Mol Gen Genet, 1998 Jan, 257(2), 113 - 23 patufet, the gene encoding the Drosophila melanogaster homologue of selenophosphate synthetase, is involved in imaginal disc morphogenesis; Alsina B et al.; Proliferation in imaginal discs requires cell growth and is linked to patterning processes controlled by secreted cell-signalling molecules . To identify new genes involved in the control of cell proliferation we have screened a collection of P-lacW insertion mutants that result in lethality in the larval/pupal stages, and characterized a novel gene, patufet (ptuf) . Inactivation of ptuf by a P element insertion in the 5' untranslated region leads to aberrant imaginal disc morphology characterized by a reduction in mass of discs and disorganization of disc cells where no folding or patterning can be detected . Moreover, apoptotic cells can be observed in these small and abnormal mutant discs . To examine the role of ptuf we have studied its clonal behaviour in genetic mosaics generated by mitotic recombination . The mutation causes reduced cell viability, smaller cell size and stops vein differentiation . Non-autonomous effects, such as abnormal differentiation of wild-type cells surrounding the clones, are also observed . We have cloned the ptuf gene of Drosophila melanogaster and found that it encodes a selenophosphate synthetase, which is the first identified in insects . Mutant flies transformed with the full-length cDNA show complete reversion of lethality and disc phenotype . Northern blot analysis and in situ hybridization indicate that the ptuf gene is expressed in imaginal discs as well as at different stages of development . The synthesis of selenoproteins by the selenophosphate synthetase, the role of selenoproteins in the maintenance of the oxidant/antioxidant balance of the cell and its possible implications in imaginal disc morphogenesis are discussed. FEBS Lett, 1998 Jan 30, 422(2), 231 - 4 The cytochrome subunit structure in the photosynthetic reaction center of Chromatium minutissimum; Chamorovsky SK et al.; Gel-electrophoretic assay revealed that the photosynthetic reaction center (RC) of Chromatium minutissimum, in contrast to the well-known RC Rhodopseudomonas viridis, consists of five rather than four subunits with molecular masses of 37, 34, 25, 19, and 17 kDa . The 37- and 19-kDa subunits are stained with tetramethylbenzidine for the cytochrome c hemes . Absorption spectra show that the concentration of reduced cytochromes in the C . minutissimum RC poised at redox potential of -150 mV (fully reduced pool of hemes) is about three times more than in the C . minutissimum RC poised at redox potential of +260 mV (only high-potential hemes are reduced) . The results of redox titration of absorption changes at the cytochrome c alpha-band are most appropriately approximated by a six-component theoretical curve with the midpoint potentials of Em1 = 390 mV, Em2 = 320 mV, Em3 = 210 mV, Em4 = 100 mV, Em5 = 20 mV, and Em6 = -50 mV . Possible functions of the cytochromes with the midpoint potentials 210 and 100 mV, which have not been found in purple bacteria before, are discussed. Mol Microbiol, 1998 Feb, 27(3), 661 - 7 The requirement for DsbA in pullulanase secretion is independent of disulphide bond formation in the enzyme; Sauvonnet N et al.; Results from previous studies have suggested that an intramolecular disulphide bond in the exoprotein pullulanase is needed for its recognition and transport across the outer membrane . This interpretation of the data is shown here to be incorrect: pullulanase devoid of all potential disulphide bonds is secreted with apparently the same efficiency as the wild-type protein . Furthermore, the periplasmic disulphide bond, oxidoreductase DsbA, previously shown to catalyse the formation of a disulphide bond in pullulanase and to decrease its transit time in the periplasm, is shown here to be required for the rapid secretion of pullulanase devoid of disulphide bonds . Several possible explanations for the role of DsbA in pullulanase secretion are discussed. Dev Biol, 1998 Jan 15, 193(2), 115 - 26 A molecular analysis of hyalin--a substrate for cell adhesion in the hyaline layer of the sea urchin embryo; Wessel GM et al.; The hyaline layer of echinoderm embryos is an extraembryonic matrix that functions as a substrate for cell adhesion through early development . The major constituent of the hyaline layer is the protein hyalin, a fibrillar glycoprotein of approximately 330 kDa that multimerizes in the presence of calcium . Here we provide a molecular characterization of hyalin and identify a region of the protein that is important for its function in cell adhesion . Partial hyalin cDNAs were identified from two sea urchin species, Strongylocentrotus purpuratus and Lytechinus variegatus, by screening expression libraries with monoclonal antibodies to hyalin . The cDNAs each encode a tandemly arranged series of conserved repeats averaging 84 amino acids . These hyalin repeats are as similar between the two species as they are to repeats within each species, suggesting a strong functional conservation . Analysis of this repeat shows that it is a unique sequence within the GenBank database with only weak similarity to mucoid protein sequences . The hyalin mRNA is approximately 12 kb in length and is present in developing oocytes coincident with the appearance of cortical granules, the vesicle in which the hyalin protein is specifically packaged . The mRNA is present throughout oogenesis but is rapidly lost at oocyte maturation so that eggs and early embryos have no detectable hyalin mRNA . The hyalin protein, however, remains at relatively constant levels throughout development . Thus, all the hyalin protein present during early development, when no RNA is detectable, is maternally derived and exocytosed from cortical granules at fertilization . Hyalin mRNA reaccumulates in embryos beginning at the mesenchyme blastula stage; a RNA gel blot and in situ hybridization analysis of gastrulae and larvae shows a progressive confinement of hyalin mRNA to the aboral ectoderm . Recombinant hyalin containing the tandem repeat region of the protein was expressed in bacteria and is shown to serve as an adhesive substrate, almost equal to that of native hyalin, in cell adhesion assays . This adhesive activity is partially blocked by dilute hyalin monoclonal antibody Tg-HYL to the same extent as that for native hyalin . Thus, this hyalin repeat region appears to contain the ligand for the hyalin cell surface receptor . These data help explain some of the classic functions ascribed to the hyalin protein in early development and now enable investigators to focus on the mechanisms of cell interactions with the hyaline layer . Blood, 1998 Feb 15, 91(4), 1438 - 45 Effects of increased anionic charge in the beta-globin chain on assembly of hemoglobin in vitro; Adachi K et al.; Studies on assembly in vitro of alpha-globin chains with recombinant beta16 Gly-->Asp, beta95 Lys-->Glu, beta120 Lys-->Glu and beta16 Gly-->Asp, 120 Lys-->Glu human beta-globin chain variants in addition to human betaA- and betaS-globin chains were performed to evaluate effects of increased anionic charge in the beta chain on hemoglobin assembly using soluble recombinant beta-globin chains expressed in bacteria . A beta112 Cys-->Asp change was also engineered to monitor effects on assembly of increased negative charge at alpha1beta1 interaction sites . Order of tetramer formation in vitro under limiting alpha-globin chain conditions showed Hb betaG16D, K120E = Hb betaK120E = Hb betaK95E > Hb betaG16D > Hb A > Hb S >>> Hb betaC112D . In addition, beta112 Cys-->Asp chains exist as monomers rather than beta4 tetramers in the absence of alpha chains, and the beta chain in Hb betaC112D tetramers was readily exchanged by addition of betas . These results suggest that affinity between alpha and beta chains is promoted by negatively-charged beta chains up to a maximum of two additional net negative charges and is independent of location on the surface except at the alpha1beta1 interaction site . In addition, our findings show that beta112 Cys on the G helix is critical for facilitating formation of stable alphabeta dimers, which then form functional hemoglobin tetramers, and that beta112 Cys-->Asp inhibits formation of stable alpha1beta1 and beta1beta2 interactions in alpha2beta2 and beta4 tetramers, respectively. J Mol Evol, 1998 Jan, 46(1), 84 - 101 Evolution of substrate specificities in the P-type ATPase superfamily; Axelsen KB et al.; P-type ATPases make up a large superfamily of ATP-driven pumps involved in the transmembrane transport of charged substrates . We have performed an analysis of conserved core sequences in 159 P-type ATPases . The various ATPases group together in five major branches according to substrate specificity, and not according to the evolutionary relationship of the parental species, indicating that invention of new substrate specificities is accompanied by abrupt changes in the rate of sequence evolution . A hitherto-unrecognized family of P-type ATPases has been identified that is expected to be represented in all the major phyla of eukarya. Mol Cell Biol, 1998 Mar, 18(3), 1163 - 71 Functional relationships of Srb10-Srb11 kinase, carboxy-terminal domain kinase CTDK-I, and transcriptional corepressor Ssn6-Tup1; Kuchin S et al.; The Srb10-Srb11 protein kinase of Saccharomyces cerevisiae is a cyclin-dependent kinase (cdk)-cyclin pair which has been found associated with the carboxy-terminal domain (CTD) of RNA polymerase II holoenzyme forms . Previous genetic findings implicated the Srb10-Srb11 kinase in transcriptional repression . Here we use synthetic promoters and LexA fusion proteins to test the requirement for Srb10-Srb11 in repression by Ssn6-Tup1, a global corepressor . We show that srb10delta and srb11delta mutations reduce repression by DNA-bound LexA-Ssn6 and LexA-Tup1 . A point mutation in a conserved subdomain of the kinase similarly reduced repression, indicating that the catalytic activity is required . These findings establish a functional link between Ssn6-Tup1 and the Srb10-Srb11 kinase in vivo . We also explored the relationship between Srb10-Srb11 and CTD kinase I (CTDK-I), another member of the cdk-cyclin family that has been implicated in CTD phosphorylation . We show that mutation of CTK1, encoding the cdk subunit, causes defects in transcriptional repression by LexA-Tup1 and in transcriptional activation . Analysis of the mutant phenotypes and the genetic interactions of srb10delta and ctk1A suggests that the two kinases have related but distinct roles in transcriptional control . These genetic findings, together with previous biochemical evidence, suggest that one mechanism of repression by Ssn6-Tup1 involves functional interaction with RNA polymerase II holoenzyme. Infect Immun, 1998 Mar, 66(3), 1261 - 4 Identification of epitopes of fibronectin attachment protein (FAP-A) of Mycobacterium avium which stimulate strong T-cell responses in mice; Holsti MA et al.; The T-cell response to fibronectin attachment protein (FAP-A) in BALB/c and B10.BR mice was examined . Both strains developed strong T-cell responses to FAP-A, directed to single, unique epitopes . T cells from mice infected with Mycobacterium avium responded to FAP-A, suggesting a possible role in a protective immune response. Infect Immun, 1998 Mar, 66(3), 1023 - 7 Evidence for specific secretion rather than autolysis in the release of some Helicobacter pylori proteins; Vanet A et al.; We investigated whether Helicobacter pylori cells actively secrete proteins such as the urease subunits UreA and UreB and the GroES and GroEL homologs HspA and HspB or whether these proteins were present in the extracellular compartment as a consequence of autolysis . Using a subcellular fractionation approach associated with quantitative Western blot analyses, we showed that the supernatant protein profiles were very different from those of the cell pellets, even for bacteria harvested in the late growth phase; this suggests that the release process is selective . A typical cytoplasmic protein, a beta-galactosidase homolog, was found exclusively associated with the pellet of whole-cell extracts, and no traces were found in the supernatant . In contrast, UreA, UreB, HspA, and HspB were mostly found in the pellet but significant amounts were also present in the supernatant . HspA and UreB were released into the supernatant at the same rate throughout the growth phase (3%), whereas large portions of HspB and UreA were released during the stationary phase (over 30 and 20%, respectively) rather than during the early growth phase (20% and 6, respectively) . The profiles of protein obtained after water extraction of the bacteria with those of the proteins naturally released within the liquid culture supernatants demonstrated that water extraction led to the release of a large amount of protein due to artifactual lysis . Our data support the conclusion that a specific and selective mechanism(s) is involved in the secretion of some H . pylori antigens . A programmed autolysis process does not seem to make a major contribution. J Endod, 1998 Jan, 24(1), 11 - 4 Recontamination of coronally unsealed root canals medicated with camphorated paramonochlorophenol or calcium hydroxide pastes after saliva challenge; Siqueira Junior JF et al.; This in vitro study evaluated the ability of some medications to prevent recontamination of coronally unsealed root canals by bacteria from saliva . The medications tested were camphorated paramonochlorophenol (CPMC) applied in cotton pellets in the pulp chamber; calcium hydroxide/saline solution paste filling the root canal; and calcium hydroxide/CPMC/glycerin paste also filling the root canal . Medicated canals were exposed to saliva, and the number of days required for total recontamination to occur was recorded . Canals medicated with CPMC in cotton pellets were thoroughly recontaminated within an average of 6.9 days . Canals filled with calcium hydroxide/saline solution and calcium hydroxide/CPMC/glycerin showed entire recontamination within an average of 14.7 and 16.5 days, respectively . Calcium hydroxide pastes were significantly more effective than CPMC (p < 0.05). East Afr Med J, 1997 Aug, 74(8), 519 - 22 Socio-biological status of Nigerian males with primary and secondary infertility; Alemnji GA et al.; Husbands in 100 consecutive couples complaining of lack of pregnancy after one year of normal intercourse were engaged in this study . Information from a structured questionnaire administered to these 100 men showed that 46% had primary infertility (had never impregnated any woman) and 54% secondary infertility (had in the past impregnated at least one woman irrespective of the outcome of the pregnancy) . The mean ages (years) and standard error of mean for the primary and secondary infertile groups were 33.46 +/- 1.45 and 39.28 +/- 1.41 respectively . The difference was statistically significant (p < 0.05) . Semen culture for growth of bacteria was positive in 59.3% of subjects with secondary infertility as opposed to 40.7% for primary infertility . The difference was, again, statistically significant (p < 0.05) . These findings indicate that a higher proportion of husbands in infertile couples in a group of this environment had secondary infertility, were older and were more likely to harbour infections in their semen than those with primary infertility . Hence there should be a greater awareness of the significant involvement of bacterial infection of the genital tract of infertile Nigerian subjects than and before this factor should be taken into account in the prevention and treatment strategies for infertility in this and presumably other tropical countries. FEMS Microbiol Lett, 1998 Feb 1, 159(1), 137 - 44 Macrorestriction analysis of Desulfurella acetivorans and Desulfurella multipotens; Pradella S et al.; The genomes of the phylogenetically and physiologically unique bacteria Desulfurella acetivorans DSM 5264T and D . multipotens DSM 8415T were characterized and compared by pulsed field gel electrophoresis (PFGE) . Macrorestriction patterns made of large PFGE separated DNA fragments were generated by digesting the genomic DNAs of both strains with the rare cutting restriction endonucleases ApaI, AscI, EagI, RsrII, SacII, SalI as well as with the intron encoded endonuclease I-CeuI . The sum of calculated fragment sizes from digests of the first six enzymes yielded estimates for the chromosome sizes of D . acetivorans with a mean of 1939.0 +/- 26.0 kb and for D . multipotens with a mean of 1864.0 +/- 23.0 kb . Within the patterns obtained from EagI and RsrII cleavages the apparent differences could be attributed to DNA insertion or deletion and to point mutation . The single, circular chromosomes of the two strains contain two copies of 23S rRNA genes each . Different extrachromosomal elements were detected in both strains. Mol Microbiol, 1998 Jan, 27(2), 323 - 36 Evidence for pore-forming ability by Legionella pneumophila; Kirby JE et al.; Legionella pneumophila is the cause of Legionnaires' pneumonia . After Internalization by macrophages, it bypasses the normal endocytic pathway and occupies a replicative phagosome bound by endoplasmic reticulum . Here, we show that lysis of macrophages and red blood cells by L . pneumophila was dependent on dotA and other loci known to be required for proper targeting of the phagosome and replication within the host cell . Cytotoxicity occurred rapidly during a high-multiplicity infection, required close association of the bacteria with the eukaryotic cell and was a form of necrotic cell death accompanied by osmotic lysis . The differential cytoprotective ability of high-molecular-weight polyethylene glycols suggested that osmotic lysis resulted from insertion of a pore less than 3 nm in diameter into the plasma membrane . Results concerning the uptake of membrane-impermeant fluorescent compounds of various sizes are consistent with the osmoprotection analysis . Therefore, kinetic and genetic evidence suggested that the apparent ability of L . pneumophila to insert a pore into eukaryotic membranes on initial contact may play a role in altering endocytic trafficking events within the host cell and in the establishment of a replicative vacuole. Plant Mol Biol, 1998 Feb, 36(3), 427 - 37 cDNA cloning, substrate specificity and expression study of tobacco caffeoyl-CoA 3-O-methyltransferase, a lignin biosynthetic enzyme; Martz F et al.; Four caffeoyl-CoA 3-O-methyltransferase (CCoAOMT) cDNA clones were isolated from RNA extracted from TMV-infected tobacco leaves using an heterologous DNA probe . The cDNAs were 84-93% identical in their nucleotide sequences, indicating that they are the products of four closely related genes . A comparison of the CCoAOMT cDNAs with database sequences and Southern blot analysis indicated that they are encoded by a new CCoAOMT family of tobacco . Overall expression of this gene family in tobacco tissues was investigated by RNA blot analysis . The expression of each individual gene was studied by RT-PCR coupled with RFLP analysis of PCR products, taking advantage of the presence of specific restriction sites in each cloned cDNA . Two members of the CCoAOMT gene family appeared to be constitutively expressed in various plant organs and tissues whereas the two others were preferentially expressed in flower organs, after tobacco mosaic virus (TMV) infection or elicitor treatment of leaves . The CCoAOMT enzymatic protein expressed in bacteria was purified and shown to be specific for the caffeoyl-CoA and 5-hydroxyferuloyl-CoA esters and to have no activity against free caffeic acid and 5-hydroxyferulic acid . The pattern of CCoAOMT transcript accumulation during development of tobacco stem was found closely related to that of COMT I genes which have been shown to be specifically involved in lignin biosynthesis . Moreover, the inhibition of COMT I gene expression in transgenic tobacco was also shown to decrease CCoAOMT gene expression, particularly in the most lignified tissues . Thus, the expression pattern and the substrate specificity of tobacco CCoAOMT sustain a preferential role in lignin biosynthesis. Plant Mol Biol, 1998 Feb, 36(3), 353 - 63 Characterization and transcriptional regulation of the Synechocystis PCC 6803 petH gene, encoding ferredoxin-NADP+ oxidoreductase: involvement of a novel type of divergent operator; van Thor JJ et al.; The petH gene, encoding ferredoxin-NADP+ oxidoreductase (FNR), has been characterised in the unicellular cyanobacterium Synechocystis PCC 6803 . Its product, FNR, was heterologously produced and functionally characterized . The start-site of the monocystronic petH transcript was mapped 523 bp upstream of the predicted PetH initiation codon, resulting in an unusually large 5'-untranslated region . The 5' end of the petH transcript is situated within the open reading frame of phosphoribulokinase (encoded by prk), which is transcribed in opposite orientation with respect to petH . The transcription start site of the prk transcript was mapped 219 bp upstream of the initiation codon, resulting in a 223 bp antisense region between both transcripts . Under many conditions the expression of both genes (i.e . petH and prk) is co-regulated symmetrically at the transcriptional level, as was concluded from both northern hybridization experiments and from primer extension analyses; it became uncoupled, however, when specifically petH expression was stimulated, independent of prk expression, by stressing the Synechocystis cells with high salt concentrations . A model for a new type of bidirectional operator, regulating the expression of petH and prk, is proposed. Oper Dent, 1997 Jul-Aug, 22(4), 149 - 58 Biocompatability of compomer restorative systems on nonexposed dental pulps of primate teeth; Tarim B et al.; This study evaluated the histologic response of total-etched and nonetched compomer restored cavity preparations . One hundred fifteen class 5 cavity preparations were placed in the teeth of four healthy adult monkeys at 7, 27, and 90 days . A 37% H3PO4 was applied for 10 seconds and rinsed in total-etched preparations . No statistical differences were seen in inflammatory reactions among total-etched or nonetched compomers at 7, 27, and 90 days . There were no statistical differences in inflammatory cell responses among all compomer systems in regard to time intervals . Pulpal responses of compomers were greater than IRM at each time period . Pulp responses were associated with stained bacteria in 32 of 89 compomer teeth . No necrotic pulps were seen in any teeth . Statistical data show a positive correlation (P < 0.05) between bacterial presence and pulpal inflammation . IRM pulps showed no inflammation or bacterial staining . Compomers are biologically compatible with pulp tissues when bacteria are excluded. J Clin Laser Med Surg, 1996 Feb, 14(1), 7 - 11 Long-term clinical evaluation of endodontically treated teeth by Nd:YAG lasers; Gutknecht N et al.; It was possible for the patients to avoid surgical intervention in a number of complicated periapical endodontic situations by means of Nd:YAG laser-assisted sterilization . A WSR has only very good primary results and the long-term successes are very limited . Once a lesions has healed in the manner explained in this study, in other words, with regeneration of the periapical anatomy, there is a very good long-term prognosis . Laser technology is an instrument whose overall effects represent a decisive improvement in the efficiency of conservative endodontic treatment in fields that were previously outside our sphere of influence. Vojnosanit Pregl, 1997 Nov-Dec, 54(6), 541 - 8 {Physical therapy in the rehabilitation of patients with aerobic infections of the extremities}; Durovic A et al.; OBJECTIVE: To evaluate how infection of extremity after war wound influenced the possibilities and immediate effects of a physical therapy . METHODS: The retrospective clinical investigation comparing two groups: group A (n = 86) with infection, group B (n = 87) without infection . Main indicators for possibilities of the physical therapy were the numbers and types of physical procedures used . For the estimation of immediate effects of physical therapy the muscle power and the range of motion were used . RESULTS: The number of daily physical procedures in the group with infection, compared to the group without infection, was significantly lesser ((A: 2.87 +/- 1.73; B: 4.02 +/- 1.73; p < 0.001) . The patients with infection were significantly less frequently submitted to thermotherapy, hydrotherapy, interferent current and electrostimulation . Patients with infection, compared to patients without infection, had significantly poorer improvement of amplitude of analyzed movements at the end of treatment (A: 6.66 +/- 7.28 degrees; B: 16.66 +/- 14.79 degrees; p < 0.001) . CONCLUSION: The infection of the extremities limited the possibilities and reduced the immediate effects of physical therapy. Nephrol Dial Transplant, 1998 Jan, 13(1), 130 - 3 Successful use of single-lumen, urokinase immobilized femoral catheters as a temporary access for haemodialysis; Takeda K et al.; METHODS: Placement of a femoral vein catheter as a temporary vascular access for haemodialysis was conducted and the indications, catheter patency rate, and incidence of catheter-related infections were examined . An urokinase immobilized femoral vein catheter (UIFC) is a soft polyurethane single-lumen catheter 2.7 mm in diameter and 22 cm in length which needs no heparin infusion (Japan Shawood Co., Ltd., Tokyo; Unitica Co., Ltd., Hyogo, Japan) . A soft silicon rubber was attached to the tip of the catheter in order to avoid excessive bleeding during insertion . Aseptic adhesive wound dressing was employed at the exit-site which was cleansed with popidone-iodine and renewed at each dialysis session . RESULTS: Eighty-one UIFCs were used for haemodialysis in 64 patients (acute renal failure: 11; vascular access trouble: 53; initiation of chronic dialysis: 17) . The average age of the patients was 58 +/- 13 years, ranging from 26 to 80 years . The mean duration of catheter indwelling was 22.4 +/- 13.1 days . An adequate blood flow of 180-200 ml/min was obtained through UIFC and returned to another peripheral vein punctured at each dialysis session . Unexplained fever occurred in four cases while the UIFC was in place (4.9%) but culture of either blood or the catheter tip was negative for bacteria . The catheter was removed immediately and fever subsided in all cases . The overall catheter survival rate was 84% at 34 days calculated using the Kaplan-Meier method . Catheter insertion was easy to perform and no serious complications such as pulmonary embolism or septicaemia occurred . CONCLUSION: Our modified type of UIFC is very useful as a temporary access for haemodialysis with a very low incidence of catheter-related infections and no need for heparinization . Excellent catheter patency was maintained with the plug system and careful dressing techniques without unnecessary bleeding during catheter care. Nutr Rev, 1998 Jan, 56(1 Pt 1), 1 - 8 Human adult-onset lactase decline: an update; Lee MF et al.; Human adult-onset lactase decline is a biologic feature characteristic of the maturing intestine in the majority of the world's population . The digestion and absorption of lactose, the major carbohydrate in milk and also the main substrate for lactase, is often variable, a consequence of lactase levels, gastric emptying rate, and colonic salvage . Although commercially available "lactase" products alleviate symptoms in many lactose-intolerant people, a greater understanding of this variability in lactose tolerance could lead to interventions that reduce the rate of gastric emptying and/or increase the proliferation of lactose-metabolizing bacteria in the colon, leading to more efficient lactose utilization . Adult-onset lactase decline appears to be a risk factor for developing osteoporosis, owing to avoidance of dairy products or interference of undigested lactose with calcium absorption . Elderly with both adult-onset lactase decline and atrophic gastritis or those undergoing anti-ulcer treatment may have an increased risk of low calcium absorption owing to the lack of gastric