J Biol Chem, 2002 Jun 21, 277(25), 22304 - 13 Epub 2002 Apr 08. Direct involvement of CREB-binding protein/p300 in sequence-specific DNA binding of virus-activated interferon regulatory factor-3 holocomplex; Suhara W et al.; Infections of bacteria and viruses induce host defense reactions known as innate responses including the activation of interferon regulatory factor-3 (IRF-3), critical for the activation of type I interferon system . Upon immediate early signals triggered by the infection, IRF-3 is phosphorylated and a homodimer results . The homodimer complexes with the coactivator CREB-binding protein (CBP)/p300 in the nucleus; thus, holocomplex of IRF-3 competent in DNA binding is generated . We showed CBP/p300 to be indispensable for the DNA binding activity of the holocomplex and to aid the binding through direct interaction with the DNA . We demonstrated that p300 binds with the IRF-3 homodimer via a Q-rich domain and that an intact histone acetyltransferase (HAT) domain is indispensable for the DNA binding of the holocomplex along with a CH3 domain, which connects the HAT and Q-rich domains . These results highlight a novel function of CBP/p300: direct involvement in sequence-specific DNA binding . Furthermore, the critical function of these domains in virus-induced gene activation was demonstrated in vivo by using p300 mutants. Hunan Yi Ke Da Xue Xue Bao, 1999, 24(1), 80 - 2 {Study on etiologic agents of vaginitis}; Shu M et al.; In order to provide scientific basis for preventing and curing vaginitis, etiologic agents were examined in 354 patients with vaginitis and 115 healthy women using rapid and reliable laboratory methods . The inducements of vaginitis were analysed . The results showed that candida was only detected for the normal control group . Bacteria, candida and trichomonas were the commonest causes of vaginitis . With the increasing ages of patients with vaginitis, the incidence rates of all kinds of vaginitis were lowered . Vaginitis is associated with pregnancy, using antibiotics and contraceptive, or sexual activity. Hunan Yi Ke Da Xue Xue Bao, 1999, 24(1), 56 - 8 {The role of the leukocytes in pathogenesis of secondary brain injury}; Hu S et al.; The levels of leukocyte count, neutrophil number, leukocyte adhesiveness, leukocyte releasing oxygen-free radical (O2.-) after phagocytosing bacteria and serum lipid peroxide MDA were determined in the peripheral blood of 40 patients with acute head injury within 48 hours and 22 normal adults . These levels of patients were significantly higher than those of the controls (P < 0.05), and increased with the severity of craniocerebral injury elevation . The leukocyte count and neutrophil number of those patients who died were significantly higher than these of those survived patients (P < 0.05) . It suggested that leukocytes aggravated the secondary brain injury after craniocerebral injury by increasing of leukocyte adhesiveness, and generating and releasing oxygen-free radical, and measurement of leukocyte count and neutrophil number in the peripheral blood could evaluate the prognosis of the patient with craniocerebral injury. Exp Hematol, 2002 Apr, 30(4), 285 - 96 NFkappaB-dependent signaling pathways; Li X et al.; The transcription factor NFkappaB is activated by numerous stimuli . Once NFkappaB is fully activated, it participates in the regulation of various target genes in different cells to exert its biological functions . NFkappaB has often been referred to as a central mediator of the immune response, since a large variety of bacteria and viruses can lead to the activation of NFkappaB, which in turn controls the expression of many inflammatory cytokines, chemokines, immune receptors, and cell surface adhesion molecules . Recent studies have shown that NFkappaB may function more generally as a central regulator of stress responses, since different stressful conditions, including physical stress, oxidative stress, and exposure to certain chemicals, also lead to NFkappaB activation . Furthermore, NFkappaB blocks cell apoptosis in several cell types . Taken together, these findings make it clear that NFkappaB plays an important role in cell proliferation and differentiation . It is the intention of this review to cover the various NFkappaB-dependent signaling pathways, thereby to achieve a better understanding of the mechanisms of NFkappaB activation and the physiological functions of activated NFkappaB. Parasitol Res, 2002 Feb, 88(2), 113 - 7 Recognition of plasmodium falciparum proteins by mannan-binding lectin, a component of the human innate immune system; Klabunde J et al.; The mannan-binding lectin (MBL) is a serum protein, which is involved in the immune defence against viruses, bacteria and parasites . Children who have mutations in the MBL gene that lead to a MBL deficiency are more susceptible to infectious diseases and are more likely to suffer from severe malaria . In this report we investigate the interaction between MBL and the proteins of red blood cells infected with the parasite Plasmodium falciparum . Protein extracts were separated on MBL-sepharose columns . After the elution of bound material, the proteins were detected either by Western blot with human antibodies, or radioactive labelling with 35S-methionine or 3H-glucosamine . MBL recognises proteins of P . falciparum-infected erythrocytes that are immunogenic in humans, parasite-derived and glycosylated . Whether the proteins identified in the different assays are identical remains to be explored . MBL added to in vitro cultures of P . falciparum, however, does not inhibit parasite growth . The positive effect of MBL in the blood of malaria patients could be caused by detoxification of parasite products. J Biol Chem, 2002 Jun 21, 277(25), 22750 - 8 Epub 2002 Apr 04. Biochemical properties of the PsbS subunit of photosystem II either purified from chloroplast or recombinant; Dominici P et al.; The biochemical properties of PsbS protein, a nuclear-encoded Photosystem II subunit involved in the high energy quenching of chlorophyll fluorescence, have been studied using preparations purified from chloroplasts or obtained by overexpression in bacteria . Despite the homology with chlorophyll a/b/xanthophyll-binding proteins of the Lhc family, native PsbS protein does not show any detectable ability to bind chlorophylls or carotenoids in conditions in which Lhc proteins maintain full pigment binding . The recombinant protein, when refolded in vitro in the presence of purified pigments, neither binds chlorophylls nor xanthophylls, differently from the homologous proteins LHCII, CP26, and CP29 that refold into stable pigment-binding complexes . Thus, it is concluded that if PsbS is a pigment-binding protein in vivo, the binding mechanism must be different from that present in other Lhc proteins . Primary sequence analysis provides evidence for homology of PsbS helices I and III with the central 2-fold symmetric core of chlorophyll a/b-binding proteins . Moreover, a structural homology owed to the presence of acidic residues in each of the two lumen-exposed loops is found with the dicyclohexylcarbodiimide/Ca(2+)-binding domain of CP29 . Consistently, both native and recombinant PsbS proteins showed {(14)C}dicyclohexylcarbodiimide binding, thus supporting a functional basis for its homology with CP29 on the lumen-exposed loops . This domain is suggested to be involved in sensing low luminal pH. Curr Opin Microbiol, 2002 Apr, 5(2), 211 - 5 Connections between transcriptional regulation and type III secretion? Miller VL. Type III secretion is a mechanism of protein export in bacteria, by which proteins (termed effectors) are exported from the bacteria to the external environment . One topic that has escaped much attention is the potential link between the secretion of effectors and transcriptional regulation of genes encoding the effectors . The goal of this commentary is to highlight some of the work that has been done and to stimulate additional research. Curr Opin Microbiol, 2002 Apr, 5(2), 154 - 9 The biological roles of trans-translation; Withey JH et al.; The unique transfer-messenger RNA (tmRNA) molecule has been identified in all bacterial species examined, suggesting that its action confers an important survival advantage to bacteria . Acting both as a tRNA and an mRNA, in a process known as trans-translation, tmRNA adds a short peptide tag to undesirable proteins . Trans-translation plays at least two physiological roles: removing ribosomes stalled upon mRNA, and targeting the resulting truncated proteins for degradation by proteases . The first of these roles is required for all known activities of tmRNA, whereas the second may be dispensed with in most cases with little biological effect . However, tmRNA-targeted proteolysis may be important for fine-tuning expression of certain genes by altering the concentration of regulatory proteins . Here, we review recent literature that addresses the biological functions of tmRNA. Bioorg Med Chem Lett, 2002 Apr 22, 12(8), 1229 - 31 Self-assembly of synthetic zinc chlorins in a silicate micelle prepared by sol-gel process; Saga Y et al.; Zinc methyl 3-devinyl-3-hydroxymethyl-pyropheophorbide-a (1), a good model compound of light-harvesting pigments of green photosynthetic bacteria, formed self-aggregates in the presence of octadecyltriethoxysilane and tetraethoxysilane in an aqueous solution to exhibit visible absorption spectra similar to the natural antenna . Base-catalyzed cross-linked polymerization of the additive Si-ORs (R=ethyl and H) afforded the formation of a siloxane network (Si-O-Si) on the surface of the self-assemblies of 1 . The resulting microcapsules were stable to tolerate the deaggregation to monomeric 1 by addition of surfactant Triton X-100 more largely than the corresponding micelles before polymerization. Biotechnol Prog, 2002 Mar-Apr, 18(2), 193 - 200 Modeling of growth, lactate consumption, and volatile fatty acid production by Megasphaera elsdenii cultivated in minimal and complex media; Soto-Cruz O et al.; Use of the Pirt and Luedeking-Piret equations permits the determination of the effect of medium composition on the metabolic patterns of Megasphaera elsdenii grown in minimal and complex media with lactate as the major carbon source . To establish the significance of the parameters involved in the Pirt and Luedeking-Piret equations, a quantitative statistical criterion was proposed . In the complex medium, lactate was completely used for growth and product formation, whereas in the minimal medium a fraction of the energy obtained from lactate was used for maintenance purposes . Modeling of VFA production by the Luedeking-Piret equation showed that, independent of the type of medium, acetate and propionate are growth-associated products, while butyrate and valerate are only partially growth-associated . The growth-associated products are related to energy-yielding metabolism and the non-growth-associated products are related to the consumption of reducing equivalents. Hum Mutat, 2002 Apr, 19(4), 343 - 60 Analysis of SNPs and other genomic variations using gel-based chips; Kolchinsky A et al.; Application of microarrays for the analysis of point mutations and SNPs in genomic DNAs is currently under intensive development . Various technologies are being investigated, employing enzymatic, chemical, and physical tools {for review, see Tillib and Mirzabekov, 2001} . Our current approach is based on the use of IMAGE chips (immobilized microarrays of gel elements) consisting of an array of gel pads attached to a hydrophobic glass surface . The gel pads range in size from picoliters to nanoliters and are used for immobilization of oligonucleotide probes, as well as miniature test tubes for chemical or enzymatic reactions with tethered compounds . Nucleic acids are hybridized, fractionated, modified, and subjected to enzymatic reactions inside the pads . All steps of sequence analysis (PCR-amplification, activation or release of primers and products, DNA extension, hybridization, and reading of the results) can be performed within the same pad . A flexible and inexpensive technology platform enables one to monitor processes in the arrays in both real time and steady-state . Identification of SNPs, microsequencing, and other specific tasks are easily performed . In particular, stacking interactions with short oligonucleotides enhance the capability of high-throughput screening . The IMAGE chips can be analyzed using a variety of equipment, from a dedicated multi-color fluorescent microscope or MALDI-spectrometer to an inexpensive portable analyzer suitable for field conditions . Customized gel-based chips were successfully used for screening of SNPs in a broad range of biologically meaningful genes . Mol Reprod Dev, 2002 May, 62(1), 37 - 46 Molecular cloning and characterization of fox testis kinectin; Xu J et al.; Kinectin was isolated and characterized from a fox testis cDNA library using a monoclonal antibody (FTA-1) raised against testis surface proteins . The cDNA sequence of 4,479 nucleotides encodes an ORF of 1,330 amino acids (aa) with high homology to mouse, human, and chicken kinectins (GenBank Accession Number AF095786) . Southern analysis was used to show that genes homologous to kinectin are present in several mammal species and in at least one marsupial, but not in bacteria . Alternatively spliced forms of fox kinectin were identified, and one of these is uniquely expressed in brain and spleen tissues . Kinectin expression was highest in testis relative to other tissues examined . Sequence analysis and comparisons between species revealed that kinectin encodes multiple alpha-helical coiled coils predicted to form dimers, and is, therefore, likely to exist as a dimer . The results presented in this article suggest that kinectin is required for spermatogenesis, but is not a likely candidate for use in immunocontraceptive vaccines . Biologist (London), 2002 Apr, 49(2), 58 - 62 Proteomics: the protein revolution; Cash P; Proteomics, the latest scientific buzzword and research field, represents a milestone in biological research, as proteins return to centre stage . But what has led to this protein renaissance and