Biochemistry, 1990 Jun 12, 29(23), 5413 - 22 Structure, biosynthesis, and function of glycosylphosphatidylinositols; Thomas JR et al.; The last few years have witnessed an explosion in our knowledge of GPI membrane anchors and related glycolipids and molecules where structure details are available, as illustrated in Figure 2 . There is now sufficient information on a handful of these molecules to allow a detailed comparison of their chemical structures . Despite a common structural theme, i.e., the presence of mannoglucosaminyl-PI, a great deal of diversity exists in both the glycan structures and the glycerol-linked aliphatic substituents . The complexities of the structures clearly show that a multitechnique approach is required in the elucidation of their structures . The anticipated publication of more structures from a wider range of organisms may reveal even greater diversity, as well as suggesting possible biosynthetic pathways . The details of a potential biosynthetic pathway in T . brucei are becoming apparent, but confirmation of its importance awaits the isolation and characterization of the enzymes involved . Leishmania, in which LPG, GPIs, GIPLs, and GPI membrane anchors are produced, may also provide an interesting system for biosynthetic studies . The recent description of a GPI biosynthetic system in yeast may provide the crucial breakthroughs necessary in unraveling the enzymes and sugar donors involved in the biosynthetic pathway and possibly the role of the GPI membrane anchor in the function of proteins containing these moieties . Knowledge of the solution structure (conformation), in addition to the complete chemical structure, of the T . brucei VSG anchor has led to speculation that the glycan fulfills a space-filling role in the VSG coat . Many other possible roles of GPI membrane anchors have been suggested, including the shedding and turnover of membrane proteins, signal transduction, and intracellular targeting . Nevertheless, the only function of GPIs that we can so far be certain of is that they anchor proteins or polysaccharide to a membrane . Regardless of the roles GPIs may or may not ultimately be shown to play, the fact that such a widely occurring structure has only recently been characterized serves as a reminder of the incompleteness of our knowledge of biological phenomena and the constant possibility of finding novel molecules in obvious places. Infect Immun, 1990 Jun, 58(6), 1538 - 44 Functional analysis of Histoplasma capsulatum-reactive T-cell hybridomas; Deepe GS Jr et al.; Activation of CD4+ T cells is a crucial step in the elimination of Histoplasma capsulatum yeast cells from tissues . However, only a limited amount of information exists concerning the immunobiology of H . capsulatum-reactive T cells that are CD4+ . To facilitate the analysis of the functional activities of this T-cell subpopulation, we developed a panel of 10 murine T-cell hybridomas from splenocytes of immune C57BL/6 mice . All hybridomas reacted with monoclonal anti-CD4+ antibody and released interleukin-2 after stimulation with histoplasmin . Within 3 weeks, the reactivity of hybridomas to histoplasmin declined dramatically, yet the cells responded vigorously to yeast-phase preparations that were enriched for cytosol, cell wall, or cell membrane . Of 10 hybridomas studied, only one recognized heterologous fungal antigens . Responsiveness to yeast-phase antigens was restricted by I-Ab . We mapped determinants in cytosol and cell wall or cell membrane by the technique of one-dimensional T-cell immunoblotting . The patterns of responses of hybridomas to cytosol were nearly uniform . All hybridomas responded to two immunodominant regions in cytosol with masses ranging from less than or equal to 18 to 26 kilodaltons (kDa) and 35 to 39 kDa . All hybridomas tested responded to determinants in the cell wall or cell membrane preparation with masses of 35 to 39 kDa . These hybridomas provide a useful tool for defining yeast-phase antigens that trigger T-cell activation. J Biol Chem, 1990 Jun 5, 265(16), 9314 - 8 Conversion of recombinant hirudin to the natural form by in vitro tyrosine sulfation . Differential substrate specificities of leech and bovine tyrosylprotein sulfotransferases; Niehrs C et al.; Hirudin, a tyrosine-sulfated protein secreted by the leech Hirudo medicinalis, is one of the most potent anticoagulants known . The hirudin cDNA has previously been cloned and has been expressed in yeast, but the resulting recombinant protein was found to be produced in the unsulfated form, which is known to have an at least 10 times lower affinity for thrombin than the naturally occurring tyrosine-sulfated hirudin . Here we describe the in vitro tyrosine sulfation of recombinant hirudin by leech and bovine tyrosylprotein sulfotransferase (TPST) . With both enzymes, in vitro sulfation of recombinant hirudin occurred at the physiological site (Tyr-63) and rendered the protein biochemically and biologically indistinguishable from natural hirudin . However, leech TPST had an over 20-fold lower apparent Km value for recombinant hirudin than bovine TPST . Further differences in the catalytic properties of leech and bovine TPSTs were observed when synthetic peptides were tested as substrates . Moreover, a synthetic peptide corresponding to the 9 carboxyl-terminal residues of hirudin (which include Tyr-63) was sulfated by leech TPST with a similar apparent Km value as full length hirudin, indicating that structural determinants residing in the immediate vicinity of Tyr-63 are sufficient for sulfation to occur. FEBS Lett, 1990 Jun 4, 265(1-2), 113 - 6 Copolymers of glutamic acid and tyrosine are potent inhibitors of oocyte casein kinase II; Tellez R et al.; Polypeptides rich in glutamic acid are strong inhibitors purified from isolated nuclei of Xenopus laevis oocytes of casein kinase II . The presence of tyrosine in these peptides greatly enhances their inhibitory capacity . Using casein as a substrate, copolyglu:tyr (4:1) has an I50 value of 20 nM, 250 fold lower than that of polyglutamic acid which is 5 microM . A similar large difference is observed when a synthetic peptide is used as substrate . The inhibition of copolyglu:tyr is competitive with casein and can be completely reversed by high ionic strength . The relative inhibitory capacity of the polypeptides tested, in descending order, is copolyglu:tyr (4:1) greater than copolyglu:tyr (1:1) greater than polyglu greater than copolyglu:phe (4:1) greater than copolyglu:ala (6:4) greater than copolyglu:leu (4:1) . The high affinity for tyrosine-containing acid peptides is shared by rat liver and yeast casein kinase II so that it seems to be a general property of these enzymes. Zhonghua Shen Jing Jing Shen Ke Za Zhi, 1990 Jun, 23(3), 133 - 5, 188 {Detection of erythrocyte immune function and circulatory immune complex in myasthenia gravis}; Guo F; The circulating immune complexes (CIC) and erythrocyte immune function in myasthenia gravis were studied . In order to examine CIC in the patients with myasthenia gravis the complement sensitized yeast cell agglutination (CSYCA) test and anti-C3-ELISA were used . The CIC positive rate was 92.3% in the patients tested . The CIC test was all negative in the normal control subjects . The levels of CIC were remarkable elevated in the patients with myasthenia gravis (P less than 0.01) . It was found that the rosette rate of red blood cell C3b receptor was 11.27 +/- 3.27% in the CIC negative patients with myasthenia gravis and 17.60 +/- 5.10% in CIC negative patients with myasthenia gravis . The erythrocyte immune function decreased remarkably in the CIC positive patients with myasthenia gravis (P less than 0.05) . The relationship between the decrease of the erythrocyte immune function and myasthenia gravis is discussed. Zhonghua Shen Jing Jing Shen Ke Za Zhi, 1990 Jun, 23(3), 130 - 2, 188 {Circulating immune complexes and myasthenia gravis (analysis of 78 cases)}; Tu LH; For the sake of studying the circulating immune complexes (CIC) of 78 patients with myasthenia gravis (MG), the agglutination assay of C3b sensitized with yeast cell (C3bSYCA) was used . The anti human C3 ELISA was also used for 21 out of the 78 patients . The positive results in the entire group of the patients studied were found to reach 82% and the IgG and IgM CIC were detected also with anti-human C3 ELISA . The titres of CIC thus determined decreased as the clinical conditions improved . It was considered that CIC might be regarded as a referential index to observe the patient's clinical condition and to evaluate the effect of treatment and to identify certain immune aberration of MG as well. Mycopathologia, 1990 Jun, 110(3), 125 - 32 Effect of peanut tannin extracts on growth of Aspergillus parasiticus and aflatoxin production; Azaizeh HA et al.; Twenty-three peanut (Arachis hypogaea L.) genotypes were evaluated for kernel resistance to Aspergillus parasiticus Spear . colonization and aflatoxin contamination when incubated under high relative humidity . Also, tannin-containing extracts from kernel coats (testae) and cotyledons of these genotypes were prepared and tested for their effect on A . parasiticus growth and aflatoxin production in vitro . The lowest degree of colonization, less than 30%, was noted in kernels from the genotypes, Toalson x UF 73-4022 (selections TX-798731 and TX-798736), A72118, SN 55-437, PI337409, and Florunner . Genotypes with low levels of colonization also had the lowest aflatoxin contamination . The coefficient of correlation between infection frequency and aflatoxin contamination was 0.66 . Higher levels of tannins were detected in the testae (23.9-97.2 mg g tissue) compared to the cotyledons (0.17-0.82 mg g tissue) . Some of the methanol-extracted and water-soluble tannin extracts from testae and cotyledons, when incorporated in yeast extract sucrose liquid medium (100 mg l), significantly inhibited A . parasiticus growth and reduced the levels of aflatoxin produced . There was no overall correlation between the peanut genotypes and the influence of tannin extracts on A parasiticus growth and aflatoxin production . However, correlations were higher for specific genotypes . For example, the coefficient of correlation between the ability of tannin extracts from testae of genotypes PI337409 and TX-798736 to inhibit aflatoxin production was 0.93 and 0.85 respectively. DNA Cell Biol, 1990 Jun, 9(5), 323 - 34 Cell-specific expression of a profilin gene family; Binette F et al.; Profilin is a ubiquitous eukaryotic protein that inhibits actin polymerization . We cloned and sequenced the two profilin genes from the acellular slime mold Physarum polycephalum . The genes, proA and proP, each contain two introns . Primer extension experiments showed two possible transcription start sites in the profilin A gene and one start site in the profilin P gene . The profilin A mRNA has two polyadenylation sites, which yield mRNAs of about 600 and 500 nucleotides . The profilin P mRNA has a single polyadenylation site . The protein sequences were deduced from cDNA nucleotide sequencing . The profilin A and profilin P proteins contain 125 amino acids and are 66% identical in sequence . They show sequence similarity to Acanthamoeba (approximately 54%), yeast (approximately 46%), mouse (approximately 22%), calf (approximately 21%), and human (approximately 21%) profilins . The presence of the profilin A and profilin P mRNAs was studied throughout the life cycle . Profilin A mRNA was found in amebas, encysted amebas, and mature spores . The profilin P mRNA was present in plasmodia and spherulating plasmodia . Most cell types of P . polycephalum contain either the amebal or the plasmodial profilin mRNA, and no cell contains both mRNAs . This is the first evidence for developmental regulation of profilin isoforms. Genitourin Med, 1990 Jun, 66(3), 193 - 9 Properties of Trichomonas vaginalis grown under chemostat controlled growth conditions; Lehker MW et al.; Trichomonas vaginalis isolates NYH 286 and IR 78 were grown in continuous flow culture conditions in a complex trypticase-yeast extract-maltose medium supplemented with heat-inactivated horse serum . Parasites could be stably maintained in the chemostat at high densities ranging from 1 x 10(6) to 1 x 10(7) organisms ml-1 . Growth densities, acid production, and profiles of total versus secreted trichomonad proteins were characterised at different rates of growth and pH . Growth rate influenced the extent of parasite production of acid and the shedding of proteins into the medium but had no effect on overall parasite density . Lowering the pH from 6.0 to 5.0 resulted both in a decrease of cell density and acid production . At pH 4.5 isolate IR 78 but not NYH 286 was capable of growth and multiplication, showing the ability of some isolates to survive at the vaginal pH of healthy individuals . At this lower pH, however, isolate NYH 286 but not IR 78 synthesised new proteins which were detectable in stained gels . Also, inoculation of the chemostat with isolate NYH 286 comprising a mixture of fluorescent (positive, pos) and non-fluorescent (negative, neg) trichomonads as defined by monoclonal antibody reactivity to a surface immunogen resulted in a change in the parasite population to an almost homogeneous neg phenotype . These neg phenotype organisms switched back to pos phenotype after transfer to test tubes. Vaccine, 1990 Jun, 8(3), 192 - 8 Protective efficacy of a novel hepatitis B vaccine consisting of M (pre-S2 + S) protein particles (a third generation vaccine); Fujisawa Y et al.; The protective efficacy of a new type of yeast-derived hepatitis B (HB) vaccine (TGP-943, subtype adr), which was formulated from modified M (pre-S2 + S; P31) protein (M-P31c) particles, was investigated in chimpanzees . Animals were injected intramuscularly three times at 4-week intervals with doses of 10 or 40 micrograms (as a protein) of TGP-943 . There were no significant differences in the immunogenicity of 10 micrograms