J Biol Chem, 1988 Nov 5, 263(31), 16499 - 503 Distinct and additive effects of elastase and endotoxin on expression of alpha 1 proteinase inhibitor in mononuclear phagocytes; Perlmutter DH et al.; Expression of alpha 1 proteinase inhibitor (alpha 1-PI) in human mononuclear phagocytes may provide a local mechanism for inactivation of serine proteases at sites of tissue injury, thereby preventing incidental damage to surrounding tissue and allowing for orderly initiation of repair . We have previously shown that serine (neutrophilic or pancreatic) elastase and lipopolysaccharide (LPS) each mediate an increase in the expression of alpha 1-PI in human peripheral blood monocytes and bronchoalveolar macrophages . In this study we demonstrate that elastase and LPS have an additive positive regulatory effect on alpha 1-PI expression . Distinct pretranslational and translational mechanisms of action for elastase and LPS, respectively, account for the additive effect . The possibility that translational regulation of alpha 1-PI by LPS involves a mechanism analogous to that of the yeast gene GCN4 during amino acid starvation and that of the human ferritin gene in response to iron is discussed. Cell, 1988 Nov 4, 55(3), 443 - 6 Converting a eukaryotic transcriptional inhibitor into an activator; Ma J et al.; GAL80, an inhibitor of the yeast transcriptional activator GAL4, is converted into an activator by inserting an acidic activating sequence into it . This hybrid activator does not bind to DNA directly, but is brought to DNA by interacting with a derivative of GAL4 that interacts with both DNA and GAL80. Z Lebensm Unters Forsch, 1988 Nov, 187(5), 436 - 9 Growth and aflatoxin production by Aspergillus parasiticus in a medium at different pH values and with or without pimaricin; Rusul G et al.; A glucose-yeast extract-salt medium containing 0, 5, 7.5, 10, 15 or 20 micrograms pimaricin/ml with an initial pH of 3.5 or 5.5 was inoculated with Aspergillus parasiticus WB 108 and incubated at 15 degrees or 28 degrees C . The pH, weight of mycelium and amount of aflatoxin produced were determined after 3, 7, and 10 days and after 14, 21, and 30 days when incubation was at 28 degrees or 15 degrees C, respectively . Increasing the concentration of pimaricin in the medium with an initial pH of 5.5 decreased the amounts of aflatoxin B1 and G1 produced after 3 days of incubation . When the initial pH of the medium was 3.5, no growth or toxin production occurred after 3 days of incubation in the medium containing 7.5 micrograms or more of pimaricin/ml . The presence of 20 micrograms of pimaricin/ml inhibited growth and toxin production after 7 days of incubation . When cultures were incubated at 15 degrees C, there was a lag phase which extended from 9 to 16 days, and the amounts of aflatoxin produced decreased with an increasing concentration of pimaricin . Pimaricin did not completely inhibit the growth and aflatoxin production by A . parasiticus . Pimaricin, in combination with a low pH, low temperature or 4% or 6% NaCl, initially caused slow mycelial growth and low toxin production, but the mold overcame the inhibitory effects and produced substantial amounts of mycelium and toxin. J Burn Care Rehabil, 1988 Nov-Dec, 9(6), 599 - 601 Noncandidal, fungal infections of the burn wound; Burdge JJ et al.; Candidal (yeast) and noncandidal (filamentous fungal) wound infections have become an increasingly important cause of burn-associated morbidity and mortality . However, these two diseases differ markedly in their epidemiology, onset, appearance, diagnosis, and treatment . In the last four years, we have had five cases of noncandidal sepsis (four mucoraceae and one aspergillus) that are illustrative of these differences . Early recognition and diagnosis are essential . Radical excision when the infection is confined to superficial invasion of only one anatomic area is key to decreasing mortality from this lethal disease. Mutat Res, 1988 Nov-Dec, 209(3-4), 135 - 40 The clastogenic activity of 1,6-dinitropyrene in peripheral human lymphocytes; Adams K et al.; The ability of 1,6-dinitropyrene to induce chromosome damage in peripheral human lymphocyte cultures has been demonstrated . Low levels of clastogenic activity were detected following 3-h treatments with 1,6-dinitropyrene in the presence of a rat-liver cytosol fraction . The clastogenic activity reached a peak at a concentration of 1.25 micrograms/ml of 1,6-dinitropyrene after which the frequency of aberrations decreased . This unusual genotoxic dose response is similar to that found previously in yeast and rat-liver cells . The fact that a positive result was obtained using human lymphocytes shows that, in the presence of the appropriate activation system, dinitropyrene is genotoxic in human cells. Mycopathologia, 1988 Nov, 104(2), 75 - 9 Comparison of some screening methods for aflatoxigenic moulds; Abarca ML et al.; Thirty seven strains of the Aspergillus flavus group isolated from animal mixed feeds have been screened for their ability to produce aflatoxins in yeast extract and sucrose (YES), aflatoxin producing ability (APA), and coconut agar medium (CAM) media . The concentration and detection of the aflatoxins by different methods is compared . Five known aflatoxin-positive and one aflatoxin-negative strains have been used as controls . Only 5 out of the 37 strains (13.5%) were aflatoxin-producers in YES medium . Of these five strains and the five known aflatoxin-positive strains, only three showed blue fluorescence in APA medium and four in CAM medium . Generally, the aflatoxin concentration in CAM medium was higher than in YES and APA media . Using the 'agar-plug method' and by direct spotting of the YES broth on TLC plates, some aflatoxin-producing strains were not detected. Proc Natl Acad Sci U S A, 1988 Nov, 85(21), 8131 - 5 Dilute suppressor dsu acts semidominantly to suppress the coat color phenotype of a deletion mutation, dl20J, of the murine dilute locus; Moore KJ et al.; The murine dilute suppressor (dsu) gene is the only unlinked trans-acting suppressor identified in mammals . dsu, which was originally reported to be recessive, was recognized by its ability to suppress the coat color phenotype of a retroviral insertion mutation, dv, of the murine dilute (d) locus . This insertion mutation resulted from the integration of an ecotropic murine leukemia virus into noncoding sequences of the dilute gene . Therefore, dsu may act like other allele-specific recessive suppressors identified in Drosophila melanogaster and yeast that suppress mutations induced by retrotransposon insertions . To investigate this possibility, we have examined whether dsu could suppress a spontaneously arising allele of d, dl20J, which is shown here to result from a 3.5-kilobase deletion . These studies indicate that dsu does not function like other eukaryotic suppressor genes that suppress retrotransposon-induced mutations . We also show that dsu is not, as originally reported, a recessive gene but is semidominantly inherited . Collectively, these results allow us to propose a mechanism for the suppressor activity of dsu. Pediatr Dermatol, 1988 Nov, 5(4), 260 - 2 Pityriasis (tinea) versicolor in infancy; Nanda A et al.; Pityriasis (tinea) versicolor is a common disorder of adults . We cared for five infants (four males and one female) with the disease . Diagnosis was confirmed by potassium hydroxide preparation demonstrating the filaments of Malassezia furfur and/or Pityrosporum orbiculare, the yeast form . Three patients had lesions in the neonatal period . The mother of one baby had pityriasis versicolor . Two patients were siblings . One baby had associated atopic dermatitis and two had a positive family history of atopy. Laryngoscope, 1988 Nov, 98(11), 1173 - 7 In vitro effectiveness of 13 agents in otomycosis and review of the literature; Stern JC et al.; Many agents have been recommended for treating otomycosis, but no preparation has been widely accepted . To compare the effectiveness of many recommended preparations, we performed an in vitro study using 15 species of fungi and yeast cultured from patients presenting with otomycosis during the past year . By measuring zones of inhibition, we assessed the effectiveness of aqueous Merthiolate, Burow's solution (2%), VoSol HC, VoSol plain, Cortisporin suspension, clotrimazole 1%, Mycostatin, amphotericin B, ethanol 95%, miconazole, tolnaftate 1%, natamycin, and flucytosine . Most otic preparations showed little or no growth inhibition . However, Merthiolate was very effective against all organisms tested, clotrimazole was very effective against most yeast and fungi tested, and nystatin had the widest spectrum of activity among the antifungals . Tolnaftate was ineffective . Vigorous cleaning of the external auditory canal remains the mainstay in treating otymycosis, but proper laboratory identification and suitable topical therapy are also important in dealing with this capricious infection. Biochem Cell Biol, 1988 Nov, 66(11), 1169 - 76 Effect of microenvironment and cell-line type on carbohydrate-binding proteins of macrophage-like cells; Gabius HJ et al.; The pattern of sugar inhibition of rosette formation, a model for intercellular interaction between cultured cells and glutaraldehyde-fixed, trypsinated rabbit erythrocytes, served to infer the presence of carbohydrate-binding proteins . This profile from cell extracts for the two murine macrophage-like cell lines, P388D1 and J774A.1, was comparatively analyzed by affinity chromatography on supports with immobilized carbohydrates (lactose, L-fucose, N-acetyl-D-glucosamine, N-acetyl-D-galactosamine, and maltose) or with the immobilized mannose-rich yeast glycoprotein mannan or fetuin-derived glycopeptides containing sialic acid residues . After elution with specific sugar in the absence of Ca2+ ions, the proteins were separated by sodium dodecyl sulfate - polyacrylamide slab gel electrophoresis . The composition of carbohydrate-binding proteins of the two lines clearly exhibited quantitative and qualitative differences . Moreover, the pattern of P388D1 cells was also demonstrated to change significantly in response to alterations in the conditions of the physiological environment . These alterations were imposed by in vitro growth, by subsequent in vivo growth in nude mice, and by re-adaptation of cells to culture after in vivo passage . Collectively, our observations and other physiological and biochemical reports on macrophage lectins indicate that the presence of sugar receptors with different specificities may be an indicator of macrophage differentiation, being reversibly modulated to a considerable extent by external factors, e.g., microenvironment . Extensive but selective alterations in this respect could play an important role in the control of recognition and effector mechanisms within diverse functions of macrophage subpopulations. J Assoc Off Anal Chem, 1988 Nov-Dec, 71(6), 1158 - 61 Liquid chromatographic determination of L-ascorbate 2-polyphosphate in fish feeds by enzymatic release of L-ascorbate; Wang XY et al.; An accurate method was devised to assay L-ascorbic 2-polyphosphate esters (AsPP) in fish feed by phosphatase digestion followed by determination of the released L-ascorbic acid (AsA) . Compressed yeast and dithiothreitol are added to the phosphatase reaction mixture to give 95-100% recovery of AsA, which is quantitated by reverse-phase liquid chromatography (LC) with electrochemical detection . Chromatograms of all feed digests showed baseline resolution of AsA . In 3 feeds, to which 75-125 ppm AsA equivalents in the form of AsPP were added, the assay procedure gave 98-100% recovery of AsA. Genes Dev, 1988 Nov, 2(11), 1353 - 63 Homologous integration in mammalian cells without target gene selection; Jasin M et al.; Homologous integrations into a nonselectable target locus have been highly enriched for following DNA transfections into mammalian cells . The target gene, the SV40 early region in COS1 cells, provides transcription signals to activate a defective selectable marker, the gpt gene . We find that nearly half of the selected clones have integrated the gpt gene at the homologous sequence in the COS1 genome . This is an estimated 100-fold enrichment for homologous events compared with transfections in which the gpt gene is transcriptionally active . As shown for yeast integration events, a double-strand break at a position of homology between the transfected DNA and the genomic target is necessary to achieve a high frequency of homologous integrations . Furthermore, the arrangement of sequences at the integration site includes a repair of the double-strand gap, which was present on the transfected DNA, suggesting that similarities exist between yeast and mammalian integrations . The experimental design, in which a defective marker is activated following a homologous integration, may have general applications for gene targeting in mammalian cells. Chest, 1988 Nov, 94(5), 949 - 53 Open lung biopsy diagnosis of diffuse pulmonary infiltrates after marrow transplantation; Crawford SW et al.; The results were reviewed of 111 open lung biopsies (OLB) performed on 109 marrow transplantation recipients with diffuse pulmonary infiltrates between January 1983 and July 1987 . We determined the frequency and types of infections identified, and the relationship to time after transplantation . Infection was found in 70 of the 111 cases (63 percent) and cytomegalovirus (CMV) was present in 90 percent of all cases with infection . Infection was identified in only five of 26 (19 percent) cases within the first 30 days after transplant, and when present, was viral . The prevalence of infection after 30 days (over 75 percent of 85 cases) was significantly higher (chi 2 = 26.2, p = 0.00001) . Bacterial or yeast infections were found in only four cases (4 percent) (two cases each), and Pneumocystis carinii in six cases (6 