acid that facilitates calcium uptake . Thus, lactose-intolerant elders should monitor their calcium nutrition status carefully. Przegl Lek, 1997, 54(7-8), 561 - 4 {Some aspects of ozone therapy}; Antoszewski Z et al.; Authors present ozone-biochemistry, describe ozone production and application method of oxygen-ozone mixture . Authors describe application pf ozone therapy to treatment of virus or bacteria diseases and others as well as contraindications to the ozone therapy, side effects . Authors evaluate also effects of ozone therapy. Mikrobiol Z, 1997 Sep-Oct, 59(5), 13 - 21 {The chemical composition and biological activity of the glyco polymers in Ralstonia solanacearum (Yabuchi et al., 1995)}; Varbanets LD et al.; Lipopolysaccharide and extracellular glyco polymers were isolated from cells of phytopathogenic bacteria Ralstonia solanacearum . It was established that the O-specific polysaccharides are characterized by a linear structure, composed of tetrasaccharide repeating units containing three L-rhamnose residues and one of N-acetylglucosamine residue . Core oligosaccharide along with typical monosaccharides such as rhamnose, glucose, heptose, KDO and glucosamine included fucose, galactose and arabinose . Serological activity of lipopolysaccharide is due to both O-specific polysaccharide and lipid A component . Extracellular glyco polymers contained both neutral (GP 1) and charged (GP 2 and GP 3) components . Rhamnose and glucose (GP 1, GP 2) or galactose (GP 3) were predominant monosaccharides, GP 2 displayed a high phytotoxic action in respect to tomato plants. Br Dent J, 1998 Jan 10, 184(1), 33 - 8 Is there a link between periodontal disease and coronary heart disease? Seymour RA, Steele JG. OBJECTIVE: To provide a critical review of the studies completed to date that have investigated a link between coronary heart disease and dental health . DESIGN: Retrospective analysis . SETTING: Mainly hospital-based patients or subjects involved in longitudinal health care studies . MAIN OUTCOME MEASURES: The incidence of coronary heart disease and its relationship to dental health and other recognised risk factors . RESULTS: Evidence suggests that dental health, in particular periodontal disease, may be a significant risk factor for coronary heart disease and further coronary events . Possible biological mechanisms that link the two diseases are appraised . CONCLUSIONS: There does appear to be increasing evidence that a relationship exists between dental health and coronary heart disease, especially in males aged 40-50 years . The presence of a hyperinflammatory monocyte phenotype may provide a common biological mechanism that links the two diseases. Br Dent J, 1998 Jan 10, 184(1), 12 - 6 Advances in periodontal diagnosis . 1 . Traditional clinical methods of diagnosis; Eley BM et al.; This series will consider the limitations of traditional periodontal diagnostic techniques and the recent advances which have sought to overcome them . It will cover improvements in probing and radiographic techniques and also discusses the attempts to find bacterial or tissue-derived markers which will diagnose or predict periodontal disease activity . The first paper describes the traditional methods of periodontal diagnosis. J Clin Gastroenterol, 1997, 25 Suppl 1, S149 - 63 Helicobacter pylori factors involved in the development of gastroduodenal mucosal damage and ulceration; Figura N; Many putative virulence determinants of Helicobacter pylori are believed to trigger and worsen the gastroduodenal mucosa damage observed in infected patients . H . pylori urease reacts with the gastric urea and generates ammonia; ammonia combines with water and yields ammonium hydroxide, which is cytotoxic . Ammonia may also inhibit cell proliferation and cause indirect mucosal injury by stimulating neutrophils . Phospholipases may damage the gastric mucosa by degrading phospholipids and generating precursors of ulcerogenic components . Other enzymes, such as protease, neuraminidase, fucosidase, and alcohol dehydrogenase, can contribute to damage of the gastric epithelium by destroying the integrity of mucus or by inducing lipid peroxidation . Infection by vacuolating cytotoxic (VacA+) H . pylori strains is considered to constitute increased risk for development of peptic ulcer and gastric cancer . Exploration of the vacA gene structure has shown the existence of strongly toxigenic strains, and has confirmed at the molecular level the increased ulcerogenic potential of VacA+ H . pylori strains . A pathogenicity island called cag has been recently described in Type 1 H . pylori strains (VacA+/CagA+).cag contains the cagA gene (whose expression is associated with toxigenicity) and many genes, some of which are highly homologous to virulence genes of other virulent bacteria, that account for the enhanced pathogenic potential of CagA+ organisms. Immunol Res, 1998, 17(1-2), 269 - 78 Immunobiology of interleukin-12; Trinchieri G; Interleukin-12 (IL12) is a heterodimeric cytokine which is produced by phagocytic cells and antigen-presenting cells within a few hours of infection, particularly in the case of bacteria and intracellular parasites, and acts as a proinflammatory cytokine, activating natural killer (NK) cells, and, through its ability to induce interferon-gamma(IFN gamma) production, enhancing the phagocytic and bacteriocidal activity of phagocytic cells and their ability to release proinflammatory cytokines, including IL12 itself . Furthermore, IL12 produced during the early phases of infection and inflammation, sets the stage for the ensuing antigen-specific immune response, favoring differentiation and function of T helper type 1 (Th1) T cells while inhibiting the differentiation of Th2 T cells . Thus, IL12, in addition to being a potent proinflammatory cytokine, is a key immunoregulator molecule in Th1 responses. J Biol Chem, 1998 Jan 30, 273(5), 2874 - 84 Efficient generation of major histocompatibility complex class I-peptide complexes using synthetic peptide libraries; Stevens J et al.; The use of synthetic random peptide libraries is a powerful technology for the study of many aspects of antigen presentation and peptide selection by major histocompatibility complex (MHC) molecules . Here we have used them in conjunction with a recombinant system to determine the peptide binding motifs of three classical class I MHC molecules of the laboratory rat: RT1-Aa, RT1-Au, and RT1-A1c . Described is a method for producing large amounts of soluble class I heavy and light chains in bacteria . Refolding RT1-Aa heavy chain (HC) with rat beta2-microglobulin (beta2m) in the presence of a specific peptide and the subsequent purification of the complex yielded conformationally correct material . This was assessed by gel chromatography, SDS-polyacrylamide gel electrophoresis, isoelectric focussing gel electrophoresis, enzyme-linked immunosorbent assay, and fluorescence-activated cell sorter analysis employing a previously unreported method utilizing a His-Tag affinity silica . By refolding RT1-Aa HC and rat beta2m around a random nonapeptide library and subjecting the resulting complex to acid elution of the bound peptides and pool sequencing, the peptide binding motif for this MHC class I molecule was determined . Results corresponded well with those previously determined from naturally bound peptides and in addition gave a clear and unambiguous signal for the C-terminal anchor residue . This method was then applied to determine the previously undescribed binding motifs for RT1-Au and RT1-A1c . For both molecules, the whole motif was confirmed from naturally bound peptides . We propose this method as an alternative way to obtain the whole class I MHC peptide motif, particularly when a specific antibody is unavailable and/or natural expression of the class I molecule of interest is low. EMBO J, 1998 Jan 2, 17(1), 170 - 80 AGO1 defines a novel locus of Arabidopsis controlling leaf development; Bohmert K et al.; An allelic series of the novel argonaute mutant (ago1-1 to ago1-6) of the herbaceous plant Arabidopsis thaliana has been isolated . The ago1 mutation pleotropically affects general plant architecture . The apical shoot meristem generates rosette leaves and a single stem, but axillary meristems rarely develop . Rosette leaves lack a leaf blade but still show adaxial/abaxial differentiation . Instead of cauline leaves, filamentous structures without adaxial/abaxial differentiation develop along the stem and an abnormal inflorescence bearing infertile flowers with filamentous organs is produced . Two independent T-DNA insertions into the AGO1 locus led to the isolation of two corresponding genomic sequences as well as a complete cDNA . The AGO1 locus was mapped close to the marker mi291a on chromosome 1 . Antisense expression of the cDNA resulted in a partial mutant phenotype . Sense expression caused some transgenic lines to develop goblet-like leaves and petals . The cDNA encodes a putative 115 kDa protein with sequence similarity to translation products of a novel gene family present in nematodes as well as humans . No specific function has been assigned to these genes . Similar proteins are not encoded by the genomes of yeast or bacteria, suggesting that AGO1 belongs to a novel class of genes with a function specific to multicellular organisms. Curr Microbiol, 1998 Jan, 36(1), 9 - 12 Possible involvement of cysteine and histidine residues in the (NH4+ + Na+)-activated ATPase of an anaerobic alkaliphile, Amphibacillus xylanus; Okano Y et al.; Effect of various inhibitors on the (NH4+ + Na+)-activated ATPase of an anaerobic alkaliphile, Ep01(a strain of Amphibacillus xylanus), was examined . Among the chemicals tested, the enzyme was drastically inactivated by p-chloromercuribenzoic acid and diethyl pyrocarbonate . The ATPase activity of the enzyme, which was inactivated by p-chloromercuribenzoic acid and diethyl pyrocarbonate, was remarkably restored by beta-mercaptoethanol and hydroxylamine, respectively, suggesting the involvement of cysteine and histidine residues in the enzyme activity . Analysis of the inhibition kinetics by diethyl pyrocarbonate indicated that modification of a single histidine residue per ATPase molecule was sufficient to inactivate the enzyme. J Cell Sci, 1998 Jan, 111 ( Pt 1), 141 - 8 Degradation of phagosomal components in late endocytic organelles; Tjelle TE et al.; Phagosomes are formed when phagocytic cells ingest particles such as bacteria, viruses or synthetic beads of different kinds . The environment within the phagosome gradually changes to generate degradative conditions . These changes require multiple interactions between the maturing phagosomes and the endocytic and the biosynthetic pathway . The phagosomes probably communicate with endocytic organelles by a transient fusion event, often referred to as the 'kiss-and-run' hypothesis . We have studied the role of endocytic organelles in the phagocytic pathway of J774 cells, a mouse macrophage cell line . We have used magnetic Dynabeads coated with 125ITC-IgG and 125ITC-OVA as phagocytic probes and were able to isolate the phagosomal fraction by means of a magnet . To separate lysosomes from other organelles in the endocytic pathway we allowed the cells to endocytose a pulse of colloidal gold particles complexed with ovalbumin . By combining this density shift technique with subcellular fractionation of a postnuclear supernatant in Percoll gradients we could isolate three endocytic fractions corresponding to early endosomes (the light Percoll fraction), late endosomes (the dense Percoll fraction) and lysosomes (the gold fraction) . We observed that the proteins linked to the ingested beads are initially cleaved in the phagosomes . This cleavage is inhibited by leupeptin, a thiol-protease inhibitor, and requires an acidic environment . However, efficient communication between the phagosomes and the endocytic pathway leads to the transfer of dissociated phagocytosed peptides of different sizes to late endosomes and lysosomes for further processing . Consequently, the late endosomes and the lysosomes may be involved in the degradation of phagocytosed compounds. Arch Histol Cytol, 1997 Dec, 60(5), 493 - 502 Demonstration and organization of duct-associated lymphoid tissue (DALT) of the main excretory duct in the monkey parotid gland; Matsuda M et al.; Duct-associated lymphoid tissue (DALT) of the main excretory duct in the monkey parotid gland was first demonstrated by light microscopy and by transmission and scanning electron microscopy . The DALT included a follicular area, a parafollicular area and a specialized overlying epithelium with distinct fine-structural elements . There was usually a solitary lymphoid follicle located in the subepithelial area near the orifice of the parotid duct . The lymphoid follicles typically had a distinct germinal center . Numerous immune cells often infiltrated into the epithelium overlying the lymphoid follicle . The superficial epithelial cells of the DALT were larger and flatter than the ordinary duct epithelial cells, and had short irregular microvilli on their luminal surface . They were also in close contact with immune cells such as dendritic cells and lymphocytes . Goblet cells were rare in this area . In addition, bacteria, seen at the duct orifice, were sometimes taken up by the flattened epithelial cells near the orifice . Latex microspheres administrated as particulate antigens at the duct orifice were selectively taken up by the flattened epithelial cells and also by the intraepithelial dendritic cells of the DALT . These morphological findings suggest that the epithelial cells of the DALT in parotid glands take up antigens from the duct lumen and transport them to adjacent immune cells, and that the DALT in parotid glands may serve as one of the inductive sites in the common mucosal immune system. Immunopharmacology, 1997 Dec, 38(1-2), 207 - 13 Complement factor I deficiency in a family with recurrent infections; Leitao MF et al.; Factor I deficiency causes a permanent, uncontrolled activation of the alternative pathway resulting in an increased turnover of C3 and consumption of factor B, factor H and properdin . Factor I deficiency is clinically associated with recurrent bacterial infections already in early infancy, mainly affecting the upper and lower respiratory tract, or presenting as meningitis or septicemia . We here report on a Brazilian family (n = 9) with known consanguinity, where in 3/7 children, suffering from chronic otitis, meningitis, and respiratory infections, a complete factor I deficiency was recognized . One of the patients died after fulminant sepsis . Hemolytic activity of the alternative pathway was not detectable in the patients' sera due to decreased plasma concentrations of C3, factor B and properdin . As a consequence of factor I deficiency, C3b could not be metabolized with the result that no C3-derived split products (C3dg/C3d) were detectable in the patients' sera . In vitro reconstitution with purified factor I restored the regulatory function in the patients' sera with the subsequent cleavage of C3b to C3c and C3dg . Factor H levels were decreased in all patients' sera and found to be tightly complexed with C3b resulting in a modified electrophoretic mobility . Upon factor I reconstitution, factor H was released from C3b regaining its beta 1 electrophoretic mobility . Complement-mediated biological functions like opsonization of bacteria, chemotactic activity and phagocytosis in these patients were impaired . The parents (cousins, 2nd degree) and 3/4 siblings had significantly reduced factor I plasma levels without further alteration in their complement profile . 