Med Microbiol Immunol (Berl), 2002 May, 191(1), 5 - 10 Asialo-GM1 and asialo-GM2 are putative adhesion molecules for Moraxella catarrhalis; Ahmed K et al.; Moraxella catarrhalis is an important pathogen of respiratory and middle ear infections . We previously reported that the attachment of M . catarrhalis to pharyngeal epithelial cells is mediated by ganglioside M2 (GM2) . Several sets of adhesins or receptors are involved in such attachment process . In this study, we used the same strains and similar bacterial culture conditions as those in our previous study, and demonstrated by thin layer chromatography that M . catarrhalis can also bind to asialo-GM1 (Gg4Cer) and asialo-GM2 (Gg3Cer) . GalNAcbeta1-->4Galbeta1 is a common sequence in both Gg4Cer and Gg3Cer, and in many respiratory bacteria, this sequence acts as a receptor for attachment to host cells . Treatment of human pharyngeal epithelial cells with anti-GM2 and anti-Gg4Cer antibodies significantly decreased attachment of M . catarrhalis to these cells; however, treatment with anti-Gg3Cer antibody did not decrease M . catarrhalis attachment . Immunofluorescence microscopy revealed that human pharyngeal epithelial cells are positive for GM2 and Gg4Cer, but not for Gg3Cer . Our results indicate that Gg4Cer on human pharyngeal epithelial cells, and Gg3Cer,possibly on other cells, could serve as molecules for attachment of M . catarrhalis. J Forensic Sci, 2002 Jul, 47(4), 757 - 72 Characterization and validation studies of powerPlex 2.1, a nine-locus short tandem repeat (STR) multiplex system and penta D monoplex; Levedakou EN et al.; In order to increase the power of discrimination for human identification purposes, a nine-locus short tandem repeat (STR) multiplex, the GenePrint PowerPlex 2.1 system (PowerPlex 2.1) developed by Promega Corporation and a separate pentanucleotide-repeat locus, Penta D, were tested . This megaplex system includes the highly polymorphic loci FGA, TPOX, D8S1179, vWA, Penta E, D18S51, D21S11, TH01, and D3S1358 and may be used in combination with the eight-locus STR multiplex, the GenePrint PowerPlex 1.1 system (PowerPlex 1.1) that has been previously developed . Three of the loci, TPOX, TH01 and vWA, have been included in both systems for quality control purposes . As with PowerPlex 1.1, PowerPlex 2.1 is also based on a two-color detection of fluorescent-labeled DNA products amplified by polymerase chain reaction (PCR) and provides a valuable tool for accurate and rapid allele determination . The primer sequences used in the PowerPlex 2.1/Penta D system are also presented in this report . To meet the "Quality Assurance Standards for Forensic DNA Testing Laboratories" (FBI), we tested the efficiency and reproducibility of the PowerPlex 2.1/PentaD system by several validation studies that were conducted as a joint project among seven laboratories . Validation tests included concordance studies, sensitivity, and species specificity determination, as well as performance in forensic and environmentally impacted samples . The results produced from these tests demonstrated the consistency and reliability of the PowerPlex 2.1/Penta D system. Kansenshogaku Zasshi, 2002 Jun, 76(6), 425 - 31 {Pathogenicities of Mycobacterium intracellulare and M . avium strains to the mice which were isolated from non-tuberculous mycobactriosis patients}; Goto Y et al.; The virulence of 5 strains of M . intracellulare and 6 strains of M . avium to mice were examined . The former bacteria were obtained from the patients with mycobacterial pulmonary disease and the latter were from AIDS patients respectively . C57BL/6 (NRAMP-1 susceptible) and its NRAMP-1 congenic mice (resistant) were used to evaluate the virulence of these bacteria . Three of the 5 strains of M . intracelllulare showed a relatively high virulence . They grew in the liver, spleen and lungs of susceptible mice and even in the lungs of resistant mice . On the other hand, none of 6 strains of M . avium could grow in either susceptible or resistant mice . In conclusion, our mouse model may be a useful tool for evaluating the virulence of bacteria isolated from mycobacteriosis patients. Nucleic Acids Res, 2002 Jul 15, 30(14), 3067 - 77 A genetic screen identifies novel non-compatible loxP sites; Langer SJ et al.; The ability of the Cre/lox system to make precise genomic modifications is a tremendous accomplishment . However, recombination between cis-linked heterospecific lox sites limits the use of Cre- mediated exchange of DNA to systems where genetic selection can be applied . To circumvent this problem we carried out a genetic screen designed to identify novel mutant spacer-containing lox sites displaying enhanced incompatibility with the canonical loxP site . One of the mutant sites recovered appears to be completely stable in HEK293 cells constitutively expressing Cre recombinase and supports recombinase-mediated cassette exchange (RMCE) in bacteria and mammalian cell culture . By preventing undesirable recombination, these novel lox sites could improve the efficiency of in vivo gene transfer. FEBS Lett, 2002 Jul 31, 524(1-3), 127 - 33 Transient interaction of cpSRP54 with elongating nascent chains of the chloroplast-encoded D1 protein; 'cpSRP54 caught in the act'; Nilsson R et al.; The signal recognition particle (SRP) in bacteria and endoplasmic reticulum is involved in co-translational targeting . Plastids contain cpSRP54 and cpSRP43, unique to plants, but lack a SRP RNA molecule . A role for cpSRP in biogenesis of plastid-encoded membrane proteins has not been firmly established yet . In this study, a transient interaction between cpSRP54 and elongating D1 protein was observed using a homologous chloroplast translation system . Using the novel approach of cross-linking at different time points during elongation of full-length D1 protein, we showed that cpSRP54 interacts strongly with the elongating nascent chain forming two distinct cross-linked products . However, this interaction did not lead to an elongation arrest and cpSRP54 was released from the nascent chains, once they were longer than approximately 14 kDa . Detailed mutant analysis showed that the cpSRP54 interaction occurred via the first transmembrane domain, which could be replaced by other hydrophobic domains of more than 10 amino acids. Am J Trop Med Hyg, 2002 Jan, 66(1), 108 - 11 Field prevalence of Wolbachia in the mosquito vector Aedes albopictus; Kitrayapong P et al.; The endosymbiotic bacteria in the genus Wolbachia have been proposed as a potential candidate to deliver pathogen-blocking genes into natural populations of medically important insects . The successful application of Wolbachia in insect vector control depends on the ability of the agent to successfully invade and maintain itself at high frequency under field conditions . Here, we evaluated the prevalence of Wolbachia infections in a field population of the Wolbachia-superinfected mosquito Aedes albopictus . A field prevalence of 100% (n = 1,016) was found in a single population in eastern Thailand via polymerase chain reaction (PCR) testing of Wolbachia both from individual parent females and their corresponding F1 offspring . This is the first report of accurate Wolbachia prevalence in a field population of an insect disease vector . The prevalence of superinfection was estimated to be 99.41% . All single-infected individual mosquitoes (n = 6) were found to harbor group A Wolbachia . For this particular population, none was found to be single-infected with group B Wolbachia . Our results also show that PCR testing of field materials alone without checking F1 offspring overestimated the natural prevalence of single infection . Thus, the confirmation of infection status by means of F1 offspring was critical to the accurate estimates of Wolbachia prevalence under field conditions. Am J Trop Med Hyg, 2002 Jan, 66(1), 103 - 7 Maternal transmission efficiency of Wolbachia superinfections in Aedes albopictus populations in Thailand; Kittayapong P et al.; We examined the transmission efficiency of 2 strains of Wolbachia bacteria that cause cytoplasmic incompatibility in field populations of Aedes albopictus by polymerase chain reaction assay . We found mainland and island populations throughout Thailand to be superinfected with group A and B bacteria . Of 320 Wolbachia-positive adult mosquitoes, 97.5% were infected with both groups . Single infected individuals of each Wolbachia group were encountered in nearly equal numbers . We screened 550 offspring from 80 field-collected mothers and found the transmission efficiency of group A Wolbachia to be 96.7% and that of group B Wolbachia to be 99.6% . Mothers that did not transmit both Wolbachia infections to all of their offspring were significantly larger in size than those with perfect transmission fidelity . We discuss our findings in relation to the prospects of the use of Wolbachia as a gene-driving mechanism. Arch Pharm Res, 2002 Jun, 25(3), 258 - 69 Synthesis of new 2-thiouracil-5-sulfonamide derivatives with biological activity; Fathalla A et al.; 2-Thiouracil-5-sulfonylchloride 1 reacted with a series of aromatic and heterocyclic amines to give 2a-j . The same compound 1 was reacted with a series of sulphonamides giving different sulphonamides of type 3a-e . On the other hand compound 1 was allowed to react with p-aminoacetophenone givining compound 4 which in turn was allowed to react with derivatives of alkyl thiosemicarbazides to give thiosemicarbazones of type 5a-e, also compound 4 was monobrominated to give compound 6 which in turn was reacted thiosemicarbazones of some aldehydes to give the corresponding thiazole derivatives 7a-f . In the same time compound 4 was reacted with a series of aromatic and heterocyclic aldehydes givining chalcones 8a-g (Claisen-Schemidt reaction) . Also compound 4 was allowed to react with a series of aromatic and heterocyclic aldehydes, ethyl cyano acetate and/or malononitrile, and ammonium acetate giving pyridine derivatives 9a-d and 10a-e respectively . The biological effects of some of the new synthesized compounds was also investigated. Pharm Res, 2002 Jun, 19(6), 825 - 31 Transport and utilization of arginine and arginine-containing peptides by rat alveolar macrophages; Yang XD et al.; PURPOSE: To demonstrate that rat alveolar macrophages (AM) exhibited the PepT1-like transporter for the uptake of arginine (Arg)-containing small peptides and utilized these peptides as direct substrates for nitric oxide (NO) production . NO is an important mediator that, on one hand, protects the lung from bacteria infection and, on the other hand, augments inflammatory lung injury . METHOD: The uptake of small peptides by rat AM was evaluated using fluorescein isothiocyanate (FITC)-labeled (*) peptides (Arg-Lys*, Gly-Sar-Lys*, and beta-Ala-Lys*), high-performance liquid chromatography (HPLC) analysis of potential peptide degradation, and known inhibitors of Arg and PepT1 transport . NO production by AM through Arg and Arg-containing peptides was studied with and without inhibition by transport inhibitors . The presence of PepT1-like transporter on AM was evaluated using anti-PepT1 antisera and Western blot analysis . The substrate specificity of Arg-Gly and Arg-Gly-Asp was determined using purified inducible NO synthase (iNOS) . The availability of Arg-containing peptides in the lung was determined by HPLC analysis of bronchoalveolar lavage (BAL) fluid . RESULTS: The FITC-labeled peptides were internalized by AM without degradation . The uptake of Arg-Lys*, beta-Ala-Lys*, and Gly-Sar-Lys* was blocked (approximately 50%) by cephradine (an inhibitor of PepT1 for peptide transport) but not by Lys (an inhibitor on cationic amino acid transporter 2B for Arg transport) . The NO production by AM through Arg-containing peptides was significantly blocked only by PepT1 inhibitors and by an anti-PepT1 antibody in a dose-dependent manner . These inhibitors had no effect on the AM production of NO using Arg as a substrate . Arg-Gly and Arg-Gly-Asp were found to be direct substrates for iNOS with similar Km and Vmax values to those of Arg . But, the production of NO by AM using these peptides as substrates was 2-fold higher than using Arg as a substrate . Both Arg-Gly and Arg-Gly-Asp were found in the BAL fluid . The presence of a PepT1-like transporter on AM was confirmed by Western blotting . CONCLUSION: This study shows that AM exhibit PepT1-like transporter for small peptide uptake . Arginine-containing peptides, through the PepT1 transporter system, can serve as direct substrates of iNOS for the production of NO by AM. J Immunol Methods, 2002 Aug 1, 266(1-2), 197 - 207 Isolation and characterization of rabbit single chain antibodies to human Reg Ialpha protein; Chi XS et al.; To investigate the role of Reg Ialpha in human inflammatory bowel disease (IBD), we made two phage-displayed single chain variable fragment (scFv) libraries from rabbits immunized with recombinant or native human Reg Ialpha . After one to three rounds of panning, we were able to isolate phage-displaying scFvs, which bound to human Reg Ialpha . Anti-Reg Ialpha scFvs from both libraries showed similar immunoreactivity to different processed forms of the protein . Despite several DNA fingerprint patterns among these clones, their deduced amino acid sequences are highly homologous with 100% identity in the complementarity-determining regions (CDRs) of the variable segment of heavy chain (VH) region and a small variation in the CDR1 of the variable segment of light chain (VL) region . We also expressed and purified soluble myc-tagged or glutathione S-transferase (GST) fusion scFv proteins from bacteria . Immunohistochemical studies using one of our anti-Reg Ialpha scFv antibodies showed prominent staining in the metaplastic Paneth cell population and light staining in the lamina propria . This scFv antibody is now being used for studies of the role of Reg Ialpha in human IBD. Z Naturforsch {C}, 2002 May-Jun, 57(5-6), 516 - 21 Discriminatory power of RAPD, PCR-RFLP and southern blot analyses of ureCD or ureA gene probes on Helicobacter pylori isolates; Smith S et al.; The genetic diversity of 33 Nigerian Helicobacter pylori isolates were studied using RAPD, PCR-RFLP and Southern blot analysis of ureA or ureCD gene probes . RAPD was able to distinguish the following number of isolates using the primers 3880: 5'-AAGAGCCCGT-3' (28), 3881 :5'-AACGCGCAAC-3' (33) and OPH8 :5'-GAAACACCCC-3' (25) . Southern blot analysis using the ureCD probe was also able to distinguish the 12 isolates tested into ten different patterns . The PCR-RFLP technique distinguished all 33 isolates into six types . In conclusion, considering typeability, discriminatory power, and convenience, RAPD with the 3881 primer was considered the most useful technique. Biol Pharm Bull, 2002 Jul, 25(7), 861 - 6 Prevention of growth and metastasis of murine melanoma through enhanced natural-killer cytotoxicity by fatty acid-conjugate of protopanaxatriol; Hasegawa H et al.; Ginsenosides, the glycosides of Panax ginseng, are metabolized (deglycosylated) by intestinal bacteria after oral administration . 