These matches are considered to be indicative of sequence homology, reflecting common ancestry of the two molecules . Other possible explanations for the matches (convergence, transfection) are discussed and found to be highly implausible . Matches are also found between 5S rRNA and the introns of yeast precursor tRNAs . Many matches extend from before the 3' end to after the 5' end of circularized 5S rRNA sequences . Data is presented which indicates a tandemly duplicated or circular molecule could have served as the precursor to both 5S rRNA and tRNA . A derivative of this molecule may have functioned as a universal translator before the evolution of the highly specialized tRNAs.

Placenta, 1989 Mar-Apr, 10(2), 125 - 35
The uterine lumen of the pregnant guinea-pig contains a large macrophage population; Sype W et al.; Cellular constituents of the uterine lumen were investigated . Fourteen pregnant sows of 40 + days' gestation were anaesthetized and naturally occurring peritoneal fluid was collected . A uterine horn was delivered and 0.25 ml Gey's solution injected into the uterine lumen to collect free cells . The fluid was aspirated into a syringe and the cells extracted, counted and prepared for phagocytosis experiments and microscopy . The cells were stained with alpha-naphthyl-acetate-esterase (ANAE) to determine the fraction that was non-specific esterase-positive, a feature of mononuclear phagocytes . Differential cells counts were also made . Both uterine and peritoneal compartments yielded large numbers of cells (greater than 10(6)/ml) . Peritoneal fluid cells were 47 +/- 6 per cent (SD) macrophages and 49 +/- 6 per cent eosinophils (the remainder being 'other' cells); 47 +/- 6 per cent also stained positively for ANAE . Uterine cells were 78 +/- 12 per cent macrophages, the remainder being mostly lymphocytes (18 +/- 11 per cent); 85 +/- 13 per cent stained positively with ANAE . Electron microscopy of the uterine cells confirmed that most had morphology consistent with being mononuclear phagocytes . Uterine and peritoneal cells phagocytized carbon particles and yeast cells when incubated at 37 degrees C . The origin and role of this macrophage population is unknown but uterine lumenal macrophages may be present to remove antigen-antibody complexes thus facilitating uptake of maternally derived IgG by the fetal yolk sac.

J Gen Virol, 1989 Mar, 70 ( Pt 3), 513 - 24
The complete nucleotide sequence of plum pox virus RNA; Maiss E et al.; The complete nucleotide sequence of the RNA of an aphid non-transmissible plum pox virus (PPV-NAT) isolate has been determined from five overlapping cDNA clones . cDNA prepared by primer extension was used to determine the 5' terminus . The assembled RNA is 9741 nucleotides in length, excluding a 3' terminal poly(A) sequence . One large open reading frame starts at nucleotide positions 36 to 38 and is terminated with an UAG codon at positions 9522 to 9524 . The putative start codon is located at positions 147 to 149 . The encoded polyprotein has a predicted Mr of 353.8K . Comparison of cistrons from tobacco vein mottling virus and tobacco etch virus with those predicted for PPV-NAT indicated a similar genome organization . A highly conserved sequence of 12 nucleotides was found in the 5' non-coding region of these three potyviruses . The potential polyadenylation signal from yeast (UAUGU) was found in the 3' non-coding region of PPV-NAT and several other members of the potyvirus group.

Mol Cell Biol, 1989 Mar, 9(3), 1212 - 23
Isolation and sequence of four small nuclear U RNA genes of Trypanosoma brucei subsp . brucei: identification of the U2, U4, and U6 RNA analogs; Mottram J et al.; Trypanosomes use trans splicing to place a common 39-nucleotide spliced-leader sequence on the 5' ends of all of their mRNAs . To identify likely participants in this reaction, we used antiserum directed against the characteristic U RNA 2,2,7-trimethylguanosine (TMG) cap to immunoprecipitate six candidate U RNAs from total trypanosome RNA . Genomic Southern analysis using oligonucleotide probes constructed from partial RNA sequence indicated that the four largest RNAs (A through D) are encoded by single-copy genes that are not closely linked to one another . We have cloned and sequenced these genes, mapped the 5' ends of the encoded RNAs, and identified three of the RNAs as the trypanosome U2, U4, and U6 analogs by virtue of their sequences and structural homologies with the corresponding metazoan U RNAs . The fourth RNA, RNA B (144 nucleotides), was not sufficiently similar to known U RNAs to allow us to propose an identify . Surprisingly, none of these U RNAs contained the consensus Sm antigen-binding site, a feature totally conserved among several classes of U RNAs, including U2 and U4 . Similarly, the sequence of the U2 RNA region shown to be involved in pre-mRNA branchpoint recognition in yeast, and exactly conserved in metazoan U2 RNAs, was totally divergent in trypanosomes . Like all other U6 RNAs, trypanosome U6 did not contain a TMG cap and was immunoprecipitated from deproteinized RNA by anti-TMG antibody because of its association with the TMG-capped U4 RNA . These two RNAs contained extensive regions of sequence complementarity which phylogenetically support the secondary-structure model proposed by D . A . Brow and C . Guthrie (Nature {London} 334:213-218, 1988) for the organization of the analogous yeast U4-U6 complex.

Fiziol Zh, 1989 Mar-Apr, 35(2), 69 - 74
{The role of riboflavin and pantothenic acid in the reproductive function of swine}; Pocherniaeva GM; Results of the experiment have shown that in order to increase multiple pregnancy and quality of piglets the ration of sows during pubescence and in the reproductive period must contain not less than 5-6 mg of riboflavin and 20-22 mg of pantothenic acid per one feed unit . That level can be ensured either by addition of synthetic preparation of riboflavin (4-4.5 mg) and calcium d-pantothenate (10-15 mg), or by inclusion of about 5% of the fodder yeast mass into complete-ration combined feed.

Eur J Biochem, 1989 Mar 1, 180(1), 61 - 6
Correlation between enzyme activity and hinge-bending domain displacement in 3-phosphoglycerate kinase; Sinev MA et al.; Diffuse X-ray-scattering data give evidence for large-scale structural change in pig muscle 3-phosphoglycerate kinase upon substrate binding . Simultaneous binding of 3-phosphoglycerate and MgATP either to the unmodified enzyme or to its active methylated derivative leads to about an 0.1-nm decrease in radius of gyration . These data coincide well with the previous data for yeast 3-phosphoglycerate kinase . When, instead of methylation, the two reactive thiol groups of pig muscle 3-phosphoglycerate kinase are carboxamidomethylated, the enzyme becomes inactive and the radii of gyration of its 'apo' and 'holo' forms do not differ within limits of experimental error . Thus, a correlation exists between the activity of 3-phosphoglycerate kinase and its substrate-induced large-scale conformational change . This correlation is a strong argument in favor of the functional importance of domain locking in the reaction catalyzed by 3-phosphoglycerate kinase.

Eur J Biochem, 1989 Mar 1, 180(1), 191 - 7
The primary structure of alcohol dehydrogenase from Drosophila lebanonensis . Extensive variation within insect 'short-chain' alcohol dehydrogenase lacking zinc; Villarroya A et al.; Insect alcohol dehydrogenase is highly different from the well-known yeast and mammalian alcohol dehydrogenases . The enzyme from Drosophila lebanonensis has now been characterized by protein analysis and was found to have a 254-residue protein chain with an acetyl-blocked N-terminal Met . Comparisons with the structures of the enzyme from other species allows judgement of the extent of variability within the insect alcohol dehydrogenases . They have diverged to a considerable extent; two forms analyzed at the protein level differ at 18% of all residues, and all the known Drosophila alcohol dehydrogenase structures reveal differences at 72 positions . Some deviations, against a background similarity, in the extent of changes are noted among the parts corresponding to different exons . The structural variation within Drosophila is about as large as the one for the mammalian zinc-containing alcohol dehydrogenase . Consequently, the results illustrate Drosophila relationships and establish great variations also for group of alcohol dehydrogenases lacking zinc.

Biochem J, 1989 Mar 1, 258(2), 535 - 40
Alterations in the protein-synthesis, -degradation and/or -secretion rates in hepatic subcellular fractions of selenium-deficient mice; Otter R et al.; Single mouse livers were subfractionated by differential centrifugation and isopycnic centrifugation on sucrose or metrizamide gradients and separated into subcellular compartments . The fractionation procedure was highly reproducible and yielded essentially similar results in different preparations of livers from selenium-adequate (Se+) and selenium-deficient (Se-) mice that were fed on a Torula-yeast-based diet containing less than 10 parts per 10(9) of selenium for at least 16 weeks . Mice of both dietary groups were injected intraperitoneally with 370 kBq of L-{U-14C}leucine, and 48 h later 1.85 MBq of L-{4,5-3H}leucine was injected intraportally . After another 1 h, the livers were removed and subjected to subcellular fractionation . Incorporation of the 3H label into proteins of the subcellular fractions was taken as a measure of relative protein-synthesis rate . The ratio of the 3H to the 14C protein-bound label of the same fractions was used as an estimate of relative protein-degradation and/or -secretion rate . The results showed a statistically significant 180% increase in protein-synthesis rate in the endoplasmic reticulum and a 80% increase in relative protein-degradation and/or -secretion rate in the same compartment . A significant decrease in the 3H/14C ratio, by 40 and 30% respectively, was observed in the Golgi fraction and in liver homogenate . The observed changes suggest a highly regulated hepatic response to selenium deficiency.

Clin Exp Immunol, 1989 Mar, 75(3), 466 - 70
Evaluation of phagolysosome fusion in acridine orange stained macrophages infected with Histoplasma capsulatum; Taylor ML et al.; A phagosome-lysosome (PL) fusion was performed in vitro using peritoneal cells from normal BALB/c mice and the J774.2 macrophage cell line infected with the yeast phase of the fungus Histoplasma capsulatum at ratios of 5 x 10(5), 5 x 10(6) or 1 x 10(7) yeasts per 1 x 10(6) macrophages, and phagocytosis was allowed to proceed for 5, 30 and 60 min . Macrophage lysosomes were pre-labelled with acridine orange and the cells were challenged with the parasite . Fusion was evaluated by fluorescence microscopy and the number of macrophages with stained yeast cells was scored . The phagolysosome fusion frequency (PLFF) was calculated by subtracting the specific fusions of infected macrophages from the non-specific fusions of uninfected macrophages and normalizing the total number of bound yeasts . The PLFF was determined using different doses and strains of H . capsulatum . Results showed that PLFF in infected macrophages depends on the infection dose . Inhibition of PL fusion was detected mainly at a high infection ratio (1 x 10(7) yeasts/1 x 10(6) macrophages), and PL fusion varied with phagocytosis time . No significant differences were observed in the fusions when different Histoplasma strains were used . Results with J774.2 cells were similar to peritoneal cells, indicating that both methodology and fusion calculations employed were useful, in spite of the heterogeneity of macrophage populations used.

Genetics, 1989 Mar, 121(3), 501 - 16
Molecular genetics of the Caenorhabditis elegans heterochronic gene lin-14; Ruvkun G et al.; We describe a general strategy for the genetic mapping in parallel of multiple restriction fragment length polymorphism (RFLP) loci . This approach allows the systematic identification for cloning of physical genetic loci within about 100 kb of any gene in Caenorhabditis elegans . We have used this strategy of parallel RFLP mapping to clone the heterochronic gene lin-14, which controls the timing and sequence of many C . elegans postembryonic developmental events . We found that of about 400 polymorphic loci in the C . elegans genome associated with the Tc1 family of repetitive elements, six are within 0.3 map unit of lin-14 . The three closest lin-14-linked Tc1-containing restriction fragments were cloned and used to identify by hybridization an 830-kb region of contiguous cloned DNA fragments assembled from cosmid and yeast artificial chromosome libraries . A lin-14 intragenic recombinant that separated a previously cryptic lin-14 semidominant mutation from a cis-acting lin-14 suppressor mutation was used to map the location of the lin-14 gene to a 25-kb region of this 830-kb contig . DNA probes from this region detected lin-14 allele-specific DNA alterations and a lin-14 mRNA . Two lin-14 semi-dominant alleles, which cause temporally inappropriate lin-14 gene activity and lead to the reiterated expression of specific early developmental events, were shown to delete sequences from the lin-14 gene and mRNA . These deletions may define cis-acting sequences responsible for the temporal regulation of lin-14.

Mol Biochem Parasitol, 1989 Mar 1, 33(2), 123 - 34
Perturbation of sterol biosynthesis by itraconazole and ketoconazole in Leishmania mexicana mexicana infected macrophages; Hart DT et al.; The azole antifungals ketoconazole and itraconazole possess in vitro antileishmanial activity against Leishmania mexicana mexicana amastigotes in macrophages (cell line J774G8) . As in yeast and fungi, the activity is likely to be due to inhibition of the cytochrome P-450-dependent 14 alpha-demethylation of lanosterol and/or 24,25-dihydrolanosterol . Indeed, 50% inhibition of ergosterol synthesis was observed at 0.21 microM ketoconazole and 0.15 microM itraconazole . At 5 microM ketoconazole, traces of ergosterol could be found, whereas no ergosterol could be detected in cells treated with 5 microM itraconazole . The inhibition of ergosterol biosynthesis was concomitant with an accumulation of the 14 alpha-methylsterols lanosterol and 24,25-dihydrolanosterol . Fifty percent inhibition of cholesterol synthesis in uninfected macrophages was achieved at 0.95 microM and 1.5 microM itraconazole and ketoconazole, respectively . In infected macrophages all {14C}acetate was incorporated in ergosterol, suggesting an inhibition in cholesterol synthesis in the host cells . An inhibition of ergosterol synthesis coincided with increasing cholesterol synthesis . The latter synthesis was inhibited at concentrations greater than 1 microM . However, even at 5 microM cholesterol synthesis was higher than under control conditions.

Gan To Kagaku Ryoho, 1989 Mar, 16(3 Pt 2), 522 - 9
{Natural UAG suppressor glutamine tRNA in retrovirus infected cells}; Kuchino Y; Mammalian cells contain two species of glutamine tRNAs, tRNA(CUGGIn) and tRNA(UmUGGIn) . The later minor glutamine tRNA which has the UmUG anticodon sequence can recognize an UAG amber termination codon of natural mRNA in an in vitro translation system . Recognition of the UAG nonsense codon by mammalian tRNA(UmUGGIn) is facilitated by two wobble base-pairs at the first and third position of the anticodon . Such unorthodox interaction between the codon and the anticodon which is not in accordance with the wobble hypothesis or the two out of three reading mechanism has been shown only in the recognition of the UAG nonsense codon by natural suppressor tRNA such as yeast tRNA(SGIn) and bovine liver tRNA(CAGLeu) . Due to such unique interaction with mRNA, the suppressor activity of mammalian glutamine tRNA(UmUGGIn) is weaker than that of tobacco tRNA(G psi ATyr), which is known to be a natural UAG suppressor tRNA in plants . Retrovirus infection followed by vegetative growth causes the selective and remarkable increase of the amount of UAG suppressor glutamine tRNA(UmUGGIn) in the virus-infected cells . The increased amount of tRNA(UmUGGIn) seems to be important not only for the sufficient production of a viral UAG readthrough protein, but also for the efficient translation of viral mRNAs, since tRNA(UmUGGIn) should read as efficiently the CAA glutamine codon which frequently appears in the viral genome . The increased level of tRNA(UmUGGIn) in virus-infected cells might be due to specific transcription activation of the tRNA gene for tRNA(UmUGGIn) . The factor required for the transcription regulation of the suppressor tRNA gene, if it exists in virus infected cells, may not be the same as the factors TFIIIB, IIIC and IIID so far identified . If such a specific transcription factor exists, it would be interesting to characterize it and to elucidate the mechanism by which it is induced by infection with Mo-MuLV or HIV.