3 of these obviously heterozygously deficient family members suffered from recurrent bacterial infections of different frequency and severity. J Prosthet Dent, 1998 Jan, 79(1), 79 - 89 The soft tissue response to osseointegrated dental implants; Weber HP et al.; The use of dental implants in the treatment of fully edentulous patients has become an important addition in oral/dental rehabilitation . The fact that these implants penetrate the oral mucosa can lead to the assumption that peri-implant tissues, similar to the periodontal tissues, are fulfilling an important function as a barrier to protect the bony anchorage underneath . It has been shown that insufficient plaque removal may lead to peri-implant tissue disease with bone loss similar to teeth . However, it is unclear how important this cause is as a source of implant failure compared with other factors, such as inadequate bone healing, unfavorable quantity and quality of bone, or (bio)mechanical and functional problems . It is also not understood if peri-implant epithelium and connective tissue are equally needed and/or qualified to slow down or prevent tissue breakdown as their periodontal counterparts . The scientific work focusing on peri-implant soft tissues has dramatically increased in the past few years . Most studies to date have examined and described their structure but little data exist on their true biologic function . This review analyzes the current understanding of morphologic and clinical features of the peri-implant soft tissues . Furthermore, evidence shall be provided that peri-implant soft tissues do not interfere with the current favorable results obtained when treating the edentulous patient with osseointegrated implants. Gynecol Obstet Invest, 1998, 45(1), 35 - 40 Stimulated polymorphonuclear leukocytes in vaginal secretions from patients with preterm labor; Matsubara S et al.; The purpose of this study was to examine evidence for the presence of activated vaginal leukocytes in women with preterm labor . Vaginal polymorphonuclear leukocytes from 7 patients in preterm labor (24-32 weeks of gestation) as well as from 7 control women with uncomplicated pregnancy were analyzed morphologically using transmission electron microscopy . Peroxidase and NADPH oxidase cytochemistry was also performed . Viable leukocytes were abundant in patients in preterm labor . Phagosomes, phagocytosis of bacteria, attachment of primary granules to the phagosomal membrane, and cell surface projections were observed in the vaginal leukocytes but not in the peripheral blood leukocytes . Peroxidase activity was visible on the cell surface, the phagosomal membrane, and the primary granules . NADPH oxidase activity was demonstrated on the cell surface of leukocytes . Morphological and cytochemical features indicated that vaginal polymorphonuclear leukocytes were stimulated in situ . Such stimulated leukocytes may play a role in the pathogenesis or pathophysiology of preterm labor. Clin Exp Immunol, 1998 Jan, 111(1), 36 - 47 The surface epithelium of recurrent infected palatine tonsils is rich in gammadelta T cells; Olofsson K et al.; Using a large panel of MoAbs in quantitative morphometric analysis of immunohistochemically stained tissue sections, we compared the frequency and distribution of immune cells in palatine tonsils from patients with recurrent tonsillitis (RT) and patients with idiopathic tonsillar hypertrophy (ITH) . We found that differences between the two patient groups in leucocyte populations were limited to the surface epithelium, whereas the cellular composition of interfollicular and follicular areas was similar . Most intraepithelial lymphocytes were CD8+ T cells in both groups . However, the number of intraepithelial T cells was significantly higher in RT compared with ITH . This was due to a selective increase in the number of intraepithelial CD8+ gammadelta T cells utilizing Vdelta1 and Vgamma9 . In both patient groups the majority of the intraepithelial gammadelta T cells expressed Vdelta1 and Vgamma9 . Subepithelially, gammadelta T cells utilizing Vgamma9 dominated over cells utilizing Vgamma8, while equal proportions expressed Vdelta1 and Vdelta2 . These results suggest that cells utilizing the otherwise rare combination Vdelta1/Vgamma9 in their T cell receptors (TCR) may constitute a major gammadelta T cell population in palatine tonsils and are probably reactive to antigens specific to the tonsillar milieu . Furthermore, they indicate that preferentially this gammadelta T cell subpopulation is involved in immune reactions within the surface epithelium in RT . We speculate that gammadelta T cells are involved in clearing infectious bacteria at the tonsillar surface and in limiting inflammatory responses in the tonsils . Both local expansion and infiltration of blood cells probably contribute to the high numbers of gammadelta T cells in RT patients. Appl Environ Microbiol, 1997 Dec, 63(12), 4853 - 8 Factors affecting lactate and malate utilization by Selenomonas ruminantium; Evans JD et al.; Lactate utilization by Selenomonas ruminantium is stimulated in the presence of malate . Because little information is available describing lactate-plus-malate utilization by this organism, the objective of this study was to evaluate factors affecting utilization of these two organic acids by two strains of S . ruminantium . When S . ruminantium HD4 and H18 were grown in batch culture on DL-lactate and DL-malate, both strains coutilized both organic acids for the initial 20 to 24 h of incubation and acetate, propionate, and succinate accumulated . However, when malate and succinate concentrations reached 7 mM, malate utilization ceased, and with strain H18, there was a complete cessation of DL-lactate utilization . Malate utilization by both strains was also inhibited in the presence of glucose . S . ruminantium HD4 was unable to grow on 6 mM DL-lactate at extracellular pH 5.5 in continuous culture (dilution rate, 0.05 h-1) and washed out of the culture vessel . Addition of 8 mM DL-malate to the medium prevented washout on 6 mM DL-lactate at pH 5.5 and resulted in succinate accumulation . Addition of malate also increased bacterial protein, acetate, and propionate concentrations in continuous culture . These results suggest that 8 mM DL-malate enhances the ability of strain HD4 to grow on 6 mM DL-lactate at extracellular pH 5.5. Ann Acad Med Stetin, 1997, 43, 143 - 59 {Diagnostic value of different methods applied for detecting Helicobacter pylori infection in gastric mucosa}; Korzonek M; The aim of the paper comprised: 1) estimating the incidence of Helicobacter pylori (H . pylori) infection in subjects directed to undergo endoscopic examination due to ailments involving the upper segments of the alimentary tract, 2) determination of the degree of H . pylorii infection detectability on the basis of invasive methods (bacterial culture, urease test and histological examination of specimens stained by Giemsa method) and non-invasive (serological investigation, skin test) with endoscopic image and histopathologic changes in gastric mucosa being talken into consideration, 3) assessing the titer of class IgG anti-H . pylori antibodies in subjects with endoscopic and histopathologic changes of gastric mucosa, infected by H . pylori, as well as persons after eradication of bacteria, 4) estimating the diagnostic value of individual methods . The study material consisted of 428 patients (224 women and 204 men) investigated at Endoscopy Laboratory of Internal Diseases Clinic in the years 1991-1994 . Bacterial cultures and urease tests were performed in all the studied subjects, 230 specimens stained by Giemsa method were examined histopathologically, in 351 subjects the titer of class IgG anti-H . pylori antibodies in blood serum were determined by ELISA method, while in 73 persons the skin test from the suspension of dead bacterial colonies was carried out . Histopathological examination of mucosal sections was accomplished in 396 studied subjects, assessing the histological type of mucosal inflammation in H . pylorii infected persons and in those having not been infected . The applicability of the respective diagnostic methods was examined by calculating the sensitivity and specificity of the given method, and taking into account the results of bacterial culture as the reference method . It is evident from the performed investigations that the majority of subjects, having been endoscopically studied because of ailments stemming from the upper segment of the alimentary tract, were infected by H . pylori (Tab . 1, 2, 3) . H . pylori colonisation in gastric mucosa was often accompanied by pathological changes of the stomach and duodenum in the endoscopic and histopathologic images . In the routine clinical diagnostics the urease test, histological examination of specimens stained by Giemsa method as well as bacterial culture are valuable methods of detecting H . pylorii infection . With regard to medium and high values of the titer of antibodies IgG anti-H . pylori, the serological investigation as a non-invasive method displays a high compliance with positive results of invasive examinations (Tab . 5, Fig . 1, 2, 3) . An early skin test may become a screening method for detecting H . pylori infection, after its methodical modification. Rozhl Chir, 1997 Oct, 76(10), 510 - 3 {Infections in biliary surgery}; Krikava K et al.; The authors selected among patients who were operated in 1992-1996 on account of affections of the gallbladder and biliary pathways those patients where aerobic and anaerobic bile cultivation was made . The results of the detected bacterial species and their sensitivity to ATB are given in tables . According to the author these investigations may be of benefit to the surgeon when he has to decide on the administration of ARTB without knowing the bacterial strain (prevention or therapy) and thus overcome the time when he will know the results of cultivation. Ann Ist Super Sanita, 1997, 33(2), 225 - 9 Molecular characterisation of Lyme disease borreliae using RAPD analysis and 16S rDNA sequencing; Bandi C et al.; Here we report the use of random amplified polymorphic DNAs (RAPDs) and sequence analysis of the genes encoding for the small subunit ribosomal RNA (16S rDNA) for the characterisation of Borrelia burgdorferi sensu lato strains recovered from Ixodes ricinus and from Lyme disease patients . All strains examined were assigned to the species Borrelia garinii . However, both RAPDs and 16S rDNAs revealed a level of genetic variation among the strains which appears higher than expected for a bacterial species . In addition, the data obtained agree with the clonal theory applied to Borrelia burgdorferi s.l . for explaining some traits of its epidemiology . According to this theory, particular strains should spread rapidly, leading to the diffusion of bacteria with a particular chromosomal genotype . Our results reveal high genetic variation even among strains isolated in the same period from a restricted geographic area . Moreover, the data here reported indicate that clonal diffusion of antigenic characteristics could also occur. Proc Natl Acad Sci U S A, 1998 Jan 6, 95(1), 224 - 8 Universally conserved translation initiation factors; Kyrpides NC et al.; The process by which translation is initiated has long been considered similar in Bacteria and Eukarya but accomplished by a different unrelated set of factors in the two cases . This not only implies separate evolutionary histories for the two but also implies that at the universal ancestor stage, a translation initiation mechanism either did not exist or was of a different nature than the extant processes . We demonstrate herein that (i) the "analogous" translation initiation factors IF-1 and eIF-1A are actually related in sequence, (ii) the "eukaryotic" translation factor SUI1 is universal in distribution, and (iii) the eukaryotic/archaeal translation factor eIF-5A is homologous to the bacterial translation factor EF-P . Thus, the rudiments of translation initiation would seem to have been present in the universal ancestor stage . However, significant development and refinement subsequently occurred independently on both the bacterial lineage and on the archaeal/eukaryotic line. J Mol Evol, 1997 Dec, 45(6), 708 - 11 ThiD-TenA: a gene pair fusion in eukaryotes; Ouzounis CA et al.; Computational analysis of the hypothetical open reading frame MJ0236 from Methanococcus jannaschii reveals its membership to a family of bacterial and eukaryotic proteins, predicted to be the HMP-P kinases involved in thiamin biosyntheis (ThiD) . The eukaryotic members of this family