20(S)-Protopanaxatriol (M4) is the main bacterial metabolite of protopanaxatriol-type ginsenosides and mediates their antitumor effects . To clarify the mechanism of the M4-mediated antitumor effect, the antitumor activity and metabolism of M4 was examined, using the C57BL/6 mice implanted with B16-BL6 melanoma . The chronic oral administration of M4 inhibited the growth of B16-BL6 melanoma at the implanted site . Analyses using TLC, HPLC, MS and NMR suggest that orally administered M4 was absorbed from the small intestine into the mesenteric lymphatics followed by the rapid esterification of M4 with fatty acids and its accumulation in the tissues including the liver and lung . The administration of M4 prior to the intravenous injection of B16-BL6 cells abrogated the enhanced lung metastasis in the mice pretreated with 2-chloroadenosine more effectively than in those pretreated with anti-asialo GM1 . The esterified M4 (EM4) did not directly affect tumor growth in vitro, whereas it stimulated splenic NK cells to become cytotoxic to tumor cells . These results indicate that the antitumor activity of M4 is based on the NK cell-mediated tumor lysis enhanced by EM4. Salud Publica Mex, 2002 May-Jun, 44(3), 201 - 6 {Acute respiratory infections in children attending a child day care center}; Nandi-Lozano E et al.; OBJECTIVE: To assess the incidence of acute respiratory infections and bacterial colonization in children attending a daycare center . MATERIAL AND METHODS: A cohort study was conducted from April to Octuber 1999, among 85 children aged under four years, who attended the daycare center at Hospital Infantil de Mexico (Mexico City's Children's Hospital) "Federico Gomez" . Acute respiratory infection incidence rates and quarterly point prevalence figures of nasopharyngeal colonization were obtained . Data were analyzed using descriptive statistics . RESULTS: A total of 85 children were studied (40 girls and 45 boys) during 9,090 children-days of follow-up . Three children had a history of atopia (3.5%), six a history of asthma (7%), and 39 (46%) were exposed to passive smoking . There were 258 events of respiratory tract infection for an incidence rate of 10.3 infections per person-year (95% CI 8.7-12.0) . The main clinical syndromes were pharyngitis (95%), acute otitis media (3.5%), and bronchiolitis (1%) . The incidence rates of otitis and bronchiolitis were 0.36 and 0.12 per child-year of observation, respectively . The prevalence figures of nasopharyngeal colonization for the three main bacteria were: S . pneumoniae 20.4%; nontypable H . influenzae 13%; and Moraxella catarrhalis 8% . CONCLUSIONS: Study results show a high prevalence of colonization due to invasive strains, as well as a two-fold incidence rate of acute respiratory infection, higher than those reported in community surveys . These results add to the description of this poorly documented infectious disease in Mexico . The English version of this paper is available at: http://www.insp.mx/salud/index.html. J Prosthet Dent, 2002 Jun, 87(6), 674 - 8 Endodontic failure caused by inadequate restorative procedures: review and treatment recommendations; Heling I et al.; A review of the literature was performed to determine whether prompt placement of coronal restorations, including sealing and placement of posts and cores, can positively influence the long-term prognosis of teeth after root canal therapy . Both hand and MEDLINE searches were employed to identify peer-reviewed articles on radicular apical integrity after coronal restorations, especially where root canal space was used for post and core fabrication . A total of 41 articles published between 1969 and 1999 (the majority from the 1990s) were reviewed . The literature suggests that the prognosis of root canal-treated teeth can be improved by sealing the canal and minimizing the leakage of oral fluids and bacteria into the periradicular areas as soon as possible after the completion of root canal therapy. Chem Pharm Bull (Tokyo), 2002 Jul, 50(7), 892 - 5 An in vitro and in vivo investigation into the suitability of bacterially triggered delivery system for colon targeting; Raghavan CV et al.; The colon specific drug delivery systems based on polysaccharides; locust bean gum and chitosan in the ratio of 2 : 3, 3 : 2 and 4 : 1 were evaluated using in vitro and in vivo methods . The in vitro studies in pH 6.8 phosphate buffer containing 2% w/v rat caecal contents showed that the cumulative percentage release of mesalazine after 26 h were 31.25+/-0.56, 46.25+/-0.96, 97.5+/-0.26 (mean+/-S.D.), respectively . The in vivo studies conducted in nine healthy male human volunteers for the various formulations revealed that, the drug release was initiated only after 5 h (i.e.) transit time of small intestine and the bioavailability (AUC(0-->t*)) of the drug was found to be 85.24+/-0.10, 196.08+/-0.12, 498.62+/-0.10 microg x h/ml 26 (mean+/-S.D.), respectively . These studies on the polysaccharides demonstrated that the combination of locust bean gum and chitosan as a coating material proved capable of protecting the core tablet containing mesalazine during the condition mimicking mouth to colon transit . In particular, the formulation containing locust bean gum and chitosan in the ratio of 4 : 1 held a better dissolution profile, higher bioavailability and hence a potential carrier for drug targeting to colon. Zoolog Sci, 2002 Jun, 19(6), 695 - 701 A comparative study of body wall structures of a pogonophore and an annelid from a phylogenetic viewpoint; Matsuno A et al.; Pogonophores are tube worms that live in reducing deep-sea waters where sunlight does not penetrate . They are highly adapted for their special habitat in lacking guts and possessing endosymbiotic chemosynthetic bacteria . Because of these peculiar characteristics, it is not yet clear whether they should be classified as annelids or not . Electron-microscopic observations of sections of a Japanese pogonophore (Oligobrachia mashikoi) show that the body wall has circular and longitudinal muscular systems . These muscular systems, however, differ from the annelid (Branchiura sowerbyi) in these ways: (1) The outer circular muscle of the pogonophore was constructed of smooth muscle cells . In contrast, that of the annelid was composed of obliquely-striated muscle cells, even though the cells were small and bore undeveloped characteristics . (2) The inner longitudinal muscle of the pogonophore was constructed of undeveloped obliquely-striated muscle cells, whereas that of the annelid was composed of well-developed ones . These observations suggest that this pogonophore can not be classified as an annelid, although many previous studies have placed pogonophores in that phylum. J Biol Chem, 2002 Sep 27, 277(39), 36351 - 6 Epub 2002 Jul 18. NMR structure of PW2 bound to SDS micelles . A tryptophan-rich anticoccidial peptide selected from phage display libraries; Tinoco LW et al.; PW2 (HPLKQYWWRPSI) was selected from phage display libraries through an alternative panning method using living sporozoites of Eimeria acervulina as target . Synthetic PW2 shows anticoccidial activity against E . acervulina and Eimeria tenella with very low hemolytic activity . It also displays antifungal activity but no activity against bacteria . We present the solution structure of the PW2 bound to SDS micelles . In the absence of an interface, PW2 is in random coil conformation . In micelles, structural calculation shows that Trp-7 forms the hydrophobic core that is important for the peptide folding . Lys-4, Tyr-6, Trp-8, and Arg-9 are in the same surface, possibly facing the micelle interface . This possibility was supported by the fact that chemical shift differences for these residues were more pronounced when compared with PW2 in water and in SDS . PW2 gains structure upon binding to SDS micelles . Lys-4, Tyr-6, Trp-8, and Arg-9 were found to bind to the micelle . Trp-7, Trp-8, and Arg-9 composed the WW+ consensus found in the sequence of the peptides selected with the phage display technique against E . acervulina sporozoites . This suggested that Trp-7, Trp-8, and Arg-9 are probably key residues not only for the peptide interaction with SDS micelles but also for the interaction with E . acervulina sporozoites surface. Comp Biochem Physiol B Biochem Mol Biol, 2002 Aug, 132(4), 739 - 50 Proline biosynthesis genes and their regulation under salinity stress in the euryhaline copepod Tigriopus californicus; Willett CS et al.; Diverse organisms regulate concentrations of intracellular organic osmolytes in response to changes in environmental salinity or desiccation . In marine crustaceans, accumulation of high concentrations of proline is a dominant component of response to hyperosmotic stress . In the euryhaline copepod Tigriopus californicus, synthesis of proline from its metabolic precursor glutamate is tightly regulated by changes in environmental salinity . Here, for the first time in a marine invertebrate, the genes responsible for this pathway have been cloned and characterized . The two proteins display the sequence features of homologous enzymes identified from other eukaryotes . One of the cloned genes, delta1-pyrroline-5-carboxylase reductase (P5CR), is demonstrated to have the reductase enzyme activity when expressed in proline-auxotroph bacteria, while the second, delta1-pyrroline-5-carboxylase synthase (P5CS), does not rescue proline-auxotroph bacteria . In contrast to results from higher plants, neither levels of P5CS nor P5CR mRNAs increase in response to salinity stress in T . californicus . Hence, regulation of proline synthesis during osmotic stress in T . californicus is likely mediated by some form of post-transcriptional regulation of either P5CS or P5CR . Understanding the regulation this pathway may elucidate the mechanisms limiting the salinity ranges of marine taxa . Microb Pathog, 2002 Jul, 33(1), 7 - 15 Previous infection with the nematode Nippostrongylus brasiliensis alters the immune specific response against Chlamydophila abortus infection; Buendia A et al.; An experimental mouse model to analyze the interaction between the immune responses elicited following infection with Nippostrongylus brasiliensis and Chlamydophila abortus has been established . Mice infected with C . abortus 7 days after N . brasiliensis showed an increased bacterial multiplication in spleen and liver compared to bacteria-alone infected mice . However the morbidity of these mice, expressed as weight loss, was significantly lower . Analysis of the immune responses elicited showed that spleen from co-infected mice had reduced IFN-gamma production in response to C . abortus antigen . The bias towards a type 2 response in co-infected mice was confirmed by an increase in the production of IL-4 and in the lower ratio IgG2a/IgG1 . In pregnant mice co-infection caused a delay in the time of abortion and an increased systemic susceptibility to C . abortus infection. Vaccine, 2002 Jul 26, 20(23-24), 2981 - 8 Serum-derived IgG1-mediated immune exclusion as a mechanism of protection against H . pylori infection; Keenan J et al.; The induction of protective immunity against Helicobacter challenge in a murine model was found to correlate with the magnitude of IgG (serum and gastric lavage) responsiveness to intra-nasal (i.n.) immunisation . IgG1-secreting hybridoma backpacks in Helicobacter pylori (H . pylori)-infected mice revealed serum transudation into the stomach . A Lpp20-specific monoclonal antibody was associated with significantly reduced H . pylori colonisation . Histology revealed aggregates of the remaining H . pylori in these mice, suggesting a role for IgG1-mediated immune exclusion of the bacteria . In vitro immunogold electron microscopy supported this hypothesis, but also suggested that a threshold of H . pylori-specific antibody needs to be maintained if immune exclusion by the host is to overcome immune evasion by the bacteria. Toxicology, 2002 Aug 1, 177(1), 11 - 22 Tyrosyl radical production by myeloperoxidase: a phagocyte pathway for lipid peroxidation and dityrosine cross-linking of proteins; Heinecke JW; To kill invading bacteria, viruses, and fungi, phagocytes secrete hydrogen peroxide (H(2)O(2)) and the heme enzyme myeloperoxidase . We have explored the possibility that myeloperoxidase might use H(2)O(2) to convert L-tyrosine to tyrosyl radical . Activated human neutrophils and monocytes used the system to oxidize free L-tyrosine to o,o'-dityrosine, a stable product of tyrosyl radical . Protein-bound tyrosyl residues exposed to myeloperoxidase, H(2)O(2), and L-tyrosine were also oxidized to o,o'-dityrosine . The cross-linking reaction required free L-tyrosine, suggesting that myeloperoxidase converts the amino acid to a diffusible radical catalyst that promotes protein oxidation . We used electron paramagnetic resonance to provide direct evidence that the oxidizing intermediate is free tyrosyl radical . Myeloperoxidase-generated tyrosyl radical also initiates lipid peroxidation, suggesting that activated phagocytes might also be able to oxidize lipids in host tissues . Moreover, myeloperoxidase is present and active in human atherosclerotic tissue, and levels