Infect Immun, 1989 Mar, 57(3), 754 - 63
Inhibition of Leishmania donovani promastigote internalization into murine macrophages by chemically defined parasite glycoconjugate ligands; Palatnik CB et al.; Leishmania donovani, the agent of human visceral leishmaniasis, is an intracellular parasite that must be recognized and internalized by host macrophages to complete its biological cycle . In a search for possible ligands for macrophage surface receptors, glycoconjugates were obtained from Leishmania promastigotes by aqueous, phenol-aqueous, and alkaline extraction . A fucose-mannose glycoproteic ligand, a lipopeptidephosphoglycan, and a phosphate mannogalactan ligand were purified from promastigotes and analyzed for their chemical contents, with special attention to their glycidic moieties . Sugars that were identified as components of these glycoconjugates were tested for their capacity to inhibit promastigote internalization by BALB/c peritoneal macrophages in vitro . Neutral hexoses showed little inhibitory activity; fucose, charged monosaccharides, and a mannose polymer showed the highest activity . Two of the glycoconjugates (fucose-mannose glycoproteic ligand and phosphate mannogalactan ligand) purified from promastigotes were potent inhibitors of internalization, 75% inhibition being obtained at concentrations of 6 to 10 micrograms/ml . The simultaneous presence of both ligands in low concentrations yielded an increase in inhibitory activity above that found for each ligand alone, indicating that promastigotes may use at least two receptor sites for penetration into macrophages . These ligands are specific inhibitors of L . donovani promastigote phagocytosis, since 10 micrograms of each ligand per ml interfered neither with internalization of yeast cells nor with phagocytosis of Leishmania adleri promastigotes.

Mol Cell Biol, 1989 Mar, 9(3), 1120 - 7
cys-3, the positive-acting sulfur regulatory gene of Neurospora crassa, encodes a protein with a putative leucine zipper DNA-binding element; Fu YH et al.; The sulfur-regulatory circuit of Neurospora crassa consists of a set of unlinked structural genes which encode sulfur-catabolic enzymes and two major regulatory genes which govern their expression . The positive-acting cys-3 regulatory gene is required to turn on the expression of the sulfur-related enzymes, whereas the other regulatory gene, scon, acts in a negative fashion to repress the synthesis of the same set of enzymes . Expression of the cys-3 regulatory gene was found to be controlled by scon and by sulfur availability . The nucleotide sequence of the cys-3 gene was determined and can be translated to yield a protein of molecular weight 25,892 which displays significant homology with the oncogene protein Fos, yeast GCN4 protein, and sea urchin histone H1 . Moreover, the putative cys-3 protein has a well-defined leucine zipper element plus an adjacent charged region which together may make up a DNA-binding site . A cys-3 mutant and a cys-3 temperature-sensitive mutant lead to substitutions of glutamine for basic amino acids within the charged region and thus may alter DNA-binding properties of the cys-3 protein.

J Pharmacobiodyn, 1989 Mar, 12(3), 159 - 63
Studies on pharmacological activation of human serum immunoglobulin G by chemical modification and active subfragments . VIII . Effect of carboxamide-methylated light chain (Fr.I-L) on leukocyte functions; Mimura T et al.; Human serum immunoglobulin G light chain (Fr.I-L), which was reduced and carboxamide-methylated, showed no effect on formyl-methionyl-leucyl-phenylalanine (FMLP)-induced chemotaxis nor on phagocytosis of yeasts when directly added to guinea pig polymorphonuclear leukocytes (PMNs) . However, intravenously administered Fr.I-L inhibited emigration of leukocytes into the peritoneal cavity and promoted phagocytosis of yeasts in a yeast-induced peritonitis model in mice . Moreover, Fr.I-L reduced FMLP-induced chemiluminescence (CL) from PMNs . These facts indicated that the anti-inflammatory action of Fr.I-L was caused by inhibiting emigration of leukocytes into the injured site and scavenging superoxide radicals from the cells.

Gan To Kagaku Ryoho, 1989 Mar, 16(3 Pt 2), 646 - 52
{Biochemical characterization of tumor rejection antigen (TRA) and genetic control in anti-TRA immune responses}; Fujiwara H et al.; A tumor antigen capable of inducing tumor resistance (tumor rejection antigen; TRA) was obtained in a solubilized form by sodium dodecyl sulfate (SDS)-extraction of plasma membrane fraction from Rous sarcoma virus (RSV)-induced CSA1M fibrosarcoma cells (BALB/c origin) . Analyses by Sephacryl S-300 gel filtration and SDS-polyacrylamide gel electrophoresis revealed that TRA activity was recovered in the fraction of an approximate molecular weight (m . w.) of 60kD . Such TRA was also found to exist in plasma membrane fraction from various RSV-induced tumor cells but not from tumor cells induced with agents other than RSV . Unfractionated crude SDS-solubilized preparation contained gp 70 as detected by rabbit anti-gp 70 antiserum, whereas this reactivity was lost in the fraction exhibiting an m.w . of about 60 kD . Since this fraction retained pp60src activity, the relation of TRA to pp60src was further investigated . pp60src was also obtained from the lysate of v-src-expressing yeast transformant . Immunization of BALB/c mice with such pp60v-src-containing lysate failed to induce any significant tumor protection . The above 60 kD fraction of CSA1M solubilized antigens was allowed to bind to Sepharose beads coupled with anti-pp60src monoclonal antibody and was depleted of pp60src . Immunization with the fraction depleted of pp60src activity (bead-unbound fraction) resulted in potent tumor protection . These results indicate that the CSA1M solubilized membranous component(s) which has an approximate m.w . of 60kD but is (are) distinct from functional pp60src functions as the TRA against RSV-induced CSA1M tumor cells . The results are discussed in the context of (1) the relationship between the src gene and the expression of TRA and (2) genetic control in immune responses to the TRA common to RSV-induced tumor cells.

Cell, 1989 Feb 24, 56(4), 549 - 61
Five intermediate complexes in transcription initiation by RNA polymerase II; Buratowski S et al.; A native gel electrophoresis DNA binding assay was used to resolve complexes formed on the adenovirus Major Late Promoter by general transcription factors and RNA polymerase II . Five sets of complexes containing distinct components were identified . These complexes were generated by sequential binding of TFIID, TFIIA, TFIIB, RNA polymerase II, and TFIIE . The relative positions of each of the factors in the complexes were determined by DNAase I footprint analysis . TFIIA, derived from yeast or mammalian cells, formed a complex with yeast TFIID and the TATA element . TFIIB bound to this complex and probably acts as a "bridge" to the polymerase and the initiation site . The addition of ATP or dATP, necessary for "activation" of transcription, resulted in an alteration of the footprint in the +20 to +30 region, the same area protected upon addition of TFIIE to the initiation complex . Addition of ribonucleotide triphosphates generated new complexes that contained accurately initiated transcripts associated with the transcription machinery and the template DNA . A model for the interactions of components in initiation of transcription by RNA polymerase II is proposed.

Arch Biochem Biophys, 1989 Feb 15, 269(1), 114 - 24
Inactivation of human arginine-143, lysine-143, and isoleucine-143 Cu,Zn superoxide dismutases by hydrogen peroxide: multiple mechanisms for inactivation; Horton PJ et al.; Site-specific mutants of human Cu,Zn superoxide dismutase (Cu,ZnSOD) have been prepared in which the active-site arginine at position 143 (i.e., SODR143) has been replaced by either lysine (SODK143) or isoleucine (SODI143) . As reported previously (W.F . Beyer, Jr., et al . (1987) J . Biol . Chem . 262, 11182-11187), SODK143 and SODI143 have 43 and 11%, respectively, of the catalytic activity of SODR143 . H2O2, at low concentrations, acts as an affinity reagent for the inactivation of SODR143 . At pH 9.0 and 25 degrees C, the process is characterized by a half-saturation constant for H2O2, K50, of 5.1 mM and a maximum pseudo-first-order rate constant for inactivation, Kmax, of 0.53 min-1 . At pH 11.5, the corresponding values are 0.63 mM and 1.23 min-1 . The active species in the inactivation is likely HO2-, as previously found with yeast and bovine Cu,ZnSODs (see C.L . Borders, Jr., and I . Fridovich (1985) Arch . Biochem . Biophys . 241, 472-476) . SODK143 is also inactivated by HO2- by an affinity mechanism, i.e., one where reversible binding of H2O2 (HO2-) is a prerequisite for inactivation . At pH values of 9.0 and 11.5, the kmax values are 0.92 and 1.08 min-1, respectively; however, the corresponding K50 values increase to 42.5 and 15.8 mM, respectively . SODI143 is also inactivated by H2O2, but no evidence for an affinity mechanism was found; instead, a second-order kinetic mechanism was observed . Inactivation of each of the three enzymes is accompanied by the loss of one histidine per subunit . At elevated concentrations of H2O2, a second nonaffinity mechanism of inactivation of both SODR143 and SODK143 was found, in which a second equivalent of H2O2 reacts with the Cu,ZnSOD.HO2- complex to give a competing second-order inactivation . It appears that the positive charge of arginine-143 plays a role in the binding of HO2- at the active site of human Cu,ZnSOD, and that replacement of the arginine by lysine gives an enzyme with a similar affinity mechanism of inactivation, but with a greatly reduced affinity for HO2- . However, replacement with isoleucine causes an entirely different mechanism of inactivation; this raises the possibility that the mechanism of enzyme catalysis of superoxide dismutation by SODI143 is also different.

Biochim Biophys Acta, 1989 Feb 6, 1001(2), 196 - 200
Role of the 8-double bond of lanosterol in the enzyme-substrate interaction of cytochrome P-450(14DM) (lanosterol 14 alpha-demethylase); Aoyama Y et al.; The role of the 8-double bond of lanosterol in the enzyme-substrate interaction of yeast cytochrome P-450(14DM) (lanosterol 14 alpha-demethylase) was studied by analyzing metabolism of 8-lanostene-3 beta,32-diol, 7-lanostene-3 beta,32-diol, 6-lanostene-3 beta,32-diol and lanostane-3 beta,32-diol by the cytochrome . 8-Lanostene-3 beta,32-diol was actively metabolized by cytochrome P-450(14DM) and converted to the 32-nor-14-unsaturated metabolite . 7-Lanostene-3 beta,32-diol was also metabolized by the cytochrome, but the rate of metabolism was low . However, the cytochrome failed to catalyze the conversion of 6-lanostene-3 beta,32-diol and lanostane-3 beta,32-diol to their 32-nor metabolites . Spectral analysis of the sterol-cytochrome complexes and kinetics of cytochrome P-450(14DM) reduction in the presence of the sterols indicated that 6-lanostene-3 beta,32-diol and lanostane-3 beta,32-diol could not interact with the substrate site of the cytochrome . These results revealed that the 8-double bond of lanosterol plays an important role in the enzyme-substrate interaction of cytochrome P-450(14DM).

Biochem J, 1989 Feb 1, 257(3), 651 - 6
Extremely rapid endocytosis mediated by the mannose receptor of sinusoidal endothelial rat liver cells; Magnusson S et al.; Isolated sinusoidal endothelial rat liver cells (EC) in suspension bound and internalized ovalbumin, a mannose-terminated glycoprotein, in a saturable manner . The binding and uptake were Ca2+-dependent and were effectively inhibited by alpha-methyl mannoside and yeast mannan, but not by galactose or asialoglycoproteins . This corresponds to the binding specificity described for the mannose receptor of macrophages and non-parenchymal liver cells . Binding studies indicated a surface pool of 20,000-25,000 mannose receptors per cell, with a dissociation constant of 6 x 10(-8) M . Uptake and degradation of ovalbumin by isolated EC were inhibited by weak bases and ionophores which inhibit acidification of endocytic vesicles and dissociation of receptor-ligand complexes . Cycloheximide had no effect on uptake or degradation . Degradation, but not uptake, was inhibited by leupeptin . We conclude that ovalbumin dissociates from the mannose receptors in the endosomal compartment and the receptors are recycled to the cell surface, while the ovalbumin is directed to the lysosomes for degradation . A fraction of the internalized ovalbumin was recycled intact to the cell surface and escaped degradation (retroendocytosis) . The rate of internalization of ovalbumin by isolated EC was very fast, with a Ke (endocytotic rate constant) of 4.12 min-1, which corresponds to a half-life of 10 s for the surface pool of receptor-ligand complexes . To our knowledge, this is the highest Ke reported for a receptor-mediated endocytosis system.

Proc Natl Acad Sci U S A, 1989 Feb, 86(3), 930 - 2
Species distribution of a phosphoprotein (parafusin) involved in exocytosis; Satir BH et al.; A cytosolic phosphoprotein that appears to function in membrane fusion during exocytosis of secretory products has previously been isolated from Paramecium tetraurelia . This phosphoprotein, parafusin, with Mr 63,000, is rapidly dephosphorylated via a Ca2+-dependent process when secretagogues induce exocytosis in competent cells . Dephosphorylation does not occur in exocytosis-incompetent cells . Polyclonal antibodies against purified parafusin have now been used to show that this protein is present in unicellular organisms and cells of metazoan groups of wide evolutionary divergence, such as yeast, insects, and mammals, including humans . These results suggest that parafusin was present early in the history of eukaryotes and that it is of functional importance in the general mechanism of exocytosis and membrane fusion.

Wei Sheng Wu Xue Bao, 1989 Feb, 29(1), 45 - 50
{Culture conditions for uricase formation of Candida utilis}; Liu JG et al.; A uricase producing strain, Candida utilis AS 2.117, was screened out and the uricase-forming conditions have been investigated . The results are obtained as follows: uric acid, xanthine and guanine were found to be effective inducers for enzyme formation . Corn steep liquor was essential for enzyme formation and cell growth . Sucrose, glucose, D-mannose and D-fructose were the suitable carbon sources for enzyme formation . To the medium containing corn steep liquor, addition of inorganic nitrogen sources was harmful to the enzyme formation, however, addition of organic nitrogen sources enhanced enzyme activity slightly . The suitable medium for uricase formation was consisted of sucrose 5%, corn steep liquor 3%, uric acid 0.1%, proteose-peptone 0.1%, biotin 0.05%, KCl 0.1%, NaCl 0.1% at pH 6.2 . When the yeast was grown in 250 ml conic flask containing 30 ml of the medium mentioned above on the rotary shaker (200 r/min) at 25 degrees C for 21 h, uricase with 0.6 u/ml activity was obtained.

Nippon Yakurigaku Zasshi, 1989 Feb, 93(2), 61 - 73
{Pharmacological studies of a non-steroidal analgesic and antipyretic drug of LFP83}; Kuriyama K et al.; Pharmacological properties of LFP83, a non-steroidal analgesic and antipyretic drug, were studied in mice, rats and rabbits . LFP83 is a prodrug of flurbiprofen (FP) which is its active major metabolite in vivo . In experimental models of acetic acid writhing, the Randall and Selitto method, arthritic pain, yeast fever, LPS fever, carrageenin edema and adjuvant arthritis, LFP83 (i.v.) showed remarkable analgesic, antipyretic and antiinflammatory activities; and it was more potent than ketoprofen (i.m.) and aspirin DL-lysine (i.v.) . The analgesic activity of LFP83 was equal to or more potent than that of pentazocine, and its duration was longer than those of aspirin DL-lysine and pentazocine . In addition, the analgesic potency of LFP83 was approximately the same or more potent than that of FP (p.o.), and the onset of this analgesic effect began earlier . On the other hand, the ulcerogenic activity of LFP83 on rat gastric mucosa was less than that of FP (p.o.) in both single and consecutive (7 days) administrations . The safety index (UD50/ED50) of LFP83 was three to ten-fold higher than that of FP (p.o.) . As mentioned above, LFP83 is a potent analgesic, antipyretic and antiinflammatory drug; and in comparison to oral FP, it has a more potent and immediate effect, weak gastric irritation and high safety index.