why has proteomics become a key research tool in the post-genomic era? The answers to these questions lie in the huge progress that has been made in the area of genomics. Microbiology, 2002 Apr, 148(Pt 4), 961 - 71 Physiological consequences associated with overproduction of Mycobacterium tuberculosis FtsZ in mycobacterial hosts; Dziadek J et al.; The ftsZ gene of Mycobacterium tuberculosis H37Rv has been characterized as the first step in determining the molecular events involved in the cell division process in mycobacteria . Western analysis revealed that intracellular levels of FtsZ are growth phase dependent in both M . tuberculosis and Mycobacterium smegmatis . Unregulated expression of M . tuberculosis ftsZ from constitutive hsp60 and dnaA promoters in M . tuberculosis hosts resulted in lethality whereas expression from only the hsp60 promoter was toxic in M . smegmatis hosts . Expression of ftsZ from the dnaA promoter in M . smegmatis resulted in approximately sixfold overproduction and the merodiploids exhibited slow growth, an increased tendency to clump and filament, and in some cases produced buds and branches . Many of the cells also contained abnormal and multiple septa . Expression of ftsZ from the chemically inducible acetamidase promoter in M . smegmatis hosts resulted in approximately 22-fold overproduction of FtsZ and produced filamentous cells, many of which lacked any visible septa . Visualization of the M . tuberculosis FtsZ tagged with green fluorescent protein in M . smegmatis by fluorescence microscopy revealed multiple fluorescent FtsZ foci, suggesting that steps subsequent to the formation of organized FtsZ structures but prior to septum formation are blocked in FtsZ-overproducing cells . Together these results suggest that the intracellular concentration of FtsZ protein is critical for productive septum formation in mycobacteria. Acta Pharmacol Sin, 2002 Apr, 23(4), 300 - 4 Effect of phorbol esters on activity of Ha-ras gene promoter in HeLa cells; Zhang W et al.; AIM: To study the role of phorbol esters (PMA) in the activity of promoter of Ha-ras in HeLa cells.METHODS: After treatment with PMA, the growth rate of HeLa cells was measured by MTT assay and the expression of Ha-ras gene was detected by reversed transcriptase polymerase chain reaction . The regulation fragment of Ha-ras was constructed into the plasmid enhanced yellow fluorescent protein (EYFP) which did not contain promoter element . The recombinant plasmid pRasEYFP was transient transfected to HeLa cells . Using the EYFP gene as reporter, the activity of promoter of Ha-ras after treatment of PMA was assayed . RESULTS: The growth rate of PMA-treated HeLa cells was markedly reduced compared with untreated cells . The expression of Ha-ras gene was obviously decreased and the activity of promoter of Ha-ras was decreased by 34.0 %and 26.7 % respectively in HeLa cells after treated with PMA (100 microg/L) for 48 h and 72 h . In the meanwhile the activity of protein kinase C (PKC) was decreased by 76.3 % and 73.2 % compared with the control cells, respectively . CONCLUSION: PMA play an important role in regulation of the activity of promoter of Ha-ras in HeLa cells . The molecular mechanism may be through the PKC pathway. Biochem J, 2002 Apr 15, 363(Pt 2), 305 - 11 Modulation of Mycobacterium tuberculosis DnaA protein-adenine-nucleotide interactions by acidic phospholipids; Yamamoto K et al.; The biochemical aspects of the initiation of DNA replication in Mycobacterium tuberculosis are unknown . To understand this process, we overproduced, purified and characterized the recombinant M . tuberculosis DnaA protein . The M . tuberculosis DnaA protein binds the origin of replication (oriC), ATP and ADP, and exhibited weak ATPase activity . ADP, after hydrolysis of ATP, remained strongly associated with DnaA and the exchange of ATP for bound ADP was weak . Vesicles prepared from acidic phospholipids, such as phosphatidylinositol, cardiolipin and phosphatidylglycerol, promoted dissociation of both ADP and ATP, whereas the neutral phospholipid phosphatidylethanolamine did not . The phospholipid-mediated dissociation of ATP was decreased in the presence of the M . tuberculosis oriC, whereas dissociation of ADP was stimulated in the presence of oriC . Acidic phospholipids in micelles, however, were not efficient in dissociating bound nucleotides from DnaA . Together, these results suggest that both polar head groups and membrane bilayer structure play an important role in M . tuberculosis DnaA-adenine-nucleotide interactions . We suggest that initiation of M . tuberculosis oriC involves intimate interactions between DnaA, adenine nucleotides and membrane phospholipids, and the latter helps to ensure that only the ATP form of the DnaA protein interacts continuously with oriC. J Mol Microbiol Biotechnol, 2002 May, 4(3), 249 - 53 Oxygen-regulated expression of genes for pigment binding proteins in Rhodobacter capsulatus; Gregor J et al.; Oxygen is the major external factor affecting the expression of photosynthesis genes in facultatively photosynthetic bacteria . Many investigations over the last years mainly carried out on the closely related species Rhodobacter capsulatus and Rhodobacter sphaeroides have identified a number of proteins involved in the oxygen-regulated signal pathway, in which the RegB/RegA two component system plays a central role . While the RegB/RegA system activates photosynthesis genes under low oxygen tension other proteins like CrtJ and PPBP have a repressing function under high oxygen tension . Additional DNA binding proteins like the integration host factor can modulate the expression of photosynthesis genes . The role of alternative sigma factors in this signal pathway is still unclear. J Med Entomol, 2002 Jan, 39(1), 162 - 72 Density-dependent development of Anopheles gambiae (Diptera: Culicidae) larvae in artificial habitats; Gimnig JE et al.; The growth and development of Anopheles gambiae Giles larvae were studied in artificial habitats in western Kenya . Larvae responded to increasing densities by extending their development time and by emerging as smaller adults, although survival was not significantly affected . Addition of nutrients in the form of cow dung collected near the study site had no impact on larval growth and development . Regression analysis showed that female development time increased by 0.020 d and female dry mass decreased by 0.74 microg with each additional larva . By fitting the data to the pupation window model, the estimated minimum dry mass to achieve pupation was 0.130 mg and the estimated minimum time to pupation was 5 d . The most likely food source for An . gambiae larvae was algal growth, which was significantly reduced by the presence of larvae . Bacterial densities were not significantly affected by the presence of larvae although total bacteria counts were lower at the higher densities indicating they may provide a secondary food source when algal resources are depleted . Similarly, the levels of nitrogen and phosphorus in the habitats were not significantly affected by the presence of larvae although there was evidence of decreasing nitrogen levels occurring with increasing larval densities suggesting that nitrogen may be a limiting resource in the larval environment . The data indicate that competition within the larval environment may indirectly regulate An . gambiae populations by reducing adult body size, which may in turn reduce adult survivorship and fecundity . The potential impact of density-dependent interactions among An . gambiae larvae on the transmission of Plasmodium falciparum is discussed. J Biol Chem, 2002 Jun 21, 277(25), 22677 - 84 Epub 2002 Apr 19. A novel group of oleosins is present inside the pollen of Arabidopsis; Kim HU et al.; In plants, subcellular triacylglycerol granules in seeds (oil bodies) and floral tapetum (tapetosomes) are stabilized by amphipathic structural protein called oleosin . We hereby report a novel group of oleosins that is present inside the pollen of Arabidopsis thaliana . We have used the conserved sequence of oleosins to locate, via the DNA database, all 16 oleosin genes in the Arabidopsis genome . The oleosin genes can be divided into three groups according to their sequences and tissue-specific expressions, as probed by RNA blot hybridization and reverse transcriptase-PCR . The first group includes eight genes specifically expressed in the floret tapetum . The second group includes five genes specifically expressed in maturing seeds . The third, novel group includes three genes expressed in both maturing seeds and floral microspores, which will become pollen . Transgenic study using the promoter of one of these genes attached to a reporter gene has provided corroborative evidence for the specific expression of the gene in the microspores in the florets . One of the pollen oleosins can be identified by microsequencing and specific immunoblotting . Pollen oleosins synthesized by recombinant bacteria can collaborate with phospholipids in stabilizing reconstituted oil bodies . Thus, pollen has oleosins to stabilize the abundant subcellular oil bodies . Seed oil bodies and floret tapetosomes have been isolated from the miniature Arabidopsis plants, and the success indicates that the organelles can be subjected to future biochemical and genetic studies. Traffic, 2002 Apr, 3(4), 279 - 88 Selective delivery of secretory cargo in Golgi-derived carriers of nonepithelial cells; Rustom A et al.; In epithelial cells, soluble cargo proteins destined for basolateral or apical secretion are packaged into distinct trans-Golgi network-derived transport carriers . Similar carriers, termed basolateral- and apical-like, have been observed in nonepithelial cells using ectopically expressed membrane marker proteins . Whether these cells are capable of selectively packaging secretory proteins into distinct carriers is still an open question . Here, we have addressed this issue by analyzing the packaging and transport of secretory human chromogranin B fusion proteins using a green fluorescent protein-based high-resolution, dual-color imaging technique . We were able to show that these secretory markers were selectively packaged at the Golgi into tubular/vesicular-like transport carriers containing basolateral membrane markers, resulting in extensive cotransport . In contrast, deletion mutants of the human chromogranin B fusion proteins lacking an N-terminal loop structure were efficiently transported in both basolateral- and apical-like carriers, the latter displaying a spherical morphology . Similarly, in polarized epithelial cells, the human chromogranin B fusion protein was secreted basolaterally and the loop-deleted analogue into both the basolateral and apical medium . These findings suggest that nonepithelial cells, like their epithelial counterparts, possess a sorting machinery capable of selective packaging of secretory cargo into distinct types of carriers. Mol Microbiol, 2002 Feb, 43(4), 971 - 80 c-Src/Lyn kinases activate Helicobacter pylori CagA through tyrosine phosphorylation of the EPIYA motifs; Stein M et al.; The human pathogen Helicobacter pylori colonizes the mucous layer of the stomach . During parasitic infection, freely swimming bacteria adhere to the gastric epithelial cells and trigger intracellular signalling pathways . This process requires the translocation of the effector protein CagA into the host cell through a specialized type IV secretion system encoded in the cag pathogenicity island . Following transfer, CagA is phosphorylated on tyrosine residues by a host cell kinase . Here, we describe how the tyrosine phosphorylation of CagA is restricted to a previously identified repeated sequence called D1 . This sequence is located in the C-terminal half of the protein and contains the five-amino-acid motif EPIYA, which is amplified by duplications in a large fraction of clinical isolates . Tyrosine phosphorylation of CagA is essential for the activation process that leads to dramatic changes in the morphology of cells growing in culture . In addition, we observed that two members of the src kinases family, c-Src and Lyn, account for most of the CagA-specific kinase activity in host cell lysates . Thus, CagA translocation followed by tyrosine phosphorylation at the EPIYA motifs promotes a growth factor-like response with intense cytoskeletal rearrangements, cell elongation effects and increased cellular motility. Mol Microbiol, 2002 Feb, 43(4), 945 - 60 Plasticity of a transcriptional regulation network among alpha-proteobacteria is supported by the identification of CtrA targets in Brucella abortus; Bellefontaine AF et al.; CtrA is a master response regulator found in many alpha-proteobacteria . In Caulobacter crescentus and Sinorhizobium meliloti, this regulator is essential for viability and is transcriptionally autoregulated . In C . crescentus, it is required for the regulation of multiple cell cycle events, such as DNA methylation, DNA replication, flagella and pili biogenesis and septation . Here, we report the characterization of the ctrA gene homologue in the alpha2-proteobacteria Brucella abortus, a facultative intracellular pathogen responsible for brucellosis . We detected CtrA expression in the main Brucella species, and its overproduction led to a phenotype typical of cell division defect, consistent with its expected role . A purified B . abortus CtrA recombinant protein (His6-CtrA) was shown to protect the B . abortus ctrA promoter from DNase I digestion, suggesting transcriptional autoregulation, and this protection was enhanced under CtrA phosphorylation on a conserved Asp residue . Despite the similarities shared by B . abortus and C . crescentus ctrA, the pathway downstream from CtrA may be distinct, at least partially, in both bacteria . Indeed, beside ctrA itself, only one (the ccrM gene) out of four B . abortus homologues of known C . crescentus CtrA targets is bound in vitro by phosphorylated B . abortus CtrA . Moreover, further footprinting experiments support the hypothesis that, in B . abortus, CtrA might directly regulate the expression of the rpoD, pleC, minC and ftsE homologues . Taken together, these results