percent) . Simultaneous infection with two or more organisms was found in four cases (4 percent) . Four of 25 autopsies performed within ten days after OLB revealed fungal infections with Aspergillus not detected at OLB . Thus, the prevalence of infection detected by OLB is low within the first 30 days after marrow transplantation among patients receiving broad spectrum antibiotics . CMV infection is found in most transplantation recipients who undergo OLB with diffuse infiltrates between days 30 and 180. J Immunol, 1988 Nov 1, 141(9), 3170 - 6 Perturbation of beta-glucan receptors on human neutrophils initiates phagocytosis and leukotriene B4 production; Czop JK et al.; Normal human neutrophils were stimulated with the yeast cell wall product, zymosan, and examined for two biologic responses, ingestion of particles and production of leukotriene B4 (LTB4), under conditions that were comparable and optimal for the quantitation of each response . Monolayers of adherent neutrophils ingested unopsonized zymosan particles, at particle-to-cell ratios of 12.5:1 to 125:1, in a dose- and time-related manner . At a ratio of 125:1, the percentages of neutrophils ingesting greater than or equal to 1 and greater than or equal to 3 zymosan particles reached plateau levels of 55 +/- 6 and 32 +/- 9% (mean +/- SD, n = 8), respectively, within 30 min . At this same ratio, neutrophils during gravity sedimentation with zymosan particles synthesized LTB4 in a time-dependent manner for at least 45 min . The maximum amount of immunoreactive LTB4 released into supernatants was 3.8 +/- 1.2 ng per 10(6) neutrophils (mean +/- SD, n = 5) and the corresponding total immunoreactive LTB4 was 6.2 +/- 1.9 ng per 10(6) neutrophils . Treatment of 2 x 10(7) suspended neutrophils with 250 micrograms of trypsin for 20 min before concurrent assessment of neutrophil phagocytosis and LTB4 production reduced both of these responses by about 50% . Pretreatment of neutrophils with 800 micrograms/ml of soluble yeast beta-glucan inhibited their ingestion of zymosan by 84% (mean +/- SD, n = 3), with 50% inhibition occurring with 100 micrograms/ml of soluble beta-glucan; 800 micrograms/ml of soluble yeast alpha-mannan had no inhibitory effect . Pretreatment of neutrophils with 400 micrograms/ml of soluble yeast beta-glucan inhibited neutrophil synthesis of LTB4 by 90%, with 50% occurring with 200 micrograms/ml; 400 micrograms/ml of soluble yeast alpha-mannan had no inhibitory effect . The presence of 1.25 micrograms/ml of cytochalasin B during incubation with zymosan particles reduced neutrophil phagocytosis from 65 to 6%, and neutrophil synthesis of LTB4 from total levels of 6.0 +/- 0.3 ng/10(6) cells to zero (mean +/- SD, n = 3) . Pretreatment with either cytochalasin B or vinblastine did not alter neutrophil generation of LTB4 induced by calcium ionophore . Neutrophils pretreated with vinblastine, at 4 x 10(-6) to 4 x 10(-4) M, and then maintained at one-half these concentrations during incubation with unopsonized zymosan particles exhibited no diminution in particle ingestion, but were markedly reduced in zymosan-induced synthesis of LTB4.(ABSTRACT TRUNCATED AT 400 WORDS) Stain Technol, 1988 Nov, 63(6), 335 - 8 A method for increasing the number of mitoses available for cytogenetic analysis in rainbow trout; Lozano R et al.; A new method for the cytogenetic analysis of fishes with special interest for the rainbow trout (Salmo gairdneri Rich.) is described . This reliable method that includes treatment with a yeast solution provides high quality spreads of a great number of metaphases. Mol Cell Biol, 1988 Nov, 8(11), 4727 - 35 Structure and expression of ubiquitin genes of Drosophila melanogaster; Lee HS et al.; We isolated and characterized two related ubiquitin genes from Drosophila melanogaster, polyubiquitin and UB3-D . The polyubiquitin gene contained 18 repeats of the 228-base-pair monomeric ubiquitin-encoding unit arranged in tandem . This gene was localized to a minor heat shock puff site, 63F, and it encoded a constitutively expressed 4.4-kilobase polyubiquitin-encoding mRNA, whose level was induced threefold by heat shock . To investigate the pattern of expression of the polyubiquitin gene in developing animals, a polyubiquitin-lacZ fusion gene was introduced into the Drosophila genome by germ line transformation . The fusion gene was expressed at high levels in a tissue-general manner at all life stages assayed . The ubiquitin-encoding gene, UB3-D, consisted of one ubiquitin-encoding unit directly fused, in frame, to a nonhomologous tail sequence . The amino acid sequence of the tail portion of the protein had 65% positional identity with that of yeast UBI3 protein, including a region that contained a potential nucleic acid-binding motif . The Drosophila UB3-D gene hybridized to a 0.9-kilobase mRNA that was constitutively expressed, and in contrast to the polyubiquitin gene, it was not inducible by heat shock. Philos Trans R Soc Lond B Biol Sci, 1988 Oct 31, 321(1207), 327 - 48 Genetic factors as determinants of infectious disease transmission in human communities; Weatherall DJ et al.; Genetic factors may play an important role in individual susceptibility to infection . Hitherto this problem has been investigated by attempting to relate the distribution of genetic polymorphisms in populations to present or past infection, or by analysing specific infections by classical twin studies or group comparisons . There is reasonable evidence that the common red-cell polymorphisms involving haemoglobin, enzymes or membrane have been maintained by relative resistance to malaria . Blood-group heterogeneity, including secretor status, may reflect varying susceptibility to bacterial, virus and yeast infection . There is increasing evidence that the HLA-DR system may be involved in modifying the clinical course of bacterial, virus and parasitic infection . So far no specific resistance or susceptibility loci similar to those found in murine models have been found in man . DNA analysis, particularly involving restriction fragment length polymorphism associations with candidate genes, offers a valuable new approach to this problem. Biochem Biophys Res Commun, 1988 Oct 31, 156(2), 874 - 81 Molecular cloning and nucleotide sequence of murine 2,3-bisphosphoglycerate mutase cDNA; Le Boulch P et al.; Cloning and sequencing of a murine cDNA with the entire coding region of 2,3-bisphosphoglycerate mutase is reported, as a prerequisite for further expression studies of this erythroid specific enzyme in Friend mouse erythroleukemia cells . A comparison between species of the deduced amino acid sequences of these proteins shows 20 substitutions between mouse and human and 21 between mouse and rabbit: none of these substitutions are in positions assumed to be in the active site . Amino acid alignment with the other related enzymes, the phosphoglycerate mutases, in combination with crystallographic data from yeast phosphoglycerate mutase, gives some insight into the structure/function correlation for this protein family . Amino acid residues which are most likely critical for either 2,3-bisphosphoglycerate mutase or phosphoglycerate mutase function are pointed out . Concerning the phylogenetic analysis, phosphoglycerate mutases B and M from mammalians appear to have diverged with the yeast enzyme from a common ancestor, before the emergence of the 2,3-bisphosphoglycerate mutases. J Biol Chem, 1988 Oct 25, 263(30), 15785 - 90 Isolation and characterization of the human 2,3-bisphosphoglycerate mutase gene; Joulin V et al.; The human 2,3-bisphosphoglycerate mutase gene was isolated from genomic libraries and analyzed by Southern blots and DNA sequencing . The transcription initiation site was localized by primer extension as well as by S1 protection of the mRNA . The gene extends over 22 kilobase pairs; it is composed of two introns (8.8 and 11.5 kilobase pairs long) and three exons (84, 662, and 965 base pairs long) . The second exon correlates with a functional subdomain of the protein, as shown by comparison with the yeast phosphoglycerate mutase structure . The sequence TAGAAAA was found 30 bases upstream from the transcription initiation site and could be analogous to the TATA box . A sequence homologous to the CCAAT box was found twice, at positions -75 and -178 . There is no GC-rich sequence or GC box in the 5'-flanking region of the gene . Northern blot analysis indicates that the 2,3-bisphosphoglycerate mutase mRNA is detected mainly in erythroid tissues and cell lines, although it is also present in low amounts in a nonerythroid tissue . A comparison of the 5'-upstream sequences with other promoters active only in erythroid cells did not reveal any common signal that could be responsible for the "erythroid promoter." Cell, 1988 Oct 21, 55(2), 371 - 8 cdc2 is a component of the M phase-specific histone H1 kinase: evidence for identity with MPF; Arion D 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 . In starfish oocytes, the hormone 1-methyladenine triggers synchronous meiotic divisions that are accompanied by a rapid 30-fold stimulation of H1K activity . We have substantially purified this activated enzyme and find that it is enriched for a protein of 34 kd . Quantitative immunoblotting of the column fractions with antibodies raised against p34, the product of the fission yeast cdc2 gene, revealed complete coelution of the H1K activity and a 34 kd anti-cdc2 cross-reactive protein . Starfish H1K also displayed the same apparent molecular weight, on a molecular sizing column, as the mitotically activated p13/p34/p62 protein kinase complex of HeLa cells . p13, the product of the fission yeast suc1+ gene, interacts tightly with p34 in yeast, Xenopus, and HeLa cells . H1K from starfish binds strongly to p13-Sepharose and the time course of 1-methyladenine-induced H1K activation, whether assayed in crude extract or on p13-Sepharose beads, is identical . These results indicate that a cdc2 homolog is a subunit of the M phase-specific H1K of starfish meiotic oocytes . Since this protein is also a subunit of the M-phase promoting factor (MPF) of Xenopus oocytes, we suggest that H1K and MPF are the same entity, and that histone H1 is likely to be one substrate of the pleiotropic MPF. Eur J Biochem, 1988 Oct 15, 177(1), 1 - 7 Purification and characterization of the 90-kDa heat-shock protein from mammalian tissues; Yonezawa N et al.; The 90-kDa heat-shock protein (HSP90) has been purified from mammalian tissues, mouse liver and porcine brain, with a good yield by a new method involving hydrophobic chromatography . Mouse liver HSP90 and porcine brain HSP90 were compared with mouse lymphoma HSP90 which was purified from T lymphoma cell line, L5178Y, by a modification of the previously reported method . These three HSP90s were indistinguishable from one another in amino acid composition, one-dimensional peptide mapping, elution pattern of proteolytic fragments (trypsin- or V8-protease-cleaved) in reverse-phase high-performance liquid chromatography, reactivity with the antibody against mouse T lymphoma HSP90 and the ability to bind to F-actin . The amino acid sequences of three portions (total 47 amino acid residues) of lymphoma HSP90 were determined and they were homologous to those of the corresponding portions of Drosophila HSP83A and yeast HSP90 . These results suggest that HSP90 is a highly conserved protein during evolution. Cell, 1988 Oct 7, 55(1), 135 - 44 A human protein specific for the immunoglobulin octamer DNA motif contains a functional homeobox domain; Ko HS et al.; The homeobox domain is shared by Drosophila homeotic proteins, yeast mating type proteins, and some functionally uncharacterized mammalian proteins . A lymphoid-restricted human protein that binds to the immunoglobulin octamer regulatory motif was shown to contain an amino acid sequence that has 33% amino acid identity with the consensus sequence of the previously cloned homebox domains . This homeobox gene was localized to chromosome 19, thus mapping separately from other human homebox genes . A mutant protein containing amino acid substitutions within a putative helix-turn-helix motif in the homeobox domain did not bind DNA detectably . This human homeobox protein was shown to bind the same DNA sequence as the homeobox domains of the yeast mating type proteins and Drosophila homeotic protein, suggesting that homeobox proteins may have closely related DNA binding characteristics. Breast Cancer Res Treat, 1988 Oct, 12(2), 235 - 43 Hepatic clearance and metabolism in the rat of a human breast cancer associated glycoprotein (GCDFP-15); Toth CA et al.; Gross Cystic Disease Fluid Protein (GCDFP-15) is a 60,000 dalton glycoprotein isolated from human breast cyst fluid, composed of four 15,000 dalton monomers . Carbohydrate analysis indicates that each monomer has a single carbohydrate chain of the complex type . GCDFP-15 intravenously injected into rats showed a rapid circulatory clearance, the rate of clearance being faster in female animals {t1/2 = 12.8 (+/- 2.0) min . females, and 16.7 (+/- 2.6) min . males} . The major organs of clearance were the liver (70%) and kidneys (15%) . Immunoperoxidase staining showed localization in Kupffer cells and the proximal convoluted tubules of the kidney . Removal of sialic acid from GCDFP-15 resulted in a more rapid clearance (t1/2 = 2.2 min) by the liver (85%) . This clearance was inhibited by coinjection of asialo alpha 1 acid glycoprotein . About 3% of GCDFP-15 was excreted in bile with a transit time through the liver of 38 min . Examination of the uptake of GCDFP-15 by isolated rat Kupffer cells showed that yeast mannan, fucosylated BSA, and carcinoembryonic antigen (CEA) failed to inhibit uptake, though the binding of GCDFP-15 was clearly saturable . This suggests that a novel receptor system on the rat Kupffer cell may be responsible for GCDFP-15 clearance. Biochem Int, 1988 Oct, 17(4), 611 - 5 Antibodies