Acta Crystallogr D Biol Crystallogr, 2001 May, 57(Pt 5), 670 - 8 Epub 2001 Apr 24. Structure of cytochrome c7 from Desulfuromonas acetoxidans at 1.9 A resolution; Czjzek M et al.; Multihaem cytochromes play a key role in electron-transport reactions in the periplasm of sulfate- and sulfur-reducing bacteria . The redox proteins grouped in the c3 superfamily also display metal-reducing activities, which make them interesting biotechnological tools . The crystal structure of the fully oxidized cytochrome c7 from Desulfuromonas acetoxidans has been solved by combined molecular-replacement and MAD methods . The structure has been refined at 1.9 A resolution to an R value of 19.1% (R(free) = 24.3%) and includes three haems and 116 water molecules . The protein displays the cytochrome c3 fold in a highly minimized form, while haem 2 and the surrounding protein environment are missing . The geometry of haem packing and of the haem axial ligands and propionates are described and compared with that of c3 cytochromes . The crystal structure is compared with the solution structure recently obtained by NMR methods and with its homologue cytochromes of the c3 superfamily . Comparison of the high number of available structures makes it possible to analyze the structural role of the few highly conserved residues, in addition to the cysteines and histidines that link the porphyrin rings and the Fe atoms to the protein chain. J Surg Res, 2001 May 1, 97(1), 65 - 70 Epithelial permeability is not increased in rats following small bowel resection; O'Brien DP et al.; BACKGROUND: Increased intestinal permeability and translocation of bacteria and/or bacterial products may cause infection and liver dysfunction in patients with the short bowel syndrome . In previous studies, serum from mice undergoing small bowel resection (SBR) enhanced growth of cultured rat intestinal epithelial cells (RIEC-6), implicating a role for a serum factor(s) in the enterocyte response to SBR . These experiments tested the hypothesis that epithelial cell permeability is increased following SBR . MATERIALS AND METHODS: Male Sprague-Dawley rats underwent a 75% SBR or sham operation . Intestinal permeability in the remnant ileum was determined by Ussing chambers on Postoperative Day (POD) 3 . Additionally, serum was collected on POD 1, 3, and 7 and mesenteric lymph was harvested on POD 3 . Once confluent, RIEC-6 cells were incubated for 3 days in media supplemented with 10% fetal bovine serum (FBS; control), 1% FBS, 1% FBS plus 9% Sham serum, or 1% FBS plus 9% SBR serum or exposed to media with varied concentrations of SBR or Sham lymph . Monolayer permeability was determined by measuring the passage of dextran-rhodamine . RESULTS: Intestinal permeability was reduced in rats undergoing SBR . Sham serum-treated monolayers demonstrated the greatest permeability . Incubation with SBR serum reduced permeability to near control media . There were no permeability differences between SBR and Sham lymph-treated monolayers . CONCLUSION: The early adaptive response of the remnant intestine after SBR is associated with reduced permeability . These results suggest an alternative mechanism for the increased bacterial translocation that has been described following SBR . Appl Environ Microbiol, 2001 May, 67(5), 2380 - 3 The celA gene, encoding a glycosyl hydrolase family 3 beta-glucosidase in Azospirillum irakense, is required for optimal growth on cellobiosides; Faure D et al.; The CelA beta-glucosidase of Azospirillum irakense, belonging to glycosyl hydrolase family 3 (GHF3), preferentially hydrolyzes cellobiose and releases glucose units from the C(3), C(4), and C(5) oligosaccharides . The growth of a DeltacelA mutant on these cellobiosides was affected . In A . irakense, the GHF3 beta-glucosidases appear to be functional alternatives for the GHF1 beta-glucosidases in the assimilation of beta-glucosides by other bacteria. Appl Environ Microbiol, 2001 May, 67(5), 2076 - 82 Case of localized recombination in 23S rRNA genes from divergent bradyrhizobium lineages associated with neotropical legumes; Parker MA; Enzyme electrophoresis and rRNA sequencing were used to analyze relationships of Bradyrhizobium sp . nodule bacteria from four papilionoid legumes (Clitoria javitensis, Erythrina costaricensis, Rhynchosia pyramidalis, and Desmodium axillare) growing on Barro Colorado Island (BCI), Panama . Bacteria with identical multilocus allele profiles were commonly found in association with two or more legume genera . Among the 16 multilocus genotypes (electrophoretic types {ETs}) detected, six ETs formed a closely related cluster that included isolates from all four legume taxa . Bacteria from two other BCI legumes (Platypodium and Machaerium) sampled in a previous study were also identical to certain ETs in this group . Isolates from different legume genera that had the same ET had identical nucleotide sequences for both a 5' portion of the 23S rRNA and the nearly full-length 16S rRNA genes . These results suggest that Bradyrhizobium genotypes with low host specificity may be prevalent in this tropical forest . Parsimony analysis of 16S rRNA sequence variation indicated that most isolates were related to Bradyrhizobium japonicum USDA 110, although one ET sampled from C . javitensis had a 16S rRNA gene highly similar to that of Bradyrhizobium elkanii USDA 76 . However, this isolate displayed a mosaic structure within the 5' 23S rRNA region: one 84-bp segment was identical to that of BCI isolate Pe1-3 (a close relative of B . japonicum USDA 110, based on 16S rRNA data), while an adjacent 288-bp segment matched that of B . elkanii USDA 76 . This mosaic structure is one of the first observations suggesting recombination in nature between Bradyrhizobium isolates related to B . japonicum versus B . elkanii. J Adhes Dent, 2000 Spring, 2(1), 9 - 28 Bonding of a self-etching primer to non-carious cervical sclerotic dentin: interfacial ultrastructure and microtensile bond strength evaluation; Tay FR et al.; PURPOSE: The objectives of this study were 1) to examine the ultrastructural features of the resin-sclerotic dentin interface following the application of Clearfil Liner Bond II sigma to natural cervical wedge-shaped lesions, and 2) to evaluate the regional tensile bond strength of this self-etching primer at different locations on natural and artificially-created cervical lesions . MATERIALS AND METHODS: Deep cervical natural lesions were bonded using the self-etching primer . Micromorphology of the bonded interface at different locations within the lesions were examined using scanning electron microscopy (SEM), transmission electron microscopy (TEM) and scanning transmission electron microscopy/energy dispersive x-ray analysis (STEM/EDX) . Ultrastructural features were further compared with the use of the same self-etching primer on artificial lesions created in sound cervical dentin . A nontrimming technique was used to evaluate the regional tensile bond strength from the occlusal, gingival, and the deepest central part of both natural and artificial cervical lesions . Beams with a mean area of 0.46 +/- 0.03 mm2 were prepared and were pulled to failure using a Bencor Multi-T testing device attached to an Instron universal tester . Bond strength results were evaluated using a two-way ANOVA design . RESULTS: A hypermineralized layer devoid of intact, banded collagen was invariably present on the surface of the natural lesions . Depending upon its thickness at different locations of the lesion, the action of a self-etching primer may be limited to this surface layer alone, producing a hybridized hypermineralized surface layer . Penetration of the self-etching primer into the underlying sclerotic dentin produced a hybridized complex containing a hybridized hypermineralized surface layer as well as a subsurface layer of hybridized intertubular dentin . Bacterial colonization of the lesion surface resulted in the formation of an additional zone of hybridized intermicrobial matrix over the surface of the lesions . Dentinal tubules remained blocked with sclerotic casts, and resin tags were rarely observed . Regional tensile bond strength results showed that the overall bond strength to natural sclerotic dentin was about 20% lower than sound cervical dentin, but was independent of the different locations within the lesions from which bond strength was evaluated . CONCLUSION: There were four factors that may have influenced the overall decrease in bond strength in natural cervical sclerotic lesions: a) the presence of a hybridized intermicrobial matrix together with entrapped bacteria may have weakened the bonds, b) inability of a self-etching primer to etch through a thick, hypermineralized surface layer, c) presence of a layer of possibly remineralized, denatured collagen at the base of the hypermineralized surface layer, and d) retention of acid-resistant sclerotic casts that obliterate the tubular lumina and prevent effective resin tag formation. Environ Mol Mutagen, 2001, 37(3), 241 - 83 Human DNA repair genes; Ronen A et al.; DNA repair systems are essential for the maintenance of genome integrity . Consequently, the disregulation of repair genes can be expected to be associated with significant, detrimental health effects, which can include an increased prevalence of birth defects, an enhancement of cancer risk, and an accelerated rate of aging . Although original insights into DNA repair and the genes responsible were largely derived from studies in bacteria and yeast, well over 125 genes directly involved in DNA repair have now been identified in humans, and their cDNA sequence established . These genes function in a diverse set of pathways that involve the recognition and removal of DNA lesions, tolerance to DNA damage, and protection from errors of incorporation made during DNA replication or DNA repair . Additional genes indirectly affect DNA repair, by regulating the cell cycle, ostensibly to provide an opportunity for repair or to direct the cell to apoptosis . For about 70 of the DNA repair genes listed in Table I, both the genomic DNA sequence and the cDNA sequence and chromosomal location have been elucidated . In 45 cases single-nucleotide polymorphisms have been identified and, in some cases, genetic variants have been associated with specific disorders . With the accelerating rate of gene discovery, the number of identified DNA repair genes and sequence variants is quickly rising . This report tabulates the current status of what is known about these genes . The report is limited to genes whose function is directly related to DNA repair . Am J Gastroenterol, 2001 Apr, 96(4), 1200 - 4 Obesity and female gender increase breath ethanol concentration: potential implications for the pathogenesis of nonalcoholic steatohepatitis; Nair S et al.; OBJECTIVES: Similarities between histological features of alcoholic hepatitis and obesity-related liver disease suggest a common pathogenic mechanism . Because intestinal bacteria can produce ethanol, it is conceivable that intestinally derived alcohol may contribute to fatty liver disease . An indirect way of measuring endogenous ethanol is to measure the breath ethanol concentration . In a previous study in ob/ob mice, breath ethanol decreased with a course of non-absorbable antibiotics, suggesting that the ethanol is derived from intestinal bacterial flora . The aims of this study were 1) to determine whether alcohol can be detected in the breath of human subjects, and 2) to assess whether there is any correlation between ethanol and obesity in patients with nonalcoholic steatohepatits (NASH) and control subjects without known liver disease . METHODS: Breath ethanol concentration was determined in 21 patients with biopsy-proven NASH and in 10 control subjects by gas chromatography . An abnormal breath ethanol level was defined as two standard deviations above the mean value of the breath ethanol of lean controls . RESULTS: Minute quantities of ethanol were detected in the breath of human subjects who had not consumed alcohol in the recent past . Patients who were obese were more likely to have higher breath ethanol concentrations . Women also had higher breath alcohol than men . However, there was no difference between patients with NASH and controls . Severity of liver disease, as evidenced by cirrhosis, did not influence the breath ethanol concentration . CONCLUSIONS: Higher breath ethanol concentrations are observed in obese subjects than in leaner ones . It is possible that intestinally derived ethanol may contribute to the pathogenesis of NASH. J Am Dent Assoc, 2001 Apr, 132(4), 482 - 91 Restorative pulpal and repair responses; Murray PE et al.; BACKGROUND: Each year, about 90 million new restorations are placed in the United States and 200 million are replaced . Controversy surrounds the pulpal reactions and frequency of bacterial microleakage associated with common restorative materials . The authors investigated and compared pulpal reactions to different types of restorative materials . METHODS: Two hundred seventy-two teeth with standardized rectangular Class V unexposed cavities were restored with resin-based composite bonded to dentin; resin-based composite bonded to enamel; resin-modified glass ionomers, or RMGI; amalgam lined with zinc polycarboxylate, or ZnPC; amalgam lined with calcium hydroxide, or Ca(OH)2; or zinc oxide-eugenol, or ZnOE . Teeth were extracted for orthodontic reasons between 20 and 381 days later . The authors categorized pulpal responses according to standards set by the Federation Dentaire Internationale and the International Organization for Standardization . Bacteria were detected using Brown-Brenn-stained sections . Pulpal responses were evaluated using histomorphometric analysis and analysis of variance statistics . RESULTS: The results showed that RMGI was the best material for preventing bacterial microleakage, and resin-based composite bonded to enamel was the worst . In regard to minimizing pulpal