Proc Natl Acad Sci U S A, 1989 Feb, 86(4), 1168 - 72
v-cbl, an oncogene from a dual-recombinant murine retrovirus that induces early B-lineage lymphomas; Langdon WY et al.; Cas NS-1 is an acutely transforming murine retrovirus that induces pre-B and pro-B cell lymphomas . Molecular cloning showed it was generated from the ecotropic Cas-Br-M virus by sequential recombinations with endogenous retroviral sequences and a cellular oncogene . The oncogene sequence shows no homology with known oncogenes but some similarity to the yeast transcriptional activator GCN4 . A 100-kDa gag-cbl fusion protein, with no detectable kinase activity, is responsible for the cellular transformation . The cellular homologue of v-cbl, present in mouse and human DNA, is expressed in a range of hemopoietic lineages.

Nihon Kyobu Shikkan Gakkai Zasshi, 1989 Feb, 27(2), 220 - 4
{A case of primary pulmonary sporotrichosis}; Nakano H et al.; We report the first case of primary pulmonary sporotrichosis in Japan . A 53-year-old man was admitted to our hospital for further examination of the abnormal shadows on chest X-ray film . Six months before admission, he was admitted to another hospital because of alcoholic liver disease and diabetes mellitus . Since the initial chest film showed cavities with infiltration in the left upper lung field, he was treated with antituberculous drugs despite negative sputum cultures for mycobacterium . In spite of the medication, his chest X-ray film revealed another cavitary lesion, so he was referred to our hospital . He had been asymptomatic during this period . Chest X-ray on admission disclosed multiple cavities in the left upper lobe and a cavity in the right lower lobe . Repeated sputum specimens, bronchial washings and brushings for cytology and cultures were all negative . In an attempt to clarify the pathogen, percutaneous lung aspiration (PLA) was performed . The PLA sample yielded a positive culture of Sporothrix shenckii . After the diagnosis, S . schenckii was also cultured from sputa . A sporothrix skin test and yeast agglutination test for S . schenckii were positive . In the absence of a history for skin lesion, the patient was diagnosed as a primary pulmonary sporotrichosis . As iodide therapy was ineffective, he was started on a regimen of intravenous amphotericin B . However his renal function progressively deteriorated, so amphotericin B was discontinued . Now he receives miconazole intravenously and is still under careful observation . As far as we know, this is the first report of primary pulmonary sporotrichosis in Japan . The possibility of sporotrichosis should be considered in any cases of undiagnosed cavitary lung diseases.

Jpn J Pharmacol, 1989 Feb, 49(2), 275 - 84
Stimulation of phagocytic activity of leukocytes and macrophages by traxanox sodium in mice and rats; Hisadome M et al.; Phagocytosis of yeast particles by mouse peritoneal macrophages or rat peritoneal polymorphonuclear leukocytes was enhanced by traxanox sodium in vitro . Traxanox sodium (30 and 100 mg/kg, p.o.) enhanced phagocytosis of yeast particles by leukocytes and macrophages in vivo or ex vivo . On the other hand, prednisolone, isoproterenol and theophylline inhibited phagocytosis by leukocytes and macrophages under the same conditions . Traxanox sodium (30 mg/kg, p.o.) prevented the suppression of phagocytosis by the above drugs . Combined with theophylline, isoproterenol synergistically inhibited phagocytosis by leukocytes in vivo . Traxanox sodium (100 mg/kg, p.o.) administered alone had no influence on carbon clearance in normal mice . However, traxanox sodium (1-30 mg/kg, p.o.) prevented the suppression of carbon clearance by the treatment with carrageenan, but not by the treatment with ethyl palmitate . These results suggest that traxanox sodium stimulates the phagocytic activity of leukocytes or macrophages and prevents the drug-induced suppression of the phagocytic activity of these cells.

Mycoses, 1989 Feb, 32(2), 78 - 83
Physiological characteristics of environmental isolates of pathogenic dematiaceous fungi; Gugnani HC et al.; A total of 39 environmental isolates of pathogenic dematiaceous fungi (Fonsecaea pedrosoi: 14 isolates, Phialophora verrucosa: 6, Cladosporium carrionii: 9, Exophiala jeanselmei: 2, Ramichloridium subulatum: 6, Cladosporium tenuissimum: 1 and Phaeoisaria clematidis: 1) were evaluated for their various physiological characteristics including the ability to produce extracellular enzymes . Significant physiological characteristics included the ability of isolates of all the species except C . tenuissimum to decompose tyrosine, lyse human red blood cells, conversion of R . subulatum from filamentous to yeast form when cultured on Czapek-Dox agar and also on blood agar and the growth of C . tenuissimum on Sabouraud dextrose agar incorporating 3.6 M NaCl . Isolates of all the species except those of C . carrionii produced lipase, while only the isolates of P . verrucosa, E . jeanselmei and Ph . clematidis were positive for phospholipase.

J Clin Oncol, 1989 Feb, 7(2), 159 - 67
Hematopoietic responses in patients with advanced malignancy treated with recombinant human granulocyte-macrophage colony-stimulating factor; Herrmann F et al.; The in vivo effect of yeast-derived recombinant human granulocyte-macrophage colony-stimulating factor (rh GM-CSF) was investigated in 30 patients with advanced malignancy in a phase Ib trial . Patients were treated at four different dose levels (120 to 1,000 micrograms/m2/d) by either daily intravenous (IV) bolus injection or 24-hour continuous infusion . Administration of rh GM-CSF resulted in a broad spectrum of dose- and schedule-dependent hematopoietic effects . Sustained infusion of rh GM-CSF elicited a maximum 17-fold average peak increase of the total WBC count with mainly neutrophils, eosinophils, and monocytes accounting for this rise, and increases in bone marrow cellularity with a shift to immature myeloid elements . Elevation of lymphocytes, platelets, and reticulocytes was not induced . Within five days after discontinuation of treatment the leukocytosis had disappeared . Adverse reactions encountered with rh GM-CSF seen in 65% of the patients studied were never life-threatening and always rapidly reversible . They included mild myalgias, facial flushing, low-grade fever, headache, bone discomfort, nausea, dyspnea, and transient decline of platelet counts . These results suggest that rh GM-CSF can be safely administered at the doses and schedules used and that it can induce in vivo some of the biological effects reported in in vitro studies . Although no objective antitumour responses have been seen, the ability of rh GM-CSF to increase number and function of leukocytes in vivo may prevent neutropenia and infections when GM-CSF is added to cytotoxic cancer therapy.

Exp Hematol, 1989 Feb, 17(2), 96 - 101
Selective culture of primate marrow-derived macrophages in medium devoid of protein additives; Patel JM et al.; Viable cultures of bone marrow-derived macrophages (BMM phi) from a primate source, the baboon, were maintained for up to 4 weeks in culture in the absence of any exogenous protein in the medium . Baboon peripheral blood monocytes, spleen, lung, and liver M phi s or human BMM phi failed to survive for greater than 4 days . The protein-free BMM phi cultures were morphologically distinctive by virtue of the extremely dendritic appearance of the M phi s . In contrast baboon marrow cultured in the presence of fetal calf serum led to the overgrowth of fibroblastoid cells and in the presence of horse serum produced numbers of giant cells or polykaryocytes in addition to M phi s . The BMM phi were capable of nonimmune phagocytosis of yeast particles, expressed Ia antigen on their surfaces (59%), and were positive cytochemically for nonspecific (alpha-naphthyl acetate) esterase, oil red O, and tartrate resistant acid phosphatase . The addition of sera to established protein-free BMM phi cultures induced a rapid change of shape, viz., retraction of the dendritic processes and rounding up of the M phi s apparent within 10 min . This shape change was not induced by the addition of hemopoietic growth factors granulocyte-macrophage colony-stimulating factor (GM-CSF), granulocyte CSF (G-CSF), macrophage CSF (M-CSF), or interleukin 3 (IL-3), nor could it be inhibited by the calcium channel blocking agent Nifedipine . Low levels of M-CSF activity, assayed by the murine bone marrow proliferation assay, were detected in the supernatant.

J Biomol Struct Dyn, 1989 Feb, 6(4), 729 - 39
Conformational change of the L-shaped tRNA(Phe) molecule; Nagamatsu K; Fluorophore of proflavine was introduced onto the 3'-terminal ribose moiety of yeast tRNA(Phe) . The distance between the fluorophore and the fluorescent Y base in the anticodon of yeast tRNA(Phe) was measured by a singlet-singlet energy transfer . Conformational changes of tRNA(Phe) with binding of tRNA(2Glu), which has the anticodon UUC complementary to the anticodon GAA of tRNA(Phe), were investigated . The distance obtained at the ionic strength of 100 mM K+ and 10 mM Mg2+ is very close to the distance from x-ray diffraction, while the distance obtained in the presence of tRNA(2Glu) is significantly smaller . Further, using a fluorescent probe of 4-bromomethyl-7-methoxycoumarin introduced onto pseudouridine residue psi 55 in the T psi C loop of tRNA(Phe), Stern-Volmer quenching experiments for the probe with or without added tRNA(2Glu) were carried out . The results showed greater access of the probe to the quencher with added tRNA(2Glu) . These results suggest that both arms of the L-shaped tRNA structure tend to bend inside with binding of tRNA(2Glu) and some structural collapse occurs at the corner of the L-shaped structure.

Genes Dev, 1989 Feb, 3(2), 158 - 72
The Hox-1.3 homeo box protein is a sequence-specific DNA-binding phosphoprotein; Odenwald WF et al.; We report that the murine Hox-1.3 homeo domain protein is a nuclear phosphoprotein capable of binding to specific DNA sequences . DNase I protection of the Hox-1.3 gene promoter region with the Hox-1.3 protein identifies a binding site 144 bp upstream from the start of transcription . Both phosphorylated and nonphosphorylated forms bind DNA directly in a sequence-specific manner . Electrophoretic mobility shift assays were performed with a set of synthetic oligonucleotides representing either the DNase I-protected region of the Hox-1.3 gene or partially homologous sequences present in promoter regions of other characterized viral, yeast, and mammalian genes . From the results, we deduce a consensus binding motif of CPyPyNATTAT/GPy . Base substitutions in the core ATTA sequence severely reduce or abolish binding . In the SV40 enhancer, the Hox-1.3 binding motif overlaps both the octamer (Octa2) and the transactivator protein-1 (AP-1) binding sites . The Hox-1.3 binding motif also overlaps the nuclear factor III (NF-III) octamer motif in the adenovirus-2 origin of DNA replication . Overlap among DNA-binding sites suggests that regulation imparted by certain cis-elements may be integrated by these different factors.

Mycopathologia, 1989 Feb, 105(2), 75 - 8
Treatment of disseminated histoplasmosis in hamsters; Finquelievich JL et al.; A comparative study between itraconazole, ketoconazole and amphotericin B in the treatment of experimental histoplasmosis in hamsters was carried out . Seventy five animals were inoculated intracardiacally with the yeast-phase of Histoplasma capsulatum . They were divided in 5 groups: 1) treated with itraconazole by gavage (g) at a daily dose of 16 mg/kg; 2) treated with ketoconazole by (g) at a daily dose of 80 mg/kg; 3) treated with amphotericin B intraperitoneally (i.p.) at 6 mg/kg every other day; 4) control animals receiving distilled water i.p . and 5) control animals receiving P.E.G . 200 by (g) . All the treatments were started one week after the challenge inoculation and they were given for 21 days . The results were evaluated by autopsy of all the animals one week after the end of the treatments . The following determinations were taken into account: microscopic examinations of spleen, liver and lungs and cultures of the spleen with determination of colony forming units/g . All the antifungal drugs used in this study were able to cause negative microscopic examinations of the liver, spleen and lungs; but only amphotericin B produced culture negative results . Itraconazole and ketoconazole presented 66% and 86% of positive cultures respectively, nevertheless the C.F.U . were lower than those obtained in control groups . In these experimental conditions amphotericin B seems to be more active than the azolic compounds and itraconazole is slightly superior to ketoconazole at a lower dose.

J Clin Microbiol, 1989 Feb, 27(2), 337 - 8
Unexpected isolation of Bordetella pertussis from patients with acquired immunodeficiency syndrome; Ng VL et al.; We isolated Bordetella pertussis from three patients with acquired immunodeficiency syndrome who underwent diagnostic bronchoscopy for evaluation of respiratory symptoms . The B . pertussis isolates were recovered from medium (charcoal-yeast extract agar) formulated to enhance recovery of Legionella spp., and one of the isolates stained positively with antisera directed against Legionella spp.

Biochem Biophys Res Commun, 1989 Jan 31, 158(2), 610 - 6
Secondary structure and orientation of a chemically synthesized mitochondrial signal sequence in phospholipid bilayers; Goormaghtigh E et al.; A pre-sequence of 25 amino acids is required for import of yeast cytochrome oxidase subunit IV into mitochondria . Structure and orientation of the 25 amino acids synthesized peptide (p25) in a lipid bilayer were investigated by infrared attenuated total reflection spectroscopy . This method allowed to overcome the difficulties related to the optical turbidity due to the light scattering on membrane fragments which prevents the use of circular dichroism . We demonstrate here that incubation of the peptide with DOPC (dioleoylphosphatidylcholine) and DOPC-CL (dioleoylphosphatidylcholine - cardiolipin) liposomes is accompanied by an increase in alpha-helical content as compared to beta structure . Polarisation measurements indicate that the amphipathic helical segment is inserted parallel to the lipid acyl chains in cardiolipin containing liposomes.

Gene, 1989 Jan 30, 75(1), 73 - 83
A salivary gland-specific, maltase-like gene of the vector mosquito, Aedes aegypti; James AA et al.; Genomic and cDNA clones of a gene expressed specifically in the salivary glands of adult Aedes aegypti have been isolated and sequenced . This gene encodes an abundant mRNA that is transcribed throughout the male salivary gland but only in the cells of the proximal lateral lobes of the female gland . The deduced protein has many basic amino acids, several possible sites for N-glycosylation, and displays striking similarities with the products of a yeast maltase gene and three previously unidentified genes from Drosophila melanogaster . We propose the name 'Maltase-like I' (MalI) to designate this gene . The presumed function of this gene product is to assist the mosquito in its sugar-feeding capabilities . The mosquito and fruitfly genes have similar structural features 5' to the protein coding regions, indicating that these genes may share common control mechanisms.

Nature, 1989 Jan 26, 337(6205), 376 - 80
Tnt1, a mobile retroviral-like transposable element of tobacco isolated by plant cell genetics; Grandbastien MA et al.; Transposable elements can be identified by their ability to induce mutant alleles at new loci . The retrotransposon family is thought to transpose through an RNA intermediate and has many similarities to vertebrate proretroviruses . In plants, retrotransposons have been described in maize, Arabidopsis and wheat, and non-viral retroposons in maize . Most of these elements, however, have been found as non-mobile integrated units . Here, we report the isolation of the first tobacco (Nicotiana tabacum) transposable element, Tnt1, which seems to be the most complete mobile retrotransposon characterized in higher plants . Tnt1 has been isolated after its transposition into the nitrate reductase (NR) structural gene of tobacco, and transposition events have been detected through in vitro selection of spontaneous NR-deficient (NR-) mutant lines in cell cultures derived from tobacco mesophyll protoplasts . Tnt1 is 5,334 nucleotides long, contains two 610-base-pair-long terminal repeats and a single open reading frame of 3,984 nucleotides . Comparison of the Tnt1 open reading frame coding potential with those of the Drosophila melanogaster copia retrotransposon, yeast Ty retrotransposon, and vertebrate proretroviruses revealed that Tnt1 is closely related to copia and carries all the functions known to be required for autonomous transposition by reverse transcription.