against alcohol dehydrogenase and lactate dehydrogenase inhibit the activities of other unrelated NADH-requiring dehydrogenases; Srivastava A et al.; Polyclonal rabbit antibodies to NADH-requiring enzymes such as yeast alcohol-dehydrogenase (ADH) and lactate-dehydrogenase (LDH) immunoinhibit the activities of other unrelated dehydrogenases . The immunoinhibition of malate-dehydrogenase (MDH) activity by anti-yeast ADH IgG and anti-hog LDH IgG was dependent on the concentration of antibodies and time . This demonstration of cross-reactivity with unrelated enzyme proteins reveals the existence of an antigenic site around the NADH binding region in each of these enzymes . Pre-treatment of the enzyme with NADH resulted in complete protection against immuno-inactivation . The competitive binding of NADH and the ineffectiveness of ATP establish the difference in the antigenic site around the NADH- and ATP-binding region. Mol Gen Genet, 1988 Oct, 214(2), 348 - 52 Selection and characterization of ura5 mutants of Histoplasma capsulatum; Worsham PL et al.; The combined use of non-aggregating Histoplasma capsulatum strains and a defined medium which allows quantitative plating of the yeast phase has allowed us to select 5-fluoroorotic acid (5-FOA)-resistant mutants of this dimorphic fungus . Approximately two-thirds of the 5-FOA-resistant strains were auxotrophic for uracil; all were deficient in orotidine-5'-monophosphate pyrophosphorylase (OMPpase) activity . One class of OMPpase mutant (alpha), which retained a low level of OMPpase activity, was auxotrophic in the yeast phase (37 degrees C) but grew slowly in the mycelial phase (25 degrees C) without exogenous uracil . This phenotype was not due to a temperature-sensitive OMPpase activity . Both wild-type and alpha mutants had a higher OMPpase activity in the mycelial phase than the yeast phase; this increased activity may be sufficient to allow mycelial growth of alpha mutants. Clin Exp Rheumatol, 1988 Oct-Dec, 6(4), 355 - 9 Random locomotion and phagocytosis of monocytes in undiluted normal serum and in undiluted SLE serum; Rydgren L et al.; Time-lapse filming was performed with normal monocytes in normal serum and in SLE serum, with and without yeast cell phagocytosis . Undiluted sera were used . After incubation of the monocytes with SLE serum we found a decreased velocity of random movement and a change of morphology to more condensed, rounded-up and un-spread cells with a decreased yeast cell phagocytosis . No obvious correlation was found between the SLE serum induced impairment of monocyte function and disease activity, complement levels or the presence of immune complexes . We conclude that monocyte in vitro membrane functions, such as random motility, spreading and yeast cell phagocytosis, are impaired in the presence of undiluted SLE serum. Anal Biochem, 1988 Oct, 174(1), 46 - 53 The identification of furanose and pyranose ring forms of the reducing units of oligosaccharides; Pazur JH et al.; Several types of oligosaccharides with glucose, arabinose, or galactose units have been prepared by chemical degradation of oligosaccharides of known structures and by enzymatic syntheses utilizing macerans amylase or yeast galactosyltransferase and appropriate substrates and cosubstrates . The ring forms of reducing units of these oligosaccharides and of oligosaccharides composed of glucose and mannose have been identified by a combined method of analysis based on methylation, gas-liquid chromatography, and mass spectrometry . In the dimethyl sulfoxide solvent used for the methylations, the oligosaccharides with arabinose or galactose units at the reducing ends occur as arabinofuranose or galactofuranose ring forms to the extent of 55 and 65%, respectively . The remainder of these monosaccharide units are present in the pyranose ring form . Oligosaccharides with glucose or mannose units at the reducing ends occur primarily in the pyranose ring form . An interesting observation is the finding that the reducing units which carry substituents linked by alpha-glycosidic linkages occur in a higher percentage in the furanose ring forms than those which carry substituents linked by beta-glycosidic linkages. Immun Infekt, 1988 Oct, 16(5), 175 - 8 {Evaluation of the efficacy of a recombinant hepatitis B vaccine}; Polywka S et al.; A recombinant vaccine against hepatitis B derived from yeast cells (Gen-HB-Vax, Co . MSD/Behring) has been evaluated in 59 healthy young volunteers (37 men, 22 women) with an average age of 24.4 years . The seroconversion rate was 100%, and no major side effects were observed . During a follow-up period of 24 months concentrations of antibodies against HBsAg were shown to decline to one tenth within 16 to 17 months . Triple vaccination led to protective antibody titres for about 27 months on average . Based on these findings we suggest the following recommendations concerning revaccination: an anti-HBs titre of more than 10,000 mIU/ml four weeks after third vaccination should be reassessed 3-5 years later, and titres between 1000 and 10,000 mIU/ml after 2-3 years . A control of the antibody titre should be performed after 1-2 years if the titre is 200-1000 mIU/ml after the third vaccination, and after 6-12 months if it is 100-200 mIU/ml . Antibody titres between 10 and 100 mIU/ml should already be reassessed about 3-6 months later . We recommend an immediate revaccination for persons with anti-HBs titres below 10 mIU/ml . This corresponds to the course of antibody concentrations which could be seen in former studies with the conventional serum-derived vaccine . Maximum anti-HBs titres are slightly below those observed with the serum-derived vaccine. Eur J Cancer Clin Oncol, 1988 Oct, 24(10), 1655 - 9 Evaluation of a Candida antigen detection test (Cand-Tec) in the diagnosis of deep candidiasis in neutropenic patients; Piens MA et al.; The diagnostic efficiency of a serum Candida antigen detection test Cand-Tec test) was prospectively investigated in 104 leukemic patients treated by intensive chemotherapy or allogeneic bone marrow transplantation . Candida antigen titers were determined on admission and then weekly as long as patients remained neutropenic . Nine patients had a proven disseminated yeast infection (diagnosed only at autopsy in five cases) . The highest Candida antigen titers were 1:2 in two patients and 1:4 or more in seven patients (sensitivity: 76% for this last titer) . This highest titer was observed 12 days before to 3 days after the diagnosis . Seven out of the 97 patients without proven deep candidiasis had a maximum titer of 1:4 (specificity: 93%) . The positive predictive value was 50% for a titer of 1:4 and 24% for a titer of 1:2, whereas the negative predictive value was 100% for a titer of 1:4 and 97% for a titer of 1:2 . Patients with elevated titers were mostly treated by chemotherapy, were older and had a worse prognosis than those with negative titers, although the duration of neutropenia was similar . It is concluded that Candida antigen detection is a reliable method of diagnosis of deep candidiasis in neutropenic patients . The clinical interest in this test, with special regard to empiric antifungal therapy, is discussed. Oncogene, 1988 Oct, 3(4), 391 - 5 Colony stimulating factor-1 induced growth stimulation of v-fms transformed fibroblasts; Lyman SD et al.; The v-fms oncogene, which is capable of transforming fibroblasts, was derived by recombination of a feline leukemia virus with a cellular gene (c-fms) that encodes the receptor for colony stimulating factor 1 (CSF-1) . We examined the capacity of recombinant human CSF-1 (produced in a yeast expression system) to stimulate the growth of v-fms transformed rat fibroblasts . Recombinant human CSF-1 bound to v-fms transformed fibroblasts with high affinity (apparent Kd = 6.0 x 10(-10) M); only non-specific binding was observed on control cells . The number of colonies formed in soft agar by v-fms transformed cells was increased by CSF-1 treatment in a dose-dependent manner; a nine fold increase in the number of colonies was seen in the presence of 10(-8) M CSF-1 . CSF-1 did not stimulate the growth of either non-transformed cell lines, a non-transformed cell line that expresses a mutated v-fms protein on the cell surface, or cells transformed by the v-fgr oncogene . The growth stimulating effect of CSF-1 on v-fms transformed cells was also seen in monolayer culture . The v-fms transformed cells treated with CSF-1 had a more refractile, rounded morphology than non-treated cells; no morphology change was observed in CSF-1 treated control cells . CSF-1 treatment also increased both the number and size of foci that arose from fibroblasts following transfection with the v-fms oncogene . These data show that the altered CSF-1 receptor encoded by the v-fms oncogene retains a capacity to bind, and be stimulated by, human CSF-1. Genes Dev, 1988 Oct, 2(10), 1333 - 43 Analysis of the lethal interaction between the prune and Killer of prune mutations of Drosophila; Biggs J et al.; The third-chromosome mutation Killer of prune (K-pn) causes no phenotype by itself, but causes lethality in individuals homozygous for the nonlethal X-chromosome mutation prune (pn) . We have recovered 12 gamma-ray-induced revertants of Killer of prune . All of the revertants fail to complement a recessive cell lethal mutation in the abnormal wing discs (awd) gene . We present evidence that Killer of prune is a mutation in the awd gene . First, revertant awdKR14 leads to reduced accumulation of the awd gene product, but does not affect flanking genes . Second, when a copy of the awd gene is cloned from Killer of prune homozygous flies and injected into embryos, transformants express the lethal interaction with prune . In individuals of the genotype pn; awdK-pn/awd+ the awd mRNA is present at normal levels but the awd polypeptide fails to accumulate . The absence of the awd gene product in such individuals is the cause of death . Although the awd polypeptide is a subunit of a cytoplasmic protein, its sequence is similar to subunit V of yeast cytochrome oxidase. Proc Natl Acad Sci U S A, 1988 Oct, 85(19), 7177 - 81 Cloning and sequencing of cDNA encoding human DNA topoisomerase II and localization of the gene to chromosome region 17q21-22; Tsai-Pflugfelder M et al.; Two overlapping cDNA clones encoding human DNA topoisomerase II were identified by two independent methods . In one, a human cDNA library in phage lambda was screened by hybridization with a mixed oligonucleotide probe encoding a stretch of seven amino acids found in yeast and Drosophila DNA topoisomerase II; in the other, a different human cDNA library in a lambda gt11 expression vector was screened for the expression of antigenic determinants that are recognized by rabbit antibodies specific to human DNA topoisomerase II . The entire coding sequences of the human DNA topoisomerase II gene were determined from these and several additional clones, identified through the use of the cloned human TOP2 gene sequences as probes . Hybridization between the cloned sequences and mRNA and genomic DNA indicates that the human enzyme is encoded by a single-copy gene . The location of the gene was mapped to chromosome 17q21-22 by in situ hybridization of a cloned fragment to metaphase chromosomes and by hybridization analysis with a panel of mouse-human hybrid cell lines, each retaining a subset of human chromosomes. Biochem Biophys Res Commun, 1988 Sep 30, 155(3), 1278 - 83 The nucleolar protein, B-36, contains a glycine and dimethylarginine-rich sequence conserved in several other nuclear RNA-binding proteins; Christensen ME et al.; The amino terminal sequence of the 34 kD nucleolar protein B-36 isolated from the slime mold Physarum polycephalum has been determined . This portion of B-36 is rich in glycine, phenylalanine and the modified amino acid asymmetrical dimethylarginine (DMA) and is 65% identical to that for fibrillarin, a similar and potentially homologous 34 kD nucleolar protein from rat . The terminus of B-36 contains an interesting nine amino acid sequence, Gly-DMA-Gly-Gly-Phe-Gly-Gly-DMA-Gly, which is precisely repeated three times in the 110 kD nucleolar protein nucleolin . Similar sequences have also been reported in a yeast nucleolar protein (SSB-1) and several hnRNP proteins (rat A1 and brine shrimp GRP33) . The conserved nature of this unusual sequence is suggestive of an important function which may include RNA-binding since several of these proteins share this feature. JAMA, 1988 Sep 23-30, 260(12), 1734 - 8 Lack of response to recombinant hepatitis B vaccine in nonresponders to the plasma vaccine; Weissman JY et al.; Yeast recombinant hepatitis B vaccine was administered to 25 nonresponders to the plasma-derived hepatitis B vaccine . After three 10-micrograms doses, nine subjects (36%) produced levels of antibodies to hepatitis B surface antigen (anti-HBs) of less than 2.1 sample ratio units (SRU) (nonresponders), and five (20%) developed anti-HBs of 2.1 to 9.9 SRU (hyporesponders); anti-HBs levels of 10 SRU or greater were detected at least once in 11 vaccinees (44%), but by the sixth and 12th months after the last vaccination, only three and one of these "responders," respectively, still maintained anti-HBs values of 10 SRU or greater . In these 25 subjects HLA subtyping showed a high prevalence of DR7, B8, and the combinations of DR3 and DR7 and DR4 and DR7 . Our findings indicate that the yeast recombinant hepatitis B vaccine was not effective in eliciting a sustained anti-HBs response in nonresponders to the plasma hepatitis B vaccine. Nature, 1988 Sep 15, 335(6187), 251 - 4 Activation at M-phase of a protein kinase encoded by a starfish homologue of the cell cycle control gene cdc2+; Labbe JC et al.; In both starfish and amphibian oocytes, the activity of a major protein kinase which is independent of Ca2+ and cyclic nucleotides increases dramatically at meiotic and