contain a C-terminal extension similar to a bacterial transcriptional activator (TenA), thus pointing to a fusion event that took place during cellular evolution . The C-terminal domain is absent from M . jannaschii . The significance of this observation is two-fold: first, this is a case where a fusion protein contains two domains with an unusual phylogenetic distribution, and second, the TenA domain is a rare case of a gene family involved in transcription present both in bacteria and eukaryotes. Nucleic Acids Res, 1998 Jan 1, 26(1), 351 - 2 The ribonuclease P database; Brown JW; Ribonuclease P is responsible for the 5'-maturation of tRNA precursors . Ribonuclease P is a ribonucleoprotein, and in bacteria the RNA subunit alone is catalytically active in vitro , i.e., it is a ribozyme . The Ribonuclease P Database is a compilation of ribonuclease P sequences, sequence alignments, secondary structures, three-dimensional models, and accessory information, available via the World Wide Web . Poult Sci, 1998 Jan, 77(1), 41 - 6 Peanut hulls as a litter source for broiler breeder replacement pullets; Lien RJ et al.; Broiler breeder pullets were reared on either peanut hulls or pine shavings to determine effects of litter type on growth performance and litter characteristics . Pullets were reared to 20 wk of age in rooms initially bedded with 8 cm of clean shavings or hulls . Heating and ventilation were standardized in all rooms . Restricted skip-a-day feeding was used to attain recommended growth curves . Water was continuously provided for ad libitum consumption . Litter and environmental variables were measured throughout rearing and 2 wk after pullets were removed from the litter materials . Feed consumption, BW, mortality, and uniformity at 20 wk were not affected by litter type; however, gizzard weights were decreased in pullets reared on hulls . Litter bulk density increased with use and was greater for shavings through 11 wk, but not thereafter . Particle size decreased with use in both litter types . Through 11 wk, there were more particles in the > 4 mm range and less in the < 1.7 mm range with hulls . Litter moisture increased with use but was not affected by litter type . Litter pH was greater in unused shavings, but during and after use was generally greater in hulls . With both litter types, litter and environmental ammonia levels increased to 11 wk then decreased; however, this effect was more pronounced for hulls . Bacteria populations were not affected by litter type; however, greater fungal populations were observed in shavings at 7 and 15 wk . Aflatoxins were detected in unused hulls but not shavings . Because aflatoxin levels decreased during use and Aspergillus flavus and Aspergillus parasiticus populations were not detected in samples collected during use, aflatoxins observed were presumed to have been formed prior to use . Peanut hulls performed similarly to pine shavings as a litter source for breeder pullets; however, the specific influence of the aflatoxins contained in this litter source on bird performance deserves further study. J Immunol, 1998 Feb 15, 160(4), 1886 - 93 Morphine enhances macrophage apoptosis; Singhal PC et al.; Laboratory data indicate that morphine decreases the numbier of peritoneal and alveolar macrophages (Mphi) and compromises their phagocytic capability for immune complexes and bacteria . We hypothesize that morphine decreases the number of, as well as compromises the phagocytic capability of, Mphi by programming their death . We studied the effect of morphine on Mphi apoptosis in vivo as well as in vitro . Peritoneal Mphi harvested from morphine-treated rats showed DNA fragmentation . Morphine enhanced murine Mphi (J 774.16) apoptosis in a dose-dependent manner . Human monocytes treated with morphine showed a classic ladder pattern in gel electrophoretic and end-labeling studies . Morphine promoted nitric oxide (NO) production both under basal and LPS-activated states . N(G)-nitro-L-arginine methyl ester (L-NAME) and N(G)-monomethyl-L-arginine monoacetate (L-NMMA), inhibitors of NO synthase, attenuated the morphine-induced generation of NO by Mphi . Morphine also enhanced Mphi mRNA expression of inducible NO synthase (iNOS) . Since morphine-induced Mphi apoptosis was inhibited by L-NAME and L-NMMA, it appears that morphine-induced Mphi apoptosis may be mediated through the generation of NO . Morphine promoted the synthesis of Bax and p53 proteins by Mphi . Moreover, IL-converting enzyme (ICE)-1 inhibitor attenuated morphine-induced Mphi apoptosis . These studies suggest that morphine activates the induction phase of the apoptotic pathway through accumulation of p53 . The effector phase of morphine-induced apoptosis appears to proceed through the accumulation of Bax and activation of ICE-1 . The present study provides a basis for a hypothesis that morphine may be directly compromising immune function by promoting Mphi apoptosis in patients with opiate addiction. J Immunol, 1998 Feb 15, 160(4), 1824 - 30 Toxoplasma gondii-infected cells are resistant to multiple inducers of apoptosis; Nash PB et al.; Infection with certain intracellular pathogens, including viruses and bacteria, may induce host cell apoptosis . On the other hand, infection with some viruses inhibits apoptosis . Complex protozoan parasites, including Toxoplasma gondii and members of Plasmodium, Leishmania, and Microsporidia, are also obligate intracellular pathogens, yet relatively little is known regarding their subversion of host cell functions . We now report that cells infected with T . gondii are resistant to multiple inducers of apoptosis, including Fas-dependent and Fas-independent CTL-mediated cytotoxicity, IL-2 deprivation, gamma irradiation, UV irradiation, and the calcium ionophore beauvericin . Inhibition of such a broad array of apoptosis inducers suggests that a mechanism common to many, or perhaps all, apoptotic pathways is involved . The inhibitory activity requires live intracellular parasite and ongoing protein synthesis . Despite T . gondii-mediated inhibition of DNA fragmentation, infected cells can still be lysed by CTL. Clin Chim Acta, 1997 Nov 28, 267(2), 183 - 96 Detection of spiral and coccoid forms of Helicobacter pylori using a murine monoclonal antibody; Cao J et al.; Helicobacter pylori is the major cause of gastritis . The aim of this investigation was to develop a specific antibody, which recognizes both coccoid and spiral forms of Helicobacter pylori and to test this antibody on gastric biopsy sections known to harbour coccoid bacteria . Murine monoclonal antibodies against glycine-acid extracts of five strains of Helicobacter pylori were raised . Immunofluorescence and immunoelectron microscopy showed that one antibody of the IgG1 subclass was specific for both the spiral and coccoid forms . It reacted with a 28 kDa protein that was present in all the five strains tested . Using this antibody in an indirect immunofluorescence assay of formalin-fixed antral and corpus biopsy specimens from Helicobacter pylori-associated gastritis patients showed that nine of the nine antral and five of six corpus specimens harboured the coccoid form of Helicobacter pylori . This technique thus provides a rapid and specific detection of both the spiral and coccoid forms. Digestion, 1998, 59(1), 1 - 15 A century of Helicobacter pylori: paradigms lost-paradigms regained; Kidd M et al.; The investigation of gastric bacteria properly began in the latter half of the nineteenth century when microscope resolution had sufficiently advanced . Whilst a bacterial etiology was demonstrated for dysentery, tuberculosis and syphilitic ulcers, problems in the isolation and culture of pure strains circumvented a role for bacteria in gastric pathology . Furthermore, dogma and the intellectual chorus were in harmony advocating that gastric acid was critical in ulcer disease . The consideration of a role for a pathogen or pepsin was regarded as whimsical in the context of mucosal ulceration . Indeed, the effects of acid inhibitory agents were held as gospel truth whilst the use of antibiotics or metallic ions were deemed to be quackery or at least ill judged . Nonetheless, spiral-shaped bacteria had been identified in both mucosa and gastric contents of patients as early as 1889 . Elegant studies had documented the infectivity of these organisms, and suggested but not proven a causative role in gastric disease . The prescient identification by Doenges of organisms associated with gastritis in both man and monkey, was buried by the observations of Palmer, and an opportunity for early progress lost . It required two decades and Antipodean pathological perspicacity to elucidate the warren of previous archaic gastric bacterial misinformation . The subsequent marshalling of clinical and pathological data established the fatal flaw in the mucosa to be bacteria and not only acid on the mucus shore . It is now widely apparent that Helicobacter is ubiquitous, pathological and, a century after its initial discovery, still remains a paradox of as yet incompletely determined biological consequence . It is of note that an organic helical configuration has twice in this century provided biological information of unique import. Biochim Biophys Acta, 1998 Jan 8, 1379(1), 69 - 75 Intestinal flora is not an intermediate in the phylloquinone-menaquinone-4 conversion in the rat; Ronden JE et al.; To elucidate the role of intestinal bacteria in the conversion of phylloquinone into menaquinone-4 (MK-4) we investigated the tissue distribution of vitamin K in germ-free rats . The rats were made vitamin K deficient by feeding a vitamin K-free diet for 13 days . In a subsequent period of 6 days, phylloquinone and menadione were supplied via the drinking water in concentrations of 10 and 50 micromol l(-1) . Menadione supplementation led to high levels of tissue MK-4, particularly in extrahepatic tissues like pancreas, aorta, fat and brain . Liver and serum were low in MK-4 . Phylloquinone supplementation resulted in higher phylloquinone levels in all tissues when compared with vitamin K-deficient values . The main target organs were liver, heart and fat . Remarkably, tissue MK-4 levels were also higher after the phylloquinone supplementation . The MK-4 tissue distribution pattern after phylloquinone intake was comparable with that found after menadione intake . Our results demonstrate that the conversion of phylloquinone into MK-4 in extrahepatic tissues may occur in the absence of an intestinal bacterial population and is tissue specific . A specific function for extrahepatic MK-4 or a reason for this biochemical conversion of phylloquinone into MK-4 remains unclear thus far. Crit Care Med, 1998 Feb, 26(2), 392 - 408 Practice parameters for evaluating new fever in critically ill adult patients . Task Force of the American College of Critical Care Medicine of the Society of Critical Care Medicine in collaboration with the Infectious Disease Society of America; O'Grady NP et al.; OBJECTIVE: To develop practice parameters for the evaluation of adult patients who develop a new fever in the intensive care unit (ICU) for the purpose of guiding clinical practice . PARTICIPANTS: A task force of 13 experts in disciplines related to critical care medicine, infectious diseases, and surgery was convened from the membership of the Society of Critical Care Medicine, and the Infectious Disease Society of America . EVIDENCE: The task force members provided the personal experience and determined the published literature (MEDLINE articles, textbooks, etc.) from which consensus would be sought . Published literature was reviewed and classified into one of four categories, according to study design and scientific value . CONSENSUS PROCESS: The task force met several times in person and twice monthly by teleconference over a 1-yr period of time to identify the pertinent literature and arrive at consensus recommendations . Consideration was given to the relationship between the weight of scientific evidence and the experts' opinions . Draft documents were composed and debated by the task force until consensus was reached by nominal group process . CONCLUSIONS: The panel concluded that, because fever can have many infectious and noninfectious etiologies, a new fever in a patient in the ICU should trigger a careful clinical assessment rather than automatic orders for laboratory and radiologic tests . A cost-conscious approach to obtaining cultures and imaging studies should be undertaken if it is indicated after a clinical evaluation . The goal of such an approach is to determine, in a directed manner, whether or not infection is present, so additional testing can be avoided and therapeutic options can be made. Microbiology, 1998 Jan, 144 ( Pt 1), 229 - 39 Metabolism is required for chemotaxis to sugars in Rhodobacter sphaeroides; Jeziore-Sassoon Y et al.; Chemotaxis towards carbohydrates is mediated, in enteric bacteria, either by the transport-independent, methylation-dependent chemotaxis pathway or by transport and phosphorylation via the phosphoenolpyruvate (PEP)-dependent phosphotransferase system (PTS) . This study shows that Rhodobacter sphaeroides is chemotactic to a range of carbohydrates but the response involves neither the classical methyl-accepting chemotaxis protein (MCP) pathway nor the PTS transport pathway . The chemoattractant fructose was transported by a fructose-specific PTS system, but transport through this system did not appear to cause a chemotactic signal . Chemotaxis to sugars was inducible and occurred with the induction of carbohydrate transport systems and with substrate incorporation . A mutation of the glucose-6-phosphate dehydrogenase gene (zwf) inhibited chemotaxis towards substrates metabolized by this pathway although transport was unaffected . Chemotaxis to other, unrelated, chemoattractants (e.g . succinate) was unaffected . These data, in conjunction with the fact that mannitol and fructose (which utilize different transport pathways) compete in chemotaxis assays, suggest that in R . sphaeroides the chemotactic signal is likely to be generated by metabolic intermediates or the activities of the electron-transport chain and not by a cell-surface receptor or the rate or mode of substrate transport. Microbiology, 1998 Jan, 144 ( Pt 1), 183 - 91 pqqA is not required for biosynthesis of pyrroloquinoline quinone in Methylobacterium extorquens AM1; Toyama H et al.; Methylobacterium extorquens AM1 is a facultative methylotroph that oxidizes methanol via the pyrroloquinoline quinone (PQQ)-linked enzyme methanol dehydrogenase . In M . extorquens AM1 and other PQQ-synthesizing bacteria, several genes are involved in the synthesis of PQQ and one of these, pqqA, has been proposed to encode a peptide precursor of PQQ . In other PQQ-synthesizing bacteria, pqqA is required for PQQ production . In this study, it is shown that both deletion and insertion mutants of pqqA in M . extorquens AM1 grow normally on methanol and produce PQQ . The level of PQQ production is reduced in the insertion mutant, but it is sufficient to allow normal growth on methanol . These results suggest either that a different peptide in M . extorquens AM1 can substitute for PqqA in pqqA mutants, or that PqqA-like peptides may not be obligatory precursors of PQQ . In addition, it is shown that the methanol oxidation transcriptional regulator gene, mxbM, is required for normal methanol induction of PQQ synthesis. Int J Immunopharmacol, 1997 Jun, 19(6), 355 - 8 Potent anti-inflammatory activity of pheophytin a derived from edible green alga, Enteromorpha prolifera (Sujiao-nori); Okai Y et al.; Recently, a chlorophyll-related compound, pheophytin a, has been identified from an edible green alga, Enteromorpha prolifera (Sujiao-nori in Japanese) as a potent suppressive substance against genotoxin-induced umu C gene expression in a tester bacteria (Okai and Higashi-Okai, 1997, J . Sci . Food Agricul . 