of protein-bound dityrosine are elevated in such lesions . Our recent studies indicate that activated neutrophils use oxidants generated by the phagocyte NADPH oxidase to produce protein-bound dityrosine during acute inflammation . Collectively, these findings suggest that generation of tyrosyl radical by myeloperoxidase allows activated phagocytes to damage both proteins and lipids . Elevated levels of o,o'-dityrosine have been detected in inflammatory lung disease, neurodegenerative disorders, and aging . Thus, oxidation of tyrosine to tyrosyl radical might play a role in the pathogenesis of many diseases. Free Radic Biol Med, 2002 Aug 1, 33(3), 323 - 36 Molecular and cellular mechanisms involved in Helicobacter pylori-induced inflammation and oxidative stress; Naito Y et al.; Helicobacter pylori (H . pylori)-infection leads to different clinical and pathological outcomes in humans, including chronic gastritis, peptic ulcer disease, and gastric neoplasia . The key determinants of these outcomes are the severity and distribution of the H . pylori-induced inflammation . Antral-type gastritis is associated with excessive acid secretion and a high risk of duodenal ulcer . In contrast, gastritis that involves the acid-secreting corpus region leads to hypochlorhydria, progressive gastric atrophy, and an increased risk of gastric cancer . The key pathophysiological event in H . pylori infection is initiation and continuance of an inflammatory response . Bacteria or their products trigger this inflammatory process and the main mediators are cytokines . Identification of both host- and bacterial-factors that mediate is an intense area of interest in current researches . Recent data indicates that the cag pathogenicity island plays a crucial role in H . pylori-induced gastric inflammation via the activation of gene transcription . It has been demonstrated that oxidative and nitrosative stress associated with inflammation plays an important role in gastric carcinogenesis as a mediator of carcinogenic compound formation, DNA damage, and cell proliferation . Genetic information regulating such stress would be one of the host factors determining the outcome--particularly when the outcome is gastric cancer--of H . pylori infection, and the compound that attenuates such stress may be a candidate for use in chemoprevention . This review highlights recent advances in understanding of the mechanisms underlying chronic inflammation following infection with H . pylori. J Org Chem, 2002 Jul 26, 67(15), 5416 - 8 Synthesis of 4-diphosphocytidyl-2-C-methyl-D-erythritol and 2-C-methyl-D-erythritol-4-phosphate; Koppisch AT et al.; 2-C-Methyl-D-erythritol 4-phosphate (MEP, 2) and 4-diphosphocytidyl-2-C-methyl-D-erythritol (CDPME, 3) are metabolites in the MEP pathway for biosynthesis of isoprenoid compounds in bacteria, plant chloroplasts, and algae . The free phosphoacid of 2 was prepared from benzyloxyacetone in five steps with an overall yield of 27% and an enantiomeric ratio (er) of 75:25 . Following titration to the corresponding tributylammonium salt, 2 was coupled to cytidine 5'-monophosphate using a protocol originally developed for synthesis of base-sensitive nucleoside diphosphate sugars to give 3 in 40% yield, following purification by size exclusion chromatography. J Mol Microbiol Biotechnol, 2002 Jul, 4(4), 405 - 15 Bioinformatic analyses of the tRNA: (guanine 26, N2,N2)-dimethyltransferase (Trm1) family; Bujnicki JM et al.; Functional analyses of the tRNA:(guanine 26, N2,N2)-dimethyltransferase (Trm1) have been hampered by a lack of structural information about the enzyme and by low sequence similarity to better studied methyltransferases . Here we used computational methods to detect novel homologs of Trm1, infer the evolutionary relationships of the family, and predict the structure of the Trm1 methyltransferase . The N-terminal region of the protein is predicted to form an S-adenosylmethionine-binding domain, which harbors the active site . The C-terminal region is rich in predicted alpha-helices and, in analogy to other nucleic acid methyltransferases, may constitute the target recognition domain of the enzyme . Interposing these two domains, most Trm1 homologs possess a highly variable inserted sequence that is delimited by a Cys4 cluster, likely forming a Zn-finger structure . The residues of Trm1 predicted to participate in cofactor binding, target recognition, and catalysis, were mapped onto a preliminary structural model, providing a platform for designing new experiments to better understand the molecular functions of this protein family . In addition, identification of novel, atypical Trm1 homologs suggests candidates for cloning and biochemical characterization. Ann Dermatol Venereol, 2002 May, 129(5 Pt 1), 728 - 31 {Mycobacterial bovis BCG cutaneous infections following mesotherapy: 2 cases}; Marco-Bonnet J et al.; INTRODUCTION: Infectious complications following mesotherapy are usually due to ordinary bacteria or atypical mycobacteria . We report two new cases of mycobacterial bovis BCG infections following mesotherapy . To our knowledge only one case has already been reported.CASES REPORTS: A 52 year-old woman developed vaccinal MERIEUX BCG cutaneous abscesses following mesotherapy . Identification was made by a novel class of repeated sequences: Mycobacterial interspersed repetitive units . Despite prolonged anti-tuberculous therapy, complete remission was not obtained and surgical excision was performed . The second case was a 49 year-old man who developed a mycobacterial bovis BCG cutaneous abscess (Connaught) after mesotherapy, the regression of which was obtained with anti-tuberculous therapy.DISCUSSION: The severity of these two mycobacterial infections following mesotherapy illustrate the potential risks of mesotherapy . Identification is possible by molecular biology techniques (PCR and sequencing) . The origin of this infection is unclear and therapeutic decision is difficult . Some authors recommend anti-tuberculous therapy but surgical excision may be necessary as in our cases. Anal Biochem, 2002 Jul 15, 306(2), 171 - 80 23Na NMR study of Fibrobacter succinogenes S85: comparison of three chemical shift reagents and calculation of sodium concentration using ionophores; Delort AM et al.; In order to measure intracellular sodium concentrations in resting cells of Fibrobacter succinogenes S85 by (23)Na NMR spectrometry, two methodological aspects were studied . First, three different shift reagents (Dy(PPP(i))(7-)(2), Tm(DOTP)(5-), and Dy(TTHA)(3-)) were tested for their ability to separate internal and external (23)Na NMR resonances . Their toxicity toward F . succinogenes cells was evaluated by in vivo(13)C NMR experiments . Tm(DOTP)(5-) was found to be the most efficient shift reagent while being nontoxic . Second, a new methodology was developed to calculate intracellular sodium concentration in F . succinogenes by using ionophores . This approach avoided the problem of intracellular volume measurement and that of sodium visibility determination. Mol Microbiol, 2002 Jul, 45(2), 365 - 74 Role of the extracytoplasmic-function sigma factor sigma(H) in Mycobacterium tuberculosis global gene expression; Manganelli R et al.; Like other bacterial species, Mycobacterium tuberculosis has multiple sigma (sigma) factors encoded in its genome . In previously published work, we and others have shown that mutations in some of these transcriptional activators render M . tuberculosis sensitive to various environmental stresses and, in some cases, cause attenuated virulence phenotypes . In this paper, we characterize a M . tuberculosis mutant lacking the ECF sigma factor sigma(H) . This mutant was more sensitive than the wild type to heat shock and to various oxidative stresses, but did not show decreased ability to grow inside macrophages . Using quantitative reverse transcription-PCR and microarray technology, we have started to define the sigma(H) regulon and its involvement in the global regulation of the response to heat shock and the thiol-specific oxidizing agent diamide . We identified 48 genes whose expression increased after exposure of M . tuberculosis to diamide; out of these, 39 were not induced in the sigH mutant, showing their direct or indirect dependence on sigma(H) . Some of these genes encode proteins whose predicted function is related to thiol metabolism, such as thioredoxin, thioredoxin reductase and enzymes involved in cysteine and molybdopterine biosynthesis . Other genes under sigma(H) control encode transcriptional regulators such as sigB, sigE, and sigH itself. Gastroenterol Clin North Am, 2002 Mar, 31(1), 63 - 76 The genetics of inflammatory bowel disease; Duerr RH; The complex genetics of IBD is characterized by more than one susceptibility locus, genetic heterogeneity, incomplete penetrance, and probable gene-gene and gene-environment interactions . Functional candidate gene association studies during the past few decades have revealed only modest associations between IBD and genetic variants in the HLA genes and a limited number of other genes that are involved in immune regulation and the inflammatory response . Important advances in IBD genetics research have come about from systematic genome searches for IBD loci . The identification of Crohn's disease-associated NOD2 genetic variants that appear to alter the innate immune response to bacteria is a seminal finding that perhaps is the greatest advance toward understanding the pathogenesis of IBD in decades . The future discovery of other IBD genetic risk factors, facilitated by the completion of the human genome sequencing and annotation, may allow the development of better therapies, possibly including preventive therapies, for patients with Crohn's disease and ulcerative colitis. Z Gastroenterol, 2002 Jul, 40(7), 525 - 9 Role of the intestinal epithelium in orchestrating innate and adaptive mucosal immunity; Maaser C et al.; The mucosa that lines the human colon and small intestine is a site of chronic regulated "physiologic" inflammation . This contrasts markedly with other mucosal sites in that if the numbers of T and B cells, eosinophils, mast cells, macrophages, and dendritic cells that are present in the human intestinal tract were to be present in other sites, those sites would be considered to be sites of chronic pathological inflammation . This review examines the role of the intestinal epithelium in the development of "physiologic" intestinal mucosal inflammation and focuses on its role in signalling and mediating host innate and adaptive mucosal immune responses. Proc Natl Acad Sci U S A, 2002 Aug 6, 99(16), 10417 - 22 Epub 2002 Jul 16. The threshold for polyglutamine-expansion protein aggregation and cellular toxicity is dynamic and influenced by aging in Caenorhabditis elegans; Morley JF et al.; Studies of the mutant gene in Huntington's disease, and for eight related neurodegenerative disorders, have identified polyglutamine (polyQ) expansions as a basis for cellular toxicity . This finding has led to a disease hypothesis that protein aggregation and cellular dysfunction can occur at a threshold of approximately 40 glutamine residues . Here, we test this hypothesis by expression of fluorescently tagged polyQ proteins (Q29, Q33, Q35, Q40, and Q44) in the body wall muscle cells of Caenorhabditis elegans and show that young adults exhibit a sharp boundary at 35-40 glutamines associated with the appearance of protein aggregates and loss of motility . Surprisingly, genetically identical animals expressing near-threshold polyQ repeats exhibited a high degree of variation in the appearance of protein aggregates and cellular toxicity that was dependent on repeat length and exacerbated during aging . The role of genetically determined aging pathways in the progression of age-dependent polyQ-mediated aggregation and cellular toxicity was tested by expressing Q82 in the background of age-1 mutant animals that exhibit an extended lifespan . We observed a dramatic delay of polyQ toxicity and appearance of protein aggregates . These data provide experimental support for the threshold hypothesis of polyQ-mediated toxicity in an experimental organism and emphasize the importance of the threshold as a point at which genetic modifiers and aging influence biochemical environment and protein homeostasis in the cell. Oper Dent, 2002 Jul-Aug, 27(4), 330 - 42 Histopathologic study on pulp response to single-bottle and self-etching adhesive systems; Medina VO 3rd et al.; This study compared the pulp response to seven adhesive resins (three single-bottle and four self-etching primers) and their companion resin composite systems with a commercial calcium hydroxide material when applied to exposed monkey pulps . The control group was capped with Dycal (DY), while the experimental groups were capped with one of the following adhesive resin systems: AQ Bond (AQ), Single Bond (SB), Imperva Fluorobond (IF), One Step (OS), Prime&Bond NT (PBNT), Perme Bond F (PBF) and One-up Bond F (OBF) . Histopathologic evaluation of pulp tissue disorganization, inflammatory cell infiltration, reparative dentin formation and bacterial penetration at the 3rd, 30th and 90th post-operative days was done using light microscopy . Data were analyzed using the Kruskal-Wallis test followed by the Least Significant Difference Test to determine differences between the control group and the experimental groups at each observation period . The correlation of inflammatory cell infiltration and bacterial presence was investigated by the Kendall correlation analysis . All tests were performed at a 95% level of confidence . The pulpal responses of groups DY, SB, OS, PBF and OBF were generally characterized by none-to-mild pulp tissue disorganization and inflammatory cell infiltration . Also, initiation of reparative dentin formation was found earlier in Group DY, resulting in more complete dentin bridges at the 30- and 90-day observation periods . Groups AQ, IF and PBNT had significantly more inflammatory cell infiltration and a lower incidence of reparative dentin formation than Group DY . A significant correlation was detected between inflammatory