N Z Med J, 1989 Jan 25, 102(860), 1 - 3
The safety and immunogenicity of a recombinant hepatitis B vaccine in neonates; Gunn TR et al.; A study to evaluate the safety and immunogenicity of a yeast derived recombinant DNA hepatitis B vaccine (Engerix-B) was conducted in healthy newborn infants born to low risk European mothers negative for hepatitis B surface antigen (HBsAg) . The vaccination schedule using 20 micrograms doses was administered intramuscularly at 0, 1 and 6 months . The seroconversion rate was 99% (90 of 91 infants) . The geometric mean titer of antibody to hepatitis B was 1259 mIU/mL one month after the third dose of vaccine . Possible side effects reported by the mothers were minor and uncommon . This vaccine is highly immunogenic and safe for use in infants.

Nature, 1989 Jan 19, 337(6204), 270 - 2
Induction of endogenous IFN-alpha and IFN-beta genes by a regulatory transcription factor, IRF-1; Fujita T et al.; Interferons (IFNs) have an important role in cell growth and differentiation . The most well-known function of IFNs is their antiviral activity; viral infections result in induction of the transcription of the IFN-alpha and IFN-beta genes . Recently we isolated the gene encoding a transcription factor, IRF-1, that may play a part in the induction of IFN genes . Interestingly, the IRF-1 gene itself is virus-inducible, suggesting the importance of de novo production of IRF-1 in IFN gene induction . Here we show that high-level expression of the cloned mouse IRF-1 gene in monkey COS cells results in the induction of endogenous IFN-alpha and IFN-beta genes without viral stimulation . Furthermore, we demonstrate the induction of these genes by an IRF-1/yeast GAL4 chimaeric transcription factor . This may be the first demonstration of the specific induction of silent chromosomal genes by transfection of a single transcription factor gene in mammalian cells.

Nucleic Acids Res, 1989 Jan 11, 17(1), 147 - 61
Cre-stimulated recombination at loxP-containing DNA sequences placed into the mammalian genome; Sauer B et al.; The cre gene of coliphage P1 encodes a 38 kDa protein which efficiently promotes both intra- and intermolecular recombination at specific 34 bp sites called loxP . To demonstrate that the Cre protein can promote DNA recombination at loxP sites resident on a mammalian chromosome, a mouse cell line was constructed containing two directly repeated loxP sites flanking a 2.5 kb yeast DNA fragment and inserted between the SV40 promoter and the neo structural gene to disrupt expression of the neo gene . Expression of the cre gene in this cell line results in excision of the intervening yeast DNA and thus permits sufficient expression of the neo gene to allow cell growth in high concentrations of G418 . Southern analysis indicated that Cre-mediated excision occurred at the loxP sites . In the absence of the cre gene such excisive events are quite rare . Cre-mediated recombination should thus be quite useful in effecting a variety of genomic rearrangements in eukaryotic cells.

J Biol Chem, 1989 Jan 5, 264(1), 363 - 9
The amino acid sequence of rat liver glucokinase deduced from cloned cDNA; Andreone TL et al.; Rat liver glucokinase (ATP:D-hexose 6-phosphotransferase, EC 2.7.1.1) was purified to homogeneity, cleaved, and subjected to amino acid sequence analysis . Forty-five percent of the protein sequence was obtained, and this information was used to design oligonucleotide probes to screen a rat liver cDNA library . A 1601-base pair cDNA (GK1) contained an open reading frame that encoded the amino acid sequences found in the peptides used to generate the oligonucleotide probes . A second cDNA was subsequently identified (GK.Z2), which is 2346 base pairs long and corresponds to nearly the entire glucokinase mRNA . Blot transfer analysis of hepatic RNA showed that glucokinase mRNA exists as a single species of about 2400 nucleotides . Four hours of insulin treatment of diabetic rats resulted in a 30-fold induction of this mRNA . GK.Z2 has a long open reading frame which, with the known partial peptide sequence, allowed us to deduce the primary structure of glucokinase . The enzyme is composed of 465 amino acids and has a mass of 51,924 daltons . Glucokinase has 53 and 33% amino acid sequence identities with the carboxyl-terminal domains of rat brain hexokinase I and yeast hexokinase, respectively . If conservative amino acid replacements are also considered, glucokinase is similar to these two enzymes at 75 and 63% of positions, respectively . The putative glucose- and ATP-binding domains of glucokinase were identified, and these regions appear to be highly conserved in the hexokinase family of enzymes.

J Mol Biol, 1989 Jan 5, 205(1), 201 - 28
Structure of the complex of Streptomyces griseus proteinase B and polypeptide chymotrypsin inhibitor-1 from Russet Burbank potato tubers at 2.1 A resolution; Greenblatt HM et al.; A low molecular weight protein inhibitor of serine proteinases from Russet Burbank potato tubers, polypeptide chymotrypsin inhibitor-1 (PCI-1), has been crystallized in complex with Streptomyces griseus proteinase B (SGPB) . The three-dimensional structure of the complex has been solved at 2.1 A resolution by the molecular replacement method and has been refined to a final R-factor (= sigma{{Fo{-{Fc{{/sigma{Fo{) of 0.142 (8.0 to 2.1 A resolution data) . The reactive site bond of PCI-1 (Leu38I to Asn39I) is intact in the complex, and there is no significant distortion of the peptide from planarity . The distance between the active site serine O gamma of SGPB and the carbonyl carbon of the scissile bond of PCI-1 is 2.8 A (1 A = 0.1 nm) . The inhibitor has little secondary structure, having a three-stranded antiparallel beta-sheet on the side opposite the reactive site and four beta-turns . PCI-1 has four disulphide bridges; these presumably take the place of extensive secondary structure in keeping the reactive site conformationally constrained . The pairing of the cystine residues, which had not been characterized chemically, is as follows: Cys3I to Cys40I, Cys6I to Cys24I, Cys7I to Cys36I, and Cys13I to Cys49I . The molecular structure of SGPB in the PCI-1 complex agrees closely with the structure of SGPB complexed with the third domain of the turkey ovomucoid inhibitor (OMTKY3) . A least-squares overlap of all atoms in SGPB gives a root-mean-square difference of 0.37 A . One of the loops of SGPB (Ser35 to Gly40) differs in conformation in the two complexes by more than 2.0 A root-mean-square for the main-chain atoms . Thr39 displays the largest differences with the carbonyl carbon atom deviating by 3.6 A . This conformational alternative is a result of the differences in the molecular structures of the P'4 residues following the reactive site bonds of the two inhibitors . This displacement avoids a close contact (1.3 A) between the carbonyl oxygen of Ser38 of SGPB and Pro42I C beta of PCI-1 . The solvent structure of the PCI-1-SGPB complex includes 179 waters, two sulphate or phosphate ions, and one calcium or potassium ion, which appears to play a role in crystal formation . The molecular structure of PCI-1 determined here has allowed the proposal of a model for the structure of a two-domain inhibitor from potatoes and tomatoes, inhibitor II.(ABSTRACT TRUNCATED AT 400 WORDS)

Eur J Biochem, 1989 Jan 2, 178(3), 779 - 87
Conserved epitopes on plant H1 histones recognized by monoclonal antibodies; Mazzolini L et al.; A series of monoclonal antibodies specific for distinct regions of H1 histone from the plant Nicotiana tabacum were obtained from fusion experiments with spleen cells of mice immunized with tobacco nuclear extracts . These monoclonal antibodies were characterized and the evolutionary conservation of the epitopes in higher plants and animals studied by immunoblotting and enzyme-linked immunosorbent assay (ELISA) . Whereas some epitopes appear restricted to the Solanaceae plant family, others are common to all higher eukaryotes tested and even detectable on nuclear proteins of yeast . ELISA experiments performed with isolated tobacco chromatin give some indications of the differential accessibility of the epitopes after interaction of H1 histone with the nucleosome.

J Invest Dermatol, 1989 Jan, 92(1), 117 - 9
A new model for growth and filament production of Pityrosporum ovale (orbiculare) on human stratum corneum in vitro; Faergemann J; A new model for the production of hyphae in Pityrosporum ovale in vitro is described . P . ovale was cultured on human stratum corneum pieces placed directly on a culture medium . The highest number of hyphae (24%) was seen after 6 d of incubation at 37 degrees C in a microaerophilic environment . There was a variation between the strains tested . This model opened possibilities to study the filamentous form of P . ovale in vitro . The effect of antimycotics and variation in antigens may be investigated and compared to the yeast form.

J Cancer Res Clin Oncol, 1989, 115(5), 445 - 8
Increased mutagenicity of N-nitrosodiethanolamine in human lymphocyte cultures after activation by alcohol dehydrogenase; Henn I et al.; The effect of alcohol dehydrogenase (ADH/NAD) from yeast and horse liver was tested on the induction of chromosomal mutations and sister chromatid exchanges (SCE) by N-nitrosodiethanolamine (NDELA) in human lymphocyte cultures . BrdUrd (27 micrograms) was added 24 h after starting the cultures to allow visualisation of SCE . ADH/NAD and NDELA were added 24 h later in different concentrations . No significantly higher level of numerical or structural chromosome aberrations was observed . However, the SCE frequency per cell was significantly increased by adding NAD (31.25 mumol and 62.5 mumol) . The exclusive addition of 220 units ADH from yeast as well as 1.8 units ADH from horse liver also raised the number of SCE highly significantly . The combination of NAD and ADH was more effective than each substances alone in the yeast but not in the horse liver system . NDELA in a range of 12.5-62.5 mumol, given to cultures with ADH/NAD from yeast, additionally increased the SCE frequencies in a dose-dependent way . Similar results were found in cultures containing ADH/NAD from horse liver and 6.25-31.25 mumol NDELA, but the total numbers of SCE were distinctly higher . These results indicate that NDELA is strongly activated by ADH from yeast but even more by ADH from horse liver.

Yao Xue Xue Bao, 1989, 24(2), 151 - 4
{The antiinflammatory and immunostimulating activities of S-4001--a polysaccharide isolated from Lei Wan (Polyporus mylitiae)}; Wang WJ et al.; Lei Wan, Poliporus mylittae Cook et Mass (Omphalia lapidescens Schraet) is a kind of fungus used in traditional Chinese medicine, as an anthelminthic . From Lei Wan, an active component designated as S-4001 had been isolated . Preliminary results indicate that S-4001 belongs to D, beta, 1-3 glucan with some 1-6 linkages . After administration of S-4001, significant antiinflammatory activity was found in various experimental animal models, including croton oil induced ear edema in mice and agar or yeast induced ankle swelling in rats . An inhibitory action on leucocyte migration induced by intraperitoneal injection of CMC in rats was also observed . The plasma content of corticosterone was significantly increased, but the content of ascorbic acid in the adrenals did not change in rats given S-4001 . Apart from these actions, S-4001 showed a number of immunostimulating actions such as increasing the clearance of Congo red from mice blood and potentiating the immunohemolysis reaction in 615 mice.

Boll Chim Farm, 1989 Jan, 128(1), 33 - 7
Antinociceptive properties of lysozyme fragments; Bruzzese T et al.; The in vitro digestion of hen egg white lysozyme with artificial gastric juice gave a complex mixture of peptides, from which a peptide corresponding to the aminoacid sequence 39-53 was isolated . Its further hydrolysis with artificial enteric juice gave two smaller fragments having the aminoacid sequence 39-45 and 46-53 respectively . These products, like lysozyme, showed antinociceptive activity in rats against foot hyperalgesia induced by a subplantar injection of brewer's yeast.

Vopr Med Khim, 1989 Jan-Feb, 35(1), 52 - 5
{Activity of amine oxidases from blood plasma and thrombocytes in sensitization and allergy}; Shleikin AG et al.; Activities of amine oxidase from blood plasma and of monoamine oxidase from thrombocytes were studied in persons sensitized to yeast fungus and in patients with allergic diseases . Activity of the blood plasma amine oxidase tended to increase in sensitization and to decrease in patients with allergic dermatoses . In this group of patients activity of the thrombocyte monoamine oxidase was higher as compared with that of healthy persons.

Ann Clin Biochem, 1989 Jan, 26 ( Pt 1), 83 - 8
The effect of supplementation with selenium and vitamin E in psoriasis; Fairris GM et al.; Since reduced concentrations of selenium in whole blood, plasma and white cells had previously been observed in psoriasis, 69 patients were supplemented daily with either 600 micrograms of selenium-enriched yeast, 600 micrograms of selenium-enriched yeast plus 600 IU of vitamin E or a placebo for 12 weeks . Before supplementation, the patients' mean concentrations of selenium in whole blood and plasma were reduced compared with those of matched healthy controls but their red cell glutathione peroxidase (GSH-Px) activity was normal . After 12 weeks supplementation the patients' mean whole blood, plasma and platelet selenium concentrations, platelet GSH-Px activity and plasma vitamin E concentration had risen significantly from the baseline values but their mean skin selenium concentration and red cell GSH-Px activity remained unchanged . The mean white cell selenium concentration rose only in the group receiving selenium alone . Neither supplementation regimen reduced the severity of psoriasis or produced side-effects . The increase in platelet GSH-Px activity suggests that the supplements were bioavailable and that the patients' selenium status may have been reduced prior to supplementation . The failure of the selenium content of the skin to increase may explain why the patients' psoriasis remained unchanged during supplementation.

Ann Biol Clin (Paris), 1989, 47(2), 85 - 90
{Antifungigram: interpretation and choice of a technic}; Vermorel O et al.; Identification and in vitro susceptibility testing of fungi is necessary mainly in immunocompromised patients . So, after a survey of the epidemiological results, the authors specified the indications of susceptibility tests; their advantages and difficulties and the lots of variations related to the fungus, drug, medium and technic . The authors compared the more recent methods from their experience: susceptibility to 6 antifungals (amphotericin B, flucytosine and 4 imidazoles) was estimated in 122 yeast strains isolated from deep specimens by 5 marketed technics . They got a good correlation between Mycototal and Antifongigramme Pasteur, but the two other technics (Mycodisk and Neosensitabs) frequently showed unreliable results.

EMBO J, 1989 Jan, 8(1), 301 - 8
Ribosome inactivation by ricin A chain: a sensitive method to assess the activity of wild-type and mutant polypeptides; May MJ et al.; When recombinant ricin A chain transcripts are translated in a rabbit reticulocyte lysate the ribosomes are rapidly inactivated as shown by their inability to support translation of yeast preproalpha factor or chicken lysozyme transcripts added subsequently . In contrast, ribosomes which have translated transcripts encoding non-toxic polypeptides such as ricin B chain, readily translate the second transcript under identical conditions . Ribosome inactivation is accompanied by a highly specific modification of 28S rRNA which occurs at the same position as the N-glycosidic cleavage of an adenine residue and which is thought to cause inactivation of the ribosomes . Protein synthesis by wheat germ ribosomes was not inhibited under the conditions which inhibit reticulocyte ribosomes confirming earlier observations that plant cytoplasmic ribosomes are much less sensitive to inhibition by ricin A chain than are mammalian ribosomes . Using the same assay we have shown that deleting an internal hexapeptide, which shares homology with hamster elongation factor-2, completely abolishes catalytic activity . Deleting a second pentapeptide conserved between ricin A chain and the ribosome-inactivating plant toxin trichosanthin, had no effect . Deleting the first nine residues from the N-terminus of A chain did not affect toxicity whereas deleting a further three residues inactivated the polypeptide . Point mutations which individually converted arginine 48 and arginine 56 of ricin A chain to alanine residues or which deleted arginine 56 were also without effect on the catalytic activity of the toxin.