suggest that, in B . abortus and C . crescentus, similar cellular processes are regulated by CtrA through the control of distinct target genes . The plasticity of the regulation network involving CtrA in these two bacteria may be related to their distinct lifestyles. Ned Tijdschr Tandheelkd, 1998 Nov, 105(11), 416 - 8 {Female hormones and oral health}; Meijer van Putten JB; A common oral manifestation of elevated levels of the ovarian hormones estrogen and progesterone, as seen in pregnancy or oral contraceptive usage, is an increase in gingival inflammation . Estrogen and progesterone probably induce a physiologic vascular phenomenon with swelling and redness . Furthermore, these hormones alter the microenvironment of the oral bacteria so as to promote their growth and cause shifts in their populations . The resulting gingivitis can be avoided or at least minimized by establishing low plaque levels at the beginning of pregnancy or at the start of oral contraceptive therapy. J Biol Chem, 2002 Jun 14, 277(24), 22085 - 92 Epub 2002 Apr 01. Autoubiquitination of the BRCA1*BARD1 RING ubiquitin ligase; Chen A et al.; The RING finger of BRCA1 confers ubiquitin ligase activity that is markedly enhanced when complexed with another RING-containing protein, BARD1, and is required for the function of this tumor suppressor protein in protecting genomic integrity . Here, we report that co-expression of BRCA1-(1-639) and BARD1 in bacteria can assemble a potent ubiquitin ligase activity . Purified BRCA1-(1-639)*BARD1 stimulated the Ubc5c-mediated monoubiquitination of histone H2A/H2AX in vitro, suggesting a possible role for BRCA1*BARD1 in modifying chromatin structure . Moreover, the truncated BRCA1*BARD1 complex exhibited efficient autoubiquitination activity in vitro capable of assembling non-lysine 48-linked polyubiquitin chains on both BRCA1-(1-639) and BARD1 . When co-expressed in cells by transient transfection, the recombinant BRCA1-(1-300).BARD1 complex was found to be associated with polyubiquitin chains, suggesting that BRCA1-(1-300)*BARD1 was ubiquitinated in vivo as well . These results raise the possibility that BRCA1*BARD1 acts to assemble non-lysine 48-linked polyubiquitin chains that may serve as part of a signaling platform required for coordinating DNA repair-related events. Transpl Infect Dis, 2001, 3 Suppl 2, 49 - 56 Use of cytomegalovirus immune globulin and ganciclovir for the prevention of cytomegalovirus disease in lung transplantation; Zamora MR; Cytomegalovirus (CMV) infection and disease continue to be significant causes of morbidity and mortality in lung transplant recipients . The potential benefits of CMV prophylaxis extend beyond prevention of the immediate CMV infection to potentially preventing CMV-associated complications, including superinfection due to Aspergillus bacteria, and other opportunistic infections, and bronchiolitis obliterans syndrome (BOS) . Longer courses of prophylactic intravenous (IV) ganciclovir, sequential IV/oral therapy, addition of intravenous CMV immune globulin (CMV-IGIV), surveillance tests, and investigation of the role of hypogammaglobulinemia are a few of the strategies and issues being evaluated to improve CMV prophylaxis and, consequently, graft and patient survival. Z Naturforsch {C}, 2002 Jan-Feb, 57(1-2), 161 - 71 Chemical defense and antifouling activity of three Mediterranean sponges of the genus Ircinia; Tsoukatou M et al.; The defense roles and the antifouling activity of the organic extracts and the major metabolites of the sponges Ircinia oros, I . variabilis and I . spinosula were investigated . The antifeedant activity was tested in experimental aquaria on the generalist predator fish Thalassoma pavo as well as in coastal ecosystems rich in fishes . Some of the major metabolites exhibited high levels of antifeedant activity . The antifouling activity was tested in laboratory assays, against representatives of the major groups of fouling organisms (marine bacteria, marine fungi, diatoms, macroalgae and mussels) . All extracts showed promising levels of activity . As was expected, no single extract was active in all tests and some fractions that were effective against one organism showed little or no activity against the others . The high but variable level of antifouling activity in combination with the absence of toxicity (tested on the development of oyster and sea urchin larvae) shows the potential of these metabolites to become ingredients in environmentally friendly antifouling preparations. World J Gastroenterol, 2002 Apr, 8(2), 323 - 7 Infrared thermoimages display of body surface temperature reaction in experimental cholecystitis; Zhang D et al.; AIM: To display the thermoimages of the body surface in experimental cholecystitis, to observe the body surface temperature reaction in visceral disorders, and to study if the theory of body surface-viscera correlation is true and the mechanism of temperature changes along the meridians . METHODS: By injecting bacteria suspension into the stricture bile duct and gallbladder, 21 rabbits were prepared as acute pyogenic cholangiocholecystitis models, with another 8 rabbits prepared by the same process except without injection of bacteria suspension as control . The body surface infrared thermoimages were continuously observed on the hair shaven rabbit skin with AGA-782 thermovision 24h before, 1-11 d after and (2,3 wk) 4 wk after the operation with a total of over 10 records of thermoimages . RESULTS: Twelve cases out of 21 rabbits with cholecystitis revealed bi-lateral longitudinal high temperature lines in its trunk; with negative findings in the control group . The high-temperature line appeared on d1-d2, first in the right trunk, after the preparation of the model, about 7 d after the model preparation, the lines appeared at the left side too, persisting for 4 wk . The hyper-temperature line revealed 1.1-2.7 degrees higher than before the model preparation, 0.7-2.5 degrees higher than the surrounding skin . The length of the high temperature line might reach a half length of the body trunk, or as long as the whole body itself . CONCLUSION: The appearance of the longitudinal high temperature lines at the lateral aspects of the trunk in the experimental group is directly bound up with the experimental animals pyogenic cholecystitis, with its running course quite similar to that of the Gallbladder Channel of Foot Shaoyang, but different to the zones of hyperalgesia and site of referred pain in cholecystitis. Planta, 2002 Feb, 214(4), 497 - 504 Jasmonate-related mutants of Arabidopsis as tools for studying stress signaling; Berger S; Jasmonates are naturally occurring signal compounds that regulate plant growth and development, and are involved in plant responses to several environmental stress factors . The mode of action of jasmonates has been investigated traditionally by analysis of the effects of exogenous application of these compounds, including identification of jasmonate-responsive genes and determination of their expression and responsive promoter elements . In addition, jasmonate biosynthesis has been studied by identification of biosynthetic enzymes, use of inhibitors and determination of endogenous jasmonate levels . Recently, several mutants defective in jasmonate biosynthesis and signaling have been isolated and their phenotypes shed new light on the role of jasmonates and jasmonate signaling in plant responses to pathogens, insects and ozone. Comp Med, 2001 Dec, 51(6), 538 - 44 Mycobacterium avium subspecies paratuberculosis triggers intestinal pathophysiologic changes in beige/scid mice; Mutwiri GK et al.; We investigated whether infection of beige/scid mice with Mycobacterium avium subspecies paratuberculosis can induce intestinal pathophysiologic changes . Six-week-old beige/scid mice were inoculated intraperitoneally with M . paratuberculosis, then were killed 32 weeks after inoculation when the small intestine was evaluated for physiologic and morphologic abnormalities . All infected mice developed clinical disease . The lamina propria of the intestine from infected mice was mildly infiltrated with mononuclear cells containing acid-fast bacteria, and had significantly increased villus width . In vitro physiologic studies in Ussing chambers indicated that M . paratuberculosis infection caused significant abnormalities in intestinal transport parameters . Baseline short circuit current and potential difference were abnormally high in tissues from infected, compared with control mice, indicative of increased ion secretion . Baseline conductance was significantly decreased in infected mice, suggesting that intestinal tissue from infected mice was less permeable to ions . The change in short circuit current following transmural electrical and glucose stimulation was significantly reduced in intestines from infected mice, suggesting that inflamed intestine had neural and/or epithelial cell damage . We conclude that infection of beige/scid mice with M . paratuberculosis triggers significant intestinal pathophysiologic changes consistent with chronic inflammation . These functional abnormalities may contribute to the pathogenesis of the wasting syndrome seen in bovids with paratuberculosis . This animal model provides evidence that T cell-independent mechanisms are sufficient to cause mucosal pathophysiologic changes and inflammation in response to a specific pathogen, and may be of relevance to inflammatory bowel disease in humans. Comp Med, 2001 Aug, 51(4), 357 - 60 Coxiella burnetii infection in C.B-17 scid-bg mice xenotransplanted with fetal bovine tissue; Criley JM et al.; Two from a group of approximately 50 C.B-17 scid-bg mice were examined because of lethargy, dehydration, and rough coat . Three months prior to development of clinical signs of disease, mice of this study had been surgically implanted with fetal bovine liver, thymus, and lymph node . At necropsy, marked splenomegaly and mild hepatomegaly were observed in both animals . Large areas of necrosis and inflammation, with associated intracytoplasmic granular basophilic inclusions, were observed in histologic sections of multiple organs . Aerobic and anaerobic culturing of the liver yielded negative results . Six months after the initial case, four more reconstituted scid-bg mice from a different fetal donor had identical clinical, gross, and histologic signs of disease . To determine whether the basophilic inclusions represented an infective agent, 4-month-old immune-naive C.B-17 scid-bg mice were inoculated intraperitoneally with a liver and spleen homogenate from an affected mouse . Two weeks after inoculation, mice developed clinical signs of disease and lesions identical to those seen in the signal mice . On ultrastructural examination of the liver, pleomorphic bacteria were found in large cytoplasmic vacuoles of hepatocytes . Bacterial DNA was amplified from the liver, using primers that amplify a segment of the 16S rRNA gene from many bacterial species . Sequencing of the polymerase chain reaction (PCR) product revealed gene sequence identical to that of Coxiella burnetii, the agent of Q-fever . These results highlight the need to consider infective agents of the donor species when working with xenografted animals. Phytomedicine, 2002 Jan, 9(1), 33 - 40 Comparison of antioxidative capacities and inhibitory effects on cholesterol biosynthesis of quercetin and potential metabolites; Glasser G et al.; The flavonol quercetin is known to be rapidly metabolized after ingestion by enterocytes and bacteria in the intestinal tract which may influence the biological, e.g . antioxidative potency of this compound . Therefore, quercetin and several of its possible metabolites were compared with regard to their antioxidant activity and their capacity to inhibit hepatocellular cholesterol biosynthesis . Using the 2,2,-diphenylpicrylhydrazyl radical scavenger assay, all compounds with an ortho diphenolic structure acted as strong antioxidants . In contrast, in a cellular assay focusing on lipid peroxidation in cultured rat hepatocytes challenged with tert.-butylhydroperoxide only the lipophilic compounds quercetin and 3,4-dihydroxytoluene were active . Concerning the inhibition of cholesterol biosynthesis, 3,4-dihydroxytoluene surprisingly mimicked the effect of quercetin in primary rat hepatocytes, but much less so in HepG2 cells . All other metabolites were almost ineffective in both cell types . These results suggest that some of the biological functions of flavonoids detectable by in vitro assays may persist in vivo as long as comparably potent metabolites are systemically present. BMC Oral Health . 