cell infiltration and the presence of bacteria . It is concluded that the pulp response to SB, OS, PBF and OBF is not significantly different from the calcium hydroxide preparation . However, calcium hydroxide capping resulted in a higher incidence and faster rate of reparative dentin formation. Chem Commun (Camb), 2002 Feb 21, (4), 318 - 9 Redox control of the P450cam catalytic cycle: effects of Y96F active site mutation and binding of a non-natural substrate; Reipa V et al.; Spectroelectrochemistry measurements are used to demonstrate that active site mutation and binding of an non-natural substrate to P450cam (CYP101) reduces the shift in the redox potential caused by substrate-binding, and thereby results in slower catalytic turnover rate relative to wild-type enzyme with the natural camphor substrate. Int J Lepr Other Mycobact Dis, 2002 Mar, 70(1), 25 - 33 Light and ultrastructural study of sciatic nerve lesions induced using intraneural injection of viable Mycobacterium leprae in normal and immunosuppressed Swiss white mice; Shetty VP et al.; Freshly harvested M . leprae were microinjected into the sciatic nerves of nonimmunosuppressed (non-TR) and immunosuppressed (TR) mice using the technique described by Wisniewski and Bloom . The lesions thus induced, on bypassing the blood-nerve barrier, were biopsied at regular intervals beginning 24 hr and followed up to one year . The fate of M . leprae and the ensuing inflammation and nerve damage were studied using light and electron microscopy . The lesions in both non-TR and TR mice at 24 hr showed an influx of polymorphonuclear leukocytes and an increase in mast cells . The influx and peaking of lymphocytes were delayed by two weeks and 6 weeks, respectively, in TR mice, but the density of lymphocytes at the peak intervals was comparable in both . The plasma cells denoting the humoral response were seen in both, but there was a delay of 3 weeks in non-TR mice . The lesions in non-TR mice showed differentiation of macrophages into epithelioid cells and the formation of giant cells depicting borderline tuberculoid leprosy (BT), Whereas in TR mice, the macrophages showed foamy cytoplasmic changes depicting borderline lepromatous leprosy (BL) . Other significant observations common to both non-TR and TR mice were: a) The lesions remained highly localized and showed signs of regression at the 6th and the 12th month intervals . b) The characteristic segmental demyelination and some attempt at remyelination were seen at the site . c) The influx of lymphocytes concorded well with demyelination . d) Bacteria were only seen in the macrophages and never in the Schwann cells or endothelial cells . e) Bacteria persisted in the macrophages, but appeared progressively degenerate at the 6th and 12th post-inoculation months, suggesting loss of viability . The study shows that there was a very effective containment of the infection and that the Schwann cells were resistant to M . leprae infection in the neural milieu . Nerve damage and Schwann cell bacillation do not go hand-in-hand. Proc Natl Acad Sci U S A, 2002 Jul 23, 99(15), 10173 - 8 Epub 2002 Jul 15. Chaotic mixing deep in the lung; Tsuda A et al.; Our current understanding of the transport and deposition of aerosols (viruses, bacteria, air pollutants, aerosolized drugs) deep in the lung has been grounded in dispersive theories based on untested assumptions about the nature of acinar airflow fields . Traditionally, these have been taken to be simple and kinematically reversible . In this article, we apply the recently discovered fluid mechanical phenomenon of irreversible low-Reynolds number flow to the lung . We demonstrate, through flow visualization studies in rhythmically ventilated rat lungs, that such a foundation is false, and that chaotic mixing may be key to aerosol transport . We found substantial alveolar flow irreversibility with stretched and folded fractal patterns, which lead to a sudden increase in mixing . These findings support our theory that chaotic alveolar flow--characterized by stagnation saddle points associated with alveolar vortices--governs gas kinematics in the lung periphery, and hence the transport, mixing, and ultimately the deposition of fine aerosols . This mechanism calls for a rethinking of the relationship of exposure and deposition of fine inhaled particles. Atherosclerosis, 2002 Sep, 164(1), 153 - 60 Positive Chlamydia pneumoniae serology is associated with elevated levels of tumor necrosis factor alpha in patients with coronary heart disease; Schumacher A et al.; Infectious agents are possible stimulators of inflammation in atherogenesis . The aim of this study was to investigate if Chlamydia pneumoniae and Helicobacter pylori were associated with elevated levels of tumor necrosis factor alpha (TNFalpha) and interleukin-6 in coronary heart disease (CHD) patients (n=193) and age- and sex-matched controls (n=193) as markers of increased inflammatory activity . C reactive protein (CRP) and fibrinogen were also included . Serologic status towards the two bacteria was measured and levels of the inflammatory markers were compared between seropositives and seronegatives, each study group being evaluated separately . In CHD patients Chlamydia lipopolysaccharide (LPS) IgA seropositivity predicted elevated TNFalpha levels (P=0.009), still statistically significant after adjustment for traditional cardiovascular risk factors (P=0.005) . Chlamydia LPS IgG seropositivity independently predicted fibrinogen levels in CHD patients (P=0.028), while no association between serology and inflammatory markers were observed among controls . H . pylori seropositivity alone was not associated with any increase in the inflammatory markers in any of the two groups . However, in CHD patients seropositivity to both agents predicted higher levels of TNFalpha (P=0.041), CRP (P=0.037) and fibrinogen (P=0.001) compared to double seronegativity . We conclude that C . pneumoniae LPS seropositivity may contribute to increased vascular inflammation in CHD patients, possibly even more pronounced when present in combination with H . pylori seropositivity. Vet Microbiol, 2002 Aug 2, 88(1), 59 - 74 Characterization of the in vitro adhesion of Actinobacillus pleuropneumoniae to swine alveolar epithelial cells; Van Overbeke I et al.; Actinobacillus pleuropneumoniae biovar 1 serotypes 2, 5a, 9 and 10 strains were tested for their ability to adhere to alveolar epithelial cells in culture . For the serotypes 5a, 9 and 10 strains, optimal adherence was observed after growth of bacterial cells in a NAD-restricted medium (0.001% NAD) . This condition was also associated with the expression of a 55 kDa outer membrane protein (OMP) and of fimbriae . For the serotype 2 strain, adherence and expression of fimbriae and a 55 kDa OMP was less influenced by the growth conditions . The N-terminal amino acid sequence of the 55 kDa OMP had no homology with any known sequence, suggesting that it is an as yet unknown protein . Adherence capabilities were significantly reduced following treatment of the bacteria with proteolytic enzymes or heat . These findings suggest that proteins are involved in adhesion . The hydrophobic bond-breaking agent tetramethylurea was unable to inhibit the adherence of A . pleuropneumoniae to alveolar epithelial cells . Treatment of the bacteria with sodium metaperiodate resulted in lower adhesion scores for the serotypes 2 and 9 strains but the inhibition of adhesion was clearly lower than after treatment with proteolytic enzymes . This indicates that, besides proteins, carbohydrates might also be involved in adhesion of A . pleuropneumoniae to alveolar epithelial cells . The finding that inhibition of adhesion was very high when bacteria were treated with a combination of sodium metaperiodate and pronase also suggests that more than one adhesin is involved. Gene, 2002 Jun 12, 292(1-2), 245 - 59 Structure, expression, and functional characterization of the mouse CLP-1 gene; Huang F et al.; Mouse CLP-1, a potential cardiac transcriptional regulatory factor, is encoded by a single copy gene lacking introns that is expressed into two mRNAs via alternative polyadenylation . Both mRNAs encode the same 41 kDa protein, a novel protein that is 85.3% homologous with a human homologue called HIS1 . Mouse CLP-1 is widely expressed in a number of tissues as well as in early development and is localized to the nucleus . The CLP-1 gene promoter is active in different cell types and sequence analysis shows a number of potential binding sites for cardiogenic transcription factors such as Nkx2.5 and GATA-4, indicating a potential role in development . CLP-1 appears to "squelch" the cardiac MLC-2v promoter in a concentration-dependent manner in cardiac but not other cell types, suggesting that CLP-1 may be interacting with a cardiac-specific factor to regulate cardiac MLC-2v expression . The overall expression pattern of CLP-1 is similar to that of LCR-F1 and Oct-1, two widely expressed transcription factors that also play specific roles in the transcription of cell-specific genes . CLP-1 may be a transcriptional mediator capable of interacting with and potentiating cell-specific transcription factors. Biochemistry, 2002 Jul 23, 41(29), 9237 - 47 New insights into the heme cavity structure of catalase-peroxidase: a spectroscopic approach to the recombinant synechocystis enzyme and selected distal cavity mutants; Heering HA et al.; Catalase-peroxidases (KatGs) are heme peroxidases with homology to yeast cytochrome cperoxidase (CCP) and plant ascorbate peroxidases (APXs) . KatGs exhibit a peroxidase activity of broad specificity and a high catalase activity, which strongly depends on the presence of a distal Trp as part of the conserved amino acid triad Arg-Trp-His . By contrast, both CCP and APX do not have a substantial catalase activity despite the presence of the same triad . Thus, to elucidate structure-function relationships of catalase-peroxidases (for which no crystal structure is available at the moment), we performed UV-Vis and resonance Raman studies of recombinant wild-type KatG from the cyanobacterium SynechocystisPCC 6803 and the distal side variants (His123-->Gln, Glu; Arg119-->Ala, Asn; Trp122-->Phe, Ala) . The distal cavity of KatG is very similar to that of the other class I peroxidases . A H-bond network involving water molecules and the distal Trp, Arg, and His is present, which connects the distal and proximal sides of the heme pocket . However, distal mutation not only affects the heme Fe coordination state and perturbs the proximal Fe-Im bond, as previously observed for other peroxidases, but also alters the stability of the heme architecture . The charge of the distal residues appears particularly important for maintaining the heme architecture . Moreover, the Trp plays a significant role in the distal H-bonding, much more pronounced than in CCP . The relevance of these findings for the catalase activity of KatG is discussed in light of the complete loss of catalase activity in the distal Trp mutants. Infect Immun, 2002 Aug, 70(8), 4581 - 90 Helicobacter pylori-induced activation of human endothelial cells; Innocenti M et al.; Helicobacter pylori infection causes active chronic inflammation with a continuous recruitment of neutrophils to the inflamed gastric mucosa . To evaluate the role of endothelial cells in this process, we have examined adhesion molecule expression and chemokine and cytokine production from human umbilical vein endothelial cells stimulated with well-characterized H . pylori strains as well as purified proteins . Our results indicate that endothelial cells actively contribute to neutrophil recruitment, since stimulation with H . pylori bacteria induced upregulation of the adhesion molecules VCAM-1, ICAM-1, and E-selectin as well as the chemokines interleukin 8 (IL-8) and growth-related oncogene alpha (GRO-alpha) and the cytokine IL-6 . However, there were large variations in the ability of the different H . pylori strains to stimulate endothelial cells . These interstrain variations were seen irrespective of whether the strains had been isolated from patients with duodenal ulcer disease or asymptomatic carriers and were not solely related to the expression of known virulence factors, such as the cytotoxin-associated gene pathogenicity island, vacuolating toxin A, and Lewis blood group antigens . In addition, one or several unidentified proteins which act via NF-kappaB activation seem to induce endothelial cell activation . In conclusion, human endothelial cells produce neutrophil-recruiting factors and show increased adhesion molecule expression after stimulation with certain H . pylori strains . These effects probably contribute to the continuous recruitment of neutrophils to H . pylori-infected gastric mucosa and may also contribute to tissue damage and ulcer formation. Infect Immun, 2002 Aug, 70(8), 4501 - 9 Dissemination of Mycobacterium tuberculosis is influenced by host factors and precedes the initiation of T-cell immunity; Chackerian AA et al.; We report that dissemination of Mycobacterium tuberculosis in the mouse is under host control and precedes the initiation of T-cell immunity . Nine to eleven days after aerosol inoculation, M . tuberculosis disseminates to the pulmonary lymph nodes (LN), where M . tuberculosis-specific T cells are detected 2 to 3 days thereafter . This indicates that the initial spread of bacteria occurs via lymphatic drainage and that the acquired T-cell immune response is generated in the draining LN . Dissemination to peripheral sites, such as the spleen and the liver, occurs 11 to 14 days postinfection and is followed by the appearance of M . tuberculosis-specific T cells in the lung and the spleen . In all cases studied, dissemination to the LN or the spleen preceded activation of M . tuberculosis-specific T cells in that organ . Interestingly, bacteria disseminate earlier from the lungs of resistant C57BL/6 mice than from the lungs of susceptible C3H mice, and consequently, C57BL/6 mice generate an immune response to M . tuberculosis sooner than C3H mice generate an immune response . Thus, instead of spreading infection, early