mitotic nuclear divisions . The in vivo substrates of this kinase are unknown, but phosphorylation of H1 histone can be used as an in vitro assay . We have purified this kinase from starfish oocytes . The major band in the most highly purified preparation contained a polypeptide of relative molecular mass (Mr) 34,000 (34K) . This is the same size as the protein kinase encoded by cdc2+, which regulates entry into mitosis in fission yeast and is a component of MPF purified from Xenopus . Here, we show that antibodies against p34 recognize the starfish 34K protein and propose that entry into meiotic and mitotic nuclear divisions involves activation of the protein kinase encoded by a homologue of cdc2+ . Given the wide occurrence of cdc2+ homologues from budding yeast to Xenopus and human cells, this activation may act as a common mechanism controlling entry into mitosis in eukaryotic cells. Eur J Biochem, 1988 Sep 15, 176(2), 281 - 5 Diethylstilbestrol . Interactions with membranes and proteins and the different effects upon Ca2+- and Mg2+-dependent activities of the F1-ATPase from Rhodospirillum rubrum; Strid A et al.; The hydrophobic compound diethylstilbestrol inhibits the generation of the proton gradient and the membrane potential in chromatophores from Rhodospirillum rubum and dissipates proton gradients over asolectin vesicle membranes . The Ca2+-ATPase activity of chromatophores, of purified F0F1-ATPase and of purified F1-ATPase is also decreased in the presence of diethylstilbestrol . Other repressed activities are the pyrophosphatase activity of soluble pyrophosphatase from yeast and the NADH oxidation by L-lactate:NAD oxidoreductase . We have previously reported that also ATP synthesis, PPi synthesis and PPi hydrolysis of R . rubrum chromatophores are inhibited by diethylstilbestrol {Strid et al . (1987) Biochim . Biophys . Acta 892, 236-244} . Addition of bovine serum albumin reverses or prevents diethylstilbestrol-induced inhibition of the activities tested . On the other hand, the Mg2+-ATPase activity of chromatophores, purified F0F1-ATPase and purified F1-ATPase are stimulated by low concentrations of diethylstilbestrol . On the basis of its hydrophobicity and the reversal of its inhibition by bovine serum albumin, diethylstilbestrol is proposed to act unspecifically on membranes and at hydrophobic domains of proteins . Such an attack upon the subunits of the F1-ATPase, altering the subunit interactions, is proposed to explain the different results obtained for the Ca2+-ATPase and the Mg2+-ATPase. Biochemistry, 1988 Sep 6, 27(18), 7091 - 6 Glutathione reductase: solvent equilibrium and kinetic isotope effects; Wong KK et al.; Glutathione reductase catalyzes the NADPH-dependent reduction of oxidized glutathione (GSSG) . The kinetic mechanism is ping-pong, and we have investigated the rate-limiting nature of proton-transfer steps in the reactions catalyzed by the spinach, yeast, and human erythrocyte glutathione reductases using a combination of alternate substrate and solvent kinetic isotope effects . With NADPH or GSSG as the variable substrate, at a fixed, saturating concentration of the other substrate, solvent kinetic isotope effects were observed on V but not V/K . Plots of Vm vs mole fraction of D2O (proton inventories) were linear in both cases for the yeast, spinach, and human erythrocyte enzymes . When solvent kinetic isotope effect studies were performed with DTNB instead of GSSG as an alternate substrate, a solvent kinetic isotope effect of 1.0 was observed . Solvent kinetic isotope effect measurements were also performed on the asymmetric disulfides GSSNB and GSSNP by using human erythrocyte glutathione reductase . The Km values for GSSNB and GSSNP were 70 microM and 13 microM, respectively, and V values were 62 and 57% of the one calculated for GSSG, respectively . Both of these substrates yield solvent kinetic isotope effects greater than 1.0 on both V and V/K and linear proton inventories, indicating that a single proton-transfer step is still rate limiting . These data are discussed in relationship to the chemical mechanism of GSSG reduction and the identity of the proton-transfer step whose rate is sensitive to solvent isotopic composition . Finally, the solvent equilibrium isotope effect measured with yeast glutathione reductase is 4.98, which allows us to calculate a fractionation factor for the thiol moiety of GSH of 0.456. Infect Immun, 1988 Sep, 56(9), 2350 - 5 Protective immunity in murine histoplasmosis: functional comparison of adoptively transferred T-cell clones and splenic T cells; Deepe GS Jr; In this study, I examined whether a murine T-cell line and three clones that recognize Histoplasma capsulatum antigens in vitro could confer protection in vivo against a challenge of Histoplasma yeasts . C57BL/6 mice were each inoculated with 5 X 10(4) yeasts intravenously; 1 h later, 5 X 10(6) or 2 X 10(7) resting T cells were inoculated intravenously . At week 1 of infection, the T-cell line and all clones failed to reduce the number of H . capsulatum CFU in the spleens of mice compared with numbers in infected controls . Administration of recombinant interleukin 2 or cyclophosphamide to infected mice did not potentiate the functional activity in vivo of either the T-cell line or the clones . In contrast, inoculation with 2 X 10(7) CD4+ but not CD8+ cells isolated from the spleens of mice immunized with 10(6) viable yeast cells sharply diminished the number of CFU in the spleens of infected animals . Moreover, splenic CD4+ cells from immune mice transferred a delayed-type hypersensitivity response, whereas the T-cell line and clones did not . Injection Injection of an equal number of cloned T cells and CD8+ splenocytes from immune mice did not transfer resistance to infected mice . Additional studies were undertaken to determine if the ineffectiveness of cloned T cells was associated with a failure to migrate to and survive within spleens of infected mice . B6.PL Thy-1a/Cy mice, which are genetically identical to C57BL/6 mice except that T cells of the former bear Thy-1.1 rather than Thy-1.2, were inoculated with Histoplasma yeasts and then injected with immune CD4+ splenocytes or a T-cell clone . At days 1 and 7 of infection, virtually no Thy-1.2+ cells were detected in the spleens of infected mice given cloned T cells . However, the spleens of animals inoculated with immune CD4+ cells contained a small but significant (P less than 0.01) proportion of Thy-1.2+ cells at both day 1 and day 7 postinoculation of H . capsulatum . Thus, the failure of T-cell clones to transfer protection against H . capsulatum may be explained by defective trafficking or poor survival in vivo or both. Anal Biochem, 1988 Sep, 173(2), 405 - 11 Two-step purification and N-terminal amino acid sequence analysis of the rat Mr 90,000 heat shock protein; Denis M; A fast and efficient method for the isolation of the rat Mr approximately 90,000 heat shock protein is presented, comprising a two-step high-performance anion-exchange and gel-permeation column chromatography . The Mr approximately 90,000 protein was purified to electrophoretic homogeneity in high yield (up to 2 mg per 10g of normal rat liver) in 4-5 h . The purified protein was recognized on protein immunoblots by monospecific rabbit antibodies raised against the rat glucocorticoid receptor-associated Mr approximately 90,000 non-ligand-binding protein . The N-terminal sequence of the first 25 amino acids of the purified protein showed a high degree of similarity with Mr approximately 90,000 heat shock proteins from calf, human, Drosophila, and yeast. J Clin Microbiol, 1988 Sep, 26(9), 1861 - 3 Simple method of inducing sporulation by Apophysomyces elegans and Saksenaea vasiformis; Padhye AA et al.; Apophysomyces elegans and Saksenaea vasiformis are notorious for their failure to sporulate on routine media . Agar blocks, permeated with the mycelia of A . elegans and S . vasiformis, were cut aseptically from 7-day-old colonies grown on Sabouraud dextrose agar and transferred to plates containing 20 ml of sterile distilled water supplemented with 0.2 ml of 10% filter-sterilized yeast extract solution . When the plates were incubated at 37 degrees C, all 5 isolates of A . elegans and all 10 isolates of S . vasiformis produced abundant, characteristic sporangia within 7 to 10 days . The method is simple to use and yields consistent results. J Clin Microbiol, 1988 Sep, 26(9), 1814 - 7 Effects of transport temperature and medium on recovery of Bordetella pertussis from nasopharyngeal swabs; Morrill WE et al.; We compared relative recoveries of Bordetella pertussis from simulated nasopharyngeal (NP) specimens incubated in three separate transport media at different temperatures . Transport media included one-half-strength Regan-Lowe (RL.5), Regan-Lowe with one-half-strength agar (RL.5A), and buffered charcoal-yeast extract agar supplemented with alpha-ketoglutarate, lincomycin, and anisomycin (BCYE alpha LA) . For each transport medium, recovery of B . pertussis was least efficient after storage at 25 degrees C . The highest recovery of B . pertussis from a mixed culture was achieved with RL.5 at 4 degrees C . Overall, RL.5 and RL.5A were comparable as transport media whether held at 4 or 25 degrees C, but fewer organisms were recovered from BCYE alpha LA . In addition, Regan-Lowe (RL), Bordet-Gengou, and cyclodextrin media were compared as primary isolation media for recovering B . pertussis from simulated NP swabs held at 4 and 35 degrees C in RL.5 medium . The highest recovery of B . pertussis was obtained on RL primary isolation medium . Bordet-Gengou medium recovered only 80% and cyclodextrin medium recovered less than 60% of the numbers recovered on RL medium . Based on these results, refrigeration (4 degrees C) of NP swabs shipped in RL.5 transport medium and using RL as the primary isolation medium are recommended for recovering B . pertussis from swab specimens. Biol Trace Elem Res, 1988 Sep-Dec, 17, 229 - 36 Chromium induced clinical improvement in symptomatic hypoglycemia; Clausen J; The present study included 20 patients indicating clinical symptoms of hypoglycemia, of which 19 showed a minimal glucose level in the tolerance curve above 2.2 mmol/L (limit for glucose induced hypoglycemia) . This clinical state is defined here as "symptomatic hypoglycemia." All individuals were studied for effects of a daily intake for three mo of a yeast chromium supplement (125 micrograms Cr/d) . The patients were followed by means of their oral glucose tolerance curve (1 g glucose/kg body wt) and by an interrogation scheme prior to during and after chromium supplementation . During three mo of chromium supplementation, a decrease was found in the negative part of the glucose tolerance curve, i.e., the part of the curve being below the fasting level in eight patients (40%) . However, one mo after treatment, 10 patients out of 13 (77%) showed decreased areas of the negative part of the glucose tolerance curve compared to the values during treatment, and 11 out of 15 (73%) showed decreases in the negative part of the curve when post-treatment data were compared to ante-treatment data . The subjective clinical effects were followed by means of a questionnaire . Subjectively, the effects of organo-chromium were especially pronounced on chilliness . Thus seven (47%) indicated improvement and two (15%) indicated that the chilliness disappeared . However, trembling, emotional instability, and disorientation symptoms improved as well. Southeast Asian J Trop Med Public Health, 1988 Sep, 19(3), 501 - 10 Clinical experience with a recombinant DNA hepatitis B vaccine; Andre FE; The clinical testing of EngerixR-B, the hepatitis B vaccine produced by SmithKline Biologicals using recombinant DNA technology, started in February 1984 . Since extensive pre-clinical laboratory work had established that the polypeptide (HBsAg) expressed in genetically engineered yeast cells was after purification--physically, chemically and antigenically similar to the viral surface antigen particles found in the blood of chronic carriers, the aims of the clinical trials were to compare the safety, reactogenicity, immunogenicity and protective efficacy of yeast-derived (YDV) and plasma-derived (PDV) vaccines . By September 1987, 89 studies had been initiated involving a total of 10,545 subjects aged from birth to 82 years . This extensive experience has established that the risk of hypersensitivity to yeast-derived contaminants is negligible since no hypersensitivity reaction has been observed in any vaccinee, the incidence and severity of local reactions have not increased after repeated inoculations and no anti-yeast antibodies were produced by vaccination . Reactogenicity has been comparable to that of PDV's consisting essentially of transient mild irritation at the site of injection presumably caused by the aluminium hydroxide used as adjuvant . The anti-HBs responses to YDV and PDV's were quantitatively (seroconversion rates, peak antibody levels and persistence) as well as qualitatively (epitope specificity and affinity) similar . The expected protective effect of the immune response to the vaccine was confirmed in a challenge study in chimpanzees and in vaccinated human populations (male homosexuals, institutionalized mentally retarded patients, neonates of carrier women) with historically a high infection rate. Anal Biochem, 1988 Aug 15, 173(1), 166 - 73 An in vitro binding assay for angiogenin using placental ribonuclease inhibitor; Bond MD; A convenient in vitro assay for angiogenin has been developed which greatly facilitates its routine detection and quantitation . The assay is based on the capacity of angiogenin to bind placental ribonuclease inhibitor (PRI); it is less tedious and more versatile than existing procedures that measure blood vessel growth or cleavage of rRNA . The test sample is added to a reaction mixture containing a known quantity of PRI, which complexes any angiogenin present in the sample . A slight excess of RNase A, relative to PRI, is then added, and the amount of RNase A which remains unbound is determined by measuring the generation of acid-soluble fragments from yeast RNA . The assay is sensitive to 30 fmol of angiogenin and is linear over a 17-fold concentration range . Use of the binding assay in parallel with a conventional RNase A assay provides a means of detecting angiogenin in chromatographic fractions and differentiating it from RNases . This procedure makes possible the isolation of angiogenin from new sources, such as nonhuman sera . It may also be applicable to other biologically active proteins with sequence homology to RNase A, e.g., eosinophil cationic protein or eosinophil derived neurotoxin. Arch Biochem Biophys, 1988 Aug 15, 265(1), 91 - 3 Phosphofructokinase is responsible for the fructose 2,6-bisphosphate inhibition of hexokinase in tissue extracts; Niemeyer H et al.; Mammalian and yeast hexokinases were reported to be reversibly inhibited by fructose 2,6-bisphosphate in the presence of cytosolic proteins (H . Niemeyer, C . Cerpa, and E . Rabajille (1987) Arch . Biochem . Biophys . 