2002;2(1):2. Elevated antibody to D-alanyl lipoteichoic acid indicates caries experience associated with fluoride and gingival health; Levine M et al.; : BACKGROUND: Acidogenic, acid-tolerant bacteria induce dental caries and require D-alanyl glycerol lipoteichoic acid (D-alanyl LTA) on their cell surface . Because fluoride inhibits acid-mediated enamel demineralization, an elevated antibody response to D-alanyl LTA may indicate subjects with more acidogenic bacteria and, therefore, an association of DMFT with fluoride exposure and gingival health not apparent in low responders . METHODS: Cluster analysis was used to identify low antibody content . Within low and high responders (control and test subjects), the number of teeth that were decayed missing and filled (DMFT), or decayed only (DT) were regressed against fluoride exposure in the water supply and from dentrifice use . The latter was determined from gingival health: prevalences of plaque (PL) and bleeding on probing (BOP), and mean pocket depth (PD) . Age was measured as a possible confounding cofactor . RESULTS: In 35 high responders, DMFT associated with length of exposure to fluoridated water (F score), PL and BOP (R2 = 0.51, p < 0.001), whereas in 67 low D-ala-IgG responders, DMFT associated with PL, age, and PD (R2 = 0.26, p < 0.001) . BOP correlated strongly with number of 7 7 decayed teeth (DT) in 54 high responders (R2 = 0.57, p < 0.001), but poorly in 97 low responders (R2 = 0.12, p < 0.001) . The strength of the PD association with DMFT, or of BOP with DT, in high responders significantly differed from that in low responders (p < 0.05) . CONCLUSION: Caries associates with gingival health and fluoridated water exposure in high D-alanyl LTA antibody responders. Science, 2002 Mar 29, 295(5564), 2452 - 6 Visualization of a Ran-GTP gradient in interphase and mitotic Xenopus egg extracts; Kalab P et al.; The small guanosine triphosphatase Ran is loaded with guanosine triphosphate (GTP) by the chromatin-bound guanine nucleotide exchange factor RCC1 and releases import cargoes in the nucleus during interphase . In mitosis, Ran-GTP promotes spindle assembly around chromosomes by locally discharging cargoes that regulate microtubule dynamics and organization . We used fluorescence resonance energy transfer-based biosensors to visualize gradients of Ran-GTP and liberated cargoes around chromosomes in mitotic Xenopus egg extracts . Both gradients were required to assemble and maintain spindle structure . During interphase, Ran-GTP was highly enriched in the nucleoplasm, and a steep concentration difference between nuclear and cytoplasmic Ran-GTP was established, providing evidence for a Ran-GTP gradient surrounding chromosomes throughout the cell cycle. J Biol Chem, 2002 Jun 7, 277(23), 20431 - 7 Epub 2002 Mar 28. TLR4 and MD-2 expression is regulated by immune-mediated signals in human intestinal epithelial cells; Abreu MT et al.; The normal intestinal epithelium is not inflamed despite contact with a high density of commensal bacteria . Intestinal epithelial cells (IEC) express low levels of TLR4 and MD-2 and are lipopolysaccharide (LPS)-unresponsive . We hypothesized that immune-mediated signals regulate the expression of TLR4 and MD-2 in IEC . Expression of TLR4 and MD-2 was examined in normal colonic epithelial cells or intestinal epithelial cell lines . The effect of the cytokines interferon (IFN)-gamma, IFN-alpha, and tumor necrosis factor-alpha (TNF-alpha) on TLR4 and MD-2 expression was examined by reverse transcription-PCR and Western blot . NF-kappaB transcriptional activation and interleukin-8 secretion were used as measures of LPS responsiveness . Native colonic epithelial cells and IEC lines express a low level of TLR4 and MD-2 mRNA . IFN-gamma regulates MD-2 expression in both IEC lines, whereas IFN-gamma and TNF-alpha regulate TLR4 mRNA expression in IEC lines . Pre-incubation with IFN-gamma and/or TNF-alpha sensitizes IEC to LPS-dependent interleukin-8 secretion . To examine MD-2 transcriptional regulation, we cloned a 1-kb sequence proximal to the MD-2 gene translational start site . This promoter directed expression of a reporter gene in endothelial cells and IEC . IFN-gamma positively regulated MD-2 promoter activity in IEC . Co-expression of a STAT inhibitor, SOCS3, blocked IFN-gamma-mediated MD-2 promoter activation . T cell-derived cytokines lead to increased expression of TLR4 and MD-2 and LPS-dependent pro-inflammatory cytokine secretion in IEC . IFN-gamma regulates expression of the critical TLR4 co-receptor MD-2 through the Janus tyrosine kinase-STAT pathway . Th1 cytokines may initiate or perpetuate intestinal inflammation by altering toll-like receptor expression and bacterial reactivity. BMJ . 2002 Mar 30;324(7340):763. Synergism between allergens and viruses and risk of hospital admission with asthma: case-control study; Green RM et al.; OBJECTIVE: To investigate the importance of sensitisation and exposure to allergens and viral infection in precipitating acute asthma in adults resulting in admission to hospital . DESIGN: Case-control study . SETTING: Large district general hospital . PARTICIPANTS: 60 patients aged 17-50 admitted to hospital over a year with acute asthma, matched with two controls: patients with stable asthma recruited from the outpatient department and patients admitted to hospital with non-respiratory conditions (inpatient controls) . Main outcome measures: Atopic status (skin testing and total and specific IgE), presence of common respiratory viruses and atypical bacteria (polymerase chain reaction), dust samples from homes, and exposure to allergens (enzyme linked immunosorbent assay (ELISA): Der p 1, Fel d 1, Can f 1, and Bla g 2) . RESULTS: Viruses were detected in 31 of 177 patients . The difference in the frequency of viruses detected between the groups was significant (admitted with asthma 26%, stable asthma 18%, inpatient controls 9%; P=0.04) . A significantly higher proportion of patients admitted with asthma (66%) were sensitised and exposed to either mite, cat, or dog allergen than patients with stable asthma (37%) and inpatient controls (15%; P<0.001) . Being sensitised and exposed to allergens was an independent associate of the group admitted to hospital (odds ratio 2.3, 95% confidence interval 1.0 to 5.4; P=0.05), whereas the combination of sensitisation, high exposure to one or more allergens, and viral detection considerably increased the risk of being admitted with asthma (8.4, 2.1 to 32.8; P=0.002) . CONCLUSIONS: Allergens and viruses may act together to exacerbate asthma. Clin Chim Acta, 2002 May 7, 319(1), 57 - 62 Assessing nitrate metabolism in the intestinal tract by measuring breath nitric oxide and nitrous oxide, and its clinical significance; Mitsui T et al.; BACKGROUND: The toxicity of dietary nitrate (NO3-) is controversial . One reason is nitrate metabolism in the intestine is so complicated that it is far from fully understood . There is no study measuring breath nitric oxide (NO) and nitrous oxide (N2O) after ingesting vegetables and high-nitrate food at the same time . METHODS: Breath samples from 10 healthy young and 10 healthy old subjects were collected at 15-min intervals for 5 h after ingestion of 100 g of lettuce and during fasting (control) . Breath NO and N2O were analyzed by a chemiluminescence and an IR-PAS analyzer respectively . RESULTS: N2O maximum concentration and excretions increased significantly after ingesting lettuce in each group {303 (30) vs . 750 (81) ppb, 771 (72) vs . 1668 (146) microg in young; 442 (52) vs . 1092 (109) ppb, 1088 (125) vs . 2100 (183) microg in old subjects; mean (SE), P<0.01}, while NO did not . In addition, breath NO was strongly influenced by ambient NO, which varied greatly . N2O maximum level in old subjects after ingesting lettuce was higher than that of young subjects (750 vs . 1092 ppb, P<0.05), and significantly higher N2O concentration levels were seen at 30, 45, 60, and 105 min in old subjects . CONCLUSIONS: A large amount of N2O produced in the intestine and normal nitrate intake do not influence the breath NO concentration, probably due to its relatively small production . Higher maximum N2O concentration after ingesting lettuce in old subject is probably because more bacteria, which rapidly reduce dietary nitrate in the upper intestinal tract, inhabit the gut in old age . Our results suggested that breath N2O is a useful noninvasive maker to estimate dietary nitrate reduction in the intestinal tract. Parassitologia, 2001 Sep, 43(3), 143 - 6 Molecular identification of Borrelia valaisiana and HGE-like Ehrlichia in Ixodes ricinus ticks sampled in north-eastern Italy: first report in Veneto region; Favia G et al.; PCR amplification was applied to screen the presence of both Borrelia burgdorferi s.l . and Ehrlichia species in pools of field-collected Ixodes ricinus ticks . The specimens so far analysed (n = 55), grouped in 11 pools, were sampled in Feltre area (Veneto region, NE Italy) . Five pools proved positive for B . valaisiana (45%) and one of them (9%) was also positive for Ehrlichia, that was further characterised as a HGE-like Ehrlichia . This is the first report of the two bacteria in the Veneto region . The pool positive for both pathogens was used to adjust a multiplex PCR assay, which allowed the detection and identification of both parasites in a single experiment . The advantages offered by this assay, when standardised, will substantially broaden the perspectives of ecological and epidemiological investigations on animal/human Lyme disease and ehrlichiosis, greatly facilitating disease surveillance and control programs. Environ Mol Mutagen, 2002, 39(2-3), 143 - 9 Mutagens in human breast lipid and milk: the search for environmental agents that initiate breast cancer; Phillips DH et al.; Epidemiological studies indicate the involvement of environmental factors in the etiology of breast cancer, but have not provided clear indications of the nature of the agents responsible . Several environmental carcinogens are known to induce mammary tumors in rodents, and the abundance of adipose tissue in the human breast suggests that the epithelial cells, from which breast tumors commonly arise, could be exposed to lipid-soluble carcinogens sequestered by the adipose tissue . In this report we review our studies in which we have examined human mammary lipid, obtained from elective reduction mammoplasties from healthy donors, and human milk from healthy mothers, for the presence of components with genotoxic activity in several in vitro assays . A significant proportion of lipid extracts induced mutations in bacteria and micronuclei in mammalian cells . They also caused DNA damage, detected as single-strand breaks in the alkaline single-cell gel electrophoresis (comet) assay, in both the MCL-5 cell line and in primary cultures of human mammary epithelial cells . Genotoxic activity was also found in a significant proportion of extracts of human breast milk . Viable cells recovered from milk samples showed evidence of DNA damage and were susceptible to comet formation by genotoxic agents in vitro . Genotoxic activity was found to be less prevalent in milk samples from countries of lower breast cancer incidence (the Far East) compared with that in samples from the UK . The agents responsible for the activity in milk appear to be moderately polar lipophilic compounds and of low molecular weight . Identification of these agents and their sources may hold clues to the origins of breast cancer . Mol Biol Evol, 2002 Apr, 19(4), 456 - 61 A new subfamily of major intrinsic proteins in plants; Johanson U et al.; The major intrinsic proteins (MIPs) form a large protein family of ancient origin and are found in bacteria, fungi, animals, and plants . MIPs act as channels in membranes to facilitate passive transport across the membrane . Some MIPs allow small polar molecules like glycerol or urea to pass through the membrane . However, the majority of MIPs are thought to be aquaporins (AQPs), i.e., they are specific for water transport . Plant MIPs can be subdivided into the plasma membrane intrinsic protein, tonoplast intrinsic protein, and NOD26-like intrinsic protein subfamilies . By database mining and phylogenetic analyses, we have identified a new subfamily in plants, the Small basic Intrinsic Proteins (SIPs) . Comparisons of sequences from the new subfamily with conserved amino acid residues in other MIPs reveal characteristic features of SIPs . Possible functional consequences of these features are discussed in relation to the recently solved structures of AQP1 and GlpF . We suggest that substitutions at conserved and structurally important positions imply a different substrate specificity for the new subfamily. J Biol Chem, 2002 Jun 7, 277(23), 20372 - 8 Epub 2002 Mar 27. Structure/function relationships in OxlT, the oxalate/formate antiporter of Oxalobacter formigenes: assignment of transmembrane helix 2 to the translocation pathway; Ye L et al.; We constructed a single cysteine panel encompassing transmembrane helix two (TM2) of OxlT, the oxalate/formate antiporter of Oxalobacter formigenes . Among the 21 positions targeted, cysteine substitution identified one (phenylalanine 59) as essential to OxlT expression and three (glutamine 56, glutamine 66, and serine 69) as potentially critical to OxlT function . By probing membranes with a bulky hydrophilic probe (Oregon Green maleimide) we also located a central inaccessible core of at least eight residues in length, extending from leucine 61 to glycine 68 . Functional assays based on reconstitution of crude detergent extracts showed that of single cysteine mutants within the TM2 core only the Q63C variant was substantially (> or =95%) inhibited by thiol-specific agents (carboxyethyl methanethiosulfonate and ethylsulfonate methanethiosulfonate) . Subsequent analytical work using the purified Q63C protein showed that inhibition by ethylsulfonate methanethiosulfonate was blocked by substrate and that the concentration dependence of such substrate protection occurred with a binding constant of 0.16 mm oxalate, comparable with the Michaelis constant observed for oxalate transport (0.23 mm) . These findings lead us to conclude that position 63 lies on the OxlT translocation pathway . Our conclusion is strengthened by the finding that position 63, along with most other positions relevant to TM2 function, is found on a helical face that can be cross-linked to the pathway-facing surface of TM11 (Fu, D., Sarker, R . I., Bolton, E., and Maloney, P . C . (2001) J . Biol . Chem . 