compared to that of 40 micrograms of TGP-943 in terms of anti-S antibody response, while the induction and persistence of anti-pre-S2 antibodies seemed dose-related . Chimpanzees, vaccinated with 40 micrograms of TGP-943, produced anti-pre-S2 antibodies 2 weeks after the first injection, which appeared earlier than anti-HBs (S) antibodies . A maximum level of the anti-pre-S2 antibodies was reached 2 weeks after the second injection . Apart from immunization with TGP-943, chimpanzees injected with denatured TGP-943, consisting of 10 micrograms (as a protein) of non-particulate M-P31c antigen, produced anti-pre-S2 antibodies with a non-protecting level of anti-S antibodies (less than 10 mIU ml-1) . Five weeks after the third injection, all animals were challenged intravenously with 1000 chimpanzee infectious units of HBV subtype (ayw) and were protected as confirmed by normal serological markers, no signs of infection in the sera and liver biopsies, and no detection of HBV-DNA by PCR method . No side effects from inoculation with TGP-943 or denatured TGP-943 were also encountered in any animals. Arch Biochem Biophys, 1990 Jun, 279(2), 298 - 304 Carbohydrate-binding specificity of the daffodil (Narcissus pseudonarcissus) and amaryllis (Hippeastrum hybr.) bulb lectins; Kaku H et al.; The carbohydrate binding specificity of the daffodil (Narcissus pseudonarcissus; NPA) and amaryllis (Hippeastrum hybr.; HHA) lectins, isolated from extracts of their bulbs by affinity chromatography on immobilized mannose, was studied by quantitative precipitation, sugar hapten inhibition, and affinity chromatography on the immobilized lectins . These lectins gave strong precipitation reactions with several yeast mannans, but did not precipitate with alpha-D-glucans (e.g., dextrans and glycogen) . Interestingly, both lectins reacted strongly with yeast galactomannans having multiple nonreducing terminal alpha-D-galactosyl groups, a synthetic linear alpha-1,6-mannan, and an alpha-1,3-mannan (DP = 30) . Treatment of the linear alpha-1,3-mannan with periodate, resulting in oxidation of the terminal, nonreducing mannosyl group, did not reduce its reactivity with NPA or HHA . Taken together, these observations suggest that NPA and HHA react not only with terminal but also with internal alpha-D-mannosyl residues . Sugar hapten inhibition studies showed these lectins to possess the greatest specific activity for alpha-D-mannosyl units whereas D-Glc and D-GlcNAc did not inhibit either lectin precipitation system . Of the oligosaccharides tested, the best inhibitor of NPA interaction was alpha-1,6-linked mannotriose, which was twice as good an inhibitor as Man alpha 1,6Man alpha-O-Me and 10 times better than methyl alpha-D-mannoside . On the other hand, oligosaccharides containing either 1,3- or 1,6-linked mannosyl units were good inhibitors of the HHA-mannan precipitation system (6- to 20-fold more active than D-Man) . These results indicate that both lectins appear to possess an extended binding site(s) complementary to at least three 1,6-linked alpha-mannosyl units . Various glycosylasparagine glycopeptides which contain alpha-1,6-Man units were retarded on the immobilized NPA column . On the other hand, those containing either alpha-1,3- or alpha-1,6-mannosyl residues were retarded on the immobilized HHA column; Man5-GlcNAc2-Asn {containing two Man alpha 1,3(Man alpha 1,6) units} bound to the HHA column . On the contrary, glycopeptides with hybrid type glycan chains were not retarded on either column . These two new lectins which differ in their fine sugar binding specificity from each other, and also from the snowdrop lectin, should prove to be useful probes for the detection and preliminary characterization of glycoconjugates on cell surfaces and in solution. Int J Artif Organs, 1990 Jun, 13(6), 359 - 64 Influence of membranes on generation of beta 2 M and release of leukocyte lysosomal enzymes; Stein G et al.; Normal leukocyte functional capacity was investigated by evaluation of phagocytosis of opsonised yeast cells in a radiometric test system . After incubation with dialysis membranes (different cellulosic membranes, polysulfon membrane (PS), polymethylmetacrylate membrane (PMMN), the phagocytosis index, expressed as percent decrease with respect to initial values without membrane, decreased by 10%-25% . The most pronounced effect was observed with PS, cuprophane, modified cellulose and PMMA . The results are not related to differences in the viability of PMN during the test procedure; dead PMN amounted to about 4-6.5% . A significant increase in beta-NAG and beta-Gluc activities was released in the supernatants of the phagocytosis suspensions . This increase activity can be explained by the phagocytosis of PMN but it was not influenced by membrane contact . There was no influence of membrane contact or phagocytosis activity of PMN on the beta 2 M concentration in the supernatant demonstrating that no in vitro generation during incubation with either membrane exists. Gan To Kagaku Ryoho, 1990 Jun, 17(6), 1095 - 103 {Progress of research on xeroderma pigmentosum}; Tanaka K; Xeroderma pigmentosum (XP) is an autosomal recessive human disease, clinically characterized by high incidence of skin cancers on sun-exposed areas . XP cells are hypersensitive to killing by ultraviolet light (UV), because they have a defect in DNA excision repair of UV-induced DNA damages . Genetic complementation analysis by cell fusion has identified 9 genetic complementation groups, designated groups A through H and a variant . However, the genetic basis of the physiological defect of XP has not yet been characterized . Recently, XP genes and human DNA repair genes have been molecularly cloned by DNA transfection methods . Molecular biological analysis of these genes should be a clue to elucidating the molecular mechanism of DNA repair in human . Moreover, an in vivo microinjection system and an in vitro system for study of DNA repair synthesis promoted by human cell extract have been developed and they can be utilized as assays during the purification of protein factors that complement repair defective XP cells . A nuclear factor that binds to DNA lesion has been identified and it was defective in group E XP cells . Yeast homolog of this nuclear factor appears to be a photolyase. Biochem J, 1990 Jun 1, 268(2), 521 - 4 The structure of the human gene encoding protein gene product 9.5 (PGP9.5), a neuron-specific ubiquitin C-terminal hydrolase; Day IN et al.; Database search using a bovine thymus ubiquitin C-terminal hydrolase sequence indicated 54% sequence identity with the abundant human neuron-specific protein gene product 9.5 (PGP9.5), which was then shown to possess the same activity {Wilkinson, Lee, Deshpande, Duerksen-Hughes, Boss & Pohl (1989) Science 246, 670-673} . A yeast counterpart of the enzyme is also known . The human PGP9.5 gene, described here, spans 10 kb, contains nine exons and displays 5' features some common to many genes and some common with neurofilament neuron-specific enolase and Thy-1-antigen gene 5' regions. Mol Gen Genet, 1990 Jun, 222(1), 120 - 8 Nucleotide sequence and analysis of NMR, a negative-acting regulatory gene in the nitrogen circuit of Neurospora crassa; Young JL et al.; In Neurospora the expression of a set of unlinked structural genes, which allows utilization of various nitrogen-containing compounds, is controlled by the positive-acting nit-2 gene and the negative-acting nmr gene . The nucleotide sequence of the nmr gene has been determined and a long open reading frame which encodes a putative protein of 54854 daltons has been identified . A full-length cDNA clone was obtained and its the sequence revealed that the nmr gene contains no introns . The transcriptional start and stop sites have been mapped by S1 nuclease and primer extension . Site-directed mutagenesis was used to introduce stop codons at various locations in the nmr coding region . Transformation assays showed that the proteins lacking up to 16% of the carboxyl-terminus were still functional . Homology searches showed that the nmr protein is homologous to the yeast arginine regulatory gene AR-GRII. Zhong Xi Yi Jie He Za Zhi, 1990 Jun, 10(6), 345 - 7, 325 {Relation of tongue color changes and red-cell immune adherence activity in patients with malignant bone tumor}; Shen L; This article presents that preliminary study on the changes of the tongue colour and the red-cell immune adherence (RCIA) activity in 40 patients with malignant bone tumor, as compared with 40 healthy persons . The results showed that the rate of erythrocyte C3b receptor yeast rosette were no statistically significant different between the pink and the crimson tongues in the bone tumor group and control group (P greater than 0.05) . It appeared RCIA activity was no depression in the patients with malignant bone tumor during pink and crimson tongue . In the bone tumor group with the pale and the cyanotic tongues, the rate of erythrocyte C3b receptor yeast rosette and erythrocyte immune complex rosette was significantly lower than that of the control group (P less than 0.05, P less than 0.01) . It appeared that RCIA activity in these patients was markedly impaired . The authors suggested that the tongue colour changes in the patients with malignant bone tumors could roughly reflect the level of RCIA activity . This phenomenon is beneficially to predict the body capacity against the malignant tumor. Mol Biochem Parasitol, 1990 Jun, 41(1), 7 - 15 A Trypanosoma brucei small RNP particle containing the 5S rRNA; Michaeli S et al.; In mammalian cells, approximately 50% of the 5S rRNA is found in ribosomes, and the remainder in a small particle, the 5S rRNA/ribosomal protein L5 complex, which is thought to be a precursor in ribosome assembly . Trypanosoma brucei, an African trypanosome, is one of the most primitive eukaryotic organisms which have been studied, and it likewise possesses a 5S rRNA species, a small proportion of which is found in an apparent ribonucleoprotein-(RNP) complex . Like the mammalian RNP particle, the T . brucei particle has a sedimentation coefficient of about 7S in sucrose gradients; unlike its mammalian counterpart, the complex is not disrupted by high salt and can be fractionated in cesium sulfate density gradients at a density characteristic of RNP complexes (1.45 g ml-1) . Our studies demonstrate the the T . brucei 7S RNP contains 5S rRNA in association with a 36-kDa rRNA binding protein which not only shares molecular size, but also immunological determinants, with the yeast ribosomal protein YL3, and its mammalian homologue, L5 . These results indicate that the RNP complex formed between the 5S rRNA and the 36-kDa ribosomal protein is conserved throughout great evolutionary distances between eukaryotic species. J Am Acad Dermatol, 1990 Jun, 22(6 Pt 1), 993 - 8 The in vitro antifungal activity of ketoconazole, zinc pyrithione, and selenium sulfide against Pityrosporum and their efficacy as a shampoo in the treatment of experimental pityrosporosis in guinea pigs; Van Cutsem J et al.; The fungistatic and fungicidal activity of ketoconazole, zinc pyrithione, and selenium sulfide against Pityrosporum, a yeast thought to play a pathogenic role in seborrheic dermatitis and dandruff, was assessed in Dixon broth for Pityrosporum ovale and Sabouraud broth for Pityrosporum pachydermatis . Ketoconazole inhibited growth at concentrations ranging from 0.001 to 1 micrograms/ml . For zinc pyrithione and selenium sulfide higher concentrations were needed . In a guinea pig model the efficacy of treatment with four shampoos (Nizoral {Jansen}, EDS Zinc {Schering}, Zinkan {Lederle}, and Selsun {Abbott}) was compared . The animals were inoculated for 7 consecutive days on intact skin . The lesions were scored for erythema, folliculitis, and hyperkeratosis 24 hours after the last inoculation and after treatment . Final evaluations were made 13 days after infection (10 days after last shampoo application) . Treatment with undiluted and diluted (1:10) shampoos showed consistently superior clinical and mycologic results for Nizoral shampoo . None of the shampoos produced side effects. New Biol, 1990 Jun, 2(6), 505 - 11 The molecular control of transport vesicle fusion; Wattenberg BW; The fusion of transport vesicles with the appropriate target membrane in constitutive transport is a complex and well-controlled process . Many of the molecular details of the reactions that result in this control are being revealed through the use of cell-free assays of protein transport as well as by the study of the molecular genetics of secretion in yeast . Kinetic analyses have indicated that several structural intermediates are formed after transport vesicles attach to their destination, but before they fuse with the appropriate membrane . Proteins that mediate the formation and processing of these intermediates have been identified . Included among these are small molecular weight GTP-binding proteins . This intricate set of reactions may ensure the fidelity of transport and guard the integrity of the organelles along the transport pathway. Bioessays, 1990 Jun, 12(6), 253 - 8 Molecular machinery required for protein transport from the endoplasmic reticulum to the Golgi complex; Hicke L et al.; The cellular machinery responsible for conveying proteins between the endoplasmic reticulum and the Golgi is being investigated using genetics and biochemistry . A role for vesicles in mediating protein traffic between the ER and the Golgi has been established by characterizing yeast mutants defective in this process, and by using recently developed cell-free assays that measure ER to Golgi transport . These tools have also allowed the identification of several proteins crucial to intracellular protein trafficking . The characterization and possible functions of several GTP-binding proteins, peripheral membrane proteins, and an integral membrane protein during ER to Golgi transport are discussed here. Mol Cell Biol, 1990 Jun, 