257, 17-26) . Reinvestigation of this finding using a radioassay with {14C}glucose as substrate showed no effect of fructose 2,6-bisphosphate on hexokinase activity of rat liver cytosols . Detailed reexamination of the spectrophotometric assay resulted in the observation that the fructose 2,6-bisphosphate-dependent inhibition was a function of the cytosolic phosphoglucose isomerase and phosphofructokinase activities compared to the amount of glucose-6-phosphate dehydrogenase used as auxiliary enzyme . The diminution or loss of the fructose 2,6-bisphosphate-dependent inhibition produced in aged cytosols was restored by addition of crystalline muscle phosphofructokinase, as well as by decreasing the amount of glucose-6-phosphate dehydrogenase in the assay . When phosphoglucose isomerase, phosphofructokinase, and hexokinase activities were separated by DEAE-chromatography of liver cytosol, no fructose 2,6-bisphosphate-dependent inhibition of hexokinase was found in any single fraction of the chromatogram . However, combination of fractions containing both phosphoglucose isomerase and phosphofructokinase displayed the fructose 2,6-bisphosphate-dependent inhibition on either endogenous hexokinase or added yeast hexokinase . From these results we conclude that the activation of phosphofructokinase elicited by fructose 2,6-bisphosphate is responsible for the hexokinase inhibition observed in the coupled spectrophotometric assay. J Protein Chem, 1988 Aug, 7(4), 491 - 507 Prediction of probable pathways of folding in globular proteins; Kikuchi T et al.; A method is described for the prediction of probable folding pathways of globular proteins, based on the analysis of distance maps . It is applicable to proteins of unknown spatial structure but known amino acid sequence as well as to proteins of known structure . It is based on an objective procedure for the determination of the boundary of compact regions that contain high densities of interresidue contacts on the distance map of a globular protein . The procedure can be used both with contact maps derived from a known three-dimensional protein structure and with predicted contact maps computed by means of a statistical procedure from the amino acid sequence alone . The computed contact map can also be used to predict the location of compact short-range structures, viz . alpha-helices and beta-turns, thereby complementing other statistical predictive procedures . The method provides an objective basis for the derivation of a theoretically predicted pathway of protein folding, proposed by us earlier {Tanaka and Scheraga (1977) Macromolecules 10, 291-304; Nemethy and Scheraga (1979) Proc . Natl . Acad . Sci., U.S.A . 76, 6050-6054}. J Protein Chem, 1988 Aug, 7(4), 427 - 71 Prediction of the location of structural domains in globular proteins; Kikuchi T et al.; The location of structural domains in proteins is predicted from the amino acid sequence, based on the analysis of a computed contact map for the protein, the average distance map (ADM) . Interactions between residues i and j in a protein are subdivided into several ranges, according to the separation {i-j{ in the amino acid sequence . Within each range, average spatial distances between every pair of amino acid residues are computed from a data base of known protein structures . Infrequently occurring pairs are omitted as being statistically insignificant . The average distances are used to construct a predicted ADM . The ADM is analyzed for the occurrence of regions with high densities of contacts (compact regions) . Locations of rapid changes of density between various parts of the map are determined by the use of scanning plots of contact densities . These locations serve to pinpoint the distribution of compact regions . This distribution, in turn, is used to predict boundaries of domains in the protein . The technique provides an objective method for the location of domains both on a contact map derived from a known three-dimensional protein structure, the real distance map (RDM), and on an ADM . While most other published methods for the identification of domains locate them in the known three-dimensional structure of a protein, the technique presented here also permits the prediction of domains in proteins of unknown spatial structure, as the construction of the ADM for a given protein requires knowledge of only its amino acid sequence. Res Commun Chem Pathol Pharmacol, 1988 Aug, 61(2), 167 - 83 Research on heterocyclic compounds . XXIV . Antiinflammatory activity of some imidazo{1,2-c}pyrimidine derivatives; Abignente E et al.; A group of four acidic imidazo{1,2-c}pyrimidine derivatives were subjected to a series of tests in vivo in order to assess their pharmacological activity . Antiinflammatory activity was studied by means of the carrageenin-induced paw edema and pleurisy in rats . Hot plate test and writhing induced by acetic acid in mice were employed to evaluate analgesic activity, whereas the yeast-induced hyperthermia in rats was used to study antipyretic activity . The irritative and ulcerogenic action on the rat gastric mucosa was examined at higher doses . The inhibitory activity on the platelet malondialdehyde production was studied in order to obtain information about the mechanism of action of test compounds. Eur J Cell Biol, 1988 Aug, 46(3), 499 - 505 The changing distribution of actin and nuclear behavior during the cell cycle of the mite-pathogenic fungus Neozygites sp; Butt TM et al.; We have analyzed the behavior of nuclei and actin during the cell cycle of Neozygites sp . with mithramycin and rhodamine-labeled phalloidin . This fungus is an entomophagous zygomycete which grows as a rod-shaped fission yeast containing 2 to 12, mostly 3 to 4, nuclei per cell . The cell cycle is regulated such that there is not a constant nucleus-to-cytoplasmic volume ratio, and mitosis is initiated slightly asynchronously from one end of the cell . During interphase, detected actin occurs exclusively as peripheral plaques, which are most abundant at growing cell tips, and as perinuclear shells . Because the shells disperse and reform concomitantly with the formation and breakdown of a new septum-associated actin array, we infer that they are a novel form of actin storage . Intranuclear mitosis occurs in the absence of detectable spindle actin which suggests that actin is not a universal feature of mitotic systems and may be a cytoplasmic contaminant in open spindles of plant cells . Actin is involved in septum synthesis in previously unreported ways . Prior to morphologically detectable septum initiation, a peripheral equatorial band of longitudinal actin filaments assembles and then shortens to a transverse belt at the future site of septum synthesis . We suggest that this actin array recruits and organizes cell wall synthetic complexes for subsequent septum growth . During detectable septum synthesis, the invaginating plasmalemma bears plaques at a similar concentration to those at growing cellular tips. Ecotoxicol Environ Saf, 1988 Aug, 16(1), 25 - 35 Fungal degradation of polyvinyl acetate; Garcia Trejo A; Certain Aspergillus and Penicillium strains isolated from soil grow well and degrade a commercial sample of polyvinyl acetate (PVA, 4.5 g liter-1) when it is used as the only carbon source . These strains showed an increase in dry weight after 11 days of incubation, along with a depletion of carbohydrates, protein, and deoxyribonucleic acid . This was interpreted as an active turnover of the above metabolites during the degradation . This effect was greatly enhanced by equilibrating the carbon:nitrogen ratio by addition of yeast extract in the original culture . The increase in esterase activity and the loss of viscosity were also considered evidence of the fungal degradation . Isolation of the enzyme was attempted, but unsuccessful. Immunology, 1988 Aug, 64(4), 725 - 31 Immunohistochemical detection of deposits of eosinophil-derived neurotoxin and eosinophil peroxidase in the myocardium of patients with Chagas' disease; Molina HA et al.; An immunohistochemical study of eosinophil distribution in the inflammatory cell infiltrates of four different types of myocardial lesions associated with Chagas' disease--caused by Trypanosoma cruzi--showed larger numbers of these cells in areas presenting tissue necrosis and degeneration, most notably in patients with the most severe myocarditis from a histopathological stand-point . Using antisera specific for human eosinophil-derived neurotoxin or eosinophil peroxidase, we detected deposits of these secretion products on myofibres and in the interstitium of chagasic myocardium displaying necrosis and degeneration but rarely in other types of lesions . These deposits were not detectable in the myocardium of non-chagasic patients who had died from myocardial infarction (acute or in the scarring stage) or myocarditis secondary to bacterial endocarditis . When human eosinophil-derived neurotoxin was incubated with myoblast monolayers there was a significant cell injury, detachment and lysis . These effects were abrogated by yeast RNA, added as a competitive ribonuclease substrate, and inhibited by the ribonuclease inhibitor RNasin, suggesting that the ribonuclease activity of the eosinophil-derived neurotoxin was involved in the effect . These results suggest a link between eosinophil infiltration and necrosis in chagasic myocardial lesions and point to EDN, and perhaps other toxic eosinophil secretion products, as possible mediators of tissue damage. Vaccine, 1988 Aug, 6(4), 299 - 303 Recombinant versus plasma-derived hepatitis B vaccines: issues of safety, immunogenicity and cost-effectiveness; Stephenne J; Any attempt to reduce the incidence of hepatitis B on a worldwide scale requires the availability of large quantities of potent, safe and affordable hepatitis B vaccine . However, ongoing doubts or concerns--justified or not--persist about the comparative safety, immunogenicity and cost-effectiveness of commercially available hepatitis B vaccines, whether derived from plasma or produced via recombinant expression systems . This review compares plasma versus recombinant hepatitis B vaccines in terms of these alleged differences and in the light of increasing clinical data acquired following administration of recombinant yeast-derived hepatitis B vaccines. Cutis . 1988 Aug;42(2):146. Short-term treatment of dandruff with a combination of propylene glycol solution and shampoo; Faergemann J; Many studies now indicate an association between the lipophilic dimorphic yeast Pityrosporon ovale and dandruff . Propylene glycol has been proved effective in the treatment of pityriasis versicolor, Pityrosporon folliculitis, and seborrheic dermatitis . The patients in this study found treatment with a combination of propylene glycol, ethanol, and water, used with a shampoo, to be effective, easy to use, and cosmetically attractive . No side effects were observed by the patients. Proc Natl Acad Sci U S A, 1988 Aug, 85(16), 5849 - 53 Specific dephosphorylation of phosphoproteins by protein-serine and -tyrosine kinases; Kole HK et al.; Five protein kinases are shown to serve as specific phosphatases in the absence of ADP . Although the rates of hydrolysis are very slow compared to the forward phosphorylation rates under optimal conditions, they are of the same order as the reverse reaction in the presence of ADP . Because cells contain approximately equal to 3 mM ATP, neither the reverse reaction nor the phosphatase is likely to play a physiological role . beta-casein B phosphorylated by the catalytic subunit of cAMP-dependent protein kinase (protein kinase A) is specifically dephosphorylated by protein kinase A but not by polypeptide-dependent protein kinase (protein kinase P) . beta-casein B phosphorylated by protein kinase P is specifically dephosphorylated by protein kinase P but not by protein kinase A . Histone H1 phosphorylated by protein kinase C is dephosphorylated by the same enzyme in the absence of ADP . In all cases tested addition of ADP and F1-ATPase accelerates moderately the rate of dephosphorylation . Native H+-ATPase from yeast plasma membranes is isolated mainly in the phosphorylated form . It is dephosphorylated and rephosphorylated by protein kinase P but not by protein kinase A . Protein-tyrosine kinase of the epidermal growth factor receptor phosphorylates the random synthetic polypeptide poly(Glu80Tyr20) . The phosphorylated polymer is specifically dephosphorylated in the absence of ADP by epidermal growth factor receptor preparations but not by insulin receptor preparations . The same polymer phosphorylated by insulin receptor is dephosphorylated by insulin receptor but not by epidermal growth factor receptor preparations . By using a cycle of dephosphorylation-rephosphorylation, it is possible to identify proteins that are phosphorylated by these protein kinases in vivo . Should this method be applicable to additional protein kinases, it should be possible to estimate the quantitative contribution of each protein kinase to a single phosphoprotein. Int J Radiat Biol, 1988 Aug, 54(2), 285 - 98 DNA damage does not appear to be a trigger for thermotolerance in mammalian cells; Anderson RL et al.; The hypothesis that DNA damage is the trigger for thermotolerance in mammalian cells was tested in Chinese hamster ovary cells by looking for evidence of thermotolerance after ionizing radiation or ultraviolet light exposure . As previous studies have demonstrated that relatively non-toxic radiation exposures do not induce thermotolerance in mammalian cells (Li et al . 