Mol Gen Genet, 1989 Jan, 215(2), 349 - 54
An element with long terminal repeats and its variant arrangements in the genome of Lilium henryi; Sentry JW et al.; A 9.35 kbp element with long terminal direct repeats (LTRs) of 2.4 kbp has been characterized from the large genome of Lilium henryi . The organization of the element, named del, was examined in 20 fragments from a genomic DNA library constructed using phage EMBL3 . Five fragments apparently contained full 9.35 kbp elements while in 11 del was recovered in part . Two clones carried tandem arrangements of del, with single LTRs and internal sequences alternating, and one insert contained a solo LTR . Evidence suggests that these arrangements, which can arise by unequal crossing over, have a genomic rather than a cloning origin . Finally, one cloned fragment had a complex del arrangement yet to be fully defined . Limited sequencing of five LTRs (from a full del, a tandem arrangement and a solo LTR) indicates that the consensus termini are 5'TGT...ACA3', with del sequences flanked by a 5 bp tandem repeat . Thus del shares properties with retrotransposons such as the copia family of Drosophila and Ty of yeast . However with more than 13,000 copies per genome it seems del has been amplified more than is usual for such elements.

Zentralbl Mikrobiol, 1989, 144(4), 219 - 30
{Dimorphism in fungi--a gray zone for taxonomy?}; Kreisel H et al.; Dimorphic fungi can grow as mycelial phase and as yeast phase . The change of growth form is effected by an altered programme in gene expression, which is induced either in certain stages of ontogenesis or by environmental factors . Therefore it is necessary to distinguish ontogenetically conditioned (irreversible) and environment conditioned (reversible) dimorphism . The first is characteristic for dimorphic Ustomycetes, Basidiomycetes and related anamorphs as well as for Taphrinales; the second for the majority of dimorphic Ascomycetes, Endomycetes, and related anamorphs . The integration of dimorphic fungi in the systems of filamentous fungi and of yeasts--which originally have been elaborated rather independently--was difficult in many cases . In recent times, the study of certain ultrastructural and biochemical-physiological characters has much facilitated the taxonomic arrangement of dimorphic fungi and has allowed to demonstrate some phylogenetic relations among filamentous, dimorphic, and yeast fungi . The authors hold the concept that yeasts have evolved from filamentous resp . dimorphic fungi by neotenic simplification.

Cell Motil Cytoskeleton, 1989, 13(4), 245 - 63
Indirect immunofluorescence localization of ponticulin in motile cells; Wuestehube LJ et al.; Ponticulin is the major actin-binding integral glycoprotein in plasma membranes isolated from log-phase Dictyostelium discoideum amebae . As such, this protein appears to be an important link between the plasma membrane and actin filaments (Wuestehube and Luna: Journal of Cell Biology 105:1741-1751, 1987) . In this study, indirect immunofluorescence microscopy was used to examine the distribution of ponticulin in randomly moving D . discoideum amebae and in amebae engaged in cell migration and phagocytosis . Ponticulin is distributed throughout the plasma membrane and also is present in intracellular vesicles associated with the microtubule-organizing center-Golgi complex adjacent to the nucleus . In aggregating amebae, ponticulin is concentrated in regions of lateral cell-cell contact and in arched regions of the plasma membrane . Ponticulin also is present, but not obviously enriched, in filopodia, in the actin-rich anterior end of polarized cells, and in detergent-insoluble cytoskeletons . In amebae engaged in phagocytosis of yeast, ponticulin is present but not enriched in phagocytic cups and is associated with intracellular vesicles around engulfed yeast . These results suggest that ponticulin is stably associated with actin filaments in certain regions of the plasma membrane and that the actin-binding activity of ponticulin may be tightly controlled . Indirect immunofluorescence microscopy and immunoblot analysis demonstrate that human polymorphonuclear leukocytes also contain a 17 kD protein that specifically cross-reacts with antibodies affinity-purified against D . discoideum ponticulin . As in D . discoideum, the mammalian 17 kD ponticulin-analog appears to be localized in plasma membrane and is evident in actin-rich cell extensions . These results indicate that ponticulin-mediated linkages between the plasma membrane and actin may be present in higher eukaryotic cells.

J Clin Lab Anal, 1989, 3(4), 209 - 14
Competitive ELISA for detection of native ras gene-related products in sera of cancer patients; Epelbaum R et al.; A solid phase, enzyme-linked immunosorbent assay (ELISA) competition kit was developed to detect circulating native ras gene-related products in sera of 151 healthy volunteers and cancer patients . This assay uses monoclonal antibody (mAb) BST-6A generated against a yeast-derived, native ras-related polypeptide Yp20 . Only 2% (1 of 58) of normal control sera showed strong competition, as compared to 15% (5 of 34) of patients with early stage or no evidence of disease, and 44% (26 of 59) of patients with advanced disease . These differences were statistically significant (x2, P less than 0.05-0.001) . Eleven sera samples of cancer patients found to be strong competitors in the ELISA competition kit were tested for the presence of anti-ras antibodies by ELISA . None showed higher ELISA values as compared with pooled normal human serum and control sera . It is thus suggested that our procedure detected circulating ras-related onco-proteins in sera of cancer patients mainly with advanced disease.

J Assoc Off Anal Chem, 1989 Jan-Feb, 72(1), 30 - 3
Determination of aspartame in beverages using an alcohol oxidase enzyme electrode; Smith VJ et al.; A new method for the determination of the artificial sweetener aspartame is described . alpha-Chymotrypsin is used to cleave the methyl ester group of aspartame, producing methanol hydrolytically . The methanol is detected using an electrode which is constructed by physically trapping yeast alcohol oxidase enzyme at the tip of a dissolved oxygen electrode . The decrease in oxygen concentration, which occurs as methanol is enzymatically oxidized to formaldehyde, is measured amperometrically . Aspartame levels in diet soft drinks as determined by the proposed method and by liquid chromatography are in excellent agreement . The relative standard deviation of the measurements is 0.83% . The methanol present in diet cola as a result of aspartame degradation can also be measured by using the electrode without alpha-chymotrypsin.

Agents Actions, 1989 Jan, 26(1-2), 258 - 60
Evaluation of the functional integrity of endothelium in perfused lungs; Bult H et al.; Prostacyclin (PGI2) formation and the saturable uptake of 5-hydroxytryptamine (5HT) were studied as indices of endothelial integrity in isolated lungs . 5HT uptake was characterized by its kinetic parameters Km and Vmax . These were calculated from multiple indicator dilution data on the basis of an organ model of 5HT uptake . Perfused dog lung lobes were exposed to plasma activated with yeast (YAP) or zymosan (ZAP) . YAP induced a transient elevation of Km . This increase probably reflects endothelial injury . Vmax remained unchanged, suggesting that the perfused endothelial surface remained stable . PGI2 biosynthesis was negligible in the control period, but started immediately after exposure to ZAP or YAP . It was proportional to the transient elevation of Km and to the pulmonary oedema . The data suggest that PGI2 might be a marker of severe endothelial distress.

Rev Environ Contam Toxicol, 1989, 108, 105 - 32
Cobalt in the environment and its toxicological implications; Domingo JL; Cobalt is an essential trace element which is widely distributed in nature . Most of cobalt consumed is used in the manufacture of alloys, and although not released extensively in the environment, it may represent a hazard to human health . In addition, excess dietary cobalt produces toxic effects in animals . Polycythemia and hyperglycemia with transitory damage to pancreatic alpha-cells have been widely reported after cobalt administration . Cobalt salts induce respiratory deficiency in yeast . CoCl2 increased sister chromatid exchange (SCE) in P388D1 cells and in lymphocytes from two donors . So far it has not been possible to induce cancer in experimental animals using cobalt by any other route than by injection . Ingestion of cobalt may lead to reproductive changes in the male rat such as loss of testicular volume and darkening of testicle color . On the other hand, oral administration of cobalt did not produce teratogenicity or significant fetotoxicity in the rat at daily doses as high as 100 mg CoCl2/kg . However, cobalt affected the period of late gestation as well as the postnatal development of the pups . Occupational toxicology of cobalt, hygienic and epidemiologic aspects, and treatment of cobalt poisoning are also topics of special interest . Cobalt is a metal with marked allergenic potential . Asthma, interstitial lung disease and combined asthma and alveolitis have been described as occupational health hazards . EDTA, DTPA, and N-acetyl-L-cysteine have been suggested as possible antidotes in cobalt intoxication.

Carlsberg Res Commun, 1989, 54(5), 193 - 202
Primary structure of carboxypeptidase III from malted barley; Sorensen SB et al.; The primary structure of malt carboxypeptidase III has been determined . The enzyme is a single N-terminally blocked polypeptide chain containing 411 amino acid residues . The sequence of these amino acid residues was deduced from analysis of fragments of the polypeptide chain obtained by chemical cleavages with either cyanogen bromide or hydroxylamine and by enzymatic cleavages with either trypsin, S . aureus V8 protease or proteinase A from yeast . A glycosylated asparagine was found in position 71 . The determined sequence was 97% homologous with the amino acid sequence derived from the nucleotide sequence of a gene coding for a wheat protein postulated to be a carboxypeptidase . The malt carboxypeptidase III sequence showed 34% homology with the amino acid sequence of the single-chain carboxypeptidase Y, and about 25% homology with the combined A- and B-chains of malt carboxypeptidase I and II as well as wheat carboxypeptidase II.

Cell Motil Cytoskeleton, 1989, 14(4), 544 - 51
Kinetics of granulocyte phagocytosis: rate limited by cytoplasmic viscosity and constrained by cell size; Evans E; Micromanipulation of yeast particles and blood granulocytes has been used to study the kinetics of single phagocytosis events . The ingestion process was quantitated by observation of sequential adhesion and encapsulation times . Both adherence and encapsulation times were found to increase greatly as the temperature was reduced below 37 degrees C; calcium in solution facilitated adhesion of the particle to the phagocyte but not encapsulation; both adhesion and encapsulation processes required a minimum level of plasma components (presumably complement) . The general nature of these observations were confirmatory of previous studies, but this study is unique in that the specific time course of single particle ingestion was quantitated . It was immediately apparent that the phagocytosis process was 100% efficient above the threshold concentrations required for plasma and temperature, but variations in times from cell to cell indicated heterogeneity in the population . The total time for ingestion varied from as low as 2 sec/particle at 37 degrees C to above several min/particle below 15 degrees C . Encapsulation times for particles were normalized by estimates of particle surface areas to establish a specific time/unit area of particle surface: from 0.5 sec/10(-8) cm2 at 37 degrees C to greater than 8 sec/10(-8) cm2 at 15 degrees C . The temperature dependence of the encapsulation time correlated well with the temperature dependence of the "apparent" viscosity for granulocytes measured by micropipet aspiration . As such, the kinetic properties observed in these phagocytosis tests are consistent with a model that both assembly of the contractile system and the displacement of the surface by active contraction in phagocytosis are limited by viscous dissipation in the cell.(ABSTRACT TRUNCATED AT 250 WORDS)

Adv Enzyme Regul, 1989, 28, 219 - 36
Patterns of divergence in homologous proteins as indicators of tertiary and quaternary structure; Benner SA; A new approach for extracting conformational information from an alignment of homologous proteins is presented . This approach extracts information from the pattern of sequence divergence in proteins, and considers evolutionary issues, such as functional adaptation and neutral drift, in assigning roles in tertiary structure to residues at specific positions in the alignment . A reliable algorithm is developed for identifying surface residues in a protein . An algorithm is also developed for identifying active site residues; this algorithm can be applied in cases where functional divergence occurs in one subgroup of homologous proteins but not in others . Finally, these algorithms are used to make predictions regarding the quaternary structure of alcohol dehydrogenase from yeast.

Eur Biophys J, 1989, 17(4), 233 - 5
Effect of aminoacylation on tRNA conformation; Antosiewicz J et al.; Translational diffusion coefficients have been simulated for various conformations of tRNA(Phe) (yeast) by bead models, in order to analyze data obtained by dynamic light scattering on the free and the aminoacylated form . The 18% increase of the translational diffusion coefficient upon deacylation, reported by Potts et al . (1981), could not be represented by any change of the L-hinge angle, but could only be simulated by a conformation change to an extended form with extensive dissociation of base pairs . Since extensive unpairing is not consistent with evidence accumulated in the literature, the change of the diffusion coefficient must be mainly due to processes other than intramolecular conformational changes.

J Med Vet Mycol, 1989, 27(5), 335 - 41
Mel- mutants of Wangiella dermatitidis in mice: evaluation of multiple mouse and fungal strains; Dixon DM et al.; Melanin-deficient mutants of Wangiella dermatitidis were studied by comparing a spontaneous mutant (Mel 3) with the parental wild type (wt) in four strains of mice and by comparing various UV-induced mutants in a single strain of mouse . Mice were inoculated intravenously with 3 x 10(6), 1 x 10(7), 3 x 10(7) or 1 x 10(8) yeast-like cells and mortality assessed at the end of 20 days . Neuropathology was evaluated in methenamine-silver stained brain sections of the different strains of mice injected with either wt or Mel 3 (1 x 10(7) cells per mouse) . In general, both fungus strain related and mouse strain related differences in mortality were observed . The DBA/2J mouse was the most susceptible to fatal infection with all fungus strains . In the other strains of mice, however, the melanin deficient mutants were significantly less virulent than the wt at a concentration of less than or equal to 3 x 10(7) cells per mouse . No obvious trend was seen in the numbers of brain lesions in different strains of mice with respect to wt vs . Mel 3 . However, invasive hyphal forms did seem to be associated with virulence.

Acta Derm Venereol, 1989, 69(4), 359 - 62
The effect on atopic dermatitis of supplementation with selenium and vitamin E; Fairris GM et al.; Reduced concentrations of selenium in whole blood, plasma and white cells and reduced activity of selenium-dependent glutathione peroxidase in red cells have been found in atopic dermatitis . To determine the effect of selenium supplementation on this disease, the normal daily diet of 60 adults with atopic dermatitis was supplemented with selenium-enriched yeast for 12 weeks in a randomised double-blind study . Group 1 took 600 micrograms of selenium alone, Group 2 600 micrograms of selenium plus 600 IU of vitamin E and Group 3 a placebo . After 12 weeks, there was a significant increase in the concentration of selenium in whole blood and the activity of selenium dependent glutathione peroxidase in platelets in Groups 1 and 2 and the concentration of vitamin E in plasma in Group 2 . There was no significant difference between the three Groups in the severity of the eczema or the concentration of selenium either before or after the 12 weeks of supplementation . The results suggest that although selenium-enriched yeast supplement was absorbed and bioavailable it does not enter the skin or produces a worthwhile improvement in atopic dermatitis.