276, 8753-8760). J Feline Med Surg, 1999 Mar, 1(1), 23 - 9 Disseminated Mycobacterium genavense infection in a FIV-positive cat; Hughes MS et al.; An 8-year-old FIV-positive Australian cat was presented with coughing, periocular alopecia, pyrexia and inappetence . Skin scrapings demonstrated Demodex cati mites . Antibiotics were administered and it was treated successfully for periocular demodectic mange, but the cat continued to exhibit respiratory signs and lose weight . Further investigation revealed an ascarid infection and active chronic inflammation of undetected cause affecting the lower airways . Repetitive treatment with pyrantel failed to eradicate the ascarid infection . The cat became cachectic and developed moist ulcerative dermatitis of the neck, severe non-regenerative anaemia, leucopenia and thrombocytopenia . Necropsy and histopathology revealed mycobacteriosis affecting skin, lungs, spleen, lymph nodes, liver and kidney . Attempted culture of frozen tissues at a mycobacteria reference laboratory was unsuccessful . Paraffin-embedded, formalin-fixed tissue was retrieved and examined using PCR to amplify part of the 16S rRNA gene . A diagnosis of disseminated Mycobacterium genavense infection was made based on the presence of acid fast bacteria in many tissues and partial sequence of the 16S rRNA gene . Although M genavense has been identified previously as a cause of disseminated disease in AIDS patients, this is the first report of infection in a cat . It was suspected that the demodecosis, recurrent ascarid infections and disseminated M genavense infection resulted from an immune deficiency syndrome consequent to longstanding FIV infection . Mol Microbiol, 2002 Mar, 43(5), 1139 - 50 The C-terminus of IcmT is essential for pore formation and for intracellular trafficking of Legionella pneumophila within Acanthamoeba polyphaga; Molmeret M et al.; We have shown previously that the five rib (release of intracellular bacteria) mutants of Legionella pneumophila are competent for intracellular replication but defective in pore formation-mediated cytolysis and egress from protozoan and mammalian cells . The rib phenotype results from a point mutation (deletion) DeltaG544 in icmT that is predicted to result in the expression of a protein truncated by 32 amino acids from the C-terminus . In contrast to the rib mutants that are capable of intracellular replication, an icmT null mutant was completely defective in intracellular replication within mammalian and protozoan cells, in addition to its defect in pore formation-mediated cytolysis . The icmT wild-type allele complemented the icmT null mutant for both defects of intracellular replication and pore formation-mediated cytolysis and egress from mammalian cells . In contrast, the icmTDeltaG544 allele complemented the icmT null mutant for intracellular growth, but not for the pore-forming activity . Consistent with their defect in pore formation-mediated cytotoxicity in vitro, both mutants failed to cause pulmonary inflammation in A/J mice . Interestingly, the rib mutant was severely defective in intracellular growth within Acanthamoeba polyphaga . Confocal laser scanning and electron microscopy confirmed that the rib mutant and the icmT null mutant were severely and completely defective, respectively, in intracellular growth in A . polyphaga, and the respective defects correlated with fusion of the bacterial phagosomes to lysosomes . Taken together, the data showed that the C-terminus domain of IcmT is essential for the pore-forming activity and is required for intracellular trafficking and replication within A . polyphaga, but not within mammalian cells. Mol Microbiol, 2002 Mar, 43(5), 1115 - 27 The Sinorhizobium meliloti stringent response affects multiple aspects of symbiosis; Wells DH et al.; Sinorhizobium meliloti and host legumes enter into a nitrogen-fixing, symbiotic relationship triggered by an exchange of signals between bacteria and plant . S . meliloti produces Nod factor, which elicits the formation of nodules on plant roots, and succinoglycan, an exopolysaccharide that allows for bacterial invasion and colonization of the host . The biosynthesis of these molecules is well defined, but the specific regulation of these compounds is not completely understood . Bacteria control complex regulatory networks by the production of ppGpp, the effector molecule of the stringent response, which induces physiological change in response to adverse growth conditions and can also control bacterial development and virulence . Through detailed analysis of an S . meliloti mutant incapable of producing ppGpp, we show that the stringent response is required for nodule formation and regulates the production of succinoglycan . Although it remains unknown whether these phenotypes are connected, we have isolated suppressor strains that restore both defects and potentially identify key downstream regulatory genes . These results indicate that the S . meliloti stringent response has roles in both succinoglycan production and nodule formation and, more importantly, that control of bacterial physiology in response to the plant and surrounding environment is critical to the establishment of a successful symbiosis. Nucleic Acids Res, 2002 Apr 1, 30(7), 1427 - 64 Comparative genomics and evolution of proteins involved in RNA metabolism; Anantharaman V et al.; RNA metabolism, broadly defined as the compendium of all processes that involve RNA, including transcription, processing and modification of transcripts, translation, RNA degradation and its regulation, is the central and most evolutionarily conserved part of cell physiology . A comprehensive, genome-wide census of all enzymatic and non-enzymatic protein domains involved in RNA metabolism was conducted by using sequence profile analysis and structural comparisons . Proteins related to RNA metabolism comprise from 3 to 11% of the complete protein repertoire in bacteria, archaea and eukaryotes, with the greatest fraction seen in parasitic bacteria with small genomes . Approximately one-half of protein domains involved in RNA metabolism are present in most, if not all, species from all three primary kingdoms and are traceable to the last universal common ancestor (LUCA) . The principal features of LUCA's RNA metabolism system were reconstructed by parsimony-based evolutionary analysis of all relevant groups of orthologous proteins . This reconstruction shows that LUCA possessed not only the basal translation system, but also the principal forms of RNA modification, such as methylation, pseudouridylation and thiouridylation, as well as simple mechanisms for polyadenylation and RNA degradation . Some of these ancient domains form paralogous groups whose evolution can be traced back in time beyond LUCA, towards low-specificity proteins, which probably functioned as cofactors for ribozymes within the RNA world framework . The main lineage-specific innovations of RNA metabolism systems were identified . The most notable phase of innovation in RNA metabolism coincides with the advent of eukaryotes and was brought about by the merge of the archaeal and bacterial systems via mitochondrial endosymbiosis, but also involved emergence of several new, eukaryote-specific RNA-binding domains . Subsequent, vast expansions of these domains mark the origin of alternative splicing in animals and probably in plants . In addition to the reconstruction of the evolutionary history of RNA metabolism, this analysis produced numerous functional predictions, e.g . of previously undetected enzymes of RNA modification. Appl Environ Microbiol, 2002 Apr, 68(4), 2044 - 8 Bradyrhizobia from wild Phaseolus, Desmodium, and Macroptilium species in northern Mexico; Parker MA; rRNA genetic markers were analyzed in 97 isolates of nodule bacteria from six legume species in Chihuahua, Mexico . The most common genotypes were widely shared across host species and had 16S rRNA sequences identical to those of strains from an eastern North American legume (Amphicarpaea) that are closely related to Bradyrhizobium elkanii. Appl Environ Microbiol, 2002 Apr, 68(4), 1932 - 7 Desulfovibrio sp . genes involved in the respiration of sulfate during metabolism of hydrogen and lactate; Steger JL et al.; To develop a better understanding of respiration by sulfate-reducing bacteria, we examined transcriptional control of respiratory genes during growth with lactate or hydrogen as an electron donor . RNA extracts of Desulfovibrio desulfuricans subsp . aestuarii were analyzed by using random arbitrarily primed PCR . RNA was reverse transcribed under low-stringency conditions with a set of random primers, and candidate cDNAs were cloned, sequenced, and characterized by BLAST analysis . Putative differentially expressed transcripts were confirmed by Northern blot analysis . Interestingly, dissimilatory bisulfite reductase was upregulated in the presence of hydrogen . To link these transcriptional changes to the physiology of sulfate-reducing bacteria, sulfide was measured during growth of several strains of Desulfovibrio on hydrogen or lactate, and this revealed that hydrogen-grown cells produced more sulfide per unit of cell mass than lactate-grown cells . Transcription of other redox proteins was characterized by Northern blotting to determine whether or not they were also transcribed to higher levels in hydrogen-grown cells . Growth on lactate produced greater transcription of {NiFe} hydrogenase . H(2)-grown cells transcribed the adenylylsulfate reductase b subunit and HmcA to higher levels . The results we describe here provide new insight into the continuing debate over how Desulfovibrio species utilize redox components to generate membrane potential and to channel electrons to sulfate, the final electron acceptor. Appl Environ Microbiol, 2002 Apr, 68(4), 1690 - 6 NAD(P)H:flavin mononucleotide oxidoreductase inactivation during 2,4,6-trinitrotoluene reduction; Riefler RG et al.; Bacteria readily transform 2,4,6-trinitrotoluene (TNT), a contaminant frequently found at military bases and munitions production facilities, by reduction of the nitro group substituents . In this work, the kinetics of nitroreduction were investigated by using a model nitroreductase, NAD(P)H:flavin mononucleotide (FMN) oxidoreductase . Under mediation by NAD(P)H:FMN oxidoreductase, TNT rapidly reacted with NADH to form 2-hydroxylamino-4,6-dinitrotoluene and 4-hydroxylamino-2,6-dinitrotoluene, whereas 2-amino-4,6-dinitrotoluene and 4-amino-2,6-dinitrotoluene were not produced . Progressive loss of activity was observed during TNT reduction, indicating inactivation of the enzyme during transformation . It is likely that a nitrosodinitrotoluene intermediate reacted with the NAD(P)H:FMN oxidoreductase, leading to enzyme inactivation . A half-maximum constant with respect to NADH, K(N), of 394 microM was measured, indicating possible NADH limitation under typical cellular conditions . A mathematical model that describes the inactivation process and NADH limitation provided a good fit to TNT reduction profiles . This work represents the first step in developing a comprehensive enzyme level understanding of nitroarene biotransformation. Appl Environ Microbiol, 2002 Apr, 68(4), 1665 - 73 Photosynthetic apparatus in Roseateles depolymerans 61A is transcriptionally induced by carbon limitation; Suyama T et al.; Production of a photosynthetic apparatus in Roseateles depolymerans 61A, a recently discovered freshwater beta-Proteobacterium showing characteristics of aerobic phototrophic bacteria, was observed when the cells were subjected to a sudden decrease in carbon sources (e.g., when cells grown with 0.1 to 0.4% Casamino Acids were diluted or transferred into medium containing <or=0.04% Casamino Acids) . Accumulation of bacteriochlorophyll (BChl) a was observed in the presence of oxygen and was enhanced under semiaerobic conditions (2% oxygen) but was reduced in the presence of light . Similarly to what has been reported regarding some aerobic phototrophic bacteria belonging to the alpha subclass of the Proteobacteria, viability of the cells in the carbon source-free medium was prolonged under aerobic-light (10 W m(-2)) conditions, possibly due to photosynthetic energy conversion, but was not prolonged under aerobic-dark conditions . The puf operon, which encodes most of the apoproteins of light-harvesting and reaction center complexes, was sequenced, and the effect of changes in Casamino Acids concentrations, oxygen, and light on its expression was estimated by the accumulation of its mRNA . The expression of the puf operon was induced by the decrease in carbon sources, similarly to what was observed for the accumulation of BChl a under aerobic and semiaerobic conditions (>or=0.2% O(2)), and was reduced in the presence of light . Transcription of the R . depolymerans puf operon is considered to be controlled by changes in carbon nutrients in addition to oxygen tension and light intensity. Fetal Diagn Ther, 2002 May-Jun, 17(3), 167 - 72 Soluble intercellular adhesion molecule 1 in umbilical cord serum: potential for the diagnosis of neonatal infections; Darai E et al.; OBJECTIVE: To evaluate the diagnostic relevance to neonatal infections of the soluble intercellular adhesion molecule 1 (sICAM-1) cord serum level . METHODS: The case-control study included 66 term newborn infants with and without risk factors for neonatal infections . Cord blood serum determinations of white blood cell count, C-reactive protein, fibrinogen, and sICAM-1 were systematically performed associated with bacterial cultures from placenta, ears, and gastric fluids . RESULTS: 6 of 33 infants (18.2%) with risk factors were infected, and 13 (39.4%) were colonized . Two infants included in the group without infection risk factors (n = 33) were colonized . No difference in sICAM-1 cord serum levels was found according to the presence of premature rupture of membrane, fetal tachycardia >160 bpm, meconial amniotic fluid, and duration of labour >10 h . No difference in sICAM-1 was noted between infected and non-infected infants . The cord serum levels of sICAM-1 were significantly higher in infants after forceps extraction (p = 0.01) . A correlation was observed between sICAM-1 and C-reactive protein cord serum levels (p = 0.004, r = 0.371) and between sICAM-1 level and neutrophil count (p = 0.01, r = 0.489) . CONCLUSIONS: Our results suggest that cord serum sICAM-1 determinations have no diagnostic relevance to neonatal infection . The increase of sICAM-1 cord serum levels in infants after forceps extraction suggests its potential to evaluate cerebral trauma or hypoxia . Cell Physiol Biochem, 2002, 12(1), 19 - 30 Simvastatin inhibits malignant transformation following expression of the Ha-ras oncogene in NIH 3T3 fibroblasts; Furst J et al.; In previous studies we have shown that the expression of the transforming Ha-ras oncogene in NIH 3T3 fibroblasts stimulates cellular calcium entry, which triggers oscillatory calcium induced calcium release from internal stores . The intracellular calcium oscillations lead to cytoskeletal remodeling by actin stress fiber depolymerization and activation of the Na(+)/H(+) exchanger thus mediating cell swelling and intracellular alkalosis, both important mitogenic signals . This is evidenced by abrogation of Ha-ras induced growth factor independent cell proliferation by interference with any of