dissemination of M . tuberculosis may aid in the initiation of an appropriate and timely immune response . We hypothesize that this early initiation of immunity following inoculation with M . tuberculosis may contribute to the superior resistance of C57BL/6 mice. J Food Prot, 2002 Jul, 65(7), 1200 - 6 A review of aerobic and psychrotrophic plate count procedures for fresh meat and poultry products; Jay JM; This is a review of reports that employed aerobic plate counts on fresh meat and poultry products since 1985; it lists synopses of 100 applications . A total of 15 different plating media were used, with 48 (48%) being either plate count agar (PCA) or tryptone glucose yeast extract agar . The temperature-time relations ranged from a low temperature of 20 degrees C for 120 h to 37 degrees C for 24 h . Some 29 different temperature-time combinations were used among the total of 109, with 21 (19.3%) being 35 degrees C/48 h, followed by 12 (11.0%) at 32 degrees C/48 h, 11 (10.1%) at 25 degrees C/48 h, and 9 (8.3%) at 25 degrees C/72 h . Fifty-four (49.5%) plate count applications employed incubation temperatures of 30 degrees C and below . From the 26 reports that employed psychrotrophic counts, 16 (61.5%) used PCA; 18 different temperature-time combinations were used, with 7 degrees C/10 d employed by only four . Twenty-one (80.8%) employed an incubation temperature at or <10 degrees C, and five employed an incubation temperature >10 degrees C . There is a serious need for some consensus on methodologies for aerobic and psychrotrophic counts on fresh meat and poultry products. Equine Vet J, 2002 Jul, 34(4), 405 - 10 Intradermal skin testing in Icelandic horses in Austria; Kolm-Stark G et al.; Icelandic horses in Austria are commonly affected by an allergic inflammatory skin disease recurring during the summer seasons, which shares characteristic features with Culicoides hypersensitivity . However, the causative agents have not yet been identified . Therefore, intradermal skin testing (IDST) with a standardised extract of Culicoides variipennis and 21 other allergens relevant within Austria was performed in 81 Icelandic horses . All horses included into the study were treated regularly with ivermectin and had no history of administration of anti-inflammatory drugs . Forty-three of these horses were affected by summer seasonal recurrent dermatitis (SSRD) . No history or signs of any other disease were evident in any horse . Pruritic dermatitis due to ectoparasites, bacteria and dermatophytes were ruled out by means of fungal culture, skin scraping and biopsy . Culicoides variipennis antigens evoked a positive cutaneous reaction in 1 of 38 normal and 3 of 43 SSRD horses at the proposed dilution of 1:50,000 or 1:25,000, and in 24 of 38 normal and 13 of 43 SSRD horses at a dilution of 1:10,000 . Furthermore, no significant differences in onset or intensity of skin reactions to the 21 other allergens, including pollens, moulds, mites and insects, except deerfly and horsefly, were obvious between the 2 groups . Efficiency (percentage of correct results) for the used antigens in the skin test was 0.47-0.60 . Maximal sensitivity was 0.51 . Altogether, 38 of 43 SSRD horses and 28 of 38 normal horses were positive 4 h after allergen administration . The divergence between IDST results and manifestation of clinical signs found in this study underlines the difficulties associated with establishing a skin test protocol in horses within a geographic area . Whether the outcome of this study would have been influenced significantly by using Culicoides spp . present in Austria has to be clarified in future research. Med Device Technol, 2002 Jun, 13(5), 20 - 3 Trends in medical filtration; Hogan B; Advances in materials, mould tooling and control systems are offering the industry greater design choices in filtration as well as the potential to reduce manufacturing costs . This article describes what is possible. J Med Virol, 2002 Aug, 67(4), 603 - 7 Foodborne outbreak caused by a Norwalk-like virus in India; Girish R et al.; An outbreak of acute gastroenteritis occurred in the nurses' hostel of a civil hospital in Delhi, after a farewell party involving 130 nurses and some of the housekeeping staff . All affected persons had eaten salad sandwiches at the party . Stool samples were collected from six of these patients on the second day of infection . All six samples, when tested for the presence of common bacteria, parasites, and rotavirus, were found to be negative . The clinical features of this outbreak matched the criteria set for outbreaks caused by Norwalk-like viruses (NLVs) . Reverse transcription-polymerase chain reaction (RT-PCR) was carried out on these six samples, using primers from the RNA-dependent RNA polymerase (RdRp) gene of NLVs . Immunoelectron microscopy was carried out on two of the samples, using convalescent phase serum . All six samples were positive for genogroup (GG) II NLVs by RT-nested PCR . Aggregates of 32-nm viral particles were visualized by immunoelectron microscopy in one of the two samples . Sequencing of the RdRp gene was done on amplicons from three samples; phylogenetic analysis placed the isolates NDV/1999 in a Toronto virus cluster of GG II NLVs . This is the first report of a food-borne outbreak attributable to NLVs from India . Biotechnol Bioeng, 2002 Aug 5, 79(3), 314 - 22 Conversion and toxicity characteristics of formaldehyde in acetoclastic methanogenic sludge; Gonzalez-Gil G et al.; An unadapted mixed methanogenic sludge transformed formaldehyde into methanol and formate . The methanol to formate ratio obtained was 1:1 . Formaldehyde conversion proceeded without any lag phase, suggesting the constitutive character of the formaldehyde conversion enzymes involved . Because the rate of formaldehyde conversion declined at increased formaldehyde additions, we hypothesized that some enzymes and/or cofactors might become denatured as a result of the excess of formaldehyde . Furthermore, formaldehyde was found to be toxic to acetoclastic methanogenesis in a dual character . Formaldehyde toxicity was partly reversible because once the formaldehyde concentration was extremely low or virtually removed from the system, the methane production rate was partially recovered . Because the degree of this recovery was not complete, we conclude that formaldehyde toxicity was partly irreversible as well . The irreversible toxicity likely can be attributed to biomass formaldehyde-related decay . Independent of the mode of formaldehyde addition (i.e., slug or continuous), the irreversible toxicity was dependent on the total amount of formaldehyde added to the system . This finding suggests that to treat formaldehyde-containing waste streams, a balance between formaldehyde-related decay and biomass growth should be attained . Arch Microbiol, 2002 Aug, 178(2), 85 - 93 Epub 2002 Jun 14. Genetics and control of CO(2) assimilation in the chemoautotroph Ralstonia eutropha; Bowien B et al.; The nutritional versatility of facultative autotrophs requires efficient overall control of their metabolism . Most of these organisms are Proteobacteria that assimilate CO(2) via the highly energy-demanding Calvin-Benson-Bassham reductive pentose-phosphate cycle . The enzymes of the cycle are encoded by cbb genes organized in cbb operons differing in size and composition, although conserved features are apparent . Transcription of the operons, which may form regulons, is strictly controlled, being induced during autotrophic but repressed to varying extents during heterotrophic growth of the bacteria . The chemoautotroph Ralstonia eutropha is one of the organisms studied extensively for the mechanisms involved in the expression of cbb gene systems . CbbR is a LysR-type transcriptional regulator and the key activator protein of cbb operons . The cbbR gene is typically located adjacent and in divergent orientation to its cognate operon . The activating function of CbbR seems to be modulated by metabolites signaling the nutritional state of the cell to the cbb system . Phosphoenolpyruvate is such a signal metabolite acting as a negative effector of R . eutropha CbbR, whereas NADPH has been proposed to be a coactivator of the protein in two other chemoautotrophs, Xanthobacter flavus and Hydrogenophilus thermoluteolus . There is evidence for the participation of additional regulators in cbb control . In the photoautotrophs Rhodobacter capsulatus and Rhodobacter sphaeroides, response regulator RegA of the global two-component signal transduction system RegBA serves this function . It is conceivable that specific variants of cbb control systems have evolved to ensure their optimal integration into regulatory networks operating in the diverse autotrophs characterized by different metabolic capabilities. Plant Physiol, 2002 Jul, 129(3), 1216 - 21 Effect of short-term N(2) deficiency on expression of the ureide pathway in cowpea root nodules; Smith PM et al.; Root systems of 28-d-old cowpea (Vigna unguiculata L . Walp cv Vita 3: Bradyrhizobium sp . strain CB756) plants bearing nitrogen-fixing nodules in sand culture were exposed to an atmosphere of Ar:O(2) (80:20, v/v) for 48 h and then returned to air . Root systems of control plants were maintained in air throughout . Nodules were harvested at the same times in control and Ar:O(2)-treated root systems . Activities of two enzymes of de novo purine synthesis, glycinamide ribonucleotide transformylase (GART; EC 2.1.2.2), aminoimidazole ribonucleotide synthetase (AIRS; EC 6.3.3.1), uricase (EC 1.7.3.3), and phosphoenolpyruvate carboxylase (PEPC; EC 4.1.1.31) were measured together with the protein level of each using immune-specific polyclonal antibodies . AIRS activity and protein both declined to very low levels within 6 h in Ar:O(2) together with a decline in transcript level of pur5, the encoding gene . GART activity, protein, and transcript (pur3) levels were relatively stable . Uricase activity declined in Ar:O(2) as rapidly as AIRS activity but the protein was stable . PEPC activity showed evidence of increased sensitivity to inhibition by malate but the protein level was stable . The data indicate that the flux of fixed N from bacteroids (N(2)-fixing nodule bacteria) is in some way associated with transcriptional control over pur5 and possibly also catabolism of AIRS protein . In contrast, there is limited posttranslational control over GART and PEPC and close posttranslational control over uricase activity . The significance of these different levels of regulation is discussed in relation to the overall control of enhanced expression of plant enzymes in the cowpea symbiosis. Trends Mol Med, 2002 Jul, 8(7), 355 - 8 Blood-bank testing for infectious diseases: how safe is blood transfusion? Strong DM, Katz L. Remarkable progress has been made in transfusion safety from infection over the past three decades . Donor deferrals for at-risk behaviors, the introduction of more-sensitive viral-screening assays and the recent introduction of nucleic-acid amplification technology have nearly eliminated transmission of HIV and hepatitis C virus (HCV) by blood transfusion in North America . Nevertheless, risks of other infectious agents for which such robust screening tools have not been developed, such as bacteria and parasites, still remain . As a result of these successes, the non-infectious risks such as misidentification of patients and inadequate and inappropriate transfusion have become the primary sources of transfusion risk. J Periodontal Res, 2002 Jun, 37(3), 210 - 4 Treatment with SRL172 (heat-killed Mycobacterium vaccae) inhibits progression of established experimental periodontal disease in Wistar rats; Brelvik T et al.; Periodontal disease is accompanied by a change in the periodontal bacterial flora, and an increase in Th2 cytokine expression . Therefore, preparations that can down-regulate Th2 responses and increase Th1 responses to bacteria may have therapeutic effects . SRL172, a preparation of heat-killed Mycobacterium vaccae (NCTC11659), has been shown to have these properties in man and animals and, in a previous study, a single subcutaneous injection of this material 13 days before application of ligatures to the right second molars of Wistar rats strikingly reduced subsequent destruction of the tooth-supporting tissues . The same material has therefore been tested in a therapeutic protocol, in rats with ongoing periodontal disease . A silk ligature was placed round the maxillary right second molar in the gingival sulcus on day 0 . This was removed after two weeks, and the animals immediately received 0.1 mg SRL172 s.c . or saline . One week later (i.e . at three weeks), a boost of 1.0 mg SRL172 or saline was given . Animals were sacrificed at eight weeks (i.e . five weeks after the booster dose) . Treatment of ongoing periodontal disease with SRL172 significantly reduced fibre loss (p = 0.046 by histometry) and bone loss (p = 0.0008 by radiography) on the ligatured side, and reduced fibre loss (p = 0.0086) on the control side . Thus SRL172 is an effective treatment in this model . As SRL172 has undergone extensive safety testing in man, these results justify clinical studies in periodontal disease, and such studies are now planned. J Periodontal Res, 2002 Jun, 37(3), 177 - 83 Upregulation of co-stimulatory molecule expression and dendritic cell marker (CD83) on B cells in periodontal disease; Mahanonda R et al.; T cells and their cytokines are well known for their important role in the pathogenesis of periodontitis . To date, the role of antigen presenting cells (APCs), which are known to be critical in the regulation of T cell response, has been poorly investigated in periodontitis . In this study, we analyzed the expression of co-stimulatory molecules (CD80 and CD86) and CD83, which is a marker of mature dendritic cells, on gingival cells that were isolated from severe periodontitis tissues, with the use of flow cytometry . Significant upregulation of CD86 and CD83 expression was detected in periodontitis lesions, and most of this occurred on B cells . In vitro peripheral blood mononuclear cell cultures showed that stimulation with