Australas J Dermatol, 1989, 30(1), 37 - 40
Phaeohyphomycosis due to Phialophora richardsiae; Tam M et al.; Phaeohyphomycosis, an infection characterised by dematiaceous yeast-like cells, hyphae and pseudohyphae in tissue, is an uncommon condition, often affecting immunosuppressed patients . A sixty four year old boat-builder, receiving treatment with prednisone and azathioprine developed multiple cutaneous nodules on the extremities . Histology showed a mixed dermal inflammatory infiltrate with scattered spores and hyphae . Culture revealed two organisms, Phialophora richardsiae and Exophiala jeanselmei . Fluorocytosine was initially given but the organism was found to be resistant . Since side effects have been associated with long term ketoconazole therapy, a less toxic and more potent triazole compound, itraconazole, was used . After three months, the lesions had completely resolved without adverse clinical or biochemical changes.

J Hyg Epidemiol Microbiol Immunol, 1989, 33(4), 397 - 408
Viral contamination of some drinking waters in Romania: a twenty years survey; Nestor I et al.; A synthesis is made on the 20 years virological survey of the drinking water from some towns of Romania . The sampling of the water was made by the gauze-pad method . The virus concentration method by adsorption-elution with the yeast cells was applied concomitantly, at first with the PE60 method, then, during the last years, either with aluminum hydroxide or with the polymer PV methods, and the concentrates were inoculated both into suckling mice and into cell cultures . Various types of coxsackievirus A and B, poliovirus and adenovirus were detected . The proportions of positive samples varied between 0 and 25% annually, in diverse towns, the mean proportion being 2.1% . These proportions are relative low, although two concentrating and two detecting methods were applied concomitantly.

Prog Clin Biol Res, 1989, 297, 287 - 96
Phagocytosis of particulate activators of the human alternative complement pathway through monocyte beta-glucan receptors; Czop JK et al.; Human monocytes phagocytose particulate activators of the alternative complement pathway through beta-glucan receptors in the absence of opsonins . Recognition of soluble beta-glucans by monocytes selectively inhibits ingestion of particulate activators and has no effect on responses mediated by monocyte receptors for Fc-IgG, complement, or fibronectin . The smallest ligand unit recognized by monocyte beta-glucan receptors is an acid-resistant heptaglucoside present in yeast cell walls . Mouse monoclonal anti-beta-glucan antibodies have been prepared, one of which completely neutralizes the inhibitory capacity of the HPLC-purified heptaglucoside . This antibody has been used as immunogen for the preparation of an anti-Id . The pretreatment of monocytes with low concentrations of anti-Id inhibits monocyte ingestion of zymosan particles but not EsIgG, suggesting that this antibody has specificity for monocyte beta-glucan receptors and is a powerful probe for further receptor studies . Other receptors with specificities for carbohydrates are also present on mononuclear phagocytes . Receptors for mannose/fucose and those for galactose have been isolated and cloned . The development of probes, such as structural analogs of the active heptaglucoside and the anti-Id, will bring the beta-glucan receptors to a similar stage of definition . A major factor that adds a considerable degree of difficulty to studies of the beta-glucan receptors and is not shared by the other receptors for carbohydrates is the requirement for structural conformations provided by the alignment of several glucose units rather than the recognition of a hexose residue in, for example, a glycoconjugate . The designation of these receptors as beta-glucan receptors has inadvertently taken us into a new area of nonimmune defense . Animal studies indicate that beta-glucans with 1,3- and/or 1,6-linkages are active pharmacologic agents that rapidly confer protection to a normal host against a variety of biologic insults . The beta-glucan receptors provide a mechanism by which a heightened state of host responsiveness is initiated.

Bull Soc Pathol Exot Filiales, 1989, 82(4), 451 - 7
{Cryptosporidium and candida in pediatric diarrhea in Abidjan}; Koffi-Akoua G et al.; In a study involving 104 children hospitalized with diarrhoea, 9% were infected with oocyst Cryptosporidium spp . add 56% with such yeast-fungus as Candida (C . Candida 38%) . The manifestations noted in cryptosporidiosis infected children are acute diarrhea, vomiting and hyperthermia . One subject out of five who were tested for antibody to HIV appeared to be antibody positive . The patients immunity from the disease was not checked . A mycological test must be systematically carried out in case of children diarrheal outbreak.

Cell Motil Cytoskeleton, 1989, 14(4), 485 - 90
Purification of anterogin, a protein factor necessary for the dispersion of carotenoid droplets in permeabilized xanthophores of goldfish; Zeng ZC et al.; We reported previously that the dispersion of carotenoid droplets in permeabilized xanthophores requires cAMP, ATP, and a cytosolic factor present in several secretory tissues as well as in xanthophores . We have now purified this factor from beef liver to apparent/near homogeneity . It appears to be a heterodimer with Mr approximately 125,000 . The purified factor has little or no ATPase activity, with or without the presence of actin . Nor does it stimulate the ATPase activity of carotenoid droplets . Its exact function in carotenoid droplet dispersion is thus unclear . Since dispersion of carotenoid droplets is an anterograde translocation, we propose the name anterogin for this protein . We also report that yeast cytosol has anterogin activity.

Allergol Immunopathol (Madr), 1989 Jan-Feb, 17(1), 39 - 43
Monocyte functions in patients with mixed connective tissue disease; Bodolay E et al.; The authors investigated the number and functions of peripheral blood monocytes in patients with mixed connective tissue disease . Moderate monocytopenia was detected in the active stage of the disease . There was a close correlation between the sensitized sheep red blood cell binding capacity of monocytes and the elevated levels of circulating immune complexes . The yeast phagocytosis of monocytes and the chemotactic activity were normal in patients with mixed connective tissue disease . C3b receptor mediated phagocytosis of monocytes decreased in the active and inactive stages of the disease . This study suggests that the decreased function of C3b receptors of monocytes is related to the pathogenesis of this disease.

Bull World Health Organ, 1989, 67(1), 65 - 70
The hepatitis B immunization programme in Singapore; Goh KT et al.; A voluntary immunization programme to prevent perinatal transmission of hepatitis B virus (HBV) infection in Singapore was implemented on 1 October 1985 as an integral component of the national childhood immunization programme . Up to April 1988, a total of 68,845 mothers who attended government maternal and child health clinics were screened for the disease . Of these, 2432 (3.5%) were positive for hepatitis B surface antigen (HBsAg) and 904 (1.3%) for hepatitis B e antigen (HBeAg) . Virtually all the babies born to carrier mothers completed the full immunization schedule; and in addition, those of HBeAg-positive mothers were given a dose of hepatitis B immunoglobulin at birth . The hepatitis B immunization programme was extended on 1 September 1987 to cover all newborns . About 90% of the 15,943 babies delivered in government institutions from September 1987 to April 1988 were immunized at birth, with the subsequent doses being administered at maternal and child health clinics at 4-6 weeks and 5 months later . More than 85% of the children given the full course of plasma-derived and yeast-derived hepatitis B vaccine from birth continued to have protective antibody to HBV two years after immunization . The programme is being closely monitored to assess the duration of immunity and the need for booster doses, while seronegative adults are also being encouraged to be vaccinated.

Int J Immunopharmacol, 1989, 11(8), 855 - 62
Enhanced killing of Blastomyces dermatitidis by gamma interferon-activated murine peripheral blood polymorphonuclear neutrophils; Morrison CJ et al.; Normal peripheral blood polymorphonuclear neutrophils (PB-PMNs), challenged in vitro with yeast form Blastomyces dermatitidis, reduced inoculum colony-forming units of a virulent strain by 37.5 +/- 9.5% . Pre-incubation of PB-PMNs with 10-100,000 U/ml of purified recombinant murine gamma-interferon (IFN) for 1 h prior to challenge with fungi resulted in significant enhancement of PB-PMN fungicidal activity . No direct fungicidal activity by IFN alone was observed . Pretreatment of selected concentrations of IFN shown to have PMN-enhancing activity (100 or 1000 U/ml) with rabbit hyperimmune anti-IFN antiserum for 1 h before addition to PB-PMNs abrogated the enhancement of fungicidal activity . Isolated peripheral blood mononuclear cells failed to kill B . dermatitidis, even when mononuclear cells were present at a concentration ten times greater than that normally used in killing assays, and failed to be activated by IFN . Treatment of unstimulated or IFN-activated PB-PMNs with complement and hybridoma-derived monoclonal antibody specific for PMNs eliminated PB-PMN fungicidal activity . Exogenously added lipopolysaccharide (0.0005-50,000 ng/ml) did not activate PB-PMNs, whether added alone or in conjunction with IFN . The PB-PMN activating capacity of IFN could be destroyed by heat treatment (100 degrees C, 15 min) or by acid treatment with HCl (pH 2) . These results demonstrate that recombinant gamma-interferon can stimulate PB-PMNs to kill B . dermatitidis, that the PB-PMN activating moiety is IFN and that PB-PMNs are responsible for fungal killing in this assay system.(ABSTRACT TRUNCATED AT 250 WORDS)

Folia Microbiol (Praha), 1989, 34(3), 238 - 42
Scanning electron microscope study of Torulopsis ethanolitolerans prepared by glutaraldehyde--thiosemicarbohydrazide procedure; Hulinska D et al.; Torulopsis ethanolitolerans subject to both the sparing and coarse heat treatment were studied in the scanning electron microscope . The reduction of adhesivity, increased permeability and higher rigidity of the yeast wall was achieved by an original glutaraldehyde-paraformaldehyde fixation, low osmolarity in vacuo and subsequent thiosemicarbohydrazide incubation, followed by addition of metal salt . The impregnation of the metal throughout the specimen due to the reaction of the thiosemicarbohydrazide with glutaraldehyde allowed viewing of small or intricate surface details of the yeast . Structural differences of the yeast processed by sparing and coarse heat treatment were shown to be better from the thiosemicarbohydrazide incubated samples compared to those that were prepared with osmium tetroxide.

J Protozool, 1989 Jan-Feb, 36(1), 29 - 34
Codon usage in Tetrahymena and other ciliates; Martindale DW; Codon usage in ciliates was examined by analyzing the coding regions of 22 ciliate genes corresponding to a total of 26,142 nucleotides (8,714 codons) . It was found that Tetrahymena, Paramecium and the hypotrichs (Oxytricha and Stylonychia) differed in which synonymous codons were used most frequently by their genes . In fact, the codon choices in highly expressed Tetrahymena genes were more similar to those of yeast genes than those of Paramecium genes . The ciliates do not appear to have unusually strong biases in codon usage frequency when compared to other protists such as yeast . The analysis of the Tetrahymena genes indicated that genes which are highly expressed during normal cell growth have a stronger bias towards using the "preferred" codons than those expressed at lower levels during growth or for brief periods during processes such as conjugation . This conforms to what is found in other protists.

Chem Biol Interact, 1989, 69(2-3), 193 - 201
Effects of neonatal monosodium-L-glutamate treatment on rat alveolar macrophages; Liu WK et al.; Treatment of neonatal rats with repeated doses of monosodium-L-glutamate resulted in changes of morphology and function of alveolar macrophages recovered from adult female rats . Numerous cellular lipid vacuoles and lamellar structures were observed in these alveolar macrophages under transmission electron microscopy . There was approximately 50% increase of cellular total lipid content measured by Oil-Red-O staining and colorimetric method . The yeast phagocytosis and intracellular killing ability, as well as the inhibitory effect on the growth of tumor cells were reduced in alveolar macrophages from neonatal MSG-treated rats.

Life Sci, 1989, 44(26), 2067 - 74
RNA-induced transformation of the estrogen receptor detected by a monoclonal antibody which recognizes the activated receptor; Giambiagi N et al.; The effect of RNA and polyribonucleotides on the estrogen receptor from fetal guinea pig uterus was studied through the analysis of the sedimentation properties of this receptor and its interaction with the monoclonal antibody D547 . Different exogenous RNAs (calf thymus RNA, yeast RNA and rabbit liver transfer RNA) were able to induce a transformation of the 9S native receptor to 4.5-7S sedimenting forms in low salt sucrose density gradients, as activating factors such as temperature and time do . This transformation was prevented by 20mM sodium molybdate . Moreover, the RNA treated receptor was partially recognized by the monoclonal antibody D547 . This antibody, as was demonstrated previously, selectively reacts with the activated form of this receptor . When different homo-polyribonucleotides were tested, the effect depended on their composition . In contrast, DNA did not affect either the sedimentation properties of the receptor or its reaction with the antibody . These observations suggest that RNA induces a dissociation of the 9S receptor and that at least one of the resulting forms is the activated receptor . However, RNA and polyribonucleotides inhibited the receptor binding to DNA-cellulose apparently by competing with DNA . The data suggest a role of RNA in estrogen receptor activation.

Blood, 1989 Jan, 73(1), 90 - 9
The in vivo metabolism of recombinant human erythropoietin in the rat; Spivak JL et al.; We compared the in vivo plasma clearance and organ accumulation in anesthetized rats of 125I-labeled, recombinant human erythropoietin and 125I-labeled, desialylated recombinant erythropoietin . The immediate volume of distribution of 125I-labeled, recombinant erythropoietin approximated that of the plasma volume . Its plasma clearance was multiexponential, with an initial rapid distribution phase (t1/2 = 53 minutes) and a slower elimination phase (t1/2 = 180 minutes) . Organ accumulation of labeled recombinant erythropoietin, as compared with 125I-labeled human albumin, was negligible until 30 minutes after injection when small amounts appeared in the kidneys and bone marrow . Only 24% of the 125I-labeled, desialylated recombinant erythropoietin was recovered immediately after injection, and 96% of the hormone was cleared from the plasma with a t1/2 of 2.0 minutes . The bulk of the desialylated hormone accumulated in the liver where it was rapidly catabolized and its breakdown products released back into the plasma . Significantly, in contrast to unmodified erythropoietin, there was also early accumulation of desialylated hormone in the kidneys, marrow, and spleen . Desialylated orosomucoid but not orosomucoid, yeast mannan, or dextran sulfate 500 inhibited the rapid plasma clearance and hepatic accumulation of desialylated erythropoietin . Oxidation of the desialylated hormone restored its plasma recovery and clearance to normal but rendered it biologically inactive, and accumulation in organs other than the kidney was negligible.

J Biol Chem, 1988 Dec 25, 263(36), 19833 - 42
Degradation of structurally characterized proteins injected into HeLa cells . Basic measurements; Rogers SW et al.; Thirty-five proteins of known x-ray structure were labeled by chloramine-T radioiodination or by reaction with 125I-Bolton-Hunter reagent and introduced into HeLa cells using red cell-mediated microinjection . Degradation rates of the injected proteins were then determined over the next 50 h by measuring the release of soluble isotope to the culture medium . Control experiments demonstrated that the measured rates were not compromised by proteolysis within RBCs, the presence of unfused RBCs, or degradation of protein released from RBCs to the medium . Degradation of some injected proteins was faster during the first 12 h after fusion than at later times, apparently a response of HeLa cells to trypsinization . However, all proteins exhibited first-order degradation rates between 24 and 48 h post injection . Except for seven proteins, stabilities measured during this interval were unaffected by the labeling procedure . Reductive methylation was used to choose among the seven discordant values, and half-lives for the 35 proteins ranged from 16 h for lysozyme to 214 h for yeast alcohol dehydrogenase . Since half-lives for six of the injected proteins closely match values obtained by in vivo measurements, we consider our estimates of the metabolic stabilities of the injected proteins to be generally accurate . Therefore, the half-lives obtained by microinjection should prove useful in the search for relationships between protein structure and intracellular stability.