inflammatory activity, ZnOE was the best material and resin-based composite bonded to enamel was the worst . In terms of maximizing odontoblast survival beneath deep cavity preparations, Ca(OH)2, was the best material and RMGI was the worst . CONCLUSIONS: The results show that bacterial microleakage, pulpal injury and repair responses varied widely with different restorative materials . CLINICAL IMPLICATIONS: The authors recommend that RMGI be used to restore teeth with cavities that are shallow to moderate in depth, with the floor of deep cavities being lined with Ca(OH)2 before the teeth are restored with RMGI. Chin J Dent Res, 2000 Aug, 3(2), 44 - 8 A study of oral health condition in individuals with no oral hygiene and its association with plaque acidogenesis; Gao XJ et al.; OBJECTIVE: To study the association of long-term deposited plaque, due to lack of oral hygiene, with acidogenesis of the plaque bacteria . METHODS: Seventy-seven subjects with poor oral hygiene were selected . Debris index (DI) and calculus index (CI) were recorded . Among them, 16 were DMFS > 8, and comprised the caries active (CA) group; 27 were caries free, and comprised the caries free (CF) group . Plaque fluids in both groups were analyzed for organic acids, phosphate, and inorganic cations by use of capillary electrophoresis, while pH was measured by microelectrodes . RESULTS: No differences were found on debris index (CF group measured 2.07-0.47, CA group measured 2.01-0.53) or calculus index (CF group measured 2.47-0.50, CA group measured 2.48-0.53) relative to carious status, although there was a positive relationship between DI and CI (r = 0.52, P < 0.001) . The main finding in this study was that the quantity of lactic acid produced by sucrose exposure in these individuals with poor oral hygiene was much less (increased no more than 2 times, compared with content at rest) than in a previous report (increased 3 to 5 times, compared with content at rest) on subjects with good oral hygiene habits . CONCLUSIONS: Long-term deposited plaque due to lack of oral hygiene may have less cariogenic capability, although patients' susceptibility to periodontal disease would increase. Biotechniques, 2001 Apr, 30(4), 804 - 8, 810, 812 passim Measurement of mitochondrial pH in situ; Takahashi A et al.; In this article, we describe the advantages and disadvantages of procedures for monitoring mitochondrial pH in situ using optical microscopic techniques . The first method employs the combination of the fluorescent pH-sensitive indicator carboxy-SNARF and laser scanning confocal microscopy . Manipulation of the loading and post-loading conditions enables relatively specific accumulation of carboxy-SNARF into mitochondria . With the use of a mitochondrial-specific marker, mitochondrial pH can be accurately monitored . More recently, mitochondrial-targeted, pH-sensitive probes have been used to monitor mitochondrial pH . In particular, mitochondrial targeting of the yellow fluorescent protein (YFP) mutant of green fluorescent protein (GFP) combines the advantages of specific mitochondrial localization, high-fluorophore quantum yield, and extinction coefficient with an appropriate pKa for measuring mitochondrial pH . The use of dual-excitation ratiometry with mitochondrially targeted YFP increases the dynamic range of mitochondrial pH measurements and corrects for differences in the amount of expression of mitochondrially targeted YFP at the level of individual mitochondria. Oncogene, 2001 Jan 25, 20(4), 538 - 41 Mouse mismatch repair gene Msh2 is not essential for transcription-coupled repair of UV-induced cyclobutane pyrimidine dimers; Sonneveld E et al.; The human mutS homolog gene MSH2 is essential for DNA mismatch repair (MMR) and defects in this gene can result in increased mutagenesis, genomic instability and hereditary nonpolyposis colorectal cancer (HNPCC) . Besides correcting mismatch errors arising from DNA replication, it was shown that deficiencies in bacterial and human MMR genes including MSH2 resulted in defective transcription-coupled repair (TCR) of UV-induced photolesions . Here we show that MMR-deficient fibroblasts derived from two independent isogenic mouse strains with defined Msh2 deficiencies are as proficient in TCR of UV-induced cyclobutane pyrimidine dimers (CPD) as wildtype fibroblasts . Our results indicate that in mouse cells Msh2 is not essential for TCR of UV-induced CPD in contrast to bacteria and human cells and suggest that the biological effects of UV in mouse Msh2(-/-) cells and mice are not due to defective TCR. Cell Death Differ, 2001 Jan, 8(1), 38 - 43 Measurement of caspase activity in individual cells reveals differences in the kinetics of caspase activation between cells; Morgan MJ et al.; Most forms of apoptosis involve activation of caspases and it is likely that differences between cells in their ability to activate caspases contributes to the responsiveness of any given cell within a population to apoptotic stimuli . To study the molecular mechanisms that underlie such differences, it is necessary to measure caspase activity in individual cells . Here, we describe a method that allows the continuous monitoring of caspase activity in individual, living mammalian cells . This approach allows studies of the kinetics of caspase activation to be performed in individual cells within a population . We demonstrate that in a group of cells where some cells die and some cells survive in response to the same stimulus, the cells that die can be differentiated from those that survive based on the amount of caspase activity in each cell several hours before death occurs. Rev Hosp Clin Fac Med Sao Paulo, 2000 Nov-Dec, 55(6), 201 - 6 Ursodeoxycholic acid does not interfere with in vivo Helicobacter pylori colonization; Silva JG et al.; A low frequency of Helicobacter pylori in the gastric mucosa of patients with alkaline gastritis has been reported . At the same time, it can be noted that the growth of bacteria can be inhibited by bile acids . We studied 40 patients with chronic gastritis related to Helicobacter pylori in order to determine the effect of ursodeoxycholic acid on this infection . Diagnoses of the infection and the inflammatory process were obtained by histologic study of gastric biopsies collected during endoscopy . Two groups were studied: group I received ursodeoxycholic acid - 300 mg/day, and group II received the placebo, twice a day, both for 28 days . The colonization by Helicobacter pylori and the intensity of the mononuclear and polymorphonuclear inflammatory infiltrate were determined before (time 1) and after (time 2) treatment . Ursodeoxycholic acid had no effect on the Helicobacter pylori infection . A significant reduction in the intensity of the mononuclear inflammatory infiltrate of the gastric antrum mucosa was observed in patients from group I, when we compared not only times 1 and 2 but also groups I and II . However, this was not the case with the body mucosa . We concluded that ursodeoxycholic acid had no action on the colonization by Helicobacter pylori or on the polymorphonuclear inflammatory infiltrate, but it caused a significant reduction in the intensity of the mononuclear inflammatory infiltrate of the gastric antrum. J Biol Chem, 2001 Jun 29, 276(26), 24108 - 12 Epub 2001 Apr 19. Different mechanisms of the binding of soluble electron donors to the photosynthetic reaction center of Rubrivivax gelatinosus and Blastochloris viridis; Osyczka A et al.; The tetraheme cytochrome subunits of the photosynthetic reaction centers (RCs) in two species of purple bacteria, Rubrivivax gelatinosus and Blastochloris (Rhodopseudomonas) viridis, were compared in terms of their capabilities to bind different electron-donor proteins . The wild-type RCs from both species and mutated forms of R . gelatinosus RCs (with amino acid substitutions introduced to the binding domain for electron-donor proteins) were tested for their reactivity with soluble cytochromes and high potential iron-sulfur protein . Cytochromes from both species were good electron donors to the B . viridis RC and the R . gelatinosus RC . The reactivity in the R . gelatinosus RC showed a clear dependence on the polarity of the charges introduced to the binding domain, indicating the importance of the electrostatic interactions . In contrast, high potential iron-sulfur protein, presumed to operate according to the hydrophobic mechanism of binding, reacted significantly only with the R . gelatinosus RC . Evolutionary substitution of amino acids in a region of the binding domain on the cytochrome subunit surface probably caused the change in the principal mode of protein-protein interactions in the electron-transfer chains. FEMS Microbiol Lett, 2001 Apr 13, 197(2), 215 - 21 TRAM, a predicted RNA-binding domain, common to tRNA uracil methylation and adenine thiolation enzymes; Anantharaman V et al.; A previously undetected conserved domain is identified in two distinct classes of tRNA-modifying enzymes, namely uridine methylases of the TRM2 family and enzymes of the MiaB family that are involved in 2-methylthioadenine formation . This domain, for which the acronym TRAM is proposed after TRM2 and MiaB, is predicted to bind tRNA and deliver the RNA-modifying enzymatic domains to their targets . In addition to the two families of RNA-modifying enzymes, the TRAM domain is present in several other proteins associated with the translation machinery and in a family of small, uncharacterized archaeal proteins that are predicted to have a role in the regulation of tRNA modification or translation . Secondary structure prediction indicates that the TRAM domain adopts a simple beta-barrel fold . In addition, sequence analysis of the MiaB family enzymes showed that they share the predicted catalytic site with biotin and lipoate synthases and probably employ the same mechanism for sulfur insertion into their respective substrate. J Agric Food Chem, 2001 Mar, 49(3), 1639 - 47 Metabolism and distribution of {2,3-(14)C}acrolein in laying hens; Sharp DE et al.; The metabolism and distribution of {2,3-(14)C}-acrolein were studied in 10 laying hens orally administered 1.09 mg/kg of body weight/day for 5 days . Eggs, excreta, and expired air were collected . The hens were killed 12-14 h after the last dose and edible tissues collected . The nature of radioactive residues was determined in tissues and eggs . All of the identified metabolites were the result of the incorporation of acrolein-derived radioactivity into normal natural products of intermediary metabolism in the hen except for 1,3-propanediol, which is a known degradation product of glycerol in bacteria. Bioconjug Chem, 2001 Mar-Apr, 12(2), 152 - 62 Biotinylated indoles as probes for indole-binding proteins; Dolusic E et al.; Biotinylated indoles were prepared for application as bifunctional probes for the detection of indole-binding proteins which participate in the life processes of humans, animals, plants, and bacteria . The indole nucleus was functionalized, at ring positions 3, 5, or 6, by attachment of a 2-aminoethyl group, which was then coupled to the carboxyl moiety of biotin, via a spacer composed of 3 or 4 concatenated beta-alanine residues . The constructs thus obtained were able to inhibit tryptophanase activity, similarly to indole in a concentration-dependent manner . They also bound strongly to lysozyme and weakly to bovine and human serum albumins, in accordance with the known affinities of these proteins for indole and 3-(2-aminoethyl)indole (tryptamine) . The biotin end of the protein-bound bifunctional probes could then be detected by coupling to (strept)avidin conjugated to alkaline phosphatase or horseradish peroxidase, followed by incubation with substrates which are converted by these enzymes to intensely colored or chemiluminescent products. J Theor Biol, 2001 Apr 7, 209(3), 345 - 9 The non-universality of the genetic code: the universal ancestor was a progenote; Di Giulio M; The coevolution theory of genetic code origin (Wong, J.T . 