10(6), 2848 - 54 A genomic clone encoding a novel proliferation-dependent histone H2A.1 mRNA enriched in the poly(A)+ fraction; Fecker L et al.; Replication-dependent histone mRNAs are prime examples of nonpolyadenylated mRNAs . We isolated and characterized cDNAs and a genomic clone for a replication-dependent histone H2A.1 mRNA which segregated into the poly(A)+ fraction during mRNA isolation through an oligo(dT)-cellulose column . However, the results of sequencing of the genomic clone suggested that the mRNA did not contain a poly(A) tail . Instead, the genomic sequence revealed a nonterminal oligo(A) tract directly upstream from the typical 3'-terminal hairpin loop of replication-dependent histone mRNAs . The nonterminal oligo(A) tract consisted of 14 adenylate residues interrupted by one guanylate residue (A4GA10) . We concluded that this short oligo(A) stretch mediated binding of the mRNA to oligo(dT) even after stringent washes with 0.1 M NaCl, indicating that rather short oligo(A) sequences can ensure binding to oligo(dT)-cellulose . The cDNA and genomic clones contained an AAATAAG sequence at the end of the coding region . It has been suggested that this sequence contains a polyadenylation signal in some yeast and mouse transcripts, but it does not function as a polyadenylation signal in the histone transcript described in this paper. Nippon Juigaku Zasshi, 1990 Jun, 52(3), 527 - 31 A new staining solution for the morphological studies of fungi and Prototheca; Pal M et al.; A new staining solution named "PHOL" is described for studying the morphology of clinical and environmental isolates of fungi and Prototheca . In total 69 isolates were used for assessing the efficacy of this newly formulated stain . The solution has shown good ability to stain all 62 isolates of fungi and 7 of Prototheca, and thus helped in the identification of the organisms . The new solution has the potential to stain the young as well as old isolates of fungi and Prototheca . It is emphasized that a wider application of this newly introduced inexpensive and safe staining solution will help the mycologists to study the morphological characteristics of yeast, molds, and Prototheca obtained from man, animals and environment. Nature, 1990 May 31, 345(6274), 458 - 60 Telomeres shorten during ageing of human fibroblasts; Harley CB et al.; The terminus of a DNA helix has been called its Achilles' heel . Thus to prevent possible incomplete replication and instability of the termini of linear DNA, eukaryotic chromosomes end in characteristic repetitive DNA sequences within specialized structures called telomeres . In immortal cells, loss of telomeric DNA due to degradation or incomplete replication is apparently balanced by telomere elongation, which may involve de novo synthesis of additional repeats by novel DNA polymerase called telomerase . Such a polymerase has been recently detected in HeLa cells . It has been proposed that the finite doubling capacity of normal mammalian cells is due to a loss of telomeric DNA and eventual deletion of essential sequences . In yeast, the est1 mutation causes gradual loss of telomeric DNA and eventual cell death mimicking senescence in higher eukaryotic cells . Here, we show that the amount and length of telomeric DNA in human fibroblasts does in fact decrease as a function of serial passage during ageing in vitro and possibly in vivo . It is not known whether this loss of DNA has a causal role in senescence. J Immunol Methods, 1990 May 25, 129(2), 283 - 90 A new photometric microassay for the quantitation of macrophage Fc receptor function . In vitro enzyme-containing immune complexes clearance (EIC) assay; Matsumoto T et al.; A new photometric microassay for immune complex (IC) binding to macrophages was developed in a homologous system using glucose oxidase-anti-glucose oxidase complexes (GOAGO) as a model for IC clearance in vitro . Thioglycollate-elicited murine peritoneal cells were incubated with GOAGO solution and then cell-associated glucose oxidase activity was measured after the washing and solubilization of the cell membrane in a microtitre plate . GOAGO binding to macrophages was inhibited in the presence of either IgG or its Fc fragments in a dose-dependent manner, while yeast mannan or IgG Fab fragments had no effect . These results indicated that this binding occurred solely via the Fc receptors on the macrophages . The Fc receptors for GOAGO were eliminated by trypsin digestion of the cells . When the macrophages were cultured with LPS or TPA, GOAGO binding was enhanced compared to that of control, whereas carrageenan treatment suppressed GOAGO binding . The present results suggest that this assay may be of value in the measurement of IC clearance and for studying the expression of Fc receptors on macrophages. J Biol Chem, 1990 May 15, 265(14), 8159 - 63 Common protein-binding sites in the 5'-flanking regions of human genes for cytochrome c1 and ubiquinone-binding protein; Suzuki H et al.; The single nuclear gene encoding human ubiquinone-binding protein, a subunit of the cytochrome bc1 complex, was isolated previously (Suzuki, H., Hosokawa, Y., Toda, H., Nishikimi, M., and Ozawa, T . (1989) Biochem . Biophys . Res . Commun . 161, 371-378) . The 5'-flanking region contains four putative CCAAT boxes, one putative NF-Y-binding site, and one sequence homologous to the AP-1 recognition site, but lacks typical TATA and GC boxes . Three common nucleotide sequences (5'-TATTCAGGT-3', 5'-ATCTGGCT-3', and 5'-TGGTGA(T/G)AG-3', designated Mt1, Mt3, and Mt4, respectively) were found in the 5'-flanking regions of the genes for human ubiquinone-binding protein and for human cytochrome c1, another subunit of the cytochrome bc1 complex . All three sequences are located in the 280-base pair BamHI-HindIII fragment of the ubiquinone-binding protein gene and in the 154-base pair SalI-PstI fragment of the cytochrome c1 gene . Interestingly, a computer-assisted homology search revealed that the SalI-PstI fragment of the cytochrome c1 gene is related to the long terminal repeat of an endogenous retrovirus . Sequences highly homologous to the three Mt-sequences were also found in the 5'-flanking regions of the genes for the beta subunit of human F0F1-ATPase and rat somatic cytochrome c . The Mt1 sequence (TATT-CAGGT) is similar to the GFII recognition site (RTCACGTG) found in the 5'-flanking regions of the yeast nuclear genes for three cytochrome bc1 complex subunits . Gel retardation assay demonstrated that protein factors in a whole HeLa cell extract bound to both of those fragments . At least three different specific binding sites were thought to exist in the fragments . Specific binding of the protein factors to the sites, possibly to the three Mt sequences, may play an important role in the coordinate regulation of the transcription of nuclear genes encoding subunits responsible for mitochondrial oxidative phosphorylation. Anal Biochem, 1990 May 15, 187(1), 104 - 8 Enzymatic synthesis of UDP-GlcN by a two step hollow fiber enzyme reactor system; Ropp PA et al.; UDP-GlcN was synthesized from GlcN and UTP by a two step hollow fiber enzyme reactor method . In step 1, GlcN was converted to GlcN 6-P and then to GlcN 1-P by hexokinase and phosphoglucomutase, respectively, and UTP was used as the phosphate donor . In step 2, GlcN 1-P was converted to UDP-GlcN by UDP glucose pyrophosphorylase . All the enzymes required for the synthesis of UDP-GlcN were enclosed in hollow fiber bundles which allow for the free diffusion of substrates and products across the membranes to and from the enzymes, allow for the reutilization of the enzymes, and simplify the isolation of the product, UDP-GlcN . We show that both UTP and GlcN 6-P are inhibitors of the yeast UDPG pyrophosphorylase and therefore their concentrations must be regulated to obtain maximum yields of UDP-GlcN . The UDP-GlcN produced can be N-acetylated with {14C}acetic anhydride to produce UDP-{14C}GlcNAc . This method can also be used to synthesize {32P}UDP-GlcN and {32P}UDP-GlcNAc from {alpha-32P}UTP and GlcN 1-P. Drug Metab Dispos, 1990 May-Jun, 18(3), 276 - 80 Ethyl carbamate metabolism: in vivo inhibitors and in vitro enzymatic systems; Yamamoto T et al.; The metabolism of ethyl carbamate and the localization of its metabolites have been shown to be almost completely inhibited by ethanol in the mouse {Waddell, Marlowe, Pierce: Food Chem . Toxicol.25, 527 (1987); Yamamoto, Pierce, Hurst, Chen, Waddell: Drug Metab . Dispos . 16, 355 (1988)} . The enzyme system catalyzing this metabolism which is inhibited by ethanol now has been further investigated in both in vivo and in vitro studies . There is a direct, highly significant relationship between the extent of metabolism of ethyl carbamate and covalent binding of metabolites to liver protein . Paraoxon, carbaryl, CCl4 ethanol, methimazole, 4-methylpyrazole, diethyl maleate, ethyl N-hydroxycarbamate, and t-butyl carbamate inhibit, to different extents, the metabolism of ethyl carbamate in vivo; SKF-525A, CoCl2, Cacyanamide, chloral hydrate, 2-oxo-4-thiazolidine carboxylic acid, allopurinol, and methyl carbamate do not . Porcine liver esterase, yeast aldehyde dehydrogenase and mouse liver catalase catalyzed the metabolism in vitro; dog or bovine catalase, acid phosphatase, alcohol dehydrogenase, or carbonic anhydrase did not under the conditions tested . Paraoxon, 4-methylpyrazole, carbaryl, and NaF significantly inhibited the hydrolytic activity of mouse liver homogenates toward p-nitrophenyl acetate; ethanol or ethyl carbamate did not . However, each of these, except 4-methylpyrazole, inhibited the metabolism of ethyl carbamate by mouse liver homogenate or porcine liver esterase to about the same extent . Ion exchange chromatography of mouse liver cytosol revealed that the fraction with ability to metabolize ethyl carbamate co-chromatographed almost exactly with the ability to hydrolyze p-nitrophenyl acetate . It is proposed that ethyl carbamate is metabolized in the mouse, at least partially, by esterases; however, metabolism by other enzyme systems cannot be excluded. Science, 1990 May 11, 248(4956), 732 - 5 Organization of the human and mouse low-affinity Fc gamma R genes: duplication and recombination; Qiu WQ et al.; Receptors for immunoglobulin G immune complexes (Fc gamma RII and Fc gamma RIII) are expressed on most hematopoietic cells and show much structural and functional diversity . In order to determine the genetic basis for this diversity, a family of genes encoding the human and mouse receptors was isolated and characterized . Humans have five distinct genes for low-affinity Fc gamma Rs, in contrast to two in the mouse . With the use of yeast artificial chromosomes, the genes encoding the human receptors were oriented and linked, which established the structure of this complex locus . Comparison of the human and mouse genes generated a model for the evolutionary amplification of this locus. Science, 1990 May 11, 248(4956), 727 - 30 Cloning of a 67-kD neutrophil oxidase factor with similarity to a noncatalytic region of p60c-src; Leto TL et al.; Chronic granulomatous diseases (CGDs) are characterized by recurrent infections resulting from impaired superoxide production by a phagocytic cell, nicotinamide adenine dinucleotide phosphate (reduced) (NADPH) oxidase . Complementary DNAs were cloned that encode the 67-kilodalton (kD) cytosolic oxidase factor (p67), which is deficient in 5% of CGD patients . Recombinant p67 (r-p67) partially restored NADPH oxidase activity to p67-deficient neutrophil cytosol from these patients . The p67 cDNA encodes a 526-amino acid protein with acidic middle and carboxyl-terminal domains that are similar to a sequence motif found in the noncatalytic domain of src-related tyrosine kinases . This motif was recently noted in phospholipase C-gamma, nonerythroid alpha-spectrin (fodrin), p21ras-guanosine triphophatase-activating protein (GAP), myosin-1 isoforms, yeast proteins cdc-25 and fus-1, and the 47-kD phagocyte oxidase factor (p47), which suggests the possibility of common regulatory features. J Biol Chem, 1990 May 5, 265(13), 7687 - 92 Structural organization of the rat cytochrome c oxidase subunit IV gene; Yamada M et al.; We have isolated two overlapping clones covering the entire length of the gene of nuclear-encoded sub-unit IV of cytochrome c oxidase (COXIV) from a rat genomic library in Charon 4A and determined the structural organization of the gene . The gene spans approximately 7.0 kilobases and contains five exons interrupted by four introns . Of these exons, exon 2 codes for a whole length of the presequence of the rat COXIV precursor protein, while exons 3 to 5 encode three distinct structural domains of the mature protein . The 5'-flanking region of the gene lacks conventional TATA and CAAT boxes, but has a high G + C content and contains two putative binding sites for transcription factor SP1 and a sequence resembling the AP-4 responsive element . These results indicate that the promoter region of the rat COXIV gene possesses characteristic features common in housekeeping genes . The chloramphenicol acetyltransferase assay performed by constructing an improved phagemid, pBlueCAT3, revealed that a 773-base pair DNA fragment immediately preceding the cap site has a strong promoter activity . An octanucleotide sequence, -TTCTTGGT-, which is very close to the yeast HAP2/HAP3 responsive element, is located in the 5'-upstream region of the present gene. Mycopathologia, 1990 May, 110(2), 93 - 9 Ochratoxinogenicity of Aspergillus ochraceus strains from nephropathic and non-nephropathic areas in Yugoslavia; Cvetnic Z et al.; Mycological analyses of 855 samples of stored grains and dried meat collected in period 1980-1987 from individual households in the nephropathic and wider non-nephropathic area in SR Croatia in Yugoslavia showed 10% of samples to be contaminated with Aspergillus ochraceus . Ability to produce ochratoxin A (OA) was tested in 70 samples (27 from nephropathic areas and 43 from non-nephropathic areas) . The detection was carried out under UV-light (365 nm) (light blue fluorescence) and 6 OA-producers were found . A biosynthetic procedure on liquid nutritional substrate with saccharose and yeast extract as well as a method using wet crushed wheat revealed that 37% of the samples from a nephropathic area, and 35% of the samples from a non-nephropathic area produce OA . In the nephropathic area 1/10 strains was a strong producer of OA (concentration crushed wheat 135 mg/kg, and 240 mg/l on YES liquid substrate), 1/10 strains was a moderate producer (concentration 16.6 mg/l and 0.07-7.0 mg/l and 0.1-10.4 mg/kg) . Among the strains isolated from a wider non-nephropathic area no strong producers of OA were found, but 2/15 strains were moderate producers of OA (concentration of OA 20.4-27.0 mg/l and 15.0-33.7 mg/kg) . The other strains, 8/10 on the crushed wheat and 13/15 on the liquid substrate, were weak producers of OA with concentrations of OA between 0.2-9.0 mg/l and 0.2-10.0 mg/kg with the two methods respectively. Can J Public Health, 1990 May-Jun, 81(3), 235 - 6 Study of the abnormal cervical-vaginal cytology of sexually active young women living within the Waterloo region; Redmond M et al.; PIP: 618 women aged 13-36 attending a family planning clinic took part in a sexual health survey during 1985 and 1986 . They were followed up for one year in order to ascertain whether an increase in atypical and dysplastic pap smears was linked to a certain lifestyle . The routine examination included a pap smear, a cervical swab for gonorrhea and other vaginal flora . Cultures for herpes and chlamydia were done . Benign atypia infection was treated followed by repeat pap smears 6-8 weeks and 2-3 months later . 