1976), higher doses, comparable to those used in yeast to induce thermotolerance (Mitchel and Morrison 1984), were tested in this study . Doses of this magnitude are lethal to mammalian cells, thereby precluding the use of clonogenic survival as an endpoint . We therefore used three alternative assays which are indicators of the subsequent development of thermotolerance . These were; (a) heat-induced inhibition of total protein synthesis, (b) heat-induced uptake of dansyl lysine, and (c) synthesis of heat shock proteins . Only total protein synthesis revealed evidence of a small degree of thermotolerance which occurred immediately after ionizing radiation exposure . By 4 h postirradiation the tolerance, as measured by this assay, was no longer evident . No evidence of thermotolerance was seen following UV exposure . In addition, when a large radiation dose was given either immediately before or after a heat treatment used to induce thermotolerance, there was no alteration in the level of heat-induced tolerance, despite the extensive number of DNA stand breaks caused by the radiation . Our data therefore suggest that, in mammalian cells, the type of DNA damage caused by ionizing radiation is not the trigger for the induction of thermotolerance. Vaccine, 1988 Aug, 6(4), 304 - 6 Polyvalent recombinant antigens: a new vaccine strategy; Kingsman SM et al.; Recombinant DNA technology can be used to produce individual proteins from infectious organisms and these have the potential to be used as subunit vaccines . In many cases however these proteins display a low immunogenicity . One reason for this might be the poor presentation of the antigen to the cells of the immune system . A number of new recombinant DNA approaches are now being tried to improve the production and the presentation of antigens . One method is to use a carrier protein encoded by a retrotransposon of yeast called the Ty element . This protein can be linked to a wide range of viral antigens via the construction of hybrid genes . The fusion proteins assemble in yeast into 60 nm virus-like particles (VLPs) that display the additional antigens on the surface . These hybrid Ty-VLPs are good immunogens even in the absence of adjuvant . Similar approaches to the production of polyvalent particulate antigens involve using carrier proteins derived from hepatitis B and polio viruses. Cell, 1988 Jul 29, 54(3), 423 - 31 The Xenopus cdc2 protein is a component of MPF, a cytoplasmic regulator of mitosis; Dunphy WG et al.; In Xenopus, a cytoplasmic agent known as MPF induces entry into mitosis . In fission yeast, genetic studies have shown that the cdc2 kinase regulates mitotic initiation . The 13 kd product of the suc1 gene interacts with the cdc2 kinase in yeast cells . We show that the yeast suc1 gene product (p13) is a potent inhibitor of MPF in cell-free extracts from Xenopus eggs . p13 appears to exert its antagonistic effect by binding directly to MPF . MPF activity is quantitatively depleted by chromatography on a p13 affinity column . Concomitantly, the Xenopus counterpart of the yeast cdc2 protein is adsorbed to the column . A 42 kd protein also binds specifically to the p13 affinity matrix . These findings suggest that the Xenopus cdc2 protein and the 42 kd protein are components of MPF. Cell, 1988 Jul 15, 54(2), 199 - 207 The hormone-binding domains of the estrogen and glucocorticoid receptors contain an inducible transcription activation function; Webster NJ et al.; We have constructed novel chimeric receptors, GAL-ER and GAL-GR, consisting of the DNA-binding domain of the yeast transcription factor GAL4 joined to the C-terminal region containing the hormone-binding domain of either the human estrogen (hER) or human glucocorticoid (hGR) receptor . Stimulation of transcription by GAL-ER and GAL-GR from GAL4-responsive reporter genes was hormone dependent, and the activation function has been localized to the hER or hGR region . Both chimeric receptors recognized GAL4-responsive elements only in the presence of hormone or anti-hormone, yet solely the hormone was capable of stimulating transcription . These and additional results suggest that the hormone plays at least a dual role . First, the hormone, or anti-hormone, is responsible for receptor "transformation" allowing the recognition of responsive DNA elements . Second, the hormone, but not the anti-hormone, can induce a transcription activation function present in the region that contains the hormone-binding domain. Gene, 1988 Jul 15, 67(1), 1 - 11 Isolation of genes expressed in specific tissues of Arabidopsis thaliana by differential screening of a genomic library; Simoens CR et al.; The small genome size of Arabidopsis thaliana allows the isolation of genes expressed in specific tissues and under controlled conditions by the differential screening of a genomic library, as has been shown previously for yeast and Drosophila . cDNA probes, based on poly(A)+ mRNA isolated from different Arabidopsis organs, were used in colony hybridizations with 1145 randomly chosen genomic clones, representing 27,000 kb of Arabidopsis DNA . Twenty percent of the clones containing low-copy-number sequences hybridized with one or more of the cDNA probes that were synthesized from mRNA isolated from leaves, stems, seed pods, inflorescences, callus tissue, and light-grown and dark-grown plants . Comparison of the colony hybridizations led to the identification of a large variety of clones which contain differentially expressed genes . The pattern of expression was confirmed by Northern analysis . The advantage of the described method is that it yields directly genomic sequences that contain specifically expressed or induced genes . In particular, it circumvents the construction and differential screening of cDNA libraries for every tissue or environmental parameter to be analyzed. Biochim Biophys Acta, 1988 Jul 13, 950(2), 118 - 31 Interactions of porphyrins and transfer RNA; Foster N et al.; The interactions of the free base porphyrin, tetra-(4N-methylpyridyl)porphyrin and its copper(II), manganese(III) and zinc(II) complexes with brewer's yeast type V phenylalaninyl tRNA were evaluated by UV-visible spectroscopy, circular dichroism and melting temperature studies over a range of magnesium ion concentrations and ionic strengths . Scatchard analysis of absorption spectra of the porphyrins in the presence of tRNA showed the free base, copper and zinc porphyrins to have binding constants of 7.3 X 10(7), 1.7 X 10(6) and 2.3 X 10(8), respectively; the manganese(III) complex did not demonstrate changes in its electronic spectra that enable the calculation of a binding constant . The results of the spectroscopic studies indicate a mode of binding for the free base, copper(II) and zinc(II) complexes that is neither intercalative nor simply outside electrostatic . The magnitude of the binding constants and the UV-visible results support intercalation, but the analyses of the thermal denaturation studies and the circular dichroism evaluations suggest that the porphyrins are associating at a single site in a fold of the tertiary structure of the tRNA close to several crucial hydrogen bonds, perhaps in the vicinity of the P10 loop . That the manganese(III) complex does not bind in this site points to constraints on the axial thickness of a molecule that may be accommodated in this locus. Nucleic Acids Res, 1988 Jul 11, 16(13), 6015 - 25 Pulsed-field electrophoresis indicates larger-than-expected sizes for mycoplasma genomes; Pyle LE et al.; The sizes of large DNA fragments produced from genomes of members of the Mycoplasmataceae by digestion with restriction endonucleases having infrequent (1 to 3) cleavage sites within the genome were estimated from their mobility in contour-clamped homogeneous electric field (CHEF) agarose gel electrophoresis by comparison with yeast chromosomal DNA markers . The estimates of total genome size for 7 strains of 6 species ranged from approximately 900 kilo base pairs (kb) for Ureaplasma urealyticum 960T to 1330 kb for M . mycoides subsp . mycoides, GC-1176 . The values derived from this new method are considerably higher than those of approximately 500 Mdaltons or 750 kb previously reported for genome sizes in members of the Mycoplasmataceae. J Mol Biol, 1988 Jul 5, 202(1), 161 - 8 Structure and expression of an actin gene of Physarum polycephalum; Gonzalez-y-Merchand JA et al.; Physarum polycephalum (strain M3CVIII) contains four unlinked actin gene loci, each with two alleles (ardA1, ardA2, ardB1, ardB2, ardC1, ardC2, ardD1 and ardD2) . The 4800 base HindIII fragment of the ardC2 allele was previously isolated as a recombinant phage lambda . We now report the structure of the actin gene sequences (C-actin gene) . The gene, which contains four intervening sequences, codes for the principal actin isotype of plasmodia and it is expressed in both the haploid myxamoebal and diploid plasmodial phases of the life cycle . The C-actin isotype is closely related to actins of Dictyostelium, Acanthamoebae, Drosophila, sea urchin and mammalian cytoplasmic actin, and more distantly related to actins of yeast, Entamoebae and Tetrahymena . The ardC1 and ardC2 alleles differ by a 700(+/- 100) base-pair insertion/deletion in the vicinity of the 3' end of the transcribed region of the gene. Proc Natl Acad Sci U S A, 1988 Jul, 85(14), 4971 - 5 Comparison of time-resolved and -unresolved measurements of deoxyhemoglobin in brain; Chance B et al.; Continuous (CW) and pulsed light were used for the noninvasive measurement of hemoglobin oxygenation in tissues . A dual wavelength method of continuous illumination spectroscopy used 760 nm (deoxyhemoglobin peak) and 800 nm (an oxyhemoglobin-deoxyhemoglobin isosbestic point) to measure the kinetics and extent of oxyhemoglobin deoxygenation in brains during mild ischemia/hypoxia . Absorption and scattering were modeled in an artificial milk/yeast blood system, which gave an exponential relationship between absorption and optical path length to a depth of 7 cm . Time-resolved spectroscopy (10-ps resolution) afforded a display of the times and distances of arrival of photons emitted by the cat brain in response to a 10-ps input pulse . The emitted photons rose to a peak in a fraction of a nanosecond and declined exponentially over a few nanoseconds . The half-time of exponential decay corresponds to photon migration over a distance of 4 cm . Exponential light emission continued for several more nanoseconds when the brain was encased by the skull, which plays a key role in prolonging light emission . The exponential decline of light intensity has a value {exp(-microL)}, where L is the path length determined from the time/distance scale and mu is the characteristic of the migration of light in the brain . The factor mu is increased by increasing absorption, and mu' = epsilon C where epsilon and C are the Beer-Lambert parameters of extinction coefficient (epsilon) and concentration (C) . Thus, deoxyhemoglobin can be quantified in brain tissues. J Nutr, 1988 Jul, 118(7), 853 - 8 Effect of selenium repletion on glutathione peroxidase protein level in rat liver; Knight SA et al.; We have previously reported that liver glutathione peroxidase (GSH-Px, EC 1.11.1.9) protein level and activity decrease exponentially during Se deficiency . To determine the effect of Se repletion on these parameters, Se-deficient rats were repleted with 0.1 or 0.5 mg Se/kg diet as Na2SeO3 in a 30% torula yeast-based diet and were killed 0, 1, 2, 3, 5, 7 or 14 d later . GSH-Px protein was quantitated using anti-GSH-Px antibodies . Dietary repletion with 0.5 mg Se/kg diet increased GSH-Px protein and activity significantly (P less than 0.05) after 1 d . After 5 d for GSH-Px protein and 7 d for activity the rate of increase slowed, and at d 14 neither GSH-Px protein nor activity was significantly different from that of Se-adequate rats . Repletion with 0.1 mg Se/kg diet did not significantly increase GSH-Px protein or activity until 14 d . To examine the short-term effect of Se repletion, Se-deficient rats were injected intravenously with 15 or 60 micrograms Se as Na2SeO3 and killed 1, 3, 6, 12 or 24 h later . Only rats injected with 60 micrograms Se and killed 24 h later had a significant increase in GSH-Px activity along with a marginally significant increase in GSH-Px protein . These response curves indicate that homeostatic processes control the level of GSH-Px . The lack of an increase in GSH-Px until 24 h after Se administration implies that additional metabolic events after a rise in cellular Se may be necessary prior to an increase in GSH-Px synthesis in Se-deficient rats. Eur J Biochem, 1988 Jul 1, 174(4), 691 - 7 Uptake and intracellular transport in rat liver of formaldehyde-treated bovine serum albumin labelled with 125I-tyramine-cellobiose; Misquith S et al.; 1 . Endocytosis of formaldehyde-treated bovine serum albumin by rat liver sinusoidal cells has been followed by injecting rats with the protein labelled with 125I-tyramine cellobiose (125I-TCfBSA) . 