1975, Proc . Natl Acad . Sci . U.S.A.72, 1909-1912) is assumed here to be substantially correct . This theory is based on the strict parallelism of the biosynthetic relationships between amino acids and the organization of the genetic code and postulates that these relationships were mediated by tRNA-like molecules on which the biosynthetic transformations between precursor and product amino acids took place . These transformations underlay the mechanism that gave rise to genetic code organization . One of the pathways which represents these transformations found in current organisms, and which are thus probably molecular fossils, is the Met-tRNA(fMet)-->fMet-tRNA(fMet)pathway . This pathway is present only in the Bacteria domain . This along with other observations and arguments leads us to believe that this pathway is a clear violation of the universality of the genetic code . Furthermore, the presence of this pathway only in the Bacteria domain seems to imply that the translation apparatus was still rapidly evolving when this pathway was fixed . This, in turn, appears to imply that the last universal common ancestor was a progenote . Finally, the implications that the finding of this pathway has for the stereochemical theory of genetic code origin are discussed . Cytokine Growth Factor Rev, 2001 Mar, 12(1), 53 - 72 Interleukin-18 is a unique cytokine that stimulates both Th1 and Th2 responses depending on its cytokine milieu; Nakanishi K et al.; IL-18 is a potent proinflammatory cytokine able to induce IFNgamma, GM-CSF, TNFalpha and IL-1 in immunocompetent cells, to activate killing by lymphocytes, and to up-regulate the expression of certain chemokine receptors . IL-18 is also essential to host defences against severe infections . In particular, the clearance of intracellular bacteria, fungi and protozoa requires the induction of host-derived IFNgamma, which evokes effector molecules such as nitric oxide . Also, IL-18 plays a part in the clearance of viruses, partly by the induction of cytotoxic T cells, and the expulsion of viruses is impaired in IL-18-deficient mice . IL-18 also enhances tumour rejection by its potent capacity to augment the cytotoxic activity of NK and T cells in vivo . In contrast, recent studies also demonstrate a convincing role for IL-18 in atopic responses, including atopic asthma . IL-18 induces naive T cells to develop into Th2 cells . Moreover, IL-18 also induces IL-13 and/or IL-4 production by NK cells, mast cells and basophils . Therefore, IL-18 should be seen as a unique cytokine that enhances innate immunity and both Th1- and Th2-driven immune responses. Peptides, 2001 Apr, 22(4), 575 - 82 Functional analysis of glucose-dependent insulinotropic polypeptide fusion proteins; Ding KH et al.; To generate functional fluorescently tagged glucose-dependent insulinotropic polypeptide (GIP), a series of GIP expression constructs were devised . These included G1 (complete preprohormone), G2 (lacking the C-terminal extension), G3 (lacking both N- and C-terminal extensions), G4 (G2 fused to green fluorescent protein, GFP), and G5 (G3 fused to GFP) . Expression of G5 in bacteria generated immunopositive GIP together with GFP fluorescence, while G4 generated only fluorescence without immunoreactivity . Transfection of NIH3T3 cells with cDNAs of G1, G3, G5, but not G2, G4, and EGFP, resulted in immunologically detectable GIP formation, although fluorescence could be detected in the latter two . GIP as well as GIP-GFP secreted by NIH3T3 cells significantly stimulated intracellular cAMP accumulation and Ca(2+) mobilization in SaOS2 cells . The GIP receptor antagonist GIP(7-30) abolished these responses . These results suggest that a GIP-GFP fusion protein seven times larger than the native peptide retains function and may be used as an in vivo probe to detect GIP receptor distribution and to explore GIP's biological roles. Gene, 2001 Apr 4, 267(1), 135 - 44 Okadaic acid induces transcription of junB through a CCAAT box and NF-Y; Finch JS et al.; The shellfish toxin, okadaic acid (OA), is a potent tumor promoter that induces expression of the proto-oncogene junB in mouse keratinocyte 308 cells . Here we show, through deletion analysis of the junB promoter, that sequences near the TATA box conferred transcriptional induction by OA . Transient transfections of luciferase constructs bearing the junB promoter with single mutations in various cis elements demonstrated that a promoter containing a mutated CCAAT box could not be induced by OA . When this CCAAT box was inserted into a heterologous promoter construct, OA induction was dependent on an intact CCAAT box . Flanking cis elements located near the CCAAT box, although not required for OA inducibility, did play a role in the basal level of transcription . NF-Y was shown by EMSA to bind to the CCAAT box . OA induction from the junB CCAAT box was blocked by dominant negative NF-YA as well as the CCAAT box-dependent anticancer drug, ET-473 . Expression of a lexA/NF-YA chimeric protein demonstrated that OA induction was dependent on the binding of NF-Y family members . These studies demonstrate that OA can mediate transcriptional activation of junB through the classical CCAAT box and that transcription factor NF-Y plays a functional role in the induction. FEMS Microbiol Ecol, 2001 May, 35(3), 231 - 238 Inhibition of methanogens increases photo-dependent nitrogenase activities in anoxic paddy soil amended with rice straw; Harada N et al.; The interaction between phototrophic dinitrogen fixers and methanogens was examined in soil slurries amended with rice straw using 2-bromoethanesulfonic acid (BES), a specific methanogenic inhibitor . Slurries incubated in light increased phototrophic nitrogenase activity (acetylene reducing activity), and showed growth of phototrophic purple bacteria and reduction of CH(4) emission, indicating outcompetition of purple bacteria with methanogens in photic zones . Adding BES effectively inhibited methane production and markedly increased phototrophic acetylene reducing activity accompanied with acetate accumulation, but did not affect populations of purple bacteria in the slurries . More acetate accumulated in the inhibited slurries incubated in dark . We suggest that increased availability of organic substrates for purple bacteria after stopping methanogenic consumption by BES caused the increased phototrophic acetylene reducing activity . These results indicate that, after purple bacteria grow enough, performance of their N(2) fixation may be limited by substrate availability, which methanogenesis may profoundly influence. Rev Neurol, 2001 Feb 1-15, 32(3), 201 - 6 {Chlamydia pneumoniae and cerebrovascular disease}; Linares-Palomino J et al.; INTRODUCTION: Chlamydia pneumoniae has relationship with artherosclerosis of carotid artery by seroepidemiological studies and by demonstration of the bacteria in atheromata . We made a case-control study to know the seroprevalence of chronic infection of C . pneumoniae and the presence of the bacteria in arterial biopsies . PATIENTS AND METHODS: The cases group was constituted by 26 patients undergoing carotid surgery . In the control group there were 50 patients without atherosclerosis and who underwent removal of their varicose veins . There were matched for sex, age and smoking . We obtained serum samples to determinate IgG antibodies against MOMP by MIF and ELISA . In the cases group, we got the arterial biopsies from carotid artery, and from pudendal arteries in control group . We determined chlamydial DNA on the biopsies by heminested PCR (primers: HL-1, HM-1, HR-1) . We measured fibrinogen in both groups . RESULTS: By MIF technique, the seroprevalence was (IgG > 1:32) 69.23% cases versus 24% controls (OR: 7.12, CI: 2.47-20.48) . The ELISA showed 76.92% of seropositivity in cases versus 16% in controls (OR: 17.5, CI: 95%: 5.35-57.23) . The DNA of C . pneumoniae was found in 18 cases and 6 controls, (p < 0.0001, chi 2) . We did not find any relationship between fibrinogen levels and groups . CONCLUSION: We think that there is a relationship between chronic infection with C . pneumoniae and carotid atherosclerosis. Mol Microbiol, 2001 Apr, 40(2), 451 - 64 MspA provides the main hydrophilic pathway through the cell wall of Mycobacterium smegmatis; Stahl C et al.; MspA is an extremely stable, oligomeric porin from Mycobacterium smegmatis that forms water-filled channels in vitro . Immunogold electron microscopy and an enzyme-linked immunosorbent assay demonstrated that MspA is localized in the cell wall . An mspA deletion mutant did not synthesize detectable amounts of mspA mRNA, as revealed by amplification using mspA-specific primers and reverse-transcribed RNA . Detergent extracts of the DeltamspA mutant exhibited a significantly lower porin activity in lipid bilayer experiments and contained about fourfold less porin than extracts of wild-type M . smegmatis . The chromosome of M . smegmatis encodes three proteins very similar to MspA . Sequence analysis of the purified porin revealed that mspB or mspC or both genes are expressed in the DeltamspA mutant . The properties of this porin, such as single channel conductance, extreme stability against denaturation, molecular mass and composition of 20 kDa subunits, are identical to those of MspA . Deletion of mspA reduced the cell wall permeability towards cephaloridine and glucose nine- and fourfold respectively . These results show that MspA is the main general diffusion pathway for hydrophilic molecules in M . smegmatis and was only partially replaced by fewer porins in the cell wall of the DeltamspA mutant . The minimal permeability coefficient of the DeltamspA mutant for glucose was 7.2 x 10(-8) cm s(-1), which is the lowest value reported so far for bacteria . This is the first experimental evidence that porins are the major determinants of the exceptionally low permeability of mycobacteria to hydrophilic molecules. Rinsho Byori, 2001 Feb, 49(2), 109 - 15 {The history of Helicobacter pylori}; Fukuda Y et al.; Histological and ultrastructural studies of gastric mucosa with spiral bacteria had been published at the Royal Perth Hospital of Western Australia in 1979 . The pathologist Warren correlated them with inflammation . In 1981, Marshall was training in internal medicine . Warren, Marshall and Goodwin started culture of bacteria, but spiral bacteria were not cultured . The 35th culture was left during the Easter holiday, and after 5 days 1-mm transparent colonies were seen on the plate . Since discovery Helicobacter pylori(H . pylori) have continued to fascinate and challenge doctors and scientists for 18 years to come . In 2000, triple therapy with PPI, Amoxicillin and clarithromycin was approved for treatment of peptic ulcer disease in Japan. Int Endod J, 2000 Jul, 33(4), 320 - 5 Cleaning efficacy of a new root canal irrigation solution: a preliminary evaluation; Marais JT; AIM: A new product, electro-chemically activated water, was compared to NaOCl for its cleaning effect on root canal walls . METHODOLOGY: Root canal treatment was carried out on two groups of extracted teeth with one of the irrigants being used in each group . The control group received no treatment . All teeth were split and the canal walls viewed in a scanning electron microscope . RESULTS: The canal walls of the control group were covered by debris and bacteria . Sodium hypochlorite produced clean surfaces with the dentinal tubules open in some areas and occluded by the smear layer in other areas; in some areas bacteria were visible inside or under the smear layer . Electro-chemically activated water produced markedly cleaner surfaces, removing the smear layer in large areas . CONCLUSIONS: The cleaning efficacy of electro-chemically activated water in root canals was considered to be superior to NaOCl. Chem Biol Interact, 2001 Jan 30, 130-132(1-3), 219 - 25 Inhibition of cytosolic class 3 aldehyde dehydrogenase by antisense oligonucleotides in rat hepatoma cells; Muzio G et al.; Aldehyde dehydrogenases (ALDHs) are a superfamily of several isoenzymes widely expressed in bacteria, yeast, plant and animals . Three major classes of ALDHs have been traditionally identified, classes 1, 2 and 3 . Both exogenous and endogenous aldehydes, including aldehydes derived from lipid peroxidation, are oxidized by the ALDH superfamily . Several changes in ALDH isoenzyme expression take place in hepatoma cells, in particular cytosolic class 3 ALDH (ALDH3), not expressed in normal hepatocytes, appears and increases with the degree of deviation . It has been demonstrated that cytosolic ALDH3 is important in determining the resistance of tumor cells to antitumor drugs, such as cyclophosphamide . Moreover, hepatoma-associated ALDH3 seems to be important in metabolizing aldehydes derived from lipid peroxidation, and in particular the cytostatic aldehyde 4-hydroxynonenal (4-HNE) . We demonstrated previously that restoring endogenous lipid peroxidation in hepatoma cells by enriching them with arachidonic acid causes a decrease of mRNA, protein and enzyme activity of ALDH3 and that this decrease reduces cell growth and/or causes cell death, depending on basal class 3 ALDH activity . To confirm the correlation between inhibition of class 3 ALDH and reduction of cell proliferation, we exposed hepatoma cells to antisense oligonucleotides (ODNs) against ALDH3 . In JM2 hepatoma cell line, with high ALDH3 activity, the exposure to antisense ODNs significantly decreases mRNA and enzyme activity (90%) . At the same time, cell growth was reduced by about 70% . The results confirm that in hepatoma cells ALDH3 expression is closely related with cell growth, and that its inhibition is important in reducing the proliferation of hepatoma cells overexpressing ALDH3. Genome Biol . 2001;2(4):RESEARCH0010 . Epub 2001 Mar 22. A simple model based on mutation and selection explains trends in codon and amino-acid usage and GC composition within and across genomes; Knight RD et al.; BACKGROUND: Correlations between genome composition (in terms of GC content) and usage of particular codons and amino acids have been widely reported, but poorly explained . We show here that a simple model of processes acting at the nucleotide level explains codon usage across a large sample of species (311 bacteria, 28 archaea and 257 eukaryotes) . The model quantitatively predicts responses (slope and intercept of the regression line on genome GC content) of individual codons and amino acids to genome composition . RESULTS: Codons respond to genome composition on the basis of their GC content relative to their synonyms (explaining 71-87% of the variance in response among the different codons, depending on measure) . Amino-acid responses are determined by the mean GC content of their codons (explaining 71-79% of the variance) . Similar trends hold for genes within a genome . Position-dependent selection for error minimization explains why individual bases respond differently to directional mutation pressure . CONCLUSIONS: Our model suggests that GC content drives codon usage (rather than the converse) . It unifies a large body of empirical evidence concerning relationships between GC content and amino-acid or codon usage in disparate systems . The relationship between GC content and codon and amino-acid usage is ahistorical; it is replicated independently in the three domains of living organisms, reinforcing the idea that genes and genomes at mutation/selection equilibrium reproduce a unique relationship between nucleic acid and protein composition . Thus, the model may be useful in predicting amino-acid or nucleotide sequences in poorly characterized taxa. Biol Pharm Bull, 2001 Apr, 24(4), 351 - 6 Interaction of phytoestrogens with estrogen receptors alpha and beta; Morito K et al.; The human estrogen receptor (hER) exists as two subtypes, hER alpha and hER beta, that differ in the C-terminal ligand-binding domain and in the N-terminal transactivation domain . In this study, we investigated the estrogenic activities of soy isoflavones after digestion with enteric bacteria in competition binding assays with hER alpha or hER beta protein, and in a gene expression assay using a yeast system . The estrogenic activities of these isoflavones were also investigated by the growth of MCF-7 breast cancer cells . Isoflavone glycoside binds weakly to both receptors and estrogen receptor-dependent transcriptional expression is poor . The aglycones bind more strongly to hER beta than to hER alpha . The binding affinities of genistein, dihydrogenistein and equol are comparable to the binding affinity of 17 beta-estradiol . Equol induces transcription most strongly with hER alpha and hER beta . The concentration required for maximal gene expression is much higher than expected from the binding affinities of the compounds, and the maximal activity induced by these compounds is about half the activity of 17 beta-estradiol . Although genistin binds more weakly to the receptors and induces transcription less than does genistein, it stimulates the growth of MCF-7 cells more strongly than does genistein. Phys Rev E Stat Nonlin Soft Matter Phys . 