7% of the women were virgins, 72% had smoked, and 62% presently smoked (an average of 9 cigarettes daily) . Average age at first intercourse, was 16, and partners ranged from 1 to 35 . Condom use was most prevalent: 15% used it "always", 20% "often", and 36% "occasionally" . The oral contraceptive use pattern was: 60% "always" used it and 10% "often" relied on it . 1/4th used the rhythm method . 90% had never been pregnant, 9% had, and 1% were unsure . of 49 pregnancies 41 ended in abortion . After examining 581, 4 cases condylomata and 1 case of genital herpes were found . 60% of the women had normal cervical cytology, 32% showed atypia, and 8% displayed dysplasia . 22% of the atypias were linked to inflammation and 6% were related to yeast . 9% of repeat atypia or dysplasia was less severe by biopsy, while 54% turned out to be more severe than the pap smear results, although they tend to indicate more severe changes . 8% showed less severe cytology, and 59% more severe cytological changes . 78% of the group with mild dysplasia had smoked, as opposed to 22% that had "never smokes" . Those with mild dysplasia were 4 times more likely to smoke at the time of their initial visit (79%) than nonsmokers (21%) . 54% of those with normal pap smear reported "often" or "always" using condoms, whereas only 10% of the group with dysplasia reported similar use pattern . These findings uphold the practice of following the repeated atypical, or initial dysplastic pap smear results with colposcopy and biopsy, especially within this subgroup of the population . J Clin Microbiol, 1990 May, 28(5), 1054 - 6 Enzymatic activity profiling as a potential biotyping method for Ajellomyces dermatitidis; Summerbell RC et al.; Enzyme profiling was investigated as a means of recognizing biotypes and individual strains of Ajellomyces dermatitidis (anamorph, Blastomyces dermatitidis) . Eighteen North American and 2 African representatives were tested with the Yeast-IDENT enzymatic activity profiling system (Analytab Products, Plainview, N.Y.) . Significant variation was found between isolates, particularly in beta-galactosaminidase activity. Mycoses, 1990 May, 33(5), 247 - 50 Malassezia pachydermatis isolation from a scarlet macaw; Breuer-Strosberg R et al.; A repeated isolation of Malassezia pachydermatis Weidman from a scarlet macaw is reported . This is the first report of birds infected with this yeast. J Nat Prod, 1990 May-Jun, 53(3), 609 - 14 7-deoxy-6-epi-castanospermine, a trihydroxyindolizidine alkaloid glycosidase inhibitor from Castanospermum australe; Molyneux RJ et al.; A new indolizidine alkaloid has been isolated from the seeds of Castanospermum {1} australe and identified as 7-deoxy-6-epi-castanospermine by ms and 1H- and 13C-nmr spectroscopy . The alkaloid is the first trihydroxylated indolizidine to be isolated from this plant and may represent an intermediate in the biosynthetic pathway to the tetrahydroxy-indolizidines and -pyrrolizidines . It inhibits amyloglucosidase and yeast alpha-glucosidase but is significantly less active as a glycosidase inhibitor than its isomer swainsonine {2} and the tetrahydroxylated alkaloids castanospermine {3}, 6-epi-castanospermine {4}, and australine {5}. Am J Emerg Med, 1990 May, 8(3), 216 - 9 In the shadow of the Baron: sudden death due to Munchausen syndrome; Nichols GR 2nd et al.; The authors describe a woman who died suddenly in an emergency department restroom after self-injection of corn starch in an attempt to gain admission to the hospital . At autopsy, multiple pulmonary vascular corn starch granulomata were identified, as well as pulmonary arteries filled with brewer's yeast . Retrospective evaluation of the patient's medical history showed multiple signs of Munchausen Syndrome . The authors suggest that the case illustrates the importance of scene investigation and review of the medical history by emergency department personnel and the forensic pathologist. Thorax, 1990 May, 45(5), 362 - 8 Effects of an inhaled corticosteroid, budesonide, on alveolar macrophage function in smokers; Bergstrand H et al.; Selected functions of alveolar macrophages obtained by bronchoalveolar lavage of 12 healthy smokers were examined before and after eight weeks' treatment with an inhaled glucocorticosteroid, budesonide (400 micrograms twice daily) . After budesonide treatment spontaneous as well as opsonised zymosan triggered prostaglandin E2 (PGE2) secretion from harvested cells was reduced; no such reduction in opsonised zymosan triggered leukotriene B4 (LTB4) production was observed . Neither the capacity to phagocytose opsonised yeast particles nor the superoxide radical generation triggered by the calcium ionophore A23187, 4 beta-phorbol 12-myristate 13-acetate (PMA), or opsonised zymosan ex vivo were more than marginally affected by the glucocorticosteroid treatment in vivo . Lavage fluid concentrations of angiotensin converting enzyme (ACE), however, after treatment were twice those before treatment and concentrations of fibronectin were reduced to half . Albumin concentrations in lavage fluid were not affected by the glucocorticosteroid treatment . In separate experiments treatment of alveolar macrophages with 10(-7) or 10(-6) M budesonide overnight in vitro did not affect their superoxide radical or PGE2 generation but significantly blocked LTB4 release . These data indicate that inhaled gluco-corticosteroid treatment may affect synthesis or release (or both) of ACE and fibronectin by alveolar macrophages from healthy smokers whereas other functions of these cells, such as the generation of reactive oxygen derived products ex vivo, are only marginally affected. J Infect Dis, 1990 May, 161(5), 865 - 8 Simultaneous vaccination for hepatitis A and B; Flehmig B et al.; Seronegative volunteers (15) were vaccinated at three 1-month intervals with a combined hepatitis A and B vaccine . The vaccine contained a killed hepatitis A vaccine made from hepatitis A virus (HAV) propagated in diploid human fibroblast cell cultures and hepatitis B surface antigen (HBsAg) produced in yeast . After only one injection, all volunteers developed neutralizing antibodies for HAV with antibody titers comparable to those found after gamma globulin administration . Compared with the antibody titers in sera of volunteers vaccinated with HAV vaccine alone, the antibody titers against HAV were significantly higher in the sera of the volunteers simultaneously vaccinated against hepatitis B virus (HBV) after three vaccinations . Of 15 volunteers, 14 seroconverted against HBV after three injections . In 11 volunteers tested 48 weeks after vaccination, antibody titers against HAV and HBV remained high. New Biol, 1990 May, 2(5), 389 - 401 p34cdc2: the S and M kinase? Pines J, Hunter T. In the yeast cell cycle, the induction of two very different processes, DNA synthesis (S-phase) and mitosis (M-phase), requires the same serine/threonine-specific protein kinase p34cdc2, which has been highly conserved through evolution . On the basis of work conducted largely in multicellular eukaryotes, it has recently been suggested that p34cdc2 is able to perform these two mutually exclusive roles by phosphorylating different sets of substrates through a cell cycle-dependent association with other proteins that dictate the substrate specificity of the protein kinase . To recognize its mitotic substrates, p34cdc2 associates with one of the cyclins--a family of proteins of two distinct but related types (A and B) characterized by their periodic destruction at each mitosis . In interphase, the formation of a complex between p34cdc2 and another protein (or proteins) would allow the phosphorylation of a different set of proteins involved in the G1 to S transition . This review focuses on the evidence for this appealing simple model and the nature of the putative substrates proposed. APMIS, 1990 May, 98(5), 415 - 22 Different cell surface and phagocytic properties in mononuclear phagocytes from blood and alveoli . A comparative study of blood monocytes and alveolar macrophages from human nonsmokers using flow cytofluorometry; Skold CM et al.; Flow cytofluorometry was used to compare blood monocytes (BMs) and alveolar macrophages (AMs) from the same nonsmoking subject (n = 13) . Autofluorescence was quantified, cell surface markers (HLA-DR, CR3) were detected by monoclonal antibodies, and phagocytic ability was determined using C3b-coated yeast particles . AMs expressed more HLA-DR (p less than 0.001) and CR3 (p less than 0.01) on their surfaces than did BMs . The phagocytic capacity was enhanced in AMs compared to BMs (p less than 0.001) and the cells showed an increased autofluorescence (p less than 0.001) in the alveoli compared to blood . The findings suggest that the mononuclear phagocyte is activated when it migrates from blood to alveoli in order to adapt to the milieu in the alveolar space. Mol Gen Genet, 1990 May, 221(3), 443 - 52 A Chlamydomonas gene encodes a G protein beta subunit-like polypeptide; Schloss JA; A Chlamydomonas gene encodes a protein that shows sequence similarity with the beta subunit of guanine nucleotide binding proteins from mammals, fruit fly and yeast . In addition to amino acid sequences similarity, each of these proteins contains a segmented repeat structure in which certain amino acids form a consensus sequence . Thus this gene product has been designated a Chlamydomonas beta subunit-like polypeptide (Cblp) . The mRNA is constitutively expressed during the cell cycle and during flagellar regeneration. Transfusion, 1990 May, 30(4), 374 - 6 Blood screening for non-A, non-B hepatitis by hepatitis C virus antibody assay; Katayama T et al.; Hepatitis C virus (HCV) antibody was detected in 1499 donor sera by radioimmunoassay using an antigen expressed in yeast from a cDNA clone of the HCV genome . Eighteen samples over 4200 counts per minute (cpm) were considered to contain infectious HCV because these recipients developed typical posttransfusion non-A, non-B hepatitis after transfusion . The antibody-positive sera were all within the normal range of ALT levels . This assay system is thus useful for the screening for blood transfusion. Proc Natl Acad Sci U S A, 1990 May, 87(10), 3821 - 5 Molecular cloning and characterization of GPA1, a G protein alpha subunit gene from Arabidopsis thaliana; Ma H et al.; We have isolated a gene coding for a G protein alpha subunit from the flowering plant Arabidopsis thaliana . This gene, named GPA1, was isolated by using a DNA probe generated by polymerase chain reaction based on protein sequences from mammalian and yeast G protein alpha subunits . The sequences of genomic and cDNA clones indicate that GPA1 has 14 exons, and the deduced amino acid sequence shows that the GPA1 gene product (GP alpha 1) has 383 amino acid residues (44,582 Da) . The GP alpha 1 protein exhibits similarity to all known G protein alpha subunits--36% of its amino acids are identical and 73% are similar (identical and conservative changes) to mammalian inhibitory guanine nucleotide-binding regulatory factor alpha subunits and transducins . Furthermore, the GP alpha 1 protein has all of the consensus regions for a GTP-binding protein . The GPA1-encoded mRNA of 1.55 kilobases is most abundant in vegetative plant tissues, as determined by RNA blot analysis . Restriction fragment length polymorphism mapping experiments show that GPA1 is approximately 1.2 centimorgans from the visible marker er on chromosome 2. J Cell Biol, 1990 May, 110(5), 1555 - 64 Cytosolic free calcium elevation mediates the phagosome-lysosome fusion during phagocytosis in human neutrophils; Jaconi ME et al.; Cytosolic free calcium ({Ca2+}i) and fusion of secondary granules with the phagosomal membrane (phagosome-lysosome fusion, P-L fusion) were assessed in single adherent human neutrophils during phagocytosis of C3bi-opsonized yeast particles . Neutrophils were loaded with the fluorescent dye fura2/AM and {Ca2+}i was assessed by dual excitation microfluorimetry . Discharge of lactoferrin, a secondary granule marker into the phagosome was verified by immunostaining using standard epifluorescence, confocal laser scanning and electron microscopy . In Ca2(+)-containing medium, upon contact with a yeast particle, a