these events, i.e . by inhibition of cellular calcium entry or inhibition of the Na(+)/H(+) exchanger.As shown in this study, simvastatin, an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, the key enzyme for cholesterol biosynthesis, is able to prevent these events following the expression of the transforming Ha-ras oncogene . We show, that simvastatin inhibits farnesylation dependent membrane translocation of a CAAX motive bearing yellow fluorescent protein and suppresses Ha-ras stimulated cellular calcium influx, which can be identified as capacitative calcium entry . In addition simvastatin is able to block regulatory volume decrease channels and to suppress the cytoskeletal remodeling, intracellular alkalinization, increase in cell volume and growth factor independent cell proliferation induced by the oncogene.Thus simvastatin is able to prevent crucial cellular events following expression of the transforming Ha-ras oncogene . Acta Crystallogr D Biol Crystallogr, 2002 Apr, 58(Pt 4), 653 - 9 Epub 2002 Mar 22. Structure of an endoglucanase from termite, Nasutitermes takasagoensis; Khademi S et al.; Contrary to conventional wisdom, it has been shown recently that termites do not necessarily depend on symbiotic bacteria to process cellulose . They secrete their own cellulases, mainly endo-beta-1,4-glucanase and beta-1,4-glucosidase . Here, the first structure of an endogenous endoglucanase from the higher termite Nasutitermes takasagoensis (NtEgl) is reported at 1.40 A resolution . NtEgl has the general folding of an (alpha/alpha)(6) barrel, which is a common folding pattern for glycosyl hydrolase family 9 . Three-dimensional structural analysis shows that the conserved Glu412 is the catalytic acid/base residue and the conserved Asp54 or Asp57 is the base . The enzyme has a Ca(2+)-binding site near its substrate-binding cleft . Comparison between the structure of the Ca(2+)-free enzyme produced by reducing the pH of the soaked crystal from 5.6 (the pH of optimum enzyme activity) to 2.5 with that of the Ca(2+)-bound enzyme did not show significant differences in the locations of the C(alpha) atoms . The main differences are in the conformation of the residue side chains ligating the Ca(2+) ion . The overall structure of NtEgl at pH 6.5 is similar to that at pH 5.6 . The major change observed was in the conformation of the side chain of the catalytic acid/base Glu412, which rotates from a hydrophobic cavity to a relatively hydrophilic environment . This side-chain displacement may decrease the enzyme activity at higher pH. Acta Crystallogr D Biol Crystallogr, 2002 Apr, 58(Pt 4), 585 - 90 Epub 2002 Mar 22. Structure of the photoactive yellow protein reconstituted with caffeic acid at 1.16 A resolution; van Aalten DM et al.; A structural study is described of the photoactive yellow protein (PYP) reconstituted with the chromophore derivative 3,4-dihydroxycinnamic acid . The crystal structure of PYP reconstituted with this chromophore at 1.16 A resolution is reported in space group P6(5) . This is the first high-resolution structure of a photoreceptor containing a modified chromophore . The introduction of an extra hydroxyl group in the native chromophore (i.e . p-coumaric acid) appears to perturb the structure of the hybrid yellow protein only slightly . The chromophore is bound by the protein in two different conformations, separated by a rotation of 180 degrees of the catechol ring . In combination with available spectroscopic data, it is concluded that the caffeic acid chromophore binds to the protein in a strained conformation, which leads to a faster ejection from the chromophore-binding pocket upon pB formation. J Biol Chem, 2002 May 31, 277(22), 19709 - 19 Epub 2002 Mar 25. Lactotetraosylceramide, a novel glycosphingolipid receptor for Helicobacter pylori, present in human gastric epithelium; Teneberg S et al.; The binding of Helicobacter pylori to glycosphingolipids was examined by binding of (35)S-labeled bacteria to glycosphingolipids on thin-layer chromatograms . In addition to previously reported binding specificities, a selective binding to a non-acid tetraglycosylceramide of human meconium was found . This H . pylori binding glycosphingolipid was isolated and, on the basis of mass spectrometry, proton NMR spectroscopy, and degradation studies, were identified as Galbeta3GlcNAcbeta3Galbeta4Glcbeta1Cer (lactotetraosylceramide) . When using non-acid glycosphingolipid preparations from human gastric epithelial cells, an identical binding of H . pylori to the tetraglycosylceramide interval was obtained in one of seven samples . Evidence for the presence of lactotetraosylceramide in the binding-active interval was obtained by proton NMR spectroscopy of intact glycosphingolipids and by gas chromatography-electron ionization mass spectrometry of permethylated tetrasaccharides obtained by ceramide glycanase hydrolysis . The lactotetraosylceramide binding property was detected in 65 of 74 H . pylori isolates (88%) . Binding of H . pylori to lactotetraosylceramide on thin-layer chromatograms was inhibited by preincubation with lactotetraose but not with lactose . Removal of the terminal galactose of lactotetraosylceramide by galactosidase hydrolysis abolished the binding as did hydrazinolysis of the acetamido group of the N-acetylglucosamine . Therefore, Galbeta3GlcNAc is an essential part of the binding epitope. J Bacteriol, 2002 Apr, 184(8), 2072 - 80 Inferring genome trees by using a filter to eliminate phylogenetically discordant sequences and a distance matrix based on mean normalized BLASTP scores; Clarke GD et al.; Darwin's paradigm holds that the diversity of present-day organisms has arisen via a process of genetic descent with modification, as on a bifurcating tree . Evidence is accumulating that genes are sometimes transferred not along lineages but rather across lineages . To the extent that this is so, Darwin's paradigm can apply only imperfectly to genomes, potentially complicating or perhaps undermining attempts to reconstruct historical relationships among genomes (i.e., a genome tree) . Whether most genes in a genome have arisen via treelike (vertical) descent or by lateral transfer across lineages can be tested if enough complete genome sequences are used . We define a phylogenetically discordant sequence (PDS) as an open reading frame (ORF) that exhibits patterns of similarity relationships statistically distinguishable from those of most other ORFs in the same genome . PDSs represent between 6.0 and 16.8% (mean, 10.8%) of the analyzable ORFs in the genomes of 28 bacteria, eight archaea, and one eukaryote (Saccharomyces cerevisiae) . In this study we developed and assessed a distance-based approach, based on mean pairwise sequence similarity, for generating genome trees . Exclusion of PDSs improved bootstrap support for basal nodes but altered few topological features, indicating that there is little systematic bias among PDSs . Many but not all features of the genome tree from which PDSs were excluded are consistent with the 16S rRNA tree. J Dairy Sci, 2002 Feb, 85(2), 434 - 44 Corn silage management: effects of maturity, inoculation, and mechanical processing on pack density and aerobic stability; Johnson LM et al.; Three experiments were conducted to evaluate the effects of inoculation, maturity, and mechanical processing of corn silage on aerobic stability and pack density . Corn silage was stored in 20-L mini silos for the three aerobic stability experiments . Corn silage was stored in 80-L mini silos for the three pack-density experiments . The wet pack density of corn silage tended to decrease as maturity advanced in all of the pack-density experiments, and processed corn silage had a greater wet pack density compared with unprocessed corn silage in two of the three 20-L mini silo experiments . Aerobic stability, measured as the number of hours to reach 1.7 degrees C above ambient, was greater for processed corn silage in two of the three 20-L mini silo experiments, and was greater for inoculated corn silage across the three 20-L mini silo experiments . Inoculation of corn silage with lactic acid producing bacteria tended to improve aerobic stability of corn silage more than maturity and mechanical processing. Trends Microbiol, 2002 Apr, 10(4), 169 - 73 The Schaechter-Bentzon-Maaløe experiment and the analysis of cell cycle events in eukaryotic cells; Cooper S; The Schaechter-Bentzon-Maaloe (SBM) experiment, performed more than 40 years ago, provides an important lesson for the analysis of the eukaryotic cell cycle . Before this experiment, temperature shifts had been used to synchronize bacteria and determine the pattern of DNA synthesis during the bacterial division cycle . These experiments indicated that DNA replication occurred during a fraction of the division cycle with gaps before and after DNA synthesis, a pattern similar to the eukaryotic division cycle . The SBM experiment studied DNA replication during the division cycle by labeling an unperturbed culture with a short pulse of tritiated thymidine . All cells were found to be labeled, indicating that unperturbed cells synthesize DNA throughout the division cycle . Thus, the SBM experiment was a control experiment demonstrating that artifacts can be introduced by synchronization methods . The idea of an control experiment under unperturbed conditions is proposed for the analysis of data on cell-cycle-specific gene expression in yeast and mammalian cells. Microbiol Res, 2002, 157(1), 25 - 8 Isolation and characterization of the major nod factor of Bradyrhizobium japonicum strain 532C; Soulemanov A et al.; Bradyrhizobium japonicum 532C nodulates soybean effectively under cool Canadian spring conditions and is used in Canadian commercial inoculants . The major lipo-chitooligosaccharide (LCO), bacteria-to-plant signal was characterized by HPLC, FAB-mass spectroscopy MALDI-TOF mass spectroscopy and revealed to be LCO Nod Bj-V (C18:1, MeFuc) . This LCO is produced by type I strains of B . japonicum and is therefore unlikely to account for this strains superior ability to nodulate soybean under Canadian conditions . We also found that use of yeast extract mannitol medium gave similar results to that of Bergerson minimal medium. Dig Dis Sci, 2002 Mar, 47(3), 489 - 94 The glucose breath test: a diagnostic test for small bowel stricture(s) in Crohn's disease; Mishkin D et al.; The aim of this study was to determine whether an indirect noninvasive indicator of proximal bacterial overgrowth, the glucose breath test, was of diagnostic value in inflammatory bowel disease . Twenty four of 71 Crohn's disease patients tested had a positive glucose breath test . No statistical conclusions could be drawn between the Crohn's disease activity index and glucose breath test status . Of patients with radiologic evidence of small bowel stricture(s), 96.0% had a positive glucose breath test, while only one of 46 negative glucose breath test patients had a stricture . The positive and negative predictive values for a positive glucose breath test as an indicator of stricture formation were 96.0% and 97.8%, respectively . This correlation was not altered in Crohn's disease patients with fistulae or status postresection of the terminal ileum . The data in ulcerative colitis were nondiagnostic . In conclusion, the glucose breath test appears to be an accurate noninvasive inexpensive diagnostic test for small bowel stricture(s) and secondary bacterial overgrowth in Crohn's disease. Gastroenterol Clin Biol, 2001 Dec, 25(12), 1084 - 9 {CagA status and virulence of Helicobacter pylori strains . Results of a French multicentric prospective study}; Bommelaer G et al.; Previous experimental and epidemiological studies with few patients suggested that the presence of the cagA gene was a virulence factor for Helicobacter pylori (H . pylori) . AIM: To establish in this large epidemiological cohort study the relationship between the histological virulence of H . pylori infection and the cagA status of the bacteria . METHODS: This prospective cohort study (6 month follow-up) was conducted on adult patients undergoing endoscopy for upper gastrointestinal symptoms . The cagA status of H . pylori-positive patients was established using the polymerase chain reaction (PCR) method on an antral biopsy . A score of histological virulence (inflammation, activity) was recorded on the basis of the Sydney system (on antral, angular and fundic biopsies) . Eradication treatment given was not imposed and a clinical follow-up was performed at 3 and 6 months . H . pylori eradication was verified by a 13C urea breath test at 3 months . RESULTS: Four hundred and twenty two centers recruited 652 patients (mean age: 51 +/- 15 years, 55% female) . Upper GI endoscopy was abnormal in 80% of the patients of whom 68% had a gastritis aspect; 38% were infected by H . pylori, and among them 51% were cagA-positive . The histological virulence scores associated with the cagA-positive strains were significantly higher than those associated with the cagA-negative strains, globally (P = 0.0035), in the antrum (P = 0.0063), and in the angulus (P = 0.046), but not in the fundus (P = 0.05) . The cagA status was correlated neither with the symptom severity at inclusion and at 6 months (P > 0.05), nor with the H . pylori eradication rate at 3 months (75% in cagA-positive and 70% in cagA-negative strains, P = 0.52) . CONCLUSION: This study on a large cohort of patients confirms the greater histological virulence of H . pylori cagA-positive strains . However, this virulence was not associated with more severe symptoms nor with an increase in resistance to H . pylori eradication treatment. Curr Microbiol, 2002 Apr, 44(4), 241 - 5 Three-piece-ligation PCR and application in disruption of chlorophyll synthesis genes in Synechocystis sp . PCC 6803; Kong R et al.; Linear DNA, consisting of a drug-resistance marker and long flanking sequences, was synthesized by one-step polymerase chain reaction after a three-piece ligating reaction . Chlorophyll synthesis genes, chlH and chlL in Synechocystis sp . PCC 6803, were replaced by a kanamycin-resistance marker through double recombinations with flanking homology regions . Under