71, 531-535) . In the present study, anti-inflammatory effects of pheophytin a from Enteromorpha prolifera have been analyzed using in vitro and in vivo experiments . 1 . Pheophytin a suppressed the production of superoxide anion (O2-) in mouse macrophages induced by 12-O-tetradecanoylphorbol-13-acetate (TPA) using the cytochrome C reduction method . 2 . Pheophytin a caused a suppressive effect against formyl-Met-Leu-Phe, (FMLP)-induced chemotaxis of human polymorphonuclear leukocytes (PMNs) in Boyden's chamber experiment . 3 . Pheophytin a exhibited a significant suppression against TPA-induced inflammatory reaction such as edema formation in BALB/c mouse ear . These results suggest that pheophytin a from Enteromorpha prolifera has a potent anti-inflammatory activity. Oral Microbiol Immunol, 1997 Oct, 12(5), 303 - 10 Estimation of serum antibody to subgingival species using checkerboard immunoblotting; Sakellari D et al.; Measurement of serum antibody to subgingival bacterial species has been useful in discriminating possible periodontal pathogens and in assessing the host's immune response to subgingival species . An immunoassay system was developed to measure the level of serum antibody to multiple subgingival species in multiple serum samples on a single nitrocellulose membrane . The principle steps of the assay are the following: 1) binding of antigens from bacterial preparations and protein A in parallel lanes on nitrocellulose membranes; 2) incubation of known concentrations of human immunoglobulin as well as various dilutions of serum from patients in lanes perpendicular to the antigen lanes; 3) incubation of the membrane with a peroxidase-conjugated second antibody; 4) detection of positive reactions by enhanced chemiluminescence . Emitted light was captured on a photographic film in which the positive reactions appeared as squares at the intersections of antigens with appropriate antibody . Antibody was quantified using a laser densitometer to compare the signal intensity of unknown samples with the ones generated by known amounts of human immunoglobulin captured on the same membrane . The assay permitted simultaneous screening and/or quantification of antibody in as many as 45 serum samples against up to 45 bacterial species . The mean and standard error of the coefficients of variation for replicates within an assay averaged 7.3 +/- 2.3% . Coefficients of variation of the assay run on different days for serum antibody to a range of subgingival species averaged 10.1 +/- 2.1% . Checkerboard immunoblotting is a simple and rapid immunoassay to permit quantification and/or screening of antibody to multiple subgingival species or antigens in multiple serum samples. Oral Microbiol Immunol, 1996 Dec, 11(6), 402 - 6 Adhesion of Porphyromonas gingivalis fimbriae to human gingival cell line Ca9-22; Hirose K et al.; In this study, we examined the effects of selected environmental factors on the adhesion of Porphyromonas gingivalis fimbriae, an important structure involved in attachment of the bacteria to human gingival cells . The human gingival carcinoma cell line Ca9-22 was grown in microculture plates, and adherence was detected by use of 125I-labeled fimbriae . Adhesion was increased by changes in pH from 7.0-8.0, but was decreased by increase in the sodium chloride concentration above 0.15 M . Trypsin treatment of Ca9-22 cells also augmented adhesion of the fimbriae to the cells . These results indicate that fimbrial adhesion to gingival cells is controlled by various environmental factors, and the data on trypsin treatment suggest that elevated levels of protease in the gingival sulcus, such as can occur with poor oral hygiene and gingivitis, may expose adhesion molecules on the gingival cell surface, thereby promoting the attachment of P . gingivalis fimbriae. Oral Dis, 1997 Sep, 3(3), 167 - 71 Priming response to inflammatory mediators in hyperreactive peripheral neutrophils from adult periodontitis; Gustafsson A et al.; OBJECTIVES: To study the response to in vitro priming of peripheral neutrophils from patients with periodontitis compared to healthy controls . Peripheral neutrophils from these patients had shown increased production of oxygen radicals after activation with opsonized bacteria and a difference in priming response could suggest an explanation for this hyperreactivity . MATERIALS AND METHODS: Peripheral neutrophils from a group of patients with periodontitis and from age- and sex-matched controls were preincubated with tumor necrosis factor alpha, lipopolysaccharide (LPS), formyl-methionyl-leucyl-phenylalanine and the tetra peptide arginyl-glycyl-aspartate-serine . After preincubation, the cells were activated with gammaglobulin opsonized-bacteria, i.e., a Fc gamma R-stimulation . The priming effect was assessed as the production of oxygen radicals and as the degranulation or primary granules . RESULTS: Showed that the patients had a slightly lower responsiveness to priming than had the controls . This difference in priming response was most pronounced when measured as degranulation of primary granules after preincubation with LPS and 20 min of activation . CONCLUSIONS: This study shows no difference in response to priming, with optimal concentrations of inflammatory mediators, between peripheral neutrophils from patients with adult periodontitis and healthy controls. Ann Univ Mariae Curie Sklodowska {Med}, 1996, 51, 195 - 202 {Reactions of 1-(X-benzoyl)-4-R-thiosemicarbazide with chloroacetone and omega-bromoacetophenone . VI . 4-(o-tolyl)-thiosemicarbazide of o-nitro - and p-nitrobenzoic acid}; Bielak E et al.; The reactions of 4-(o-tolyl)-thiosemicarbazide of o-nitro- and p-nitrobenzoic acid (Ik, l) with chloroacetone and omega-bromoacetophenone were investigated in: methanolic medium (method D); methanolic medium in the presence of anhydrous CH3COONa (method E); methanolic medium in the presence of N(C2H5)3 (method F) . The properties of compounds IIu-y and IIIu-y were determined under the conditions of basic and acidic hydrolysis . The results of UV, IR and NMR spectroscopic measurements were reported. J Mol Biol, 1998 Jan 16, 275(2), 337 - 46 Crystal structure of the cyanobacterial metallothionein repressor SmtB: a model for metalloregulatory proteins; Cook WJ et al.; SmtB from Synechococcus PCC7942 is a trans-acting dimeric repressor that is required for Zn(2+)-responsive expression of the metallothionein SmtA . The structure of SmtB was solved using multiple isomorphous replacement techniques and refined at 2.2 A resolution by simulated annealing to an R-factor of 0.218 . SmtB displays the classical helix-turn-helix motif found in many DNA-binding proteins . It has an alpha + beta topology, and the arrangement of the three core helices and the beta hairpin is similar to the HNF-3/fork head, CAP and diphtheria toxin repressor proteins . Although there is no zinc in the crystal structure, analysis of a mercuric acetate derivative suggests a total of four Zn2+ binding sites in the dimer . Two of these putative sites are at the opposite ends of the dimer, while the other two are at the dimer interface and are formed by residues contributed from each monomer . The structure of the dimer is such that simultaneous binding for both recognition helices to DNA would require either a bend in the DNA helix or a conformational change in the dimer . The structure of Synechococcus SmtB is the first in this family of metal-binding DNA repressors. J Clin Microbiol, 1998 Feb, 36(2), 585 - 6 Quality control limits for dilution and disk diffusion susceptibility tests of trovafloxacin against eight quality control strains; Fuchs PC et al.; A 10-laboratory collaborative effort was designed to generate data to propose quality control limits for susceptibility tests of trovafloxacin . Broth microdilution, agar dilution, and disk diffusion tests were evaluated with eight different control strains . All tests were reproducible, and control limits are proposed. J Clin Microbiol, 1998 Feb, 36(2), 443 - 8 Improved diagnostic PCR assay for Actinobacillus pleuropneumoniae based on the nucleotide sequence of an outer membrane lipoprotein; Gram T et al.; The gene (omlA) coding for an outer membrane protein of Actinobacillus pleuropneumoniae serotypes 1 and 5 has been described earlier and has formed the basis for development of a specific PCR assay . The corresponding regions of all 12 A . pleuropneumoniae reference strains of biovar 1 were sequenced . Alignment of the sequences revealed conserved terminal and variable middle regions, which divided the reference strains into four distinct groups . Primers were selected from the conserved 5' and 3' termini of the gene . A 950-bp amplicon was obtained from each of 102 tested field isolates of A . pleuropneumoniae obtained from lungs . Their identity was verified by sequencing approximately 500 bp of the amplification product from 50 of the A . pleuropneumoniae isolates, which all showed the expected DNA sequence characteristic of the serotype . To test the specificity of the reaction, 23 other bacterial species related to A . pleuropneumoniae or isolated from pigs were assayed . They were all found negative in the PCR, as were tonsil cultures from 50 pigs of an A . pleuropneumoniae-negative herd . The sensitivity assessed by agarose gel analysis of the PCR product was 10(2) CFU/PCR test tube . The specificity and sensitivity of this PCR compared to those of culture suggest the use of this PCR for routine identification of A . pleuropneumoniae. Eur J Clin Invest, 1997 Dec, 27(12), 1030 - 7 Oxidative autoaggression by phagocytes in human peritonitis; Billing AG et al.; In 60 abdominal exudates from patients with the diagnosis of either acute or persistent (chronic) peritonitis, indicators of phagocyte-derived oxidative systems (myeloperoxidase, chemiluminescence) and proteases (elastase) were measured . These exudates reveal a picture of maximal inflammatory activation . Both types of exudates (30 each) showed a significant influx of inflammatory cells, with the mean leucocyte count being 73,000 microL and 32,000 microL-1 respectively . Local myeloperoxidase concentrations were approximately 1000-fold greater than that of normal plasma . Spontaneous and elicitable chemiluminescence--indicators of phagocyte respiratory burst activity--were dramatically increased . In addition, levels of extracellularly released elastase (from neutrophils) were found to be up to about 1000-fold that of normal plasma values . Although most of the elastase detected in the exudates was complexed with alpha 1-proteinase inhibitor (alpha 1 PI), enzymatically active elastase could be measured in approximately 60% of the samples being investigated . As there was an excess of immunoreactive alpha 1 PI in these exudates, the free elastase activity implies that much of the alpha 1 PI was inactive, presumably subjected to oxidative destruction . Moreover, a trypsin-inhibitory activity to antigen ratio below 1 (mean = 0.81) in 75% of the purulent exudates indicated also partial proteolytic degradation of alpha 1 PI . In contrast, 16 clear exudates (no bacteria, white cell count below 500 microL-1) taken from the non-infected peritoneal cavity of patients undergoing intra-abdominal surgery revealed a similar permeability increase of the peritoneum but did not show relevant oxidative and proteolytic activity or destruction of alpha 1 PI compared with purulent specimens . Thus, only the inflammatory process of peritonitis appears to result in an overwhelming local phagocytic activity that initiates and maintains protease inhibitor consumption and/or inactivation . The tremendous oxidative potential found in purulent exudates may cause destruction in a wide variety of defence systems. Zentralbl Veterinarmed A, 1997 Dec, 44(9-10), 595 - 601 Hydrocephalus with periventricular encephalitis in the dog; Cantile C et al.; A 3-month-old female Dalmatian dog and a 2.5-month-old Poodle dog were referred with a sudden onset of neurological syndrome consistent with hydrocephalus . Clinical signs included depression, severe ataxia, eye abnormalities and skull enlargement in one case . Postmortem examination revealed severe internal hydrocephalus with cavitation of the cerebral white matter associated with necrotizing and inflammatory lesions of the periventricular nervous tissue . Although no bacteria were isolated from cerebrospinal