rapid rise in {Ca2+}i was observed, followed by one or more Ca2+ peaks (maximal value 1,586 nM and median duration 145 s): P-L fusion was detected in 80% of the cells after 5-10 min . In Ca2(+)-free medium the amplitude, frequency and duration of the {Ca2+}i transients were decreased (maximal value 368 nM, mostly one single Ca2+ peak and median duration 75 s): P-L fusion was decreased to 52% . Increasing the cytosolic Ca2+ buffering capacity by loading the cells with MAPT/AM led to a dose-dependent inhibition both of {Ca2+}i elevations and P-L fusion . Under conditions where basal {Ca2+}i was reduced to less than 20 nM and intracellular Ca2+ stores were depleted, P-L fusion was drastically inhibited while the cells ingested yeast particles normally . P-L fusion could be restored in Ca2(+)-buffered cells containing ingested particles by elevating {Ca2+}i with the Ca2(+)-ionophore ionomycin . The present findings directly indicate that although the ingestion step of phagocytosis is a Ca2(+)-independent event, {Ca2+}i transients triggered upon contact with opsonized particles are necessary to control the subsequent fusion of secondary granules with the phagosomal membrane. Zhongguo Yao Li Xue Bao, 1990 May, 11(3), 204 - 7 {Analgesic and antipyretic effects of cyproheptadine}; Tan JQ et al.; Cyproheptadine-HCl raised the pain thresholds during hot plat test and writhing test in mice and tail flick test in rats, strengthened the hypnotic action by subthreshold dosage of sodium pentobarbital and chloral hydrate . The ED50 were 4.4 (3.2-5.7) and 12.4 (8.4-18.2) mg/kg 30 min after ip cyproheptadine in mice and rats, respectively . The ED50 was 0.14 (0.12-0.18) mg/kg 90 min after icv cyproheptadine in mice . Cyproheptadine po 20, 40 mg/kg and ip 10, 20 mg/kg showed significant antipyretic effects on yeast-induced pyrexia in rats. Development, 1990 May, 109(1), 149 - 56 In vivo regulation of MPF in Xenopus oocytes; Johnson AD et al.; Entry into M phase in the eukaryotic cell cycle is controlled by the oscillating activity of MPF . The active component of MPF is now known to be the p34cdc2 protein kinase originally found in yeast . The p34cdc2 protein kinase displays a characteristic M-phase-specific histone H1 kinase activity when it interacts with cyclins, which are proteins that oscillate through the cell cycle and are thought to regulate p34cdc2 activity . Cyclins can induce M phase when introduced into fully grown Xenopus oocytes and cyclin may play a role in normal oocyte maturation . Small Xenopus oocytes do not mature in response to the hormonal triggers which act on stage 6 oocytes . We introduced cyclin into stage 4 (small) Xenopus oocytes and showed that it activates MPF in these cells, probably by interacting with endogenous p34cdc2 kinase . We made labelled extracts from cyclin-mRNA-injected stage 4 oocytes and used them to show differential stability of clam cyclins A and B at oocyte maturation . The relative stability of the two forms of cyclin related directly to their ability to stabilize crude MPF preparations from injected stage 6 oocytes. Trans R Soc Trop Med Hyg, 1990 May-Jun, 84(3), 425 - 8 Preparation of monoclonal antibodies that differentiate between Histoplasma capsulatum variant capsulatum and H . capsulatum variant duboisii; Hamilton AJ et al.; The immunosuppressive drug cyclophosphamide was used to facilitate the production of monoclonal antibodies (Mabs) which differentiated between the yeast phase of the two variants of the dimorphic fungus Histoplasma capsulatum by both enzyme-linked immunosorbent assay and Western blot . Two Mabs are described, identifying epitopes on a 70-75 kDa molecule, which are specific to H . capsulatum var . capsulatum and which do not identify epitopes of H . capsulatum var . duboisii . These Mabs have potential use in the epidemiology and serodiagnosis of histoplasmosis in areas where both classical and African forms of the disease occur. J Cell Sci, 1990 May, 96 ( Pt 1), 121 - 9 Monoclonal antibody analysis of the proliferating cell nuclear antigen (PCNA) . Structural conservation and the detection of a nucleolar form; Waseem NH et al.; The proliferating cell nuclear antigen, PCNA, has recently been identified as the polymerase delta accessory protein . PCNA is essential for cellular DNA synthesis and is also required for the in vitro replication of simian virus 40 (SV40) DNA where it acts to coordinate leading and lagging strand synthesis at the replication fork . The cDNA for rat PCNA was cloned into a series of bacterial expression vectors and the resulting protein used to immunize mice . Eleven new monoclonal antibodies to PCNA have been isolated and characterized . Some of the antibodies recognize epitopes conserved from man to fission yeast . Immunocytochemical analysis of primate epithelial cell lines showed that the antibodies recognized antigenically distinct forms of PCNA and that these forms were localized to different compartments of the nucleus . One antibody reacted exclusively with PCNA in the nucleolus . These results suggest that the PCNA protein may fulfil several separate roles in the cell nucleus associated with changes in its antigenic structure. J Am Acad Dermatol, 1990 May, 22(5 Pt 1), 739 - 42 Immune reactions to Pityrosporum ovale in adult patients with atopic and seborrheic dermatitis; Kieffer M et al.; Pityrosporum ovale is a lipophilic yeast commonly present in the seborrheic areas of the skin of adults . Fifty-five young adult patients with atopic dermatitis, 19 patients with seborrheic dermatitis, and 19 healthy control subjects were examined for immune reactions to P . ovale, including tests for specific IgE antibodies (prick test, histamine release), IgG antibodies and epicutaneous testing . IgE antibodies against P . ovale were found in two thirds of the patients with atopic dermatitis and were more frequent in patients with lesions predominantly in the seborrheic areas . In addition, some atopic patients had positive reactions to epicutaneous tests, which suggest that delayed allergic reactions to P . ovale may also be important . In patients with seborrheic dermatitis, no evidence of immediate or delayed hypersensitivity to P . ovale was found . IgG antibody levels were low in all groups. Eur J Biochem, 1990 Apr 30, 189(2), 259 - 65 The conformation of single-stranded nucleic acids tDNA versus tRNA; Paquette J et al.; Conformational analyses using the single-strand-specific nuclease from mung bean and restriction endonucleases have been performed on a series of DNA fragments related to the sequence of the yeast initiator tRNA(Met) . Mung bean nuclease cleaves DNA fragments exclusively in some, but not all, single-stranded regions as predicted by RNA secondary structural rules . Comparison of cleavage patterns of yeast initiator tRNA(Met), tDNA(Met) (a DNA oligomer having the sequence of tRNA(Met} and the anti-tDNA(Met) (the complement of tDNA(Met} suggests that the conformation of the three molecules is very similar . Furthermore, both tDNA and anti-tDNA are cleaved by HhaI and CfoI restriction endonucleases at two GCG/C sites which would be in double-stranded regions (the acceptor and dihydrouridine stem), if the two molecules adopt the tRNA cloverleaf structure . On the other hand, minor cleavage products show that the core region, i.e . the extra loop area, is slightly more exposed in tDNA and in anti-tDNA than in tRNA . Therefore, we submit that the global conformation of nucleic acids is primarily dictated by the interaction of purine and pyrimidine bases with atoms and functional groups common to both RNA and DNA . In this view the 2'-hydroxyl group, in tRNA at least, is an auxiliary structural feature whose role is limited to fostering local interactions, which increase the stability of a given conformation. Biochemistry, 1990 Apr 24, 29(16), 3966 - 72 The formation of covalent adducts between benzo{a}pyrenediol epoxide and RNA: structural analysis by mass spectrometry; Yamamoto J et al.; Racemic 7-r,8-t-dihydroxy-9-t,10-t-epoxy-7,8,9,10-tetrahydrobenzo{a} pyrene was reacted with yeast RNA . Modified nucleosides were isolated and resolved by high-performance liquid chromatography; nine adduct peaks were collected for analysis . The bases in these adducts were identified by comparing their retention times with those of adducts from poly(G), poly(A), and poly(C) . These samples gave two major and two minor Guo adducts, four major Ado adducts, and at least four Cyd adducts . The relative efficiencies of adduct formation with the polyribonucleotides were poly(G) greater than yeast RNA greater than poly(A) greater than poly(C) . Fluorescence measurements show that emission from Guo adducts is strongly quenched relative to that from Ado adducts . Liquid secondary ion mass spectrometry (LSIMS) of underivatized samples and electron-impact mass spectrometry (EIMS) of permethyl derivatives were used to confirm the base identities and establish the alkylation sites of the RNA adducts . Unique nitrogen-containing hydrocarbon fragments that were observed with all samples by EIMS establish that in each adduct analyzed the C-10 position of the hydrocarbon is linked to the exocyclic amino group of the base . This suggested that the multiple adducts formed with each base are diastereomers derived from cis/trans epoxide ring opening of the (+) and (-) enantiomers of the carcinogen . Several adducts exhibited molecular ions by both LSIMS and EIMS . Large fragments observed by EIMS usually resulted from the loss of CH3OH, CH3O., CH2O, CH3., and H . from the molecular ion . Major fragmentation pathways also resulted in formation of nucleoside, base, ribose, hydrocarbon, and base-hydrocarbon ions . Each of these major ions in turn resulted in further characteristic fragmentation patterns. Biochem Biophys Res Commun, 1990 Apr 16, 168(1), 22 - 9 Deficient lysosomal carboxypeptidase activity in galactosialidosis; Tranchemontagne J et al.; In the lysosome, the glycosidases neuraminidase (EC 3.2.1.18) and beta-galactosidase (EC 3.2.1.23) are associated to a 52 kDa "protective protein" to form a large multi-enzymatic complex . Deficient synthesis or inactivation of this protective protein causes galactosialidosis, a lysosomal storage disorder in man in which both neuraminidase and beta-galactosidase activities are deficient . Since the protective protein possesses extensive sequence homology with carboxypeptidase Y (carb Y) and the KEX 1 gene product from yeast, we have used the artificial substrate N-CBZ-Phe-Leu to detect and characterize the peptidase activity of the lysosomal carboxypeptidase (carb L) . Using both a purified preparation of the lysosomal multi-enzymatic complex and cultured skin fibroblasts of patients affected with galactosialidosis, we demonstrate that the 52 kDa protective protein is responsible for carb L activity . The fibroblasts of three patients affected with late infantile and juvenile galactosialidosis were found to be deficient in carb L activity (1.4% of normal mean value). J Biol Chem, 1990 Apr 15, 265(11), 6369 - 75 Selective and tandem amplification of a member of the metallothionein gene family in Candida glabrata; Mehra RK et al.; Metallothioneins constitute a multigene family in the yeast Candida glabrata . Two genes, designated metallothionein-I (MT-I) and one member of the metallothionein-II family (MT-II), were cloned and sequenced previously (Mehra, R . K., Garey, J . R., Butt, T . R., Gray, W . R., and Winge, D . R . (1989) J . Biol . Chem . 264, 19747-19753) . Southern analysis of the genomic DNA samples from different wild-type isolates indicated that the MT-I gene was always present as a single copy but multiple (3-9) and tandemly arranged copies of one MT-II gene were present in different strains . Strains of C . glabrata highly resistant to copper salts were obtained by repeated culturing of wild-type isolates in medium containing increasing concentrations of copper sulfate . These strains showed further stable chromosomal amplification (greater than 30 copies) of the MT-II gene . The MT-I gene remained as a single copy . Amplified copies of the MT-II gene were always arranged tandemly . One of the copper-resistant strains acquired more copies of the MT-II gene by apparent duplication of the chromosome carrying this gene . The size of the amplification unit was 1.25 kilobases . The principal MT-I and -II genes of C . glabrata were shown to map to different chromosomes by electrophoretic karyotypic analysis . The length of chromosome carrying MT-II gene increased appreciably in strains exhibiting the highest amplification of this gene . Northern analysis showed increased basal levels of MT-II mRNA in strains having highly amplified MT-II locus. Nature, 1990 Apr 5, 344(6266), 556 - 9 Sequencing and expression of complementary DNA for the general transcription factor BTF3; Zheng XM et al.; The initiation of transcription of eukaryotic genes involves the ordered assembly of a multiprotein complex on proximal promoter elements such as the TATA box . In addition to RNA polymerase II (otherwise RNA pol II, RNA polymerase B), four general transcription factors are required for initiation of transcription: BTF1 (also referred to as TFIID) which has recently been cloned from yeast, BTF2, BTF3 and STF . The first step in assembly of the initiation complex is the stable binding of BTF1 to the TATA box, which is facilitated by STF . Neither BTF2 nor BTF3 bind directly to the promoter proximal elements, but BTF3 can form a stable complex with RNA pol II . We recently purified BTF3, which is a protein of relative molecular mass 27,000, but further studies have been hampered by its low abundance in cells . On the basis of sequences from peptides of BTF3, we have now cloned two complementary DNAs, one for a protein (BTF3a) with all the characteristics of purified BTF3, and one for a shorter protein (BTF3b) lacking the first 44 residues of BTF3a and which is transcriptionally inactive, despite its ability to bind RNA pol II. Biochemistry, 