different periodontopathic bacteria, that included Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, Prevotella intermedia, and Actinomyces viscosus, upregulated both CD86 and CD83 expression on B cells . Therefore, the presence of plaque bacteria may be responsible for the enhanced expression seen in vivo on gingival B cells . APC function by bacterial-activated B cells was further investigated using allogeneic mixed leukocyte reactions . After 24 h culture with either A . actinomycetemcomitans or P . gingivalis, these activated B cells performed as potent APCs in mixed leukocyte reactions, and they stimulated T cells to produce high levels of gamma interferon and minimal interleukin-5 . In conclusion, periodontopathic bacterial-induced B cell activation with upregulation of CD86 and CD83 may be associated with enhanced APC function . The results of this study suggest, therefore, that infiltrated gingival B cells have a possible role as APCs in the regulation and maintenance of local T cell response in periodontitis. Ground Water, 2002 Jul-Aug, 40(4), 416 - 24 Localized thermal anomalies in haloclines of coastal Yucatan sinkholes; Stoessell RK et al.; A temperature spike is reported in the haloclines of three Yucatan sinkholes along a 1 km NW-SE transect from 5 to 4 km inland from the Caribbean coast . The temperature spike decreases in magnitude from 3.5 degrees C to 0.2 degrees C, approaching the coast . The anomaly does not vary diurnally and does not extend down into the underlying sea water . These conditions are inconsistent with explanations such as radiation absorption within the halocline, in situ microbially mediated sulfate reduction within the halocline and the underlying sea water, and sulfide oxidation by photosynthetic purple and green bacteria within the halocline . One explanation consistent with the shape and halocline location of the temperature spike involves a localized sea water convection cell operating near the coast . Cold sea water from the Caribbean Sea enters the coastal limestone at depths of a few hundred meters and heats up because of the geothermal gradient, buoyantly rising in vertical fractures within the unconfined aquifer . Blocked by the less dense fresh water, the movement stops in the halocline where the warm sea water mixes with brackish water . The convection cycle would be completed by the coastward movement of cooling brackish water . The observed temperature anomalies could possibly be snapshots of this warm layer moving coastward. Expert Rev Anticancer Ther, 2002 Jun, 2(3), 309 - 21 Gene therapy for prostate cancer; Gdor Y et al.; Prostate cancer is the most common noncutaneous cancer in man . When confined to the prostate it can be cured by radical prostatectomy or irradiation therapy . However, there are no curative therapies for locally advanced, recurrent or metastatic disease . Prostate cancer gene therapy has recently transition from preclinical studies to clinical trials with the goal of developing novel treatments for prostate cancer . The greatest challenge in treating advanced prostate cancer is therapeutic access to and the elimination of metastases . This review details two aspects of prostate cancer gene therapy, the types of delivery systems under development and specific categories of therapeutic genes available with an emphasis on the mechanism of action of specific gene therapy strategies. Proteomics, 2002 Jun, 2(6), 775 - 83 Immunogenic proteins of Helicobacter pullorum, Helicobacter bilis and Helicobacter hepaticus identified by two-dimensional gel electrophoresis and immunoblotting; Kornilovs'ka I et al.; The ecological niches occupied by various species of Helicobacter are not yet known and the full spectrum of diseases associated with Helicobacter infections are not yet defined . Since these fastidious microaerofilic bacteria require special growth conditions new and improved molecular and serologic diagnostic methods have been developed to increase our understanding of their pathogenesis and virulence characteristics . Immunogenic cell surface proteins of Helicobacter pullorum, Helicobacter bilis, and Helicobacter hepaticus were characterised by proteomic techniques using two-dimensional electrophoresis and immunoblotting with antisera from immunised rabbits . Cross-reactivity between the three Helicobacter species were analysed after a four-step cross-absorption experiment . For H . pullorum, H . bilis and H . hepaticus 21, 13 and 27 specific immunogenic proteins, respectively, were identified . These proteins could be of important sero-diagnostic value for analyses of sera from humans, laboratory animals and for the veterinarian field. Environ Toxicol, 2002, 17(3), 149 - 59 Biomarker study of a municipal effluent dispersion plume in two species of freshwater mussels; Gagne F et al.; The toxicological effects of a primary-treated municipal effluent plume were investigated in two species of freshwater mussels, Elliptio complanata and Dreissena polymorpha, exposed for 62 days at sites upstream and downstream of an effluent outfall in the St . Lawrence River (Quebec, Canada) . Levels of metallothioneins (MT), cytochrome P4501A1 activity, DNA damage, total lipids, relative levels of vitellins, and phagocytic activity (in E . complanata hemocytes) were determined after the exposure period . A parallel analysis measured heavy metals and coprostanol in mussel tissues . The results show that significant levels of coprostanol and some metals (specifically, Cu, Hg, Sb, Se, and Zn) had accumulated in mussels caged 5 km downstream of the effluent plume . Mixed-function oxidase activity, MT in gills, total lipids, DNA damage (in D . polymorpha only), and total hemolymph bacteria (in E . complanata only) had increased in these mussels, while levels of total cadmium (Cd), MT in digestive glands or whole soft tissues, phagocytic activity, and DNA damage in the digestive gland (in E . complanata only) were diminished . The exposure of mussels to surface waters contaminated by a municipal effluent led to many stress responses, depending on both the tissues and the species being examined . Microsc Res Tech, 2002 Jun 15, 57(6), 491 - 7 Are thrombocytes and platelets true phagocytes? Meseguer J, Esteban MA, Rodriguez A. Thrombocytes and platelets, beyond their primary function in hemostasis, seem to play an active role in inflammation . As regards their phagocytic ability, the results to date are confusing, incomplete, and somewhat contradictory . Whereas the interaction of avian thrombocytes or mammalian platelets with bacteria both in vitro and in vivo has received wide attention, almost no information exists on the topic in "lower" vertebrates . The aim of this work is to review the available information on the phagocytic properties of thrombocytes and platelets . Particular attention is payed to the ontogeny of these cells, the soluble factors involved in the inflammatory process derived from them, and their interaction with particulate material, mainly with bacteria . J Cell Biochem, 2002, 86(2), 277 - 89 Sox9 transactivation and testicular expression of a novel human gene, KIAA0800; Zhao LJ et al.; The Sry and Sox9 sex-determination factors initiate and promote testis differentiation by gene transactivation through similar promoter elements . However, knowledge is limited concerning what genes are regulated by Sry/Sox9 in the testis . Identification and characterization of Sry/Sox9-regulated genes are critical for understanding sexual differentiation . We now demonstrate that a novel human gene, KIAA0800, is preferentially expressed in the testis and is transactivated by Sox9 . The KIAA0800 promoter is repressed by an upstream element involving a polyT track and two Alu repeats . Two specific Sox9-bindings sites have been identified in the KIAA0800 promoter by using DNaseI footprinting assays and gel electrophoretic mobility shift assays . Sox9 transactivation of the KIAA0800 promoter appears to be exerted mainly through the relief of promoter repression . Genes homologous to the human KIAA0800 exist in organisms with differentiated sex tissues including mouse, Drosophila, and C . elegans, but not in unicellular organisms, including yeast and bacteria . Further, our recent sequence analysis shows that KIAA0800 protein is 97% identical between human and mouse . Thus, KIAA0800 is a novel Sox9-activated gene that is evolutionarily conserved and potentially involved in sexual differentiation . Eur J Clin Microbiol Infect Dis, 2002 Jun, 21(6), 455 - 60 Epub 2002 Jun 14. Evaluation of a new commercial assay for diagnosis of pulmonary and nonpulmonary tuberculosis; Johansen IS et al.; A new commercial assay for the diagnosis of tuberculosis, the BDProbeTec ET Direct Detection assay (Becton Dickinson, USA), was evaluated using 351 respiratory and 372 nonrespiratory specimens . The results were compared to detection of Mycobacterium tuberculosis complex (MTC) by conventional culture . Among the 351 respiratory specimens, MTC bacteria were identified in 150, of which 85 were positive by both microscopy and the assay . Sixty-five specimens culture positive for MTC were microscopy negative; of these, 39 were positive in the assay . All 26 specimens culture positive for nontuberculous mycobacteria (NTM) were negative by the assay . Of 175 specimens culture negative for MTC, 3 were falsely positive by the assay and 1 yielded inhibition . The overall sensitivity and specificity values were 82.7% and 98.5%, respectively . The sensitivity for microscopy-positive and -negative respiratory specimens was 100% and 60%, respectively . After correction for discrepancies, the specificity was 99% compared with notification data . The BDProbeTec ET assay detected 66 of 67 microscopy-positive and 50 of 125 microscopy-negative nonrespiratory specimens . The result for one specimen was inconclusive . All nine specimens containing NTM were negative by the assay . Of 171 specimens culture negative for MTC, 6 were falsely positive by the assay . The overall sensitivity and specificity values obtained with nonrespiratory specimens were 60.7% and 96.7%, respectively . After examining discrepancies by reviewing the patients' histories, the specificity was 98.9% . The sensitivity was 98.5% in microscopy-positive specimens and 40.3% in microscopy-negative specimens . The overall inhibition rate was 0.3% . The BDProbeTec ET assay is a fast, effective, and user-friendly system that can be used for rapid detection of MTC bacteria in respiratory and microscopy-positive nonrespiratory specimens as an important supplement to smear and culture. Mol Genet Genomics, 2002 Jun, 267(4), 526 - 35 Epub 2002 May 29. Identification and genomic organization of gene loci negatively controlled by the virulence regulatory BvgAS two-component system in Bordetella bronchiseptica; Schneider B et al.; The two-component system BvgAS positively controls transcription of the virulence genes of Bordetella pertussis and B . bronchiseptica, which include several genes for toxins and adhesins . On the other hand, the BvgAS system negatively controls the expression of a poorly characterized set of genes, the so-called virulence repressed ( vrg) genes . To investigate the function of this group of genes and their relationship to virulence, we identified several novel vrg genes of B . bronchiseptica via the generation of transcriptional fusions with gfp (the ORF encoding Green Fluorescent Protein) by transposon mutagenesis . Expression of all of the vrg genes was enhanced under phenotype-modulating growth conditions and in phase variants, demonstrating that their transcription is indeed controlled by the BvgAS system . In addition, transcription of most of these new vrg genes was found to be affected by the growth phase of the bacteria, with maximal expression being observed in the late logarithmic or stationary phase . The majority of these genes encode putative metabolic functions involved in redox reactions and amino acid transport . Interestingly, several vrg genes of B . bronchiseptica are not expressed or have been lost in B . pertussis, indicating that, possibly as a consequence of its adaptation to a single host organism, many vrg genes of B . pertussis are gradually decaying. Appl Microbiol Biotechnol, 2002 Jul, 59(2-3), 231 - 8 Epub 2002 May 16. Introduction of the carbohydrate-activated promoter P(malK) for recombinant protein production; Bostrom M et al.; A production protocol for the use of the malK promoter was established . The protocol includes two phases: an initial fed-batch phase on glucose to reach a high cell density and a fed-batch phase on maltose for production of the desired recombinant protein . It is suggested that this cultivation scheme could be used for all promoters that are catabolite repressed by glucose and where growth and production need to be separated . The specific feature of this system is shown by its ability to control the rate of synthesis of the product protein, ss-galactosidase . In the production phase with a constant feed or an exponential feeding of 0.1 h(-1) it took 4 h longer to reach the maximum specific production rate than with the higher dilution rates of 0.25 h(-1) and 0.4 h(-1), respectively . In the above experiments a dilution rate of 0.3 h(-1) in the growth phase was used . The volumetric production of this system could furthermore be extended to 40 h . All protocol procedures so far tested resulted in the same maximum production rate, but reached in different lengths of time . It is argued that this system is particularly well suited for the production of proteins that have a complex structure and/or need to be produced in a soluble form or to be exported to the periplasm. FEMS Immunol Med Microbiol, 2002 Jul 12, 33(3), 191 - 200 Major histocompatibility complex class I and II expression on macrophages containing a virulent strain of Brucella abortus measured using green fluorescent protein-expressing brucellae and flow cytometry; Murphy E et al.; Immune responses appropriate for control of an intracellular pathogen are generated in mice infected with Brucella abortus, shown by the ability of T cells to adoptively transfer resistance to