fluid and no infectious agents were detected in the brains, an infectious etiology was postulated. Eur J Immunol, 1997 Dec, 27(12), 3461 - 70 Induction of interleukin-12 p40 transcript by CD40 ligation via activation of nuclear factor-kappaB; Yoshimoto T et al.; Interleukin-12 is produced in response to infection with bacteria or parasites or to bacterial constituents such as LPS in monocytes/macrophages and dendritic cells, and also generated by the interaction between activated T cells and antigen-presenting cells via CD40-CD40 ligand (CD40L) . So far, transcriptional analyses of p40 have been carried out only using bacterial constituents such as LPS as stimuli . In the present study, we have characterized the transcriptional induction of p40 by CD40 ligation in a human B lymphoblastoid cell line, Daudi, and a human acute monocytic leukemia cell line, THP-1 . These cells, stimulated by an agonistic monoclonal antibody against CD40 or by transfection with a CD40L expression vector, secreted p40 and showed enhanced p40 mRNA expression . Sequence analysis of the p40 promoter region identified two potential nuclear factor (NF)-kappaB binding sites conserved between mouse and human . Electrophoretic mobility shift assay revealed that the potential NF-kappaB binding sequence which is located around 120 bp upstream of the transcription initiation site in murine and human p40 genes formed an NF-kappaB complex with nuclear extract from Daudi cells stimulated by CD40 ligation . Moreover, transfection of Daudi cells with the polymerized NF-kappaB binding sequence ligated to a thymidine kinase/chloramphenicol acetyltransferase (CAT) reporter plasmid greatly induced CAT activity, but transfection with the polymerized mutated NF-kappaB binding sequence did not . These results suggest that the NF-kappaB binding site located around 120 bp upstream of the transcription initiation site in murine and human p40 promoter regions could be important for the p40 induction by CD40 ligation via activation of NF-kappaB. Appl Environ Microbiol, 1998 Feb, 64(2), 613 - 7 The use of fluorogenic substrates to measure fungal presence and activity in soil; Miller M et al.; Our objective was to determine if 4-methylumbelliferyl-labelled enzyme substrates could be used to detect and quantify specific components of chitinase and cellulase activities as specific indicators of the presence and activity of fungal biomass . The fluorogenic substrates 4-methylumbelliferyl (MUF) N-acetyl-beta-D-glucosaminide and MUF beta-D-lactoside were used for the detection and quantification of beta-N-acetylglucosaminidase (EC 3.2.1.30) (NAGase) and endo 1,4-beta-glucanase (EC 3.2.1.4)/cellobiohydrolase (EC 3.2.1.91) (CELase), respectively . Culture screenings on solid media showed a widespread ability to produce NAGase among a taxonomically diverse selection of fungi on media with and without added chitin . NAGase activity was expressed only in a limited number of bacteria and on media supplemented with chitin . The CELase activity was observed only in a limited number of fungi and bacteria . Bacterial CELase activity was expressed on agar media containing a cellulose-derived substrate . In soil samples, NAGase activity was significantly correlated with estimates of fungal biomass, based on the content of two fungus-specific indicator molecules, 18:2 omega 6 phospholipid fatty acid (PLFA) and ergosterol . CELase activity was significantly correlated with the PLFA-based estimate of fungal biomass in the soil, but no correlation was found with ergosterol-based estimates of fungal biomass. J Nat Prod, 1998 Jan, 61(1), 116 - 8 Purine and nucleoside metabolites from the Antarctic sponge Isodictya erinacea; Moon B et al.; The bright yellow sponge Isodictya erinacea is one of several chemically defended sponges found on the benthos of McMurdo Sound, Antarctica . An investigation of the metabolites from this sponge has resulted in the isolation of purine and nucleoside metabolites, including the previously unreported erinacean (1) and p-hydroxybenzaldehyde . The latter metabolite has been demonstrated to cause a feeding deterrence behavior in Perknaster fuscus, the major predator of antarctic sponges. Biochemistry (Mosc), 1997 Oct, 62(10), 1103 - 8 Ether lipids and platelet-activating factor: evolution and cellular function; Kulikov VI et al.; This review considers the relation between the evolution of ether lipids and platelet-activating factor (PAF) in living organisms for the first time . Ether lipids are shown to be the main structural lipid components in the cells of the most primitive organisms on the Earth; during evolution they were gradually substituted for lipids with ester and vinyl bonds . Synthesis of PAF has been found in some bacteria, protozoans, yeasts, plants, marine invertebrates, lower vertebrates, and mammals . The regulatory role of PAF is suggested to already appear in protozoans and later be maintained during the subsequent evolution of living organisms . During evolution, functions of PAF in the cell have been changing and enlarging, while ether lipids have been gradually losing their role as the main structural lipid component of the cells of living organisms. Gene, 1998 Jan 5, 206(1), 99 - 105 The mRNA for GST Pi from FRHK rhesus monkey kidney cells codes for an enzyme with activity towards 1-chloro-2,4-dinitrobenzene in spite of an I68F mutation; Swedmark S et al.; A cDNA library was constructed from mRNA of the rhesus monkey kidney cell line, FRHK, and the cDNA sequence for an FRHK glutathione S-transferase (GST) Pi was determined using a RACE method . This represents the first full-length monkey GST Pi sequence to be cloned and determined . The similarity to the human GST Pi was found to be extensive (more than 97%), the deduced protein differing only in six amino acids (aa) positions . FRHK GST Pi was expressed in bacteria and a recombinant protein was purified which demonstrated significant activity towards the substrates 1-chloro-2,4-dinitrobenzene (CDNB) and 1,2-epoxy-3-para-nitrophenoxypropane . Western blots also showed significant amounts of protein, both in the FRHK cells and transformed bacteria . The FRHK GST Pi was found to contain a phenylalanine at aa position 68, a position which is otherwise invariably occupied by an isoleucine in the GST Pi, Alpha, Mu and Beta class enzymes investigated . An isoleucine in this position is thus not essential for activity in the FRHK enzyme, unlike the human GST pi, where the exchange of Ile68 to a tyrosine (Manoharan, T.H, Gulick, A.M., Puchalski, R.B., Servais, A.L., Fahl, W.E., 1992 . J . Biol . Chem., 267, 18940-18945), resulted in total loss of activity . Phe68 was mutated to Ile in the FRHK GST Pi enzyme to determine whether the wild type amino acid conferred an impaired catalytic site . The resulting mutant did not show any changes in activity towards CDNB, clearly demonstrating that isoleucine at position 68 is not essential . Thus, the first monkey GST Pi enzyme has been characterized, an enzyme with many similarities to the human forms although it differs in an otherwise conserved residue at aa position 68 . This difference does not appear to affect the function of the FRHK GST Pi. Gene, 1997 Dec 31, 205(1-2), 103 - 7 CpG distribution patterns in methylated and non-methylated species; Shimizu TS et al.; To characterize the extent of DNA methylation and its possible biological roles in a wide variety of organisms, we have analyzed gene sequences extracted from the GenBank database . Sequences of both methylated and non-methylated species were used for comparative analysis . The local CpG dinucleotide distribution near the 5' ends of genes as well as the degree of overall CpG suppression/depletion in the entire gene region were examined in all complete gene sequences for each species . We show that the distribution patterns of CpG near the 5' region of genes differ among vertebrates, invertebrates, plants and bacteria . CpG island-like peaks in CpG O/E (observed/expected ratio) were observed not only in methylated species, but also in non-methylated species . In methylated non-vertebrates, overall CpG O/E values were lower, and peaks in the CpG profile of 5' regions were larger than in non-methylated species . We discuss the implications of such biases with respect to DNA methylation. Biol Chem, 1997 Dec, 378(12), 1413 - 9 The non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase: biochemistry, structure, occurrence and evolution; Habenicht A; The non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase catalyses the irreversible reaction of glyceraldehyde-3-phosphate to 3-phosphoglycerate by the reduction of NADP to NADPH . This is in contrast to the extensively analysed phosphorylating glyceraldehyde-3-phosphate dehydrogenases which catalyse the reversible reaction of glyceraldehyde-3-phosphate to 1,3-bisphosphoglycerate . Sequence analysis revealed that the non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase is not related to the phosphorylating glyceraldehyde-3-phosphate dehydrogenases but a member of the aldehyde dehydrogenase superfamily . The aldehyde dehydrogenases are of ancient origin and they have already existed in the progenote as indicated by phylogenetic analysis . Thus the non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase can be found in all three domains, archaea, bacteria and eukarya . The catalytic mechanism of the non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase and the other aldehyde dehydrogenases resembles a thioester mechanism involving the universally conserved cysteine 298 (pea GAPN) . The cofactor of the aldehyde dehydrogenases is bound in a new mode to a structure described as beta-alpha,beta-fold. Perspectives, 1997 Winter, 21(4), 15 - 6 Wound cleaning versus wound disinfection: a challenging dilemma; Phillips D et al.; Our experience with this resident has had a significant impact on our approach to wound management . We no longer accept normal saline as the only option for wound cleansing . Instead, we approach wound cleansing systematically using the wound cleansing model outlined in Figure 1 . If healing is not apparent, we critically review the situation and consider the bacterial status of the wound, the phase of wound healing, and the effects of wound cleansing versus wound disinfection on both the bacteria and the cells responsible for wound repair . In infected wounds or those colonized with a high bacterial count, careful attention is given to eradicating bacterial contamination . We carefully weigh the benefits of wound disinfection (eradication of bacterial contamination) against wound cleansing and the potential harm to the resident (delayed wound healing and unnecessary discomfort) . If the benefits of wound disinfection outweigh the potential harm to the resident, we choose wound disinfection and monitor its effectiveness on a daily basis . When bacterial contamination has been eradicated and the wound is clean, we resume wound cleansing with normal saline. Hum Reprod Update, 1997 Jul-Aug, 3(4), 301 - 10 Fertility regulating and immunotherapeutic vaccines reaching human trials stage; Talwar GP; The progress and current status of vaccines which induce antibodies against human chorionic gonadotrophin (HCG) and luteinizing hormone-releasing hormone (LHRH) are reviewed . Three vaccines devised against HCG have undergone phase I clinical trials documenting their safety, and reversibility . One of these, the heterospecies dimer (HSD)-HCG vaccine has also completed phase II efficacy trials in sexually active women of proven fertility . Immunization with the vaccine prevents pregnancy, as long as the antibody titres remain > or =50 ng/ml HCG bioneutralization capacity . There is no disturbance of menstrual regularity and women continue to ovulate normally . The antibody response is predominantly against an epitope in the core part of beta-HCG . Fertility is regained at titres <35 ng . These observations have laid the scientific foundations of a birth control vaccine . Research suggests the feasibility of making a cost-effective recombinant vaccine . The carriers tetanus toxoid (TT) and diptheria toxoid (DT) can be advantageously replaced by peptide determinants recognizing T, not B cells . In addition to optional fertility control, HCG vaccines may have tumour growth inhibition potential in lung cancers which produce HCG . The vaccine against LHRH can be used in both males and females . As it is a structurally conserved molecule, the same vaccine is applicable to both animals and humans . Antibodies against LHRH block the generation of gametes and sex steroids, with the result that the vaccine can be used for fertility control (domestic pets, prolongation of lactation amenorrhoea); as well as for sex hormone-dependent cancers . Phase I/phase II clinical trials have been conducted with the LHRH vaccine in advanced metastazing carcinoma of prostate patients with encouraging results . Bioeffective monoclonal antibodies have been developed against both LHRH and HCG . These can be 'humanized' and produced cost-effectively in bacteria and plants, thus paving the way for passive use of such antibodies for immunotherapy of cancers and fertility control. J Crit Care, 1997 Dec, 12(4), 193 - 9 Study of some phagocyte membrane receptors in patients receiving intravenous polyvalent immunoglobulins as adjunct treatment for nosocomial pneumonia; Martin C et al.; PURPOSE: Phagocytosis is a major mechanism of defense against bacterial infections . The ingestion of bacteria by phagocytes involves a variety of cell membrane recognition structures and, among them, immunoglobulin receptors . The aim of this study was to test the phagocytic activity of granulocytes and monocytes of intensive care unit (ICU) patients, and to evaluate the effects of intravenous polyvalent immunoglobulins (IVIG) used as adjunct treatment of nosocomial pneumonia on some phagocyte membrane receptors of these patients . MATERIALS AND METHODS: The phagocytic activity of granulocytes and monocytes of 41 mechanically ventilated patients with nosocomial bacterial pneumonia was studied during the acute phase of infection . These ICU patients