1990 Apr 3, 29(13), 3198 - 202 Orotidine-5'-monophosphate decarboxylase catalysis: kinetic isotope effects and the state of hybridization of a bound transition-state analogue; Acheson SA et al.; The enzymatic decarboxylation of orotidine 5'-monophosphate may proceed by an addition-elimination mechanism involving a covalently bound intermediate or by elimination of CO2 to generate a nitrogen ylide . In an attempt to distinguish between these two alternatives, 1-(phosphoribosyl)barbituric acid was synthesized with 13C at the 5-position . Interaction of this potential transition-state analogue inhibitor with yeast orotidine-5'-monophosphate decarboxylase resulted in a small (0.6 ppm) downfield displacement of the C-5 resonance, indicating no rehybridization of the kind that might have been expected to accompany 5,6-addition of an enzyme nucleophile . When the substrate orotidine 5'-monophosphate was synthesized with deuterium at C-5, no significant change in kcat (H/D = 0.99 +/- 0.06) or kcat/KM (H/D = 1.00 +/- 0.06) was found to result, suggesting that C-5 does not undergo significant changes in geometry before or during the step that determines the rate of the catalytic process . These results are consistent with a nitrogen ylide mechanism and offer no support for the intervention of covalently bound intermediates in the catalytic process. Sheng Li Xue Bao, 1990 Apr, 42(2), 163 - 8 {The inhibitory effect of hydrocortisone on the chemotactic migration of human leukocytes}; Liu ZM et al.; Random migration (RM) and chemotactic migration (ChtM) of human leukocytes to yeast activated serum were studied with the modified Boyden chamber method . Both RM and ChtM showed circadian rhythm . Leukocytes migrated most rapidly at night . The difference between the peak (0:00) and trough values (8:00) of RM and ChtM was significant statistically (P less than 0.01) . ChtM was inhibited by hydrocortisone (F) of physiological concentration (10(-9)-10(-7) mol/L) which was dose-dependent and completely antagonized by the competitive antagonist, RU38486 . The inhibitory effect was much more evident with higher dose (more than 10(-5) mol/L) and it was also reversed by RU38486, but only partially . It is suggested that glucocorticoids (GC) may be a physiological regulator of the activity of leukocytes and its inhibitory action on ChtM may be involved in antiinflammatory mechanisms of GC of pharmacological doses . The action of physiological and pharmacological concentration of GC may be mediated by low affinity specific binding sites of glucocorticoid receptors. J Biochem (Tokyo), 1990 Apr, 107(4), 546 - 9 Differentiation of human non-salivary, non-pancreatic alpha-amylase from salivary and pancreatic alpha-amylases by use of FG5P; Omichi K et al.; Human non-salivary, non-pancreatic alpha-amylase (yHXA) is the gene product of a newly found human alpha-amylase gene expressed in yeast . Its mode of action on a fluorogenic derivative of p-nitrophenyl alpha-maltopentaoside, FG5P (FG-G-G-G-G-P), was examined at various pH values to elucidate the difference between yHXA and pancreatic or salivary alpha-amylase . The product analysis of the digests by HPLC showed that the enzyme hydrolyzed FG5P to FG3 (FG-G-G) and p-nitrophenyl alpha-maltoside (G-G-P) and to FG4(FG-G-G-G) and p-nitrophenyl alpha-glucoside (G-P), and the ratio of the two reactions changed with pH . The three enzymes differed from each other in the mode of action at pH 5.5 . The molar ratio of FG4 to FG3 in the digest with yHXA was the largest . This suggested that the expression of the new gene in human can be detected by the use of FG5P as the substrate in the alpha-amylase assay. Arch Biochem Biophys, 1990 Apr, 278(1), 65 - 72 Extramitochondrial release of hydrogen peroxide from insect and mouse liver mitochondria using the respiratory inhibitors phosphine, myxothiazol, and antimycin and spectral analysis of inhibited cytochromes; Bolter CJ et al.; The fumigant insecticide phosphine (PH3) is known to inhibit cytochrome c oxidase in vitro . Inhibition of the respiratory chain at this site has been shown to stimulate the generation of superoxide radicals (O2-), which dismutate to form hydrogen peroxide (H2O2) . This study was performed in order to investigate the production of H2O2 by mitochondria isolated from granary weevil (Sitophilus granarius) and mouse liver on exposure to PH3 . Other respiratory inhibitors, antimycin, myxothiazol, and rotenone were used with insect mitochondria . Hydrogen peroxide was measured spectrophotometrically using yeast cytochrome c peroxidase as an indicator . Insect and mouse liver mitochondria, utilizing endogenous substrate, both produced H2O2 after inhibition by PH3 . Insect organelles released threefold more H2O2 than did mouse organelles, when exposed to PH3 . Production of H2O2 by PH3-treated insect mitochondria was increased significantly on addition of the substrate alpha-glycerophosphate . Succinate did not enhance H2O2 production, however, indicating that the H2O2 did not result from the autoxidation of ubiquinone . NAD(+)-linked substrates, malate and pyruvate also had no effect on H2O2 production, suggesting that NADH-dehydrogenase was not the source of H2O2 . Data obtained using antimycin and myxothiazol, both of which stimulated the release of H2O2 from insect mitochondria, lead to the conclusion that glycerophosphate dehydrogenase is a source of H2O2 . The effect of combining PH3, antimycin, and myxothiazol on cytochrome spectra in insect mitochondria was also recorded . It was observed that PH3 reduces cytochrome c oxidase but none of the other cytochromes in the electron transport chain . There was no movement of electrons to cytochrome b when insect mitochondria are inhibited with PH3 . The spectral data show that the inhibitors interact with the respiratory chain in a way that would allow the production of H2O2 from the sites proposed previously. Diabetes, 1990 Apr, 39(4), 432 - 6 Effect of diet on incidence of diabetes in nonobese diabetic mice; Coleman DL et al.; Nonobese diabetic/Lt mice exhibit a diabetes incidence greater than 70% in females at 30 wk of age . In studies designed to see whether increased dietary carbohydrate, fat, or protein influenced the severity or age at onset of the syndrome, we fed semipurified AIN-76 diet adulterated with increased amounts of these ingredients . Surprisingly, all AIN-76-based diets greatly reduced the expected incidence of diabetes at 30 wk . In addition, a hypoallergenic infant formula, Pregestimil, containing casein hydrolysate in place of protein, completely prevented diabetes up to 1 yr of age . To assess how dietary components might modulate the diabetes incidence, we adulterated standard AIN-76 diet with skim milk, gluten, brewer's yeast, or a natural-ingredient rodent open-formula mouse diet (Old Guilford 96 {OG96} . No increase in diabetes incidence was seen with skim milk (10%) or wheat gluten (10%), whereas brewer's yeast (10%) and OG96 (25%) added to AIN-76 increased the incidence compared to mice fed OG96 only . The diabetogenic factor or factors in OG96 could be extracted by chloroform plus methanol (2:1), leaving little activity in the residue . We conclude that diet is a critical factor in diabetes development and that unknown chloroform-methanol-soluble substances in natural-ingredient chow not found in semipurified diets can enhance the development of diabetes in genetically susceptible mice. J Cell Sci, 1990 Apr, 95 ( Pt 4), 521 - 6 Telomere cloning and mammalian chromosome analysis; Brown WR et al.; Although eucaryotic chromosomes vary in size over five orders of magnitude and are constituents of diverse genetic systems the fundamental features of their telomeres appear to be almost completely conserved . This can be exploited to enable molecular cloning of human telomeres in yeast and suggests that many of the ideas that will arise from studies of telomeres in the experimentally tractable ciliates and yeasts will hold true of mammalian telomeres . The particular value of cloned mammalian telomeres is that they contribute reagents for mapping mammalian chromosomes and that they provide one set of elements for the construction of artificial mammalian chromosomes. Trends Biochem Sci, 1990 Apr, 15(4), 139 - 42 Prenyl proteins in eukaryotic cells: a new type of membrane anchor; Glomset JA et al.; Recent studies have indicated that eukaryotic cells contain proteins that are post-translationally modified by long-chain, thioether-linked prenyl groups . These proteins include yeast mating factors, ras proteins and nuclear lamins . The modification occurs on a cysteine residue near the C terminus and appears to initiate a set of additional protein modification reactions that promote attachment of the proteins to specific membranes. Antonie Van Leeuwenhoek, 1990 Apr, 57(3), 179 - 89 Chemotaxonomy of the genus Talaromyces; Frisvad JC et al.; Species of the ascomycetous genus Talaromyces have been examined for profiles of secondary metabolites on TLC . The greatest number of specific metabolites were produced on oatmeal-, malt extract- and yeast-extract sucrose agars . Profiles of intracellular secondary metabolites produced on oatmeal agar were specific for each species and provided a means of simple differentiation of the taxa . Examination of the most important species using high performence liquid chromatography (HPLC) allowed to solve some taxonomic problems . Known mycotoxins are produced by T . stipitatus (duclauxin, talaromycins, botryodiploidin), T . stipitatus chemotype II (emodin), T . panasenkoi (spiculisporic acid), T . trachyspermus (spiculisporic acid), T . macrosporus (duclauxin) and T . wortmannii (rugulosin) . Wortmannin is produced by an atypical strain of T . flavus but not T . wortmannii . Several other secondary metabolites were discovered for the first time in the following species: Glauconic acid is produced by T . panasenkoi, T . ohiensis and T . trachyspermus; vermiculine by T . ohiensis; duclauxin by T . flavus var . macrosporus and the mitorubrins by T . flavus and T . udagawae . The profiles of secondary metabolites support the established taxonomy of the species based on morphology, showing the genetic stability of profiles of secondary metabolites in Talaromyces . Two new taxa are proposed: T . macrosporus comb . nov . (stat . anam . Penicillium macrosporum stat . nov.), and Penicillium vonarxii, sp . nov . for the anamorph of T . luteus. Obstet Gynecol Surv, 1990 Apr, 45(4), 220 - 8 The premenstrual syndrome; Lurie S et al.; PMS is probably a group of entities which include various symptoms that occur during the 7 to 10 days before menstruation and disappear a few hours after the onset of menstruation . The definition of PMS lacks objective criteria . The most common symptoms are irritability, bloating, aggressiveness, mastodynia, and headaches . The prevalence of PMS is estimated at 30 to 40 per cent . PMS is more prevalent among women working outside the home, alcoholics, women of high parity, and women with toxemic tendency; it probably runs in families . The etiology of PMS is no less obscure to us than when it was first described by Frank in 1931 . No single theory has been established to explain the entire diversity of PMS symptomatology . The multitude of possible etiologic factors includes psychosocial bases, progesterone deficiency, prolactin excess, thyroid hypofunction, renin angiotensin alternations, antidiuretic hormone excess, decreased colloidosmotic pressure, endorphin activity alternations, serotonin metabolism alternations, prostaglandin action, vitamin deficiency, and such unconventional theories as the ovarian infection or the "yeast overgrowth" theory . A partial resolution of this divergence of hypotheses comes from the biopsychosocial model developed by Keye and Trunnel . According to this model, a biologic, perhaps genetically determined, predisposition to PMS is realized when past and present life experiences, attitudes, beliefs, coping styles, and social forces interact to stress a woman . The diagnosis of PMS is based on establishing a relationship between the luteal phase of the cycle and the symptoms . The evaluation of PMS patients includes the use of a monthly diary to scale the symptoms, a physical examination, and biochemical studies to rule out other disorders . Management includes education, reassurance, and drug therapy.