2001 Jan;63(1 Pt 1):011903 . Epub 2000 Dec 20. Correlation property of length sequences based on global structure of the complete genome; Yu ZG et al.; This paper considers three kinds of length sequences of the complete genome . Detrended fluctuation analysis, spectral analysis, and the mean distance spanned within time L are used to discuss the correlation property of these sequences . The values of the exponents from these methods of these three kinds of length sequences of bacteria indicate that the long-range correlations exist in most of these sequences . The correlations have a rich variety of behaviors including the presence of anti-correlations . Furthermore, using the exponent gamma, it is found that these correlations are all linear (gamma=1.0+/-0.03) . It is also found that these sequences exhibit 1/f noise in some interval of frequency (f>1) . The length of this interval of frequency depends on the length of the sequence . The shape of the periodogram in f>1 exhibits some periodicity . The period seems to depend on the length and the complexity of the length sequence. Ann Pharmacother, 2001 Apr, 35(4), 424 - 6 Life-threatening eosinophilic pleuropericardial effusion related to vitamins B5 and H; Debourdeau PM et al.; OBJECTIVE: To report a case of eosinophilic pleuropericarditis resulting from concomitant use of vitamins B5 and H . CASE SUMMARY: A 76-year-old white woman was admitted to the hospital because of chest pain and dyspnea related to pleurisy and a pericardial tamponade . This patient had no history of allergy and had been taking vitamins B5 and H for two months . Blood tests performed showed an inflammatory syndrome and a high eosinophil concentration (1200-1500 cells/mm3) . Pleurocentesis and pericardiotomy yielded a sterile exudative fluid with an eosinophilic infiltrate . There were no nuclear antibodies and no rheumatic factor; screenings for viruses, parasites, bacteria, and malignant tumor were negative . A myelogram, biopsy of the iliac crest bone, and concentration of immunoglobulin E were also normal . After withdrawal of the vitamins, the patient recovered and the eosinophilia disappeared . DISCUSSION: Prolonged hypereosinophilia has marked predilection to damage specific organs, including the heart, but pleuropericardial effusion is uncommon . Drug-related pleuropericarditis usually occurs without an increased eosinophil count . Other drugs responsible for eosinophilic pleuropericarditis are cephalosporins, dantrolene, propylthiouracil, and nitrofurantoin . To our knowledge, this is the first case report of pleuropericarditis related to vitamins B5 and H . CONCLUSIONS: This case suggests that vitamins B5 and H may cause symptomatic, life-threatening, eosinophilic pleuropericarditis . Physicians prescribing these commonly used vitamins should be aware of this potential adverse reaction. Z Naturforsch {C}, 2001 Jan-Feb, 56(1-2), 1 - 5 Secondary metabolites from a Streptomyces strain isolated from Livingston Island, Antarctica; Ivanova V et al.; The producing strain Streptomyces sp . 1010 was isolated from a shallow sea sediment from the region of Livingston Island, Antarctica . From the culture broth of this strain naturally active secondary metabolites were isolated identical to phthalic acid diethyl ester (C12H14O4, MW . 222); 1, 3-bis (3-phenoxyphenoxy)benzene (C30H22O4, MW.446); hexanedioic acid dioctyl ester (C22H42O4, MW.370) and the new substance 2-amino- 9, 13 -dimethyl heptadecanoic acid (C19H39NO2, MW.313) . These compounds represent diverse classes of chemical structures and provide evidence for the untapped biosynthetic potential of marine bacteria from Antarctica. Biosci Biotechnol Biochem, 2001 Feb, 65(2), 254 - 63 New classification system for oxygenase components involved in ring-hydroxylating oxygenations; Nam JW et al.; Batie et al . {Chemistry and Biochemistry of Flavoenzymes, 3, 543-556 (1991)} proposed a classification system for ring-hydroxylating oxygenases in which the oxygenases are grouped into three classes in terms of the number of constituent components and the nature of the redox centers . But in recent years, many ring-hydroxylating oxygenases have been newly identified and characterized, and found difficult to classify into these three classes . Typical examples are carbazole 1,9a-dioxygenase and 2-oxo-1,2-dihydroquinoline 8-monooxygenase, which have been classified into class III and class IB, respectively, from biochemical characteristics . However, a phylogenetic study showed that the terminal oxygenases of both are closely related to class IA . Because this discrepancy derived from counting all the components together, here we proposed a new scheme based on the homology of the amino acid sequences of the alpha subunits of the terminal oxygenase components . This new scheme strongly reflects the actual phylogenetic affiliation of the terminal oxygenase component . By comparing their sequences pairwise using the CLUSTAL W program, 54 oxygenase components were classified into 4 groups (groups I, II, III, and IV) . While group I contains broad-range oxygenases sharing low homology, groups II, III, and IV contain some typical oxygenases: benzoate/toluate dioxygenases for group II, naphthalene/polycyclic aromatic hydrocarbon dioxygenases for group III, and benzene/toluene/biphenyl dioxygenases for group IV . Our new scheme is simple and powerful, since an oxygenase component can be nearly automatically grouped when the DNA sequence is available, and it fits very well with the phylogenetic affiliation. Microsc Res Tech, 2001 Apr 15, 53(2), 147 - 56 Elemental maps from EFTEM images using two different background subtraction models; Quintana C et al.; Acquisition of a great number of energy-filtered images in a TEM (EFTEM) around the characteristic signal with a low energy-selecting slit allows display of the electron energy loss (EEL)-spectrum of regions of interest (ROIs) of a sample . These EEL-spectra can be submitted to the different treatments already in use for electron energy loss spectroscopy (EELS) . In particular, it is possible to fit the experimental background with different mathematical models, using images acquired below and above a characteristic ionization edge . After this fitting, elemental maps can be computed by subtraction of the extrapolated/interpolated background from the characteristic images . In this work, we compared two mathematical models for background fitting-the Egerton power law and the log-polynomial law . We studied the low-energy region (40-150 eV) and a higher-energy region (350-600 eV) with the aid of software for interactive processing of EFTEM image series that we developed . The analyzed elements were the constitutive elements: iron, phosphorus, nitrogen, and oxygen in several biological materials . Two analytical TEMs, one equipped with a post-column and the other with an in-column spectrometer, were used . Our experimental results confirm that the power law is very sensitive to the value of the energy loss of the pre-edge images when the background is computed by extrapolation . The log-polynomial model is less sensitive than the power law model to the value of the energy loss of the pre-edge images in the low energy region . For the oxygen K edge at 535 eV, it gives the best fit when it is combined with the interpolation method . The use of programs that facilitate the handling of EFTEM image series, and the controlled calculation of the background under the characteristic images, represent a step forward in the generation of elemental maps . Curr Biol, 2001 Mar 20, 11(6), 431 - 5 Recombination in Wolbachia; Werren JH et al.; Wolbachia are widely distributed intracellular bacteria that cause a number of reproductive alterations in their eukaryotic hosts . Such alterations include the induction of parthenogenesis, feminization, cytoplasmic incompatibility, and male killing {1-11} . These important bacteria may play a role in rapid speciation in insects {12-14}, and there is growing interest in their potential uses as tools for biological control and genetic manipulation of pests and disease vectors {15-16} . Here, we show recombination in the Wolbachia outer surface protein gene (wsp) between strains of Wolbachia . In addition, we find a possible ecological context for this recombination . Evidence indicates either genetic exchange between Wolbachia in a parasitoid wasp and in the fly that it parasitizes or horizontal transfer of Wolbachia between the parasitoid and the fly, followed by a recombination event . Results have important implications for the evolution of these bacteria and the potential use of Wolbachia in biological control. BMC Dev Biol . 2001;1(1):4 . Epub 2001 Mar 27. Cre reporter strains produced by targeted insertion of EYFP and ECFP into the ROSA26 locus; Srinivas S et al.; BACKGROUND: Several Cre reporter strains of mice have been described, in which a lacZ gene is turned on in cells expressing Cre recombinase, as well as their daughter cells, following Cre-mediated excision of a loxP-flanked transcriptional "stop" sequence . These mice are useful for cell lineage tracing experiments as well as for monitoring the expression of Cre transgenes . The green fluorescent protein (GFP) and variants such as EYFP and ECFP offer an advantage over lacZ as a reporter, in that they can be easily visualized without recourse to the vital substrates required to visualize beta-gal in living tissue . RESULTS: In view of the general utility of targeting the ubiquitously expressed ROSA26 locus, we constructed a generic ROSA26 targeting vector . We then generated two reporter lines of mice by inserting EYFP or ECFP cDNAs into the ROSA26 locus, preceded by a loxP-flanked stop sequence . These strains were tested by crossing them with transgenic strains expressing Cre in a ubiquitous (beta-actin-Cre) or a cell-specific (Isl1-Cre and En1-Cre) pattern . The resulting EYFP or ECFP expression patterns indicated that the reporter strains function as faithful monitors of Cre activity . CONCLUSIONS: In contrast to existing lacZ reporter lines, where lacZ expression cannot easily be detected in living tissue, the EYFP and ECFP reporter strains are useful for monitoring the expression of Cre and tracing the lineage of these cells and their descendants in cultured embryos or organs . The non-overlapping emission spectra of EYFP and ECFP make them ideal for double labeling studies in living tissues. Mol Ecol, 2001 Mar, 10(3), 703 - 9 Do Wolbachia infections play a role in unidirectional incompatibilities in a field cricket hybrid zone? Mandel MJ, Ross CL, Harrison RG. Two closely related field crickets, Gryllus firmus and G . pennsylvanicus, hybridize along an extensive north-south zone in the eastern United States . Crosses between G . firmus males and G . pennsylvanicus females produce viable and fertile F1, but the reciprocal cross consistently fails to produce offspring . Wolbachia, a bacterial parasite of arthropods that causes unidirectional incompatibilities in a variety of insect species, has been suggested as the cause of the observed incompatibility between G . pennsylvanicus and G . firmus . We examine the presence/absence of Wolbachia strains, defined by sequencing the ftsZ gene, in four cricket populations from the north-eastern United States . Most G . firmus individuals are infected (100% in Guilford, Connecticut; 65% in Seaside Park, New Jersey) and > 95% of those infected harbour a single strain of Wolbachia . All individuals in G . pennsylvanicus populations (Ithaca, New York; Sharon, Connecticut) are infected; the majority of individuals carry a second strain of Wolbachia, but a significant fraction carry the same strain found commonly in G . firmus . The presence of an apparently identical Wolbachia strain in crickets of both species means that some crosses between G . pennsylvanicus males and G . firmus females should be compatible . We have no evidence of such compatibility . Furthermore, if Wolbachia infections are responsible for the observed incompatibility between species, then incompatibilities must also exist within G . pennsylvanicus, because this species harbours both Wolbachia strains . Although some single pair crosses within G . pennsylvanicus do fail to produce offspring, the proportion is lower than expected if Wolbachia were responsible . Therefore, Wolbachia is unlikely to be involved in reproductive isolation between the two cricket species. Eur J Immunol, 2001 Apr, 31(4), 1268 - 77 Immune protection against septic peritonitis in endotoxin-primed mice is related to reduced neutrophil apoptosis; Feterowski C et al.; The innate immune system provides essential information about the presence of infectious danger and signals the activation and instruction of adaptive immunity . The present study addressed the