LAHG conditions, the chlL but not chlH mutant stopped chlorophyll synthesis, while both synthesized chlorophyll in the light. Plant Cell, 2002 Mar, 14(3), 559 - 74 Expression profile matrix of Arabidopsis transcription factor genes suggests their putative functions in response to environmental stresses; Chen W et al.; Numerous studies have shown that transcription factors are important in regulating plant responses to environmental stress . However, specific functions for most of the genes encoding transcription factors are unclear . In this study, we used mRNA profiles generated from microarray experiments to deduce the functions of genes encoding known and putative Arabidopsis transcription factors . The mRNA levels of 402 distinct transcription factor genes were examined at different developmental stages and under various stress conditions . Transcription factors potentially controlling downstream gene expression in stress signal transduction pathways were identified by observed activation and repression of the genes after certain stress treatments . The mRNA levels of a number of previously characterized transcription factor genes were changed significantly in connection with other regulatory pathways, suggesting their multifunctional nature . The expression of 74 transcription factor genes responsive to bacterial pathogen infection was reduced or abolished in mutants that have defects in salicylic acid, jasmonic acid, or ethylene signaling . This observation indicates that the regulation of these genes is mediated at least partly by these plant hormones and suggests that the transcription factor genes are involved in the regulation of additional downstream responses mediated by these hormones . Among the 43 transcription factor genes that are induced during senescence, 28 of them also are induced by stress treatment, suggesting extensive overlap responses to these stresses . Statistical analysis of the promoter regions of the genes responsive to cold stress indicated unambiguous enrichment of known conserved transcription factor binding sites for the responses . A highly conserved novel promoter motif was identified in genes responding to a broad set of pathogen infection treatments . This observation strongly suggests that the corresponding transcription factors play general and crucial roles in the coordinated regulation of these specific regulons . Although further validation is needed, these correlative results provide a vast amount of information that can guide hypothesis-driven research to elucidate the molecular mechanisms involved in transcriptional regulation and signaling networks in plants. Mol Cell Biol, 2002 Apr, 22(8), 2842 - 52 Requirement of TRAP/mediator for both activator-independent and activator-dependent transcription in conjunction with TFIID-associated TAF(II)s; Baek HJ et al.; The multiprotein human TRAP/Mediator complex, which is phylogenetically related to the yeast SRB/Mediator coactivator, facilitates activation through a wide variety of transcriptional activators . However, it remains unclear how TRAP/Mediator functions in the context of other coactivators . Here we have identified a previously uncharacterized integral subunit (TRAP25) of the complex that is apparently metazoan specific . An antibody that is specific for TRAP25 allowed quantitative immunodepletion of essentially all TRAP/Mediator components from HeLa nuclear extract, without detectably affecting levels of RNA polymerase II and corresponding general transcription factors . Surprisingly, the TRAP/Mediator-depleted nuclear extract displayed severely reduced levels of both basal and activator-dependent transcription from DNA templates . Both activities were efficiently restored upon readdition of purified TRAP/Mediator . Moreover, restoration of basal and activator-dependent transcription to extracts that were simultaneously depleted of TRAP/Mediator and TFIID (TBP plus the major TAF(II)s) required addition of both TBP and associated TAF(II)s, as well as TRAP/Mediator . These observations indicate that TAF(II)s and Mediator are jointly required for both basal and activated transcription in the context of a more physiological complement of nuclear proteins . We propose a close mechanistic linkage between these components that most likely operates at the level of combined effects on the general transcription machinery and, in addition, a direct role for Mediator in relaying activation signals to this machinery. Microbes Infect, 2002 Mar, 4(3), 333 - 40 Proteins in the chlamydial inclusion membrane; Rockey DD et al.; The chlamydiae are obligate intracellular pathogens that occupy a nonacidified vacuole (the inclusion) during their entire developmental cycle . Several proteins have recently been identified that are localized to the inclusion membrane . The following is a discussion of how inclusion membrane proteins might participate in the chlamydial developmental process. Cell, 2002 Feb 22, 108(4), 557 - 72 Ribosome structure and the mechanism of translation; Ramakrishnan V; The publication of crystal structures of the 50S and 30S ribosomal subunits and the intact 70S ribosome is revolutionizing our understanding of protein synthesis . This review is an attempt to correlate the structures with biochemical and genetic data to identify the gaps and limits in our current knowledge of the mechanisms involved in translation. Genome, 2002 Feb, 45(1), 116 - 24 A DNA mismatch repair gene links to the Ph2 locus in wheat; Dong C et al.; DNA mismatch repair is an essential system for maintaining genetic stability in bacteria and higher eukaryotes . Based on the conserved regions of the bacterial MutS gene and its homologues in yeast and human, a wheat cDNA homologue of MSH6, designated TaMSH7, was isolated by RT-PCR . The deduced amino acid sequence of TaMSH7 shows conserved domains characteristic of other MSH6 genes, with highest similarity to maize MSH7 and Arabidopsis MSH7 . TaMSH7 is expressed in meristem tissue associated with a high level of mitotic and meiotic activity, with maximum expression in the reproductive organs of young flower spikes . TaMSH7 is located on the short arms of chromosomes 3A, 3B, and 3D and has been mapped within barley chromosome 3HS . The copy on 3DS is located within the region deleted in the wheat mutant ph2a, which shows altered recombination frequency in the interspecific hybrids . The relationship between the ph2a mutant and TaMSH7 gene function is discussed. Biochem Cell Biol, 2002, 80(1), 1 - 6 The physiology of lactoferrin; Brock JH; This paper reviews our current knowledge of the structure and function of the iron-binding protein lactoferrin . In particular, it attempts to relate the various proposed physiological functions of lactoferrin to its most characteristic biochemical properties, i.e . its ability to bind iron and its highly basic nature . The extent to which various physiological functions can be considered as definitely established is critically reviewed, and suggestions for future research are proposed. Compend Contin Educ Dent Suppl, 2000, (30), 5 - 11; quiz 65 Preterm birth, osteoporosis, and periodontal disease; Jeffcoat MK; The purpose of this two-part article is to review two major events in the life span of a woman . These include the putative relationship between oral health, pregnancy, and postmenopausal osteoporosis . Current knowledge about risk factors for preterm birth and for osteoporosis are discussed . The newest studies that address the relationship between oral and systemic health are also reviewed. Compend Contin Educ Dent, 2000 Oct, 21(10A), 880 - 6, 888-9; quiz 890 Genuine halitosis, pseudo-halitosis, and halitophobia: classification, diagnosis, and treatment; Yaegaki K et al.; Although tongue brushing and appropriate mouthrinses are both important and basic treatment measures for halitosis, other dental treatments are sometimes required . The treatment of genuine halitosis caused by oral conditions is not complex . In addition to genuine halitosis patients, psychosomatic halitosis patients also visit dental practitioners . Although psychosomatic halitosis is out of the treatment realm of dental practitioners, patients with this condition will still seek help from a dental practitioner . They often only receive treatment for genuine halitosis without referral to a psychological specialist . If these psychosomatic halitosis patients are incorrectly managed, the psychological condition might become worse than before the visit . To avoid the mismanagement of halitosis patients, classifications of halitosis patients have been established . Genuine halitosis was subclassified as physiologic halitosis and pathologic halitosis . Pathologic halitosis was further categorized to oral pathologic halitosis and extraoral pathologic halitosis . Both pseudo-halitosis and halitophobia patients complain of the existence of halitosis, which is not offensive . Pseudo-halitosis cannot be treated by dental practitioners, and halitophobia patients must be referred to psychological specialists . Clinicians need to examine the psychological condition of halitosis patients at the initial patient visit . A questionnaire prepared for the clinic at the University of British Columbia was found to be advantageous for this purpose. J Biol Chem, 2002 May 24, 277(21), 19080 - 6 Epub 2002 Mar 20. Characterization and expression of L-amino acid oxidase of mouse milk; Sun Y et al.; l-Amino acid oxidase (LAO) was purified from mouse milk . LAO reacted with l-amino acids in an apparent order of Phe > Met, Tyr > Cys, Leu > His other 11 amino acids tested and produced H(2)O(2) in a dose- and time-dependent manner . LAO in milk had a molecular mass of about 113 kDa and was converted to a 60-kDa protein by SDS-PAGE . LAO consisted of two subunits . The N- and C-terminal amino acid sequence determination followed by cDNA cloning showed that the 60-kDa protein consisted of 497 amino acids . LAO mRNA spanned about 2.0 kb, and its expression was found only in the mammary epithelial cells . Glucocorticoid was essential for LAO gene expression . Thus, the LAO gene is expressed acutely upon the onset of milk synthesis . LAO mRNA increased 1 day before parturition, peaked during early to mid-lactation, and decreased at the end of lactation . This is the first demonstration showing that LAO is present in milk . Mastitis is caused by an intramammary bacterial infection . As mouse milk produced H(2)O(2) using endogenous free amino acids, we suggest that LAO, together with free amino acids, is responsible for killing bacteria in the mammary gland. Biol Reprod, 2002 Apr, 66(4), 1193 - 202 Analysis of the roles of RGD-binding integrins, alpha(4)/alpha(9) integrins, alpha(6) integrins, and CD9 in the interaction of the fertilin beta (ADAM2) disintegrin domain with the mouse egg membrane; Zhu X et al.; Fertilin beta (also known as ADAM2), a mammalian sperm protein that mediates gamete cell adhesion during fertilization, is a member of the ADAM protein family whose members have disintegrin domains with homology to integrin ligands found in snake venoms . Fertilin beta utilizes an ECD sequence within its disintegrin domain to interact with the egg plasma membrane; the Asp is especially critical . Based on what is known about different integrin subfamilies and their ligands, we sought to characterize fertilin beta binding sites on mouse eggs, focusing on integrin subfamilies that recognize short peptide sequences that include an Asp residue: the alpha(5)/alpha(8)/alpha(v)/alpha(IIb) or RGD-binding subfamily (alpha(5)beta(1), alpha(8)beta(1), alpha(V)beta(1), alpha(V)beta(3), alpha(V)beta(5), alpha(V)beta(6), alpha(V)beta(8), and alpha(IIb)beta(3)) and the alpha(4)/alpha(9) subfamily (alpha(4)beta(1), alpha(9)beta(1), and alpha(4)beta(7)) . We tested peptide sequences known to perturb interactions mediated by these integrins in two different assays for fertilin beta binding . Peptides with the sequence MLDG, which perturb alpha(4)/alpha(9) integrin-mediated interactions, significantly inhibit fertilin beta binding to eggs, which suggests a role for a member of this integrin subfamily as a fertilin beta receptor . RGD peptides, which perturb alpha(5)/alpha(8)/alpha(v)/alpha(IIb) integrin-mediated interactions, have partial inhibitory activity . The anti-alpha(6) antibody GoH3 has little or no inhibitory activity . An antibody to the integrin-associated tetraspanin protein CD9 inhibits the binding of a multivalent presentation of fertilin beta (immobilized on beads) but not soluble fertilin beta, which we speculate has implications for the role of CD9 in the strengthening of fertilin beta-mediated cell adhesion but not in initial ligand binding. Trends Plant Sci, 2002 Mar, 7(3), 103 - 5 Rings and networks: the amazing complexity of FtsZ in chloroplasts; Reski R; Bacteria have proteins that can form filaments and rings, and these are thought to be the evolutionary progenitors of actin and tubulin . Plant homologues of the most intensively studied bacterial FtsZ protein are nuclear-encoded by a small gene family, are plastid-bound and participate in the plastid division process . The hypothesis is put forward that FtsZ and other proteins form a filamentous network in plastids, a plastoskeleton, which keeps these organelles in shape and helps them to divide. Eur J Neurosci, 2002 Mar, 15(5), 798 - 806 Selective targeting of zebrafish olfactory receptor neurons by the endogenous OMP promoter; Celik A et al.; The olfactory nervous system of fish, in particular zebrafish, has become a valid model for that of higher vertebrates . However, no genetic markers for olfactory specific cell types, e.g . the olfactory receptor neurons, have been established in this species . Olfactory marker protein (OMP) is a reliable marker for olfactory receptor neurons in several other vertebrates . We have cloned zOMP, the zebrafish homologue of olfactory marker protein . During development, zOMP is expressed exclusively in the olfactory placode, presumably in olfactory receptor neurons, as shown by in situ hybridization . In the adult nasal epithelium zOMP is found restricted to the sensory region . zOMP appears to be a single gene, without close family