naive mice . The infection nevertheless persists for months . It was hypothesized that one factor in maintaining the infection despite the presence of immune T cells was suboptimal expression of major histocompatibility complex (MHC) molecules on macrophages containing brucellae . This would allow B . abortus to elude detection by the host's immune system . To test this, B . abortus organisms expressing green fluorescent protein (GFP-Brucella) were constructed and three-color flow cytometry used to evaluate MHC expression on macrophages following in vitro or in vivo infection . When infected in vitro, the levels of MHC class I and class II expression on J774 macrophages containing GFP-Brucella were the same or higher than on macrophages without GFP-Brucella in the same cultures . Similarly, the MHC expression was higher on GFP(+) peritoneal exudate cells following infection or phagocytosis of heat-killed GFP-Brucella than it was on uninfected peritoneal exudate cells . Following in vivo infection of mice the level of MHC class I and II expression on GFP(+) cells in their spleens (the main site of infection) also tended to be as high as or higher than that on the GFP-negative cells . The only in vivo GFP(+) cells that showed a decreased MHC expression was a population of splenic Mac1(+) cells recovered from interferon-gamma gene-disrupted mice at the time of their death due to an overwhelming number of bacteria per spleen . Overall, it was concluded that decreased MHC expression is not a general principle associated with brucella infection of macrophages and thus not likely to contribute to maintenance of the chronic infection. Crit Rev Microbiol, 2002, 28(2), 79 - 122 Ribonucleases from T2 family; Deshpande RA et al.; Ribonucleases are ubiquitous in distribution . Ribonucleases that hydrolyse RNA to 3' mononucleotides via 2', 3' cyclic nucleotides are classified into three groups, RNase A, RNase T1, and RNase T2 families . Apart from salvage of cellular or extracellular RNAs, RNases participate in vital cellular functions such as DNA replication, transcription and RNA processing, splicing and editing, and control of translation by determining the turnover of RNA . T2 family RNases have been implicated in nutrition, phosphate remobilization, self-incompatibility, senescence, and defense against pathogens . They are important analytical enzymes and have been exploited for the structural determination of RNAs . Although considerable information is available on RNase A and T1 family RNases, less information is available on RNases from T2 family except RNase Rh from Rhizopus niveus and RNase LE from tomato . However, during the last few years, the primary structure, active site nature based on sequence homology, and probable mechanism of action have been postulated for some of these enzymes . RNases of T2 family, their occurrence, purification, characteristics, biological role, and applications have been reviewed. Sci Total Environ, 2002 Jul 3, 293(1-3), 219 - 31 Bioaerosol exposure during refuse collection: results of field studies in the real-life situation; Neumann HD et al.; To determine the bioaerosol exposure of refuse collectors, field measurements were performed under real working conditions within the framework of a research project . Influencing variables such as different types of refuse, community structure, collection interval and season were taken into account . Overall, 1612 samples were taken in towns of Westfalia, Germany . With workplace levels on a scale of 10(3) to less than 10(4) CFU/m3 for the loader, the results show a surprisingly low total fungi concentration in comparison with earlier studies . Total bacteria concentrations, in contrast, were largely on a scale of 10(4) CFU/m3, with 10(5) CFU/m3 being registered sporadically, especially in apartment-block districts . Endotoxin levels were high especially in the summer months, occasionally reaching values of more than 50 EU/m3, whereas they were normally below 10 EU/m3 in autumn and winter . Inside the cab, the exposure level for the entire spectrum was at least one power of ten lower . The factors believed to account primarily for the low total fungi concentration were workplace hygiene, the prevailing 1-week collection interval, and the low in-process exposure time resulting from the effective deployment of automatic lifting devices . In contrast, the type of refuse was not found to have a significant influence. Curr Gene Ther, 2001 May, 1(1), 53 - 100 Genetic vaccination for the active immunotherapy of cancer; Bronte V; Molecular biology techniques have given novel impetus to the immunotherapy of cancer because they have catalyzed the identification of several potential tumor antigens, and permitted the generation of vectors for the delivery of genetic material encoding these antigens . Vaccines can be defined "genetic" when the antigen they enclose is present as DNA or RNA . Microrganisms used as vectors can deliver the genetic information, but naked nucleic acids have also been shown to be effective immunogens thanks to built-in adjuvants that activate professional antigen presenting cells . Although gene-based cancer vaccines have been tested in mouse models and selected for pilot clinical trials, enthusiasm has somewhat waned due to an apparently major drawback of cancer vaccination: tumor antigens are weak, and therefore fail to stimulate a sterilizing immune response in tumor-bearing patients . Mouse studies, however, have shown that cancer vaccines are extremely efficacious in establishing a state of active immunosurvellance against tumor growth . This review reconsiders the findings emerging from preclinical studies in the context of our current knowledge of the cellular and molecular bases of the immune responses to vaccines, in an attempt to approach critically the use of genetic vaccination for the treatment of cancer. Water Res, 2002 May, 36(9), 2225 - 32 Treatment of domestic sewage in a two-step anaerobic filter/anaerobic hybrid system at low temperature; Elmitwalli TA et al.; The treatment of domestic sewage at low temperature of 13 degrees C was investigated in a two-step system consisting of an anaerobic filter (AF) +an anaerobic hybrid (AH) reactor operated at different hydraulic retention times (HRTs) . The AF reactor was efficient in the removal of suspended COD, viz . 81%, 58% and 57% at an HRT of, respectively, 4, 2 and 3 h . For optimisation of the removal of suspended COD and dissolved COD, an HRT of 4 + 4 h is required for the AF + AH system . For additional optimisation of colloidal COD removal, the AH reactor needs an HRT of 8 h . The AF + AH system operated at an HRT of 4 + 8 h at 13 degrees C provided a high removal efficiency for all COD fractions . The achieved total COD removal was as high as 71% which is similar to values found in tropical areas . Moreover, 60% of the removed COD was converted to methane. Br J Cancer, 2002 Jul 15, 87(2), 187 - 93 Establishment and characterisation of six human biliary tract cancer cell lines; Ku JL et al.; Human cell lines established from biliary tract cancers are rare, and only five have been reported previously . We report the characterisation of six new six biliary tract cancer cell lines (designated SNU-245, SNU-308, SNU-478, SNU-869, SNU-1079 and SNU-1196) established from primary tumour samples of Korean patients . The cell lines were isolated from two extrahepatic bile duct cancers (one adenocarcinoma of common bile duct, one hilar bile duct cancer), two adenocarcinomas of ampulla of Vater, one intrahepatic bile duct cancer (cholangiocarcinoma), and one adenocarcinoma of the gall bladder . The cell phenotypes, including the histopathology of the primary tumours and in vitro growth characteristics, were determined . We also performed molecular characterisation, including DNA fingerprinting analysis and abnormalities of K-ras, p15, p16, p53, hMLH1, hMSH2, DPC4, beta-catenin, E-cadherin, hOGG1, STK11, and TGF-betaRII genes by PCR-SSCP and sequencing analysis . In addition, we compared the genetic alterations in tumour cell lines and their corresponding tumour tissues . All lines grew as adherent cells . Population doubling times varied from 48-72 h . The culture success rate was 20% (six out of 30 attempts) . All cell lines showed (i) relatively high viability; (ii) absence of mycoplasma or bacteria contamination; and (iii) genetic heterogeneity by DNA fingerprinting analysis . Among the lines, three lines had p53 mutations; and homozygous deletions in both p16 and p15 genes were found three and three lines, respectively; one line had a heterozygous missense mutation in hMLH1; E-cadherin gene was hypermethylated in two lines . Since the establishment of biliary tract cancer cell lines has been rarely reported in the literature, these newly established and well characterised biliary tract cancer cell lines would be very useful for studying the biology of biliary tract cancers, particularly those related to hypermethylation of E-cadherin gene in biliary tract cancer. J Mol Evol, 2002 Aug, 55(2), 153 - 60 Updating carbamoylphosphate synthase (CPS) phylogenies: occurrence and phylogenetic identity of archaeal CPS genes; Cammarano P et al.; Among Bacteria the carA and carB genes encoding the small (CarA) and large (CarB) subunits of carbamoylphosphate synthase (CPS) have been lost in certain symbionts (Haemophylus influenzae) and in most obligate intracellular parasites (Chlamydiae, Spirochaetes, Mycoplasmatales, Rickettsiae) having genome sizes in the 0.7- to 1.1-Mb range . Compared to Bacteria, Archaea exhibit a more varied pattern of CPS gene losses and an unusual propensity to incorporate CPS genes derived from both Bacteria and other Archaea . Schematically they fall into three groups . Group 1 taxa (the crenarchaeon Aeropyrum pernix and the euryarchaea Pyrococcus horikoshi and Pyrococcus abyssii) lack CPS genes altogether . Group 2 taxa (comprising Halobacteriales, Thermoplasmales, Methanococcales, Methanomicrobiales, Archaeoglobales) harbor CPS genes whose encoded CarB and CarA subunit proteins are ostensibly bacterial in origin; that is, they are intermixed with bacterial homologues on a phylogeny of concatenated CarA and CarB sequences and are not distinguishable from bacterial sequences after searching for domain-specific amino acid residue positions . Group 3 taxa (the crenarchaea Pyrobaculum aerophilum, Sulfolobus solfataricus, and Sulfolobus tokodaii and the euryarchaeon Pyrococcus furiosus) harbor CPS genes whose encoded proteins appear to be archaeal: consistent with an archaeal origin, the CarA and CarB sequences in this group possess both unique signatures and signatures affiliating them to Eukarya . Based on the topology of the clade comprising the four Group 3 taxa, we argue that CPS genes of P . furiosus (a euryarchaeon) and those of the crenarchaea P . aerophilum, S . solfataricus, and S . tokodaii are of a single type, resulting from the two genes being laterally transferred from a crenarchaeon to P . furiosus. J Mol Evol, 2002 Aug, 55(2), 127 - 37 Comparative molecular evolution of primary (Buchnera) and secondary symbionts of aphids based on two protein-coding genes; Moya A et al.; A+T content, phylogenetic relationships, codon usage, evolutionary rates, and ratio of synonymous versus non-synonymous substitutions have been studied in partial sequences of the atpD and aroQ/pheA genes of primary ( Buchnera) and secondary symbionts of aphids and a set of selected non-symbiotic bacteria, belonging to the five subdivisions of the Proteobacteria . Compared to the homologous genes of the last group, both genes belonging to Buchnera behave in a similar way, showing a higher A+T content, forming a monophyletic group, a loss in codon bias, especially in third base position, an evolutionary acceleration and an increase in the number of non-synonymous substitutions, confirming previous results reported elsewhere for other genes . When available, these properties have been partly observed with the secondary symbionts, but with values that are intermediate between Buchnera and free living Proteobacteria . They show high A+T content, but not as high as Buchnera, a non-solved phylogenetic position between Buchnera, and the other gamma-Proteobacteria, a loss in codon bias, again not as high as in Buchnera and a significant evolutionary acceleration in the case of the three atpD genes, but not when considering aroQ/pheA genes . These results give support to the hypothesis that they are symbionts at different stages of the symbiotic accommodation to the host. Free Radic Biol Med, 2002 Jul 15, 33(2), 220 - 35 The metabolic fate of dietary polyphenols in humans; Rechner AR et al.; Dietary polyphenols are widely considered to contribute to health benefits in humans . However, little is yet known concerning their bioactive forms in vivo and the mechanisms by which they may contribute toward disease prevention . Although many studies are focusing on the bioavailability of polyphenols through studying their uptake and the excretion of their conjugated forms, few are emphasizing the occurrence of metabolites in vivo formed via degradation by the enzymes of colonic bacteria and subsequent absorption . The purpose of this research was to investigate the relationship between biomarkers of the colonic biotransformation of ingested dietary polyphenols and the absorbed conjugated polyphenols . The results show that the majority of the in vivo forms derive from cleavage products of the action of colonic bacterial enzymes and subsequent metabolism in the liver . Those include the glucuronides of 3-hydroxyphenylacetic, homovanillic, vanillic and isoferulic acid as well as 3-(3-methoxy-4-hydroxyphenyl)-propionic, 3-(3-hydroxyphenyl)-propionic acid, and 3-hydroxyhippuric acid . In contrast, intact conjugated polyphenols themselves, such as the glucuronides of quercetin, naringenin and ferulic, p-coumaric, and sinapic acid were detected at much lower levels . The results suggest that consideration should be given to the cleavage products as having a putative role as physiologically relevant bioactive components in vivo. Microbes Infect, 2002 Jul, 4(9), 903 - 14 Toll receptors, CD14, and macrophage activation and deactivation by LPS; Dobrovolskaia MA et al.; This review will focus on the molecular mechanisms of macrophage activation and desensitization by bacterial lipopolysaccharide (LPS) . The most recent advances in the understanding of the function of the LPS receptor complex and its role in the development of the septic shock syndrome and endotoxin tolerance will be discussed. Microbes Infect, 2002 Jul, 4(9), 887 - 95 Genetic approaches to the study of Toll-like receptor function; Takeuchi O et al.; The Toll-like receptor (TLR) family plays a role in sensing invading pathogens . Ten members have been reported to date, seven of which were found to recognize discrete bacterial components . Mouse models lacking each TLR and its signaling molecule are useful tools for the analysis of the innate immune system. Gastroenterology, 2002 Jul, 123(1), 187 - 95 Mechanisms of acid resistance due to the urease system of Helicobacter pylori; Scott DR et al.; BACKGROUND & AIMS: Helicobacter pylori, a neutralophile, uses acid neutralization by urease to combat gastric acidity, allowing gastric colonization . Both acute and chronic acid resistance mechanisms are present . Acute mechanisms of acid adaptation could be due to surface urease, increased inner-membrane urea permeability via UreI, or both . Slower mechanisms may involve increased nickel insertion into apoenzyme, posttranscriptional regulation, or increased enzyme synthesis . The aim of this study was to further define regulation of urease under acidic conditions . METHODS: Surface-bound urease was analyzed by measurement of free and bound urease after centrifugation through a step gradient and by quantitative urease immunostaining of intact and fixed bacteria . Changes in urease synthesis or assembly were determined by incubation of the organisms at pH 5.5 or 7.0 in the absence and presence of chloramphenicol, urea, or nickel chelator and in ureI-positive and -negative organisms . RESULTS: The amount of surface urease was below detection limits with either centrifugation washing or immunostaining . Total bacterial urease activity was increased 3-5-fold by incubation at pH 5.5 in the presence of chloramphenicol but not in nickel-free medium or in ureI knockout organisms . There was also a 3-fold increase in survival of acid shock in acid-adapted organisms . CONCLUSIONS: Surface-bound urease is too low to contribute to acid resistance . Acidic medium pH induces UreI-dependent nickel incorporation into apoenzyme . This augmentation of urease activity increases survival in acid and is part of the gastric colonization strategy of the organism. Cell Microbiol, 2002 Jul, 4(7), 447 - 60 Identification of MEK- and phosphoinositide 3-kinase-dependent signalling as essential events during Chlamydia pneumoniae invasion of HEp2 cells; Coombes BK et al.; The ability of Chlamydia pneumoniae to survive and cause disease is predicated on efficient invasion of cellular hosts . While it is recognized that chlamydial determinants are important for mediating attachment and uptake into non-phagocytic cells, little is known about the bacterial ligands and cellular receptors that facilitate invasion or host cell signal transduction pathways implicated in this process . We used transmission and scanning electron microscopy to demonstrate that attachment of bacteria to host cells induced the appearance of microvilli on host cell membranes . Invasion occurred 30-120 min after cell contact with the subsequent loss of membrane microvilli . Using an epithelial cell infection model, C . pneumoniae invasion caused a rapid and sustained increase in MEK-dependent phosphorylation and activation of ERK1/2, followed by PI 3-kinase-dependent phosphorylation and activation of Akt . Tyrosine phosphorylation of focal adhesion kinase (FAK) preceded its appearance in a complex with the p85 subunit of PI 3-kinase during chlamydial invasion and isoform-specific tyrosine phosphorylation of the docking protein Shc also occurred at the time of attachment and entry of bacteria . Chlamydia entry but not attachment could be abrogated with specific inhibitors of MEK, PI 3-kinase and actin polymerization, demonstrating the importance of these signalling pathways and an intact actin cytoskeleton for C . pneumoniae invasion . These results suggest that activation of specific cell signalling pathways is an essential strategy used by C . pneumoniae to invade epithelial cells. Cell Microbiol, 2002 Jul, 4(7), 411 - 24 Characterization of a secreted Chlamydia protease; Shaw AC et al.; Chlamydiae are obligate intracellular bacteria that are important human pathogens . The Chlamydia genomes contain orthologues to secretion apparatus proteins from other intracellular bacteria, but only a few secreted proteins have been identified . Most likely, effector proteins are secreted in order to promote infection . Effector proteins cannot be identified by motif or similarity searches . As a new strategy for identification of secreted proteins we have compared 2D-PAGE profiles of {35S}-labelled Chlamydia proteins from whole lysates of infected cells to 2D-PAGE profiles of proteins from purified Chlamydia . Several secretion candidates from Chlamydia trachomatis D and Chlamydia pneumoniae were detected by this method . Two protein spots were identified among the candidates . These represent fragments of the 'chlamydial protease- or proteasome-like activity factor' (CPAF) and were clearly present in 2D-PAGE profiles of whole lysates of infected cells but absent from purified Chlamydia . CPAF was recently identified by Zhong and colleagues as a secreted protease which cleaves host cell transcription factors essential for MHC class I and II antigen presentation . The identification of CPAF in this paper verifies the applicability of the described method for the identification of secreted proteins . We extend the findings by Zhong et al . by proteome studies of expression and turnover of C . trachomatis CPAF showing that the degradation of C . trachomatis D CPAF in the host cell is very limited . Furthermore, we show that two fragments of CPAF exist in C . pneumoniae as well as in C . trachomatis. Biochemistry, 2002 Jul 16, 41(28), 8899 - 906 DNA adducts formed from 4-hydroxytamoxifen are more mutagenic than those formed by alpha-acetoxytamoxifen in a shuttle vector target gene replicated in human Ad293 cells; McLuckie KI et al.; The drug tamoxifen, used to treat breast cancer, causes liver cancer in rats and endometrial cancer in women . Tamoxifen forms liver DNA adducts in both short- and long-term dosing of rodents, and DNA adducts have also been reported in tissues of women undergoing tamoxifen therapy . It is not known if the induction of endometrial cancer in women is through these DNA adducts or through the estrogenic nature of the drug . In this study, we have investigated the mutagenicity of two model reactive intermediates of tamoxifen, alpha-acetoxytamoxifen and 4-hydroxytamoxifen quinone methide (4-OHtamQM) . These form the same DNA adducts as those found in tamoxifen-treated rats . The two compounds were used to treat the pSP189 plasmid containing the supF gene, which was replicated in Ad293 cells before being screened in indicator bacteria . Plasmid reacted with 4-OHtamQM was more likely to be mutated (2-7-fold increase) than that reacted with alpha-acetoxytamoxifen, despite having a lower level of DNA damage (12-20-fold less), as assayed by (32)P-postlabeling . The two compounds induced statistically different mutation spectra in the supF gene . The majority of mutations in alpha-acetoxytamoxifen-treated plasmid were GC -->TA transversions while GC-->AT transitions were formed in 4-OHtamQM-treated plasmid . 4-OHTamQM-treated DNA induced a larger proportion of multiple mutations and large deletions compared to alpha-acetoxytamoxifen . Sites of mutational hotspots were observed for both compounds . In conclusion, the quantitatively minor DNA adduct of tamoxifen (dG-N(2)-4-hydroxytamoxifen) is more mutagenic than the major tamoxifen DNA adduct (dG-N(2)-tamoxifen). Biol Sci Space, 2001 Oct, 15 Suppl, S203 - 10 Biological effects of space radiation; Ohnishi T et al.; To determine the effects of space radiation on human health for long-term stays in space, we performed 21 space experiments on radiation biology . Two main characteristics of space are microgravity and space radiation that consists of low dose, chronic exposure at low dose-rates, and heavy particles . Through space experiments, we demonstrated the formation of DNA strand breaks, induced mutations, abnormal cell differentiation and the inducible gene expression of a tumor suppressor gene product, p53, in various kinds of organisms . In addition, we investigated the influence of microgravity on radiation-induced biological effects in in vitro biochemical reaction systems and in vivo cell culture systems of bacteria and lower eukaryotes . We review here the importance of radiation biology studies on space radiation from the viewpoints of human health and biological evolution, from the beginning of life until today, in the context of environmental genotoxic radiation. Microbiology, 2002 Jul, 148(Pt 7), 2019 - 27 Mutational analysis of the role of charged residues in target-cell binding, potency and specificity of the pediocin-like bacteriocin sakacin P; Kazazic M et al.; The significance of charged residues for the target-cell binding, potency and specificity of pediocin-like bacteriocins has been studied by site-directed mutagenesis of sakacin P . Most of the charged residues are located in the N-terminal half, which is thought to mediate the initial binding of these bacteriocins to target cells through electrostatic interaction . All the mutated peptides in which the net positive charge was reduced by one (by replacing a charged residue with threonine) exhibited reduced binding to target cells and a 2-15-fold reduction in potency . The least deleterious of these mutations was the removal of the positive charge in position 8 (H8T) . This mutation was, in fact, less deleterious than the conservative His to Lys mutation, indicating that the positive charge in position 8 per se is not of major importance . Somewhat more deleterious was the removal of positive charges at the N- and C-terminal ends (K1T, K43T) . Most deleterious was the elimination of the positive charge at positions 11 and (but to a lesser extent) 12, demonstrating the importance of the cationic patch in the middle of the N-terminal half of pediocin-like bacteriocins . Mutated peptides in which the net positive charge was increased by one were also constructed . Some of these exhibited increased cell binding and a potency that was the same as (44K, i.e . an extra positive charge at the C-terminus), or somewhat greater (T20K) than, that of sakacin P, whereas others (0K, i.e . an extra positive charge at the N-terminus) had reduced potency . Sakacin P contains only one negatively charged residue (Asp17) . This negative charge and its orientation in space were crucial for activity, since the Asp to Asn mutation and (especially) the conservative Asp to Glu mutation were deleterious . Mutations that made the peptide less cationic had, overall, less effect on the potency toward the Carnobacterium piscicola strain than on the potency toward the three other strains tested, whereas the opposite was the case for mutations that made the peptide more cationic . Thus, charged residues in the N-terminal half may - apparently via the initial electrostatic binding of the bacteriocin to target cells - influence the target-cell specificity. J Leukoc Biol, 2002 Jul, 72(1), 140 - 6 Human monocyte scavenger receptors are pattern recognition receptors for (1-->3)-beta-D-glucans; Rice PJ et al.; Glucans are cell wall constituents of fungi and bacteria that bind to pattern recognition receptors and modulate innate immunity, in part, by macrophage activation . We used surface plasmon resonance to examine the binding of glucans, differing in fine structure and charge density, to scavenger receptors on membranes isolated from human monocyte U937 cells . Experiments were performed at 25 degrees C using a biosensor surface with immobilized acetylated low density lipoprotein (AcLDL) . Inhibition of the binding by polyinosinic acid, but not polycytidylic acid, confirmed the interaction of scavenger receptors . Competition studies showed that there are at least two AcLDL binding sites on human U937 cells . Glucan phosphate interacts with all sites, and the CM-glucans and laminarin interact with a subset of sites . Polymer charge has a dramatic effect on the affinity of glucans with macrophage scavenger receptors . However, it is also clear that human monocyte scavenger receptors recognize the basic glucan structure independent of charge. Mol Cell Biol, 2002 Aug, 22(15), 5492 - 505 Nrf2 transcription factor, a novel target of keratinocyte growth factor action which regulates gene expression and inflammation in the healing skin wound; Braun S et al.; Keratinocyte growth factor (KGF) is a potent mitogen for epithelial cells, and it promotes survival of these cells under stress conditions . In a search for KGF-regulated genes in keratinocytes, we identified the gene encoding the transcription factor NF-E2-related factor 2 (Nrf2) . Nrf2 is a key player in the cellular stress response . This might be of particular importance during wound healing, where large amounts of reactive oxygen species are produced as a defense against invading bacteria . Therefore, we studied the wound repair process in Nrf2 knockout mice . Interestingly, the expression of various key players involved in wound healing was significantly reduced in early wounds of the Nrf2 knockout animals, and the late phase of repair was characterized by prolonged inflammation . However, these differences in gene expression were not reflected by obvious histological abnormalities . The normal healing rate appears to be at least partially due to an up-regulation of the related transcription factor Nrf3, which was also identified as a target of KGF and which was coexpressed with Nrf2 in the healing skin wound . T