125I-TCfBSA is quickly taken up by the liver; the radioactivity present in the organ reaches a plateau 5-10 min after injection and is maintained for up to at least 180 min . During the first 5 min most of radioactivity remains acid-precipitable . After which, labelled acid-soluble components are produced at a constant rate for up to 30-40 min . 2 . Differential centrifugation shows that radioactivity is first recovered mainly in the microsomal fraction . Within a few minutes it exhibits a distribution pattern similar to that of lysosomal enzymes, being chiefly located in the mitochondrial fractions . 3 . Isopycnic centrifugation in a sucrose gradient of the microsomal fraction isolated 1 min after injection indicates a similar distribution for radioactivity and alkaline phosphodiesterase . Later, the microsomal radioactivity distribution curve is shifted towards higher densities and becomes distinct from that of the plasma-membrane enzyme . After isopycnic centrifugation in a sucrose gradient of the total mitochondrial fraction a considerable overlapping of acid-precipitable and acid-soluble radioactivity distributions is observed without significant changes with time . The same is observed in a Percoll gradient except that after a relatively long time (greater than 30 min) of injection a marked shift of radioactivity distribution towards higher densities occurs . 4 . A pretreatment of rats with Triton WR 1339, a density perturbant of liver lysosomes, causes a striking shift of acid-soluble radioactivity distribution in a sucrose gradient towards lower densities while having markedly less influence on the acid-precipitable distribution . As a result, a distinction between the distribution of both kinds of radioactivity becomes clearly apparent . A preinjection of yeast invertase, modifies the acid-soluble distribution without having a significant effect on the acid-precipitable distribution up to 30 min after 125I-TCfBSA injection . 5 . Glycyl-1-phenylalanine-2-naphthylamide largely releases acid-soluble radioactivity associated with the mitochondrial fraction, whatever the time after 125I-TCfBSA injection . On the other hand the proportion of acid-precipitable radioactivity present in the fraction that can be released is almost zero at 10 min after injection, and it later increases . 6 . The results presented here are best explained by supposing that, after being trapped in small pinocytic vesicles, 125I-TCfBSA is quickly delivered to the endosomes.(ABSTRACT TRUNCATED AT 400 WORDS) Proc Natl Acad Sci U S A, 1988 Jul, 85(13), 4643 - 7 Posttranslational modification of the Ha-ras oncogene protein: evidence for a third class of protein carboxyl methyltransferases; Clarke S et al.; The ras oncogene products require membrane localization for their function, and this is thought to be accomplished by the addition of a palmitoyl group to a cysteine residue near the carboxyl terminus of the nascent chain . A lipidated carboxyl-terminal cysteine residue is also found in sequence-related yeast sex factors, and in at least two cases, the alpha-carboxyl group is also methyl esterified . To determine if ras proteins are themselves modified by a similar type of methylation reaction, we incubated rat embryo fibroblasts transformed with p53 and activated Ha-ras oncogenes with L-{methyl-3H}methionine under conditions in which the isotope was converted to the methyl donor S-adenosyl-L-{methyl-3H}methionine . By using an assay that detects methyl ester linkages, we found that immunoprecipitated ras proteins are in fact esterified and that the stability of these esters is consistent with a carboxyl-terminal localization . This methylation reaction may be important in regulating the interaction of ras proteins with plasma membrane components . The presence of analogous carboxyl-terminal tetrapeptide sequences in other proteins may provide a general recognition sequence for lipidation and methylation modification reactions. Cell, 1988 Jul 1, 54(1), 17 - 26 Activation of cdc2 protein kinase during mitosis in human cells: cell cycle-dependent phosphorylation and subunit rearrangement; Draetta G et al.; HeLa cell p34, homolog of the yeast cdc2+/CDC28 protein kinase, has been investigated . p34 was phosphorylated at two or more sites and existed in a complex with p13, the previously identified homolog of the suc1+ gene product of S . pombe . A fraction of the most highly phosphorylated form of p34 was also associated with p62, a newly identified protein that became phosphorylated in vitro . The phosphorylation state of p34, its association with p62, and the protein kinase activity of the complex were each subject to cell cycle regulation . In newly born cells early in G1, p34 was unphosphorylated, not associated with p62, and inactive as a protein kinase . Each of these conditions was reversed in G2 and the p34/p62 complex was maximally active as a protein kinase, with respect to both endogenous and exogenous substrates, during mitotic metaphase . p34 may act to regulate the G2/M transition in HeLa cells. J Ethnopharmacol, 1988 Jul-Aug, 23(2-3), 285 - 90 Antipyretic activity of alpha- and beta-santonin; Martin ML et al.; The antipyretic activity of two sesquiterpene lactones, beta-santonin and arsubin, isolated from the lipid fraction of Artemisia coerulescens subsp . gallica was determined in rats along with the activity of alpha-santonin . In its alpha- and beta-forms, santonin caused a decrease in the body temperature of rats made febrile by the subcutaneous injection of beer yeast . This decrease, more pronounced in the case of beta-santonin, was dose-dependent and antagonized by pretreatment with haloperidol, an agent that also opposes the antipyretic activity of dopamine . These findings seem to show that the alpha- and beta-forms of santonin act on rectal temperature in a way similar to dopamine. J Pharmacobiodyn, 1988 Jul, 11(7), 472 - 8 Local induction of a tumor necrosis factor (TNF)-like cytotoxic factor in murine tissues with tumorous and nontumorous inflammation after systemic administration of antitumor polysaccharides; Takahashi K et al.; Local induction of a cytotoxic factor (CF), which was reported by us to be a tumor necrosis factor (TNF)-like molecule, in murine tumor tissues by i.v . administration of antitumor polysaccharides was studied . The CF was measured by cytolysis assay against L929 fibroblasts in vitro . The antitumor polysaccharides mannoglucan polyalcohol (MGA), lentinan, carboxymethyl-(1----3)-beta-D-linear glucan DP540 (CM-TAK) and yeast mannan induced the CF in MH134 hepatoma tissues inoculated intradermally, with MGA inducing the highest level of the CF . MGA induced the CF in MM46 mammary carcinoma, Ehrlich carcinoma, and MH134 hepatoma, the growth of which were all inhibited by MGA, but not in Lewis lung carcinoma and EL-4 lymphoma, which are therapeutically resistant to MGA . MGA induced the CF in solid MH134 hepatoma tissues inoculated subcutaneously or intramuscularly as well as intradermally, but not in ascitic fluids with intraperitoneal MH134 hepatoma on which MGA is ineffective . These findings suggest that CF induction is correlated with the antitumor activity of polysaccharides . CF induction in tumor tissues was detectable 6 h after i.d . inoculation of MH134 hepatoma . Even in nontumorous inflammatory skin tissues produced by injection of TAK, the CF was induced by MGA . Thus, the early inflammatory reaction with accumulation of host cells and MGA treatment act cooperatively in local induction of the CF. Ann Inst Pasteur Microbiol, 1988 Jul-Aug, 139(4), 393 - 402 Legionella moravica sp . nov . and Legionella brunensis sp . nov . isolated from cooling-tower water; Wilkinson HW et al.; Two Legionella-like organisms were isolated from cooling-tower water samples in Czechoslovakia . They were presumptively identified as legionellae by their growth on buffered charcoal-yeast extract agar (BCYE) containing L-cysteine and their absence of growth on BCYE without L-cysteine . Both strains contained predominately branch-chained cellular fatty acids and were therefore definitively placed in the genus Legionella . They were serologically distinct from other described Legionella species and were shown by DNA studies to constitute two new Legionella species, Legionella moravica (type strain 316-36; ATCC 43877) and Legionella brunensis (type strain 441-1; ATCC 43878). Mycopathologia, 1988 Jul, 103(1), 21 - 7 Paraquat induced thiol modulation of Histoplasma capsulatum morphogenesis; Leith KM et al.; Cysteine metabolism with the subsequent release of anionic thiols has been shown to be involved in yeast cell morphogenesis of the dimorphic, pathogenic fungus Histoplasma capsulatum . Following transfer to fresh medium, intracellular thiol levels during the initial 2-4 h appear to determine the eventual growth form . Mild oxidative stress induced by paraquat (methyl viologen) caused enhanced intracellular and extracellular thiol production and an increase in protein thiol formation . Mildly stressed cells continued to grow in the yeast form . Severe oxidative stress induced by high concentrations of paraquat resulted in lowered thiol production as well as reversion to the alternate mycelial morphology . These results suggest that thiol modulation of intracellular protein status may be involved in morphogenesis of H . capsulatum. J Pharm Sci, 1988 Jul, 77(7), 569 - 70 Kinetics of drug action in disease states . XXVI: Effect of fever on the pharmacodynamics of theophylline-induced seizures in rats; Yasuhara M et al.; This investigation was designed to determine the effect of fever on the neurotoxicity of theophylline as reflected by the concentrations of this drug that cause convulsions in experimental animals . Fever was produced in male, inbred, adult Lewis rats (approximately 180 g) by sc injection of brewer's yeast; an elevation of body temperature of 1.2 +/- 0.4 degrees C (mean +/- SD) was achieved at the time of the pharmacodynamic measurements . Theophylline was infused iv at a rate of 1.03 mg/min until the onset of maximal seizures . Drug concentrations in serum, serum water, brain, and cerebrospinal fluid (CSF) at that time were determined by high-performance liquid chromatography . Compared with the control group, the group of febrile rats had statically significantly lower serum protein concentrations, decreased serum protein binding of theophylline, and slightly increased theophylline concentrations in the CSF at the onset of seizures . Inasmuch as theophylline concentrations in the CSF reflect the concentrations of this drug in the biophase, the results of this study show that fever does not increase the sensitivity of the central nervous system to the neurotoxic effects of theophylline in rats . In fact, a statistically significant positive correlation between theophylline concentrations in the CSF and body temperature was found in this investigation, suggesting a decreased sensitivity of the animals to the neurotoxic effects of theophylline at higher body temperature. J Photochem Photobiol B, 1988 Jul, 2(1), 109 - 22 Photophysical, photochemical and photobiological properties of pyrrolocoumarins; a new class of photoactive compounds; Gibbs NK et al.; With the aim of finding new photoactive compounds that may reduce the side effects of 8-methoxypsoralen photochemotherapy we report on some photophysical, photochemical and photobiological properties of recently synthesized pyrrolocoumarins, in particular 4-methyl-N-ethyl-pyrrolo{3,2-g}coumarin (PCNEt) which has an absorption maximum in the UV-A (320-400 nm) . Laser (347 nm) flash photolysis studies showed triplet transients that were quenched by O2 and by ground state PCNEt . Singlet minus triplet spectra were broad (350-550 nm) and, at 700 nm, indicated solvated electron and radical production . PCNEt complexes with DNA in the dark and photobinds to thymine but does not form DNA cross-links . PCNEt was phototoxic in yeast with an action spectrum similar to its absorption spectrum . PCNEt showed photohaemolytic activity but was not phototoxic on guinea pig skin . These data suggest that PCNEt may photosensitize via several mechanisms: direct DNA photobinding, photodynamic action, or photoproduction of radicals. Cancer Lett, 1988 Jul, 41(1), 21 - 30 Diethylglyoxal bis(guanylhydrazone): a novel highly potent inhibitor of S-adenosylmethionine decarboxylase with promising properties for potential chemotherapeutic use; Elo H et al.; Diethylglyoxal bis(guanylhydrazone) (DEGBG), a novel analog of the antileukemic agent methylglyoxal bis(guanylhydrazone) (MGBG) was synthesized . It was found to be the most powerful inhibitor of yeast S-adenosylmethionine decarboxylase (AdoMetDC) so far studied (Ki approx . 