Cell, 1988 Dec 23, 55(6), 989 - 1003
Isolation and properties of cDNA clones encoding SRF, a transcription factor that binds to the c-fos serum response element; Norman C et al.; The serum response element (SRE) is a sequence required for transient transcriptional activation of genes in response to growth factors . We have isolated cDNA clones encoding serum response factor (SRF), a ubiquitous nuclear protein that binds to the SRE . The SRF gene is highly conserved through evolution, and in cultured cells its transcription is itself transiently increased following serum stimulation . A cDNA clone of SRF expressed in vitro generates protein that forms complexes indistinguishable from those formed with HeLa cell SRF, as judged by DNA binding specificity and the ability to promote SRE-dependent in vitro transcription . SRF binds DNA as a dimer, and the DNA binding/dimerization domain of the protein exhibits striking homology to two yeast regulatory proteins.

J Biol Chem, 1988 Dec 15, 263(35), 19092 - 7
Phosphorylation and inactivation of the pyruvate dehydrogenase from the anaerobic parasitic nematode, Ascaris suum . Stoichiometry and amino acid sequence around the phosphorylation sites; Thissen J et al.; Tryptic digestion of the fully phosphorylated Ascaris suum pyruvate dehydrogenase complex yielded a single tetradecapeptide containing 2 phosphorylated serine residues . Its amino acid sequence was Tyr-Ser-Gly-His-Ser(P)-Met-Ser-Asp-Pro-Gly-Thr-Ser(P)-Tyr-Arg and was very similar to one of the tryptic phosphopeptides isolated from mammalian and yeast pyruvate dehydrogenases . At partial phosphorylation, three peptides were isolated which corresponded to the monophosphorylated (sites 1 and 2) and diphosphorylated tetradecapeptides . In contrast to results reported from mammalian complexes, phosphorylation of the ascarid complex paralleled inactivation, and no additional phosphorylation occurred after inactivation was complete . Complete inactivation of the complex was associated with the incorporation of 1.7-1.9 mol of phosphoryl groups/mol of alpha-pyruvate dehydrogenase subunit, and the strict preference of the pyruvate dehydrogenase kinase for site 1 was not observed . Whereas site 1 was initially phosphorylated more rapidly than site 2, at 50% inactivation, 41% of the incorporated phosphoryl groups were incorporated into site 2 . In addition, substantial amounts of peptide monophosphorylated at site 2 also accumulated, suggesting that prior phosphorylation at site 1 was not necessary for phosphorylation at site 2 . Phosphorylation also caused a marked decrease in the mobility of the alpha-pyruvate dehydrogenase subunit on sodium dodecyl sulfate-polyacrylamide gels and the apparent separation of mono- and diphosphorylated forms of the enzyme . The significance of these observations in the regulation of the unique anaerobic mitochondrial metabolism of A . suum is discussed.

Science, 1988 Dec 9, 242(4884), 1430 - 3
Cyclic AMP-responsive DNA-binding protein: structure based on a cloned placental cDNA; Hoeffler JP et al.; Cyclic AMP (cAMP) is an intracellular second messenger that activates transcription of many cellular genes . A palindromic consensus DNA sequence, TGACGTCA, functions as a cAMP-responsive transcriptional enhancer (CRE) . The CRE binds a cellular protein of 38 kD in placental JEG-3 cells . A placental lambda gt11 library was screened for expression of specific CRE-binding proteins with the CRE sequence as a radioactive probe . A cDNA encoding a protein of 326 amino acids with the binding properties of a specific CRE-binding protein (CREB) was isolated . The protein contains a COOH-terminal basic region adjacent to a sequence similar to the "leucine zipper" sequence believed to be involved in DNA binding and in protein-protein contacts in several other DNA-associated transcriptional proteins including the products of the c-myc, c-fos, and c-jun oncogenes and GCN4 . The CREB protein also contains an NH2-terminal acidic region proposed to be a potential transcriptional activation domain . The putative DNA-binding domain of CREB is structurally similar to the corresponding domains in the phorbol ester-responsive c-jun protein and the yeast transcription factor GCN4.

Cell, 1988 Dec 2, 55(5), 899 - 906
Multiple and cooperative trans-activation domains of the human glucocorticoid receptor; Hollenberg SM et al.; A 30 amino acid peptide (referred to as tau 2) that functions as an activation domain has been localized in the carboxyl terminus of the human glucocorticoid receptor . This sequence, when fused to yeast GAL4 as part of the ligand binding domain, generates a hormone-inducible activator . Tau 2 functions in a position-independent fashion and leads to a progressive gain of function when multimerized . A similar and independent activity has also been identified in the amino terminus of the receptor . These two sequences, although structurally unrelated, are both acidic in character and thus may have certain properties in common with yeast activator sequences.

Cell, 1988 Dec 2, 55(5), 785 - 95
A deduced gene product from the Drosophila neurogenic locus, enhancer of split, shows homology to mammalian G-protein beta subunit; Hartley DA et al.; The correct segregation of neural from epidermal lineages in Drosophila embryogenesis depends on the activity of the six zygotic "neurogenic" genes . One of the neurogenic genes, Enhancer of split, is particularly noteworthy in its genetic interactions with Notch and Delta, which both appear to code for transmembrane proteins with homology to the epidermal growth factor . Transformation experiments have demonstrated the cloning of sequences necessary for Enhancer of split gene function . We report here that the gene product derived from DNA sequencing shows homology to the beta subunit of mammalian G proteins and CDC4, a yeast cell cycle gene . We demonstrate that expression of the transcripts relates to the developing central nervous system . These data suggest a mechanism of interaction between the gene products of Notch and Enhancer of split.

Mycopathologia, 1988 Dec, 104(3), 181 - 8
In situ localization of antigens of Histoplasma capsulatum using colloidal gold immune electron microscopy; Graybill JR et al.; Histoplasma capsulatum contains multiple antigens, among them the H antigen and M antigen, which are useful in serologic testing for histoplasmosis . We prepared 7 mouse monoclonal antibodies (5 IgG, 2 IgM) to histoplasmin, and compared these with polyclonal histoplasmin antibodies raised in rabbits and mice . Both monoclonal and polyclonal antibodies were high titered by ELISA . Colloidal gold immune electron microscopy (CGIEM) showed that polyclonal antibodies to histoplasmin or H antigen bound at multiple sites in the cell wall, cytoplasm, and nucleus of Histoplasma yeast cells . In contrast, antibodies to M antigen selectively label the cell membrane and antibodies to alkali soluble cell wall antigen label only the cell wall . Polyclonal antibodies cross reacted extensively with other fungi, both by ELISA and CGIEM . Monoclonal antibodies stained only cytoplasmic epitopes, but also cross reacted with other fungi by electron microscopy . Only periodate treated H antigen elicited polyclonal antibodies which were more specific than those of untreated H antigen or histoplasmin.

Biochem J, 1988 Dec 1, 256(2), 571 - 7
N.m.r., e.p.r . and magnetic-c.d . studies of cytochrome f . Identity of the haem axial ligands; Rigby SE et al.; N.m.r.-, magnetic-c.d.- and e.p.r.-spectroscopic studies of oxidized and reduced cytochrome f from charlock, rape and woad are reported . Comparison of the spectra with corresponding spectra of other haem proteins, including horse and yeast cytochromes c, bovine cytochrome b5 and n-butylamine adduct of soya-bean leghaemoglobin support the hypothesis {Siedow, Vickery & Palmer (1980) Arch . Biochem . Biophys . 203, 101-107} that lysine is the sixth ligand of native cytochrome f . Detailed analysis of the e.p.r . spectrum of ferricytochrome f indicates that its principle g-values are 3.51, 1.70 and less than 1.3, and not 3.48, 2.07 and 1.6 as previously suggested {Siedow, Vickery & Palmer (1980) Arch . Biochem . Biophys . 203, 101-107} . The observation of a one-proton intensity resonance at -3.27 p.p.m . in the 1H-n.m.r . spectrum of ferrocytochrome f, coupled with the absence of a methionine methyl resonance from the spectral region to low frequency of -2 p.p.m., is suggested to be a general indicator of lysine co-ordination.

EMBO J, 1988 Dec 1, 7(12), 3823 - 8
Wheat germ splicing endonuclease is highly specific for plant pre-tRNAs; Stange N et al.; Intron-containing pre-tRNAs from organisms as different as yeast, Nicotiana, Xenopus and man are efficiently spliced and processed in a HeLa cell extract . They are also correctly processed in a wheat germ extract; however, the intron is removed only from the tobacco pre-tRNA . To determine whether plant pre-tRNA introns have any specific structural and/or sequence feature we have cloned two intron-containing tRNATyr genes from the plant Arabidopsis . Comparison of these genes, of the Nicotiana tRNATyr gene and of a Glycine max tRNAMet gene reveals that plant introns from three different species have no sequence homology and are only 11 to 13 nucleotides long . Thus, short length may be one important feature of plant introns . Furthermore, the 5' and 3' splice sites are separated by 4 bp in the extended anticodon stems of these pre-tRNA structures . In contrast, yeast and vertebrate introns are rather variable in length and the splice sites are separated by 5 or 6 bp . These differences in distance and relative helical orientation of the splice sites in plant pre-tRNAs versus pre-tRNAs from other organisms are obviously tolerated by the vertebrate splicing endonuclease, but not at all by the plant enzyme.

Proc Natl Acad Sci U S A, 1988 Dec, 85(23), 8815 - 9
Metal-specific synthesis of two metallothioneins and gamma-glutamyl peptides in Candida glabrata; Mehra RK et al.; Cellular resistance to heavy metal cytotoxicity in most species is mediated by the binding of metal ions either to a cysteine-rich polypeptide in the metallothionein family or to short cysteine-containing gamma-glutamyl peptides . One of these metal binding systems has been found in most organisms studied . However, the yeast Candida (Torulopsis) glabrata expresses both metallothionein and the gamma-glutamyl peptides for metal detoxification, and each system is regulated in a metal-specific manner . Exposure of C . glabrata to copper salts stimulates formation of two metallothionein-like polypeptides with a cysteine content of 30 mol% and the repeated sequence Cys-Xaa-Cys . The cells synthesize gamma-glutamyl peptides upon exposure to cadmium salts . Penta- and tetrapeptides that form a cadmium-thiolate cluster in a peptide oligomer containing labile sulfur are synthesized.

Arch Pathol Lab Med, 1988 Dec, 112(12), 1262 - 4
Disseminated penicilliosis in a patient with acquired immunodeficiency syndrome; Piehl MR et al.; We describe a case of disseminated penicilliosis in a patient with the acquired immunodeficiency syndrome . Penicillium marneffei was cultured from the blood, bone marrow, sputum, stool, and skin; the yeast forms were demonstrated in skin and bone marrow biopsy specimens . To our knowledge, this is the first reported case of disseminated penicilliosis described in a patient with the acquired immunodeficiency syndrome . The differential diagnosis with Histoplasma capsulatum is reviewed.

Am J Hematol, 1988 Dec, 29(4), 211 - 4
Immunogenicity of a recombinant hepatitis B vaccine in hemophiliacs; Mannucci PM et al.; Yeast-recombinant vaccines against hepatitis B virus (HBV) are now available, but there is no information about whether or not they are immunogenic in patients with hemophilia and other congenital bleeding disorders . Twenty micrograms of a recombinant vaccine expressing the adw serotype of the hepatitis B surface antigen (HBsAg) were given to 41 patients negative for HBV markers and again after 1 and 6 months . Ten percent of the vaccinees had anti-hepatitis B surface antibody (anti-HBs) responses, with titers of 10 mIU/ml or more, 1 month after the first dose of vaccine . The percentage of anti-HBs-positive patients increased to 54% after the second dose and to 98% after the third dose, with only one non-responder . Hence, the recombinant vaccine was immunogenic, with percentages of seroconversion and anti-HBs titers similar to those achieved with plasma-derived vaccines.

Infect Immun, 1988 Dec, 56(12), 3121 - 5
Virulence conversion of Legionella pneumophila: a one-way phenomenon; Catrenich CE et al.; Previous investigations have shown that Legionella pneumophila converts from virulence to avirulence after passage on supplemented Mueller-Hinton (SMH) agar and may convert back to virulence after passage in guinea pigs . However, there is no additional information concerning the apparent interconversion of virulent and avirulent derivatives of L . pneumophila cultures . We investigated the stability of a parental virulent culture and its avirulent derivatives and the growth and viability of these cultures on charcoal-yeast extract (CYE) and SMH agars . Avirulent derivatives of a highly virulent L . pneumophila culture were obtained by passage of the virulent parent culture on SMH agar . The only time a virulent L . pneumophila culture was recoverable from an avirulent culture was when the avirulent culture was derived from a saline suspension of a virulent culture which had been passaged only five times on SMH agar . When an avirulent culture was derived from a virulent culture passaged 25 times on SMH agar or from an isolated colony which grew on a SMH agar plate, we were unable to recover a virulent culture after successive passage through guinea pigs . These results suggest that the conversion process which occurs between virulent and avirulent forms of L . pneumophila is a one-way phenomenon from virulence to avirulence and that stable avirulent derivatives can be isolated . Furthermore, our findings suggest that SMH agar acts as a selective medium for the growth of avirulent L . pneumophila, and growth on SMH agar may be a phenotypic marker for avirulence . Virulent cells, although unable to grow on SMH agar, may remain viable for several passages on SMH agar and propagate when inoculated into guinea pigs.

Cancer Res, 1988 Dec 1, 48(23), 6715 - 20
Investigation of the effects of heat shock and agents which induce a heat shock response on the induction of differentiation of HL-60 cells; Richards FM et al.; A heat shock of 42.5-43.5 degrees C for 1 h applied to HL-60 promyelocytic leukemia cells induced the appearance of between 13 and 34% (n = 6) of cells which showed characteristics of mature metamyelocytes/granulocytes . This is the first time a physical agent has been shown to induce the differentiation of this leukemic cell line . The treatment of HL-60 cells with a variety of agents which have been documented to stress cells and induce thermotolerance or a heat shock-like response also induced granulocyte-like differentiation: continuous treatment for 4 days with ethanol (213 mM), sodium arsenite (6 microM), cadmium sulfate (60 microM), lidocaine (3 mM), and procaine (5 mM) induced 73, 54, 14, 54, and 55% of cells, respectively, to reduce the dye nitro blue tetrazolium . They were also capable of the phagocytosis of yeast particles . Examination of differentiated cells showed that those treated with ethanol, arsenite, lidocaine, and procaine also expressed nonspecific esterase activity, typical of monocytes, but did not adhere to plastic and had a cellular and nuclear morphology consistent with differentiation to metamyelocytes . Analysis of protein synthesis of HL-60 cells treated with 170 mM N-methylformamide, by the pulse labeling of cells for 2 h with {14C}leucine at various times, showed that the constitutive synthesis of both the Mr 90,000 and 70,000 heat shock proteins fell substantially after 2 h of exposure to N-methylformamide . When HL-60 cells were incubated with 1 M N-methylformamide, a toxic concentration of this agent, or were heat shocked, the synthesis of both the Mr 70,000 and Mr 90,000 proteins was induced . We propose that changes in heat shock protein synthesis may be an important element of the induction of differentiation of HL-60 cells, particularly as these proteins have recently been shown to regulate the stability of oncogene proteins, such as myc (Luscher, B., and Eisenman, R . N., Mol . Cell Biol., 8: 2504-2512, 1988).