question of whether prior exposure of the innate immune system to LPS may modulate host defense against acute septic peritonitis . We show that LPS priming 4 days, but not 2 days, prior to infection enhances bacterial clearance and improves survival of septic peritonitis . Immune protection in day 4 LPS-primed mice was specifically associated with a marked increase in the accumulation and activation of neutrophils at the site of infection . Accumulating neutrophils in day 4 LPS-primed mice exhibited a normal production of reactive oxygen metabolites in response to in vivo exposure to intestinal bacteria . The local increase in neutrophil numbers was found to result from a reduced rate of apoptotic cell death . Inhibition of neutrophil apoptosis in LPS-primed mice was mediated by soluble factor(s) distinct from G-CSF and GM-CSF . Thus, engagement of pattern recognition systems prior to infection may improve host defense by amplifying the effector cell response of innate immunity . The results also provide in vivo evidence that apoptosis of inflammatory cells represents an important process for the control of host defense to infection. Mol Microbiol, 2001 Apr, 40(1), 225 - 34 Cell cycle regulation in the hyperthermophilic crenarchaeon Sulfolobus acidocaldarius; Hjort K et al.; The regulation and co-ordination of the cell cycle of the hyperthermophilic crenarchaeon Sulfolobus acidocaldarius was investigated with antibiotics . We provide evidence for a core regulation involving alternating rounds of chromosome replication and genome segregation . In contrast, multiple rounds of replication of the chromosome could occur in the absence of an intervening cell division event . Inhibition of the elongation stage of chromosome replication resulted in cell division arrest, indicating that pathways similar to checkpoint mechanisms in eukaryotes, and the SOS system of bacteria, also exist in archaea . Several antibiotics induced cell cycle arrest in the G2 stage . Analysis of the run-out kinetics of chromosome replication during the treatments allowed estimation of the minimal rate of replication fork movement in vivo to 250 bp s-1 . An efficient method for the production of synchronized Sulfolobus populations by transient daunomycin treatment is presented, providing opportunities for studies of cell cycle-specific events . Possible targets for the antibiotics are discussed, including topoisomerases and protein glycosylation. Clin Microbiol Infect, 2001 Feb, 7(2), 75 - 9 Survival of Coxiella burnetii within free-living amoeba Acanthamoeba castellanii; La Scola B et al.; OBJECTIVE: To determine whether Coxiella burnetti is able to survive within free-living amoeba . METHODS: When C . burnetii was co-cultivated with the free-living amoeba species Acanthamoeba castellanii . RESULTS: Viable bacteria were detected for 18 days . Microscopic studies confirmed the presence of bacteria within vacuoles, some of which appeared to be spore-like forms . CONCLUSION: These results indicate that free-living amoebae could provide an intracellular niche for the spore formation and survival of C . burnetii in the environment. J Biol Chem, 2001 Apr 27, 276(17), 14178 - 86 Epub 2001 Jan 31. Identification of the motif in versican G3 domain that plays a dominant-negative effect on astrocytoma cell proliferation through inhibiting versican secretion and binding; Wu Y et al.; This study was designed to investigate the mechanisms by which mutant versican constructs play a dominant-negative effect on astrocytoma cell proliferation . Although a mini-versican or a versican G3 construct promoted growth of U87 astrocytoma cells, a mini-versican lacking epidermal growth factor (EGF) motifs (versicanDeltaEGF) and a G3 mutant (G3DeltaEGF) exerted a dominant-negative effect on cell proliferation . G3DeltaEGF-transfected cells formed smaller colonies, arrested cell cycle at G(1) phase, inhibited expression of cell cycle proteins cdk4 and cyclin D1, and contained multiple nucleoli . In cell surface binding assays, G3 products expressed in COS-7 cells and bacteria bound to U87 cell surface . G3DeltaEGF products exhibited decreased binding activity, but higher levels of G3DeltaEGF products were able to inhibit the binding of G3 to the cell surface . G3DeltaEGF expression inhibited secretion of endogenous versican in astrocytoma cells and also inhibited the secretion of mini-versican in COS-7 cells co-transfected with the mini-versican and G3DeltaEGF constructs . The effect seems to depend on the expression efficiency of G3DeltaEGF, and it occurred via the carbohydrate recognition domain. Keio J Med, 2001 Mar, 50(1), 39 - 44 Characterization of CD1d in mucosal immune function: an immunotherapeutic target for inflammatory bowel disease; Blumberg RS; In addition to the classical MHC class I and class II molecules, human intestinal epithelial cells also express nonclassical MHC class I-like molecules on their cell surface . CD1d is a non-polymorphic MHC-like molecule whose expression is mainly localized to the epithelial cells of the gastrointestinal tract . The biochemical structure of CD1d on intestinal epithelial cells (IECs) exists in two forms: a 37-kD nonglycosylated, beta 2-microglobulin (beta 2M) independent and a 48-kD glycosylated, beta 2M dependent form . Immunolocalization studies suggest that the 37-kD nonglycosylated form of CD1d is limited to the apical cell surface whereas the 48-50-kD glycosylated, beta 2M dependent form of CD1d is expressed both on the apical and the basolateral surfaces . The beta 2M association with CD1d seems to be important in regulating the pattern of glycosylation and the localization of CD1d within the cell based upon studies of the structure of CD1d in a transfected model cell line and in polarized epithelial cell monolayers . The functional role of intestinal CD1d remains unknown . However, based upon in vitro studies of the antigens presented by human CD1d and mouse CD1d, CD1d expressed on IECs likely presents a very hydrophobic glycolipid molecule possibly from the cell wall of bacteria or host cells . The processed-lipid antigen presented by CD1d may then involve a yet-to-be-identified subpopulation of the resident, oligoclonal alpha beta TCR CD8+ intestinal intraepithelial lymphocyte (iIEL) T cells . Subsequently, these T cells would be very important in regulating the local immune response by producing cytokines and recruiting other immune modulating cells to destroy infected cells, regenerate normal IECs, and possibly downregulate activated T cells to maintain mucosal integrity. Chest, 2001 Apr, 119(4), 1283 - 5 Minocycline-induced pancreatitis in cystic fibrosis; Boyle MP; We report two cases of acute pancreatitis secondary to minocycline use in adults with cystic fibrosis (CF) . This minocycline complication has not previously been reported . Given the increased use of minocycline in the adult CF population to treat resistant bacteria, awareness of this potential adverse effect is imperative . As both of these individuals with CF had class IV genotypes and pancreatic sufficiency, close observation is warranted in the future to determine if persons with pancreatic-sufficient CF are at an increased risk for minocycline-induced pancreatitis. FEMS Microbiol Ecol, 2001 Apr, 35(2), 163 - 169 Is interspecies hydrogen transfer needed for toluene degradation under sulfate-reducing conditions? Elshahed MS, McInerney MJ. Sediments from a hydrocarbon-contaminated aquifer, where periodic shifts between sulfate reduction and methanogenesis occurred, were examined to determine whether the degradation of toluene under sulfate-reducing conditions depended on interspecies hydrogen transfer . Toluene degradation under sulfate-reducing conditions was inhibited by the addition of 5 mM sodium molybdate, but the activity was not restored upon the addition of an actively growing, hydrogen-using methanogen . Toluene degradation was not inhibited in microcosms where hydrogen levels were maintained at a level theoretically sufficient to inhibit toluene degradation if the process proceeded via interspecies hydrogen transfer . Finally, the addition of carbon monoxide, a potent inhibitor of hydrogenase activity, inhibited hydrogen but not toluene consumption in sulfate-reducing microcosms . These results suggest that toluene is degraded directly by sulfate-reducing bacteria without the involvement of interspecies hydrogen transfer . The sequence of experiments used to reach this conclusion could be applied to determine the role of interspecies hydrogen transfer in the degradation of a variety of compounds in different environments or under different terminal electron-accepting conditions. Mol Biol Cell, 2001 Apr, 12(4), 809 - 20 Sequence-specific interaction between the disintegrin domain of mouse ADAM 3 and murine eggs: role of beta1 integrin-associated proteins CD9, CD81, and CD98; Takahashi Y et al.; ADAM 3 is a sperm surface glycoprotein that has been implicated in sperm-egg adhesion . Because little is known about the adhesive activity of ADAMs, we investigated the interaction of ADAM 3 disintegrin domains, made in bacteria and in insect cells, with murine eggs . Both recombinant proteins inhibited sperm-egg binding and fusion with potencies similar to that which we recently reported for the ADAM 2 disintegrin domain . Alanine scanning mutagenesis revealed a critical importance for the glutamine at position 7 of the disintegrin loop . Fluorescent beads coated with the ADAM 3 disintegrin domain bound to the egg surface . Bead binding was inhibited by an authentic, but not by a scrambled, peptide analog of the disintegrin loop . Bead binding was also inhibited by the function-blocking anti-alpha6 monoclonal antibody (mAb) GoH3, but not by a nonfunction blocking anti-alpha6 mAb, or by mAbs against either the alphav or beta3 integrin subunits . We also present evidence that in addition to the tetraspanin CD9, two other beta1-integrin-associated proteins, the tetraspanin CD81 as well as the single pass transmembrane protein CD98 are expressed on murine eggs . Antibodies to CD9 and CD98 inhibited in vitro fertilization and binding of the ADAM 3 disintegrin domain . Our findings are discussed in terms of the involvement of multiple sperm ADAMs and multiple egg beta1 integrin-associated proteins in sperm-egg binding and fusion . We propose that an egg surface "tetraspan web" facilitates fertilization and that it may do so by fostering ADAM-integrin interactions. Biochemistry, 2001 Apr 17, 40(15), 4679 - 85 Roles of amino acid residues near the chromophore of photoactive yellow protein; Imamoto Y et al.; To investigate the roles of amino acid residues around the chromophore in photoactive yellow protein (PYP), new mutants, Y42A, E46A, and T50A were prepared . Their spectroscopic properties were compared with those of wild-type, Y42F, E46Q, T50V, R52Q, and E46Q/T50V, which were previously prepared and specified . The absorption maxima of Y42A, E46A, and T50A were observed at 438, 469, and 454 nm, respectively . The results of pH titration for the chromophore demonstrated that the chromophore of PYP mutant, like the wild-type, was protonated and bleached under acidic conditions . The red-shifts of the absorption maxima in mutants tended toward a pK(a) increase . Mutation at Glu46 induced remarkable shifts in the absorption maxima and pK(a) . The extinction coefficients were increased in proportion to the absorption maxima, whereas the oscillator strengths were constant . PYP mutants that conserved Tyr42 were in the pH-dependent equilibrium between two states (yellow and colorless forms) . However, Y42A and Y42F were in the pH-independent equilibrium between additional intermediate state(s) at around neutral pH, in which yellow form was dominant in Y42F whereas the other was dominant in Y42A . These findings suggest that Tyr42 acts as the hinge of the protein, and the bulk as well as the hydroxyl group of Tyr42 controls the protein conformation . In all mutants, absorbance at 450 nm was decreased upon flash irradiation and afterwards recovered on a millisecond time scale . However, absorbance at 340--370 nm was increased vice versa, indicating that the long-lived near-UV intermediates are formed from mutants, as in the case of wild-type . The lifetime changes with mutation suggest the regulation of proton movement through a hydrogen-bonding network. Bioorg Med Chem Lett, 2001 Apr 9, 11(7), 895 - 8 The discovery of a potent, intracellular, orally bioavailable, long duration inhibitor of human neutrophil elastase--GW311616A a development candidate; Macdonald SJ et al.; The discovery of a potent intracellular inhibitor of human neutrophil elastase which is orally active and has a long duration of action is described . The pharmacodynamic and pharmacokinetic properties of a trans-lactam development candidate, GW311616A, are described. Fresenius J Anal Chem, 2001 Feb, 369(3-4), 258 - 66 Luminescent proteins from Aequorea victoria: applications in drug discovery and in high throughput analysis; Deo SK et al.; Recent progress in generating a vast number of drug targets through genomics and large compound libraries through combinatorial chemistry have stimulated advancements in drug discovery through the development of new high throughput screening (HTS) methods . Automation and HTS techniques are also highly desired in fields such as clinical diagnostics . Luminescence-based assays have emerged as an alternative to radiolabel-based assays in HTS as they approach the sensitivity of radioactive detection along with ease of operation, which makes them amenable to miniaturization . Luminescent proteins provide the advantage of reduced reagent and operating costs because they can be produced in unlimited amounts through the use of genetic engineering tools . In that regard, the use of two naturally occurring and recombinantly produced luminescent proteins from the jellyfish Aequorea victoria, namely, aequorin and the green fluorescent protein (GFP), has attracted