9 nM) . This property, together with the finding that the compound is a weaker inhibitor of intestinal diamine oxidase than are MGBG and its glyoxal, ethylglyoxal and ethylmethylglyoxal analogs, makes the compound a promising candidate as a polyamine antimetabolite for chemotherapy studies . DEGBG was also found to potentiate the antiproliferative effect of the ornithine decarboxylase inhibitor alpha-difluoromethyl ornithine against mouse L1210 leukemia cells in vitro . DEGBG increased several-fold the intracellular putrescine concentration of cultured L1210 cells, just as MGBG and its ethylglyoxal analog are known to do . The results strongly suggest that DEGBG is worth further studies . Combined with previous studies, they also made possible the construction of some empirical rules concerning the structure-activity relationships of bis(guanylhydrazone) type inhibitors of AdoMetDC . The identity of DEGBG was confirmed by a single-crystal X-ray analysis and by 1H- and 13C-NMR spectroscopy . It consisted of the same isomer as MGBG and several of its analogs are known to consist of. J Infect, 1988 Jul, 17(1), 49 - 55 Comparison of results of recombinant and plasma-derived hepatitis B vaccines in Japanese nursery-school children; Hayashi J et al.; Results obtained with a recombinant hepatitis B vaccine were compared with those obtained with a plasma-derived hepatitis B vaccine in separate studies conducted in nursery schools in which at least one child had hepatitis B e antigen associated with surface antigen . Recombinant vaccine (5 micrograms), made in Japan and prepared from antigen expressed in yeast, was given to 118 children (aged 0-5 years of age, mean age 2.9 years) . Plasma-derived vaccine (10 micrograms) was given to 243 children . Side-reactions were not observed with either vaccine . Seroconversion rates for the recombinant vaccine group were 8.5% after 1 month, 98.3% after 9 months and 100% after 12 months . For the plasma-derived vaccine group, the rate after 1 month was 26.3% after 9 months 82.3%, and after 12 months 77.9% . Although in the recombinant vaccine group the immune response developed more slowly during the early phase, seroconversion rates were significantly higher than in the plasma-derived vaccine group after 6 months . Titres of antibodies also were significantly higher in the recombinant vaccine group after the third injection . None of the children in either group became infected with hepatitis B virus . These results confirm the high immunogenicity, safety and efficacy of the recombinant vaccine given to these nursery school children. Postgrad Med, 1988 Jul, 84(1), 85 - 94 Immunoprophylaxis of viral hepatitis; Bruckstein AH; Because safe, effective treatment for established viral hepatitis is not available, physicians need to be acquainted with recent advances in prophylaxis . Immune globulin (Gamastan, Gammar) is used for both preexposure and postexposure prophylaxis of hepatitis A, and side effects are rare . Two vaccines (Heptavax-B, Recombivax HB) are licensed for hepatitis B, one is a plasma-derived vaccine, the other a yeast-recombinant vaccine . Indirectly, these also control hepatitis D (delta agent), which needs hepatitis B virus to develop. Science, 1988 Jun 24, 240(4860), 1759 - 64 The leucine zipper: a hypothetical structure common to a new class of DNA binding proteins; Landschulz WH et al.; A 30-amino-acid segment of C/EBP, a newly discovered enhancer binding protein, shares notable sequence similarity with a segment of the cellular Myc transforming protein . Display of these respective amino acid sequences on an idealized alpha helix revealed a periodic repetition of leucine residues at every seventh position over a distance covering eight helical turns . The periodic array of at least four leucines was also noted in the sequences of the Fos and Jun transforming proteins, as well as that of the yeast gene regulatory protein, GCN4 . The polypeptide segments containing these periodic arrays of leucine residues are proposed to exist in an alpha-helical conformation, and the leucine side chains extending from one alpha helix interdigitate with those displayed from a similar alpha helix of a second polypeptide, facilitating dimerization . This hypothetical structure is referred to as the "leucine zipper," and it may represent a characteristic property of a new category of DNA binding proteins. Biochem Biophys Res Commun, 1988 Jun 16, 153(2), 855 - 61 Effect of selenium status on mRNA levels for glutathione peroxidase in rat liver; Saedi MS et al.; To determine the effect of Se status on the level of mRNA for Se-dependent glutathione peroxidase (EC 1.11.1.9), rats were fed either a Se-deficient torula yeast diet (less than 0.02 mg Se/kg diet) or a Se-adequate diet (+0.2 mg Se/kg as Na2SeO3) for greater than 135 d . Liver glutathione peroxidase activity was 0.025 for Se-deficient versus 0.615 EU/mg protein for Se-adequate rats . Total liver RNA and polyadenylated RNA were isolated and subjected to Northern blot analysis using a 700 bp DNA probe from cloned murine glutathione peroxidase . Autoradiography showed that Se-deficient liver had 7-17% of the mRNA for glutathione peroxidase present in Se-adequate liver, suggesting that Se status may regulate the level of mRNA for this selenoenzyme. Nature, 1988 Jun 16, 333(6174), 683 - 6 Crystal structure of enolase indicates that enolase and pyruvate kinase evolved from a common ancestor; Lebioda L et al.; Enolase or 2-phospho-D-glycerate hydrolase catalyses the dehydration of 2-phosphoglycerate to phosphoenolpyruvate, which in turn is converted by pyruvate kinase to pyruvate . We describe here the crystallographic determination of the structure of yeast enolase at high resolution (2.25 A) and an analysis of the structural homology between enolase, pyruvate kinase and triose phosphate isomerase . Each of the two subunits of enolase forms two distinctive domains . The larger domain (residues 143-420) is a regular 8-fold beta/alpha-barrel, as first found in triose phosphate isomerase, and later in pyruvate kinase and 11 other functionally different enzymes . An analysis of the molecular geometries of enolase and pyruvate kinase based on the roughly 8-fold symmetry of the barrel showed a structural homology better than expected for proteins related by convergent evolution . We argue that enolase and pyruvate kinase have evolved from a common ancestral multifunctional enzyme which could process phosphoenolpyruvate in both directions along the glycolytic pathway . There is structural and sequence evidence that muconate lactonizing enzyme later evolved from enolase. Nature, 1988 Jun 16, 333(6174), 676 - 9 Regulated expression and phosphorylation of a possible mammalian cell-cycle control protein; Lee MG et al.; A novel approach to the study of the control of the mammalian cell cycle was opened by the cloning of a human gene by complementation of a fission-yeast cdc2 cell-cycle mutant . We have investigated the behaviour of the RNA and protein products of this human gene, CDC2Hs, and its murine equivalent, CDC2Mm during serum starvation and re-feeding of cultured fibroblasts . In contrast to the pattern of wild-type cdc2+ expression in fission yeast previously described, the mammalian homologue displays variation in both RNA and protein levels during exit from and re-entry into the mitotic cycle . Like its yeast counterpart, however, the mammalian CDC2 protein (p34CDC2) becomes dephosphorylated upon shifting from exponential growth to quiescence, and rephosphorylated late in the G1 phase when cells are stimulated to re-enter the cycle . We propose that phosphorylation of p34CDC2 serves as a regulatory mechanism generally in eukaryotic cells, while transcriptional control of the CDC2 gene in higher eukaryotes may be relevant to long term processes such as senescence and differentiation. Biochem J, 1988 Jun 15, 252(3), 791 - 4 Comparison of p25 presequence peptide and melittin . Red blood cell haemolysis and band 3 aggregation; Clague MJ et al.; The 25 residue presequence (p25) for subunit IV of yeast cytochrome oxidase had previously been shown to possess structural and behavioural characteristics in common with the bee venom polypeptide, melittin . The present study extends the results of leakage experiments on model-membrane systems to the haemolysis of human erythrocytes, which both peptides are shown to accomplish in a manner sensitive to membrane potential . In addition, the laser flash-induced transient dichroism technique for measuring protein rotational diffusion has been used to show that both peptides aggregate band 3, the major integral membrane protein of the erythrocyte . Aggregation cannot be reversed by high ionic strength; this serves to differentiate these peptides from other positively charged species such as polylysine that aggregate band 3 at low ionic strength . These results suggest that aggregation of membrane proteins may possibly prove to be a feature of the interaction of p25 signal peptide with mitochondrial membranes. Nucleic Acids Res, 1988 Jun 10, 16(11), 5175 - 90 Inhibition of transcription in eukaryotic cells by X-irradiation: relation to the loss of topological constraint in closed DNA loops; Luchnik AN et al.; X irradiation was found to inhibit in vivo transcription in mammalian, yeast, insect and avian cells in a dose-dependent manner . Measurements of DNA nicking indicated that about one DNA single-strand break per estimated DNA loop (domain) length is sufficient to explain the effect . The inhibitory effect was partially reversed by post-irradiation incubation of cells . During such incubation DNA nicking was considerably repaired . The size of transcripts was not changed by irradiation . The in vitro (run on) activity of RNA polymerase in nuclei isolated from irradiated cells also was not altered . The dose-response curves were different in various cells, correlating with the reported unequal average domain size of supercoiled DNA (and also replicon size) in diverse organisms. Nucleic Acids Res, 1988 Jun 10, 16(11), 4853 - 63 A computer program allows the separation of a wide range of chromosome sizes by pulsed field gel electrophoresis; Sor F; The introduction of Pulsed Field Gel Electrophoresis techniques, which allow the separation of DNA molecules of molecular weights as high as chromosomes of lower eukaryotes, has given a powerful tool to geneticists . The resolution expected from these techniques is dependent on numerous parameters, among them pulse time and field strength . A given set of these parameters allows only a limited range of molecular weights to be resolved . To allow the separation of a broader molecular weight range on a single gel, we designed a computer program, driving a simple switching device, to take care of switching electrodes and power supplies in OFAGE migrations . This program has been designed to be used with any technique calling for periodic switching or inversion of the electric field, and/or variation of the electric field applied during electrophoresis . As an example, we show the results obtained with yeast genera in which chromosome sizes range from 260 to 9,000 kilobase pairs. Presse Med, 1988 Jun 4, 17(22), 1150 - 1 {Immunogenicity of a hepatitis B vaccine obtained by genetic recombination and containing products of S and pre-S2 genes}; Coursaget P et al.; Immunization against hepatitis B by means of a vaccine obtained by genetic recombination on chinese hamster ovary cells was attempted in 32 adult subjects . The HBs antigen was purified from the culture supernatant and contained S and pre S2 genes products . Three 20 micrograms doses were injected intramuscularly at intervals of one month . Anti-HBs seroconversion levels and the geometrical mean of antibodies were slightly higher than those observed with plasma vaccines or genetically engineered yeast-derived vaccine . Antibodies directed against HBs appeared more rapidly and at a higher titre . Anti-pre S2 antibodies were detected after the third injection in 84 per cent of the subjects vaccinated . This recombinant hepatitis B vaccine prepared from chinese hamster ovary cells will probably be as effective as the first generation vaccines obtained by purifying the HBs antigen obtained from the blood of asymptomatic carriers. Mol Cell Biol, 1988 Jun, 8(6), 2361 - 6 Group II intron domain 5 facilitates a trans-splicing reaction; Jarrell KA et al.; A self-splicing group II intron of yeast mitochondrial DNA (aI5g) was divided within intron domain 4 to yield two RNAs that trans-spliced in vitro with associated trans-branching of excised intron fragments . Reformation of the domain 4 secondary structure was not necessary for the trans reaction, since domain 4 sequences were shown to be dispensable . Instead, the trans reaction depended on a previously unpredicted interaction between intron domain 5, the most highly conserved region of group II introns, and another region of the RNA . Domain 5 was shown to be essential for cleavage at the 5' splice site . It stimulated that cleavage when supplied as a trans-acting RNA containing only 42 nucleotides of intron sequence . The relevance of our findings to in vivo trans-splicing mechanisms is discussed. Ann Allergy, 1988 Jun, 60(6), 527 - 30 Ethanol-induced urticaria: a case report; Ting S et al.; Urticaria, after ingesting ethanol, is rare . A 36-year-old Caucasian male developed multiple, generalized, pruritic urticarial lesions 5 to 15 minutes after drinking alcoholic beverages of any type . A blinded challenge with 5 mL of chemically pure 95% ethanol in concentrated grape juice caused urticaria and an elevation of plasma histamine . Pure grape juice alone was unreactive . Prick skin tests with Brewers' yeast, ethanol, acetaldehyde, and acetic acid were negative . Hydroxyzine (25 mg, p.o., q.i.d.), given for three days prior to challenge, inhibited skin response and histamine release . Biopsy of the urticarial lesions caused by ethanol ingestion showed mast cell degranulation . In this subject, ethanol appeared to directly affect mast cell mediator release by non-IgE mechanisms. Microbiologia, 1988 Jun, 4(2), 87 - 96 {Induction of cellulases and xylanases in Aureobasidium pullulans}; Pou J et al.; The specificity or induction of wood-degrading enzymes using series of mono, di or polysaccharides as carbon source for the yeasts Geotrichum candidum and Trychosporon penicillatum and the yeast-like organism Aureobadidium pullulans were studied . The strain A . pullulans was the only one that when xylan or "steam exploded wood" were used as carbon source all the enzymes tested were found . This strain was unable to grow on carboxymethyl or Avicel cellulose . D-xylose was the nutritional inducer of beta-xylosidase and beta-xylanase but D-glucuronic acid induced CMCase activity and beta-glucosidase was produced with every carbon source . The 1,4 beta-xylobiose was not an inducer of beta-xylanase but with the structural 1,2-beta- and 1,3-beta-xylobiose isomers high levels of this enzyme were obtained. Sci Sin {B}, 1988 Jun, 31(6), 687 - 94 Chemical synthesis and cloning of secretin gene; Qian SW et al.; A DNA duplex coding for the 27 amino