Mycopathologia, 1988 Dec, 104(3), 171 - 80
Effect of antineoplastic agents and X-irradiation on the adherence of Candida spp . to human buccal epithelial cells in vitro; Ghannoum MA et al.; The role of chemotherapy, X-irradiation and a combination of both on the phenomenon of adherence of yeast to buccal epithelial cells (BEC) was investigated in vitro . Growth of three Candida spp . in the presence of eight of eleven antineoplastic agents led to reduction of adherence of the isolates tested (reduction between 30% and 61% of the control value), and this effect was observed whether exponential or stationary phase Candida cells were used . Exposure of C . albicans to various doses of radiation also led to a reduction in adherence of this yeast to BEC between 31% and 53% of the control value . This reduction was shown to be dose related . Similar results were obtained when BEC were exposed to radiation, and the effects of radiation treatment was accentuated when both yeast and BEC were irradiated simultaneously . Furthermore, treating C . albicans with a combination of chemotherapy and radiation led to the greatest reduction in adherence of yeast to BEC compared to when the yeast was treated with either chemotherapy or radiation alone (reduction between 63% to 74% as compared with control) . The possible mechanism/s involved in reduction of adherence of yeast to BEC are discussed.

Immunology, 1988 Dec, 65(4), 511 - 4
Monoclonal antibodies that recognize distinct epitopes of the macrophage type three complement receptor differ in their ability to inhibit binding of Leishmania promastigotes harvested at different phases of their growth cycle; Cooper A et al.; The macrophage receptor CR3 has been shown by several investigators to be involved in the binding of Leishmania promastigotes to host macrophages . This receptor is known to recognize iC3b and to mediate direct lectin-like attachment of particles such as yeast zymosan . In the present study, two anti-CR3 monoclonal antibodies, M1/70 and 5C6, which ligate different epitopes of murine CR3, have been used in conjunction with sodium salicyl hydroxamate (Saha; inhibits covalent ester linkages of C3 to an activator surface) to block binding of L . donovani and L . major promastigotes harvested at different phases of their growth cycle . M1/70 inhibited all promastigote binding . 5C6 and Saha blocked in parallel only the binding of peanut agglutinin (PNA)-positive late log and early stationary phase parasites . These results suggest that the binding PNA-positive parasites to CR3 is iC3b-mediated, while entry of the more infective PNA-negative late stationary phase promastigotes into host macrophages may involve direct lectin-like binding to CR3.

J Cell Biol, 1988 Dec, 107(6 Pt 2), 2669 - 78
Identification and localization of a novel, cytoskeletal, centrosome-associated protein in PtK2 cells; Baron AT et al.; Antisera raised against centrin (Salisbury, J.L., A.T . Baron, B . Surek, and M . Melkonian . 1984 . J . Cell Biol . 99:962-970) have been used, here, to identify a centrosome-associated protein with an Mr of 165,000 . Immunocytochemistry indicates that this protein is a component of pericentriolar satellites, basal feet, and pericentriolar matrix of interphase cells . These components of pericentriolar material are, in part, composed of 3-8-nm-diam filaments, which interconnect to form a three-dimensional pericentriolar lattice . We conclude that the 165,000-Mr protein is immunologically related to centrin, and that it is a component of a novel centrosome-associated cytoskeletal filament system . Microtubule organizing centers such as the flagellar apparatus of algal cells, spindle pole body of yeast cells, and centrosome of mammalian cells are homologous structures essential for cytoplasmic organization and cellular proliferation . Molecular cloning studies have recently shown that the cell cycle gene product CDC31, required for spindle pole body duplication, shares 50% sequence homology with centrin (Huang, B., A . Mengersen, and V.D . Lee . 1988 . J . Cell Biol . 107:133-140) . The evolutionary conservation of centrin-related sequences and immunologic epitopes to microtubule organizing centers of divergent phylogeny suggests that a functional attribute(s) may have been conserved as well . Elucidation of a common thread between these related molecules may be fundamental to our understanding of cell structure and function.

Pain, 1988 Dec, 35(3), 313 - 26
Enhancement of dynorphin gene expression in spinal cord following experimental inflammation: stimulus specificity, behavioral parameters and opioid receptor binding; Iadarola MJ et al.; The stimulus specificity for enhancement of dynorphin gene expression in rat spinal cord was studied by combined measurements of the peptide dynorphin A 1-8 and preprodynorphin mRNA levels during peripheral inflammation induced by several agents . The density of kappa receptors, the putative receptor for dynorphin peptides, was examined using receptor binding with autoradiographic visualization . Mu and delta receptor classes were also studied . All inflammatory agents tested (carrageenan, phorbol ester, yeast and Freund's adjuvant) rapidly induced edema and thermal hyperalgesia . All agents also induced a rapid (within 8 h) elevation in dynorphin mRNA and, in comparison, a delayed (within 2 days) elevation of dynorphin A 1-8 peptide; peak peptide levels were reached at 4 days . No alteration of kappa, mu or delta receptor binding was observed at 4 h or 4 days post inflammation . The rapid development of thermal hyperalgesia and elevation of dynorphin mRNA and peptide content indicates that the involvement of dynorphin-containing neurons in nociceptive processing does not require a chronic abnormality and a dynamic picture of opioid modulation of sensory processing emerges . These data also demonstrate that activation of dynorphin biosynthesis in spinal cord is a feature common to hyperalgesia and peripheral inflammation and is not restricted to any one type of inflammatory agent . The lack of alteration in receptors suggests that the physiological effects of an increased biosynthesis are not accompanied by a concurrent down-regulation of opiate receptors.

Eur J Obstet Gynecol Reprod Biol, 1988 Dec, 29(4), 305 - 13
A comparison of fluconazole and ketoconazole in the oral treatment of vaginal candidiasis; report of a double-blind multicentre trial; Kutzer E et al.; One hundred and eighty three patients with vaginal candidiasis were randomly allocated to treatment with either fluconazole in a single oral 150 mg dose, or ketoconazole, 200 mg twice daily for 5 days . Favourable clinical responses were obtained in 92% of the patients in the fluconazole group and in 89% of those in the ketoconazole group after 5-16 days . Long-term evaluation at 27-62 days showed favourable clinical responses in 86 and 88%, respectively . Candida was eradicated from the vagina in 77% of both treatment groups at the long-term evaluation . The relapse or reinfection rate was similar for both groups, ranging between 4-8% . One hundred and thirty two of the 160 patients who had rectal swabs cultured for yeast at baseline, gave positive results . Of the 27 patients in the fluconazole group whose rectal cultures remained negative at the long-term evaluation, 26 maintained mycological cure of their vaginal candidiasis . In contrast, patients with positive rectal cultures at this time were much less frequently associated with mycological cure . The results were similar for the patients in the ketoconazole group . Treatment-related side effects in both groups were few and minor . This double-blind multicentre study showed that a single oral 150 mg dose of fluconazole was as effective as 5 days of oral ketoconazole medication in the treatment of vaginal candidiasis.

J Leukoc Biol, 1988 Dec, 44(6), 500 - 7
Characterization of murine bronchoalveolar macrophage respiratory burst: comparison of soluble and particulate stimuli; Sugar AM et al.; Stimulation of the respiratory burst of murine bronchoalveolar macrophages obtained by lung lavage was studied using four different stimuli and different assay conditions . One soluble stimulus, phorbol myristate acetate (PMA), two intracellular particles, zymosan and Blastomyces dermatitidis conida, and one extracellular particle, B . dermatitidis yeast, were incubated with either freshly obtained macrophages in suspension or 2- and 48-hour macrophage monolayers . Suspension cultures were incubated with stimuli for 90 minutes and monolayers for 10 minutes before O2- was assayed . PMA did not elicit O2- production in macrophage suspensions or 2-hour macrophage monolayers, but 48-hour macrophage monolayers exhibited a 13-fold increase above control values (P = .0001) . On the other hand, zymosan elicited an increase in O2- production in both suspensions and monolayers, although monolayers incubated for 48 hours produced almost fourfold more O2- than the other systems (P = .025) . Opsonization had no effect on the ability of zymosan to elicit respiratory burst . B . dermatitidis conidia resulted in a two- to threefold increase in O2- production in macrophage suspensions and a five- to eightfold increase in 48-hour monolayers, representing significantly less respiratory burst stimulation than with either zymosan or PMA . Similarly, B . dermatitidis yeasts demonstrated similar submaximal stimulation of O2-, 3-4 times that over control values, and again this was less than zymosan and PMA . We conclude that 1) freshly obtained murine bronchoalveolar macrophages do not respond to PMA with an increase in O2- production, but that responsiveness is evident after 48 hours of incubation in monolayers; 2) B . dermatitidis conidia and yeasts do not stimulate respiratory burst activity to the same degree as zymosan or PMA; and 3) opsonization of zymosan is not necessary for stimulation of the murine bronchoalveolar macrophage oxidative burst, confirming previous data that functional complement receptors are not present on these cells.

Zh Mikrobiol Epidemiol Immunobiol, 1988 Dec, (12), 104 - 6
{The action of interferon preparations on strains of Legionella of various serogroups and species}; Tartakovskii IS et al.; The action of different preparations of interferon on Legionella strains has been studied in vivo and in vitro . The preparations of leukinferon at a concentration of 500 international units (I.U.) and reaferon at a concentration of 10,000 I.U . have been found to produce an inhibiting effect on Legionella strains in vitro, in a medium with carbon-yeast agar . Leukinferon at a concentration of 125 I.U . suppresses the growth of L . pneumophila also in a liquid medium . The preparation of leukinferon at a minimal concentration of 100 I.U . has been found to suppress the development of lethal infection in chick embryos infected with L . pneumophila strain Philadelphia 1.

Eur J Immunol, 1988 Dec, 18(12), 1951 - 7
Multiple T helper cell epitopes of the circumsporozoite protein of Plasmodium berghei; Romero PJ et al.; The present findings establish the lack of genetic restriction of the humoral immune response to sporozoites of Plasmodium berghei, corraborating earlier observations that mice of different strains can be protected by immunization with irradiated sporozoites . Most, if not all, anti-sporozoite antibodies are directed against the repetitive B cell epitope of the circumsporozoite (CS) protein . However, neither a peptide containing a dimer of this repeat (17.1), nor a peptide polymer containing multiple repeats induced an antibody response in mice of different H-2 and different genetic backgrounds . A yeast-derived recombinant, containing the repeat domain and part of the surrounding amino and carboxy-terminal regions of the P . berghei CS protein, induces very different levels of antibody in mice of diverse H-2 haplotypes . H-2j mice are high responders and the immunized mice are extensively protected against sporozoite challenge . The lymph node cells of the H-2j mice (but not from other strains) proliferated in the presence of peptide N, contained in the amino terminal region of the CS recombinant . Additional H-2-restricted T cell epitopes have been identified in amino and carboxy-terminal regions of the CS protein, and mice of most of the strains recognized multiple T cell epitopes . Two peptides representing T cell epitopes were synthesized in tandem with a peptide representing the B cell epitope, and were assayed for T helper activity in vivo . The antibody response of mice, primed by a single injection of sporozoites, was boosted very effectively by the administration of peptide N + 17.1 or peptide B-4 + 17.1 . The B-4 T cell epitope is located in the carboxy-terminal region of the CS protein and is recognized by mice of at least four different H-2 haplotypes . These observations demonstrate that the immune response to the CS protein of P . berghei is not genetically restricted and that it contains several T cell epitopes, some of which can function as helper epitopes . In addition, they show that a synthetic sporozoite vaccine can boost the immune response to sporozoites.

JAMA, 1988 Nov 25, 260(20), 3035 - 41
RNA splicing and genes; Sharp PA; The splicing of long transcripts of RNA (copied from DNA in the cell nucleus) into smaller, specific mRNA (ready for export to the protein-producing machinery in the cytoplasm) is an important event in the regulation of gene expression in eukaryotic cells . The splicing reaction occurs as a late step in the nuclear pathway for synthesis of mRNAs . This pathway commences with initiation of transcription by RNA polymerase II and probably involves an integrated series of steps each dependent on previous events . Splicing of precursors to mRNAs involves the formation of a spliceosome complex containing the 5' and 3' splice sites . This complex contains the evolutionarily highly conserved small nuclear RNAs (snRNAs) U2, U4, U5, and U6 . The most abundant snRNA, U1, is required to form the spliceosome and may be a part of the spliceosome . Analogues of these snRNAs have been identified in yeast . Assembly of the spliceosome probably involves the binding of a multi-snRNA complex containing U4, U5, and U6 snRNAs . Several observations suggest that the association of snRNAs in such complexes is quite dynamic . It is argued that the snRNAs in the spliceosome form a catalytic RNA structure that is responsible for the cleavage and ligation steps during splicing.

Cell, 1988 Nov 18, 55(4), 731 - 8
Site selection by the tRNA splicing endonuclease of Xenopus laevis; Mattoccia E et al.; To investigate the mechanism by which the purified Xenopus tRNA splicing endonuclease recognizes its splice sites, we utilized yeast pre-tRNA(3Leu) and pre-tRNA(Phe) variants constructed by in vitro mutagenesis . We found that the endonuclease interacts with conserved features of the mature tRNA domain . In particular, U8 and C56 may be examples of contact points between protein and RNA . Given that there are no conserved sequences at the splice junctions, the specificity of cutting at both splice sites is determined by the length of the anticodon stem . Although in general, the sequence of the intron is unimportant for splicing, there are some structural requirements.

Biochim Biophys Acta, 1988 Nov 16, 936(2), 187 - 98
Electrostatic interactions of 4-carboxy-2,6-dinitrophenyllysine-modified cytochromes c with physiological and non-physiological redox partners; Rush JD et al.; An analysis of the effect of electrostatic properties of 4-carboxy-2,6-dinitrophenyllysine (CDNP-lysine) cytochromes c on their reactions with strongly and weakly binding redox partners is given . For strongly binding systems (cytochrome-c oxidase, cytochrome-c reductase, sulphite oxidase and yeast cytochrome-c peroxidase) the magnitude of the dipole moments of the CDNP cytochromes c determines their relative reactivities . For weakly binding redox agents, such as hexacyanoferrate(III), cobalt(III)tris(1,10-phenanthroline), azurin and plastocyanin, the electrostatic potential at the haem edge accounts for the greater part of the relative activities . Relative rate data were obtained from the literature . It is concluded that the dipole moment of native cytochromes c may account for an approx . 50-fold increase in the efficiency of its physiological activity towards membrane-bound enzymes . A correction on a formula to describe the contribution of a molecular dipole moment to the ionic strength dependence of a bimolecular rate constant (Koppenol, W . H . (1980) Biophys . J . 29, 493-508) leads to an equation nearly identical to that obtained by Van Leeuwen et al . (Van Leeuwen, J.W., Mofers, F.J.M . and Verrman, E.C.I . (1981) Biochim . Biophys . Acta 635, 434-439).

Nucleic Acids Res, 1988 Nov 11, 16(21), 9995 - 10011
Specific and ubiquitous expression of different Zn finger protein genes in the mouse; Chowdhury K et al.; Zinc finger proteins (Zfp) are members of a multigene family encoding Zn mediated nucleic acid binding proteins . They have been isolated from various organisms including yeast, Drosophila, Xenopus mouse and human . All Zfp share the 28-30 amino acid long finger repeats containing conserved residues at specific positions . Some of these proteins have been identified as transcriptional regulatory factors . In this paper, we describe the isolation, DNA sequence determination and the expression pattern in developing embryos and adult tissues of 3 new members of the mouse Zfp . All of them are expressed as multiple transcripts . Unaltered level of mkr5 expression could be detected in 10-15 day whole embryo RNAs but its level started to decrease from day 16 . In the adult animal, predominant expression was detected only in the ovary . In contrast, mkr3 and 4 were expressed at a constant level in all embryos and tissues tested . These data suggest the presence of both tissue specific and ubiquitious Zfp in the mouse.

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