members . The 5'-flanking region lacks most of the expected regulatory sequence motifs, both general and cell type-specific ones . Nevertheless, it drives reporter gene expression strongly and specifically in olfactory receptor neurons during the whole developmental period examined . Thus the zOMP promoter constitutes a powerful tool which should be useful to selectively introduce a wide variety of genetic modifications into olfactory receptor neurons. Biochem Biophys Res Commun, 2002 Mar 29, 292(2), 293 - 9 The involvement of molybdenum in life; Williams RJ et al.; Quite extraordinarily molybdenum is an essential element in life for the uptake of nitrogen from both nitrogen gas and nitrate, yet it is a relatively rare heavy trace element . It also functions in a few extremely important oxygen-atom transfer reactions at low redox potential . This review poses the question "Why does life depend upon molybdenum?" The answer has to be based upon the availability of the element and on chemical superiority in carrying out the essential tasks . We illustrate here the peculiarities of molybdenum chemistry and how they have become part of certain enzymes . The uptake and incorporation of molybdenum are dependent on its availability, selective pumps, and carriers (chaperones), but 4.5 x 10(9) years ago molybdenum was not available when both tungsten and vanadium or even iron were possibly used in its place . While these possibilities are explored, they leave many unanswered questions concerning the selection today of molybdenum . (c)2002 Elsevier Science (USA). Lancet Oncol, 2001 Aug, 2(8), 483 - 90 Repair of DNA interstrand crosslinks: molecular mechanisms and clinical relevance; McHugh PJ et al.; Drugs that produce DNA interstrand crosslinks (ICLs), between the two complementary strands of the double helix, have an important role in chemotherapy regimens for cancer . Novel crosslinking agents, and targeting strategies involving DNA crosslinking agents, continue to be developed . The ability of cells to repair DNA ICLs is a critical determinant of sensitivity, and recent dinical studies indicate that DNA repair capacity is strongly implicated in both inherent tumour sensitivity and acquired drug resistance . A detailed understanding of the cellular mechanisms that act to eliminate these critical DNA lesions is clearly important . DNA ICLs present a complex challenge to DNA repair mechanisms because of the involvement of both DNA strands . It is now clear that cells from bacteria and yeast to mammals eliminate interstrand ICLs through the coordinated action of several DNA repair pathways . Recently, a model of ICL repair has been proposed, in which mammalian cells use novel excision repair reactions (requiring the XPF and ERCC1 proteins) to uncouple the crosslink . This is followed by a homologous recombination step to provide the genetic information needed to complete repair . This new knowledge may permit the development of screens for tumour response to crosslinking agents, and should also aid the design of more effective crosslinking agents that evade DNA repair . In addition, the proteins mediating the repair reactions represent potential targets for therapeutic intervention. Lancet Oncol, 2002 Jan, 3(1), 17 - 26 Oncolytic biotherapy: a novel therapeutic plafform; Hawkins LK et al.; There is a clear need for new, selective, cancer treatments that do not cause the cross-resistance which occurs with currently available chemotherapeutic agents . Gene therapy is a promising approach, but to date, it has shown limited effectiveness in clinical trials because of insufficient gene transduction . Many investigators are now revisiting the 'old' idea of using tumour-specific, replication-selective viruses or bacteria to treat cancer . These agents can be directly oncolytic, but can also be used to simultaneously express therapeutic genes in target cells or induce tumour-specific, cell-mediated immunity . We discuss the promise of this rapidly evolving field and examine the potential barriers to its success. Intensive Care Med, 2002 Mar, 28(3), 272 - 7 Epub 2002 Feb 08. Combination of venoarterial PCO2 difference with arteriovenous O2 content difference to detect anaerobic metabolism in patients; Mekontso-Dessap A et al.; OBJECTIVE: Under conditions of tissue hypoxia total CO2 production (VCO2) should be less reduced than O2 consumption (VO2) since an anaerobic CO2 production should occur . Thus the VCO(2)/VO(2) ratio, and hence the venoarterial CO2 tension difference/arteriovenous O2 content difference ratio (DeltaPCO2/C(a-v)O2), should increase . We tested the value of the DeltaPCO2/C(a-v)O2 ratio in detecting the presence of global anaerobic metabolism as defined by an increase in arterial lactate level above 2 mmol/l (Lac+) . DESIGN AND SETTING: Retrospective study over a 17-month period in medical intensive care unit of a university hospital . PATIENTS: We obtained 148 sets of measurements in 89 critically ill patients monitored by a pulmonary artery catheter . RESULTS: The DeltaPCO2/C(a-v)O2 ratio was higher in those with increased ( n=73) than in the normolactatemic group (2.0+/-0.9 vs . 1.1+/-0.6, p<0.0001) . Among all the O2- and CO2-derived parameters the DeltaPCO2/C(a-v)O2 ratio had the highest correlation with the arterial lactate level ( r=0.57) . Moreover, for a threshold value of 1.4 the DeltaPCO2/C(a-v)O2 ratio predicted significantly better than the other parameters (receiver operating characteristic curves) the presence of hyperlactatemia (positive and negative predictive values of 86% and 80%, respectively) . The overall survival estimate at 1 month was greater when the DeltaPCO2/C(a-v)O2 ratio was less than 1.4 on the first set of measurements (38+/-10% vs . 20+/-8%, p<0.01) . CONCLUSION: The DeltaPCO2/C(a-v)O2 ratio seems a reliable marker of global anaerobic metabolism . Its calculation would be helpful for a better interpretation of pulmonary artery catheter data. Proc Natl Acad Sci U S A, 2002 Mar 19, 99(6), 3914 - 9 The quantity of nitric oxide released by macrophages regulates Chlamydia-induced disease; Huang J et al.; Intracellular bacteria of the genus Chlamydia cause numerous typically chronic diseases, frequently with debilitating sequelae . Genetic determinants of disease susceptibility after infection with Chlamydia bacteria are unknown . C57BL/6 mice develop severe pneumonia and poor immunity against Chlamydia after moderate respiratory infection whereas BALB/c mice are protected from disease and develop vigorous Th1 immunity . Here we show that infected C57BL/6 macrophages release more NO synthesized by NO synthase 2 (NOS2) than BALB/c macrophages and have lower mRNA concentrations of arginase II, a competitor of NOS2 for the common substrate, l-arginine . Reduction, but not elimination, of NO production by incomplete inhibition of NOS2 abolishes susceptibility of C57BL/6 mice to Chlamydia-induced disease . Thus, the quantity of NO released by infected macrophages is the effector mechanism that regulates between pathogenic and protective responses to chlamydial infection, and genes controlling NO production determine susceptibility to chlamydial disease. J Biol Chem, 2002 May 24, 277(21), 18658 - 64 Epub 2002 Mar 19. Genes essential to sodium-dependent bicarbonate transport in cyanobacteria: function and phylogenetic analysis; Shibata M et al.; The cyanobacterium Synechocystis sp . strain PCC 6803 possesses two CO(2) uptake systems and two HCO(3)(-) transporters . We transformed a mutant impaired in CO(2) uptake and in cmpA-D encoding a HCO(3)(-)transporter with a transposon inactivation library, and we recovered mutants unable to take up HCO(3)(-) and grow in low CO(2) at pH 9.0 . They are all tagged within slr1512 (designated sbtA) . We show that SbtA-mediated transport is induced by low CO(2), requires Na(+), and plays the major role in HCO(3)(-) uptake in Synechocystis . Inactivation of slr1509 (homologous to ntpJ encoding a Na(+)/K(+)-translocating protein) abolished the ability of cells to grow at {Na(+)} higher than 100 mm and severely depressed the activity of the SbtA-mediated HCO(3)(-) transport . We propose that the SbtA-mediated HCO(3)(-) transport is driven by DeltamuNa(+) across the plasma membrane, which is disrupted by inactivating ntpJ . Phylogenetic analyses indicated that two types of sbtA exist in various cyanobacterial strains, all of which possess ntpJ . The sbtA gene is the first one identified as essential to Na(+)-dependent HCO(3)(-) transport in photosynthetic organisms and may play a crucial role in carbon acquisition when CO(2) supply is limited, or in Prochlorococcus strains that do not possess CO(2) uptake systems or Cmp-dependent HCO(3)(-) transport. FEBS Lett, 2002 Feb 27, 513(2-3), 208 - 12 Induction of inducible nitric oxide synthase expression by 18beta-glycyrrhetinic acid in macrophages; Jeong HG et al.; Glycyrrhizin (GL), a triterpenoid saponin fraction of licorice, is reported to have anti-viral and anti-tumor activities and is metabolized to 18beta-glycyrrhetinic acid (GA) in the intestine by intestinal bacteria . However, the mechanism underlying its effects is poorly understood . To further elucidate the mechanism of GA, the aglycone of GL, we investigated the effects of GA on the release of nitric oxide (NO) and at the level of inducible NO synthase (iNOS) gene expression in mouse macrophages . We found that GA elicited a dose-dependent increase in NO production and in the level of iNOS mRNA . Since iNOS transcription has been shown to be under the control of the transcription factor nuclear factor kappaB (NF-kappaB), the effects of GA on NF-kappaB activation were examined . Transient expression assays with NF-kappaB binding sites linked to the luciferase gene revealed that the increased level of iNOS mRNA, induced by GA, was mediated by the NF-kappaB transcription factor complex . By using DNA fragments containing the NF-kappaB binding sequence, GA was shown to activate the protein/DNA binding of NF-kappaB to its cognate site, as measured by electrophoretic mobility shift assay . These results demonstrate that GA stimulates NO production and is able to up-regulate iNOS expression through NF-kappaB transactivation in macrophages. Water Res, 2002 Mar, 36(5), 1193 - 202 Factors affecting seasonal variation of membrane filtration resistance caused by Chlorella algae; Babel S et al.; A seasonal fluctuation pattern was observed in membrane filtration resistance by Chlorella algae cultured in open ponds in the tropical environment . In order to investigate the causes of this phenomenon, Chlorella was cultivated under controlled conditions and the cake resistance was measured by batch filtration in dead-end mode . The filtration resistance was found to be a function of environmental conditions . Algae could grow favourably and offered low specific cake resistance (R,s) on the order of 10(11) m/g for the culture temperature from 28 degrees C to 35 degrees C . The algal growth was inhibited and the specific cake resistance increased to the order of 10(12) m/g below or above this optimum temperature range . Strong solar radiation, coupled with high temperatures, also inhibited the growth of algae and resulted in higher specific cake resistance . The specific cake resistance of algae cultured at different temperatures increased with the amount of the extracellular organic matter (EOM) extracted by 0.1 N NaOH . Hence EOM, rather than bacteria present in the mono-algal culture, was considered to be the primary factor affecting the cake resistance . The specific cake resistance increased drastically after actively growing cells were stored in nutrient-free water under dark conditions . However, the resistance was slightly decreased when the algal cells were stored in NSIII nutrient media in a dark room, indicating the effect of nutrient availability on the change of the specific cake resistance under the light-limiting conditions . EOM extracted from the cells kept in the nutrient-free water contained less sugar than the fresh culture, whereas the EOM extracted from the cells stored in the NSIII media contained more sugar . The molecular distribution of the EOM shifted from below 1,000 kDa before storage to more than 2,000 kDa after storage in both the nutrient-free and NSIII media. Water Res, 2002 Mar, 36(5), 1127 - 34 Complete physico-chemical treatment for coke plant effluents; Ghose MK; Naturally found coal is converted to coke which is suitable for metallurgical industries . Large quantities of liquid effluents produced contain a large amount of suspended solids, high COD, BOD, phenols, ammonia and other toxic substances which are causing serious pollution problem in the receiving water to which they are discharged . There are a large number of coke plants in the vicinity of Jharia Coal Field (JCF) . Characteristics of the effluents have been evaluated . The present effluent treatment systems were found to be inadequate . Physico-chemical treatment has been considered as a suitable option for the treatment of coke plant effluents . Ammonia removal by synthetic zeolite, activated carbon for the removal of bacteria, viruses, refractory organics, etc . were utilized and the results are discussed . A scheme has been proposed for the complete physico-chemical treatment, which can be suitably adopted for the recycling, reuse and safe disposal of the treated effluent . Various unit process and unit operations involved in the treatment system have been discussed . The process may be useful on industrial scale at various sites. Acta Odontol Scand, 2002 Jan, 60(1), 10 - 2 Comparative analysis of some mouthrinses on the production of volatile sulfur-containing compounds; Rosing CK et al.; Oral malodor is mainly caused by the presence of volatile sulfur-containing compounds (VSC) produced by proteolytic periodontopathic bacteria in the oral cavity . Different solutions have been used as mouthrinses, trying to reduce malodor, and a large number is on the market . The aim of this study was to compare the effect of three commercially available mouthrinses with a simple inexpensive solution of zinc (zinc acetate 0.1%) on the production of VSC in vivo . Two of the solutions contained triclosan