were compared with 21 hospitalized, noninfected volunteer patients hospitalized in a medical ward . Peripheral blood granulocytes and monocytes were studied . Of the 41 ICU patients, after randomization, 21 received IVIG at a dose of 1 g/kg for 3 days . The 41 ICU patients were compared with the 21 non-ICU, noninfected hospitalized controls . The 21 ICU patients who received 3 days of IVIG were also compared with the 20 ICU patients not receiving IVIG . Cells were tested in standard immunoglobulin-free medium (fetal calf serum) and in the presence of patients' serum . Blood granulocytes and monocytes were purified and separately exposed to three types of particles: antibody-coated erythrocytes (to test immunoglobulin receptors), opsonized zymosan (to test C3 receptors), and glutaraldehyde-treated erythrocytes (to test lectinlike or other nonspecific binding sites) . Phagocytosis and superoxide anion production (oxidative burst) were measured . RESULTS: Granulocytes of ICU patients compared with those of non-ICU, noninfected patients exhibited a substantial decrease of zymosan ingestion (P < .05), whereas phagocytosis of other particles was normal . Monocytes from the ICU patients, compared with those of the non-ICU, noninfected patients, displayed an unselective overall decrease of phagocytic ability for the three particle types (P < .05) . The phagocytosis activity of the three membrane receptor species of blood monocytes and granulocytes of ICU patients was not significantly modified by the IVIG infusion . For both monocytes and granulocytes, no significant improvement was observed in the fraction of cells that ingested at least one foreign particle and the mean number of particles per cell having phagocytized at least one foreign particle . Granulocyte and monocyte functions were also tested by the production of reduced ferricytochrome and no significant improvement in the oxidative burst was observed after infusion of IVIG . CONCLUSION: Infected ICU patients display a deficiency of phagocytosis membrane receptors of blood granulocytes and monocytes . The addition of IVIG to standard therapy does not improve the phagocytic activity of ICU patients with nosocomial pneumonia. Z Gastroenterol, 1997 Oct, 35(10), 929 - 34 {Sclerosing cholangitis after burn injury}; Schmitt M et al.; We present the case of a 57-year-old woman who developed persistent jaundice and cholestasis four weeks after a severe burn injury (34% of the body surface, 19% second degree, 15% third degree) . In the endoscopic retrograde cholangiography four months later the characteristic picture of sclerosing cholangitis was found . A causal relation between the thermal injury and the development of sclerosing cholangitis is likely because of this coincidence and the absence of any preexisting disease, especially of the hepatobiliary system . Other cholestatic liver disease could be excluded . Possible pathogenetic mechanisms are discussed, especially the increased permeability of the gut mucosa after burn injuries with consecutive translocation of bacteria and toxins and their transport into the biliary tract. Int J Tuberc Lung Dis, 1997 Aug, 1(4), 370 - 6 Restriction fragment length polymorphism study of Mycobacterium tuberculosis in Thailand using IS6110 as probe; Palittapongarnpim P et al.; SETTING: Three referral hospitals in central Thailand . OBJECTIVE: To determine the population structure of Mycobacterium tuberculosis isolated from the referral hospitals . DESIGN: Study of 211 isolates of the bacteria received from the hospitals in central Thailand by Southern hybridization, with IS6110 probe and other probes when indicated . RESULTS: In 43 isolates only one copy of IS6110 was observed . These could be further differentiated by DR- and PGRS-specific probes . Two large groups of isolates with similar hybridization patterns were identified . The Beijing family, comprising 80 isolates, was previously reported to be commonly found in China, Mongolia, Thailand and Korea . The Nonthaburi group, comprising 29 isolates, were local strains . The age, sex and HIV status of the patients did not significantly correlate with the chance of being infected by isolates of any particular hybridization pattern . However, clustered isolates were found more commonly among the members of both the Beijing family and the Nonthaburi group . CONCLUSION: Southern hybridization with IS6110 was found to be useful in studying the epidemiology of tuberculosis in Thailand . The existence of the Beijing family was confirmed . The unusually wide spread of the Beijing family in several countries in Asia merits further investigation. Pediatr Res, 1998 Jan, 43(1), 84 - 90 Butyrate enhances interleukin (IL)-8 secretion by intestinal epithelial cells in response to IL-1beta and lipopolysaccharide; Fusunyan RD et al.; Intestinal epithelial (Caco-2) cells secrete the chemokine, IL-8, after stimulation with IL-1beta, but not after lipopolysaccharide . Butyrate is a short chain fatty acid derived from the metabolism of intestinal contents by gut bacteria . Butyrate concentrations reflect, therefore, the bacterial microenvironment established within the intestine . We hypothesized that butyrate may alter the secretion of IL-8 by intestinal epithelial cells in response to stimulation by IL-1beta or lipopolysaccharide . Caco-2 cells were incubated in varying concentrations of sodium butyrate (0-20 mM) for 24 h before stimulation with lipopolysaccharide or IL-1beta . IL-8 secretion was measured over 24 h by ELISA . IL-8 mRNA accumulation was detected by Northern blots . Lipopolysaccharide induced the secretion of IL-8 only after Caco-2 cells cells had been cultured with sodium butyrate . Furthermore, butyrate significantly enhanced IL-8 secretion by cells stimulated with IL-1beta . Butyrate also increased IL-8 mRNA accumulation in stimulated Caco-2 cells . Intestinal epithelial cells can, therefore, be primed by butyrate to become activated by lipopolysaccharide and proinflammatory cytokines . This may represent a mechanism by which intestinal epithelial cells can regulate intestinal inflammation in response to changes in the intestinal milieu. J Clin Microbiol, 1998 Jan, 36(1), 251 - 4 Development of an immunomagnetic method for selective isolation of Actinobacillus pleuropneumoniae serotype 1 from tonsils; Gagne A et al.; An immunomagnetic separation technique (IMS) for the selective isolation of Actinobacillus pleuropneumoniae serotype 1 was developed . Superparamagnetic polystyrene beads (immunomagnetic beads {IMBs}) were coated with purified rabbit immunoglobulin G specific for A . pleuropneumoniae serotype 1 . The antibody concentration, the number of IMBs, the incubation time, and the temperature of incubation influenced the recovery of the target bacteria . The sensitivity of the IMS technique was 1,000-fold higher than that of direct culture . When tonsils from animals from infected herds were tested, significantly more positive tonsils were detected by the IMS technique (68%) than by the standard procedures (22%) . The method represents an innovative and highly sensitive approach for the isolation of A . pleuropneumoniae from carrier animals. Heart Lung, 1997 Nov-Dec, 26(6), 419 - 29 Ventilator-associated pneumonia: clinical significance and implications for nursing; Grap MJ et al.; Pneumonia is the second most common nosocomial infection in the United States and the leading cause of death from nosocomial infections . Intubation and mechanical ventilation greatly increase the risk of bacterial pneumonia . Ventilator-associated pneumonia (VAP) occurs in a patient treated with mechanical ventilation, and it is neither present nor developing at the time of intubation; it is a serious problem--with significant morbidity and mortality rates . Aspiration of bacteria from the oropharynx, leakage of contaminated secretions around the endotracheal tube, patient position, and cross-contamination from respiratory equipment and health care providers are important factors in the development of VAP . Nurses caring for patients treated with mechanical ventilation must recognize risk factors and include strategies for reducing these factors as part of their nursing care . This article summarizes the literature related to VAP: its incidence, associated factors, diagnosis, and current therapies, with an emphasis on nursing implications in the care of these patients. J Reprod Immunol, 1997 Nov 30, 36(1-2), 93 - 109 Pre-term labor: an intra-uterine inflammatory response syndrome? Dudley DJ. Emerging concepts of sepsis suggest that the host response to an infectious stimulus results in some cases of uncontrolled release of inflammatory cytokines leading to signs of sepsis . Systemic inflammatory response syndrome (SIRS) has been suggested as a diagnosis when no etiologic organism can be found . Infection may account for up to 30% of cases of pre-term labor, and may either be clinically-evident or sub-clinical . Inflammatory cytokines can be detected in elevated concentrations in the amniotic fluid and plasma of women with pre-term labor, and human gestational tissues are potentially rich sources of inflammatory cytokines, as found in in vivo and in vitro studies . Also, maternal decidua and fetal membranes produce mRNA for inflammatory cytokines in the setting of infection-associated pre-term labor and normal term labor . Notably, anti-inflammatory cytokines, such as interleukin-10 (IL-10) do not appear to be present in substantial quantities in these pathophysiologic and physiologic conditions . Animal models indicate that pre-term labor can be stimulated by bacteria, bacterial cell wall products, and inflammatory cytokines such as IL-1 and tumor necrosis factor . These findings suggest that: (1) infectious stimuli may result in the liberation of inflammatory cytokines from gestational tissues leading inevitably to pre-term labor and delivery; (2) inhibition of this process may either be overcome or abrogated, and (3) the mechanisms regulating cytokine production in maternal and fetal tissues are disturbed . Thus, pre-term labor associated with sub-clinical infection may result in a dysregulated local inflammatory response, in which the maternal host response causes an 'intra-uterine inflammatory response syndrome' leading to pre-term labor and delivery. Methods, 1997 Oct, 13(2), 112 - 22 Tagging genes and trapping promoters in Toxoplasma gondii by insertional mutagenesis; Roos DS et al.; Plasmid vectors that incorporate sequence elements from the dehydrofolate reductase-thymidylate synthase (DHFR-TS) locus of Toxoplasma gondii integrate into the parasite genome with remarkably high frequency (>1% of transfected parasites) . These vectors may-but need not-include mutant DHFR-TS alleles that confer pyrimethamine resistance to transgenic parasites . Large genomic constructs integrate at the endogenous locus by homologous recombination, but cDNA-derived sequences lacking long stretches of contiguous genomic DNA (due to intron excision) typically integrate into chromosomal DNA by nonhomologous recombination . Nonhomologous integration occurs effectively at random; and coupled with the high frequency of transformation, this allows a large fraction of the parasite genome to be tagged in a single electroporation cuvette . Genomic tagging permits insertional mutagenesis studies conceptually analogous to transposon mutagenesis in bacteria, yeast, Drosophila, etc . In theory (and, thus far, in practice), this allows identification of any gene whose inactivation is not lethal to the haploid tachyzoite form of T . gondii and for which a suitable selection or screen is available . Transformation vectors can be engineered to facilitate rescue of the tagged locus and to include a variety of reporters or selectable markers . Genetic strategies are also possible, using reporters whose function can be assayed by metabolic, visual, or immunological screens to "trap" genes that are activated (or inactivated) under various conditions of interest . Chem Biol, 1995 Aug, 2(8), 553 - 61 Determination of the structure of exochelin MN, the extracellular siderophore from Mycobacterium neoaurum; Sharman GJ et al.; BACKGROUND: Siderophores are compounds produced by bacteria to acquire iron . Exochelin MN, the extracellular siderophore from Mycobacterium neoaurum, is of particular interest because it has been shown to transport iron into M . leprae, which is responsible for the disease leprosy . Exochelins from other species cannot mediate iron transport in M . leprae, suggesting a specific uptake mechanism involving exochelin MN . We set out to determine the structure of exochelin MN and identify the features of the molecule that may account for this specificity . RESULTS: The structure of exochelin MN was elucidated by a combination of techniques including nuclear magnetic resonance, mass spectrometry, derivatization and gas chromatography . Exochelin MN is a peptide, containing the unusual amino acid beta-hydroxyhistidine and an unusual N-methyl group . The peptide coordinates iron(III) octahedrally using its two cis-hydroxamate groups plus the hydroxyl and imidazole nitrogen of the beta-hydroxyhistidine . The three-dimensional structure of the hexadentate exochelin/gallium complex was deduced from NMR data . CONCLUSIONS: Exochelin MN has some structural features in common with other siderophores, but has a unique three-dimensional structure, which is presumably important for its specific activity in M . leprae . Exochelin MN may be a target for drug design in the fight against infection with this pathogen. Fold Des, 1997, 2(5), R71 - 9 Transmuting alpha helices and beta sheets; Dalal S et al.; Protein architecture involves two main secondary structural classes: alpha helices and beta sheets . Some natural proteins alter their fold in response to changes in solution conditions or as a consequence of mutation . Here, we discuss recent attempts to induce such conformational changes by design: specifically, the motivation and success of efforts to change one protein fold into a different one in response to the 'Paracelsus Challenge' . The results of such efforts may provide a better understanding of the