attention in a number of analytical applications in diverse research areas . Aequorin is naturally bioluminescent and has therefore, virtually no associated background signal, which allows its detection down to attomole levels . GFP has become the reporter of choice in a variety of applications given that it is an autofluorescent protein that does not require addition of any co-factors for fluorescence emission . Furthermore, the generation of various mutants of GFP with differing luminescent and spectral properties has spurred additional interest in this protein . In this review, we focus on the use of aequorin and GFP in the development of highly sensitive assays that find applications in drug discovery and in high throughput analysis. Infect Immun, 2001 May, 69(5), 3181 - 9 Apoptotic signaling pathway activated by Helicobacter pylori infection and increase of apoptosis-inducing activity under serum-starved conditions; Shibayama K et al.; The enhanced gastric epithelial cell apoptosis observed during infection with Helicobacter pylori has been suggested to be of significance in the etiology of gastritis, peptic ulcers, and neoplasia . To investigate the cell death signaling induced by H . pylori infection, human gastric epithelial cells were incubated with H . pylori for up to 72 h . H . pylori infection induced the activation of caspase -8, -9, and -3 and the expression of the proapoptotic Bcl-2 family proteins Bad and Bid . The peak of the activity of the caspases occurred at 24 h . At this time, the inhibition of caspase-8 or -9 almost completely suppressed H . pylori-induced apoptosis . Inhibition of caspase-8 suppressed the expression of Bad and Bid and the subsequent activation of caspase-9 and -3 . These observations indicate that H . pylori induces apoptosis through a pathway involving the sequential induction of apical caspase-8 activity, the proapoptotic proteins Bad and Bid, caspase-9 activity, and effector caspase-3 activity . Activation of the pathway was independent of CagA or vacuolating toxin . A membrane fraction of H . pylori was sufficient to activate this pathway, and treatment with proteinase K eliminated the activity . Apoptotic activity of the membrane fraction was significantly increased by incubating the bacteria under serum-starved conditions for 24 h . These observations suggest that environmental conditions in the human stomach could induce H . pylori-mediated pathogenesis, leading to a variety of clinical outcomes. Vet Immunol Immunopathol, 2001 Feb 10, 78(3-4), 219 - 29 Characterization of caprine interleukin-4; Snekvik KR et al.; Caprine interleukin-4 (IL-4) cDNA was cloned from RNA of mitogen stimulated goat peripheral blood mononuclear cells utilizing reverse transcriptase-polymerase chain reaction . The sequence of caprine IL-4 cDNA corresponds to a 535 nucleotide mRNA with 5'- and 3'-untranslated regions and a 405 nucleotide open reading frame, the first 66 nucleotides of which encode a putative signal peptide . Mature IL-4 is a 12.8kDa protein containing six cysteine residues and two potential N-linked glycosylation sites and is highly homologous with other ruminant IL-4 . The predicted molecular mass of mature unglycosylated IL-4 was confirmed by western blot of recombinant caprine IL-4 expressed in bacteria with a monoclonal antibody against a carboxyterminal peptide derived from the predicted amino acid sequence of bovine IL-4 . Eukaryotic expression plasmids containing caprine IL-4 cDNA were used to characterize recombinant IL-4 . Transcription of IL-4 mRNA was confirmed by transfection of COS-7 and goat synovial membrane cells, and recombinant IL-4 produced by stably transfected L929 cells inhibited inducible nitric oxide synthase in macrophages . Genetic immunization of mice with a caprine IL-4 cDNA expression plasmid induced antibodies against recombinant caprine IL-4 produced in bacteria. J Mol Biol, 2001 Apr 13, 307(5), 1395 - 410 Structure, function, and dynamics of the dimerization and DNA-binding domain of oncogenic transcription factor v-Myc; Fieber W et al.; The protein product (c-Myc) of the protooncogene c-myc is a transcriptional regulator playing a key role in cellular growth, differentiation, and apoptosis . Deregulated myc genes, like the transduced retroviral v-myc allele, are oncogenic and cause cell transformation . The C-terminal bHLHZip domain of v-Myc, encompassing protein dimerization (helix-loop-helix, leucine zipper) and DNA contact (basic region) surfaces, was expressed in bacteria as a highly soluble p15(v-myc )recombinant protein . Dissociation constants (K(d)) for the heterodimer formed with the recombinant bHLHZip domain of the Myc binding partner Max (p14(max)) and for the Myc-Max-DNA complex were estimated using circular dichroism (CD) spectroscopy and quantitative electrophoretic mobility shift assay (EMSA) . Multi-dimensional NMR spectroscopy was used to characterize the solution structural and dynamic properties of the v-Myc bHLHZip domain . Significant secondary chemical shifts indicate the presence of two separated alpha-helical regions . The C-terminal leucine zipper region forms a compact alpha-helix, while the N-terminal basic region exhibits conformational averaging with substantial alpha-helical content . Both helices lack stabilizing tertiary side-chain interactions and represent exceptional examples for loosely coupled secondary structural segments in a native protein . These results and CD thermal denaturation data indicate a monomeric state of the v-Myc bHLHZip domain . The (15)N relaxation data revealed backbone mobilities which corroborate the existence of a partially folded state, and suggest a "beads-on-a-string" motional behaviour of the v-Myc bHLHZip domain in solution . The preformation of alpha-helical regions was confirmed by CD thermal denaturation studies, and quantification of the entropy changes caused by the hydrophobic effect and the reduction of conformational entropy upon protein dimerization . The restricted conformational space of the v-Myc bHLHZip domain reduces the entropy penalty associated with heterodimerization and allows rapid and accurate recognition by the authentic Myc binding partner Max . Transplantation, 2001 Mar 15, 71(5), 611 - 8 Bone marrow transplant conditioning intensified with liposomal clodronate to eliminate residual host antigen presenting cells fails to ameliorate GVHD and increases PERI-BMT mortality; Everse LA et al.; BACKGROUND: Graft versus host disease (GVHD) mediated by allogeneic donor T cells may be initiated and/or exacerbated by residual host antigen presenting cells (APC) which survive the transplant conditioning regimen . We examined whether the depletion of hepatic and splenic APC could reduce the severity of hepatic GVHD after bone marrow transplantation (BMT) . METHODS: Recipient mice were depleted for hepatic and splenic phagocytic APCs by i.v . injection of clodronate- (dichloromethylene diphosphonate) containing liposomes before fully allogeneic or MHC-matched, minor Ag-mismatched BMT . Severity of hepatic GVHD was scored on histological sections 2, 3, 4, or 9 weeks after BMT . RESULTS: No differences in the severity of GVHD were observed between APC-depleted mice and control mice . APC-depleted mice had increased peritransplant mortality due to sepsis . Bacterial clearance assays showed that APC-depleted mice were unable to efficiently clear bacteria, although nondepleted, transplanted mice were able to clear bacteria as quickly as naive control mice . CONCLUSIONS: Residual host phagocytic APC do not appear to play a role in the induction of GVHD after BMT . They are, however, essential for prevention of sepsis in the transplant host. Plant Mol Biol, 2001 Feb, 45(3), 341 - 52 The low-temperature- and salt-induced RCI2A gene of Arabidopsis complements the sodium sensitivity caused by a deletion of the homologous yeast gene SNA1; Nylander M et al.; Two closely related, tandemly arranged, low-temperature- and salt-induced Arabidopsis genes, corresponding to the previously isolated cDNAs RCI2A and RCI2B, were isolated and characterized . The RCI2A transcript accumulated primarily in response to low temperature or high salinity, and to a lesser extent in response to ABA treatment or water deficit stress . The RCI2B transcript was present at much lower levels than RCI2A, and could only be detected by reverse transcription-PCR amplification . The predicted 6 kDa RCI2 proteins are highly hydrophobic and contain two putative membrane-spanning regions . The polypeptides exhibit extensive similarity to deduced low-temperature- and/or salt-induced proteins from barley, wheat grass and strawberry, and to predicted proteins from bacteria, fungi, nematodes and yeast . Interestingly, we found that a deletion of the RCI2 homologous gene, SNA1 (YRD276c), in yeast causes a salt-sensitive phenotype . This effect is specific for sodium, since no growth defect was observed for the sna1 mutant on 1.7 M sorbitol, 1 M KCl or 0.6 M LiCl . Finally, we found that the Arabidopsis RCI2A cDNA can complement the sna1 mutant when expressed in yeast, indicating that the plant and yeast proteins have similar functions during high salt stress. Immunol Rev, 2001 Feb, 179, 57 - 60 Regulation of human intestinal mast cells by stem cell factor and IL-4; Lorentz A et al.; Mature human mast cells are tissue-residing, key effector cells of immediate allergic reactions . Moreover, mast cells have been recognized as a potent cellular source of multiple cytokines, suggesting an important role in immunoregulation and host defense . Here, we report on the regulation of mature human mast cells isolated from intestinal tissues by stem cell factor (SCF) and interleukin (IL)-4 . SCF is substantially necessary for mast cell survival and induces marginal mast cell proliferation in vitro, whereas IL-4 by itself has no effects on mast cell survival or proliferation . Most interestingly, in synergy with SCF, IL-4 strongly enhances mast cell proliferation . In the presence of SCF, mast cells predominantly produce pro-inflammatory cytokines including tumor necrosis factor (TNF)-alpha, IL-1beta, IL-6, IL-8, IL-16, and IL-18 . Addition of IL-4 to the culture medium induces the expression of Th2-type cytokines (IL-3, IL-5 and IL-13), and a downregulation of pro-inflammatory cytokines, namely IL-6 . Furthermore, SCF by itself supports the predominance of the tryptase/chymase double-positive mast cell subtype MCTC whereas the addition of IL-4 supports the chymase negative MCT subtype . In conclusion, SCF may primarily regulate resident mast cell survival, whereas IL-4 may promote local proliferation of mast cells and their expression of Th2-type cytokines. J Gastroenterol, 2001 Mar, 36(3), 153 - 7 Ecabet sodium inhibits the ability of Helicobacter pylori to induce neutrophil production of reactive oxygen species and interleukin-8; Shimoyama T et al.; The pathogenic role of human neutrophils has been implicated in Helicobacter pylori-associated gastritis . Ecabet sodium, a locally acting antiulcer drug, has anti-H . pylori actions . The aim of this study was to examine the effects of ecabet on the ability of H . pylori to stimulate human neutrophils . H . pylori were added to 1 x 10(5) neutrophils and incubated for 30 min in the presence of ecabet . Bacterial suspensions which had been incubated with ecabet for 30 min were also used to stimulate neutrophils . The intracellular production of reactive oxygen species was measured with a FACScan . Bacterial suspensions were also added to neutrophils in the presence of ecabet and incubated at 37 degrees C for 12 h to measure interleukin (IL)-8 production by enzyme-linked immunosorbent assay . The mean fluorescence intensity was found to be attenuated dose-dependently by ecabet (P < 0.01) . Ecabet also inhibited IL-8 production by neutrophils in a dose-dependent manner (P < 0.001) . Bacteria with prior incubation with ecabet induced significantly lower IL-8 production than those without this incubation (P < 0.05) . Ecabet sodium has preventive effects on the ability of H . pylori to stimulate human neutrophils . It may lead to reduced gastritis activity and decreased oxidative damage of the gastric mucosa in H . pylori-associated gastritis. Rapid Commun Mass Spectrom, 2001, 15(6), 393 - 400 Development of a method based on accelerated solvent extraction and liquid chromatography/mass spectrometry for determination of arylphenoxypropionic herbicides in soil; Marchese S et al.; A sensitive and specific analytical procedure for determining arylphenoxypropionic herbicides in soil samples, using Ionspray ionization (ISI) liquid chromatography/mass spectrometry (LC/MS), is presented . Arylphenoxypropionic acids are a new class of herbicides used for selective removal of most grass species from any non-grass crop, commercialized as herbicide esters . Previous studies have shown that the esters undergo fast hydrolysis in the presence of vegetable tissues and soil bacteria, yelding the corresponding free acid . The feasibility of rapidly extracting arylphenoxypropionic herbicides from soil by accelerated solvent extraction (ASE) techniques was evaluated . Four different soil samples were fortified with target compounds at levels of 5 and 20 ng/g by following a procedure able to mimic weathered soils . Herbicides were extracted by a methanol/water (80:20 v/v) solution (0.12 M) of NaCl at 90 degrees C . After clean-up using graphitized carbon black (GCB) as absorbent, the extract was analyzed by HPLC/ISI-MS . The effect of concentration of acid in the mobile phase on the response of ISI-MS was investigated . The effects of varying the orifice plate voltage on the production of diagnostic fragment ions, and on the response of the MS detector, were also investigated . The ISI-MS response was linearly related to the amounts of analytes injected between 1 and 200 ng . The limit of detection (signal-to-noise ratio = 3) of the method for the pesticides in soil samples was estimated to be less than 1 ng/g. Am