(ABSTRACT TRUNCATED AT 250 WORDS) Jpn J Med Sci Biol, 1990 Apr, 43(2), 29 - 36 Prevalence of hepatitis C virus antibody among healthy blood donors and non-A, non-B hepatitis patients in Thailand; Boonmar S et al.; With the antigen expressed in yeast from a cDNA clone encoding a non-structural region of newly discovered hepatitic C virus (HCV) genome, the prevalence of HCV antibody in people in Thailand was investigated . Antibody was detected in 2.6% of healthy blood donors and in 2.8% of healthy pregnant women . These prevalence rates were higher than those reported previously from Japan, USA and European countries . Among community-acquired, sporadic cases of acute and chronic non-A, non-B hepatitis, however, only 5.7% and 15.4% were shown to possess the antibody, respectively . Among hepatocellular carcinoma patients who were negative for hepatitis B surface antigen in the sera, 11.1% had antibody to HCV . These seroepidemiological data suggest that HCV plays an important role as an etiological agent in Thailand; however, other agents must also be involved in etiologic agents of viral hepatitis and chronic liver disease. Development, 1990 Apr, 108(4), 525 - 42 Calcium and cell cycle control; Whitaker M et al.; The cell division cycle of the early sea urchin embryo is basic . Nonetheless, it has control points in common with the yeast and mammalian cell cycles, at START, mitosis ENTRY and mitosis EXIT . Progression through each control point in sea urchins is triggered by transient increases in intracellular free calcium . The Cai transients control cell cycle progression by translational and post-translational regulation of the cell cycle control proteins pp34 and cyclin . The START Cai transient leads to phosphorylation of pp34 and cyclin synthesis . The mitosis ENTRY Cai transient triggers cyclin phosphorylation . The motosis EXIT transient causes destruction of phosphorylated cyclin . We compare cell cycle regulation by calcium in sea urchin embryos to cell cycle regulation in other eggs and oocytes and in mammalian cells. Mol Gen Genet, 1990 Apr, 221(2), 148 - 54 Nucleotide sequence and nuclear protein binding of the two regulatory sequences upstream of the am (GDH) gene in Neurospora; Frederick GD et al.; We have constructed a series of deletions in the 5' non-coding sequences of the cloned Neurospora crassa am gene which specifies NADP specific glutamate dehydrogenase . All of the deletions begin at -4.4 kb with respect to the am transcription start site and extend for various distances toward the am gene . Using vectors with a truncated fragment of the am gene, we introduced these deletions into the chromosome upstream of am by transformation . Analysis of glutamate dehydrogenase expression in strains with the deletion mutations confirmed that there are two upstream regulatory sequences (URS) that control the expression of the am gene . The more distal of these elements (URSam beta) has been limited to the 157 bp between -1924 and -2081 with respect to the start of am transcription . The proximal element (URSam alpha) was limited to the 97 bp between -1296 and -1393 . The DNA sequence of the entire region was determined . Within the sequences that contain the URS elements several regions of homology with yeast UAS sequences were found . Gel mobility assays with DNA fragments containing the URS elements indicated that sequences in both elements are bound by nuclear proteins from Neurospora . The interaction of these proteins and the DNA fragments was found to be specific. Biochem Biophys Res Commun, 1990 Mar 30, 167(3), 1214 - 20 Regulation of phosphofructokinase at physiological concentration of enzyme studied by stopped-flow measurements; Bar J et al.; Stopped-flow measurements have been carried out to study some basic allosteric properties of muscle and yeast phosphofructokinase at physiological concentration of enzyme . An important increase in the affinity for fructose-6-P accompanied by an intense decrease in the ATP inhibition was observed with the muscle enzyme, which also became insensitive to fructose-2,6-P2 under these conditions . Yeast phosphofructokinase exhibited a significant diminution in the inhibition by ATP, although with no apparent change in the affinity for fructose-6-P . These results provide strong support in favor of the dependence of the allosteric regulation of phosphofructokinase on its concentration in vivo. Mol Cell Biochem, 1990 Mar 27, 93(2), 153 - 65 Phosphoglucoisomerase-catalyzed interconversion of hexose phosphates: isotopic discrimination between hydrogen and deuterium; Malaisse WJ et al.; The discrimination between the isotopes of hydrogen in the reaction catalyzed by yeast phosphoglucoisomerase is examined by NMR, as well as by spectrofluorometric or radioisotopic methods . The monodirectional conversion of D-glucose 6-phosphate to D-fructose 6-phosphate displays a lower maximal velocity with D-{2-2H}glucose 6-phosphate than unlabelled D-glucose 6-phosphate, with little difference in the affinity of the enzyme for these two substrates . About 72% of the deuterium located on the C2 of D-{1-13C,2-2H}glucose 6-phosphate is transferred intramolecularly to the C1 of D-{1-13C,1-2H}fructose 6-phosphate . The velocity of the monodirectional conversion of D-{U-14C}glucose 6-phosphate (or D-{2-3H}glucose 6-phosphate) to D-fructose 6-phosphate is virtually identical in H2O and D2O, respectively, but is four times lower with the tritiated than 14C-labelled ester . In the monodirectional reaction, the intramolecular transfer from the C2 of D-{2-3H}glucose 6-phosphate is higher in the presence of D2O than H2O . Whereas prolonged exposure of D-{1-13C}glucose 6-phosphate to D2O, in the presence of phosphoglucoisomerase, leads to the formation of both D-{1-13C,2-2H}glucose 6-phosphate and D-{1-13C,1-2H}fructose 6-phosphate, no sizeable incorporation of dueterium from D2O on the C1 of D-{1-13C}fructose 1,6-bisphosphate is observed when the monodirectional conversion of D-{1-13C}glucose 6-phosphate occurs in the concomitant presence of phosphoglucoisomerase and phosphofructokinase . The latter finding contrasts with the incorporation of hydrogen from 1H2O or tritium from 3H2O in the monodirectional conversion of D-{2-3H}glucose 6-phosphate and unlabelled D-glucose 6-phosphate, respectively, to their corresponding ketohexose esters. Nucleic Acids Res, 1990 Mar 25, 18(6), 1451 - 5 Human ureaplasmas show diverse genome sizes by pulsed-field electrophoresis; Robertson JA et al.; Contour clamped homogeneous field (CHEF) agarose gel electrophoresis (AGE), ramped to give linear separation of DNA molecules of 600-1600 kilobase pairs (kbp), was used to determine mobilities for full-sized genomic DNA of the serotype standard strains of the human genital mollicutes, Ureaplasma urealyticum relative to yeast chromosomal DNA markers . Indicated genome sizes (in kbp) were 760 for the four biotype 1 strains and 840-1140 for eleven biotype 2 strains . Other estimates were: 720 for Mycoplasma hominis, 1070 for Mycoplasma hyopneumoniae, 890 for Mycoplasma flocculare, 1180 and 1350 for Mycoplasma mycoides subsp . mycoides Y and GC1176-2, respectively, and 1650 and 1580 for Acholeplasma laidlawii B and PG 8, respectively . These data supplement previous evidence from CHEF AGE that the genomes of the Mycoplasmataceae are diverse in size with some larger than previously estimated from DNA renaturation kinetics. Cell, 1990 Mar 23, 60(6), 991 - 7 The candidate proto-oncogene bcl-3 is related to genes implicated in cell lineage determination and cell cycle control; Ohno H et al.; A gene, bcl-3, is found on chromosome 19 adjacent to the breakpoints in the translocation t(14;19)(q32;q13.1), which occurs in some cases of chronic lymphocytic leukemia . Sequence analysis of the human bcl-3 gene predicts a protein containing seven tandem copies of the SWI6/cdc10 motif . This motif was previously identified in yeast genes that regulate events at the start of the cell cycle and in invertebrate transmembrane proteins involved in cell differentiation pathways . Expression of bcl-3 in normal blood cells increases markedly following mitogenic stimulation, and leukemic cells with the translocation show much greater expression than controls . These results suggest that bcl-3 is a proto-oncogene that may contribute to leukemogenesis when abnormally expressed. J Mol Biol, 1990 Mar 20, 212(2), 259 - 68 Identification of a Caenorhabditis elegans histone H1 gene family . Characterization of a family member containing an intron and encoding a poly(A)+ mRNA; Sanicola M et al.; The isolation and properties of a gene encoding a histone H1 protein of Caenorhabditis elegans, his-24, are described . The predicted protein sequence is similar to histone H1 proteins of other eukaryotes . However, the gene structure of his-24 is atypical for a histone H1 gene; it contains an intron and encodes a polyadenylated mRNA . A family of approximately five histone H1 genes is defined by cross-hybridization to his-24 . All appear to encode polyadenylated mRNAs . One gene is expressed specifically in male germ cells . These histone H1 genes are dispersed individually in the genome, apart from the previously described clusters of core histone genes (H2A, H2B, H3 and H4), which probably all encode non-polyadenylated mRNAs . This histone gene organization, with clustered core histone genes, encoding non-polyadenylated transcripts, and dispersed, histone H1 genes from which it appears only polyadenylated messages arise, suggests that C . elegans is at a stage of evolution of the histone gene family intermediate between lower eukaryotes (e.g . yeast) and the most advanced forms. Cancer Res, 1990 Mar 15, 50(6), 1905 - 10 Effects of microinjected photoreactivating enzyme on thymine dimer removal and DNA repair synthesis in normal human and xeroderma pigmentosum fibroblasts; Roza L et al.; UV-induced thymine dimers (10 J/m2 of UV-C) were assayed in normal human and xeroderma pigmentosum (XP) fibroblasts with a monoclonal antibody against these dimers and quantitative fluorescence microscopy . In repair-proficient cells dimer-specific immunofluorescence gradually decreased with time, reaching about 25% of the initial fluorescence after 27 h . Rapid disappearance of dimers was observed in cells which had been microinjected with yeast photoreactivating enzyme prior to UV irradiation . This photoreactivation (PHR) was light dependent and (virtually) complete within 15 min of PHR illumination . In general, PHR of dimers strongly reduces UV-induced unscheduled DNA synthesis (UDS) . However, when PHR was applied immediately after UV irradiation, UDS remained unchanged initially; the decrease set in only after 30 min . When PHR was performed 2 h after UV exposure, UDS dropped without delay . An explanation for this difference is preferential removal of some type(s) of nondimer lesions, e.g., (6-4) photoproducts, which is responsible for the PHR-resistant UDS immediately following UV irradiation . After the rapid removal of these photoproducts, the bulk of UDS is due to dimer repair . From the rapid effect of dimer removal by PHR on UDS it can be deduced that the excision of dimers up to the repair synthesis step takes considerably less than 30 min . Also in XP fibroblasts of various complementation groups the effect of PHR was investigated . The immunochemical dimer assay showed rapid PHR-dependent removal comparable to that in normal cells . However, the decrease of (residual) UDS due to PHR was absent (in XP-D) or much delayed (in XP-A and -E) compared to normal cells . This supports the idea that in these XP cells preferential repair of nondimer lesions does occur, but at a much lower rate. J Biol Chem, 1990 Mar 15, 265(8), 4678 - 84 Mouse "protective protein" . cDNA cloning, sequence comparison, and expression; Galjart NJ et al.; The "protective protein" is the glycoprotein that forms a complex with the lysosomal enzymes beta-galactosidase and neuraminidase . Its deficiency in man leads to the metabolic storage disorder galactosialidosis . The primary structure of human protective protein, deduced from its cloned cDNA, shows homology to yeast serine carboxypeptidases . We have isolated a full-length cDNA encoding murine protective protein . The nucleotide sequences as well as the predicted amino acid sequences are highly conserved between man and mouse . Domains important for the protease function are completely identical in the two proteins . Both human and mouse mature protective proteins covalently bind radiolabeled diisopropyl fluorophosphate . Transient expression of the murine cDNA in COS-1 cells yields a protective protein precursor of 54 kDa, a size characteristic of the glycosylated form . This cDNA-encoded precursor, endocytosed by human galactosialidosis fibroblasts, is processed into a 32- and a 20-kDa heterodimer and corrects beta-galactosidase and neuraminidase activities . A tissue-specific expression of protective protein mRNA is observed when total RNA from different mouse organs is analyzed on Northern blots. Med Clin (Barc), 1990 Mar 10, 94(9), 329 - 32 {Evaluation of an agglutination method of latex particles sensitized for the detection of vaginal candidiasis}; Cabronero MC et al.; We assayed clinically and experimentally a rapid method for detection of Candida in vagina by slide latex agglutination test (SLA) . A total of 50 vaginal swabs from women with clinical evidence of candidiasis were examined for yeast by microscopical examination, mycological culture and compared with the Candida antigen test . We also studied experimentally the sensibility of the SLA test . We found better results with SLA than microscopy for rapid diagnosis . Of the women with certain vaginal candidiasis, 60% were diagnosed by microscopy and 100% by SLA . SLA provided similar information than culture, but is faster and the clinician can use its information choosing the appropriate antifungal treatment. J Mol Biol, 1990 Mar 5, 212(1), 113 - 25 Effect of deletions at structural domains of group II intron bI1 on self-splicing in vitro; Bachl J et al.; Some group II introns can undergo a protein-independent splicing reaction with the basic reaction pathway similar to nuclear pre-mRNA splicing and the catalytic functions of some of the structural components have been determined . To identify further functional domains, we have generated an ensemble of partial and complete deletions of domains I, II, III and IV of the self-splicing group II intron bI1 from yeast mitochondria and studied their effects on the splicing reaction in vitro . Our results indicate that domains II and IV, which vary considerably in length and structure among group II introns, do not play a direct role in catalysis but mainly help to ensure the proper interaction between upstream and downstream catalytically active structural elements . Deletions of sub-domains of domain I and domain III indicate that these elements are involved in 5' cleavage by hydrolysis and in a reaction in trans (exon reopening), and that this function can be inhibited without affecting the normal 5' cleavage by transesterification . Yet, we infer that the helical structures affected by the mutational alterations might not contribute to this reaction mode per se but that changes within local secondary structures perturb the internal conformation of the ribozyme . Furthermore, we have designed an abbreviated version of intron bI1, with a length of 542 nucleotides, which is still catalytically active. Physiol Behav, 1990 Mar, 47(3), 409 - 13 Olfactory self-selection of protein-containing foods; Heinrichs SC et al.; When one