J Pathol, 2001 Apr, 158(4), 1391 - 8 Elevated proportion of natural killer T cells in periodontitis lesions: a common feature of chronic inflammatory diseases; Yamazaki K et al.; Although periodontitis is a chronic inflammatory disease caused by a group of so-called periodontopathic bacteria, autoimmune mechanisms have also been implicated in the disease process . Recently, a unique subset of lymphocytes designated natural killer (NK) T cells expressing the Valpha24JalphaQ invariant T cell receptor (TCR) has been reported to have a regulatory role in certain autoimmune diseases . Therefore, we investigated the proportion of the invariant Valpha24JalphaQ TCR within the Valpha24 T cell population in periodontitis lesions and gingivitis lesions using single-strand conformation polymorphism methodology . NK T cells were identified with a specific JalphaQ probe whereas the total Valpha24 TCR was identified using an internal Calpha probe . NK T cells were a significant proportion of the total Valpha24 population both in periodontitis lesions and to a lesser extent in gingivitis lesions but not in the peripheral blood of either periodontitis patients or nondiseased controls . Using immunohistochemistry, some of Valpha24(+) cells in the periodontitis lesions seemed to associate with CD1d(+) cells, which are specific antigen-presenting cells for NK T cells . Although the mechanism underlying the elevation of NK T cells in periodontitis and in gingivitis lesions remains unclear, it can be postulated that NK T cells are recruited to a play regulatory role in the immune response to bacterial infection. Comp Biochem Physiol B Biochem Mol Biol, 2001 Apr, 128(4), 709 - 18 Protein purification, cDNA cloning and gene expression of lysozyme from eri-silkworm, Samia cynthia ricini; Fujimoto S et al.; Lysozyme was isolated from immunized hemolymph of Samia cynthia ricini larvae by heat treatment, cation exchange and reverse-phase chromatography . A cDNA encoding lysozyme was cloned by screening the cDNA library from immunized fat body using, as a probe, a DNA fragment obtained by PCR-based differential display method . The deduced amino acid sequence showed high homology with other chicken-type lysozymes . The calculated molecular mass of the mature peptide was 13785, which agreed precisely with that obtained by MALDI-TOF mass spectrometry of the isolated protein . The lysozyme transcripts were detected at a significant level in naive fat body, and the level increased 5-10-fold upon injection of the larvae with UV-killed bacteria or peptidoglycan. Gene, 2001 Mar 21, 266(1-2), 147 - 53 Simple and efficient vectors for retrofitting BACs and PACs with mammalian neoR and EGFP marker genes; Kaname T et al.; Bacterial artificial chromosomes (BACs) and P1 artificial chromosomes (PACs) are widely used to investigate the functions of genes and genomes in mammalian cells in vitro and in vivo . We have developed a series of vectors which can simply and efficiently be retrofitted onto BACs or PACs . These vectors carry a neoR gene for selection in cells in tissue culture, including ES cells, and also an EGFP gene driven by the strong CAG promoter for quick detection of the DNA in cells . All the plasmids are retrofitted using the loxP site and Cre recombinase and some carry the gamma origin of plasmid R6K which does not function in commonly used bacteria such as DH10B . Retrofitting of PACs and BACs carrying alphoid DNA was very efficient with almost no rearrangement of the highly repetitive alphoid DNA . Following transfer into HT1080 cells and mouse oocytes in tissue culture the DNA could easily be monitored by the EGFP fluorescence. J Hosp Infect, 2001 Apr, 47(4), 251 - 6 Surgical face masks in the operating theatre: re-examining the evidence; Romney MG; In most modern hospitals, no one is allowed to enter the operating theatre without wearing a surgical face mask . The practice of wearing masks is believed to minimize the transmission of oro- and nasopharyngeal bacteria from operating theatre staff to patients' wounds, thereby decreasing the likelihood of postoperative surgical site infections . In this era of cost-restraints, shrinking hospital budgets, and evidence-based medicine, many health care professionals have begun to re-examine traditional infection control practices . Over the past decade, studies challenging the accepted dogma of surgical face mask usage have been published . Masks that function as protective barriers are another emerging issue . Due to a greater awareness of HIV and other blood-borne viruses, masks are taking on a greater role in protecting health care workers from potentially infectious blood and body fluids . The purpose of this review is to evaluate the latest evidence for and against routine use of surgical face masks in the operating theatre . Skeletal Radiol, 2001 Jan, 30(1), 57 - 9 Unusual facet cyst containing struvite and hydroxyapatite; Grantham M et al.; This case report describes a patient with severe back pain and radiculopathy . She was found to have a facet cyst within the lumbar spine that appeared to contain calcium on MRI and CT . Upon aspiration the cyst was found to contain calcium ammonium phosphate (struvite) and calcium phosphate (hydroxyapatite) . Ammonia production in the presence of urease-producing bacteria is responsible for the production of struvite in the human body . We postulate that there was a prior infection of the facet with urease-producing bacteria, thus accounting for the production of the struvite within the facet cyst. J Med Microbiol, 2001 Apr, 50(4), 320 - 9 Involvement of the heparan sulphate-binding proteins of Helicobacter pylori in its adherence to HeLa S3 and Kato III cell lines; Guzman-Murillo MA et al.; To determine whether Helicobacter pylori heparan sulphate-binding proteins (HSBPs) are involved in the adherence of H . pylori to HeLa and Kato III cells, monolayers were pre-incubated with various preparations and concentrations of H . pylori HSBPs at 37 degrees C, washed and then challenged with bacteria . HSBPs did not prevent but enhanced H . pylori adherence . However, challenging cultured cells with H . pylori previously incubated with rabbit anti-HSBP IgG resulted in significant inhibition of bacterial adherence . These data demonstrate that the extracellular HSBP plays an important role in promoting H . pylori attachment to Kato III and HeLa S3 cells, that adhesion of H . pylori to Kato III and HeLa S3 cells is promoted by the presence of the 71.5-kDa extracellular HSBP and that rabbit polyclonal antibodies against this HSBP can inhibit adhesion of H . pylori to the cultured cell lines and detach cell-bound H . pylori. J Periodontol, 2001 Feb, 72(2), 146 - 51 Systemic neutrophil response resulting from dental plaque accumulation; Kowolik MJ et al.; BACKGROUND: There is considerable current interest in putative relationships between oral and systemic diseases . Since the host response to oral bacteria may be the critical link in this association, our hypothesis was that dental plaque accumulation in healthy subjects would elicit a systemic inflammatory response . METHODS: Twenty-three healthy subjects, aged 18 to 25, participated in a 4-phase study . An initial hygiene phase was followed by a 21-day experimental phase (the so-called experimental gingivitis model) in which subjects refrained from all oral hygiene practices, thus permitting the accumulation of bacterial plaque . At days 0, 7, and 21 total and differential peripheral white blood cell (wbc) counts, together with full mouth plaque and gingivitis scores, were recorded . Following a 28-day recovery phase, in which normal oral hygiene practices were resumed, subjects entered the final 21-day control phase which mirrored the experimental phase but with subjects maintaining normal oral hygiene practices . RESULTS: The experimental model performed as anticipated with a correlation between plaque and gingivitis scores of 0.95, also reflecting subject compliance . Total wbc and neutrophil counts increased during the experimental phase . Furthermore, comparison of neutrophil counts between the experimental and control phases demonstrated a significantly higher cell count for the experimental phase on both days 7 and 21 (P= 0.0301 and 0.009, respectively) . For total wbc, this was significant on day 21 (P= 0.0262) . CONCLUSION: The results of this study support the hypothesis that the accumulation of dental plaque can result in a measurable systemic inflammatory response, providing further in vivo data to support a mechanistic relationship between oral and systemic pathology. RN, 2001 Mar, 64(3), 28 - 31; quiz 32 The right way to do blood cultures; Ernst DJ; Blood samples contaminated by skin-surface bacteria can increase hospital stays, cause the needless use of antibiotics, and cost thousands of dollars per incident . A few simple steps can reduce contamination and save patients and healthcare providers money, time, and heartache. Polim Med, 2000, 30(3-4), 89 - 98 Evaluation of the healing process after implantation of synthetic material called Tegmentum consisting of polyester mesh covered with polyurethane in rat peritoneum; Fila M et al.; Polyester meshes are one of the synthetic prosthetic materials widely applied for the reconstruction of abdominal layers in child and neonatal surgery, cardiosurgery or vascular surgery . Because of the lack or insufficiency of the own, natural material multiple researches are performed in order to find the best material able to replace natural tissue . The most suitable prosthesis ought to be: sterile, non-toxic, soft, flexible, elastic, not very stretchy, durable, easy to operate (cutting, suturing); to create desired shape required while reconstracting, totally resistant to contagion, without any complication after being implanted . Since early seventies many scientists following above requirements have carried out researches to create the most similar prosthesis morphologically and functionally to human tissue but still there is no such an ideal material on the market . Polyester with its derivatives is one of the most commonly applied synthetic substances in surgery . The type of biomaterial highly depends on its purpose e.g.: for vascular prosthesis materials that are resorptive in organism and support tightness (gelatine, albumin, collagen); while for temporal replacement abdominal layers with prosthesis the most suitable is impervious to systemic fluids bacteria etc material, that prevents penetrating the surface of prosthesis by surrounding tissues, adhesions with intestinum or other organs . Tegmentum is a material that posses these features . This is a polyester mesh covered with polyurethane on one side produced by Tricomed S.A . (Lodz) . Tegmentum has already been applied to neonates with congenital eventration in Child and Neonatal Ward in the "Szpital Pomnik Matki Polki" hospital in Lodz. Proc Natl Acad Sci U S A, 2001 Apr 10, 98(8), 4299 - 304 Epub 2001 Apr 03. On the absorbance changes in the photocycle of the photoactive yellow protein: a quantum-chemical analysis; Molina V et al.; Spectral changes in the photocycle of the photoactive yellow protein (PYP) are investigated by using ab initio multiconfigurational second-order perturbation theory at the available structures experimentally determined . Using the dark ground-state crystal structure {Genick, U . K., Soltis, S . M., Kuhn, P., Canestrelli, I . L . & Getzoff, E . D . (1998) Nature (London) 392, 206-209}, the pipi* transition to the lowest excited state is related to the typical blue-light absorption observed at 446 nm . The different nature of the second excited state (npi*) is consistent with the alternative route detected at 395-nm excitation . The results suggest the low-temperature photoproduct PYP(HL) as the most plausible candidate for the assignment of the cryogenically trapped early intermediate (Genick et al.) . We cannot establish, however, a successful correspondence between the theoretical spectrum for the nanosecond time-resolved x-ray structure {Perman, B., Srajer, V., Ren, Z., Teng, T., Pradervand, C., et al . (1998) Science 279, 1946-1950} and any of the spectroscopic photoproducts known up to date . It is fully confirmed that the colorless light-activated intermediate recorded by millisecond time-resolved crystallography {Genick, U . K., Borgstahl, G . E . O., Ng, K., Ren, Z., Pradervand, C., et al . (1997) Science 275, 1471-1475} is protonated, nicely matching the spectroscopic features of the photoproduct PYP(M) . The overall contribution demonstrates that a combined analysis of high-level theoretical results and experimental data can be of great value to perform assignments of detected intermediates in a photocycle. Glycobiology, 2001 Feb, 11(2), 11R - 18R Biochemical engineering of the N-acyl side chain of sialic acid: biological implications; Keppler OT et al.; N-Acetylneuraminic acid is the most prominent sialic acid in eukaryotes . The structural diversity of sialic acid is exploited by viruses, bacteria, and toxins and by the sialoglycoproteins and sialoglycolipids involved in cell-cell recognition in their highly specific recognition and binding to cellular receptors . The physiological precursor of all sialic acids is N-acetyl D-mannosamine (ManNAc) . By recent findings it could be shown that synthetic N-acyl-modified D-mannosamines can be taken up by cells and efficiently metabolized to the respective N-acyl-modified neuraminic acids in vitro and in vivo . Successfully employed D-mannosamines with modified N-acyl side chains include N-propanoyl- (ManNProp), N-butanoyl- (ManNBut)-, N-pentanoyl- (ManNPent), N-hexanoyl- (ManNHex), N-crotonoyl- (ManNCrot), N-levulinoyl- (ManNLev), N-glycolyl- (ManNGc), and N-azidoacetyl D-mannosamine (ManNAc-azido) . All of these compounds are metabolized by the promiscuous sialic acid biosynthetic pathway and are incorporated into cell surface sialoglycoconjugates replacing in a cell type-specific manner 10-85% of normal sialic