unflavored, nonprotein diet was available in two differently scented bins, rats fed a protein-free diet over four days ate more from the bin smelling of gluten, ovalbumin, yeast or fibrin, but not soy, casein or lactalbumin, than from the bin smelling of butter . Rats fed a protein-containing diet over the same four-day period had no such preference . This result demonstrates that protein-deprived rats can use odor cues in making their selection of certain proteins . Since the direction, speed, and size of preference for these protein odors, excepting soy, are remarkably similar to those previously observed when rats actually consumed the proteins, olfactory stimuli appear to elicit appropriate protein selection responses independently of other protein quality variables such as taste, texture or nutrient composition. J Ethnopharmacol, 1990 Mar, 28(3), 305 - 12 Studies on Ruta chalepensis, an ancient medicinal herb still used in traditional medicine; al-Said MS et al.; An ethanolic extract of the aerial parts of Ruta chalepensis was studied for its anti-inflammatory, antipyretic, analgesic and CNS depressant activities . The extract produced a significant inhibition of carrageenan-induced paw oedema and cotton pellet granuloma in rats . The studies on spontaneous motor activity in mice and conditioned avoidance responding (CAR) in rats showed a dose-dependent depression of the central nervous system in treated animals . Reduction of yeast-induced hyperthermia in mice confirmed its reputed antipyretic activity . The extract did not produce any significant changes in prothrombin time and fibrinogen level . It also failed to produce any analgesic activity in the hot plate reaction-time test in mice . Phytochemical screening of the aerial parts of the plant showed the presence of alkaloids, flavonoids, coumarins, tannins, volatile oil, sterols and/or triterpenes. Methods Find Exp Clin Pharmacol, 1990 Mar, 12(2), 141 - 8 Pharmaco-toxicological effects of acetaminophen in rodents . Battery of tests to screen potential analgesic acetaminophen derivatives; Guasch J et al.; The pharmacological effects of acute oral administration of acetaminophen have been extensively evaluated in rodents . Using this drug as reference compound we have also standardized several pharmaco-toxociological tests in order to select new analgesic-antipyretic acetaminophen derivatives . According to the results obtained in this work, the following battery of assays is proposed to screen these compounds: a) pharmacological: antipyretic activity against Brewer's yeast-induced pyresis in rats; analgesic activity against chemical-induced writhings in mice and paw pressure test in rats; b) toxicological: oral acute toxicity in mice with and without phenobarbital pretreatment; determination of SGOT and SGPT activities as well as total bilirubin in mice . The usefulness of this procedure in the evaluation of drug effects is also considered. Clin Pharmacol Ther, 1990 Mar, 47(3), 313 - 22 Polymorphism in hydroxylation of mephenytoin and hexobarbital stereoisomers in relation to hepatic P-450 human-2; Yasumori T et al.; Stereoselective 4'-hydroxylations of R-(-)-mephenytoin and S-(+)-mephenytoin and 3'-hydroxylation of R-(-)-hexobarbital and S-(+)-hexobarbital were determined in liver microsomes of 14 Japanese subjects who were extensive metabolizers of mephenytoin and in five Japanese subjects who were poor metabolizers of mephenytoin . Content of P-450 human-2 assessed by Western blots was correlated to microsomal S-(+)-mephenytoin 4'-hydroxylation, R-(-)-hexobarbital 3' alpha-hydroxylation, and S-(+)-hexobarbital 3' beta-hydroxylation, and was less correlated to R-(-)mephenytoin 4'-hydroxylation, R-(-)-hexobarbital 3' beta-hydroxylation, and S-(+)-hexobarbital 3' alpha-hydroxylation . Antibodies raised against P-450 human-2 inhibited microsomal S-(+)-mephenytoin 4'-hydroxylation efficiently but was less efficient on R-(-)-mephenytoin 4'-hydroxylation in extensive metabolizers and on 4'-hydroxylation of mephenytoin enantiomers in poor metabolizers . The antibodies also inhibited R-(-)-hexobarbital 3' alpha-hydroxylation and S-(+)-hexobarbital 3' beta-hydroxylation but did not effectively inhibit the hydroxylation of the two other optical isomers of hexobarbital in extensive metabolizers and of four stereoisomers in poor metabolizers . These findings indicate the close relationship between polymorphic mephenytoin 4'-hydroxylation and two stereospecific hexobarbital hydroxylations, and they suggest that P-450 human-2 is a typical S-(+)-mephenytoin 4'-hydroxylase and a major hexobarbital 3'-hydroxylase in the livers of extensive metabolizers . The findings were further supported by the experiments that used P-450 human-2 complementary dexoyribonucleic acid-derived protein in yeast microsomes. Arch Biochem Biophys, 1990 Mar, 277(2), 368 - 73 Presence of the cytosolic factor stimulating the import of precursor of mitochondrial proteins in rabbit reticulocytes and rat liver cells; Ono H et al.; Previously we purified a cytosolic factor that stimulates the import of the extrapeptide (the synthetic peptide of the presequence of ornithine aminotransferase) into the mitochondrial matrix (Ono, H., and Tuboi, S., 1988, J . Biol . Chem . 263, 3188-3193) . In this work this cytosolic factor was shown also to stimulate the import of the precursors of ornithine aminotransferase, a large subunit of succinate dehydrogenase, and sulfite oxidase . The amounts of these precursors bound to the outer mitochondrial membrane were increased by this cytosolic factor, suggesting that the cytosolic factor participates in the recognition step in the import process of the precursor protein . When the cytosolic factor was applied to an ATP-agarose column, the import-stimulating activity was recovered entirely in the unadsorbed fraction . Immunochemical studies showed that in these conditions the 70-kDa heat shock-related protein (Hsp 70) was present exclusively in the fraction adsorbed to the ATP-agarose column . The cytosolic factor is thus different from the 70-kDa heat shock-related protein, which was identified as a factor required for the import of mitochondrial proteins in yeast . The cytosolic factor was also detected in the cytosol of rat liver cells, and a considerable amount of this factor was recovered from rat liver mitochondria by washing them with high salt buffer, suggesting that the cytosolic factor has affinity to the outer mitochondrial membrane and binds to its receptor on the membrane . From these results, we conclude that the cytosolic factor forms a complex with the precursor of mitochondrial protein and then this complex binds to the outer mitochondrial membrane, probably via the receptor of the cytosolic factor. Int J Dev Biol, 1990 Mar, 34(1), 93 - 109 Meiosis reinitiation as a model system for the study of cell division and cell differentiation; Guerrier P et al.; In this paper, we review our findings concerning the control of meiosis reinitiation in starfish oocytes and discuss recent advances that lead to characterization of the maturation promoting factor (MPF) responsible for G2-M transition . It is now agreed that appearance of this factor, which triggers nuclear envelope breakdown, chromosome condensation and metaphase spindle formation, corresponds to the activation of a M-phase specific H1-kinase . MPF has been shown to be constituted of equimolar amounts of a 34 kDa catalytic subunit protein homologous to the yeast cdc2/CDC28 gene product and a cyclin protein homologous to the yeast cdc13 gene product . "In vivo" and "in vitro" studies based on the use of inhibitors of protein synthesis, protein kinases, phosphoprotein phosphatases and proteases lead to a better understanding of the complex series of events which regulate activation and inactivation of MPF . In the unfertilized metaphase 2-arrested vertebrate oocyte, it has also been shown that stabilization of MPF depends on the kinase activity of the c-mos protooncogene . This review attempts to illustrate how the significant progress made in the understanding of the regulation of cell cycle transverse directly resulted from the convergence of observations in multidisciplinary studies in yeast genetics, development and oncogenesis . It also offers a model for considering the highly integrated events which, starting at the level of the plasma membrane, may eventually result in early cell differentiation. Mutagenesis, 1990 Mar, 5(2), 159 - 64 The use of historical data for identifying biologically unimportant but statistically significant results in genotoxicity assays; Mitchell ID et al.; The definition of a negative result is a problem in genetic toxicology . Here we suggest that a result may be considered biologically unimportant (negative) if it falls within the limits of variation usually found in the negative controls of the particular test . To determine 'usual' variation, we have set 95% confidence limits on three indices of variation, calculated from historical values for duplicate negative control data from several genotoxicity tests . These tests showed four characteristic types of response and the appropriate index of variability was determined for each . Where there was little test-to-test variation in true mean (micronucleus test and metaphase analysis), confidence limits set on the overall distribution of negative controls were the best index of variability . In other assays (Ames, yeast and mouse lymphoma), there was considerable test-to-test variation so that differences between, or ratios of, the members of control duplicates were the preferred measure of variability . This approach can define what is biologically unimportant in terms of the test system . However, no inference can be drawn as to potential importance . Thus the main use is the removal of the positive 'label' from statistically significant results which fall within the usual range of spontaneous variation for the assay under consideration. Rev Infect Dis, 1990 Mar-Apr, 12 Suppl 3, S334 - 7 Overview of studies of fluconazole in oropharyngeal candidiasis; Hay RJ; Studies with fluconazole in oropharyngeal candidiasis have focused primarily on three groups of infections: chronic atrophic candidiasis, oropharyngeal infections associated with either neutropenia or AIDS, and chronic mucocutaneous candidiasis . In two studies of chronic atrophic candidiasis associated with dentures, 82 patients received 7 or 14 days of therapy with fluconazole (50 mg daily) . Clinical and mycologic cure rates ranged from 69% to 100%, with the best results occurring with 14 days of therapy in combination with the cleansing of dentures . Thirteen patients with oroesophageal candidiasis associated with chronic mucocutaneous candidiasis were treated with 50 or 200 mg of fluconazole daily, and clinical and mycologic remissions were achieved in a mean period of 10 days . So far 95 patients have been treated with fluconazole for oropharyngeal candidiasis associated with malignancy, therapeutic immunosuppression, AIDS, or AIDS-related complex . Infection was cured by clinical criteria in 84% of those studied . While the majority of patients with clinical cure had significant reductions in the number of yeast colonies, only 48% had negative oral cultures at the end of therapy with courses of 50 mg of fluconazole daily for 5 days to 8 weeks. J Obstet Gynecol Neonatal Nurs, 1990 Mar-Apr, 19(2), 133 - 8 Symptoms of chronic vaginal infection and microscopic condyloma in women; Deitch KV et al.; The purpose of this study was to determine if women who self-reported symptoms of chronic vaginal infection might be at risk for microscopic condyloma and, therefore, in need of colposcopic examinations . The hypothesis tested was that women who report yeast-type symptom chronicity and who have yeast-negative laboratory studies will have a significant incidence of microscopic vulvar condyloma . The hypothesis was supported by the results . Findings show a need to broaden the use of the colposcope to detect condyloma in women with a chronicity of unexplained yeast-type symptoms. Infect Dis Clin North Am, 1990 Mar, 4(1), 29 - 46 Vaccines to prevent hepatitis B and hepatitis A virus infections; Hadler SC; Vaccines to prevent hepatitis B infection became available in 1982, and were recommended primarily for adults considered at high risk because of exposure in the workplace or lifestyles that lead to sexual or parenteral exposure to the virus . Plasma-derived and recombinant vaccines produced in yeast are highly immunogenic, safe, and effective for all age groups . Decreased response and protection by vaccine occur in older persons and in persons with immunosuppressive illnesses . The main issues concerning current vaccination programs include the possible use of lower doses to reduce the cost of the vaccine, the duration of protection and need for vaccine booster doses, and the need for postvaccination testing in hospitals . The failure of current vaccination programs to decrease disease incidence, a matter of great concern nationally, can be ascribed to failure to deliver vaccine to the groups at highest risk . More effective strategies for control, including the universal vaccination of infants or adolescents, must now be examined . Newer vaccines that incorporate other viral antigens and that may offer increased efficacy are being developed . Vaccines to prevent hepatitis A--both killed and live attenuated vaccines--are undergoing clinical trials and may become available in the next 5 years. Vaccine, 1990 Mar, 8 Suppl, S56 - 9; discussion S60-2 Comparison of a recombinant DNA hepatitis B vaccine alone or in combination with hepatitis B immune globulin for the prevention of perinatal acquisition of hepatitis B carriage; Poovorawan Y et al.; The protective efficacy of a recombinant DNA yeast-derived hepatitis B vaccine was assessed alone or in combination with hepatitis B immune globulin (HBIg) in neonates born to surface antigen (HBsAg)-positive and e antigen (HBeAg)-positive mothers . Neon