acids . Application of these compounds to different biological systems has revealed important and unexpected functions of the N-acyl side chain of sialic acids, including its crucial role for the interaction of different viruses with their sialylated host cell receptors . Also, treatment with ManNProp, which contains only one additional methylene group compared to the physiological precursor ManNAc, induced proliferation of astrocytes, microglia, and peripheral T-lymphocytes . Unique, chemically reactive ketone and azido groups can be introduced biosynthetically into cell surface sialoglycans using N-acyl-modified sialic acid precursors, a process offering a variety of applications including the generation of artificial cellular receptors for viral gene delivery . This group of novel sialic acid precursors enabled studies on sialic acid modifications on the surface of living cells and has improved our understanding of carbohydrate receptors in their native environment . The biochemical engineering of the side chain of sialic acid offers new tools to study its biological relevance and to exploit it as a tag for therapeutic and diagnostic applications. J Dairy Sci, 2001 Mar, 84(3), 680 - 90 Effect of dietary lipid source on conjugated linoleic acid concentrations in milk fat; Chouinard PY et al.; Conjugated linoleic acids (CLA) found in ruminant milk fat are a byproduct of incomplete biohydrogenation of lipids by ruminal bacteria . We examined the effect of different dietary fat supplements and processing methods on CLA . In trial 1, dietary supplements of Ca salts of fatty acids from canola oil, soybean oil, and linseed oil increased CLA content of milk fat by three- to fivefold over the control diet . Trials 2 and 3 examined the effect of processing methods for heat treatment of full fat soybeans . In trial 2, extrusion, micronizing, and roasting resulted in two- to threefold greater concentrations of CLA in milk fat than the control diet (raw ground soybeans) . In trial 3, different temperatures of extrusion (120, 130, and 140 degrees C) increased the CLA content of milk fat to a similar extent; CLA averaged 19.9 mg/g of fatty acids for the extrusion treatments compared with 4.2 mg/g of fatty acids for the control diet (raw ground soybeans) . Fish oil (200 and 400 ml/d) was examined in trial 4 and both levels resulted in CLA concentrations in milk fat that were about threefold greater than the control diet . In trial 5, grain and silage from a high oil corn hybrid increased the CLA content of milk fat; however, responses were modest with the CLA concentration (mg/g of fatty acids) averaging 4.6 and 2.8 for diets with high oil hybrid and normal hybrid, respectively . Similarly, dietary supplements of animal fat byproducts (tallow plus yellow grease; trial 6) resulted in modest increases in the CLA content of milk fat . Overall, several dietary manipulations involving lipid sources and processing methods were identified that allow for a marked increase in the conjugated linoleic acid content of milk fat. J Environ Qual, 2001 Mar-Apr, 30(2), 403 - 10 Solid-state nitrogen-15 nuclear magnetic resonance analysis of biologically reduced 2,4,6-trinitrotoluene in a soil slurry remediation; Knicker H et al.; Soil contaminated with 2,4,6-trinitrotoluene (TNT) and spiked with {14C}- and {15N3}-TNT was subjected to an anaerobic-aerobic soil slurry treatment and subsequently analyzed by radiocounting and solid-state 15N nuclear magnetic resonance (NMR) spectroscopy . This treatment led to a complete disappearance of extractable radioactivity originating from TNT and almost all of the radioactivity was recovered in the insoluble soil fraction . As revealed by solid-state 15N NMR, a major fraction of partially reduced metabolites of TNT was immobilized into the soil during the early stage of the anaerobic treatment, although some of the compounds (i.e., aminodinitrotoluenes and azoxy compounds) were extractable by methanol . Considerable 15N intensity was assigned to condensation products of TNT metabolites . A smaller signal indicated the formation of azoxy N . This signal and the signal for nitro groups were not observed at the end of the anaerobic phase, revealing further reduction and/or transformation of their corresponding compounds . An increase of the relative proportion of the condensation products occurred with increasing anaerobic incubation . Aerobic incubation resulted in a further decrease of aromatic amines, presumably due to oxidative transformations or their involvement in further condensation reactions . The results of the study demonstrate that the anaerobic-aerobic soil slurry treatment represents an efficient strategy for immobilizing reduced TNT in soils. J Environ Qual, 2001 Mar-Apr, 30(2), 369 - 76 Methane oxidation in two Swedish landfill covers measured with carbon-13 to carbon-12 isotope ratios; Borjesson G et al.; The release of methane (CH4) from landfills to the atmosphere and the oxidation of CH4 in the cover soils were quantified with static chambers and a 13C-isotope technique on two landfills in Sweden . One of the landfills had been closed and covered 17 years before this investigation while the other was recently covered . On both landfills, the tops of the landfills were compared with the sloping parts in the summer and winter . Emitted CH4, captured in chambers, was significantly enriched in 13C during summer compared with winter (P < 0.0001), and was enriched relative to anaerobic-zone methane . The difference between emitted and anaerobic zone delta 13C-CH4 was used to estimate soil methane oxidation . In summer, these differences ranged from 9 to 26@1000, and CH4 oxidation was estimated to be between 41 and 50% of the produced CH4 in the new landfill, and between 60 and 94% in the old landfill . In winter, when soil temperature was below 0 degree C, no difference in delta 13C was observed between emitted and anaerobic-zone CH4, suggesting that there was no soil oxidation . The temperature effect shown in this experiment suggests that there may be both seasonal and latitudinal differences in the importance of landfill CH4 oxidation . Finally the isotopic fractionation factor (alpha) varied from 1.023 to 1.038 and was temperature dependent, increasing at colder temperatures . Methanotrophic bacteria appeared to have high growth efficiencies and the majority of the methane consumed in incubations did not result in immediate CO2 production. Traffic, 2001 Apr, 2(4), 245 - 51 Multiple pathways used for the targeting of thylakoid proteins in chloroplasts; Robinson C et al.; The assembly of the chloroplast thylakoid membrane requires the import of numerous proteins from the cytosol and their targeting into or across the thylakoid membrane . It is now clear that multiple pathways are involved in the thylakoid-targeting stages, depending on the type of protein substrate . Two very different pathways are used by thylakoid lumen proteins; one is the Sec pathway which has been well-characterised in bacteria, and which involves the threading of the substrate through a narrow channel . In contrast, the more recently characterised twin-arginine translocation (Tat) system is able to translocate fully folded proteins across this membrane . Recent advances on bacterial Tat systems shed further light on the structure and function of this system . Membrane proteins, on the other hand, use two further pathways . One is the signal recognition particle-dependent pathway, involving a complex interplay between many different factors, whereas other proteins insert without the assistance of any known apparatus . This article reviews advances in the study of these pathways and considers the rationale behind the surprising complexity. Aliment Pharmacol Ther, 2001 Apr, 15(4), 543 - 9 Relationship between lactose digestion, gastrointestinal transit time and symptoms in lactose malabsorbers after dairy consumption; Labayen I et al.; BACKGROUND: The relationship of symptoms with objective measurements, as well as some of the mechanisms involved in lactose tolerance after yoghurt consumption, remain unclear . METHODS: The trial had a double-blind design in which 22 lactose malabsorbers received 25 g daily lactose in fresh (living bacteria > 108 cfu/g) yoghurt or heated (< 102 cfu/g) yoghurt for 15 days, followed by a cross-over (15 days) after a wash-out period (14 days) . The lactose digestion was determined by the breath H2 test, the gastric emptying (GE) with a 13C-acetate breath test and the revealed transit time (OCTT) by 15N-lactose-ureide test . Subjects reported their gastrointestinal symptoms (GIS) in a validated questionnaire . RESULTS: Breath H2 test indicated more effective lactose digestion after fresh yoghurt intake . The OCTT was shorter after heated yoghurt ingestion as compared with the fresh . There was lower severity of GIS (P < 0.05) after fresh yoghurt intake, and this showed an inverse correlation with OCTT (P < 0.05) . CONCLUSIONS: Delayed orocoecal transit time was associated with fewer gastrointestinal symptoms . The improved lactose digestion and tolerance of fresh yoghurt should be mainly attributed to the presence of living bacteria. Biochem J, 2001 Apr 15, 355(Pt 2), 499 - 507 GTP cyclohydrolase I mRNA: novel splice variants in the slime mould Physarum polycephalum and in human monocytes (THP-1) indicate conservation of mRNA processing; Golderer G et al.; GTP cyclohydrolase I (EC 3.5.4.16) is the first enzyme in the biosynthesis of tetrahydrobiopterin {(6R)-5,6,7,8-tetrahydro-L-biopterin, H(4)-biopterin} in mammals and of folic acid in bacteria . Here we have characterized the GTP cyclohydrolase I gene structure and two mRNA species from Physarum polycephalum, an acellular slime mould that synthesizes H(4)-biopterin and metabolites of the folic acid biosynthetic pathway . Its GTP cyclohydrolase I gene consists of seven exons, and the two GTP cyclohydrolase I cDNA species isolated from Physarum encode for proteins with 228 (25.7 kDa) and 195 (22.1 kDa) amino acids . Furthermore, we identified two previously undescribed mRNA species in interferon-gamma-treated human myelomonocytoma cells (THP-1) in addition to the cDNA coding for the fully functional 250-residue (27.9 kDa) protein, which is identical with that in human phaeochromocytoma cells . One of the new splice variants codes for a 233-residue (25.7 kDa) protein, whereas the other codes for the full-length protein but is alternatively spliced within the 3'-untranslated region . In heterologous expression, the shorter proteins of Physarum as well as of THP-1 cells identified here are degraded by proteolysis . Accordingly, only the 27.9 kDa protein was detectable in Western blots from THP-1 cell extracts . Quantification of GTP cyclohydrolase I mRNA species in different human cell types with and without cytokine treatment showed that in addition to the correct mRNA the two splice variants isolated here, as well as the two splice variants known from human liver, are strongly induced by cytokines in cell types with inducible GTP cyclohydrolase I (THP-1, dermal fibroblasts), but not in cell types with constitutive GTP cyclohydrolase I expression (SK-N-SH, Hep-G2) . As in human liver, splicing of the new mRNA variant found in THP-1 cells occurs at the boundary of exons 5 and 6 . Strikingly, the 195-residue protein from Physarum is alternatively spliced at a homologous position, i.e . at the boundary of exons 6 and 7 . Thus alternative splicing of GTP cyclohydrolase I at this position occurs in two species highly distant from each other in terms of evolution . It remains to be seen whether variant proteins encoded by alternatively spliced GTP cyclohydrolase I mRNA transcripts do occur in vivo and whether they participate in regulation of enzyme activity. Eur J Ophthalmol, 2001 Jan-Mar, 11(1), 31 - 6 Does surgical technique influence cataract surgery contamination? Koc F, Akcam Z, Kuruoglu S, Oge I, Gunaydin M. PURPOSE: To compare cataract surgery contamination rates in large-incision extracapsular cataract extraction (ECCE) and phacoemulsification (PE), we studied 65 cases prospectively . METHODS: Thirty-five cases were operated by large-incision ECCE (Group I) and 30 by PE (Group II) . Conjunctival swab cultures were taken immediately before surgery and anterior chamber aspirate was taken for culture upon completion of surgery for each case . RESULTS: Anterior chamber cultures were positive in 22.8% of the cases in group I and 23% in Group II . Frequencies of contamination in each group were no different (x2: 0.22, p>0.05) . When the contaminations were evaluated in relation to operating time, prolongation of the operating time raised the contamination rate in Group I (p<0.05) but not in Group II (p>0.05) . Silicone and PMMA intraocular lenses (IOL) were tested to see whether they had any additional risk of contamination . The frequencies of contaminated silicone IOL implanted cases (6/26) and contaminated PMMA IOL implanted cases (8/39) were similar (x2: 0.36, p>0 . 05) . CONCLUSIONS: Although the architecture of the incision and irrigation dynamics provided an advantage to the PE technique as the operating time became longer, routine PE was not superior to classical ECCE with respect to contamination when performed in the same circumstances . Prolonging the operating time raised the contamination rate in classical ECCE. Cad Saude Publica, 2001 Mar-Apr, 17(2), 385 - 96 {Chlamydia infection impact among native Indian groups of the Brazilian Amazon region}; Ishak MO et al.; Knowledge is limited on the spread of bacteria from genus Chlamydia in Brazil . This study included a sero-epidemiological survey of 2,086 samples from native Indian populations of the Brazilian Amazon region . Sera were screened using indirect immunofluorescence assay for detection of antibodies to C . trachomatis serotype L2, followed by microimmunofluorescence assay using fifteen C . trachomatis and C . pneumoniae serotypes as antigen substrates . Antibody prevalence was 48.6%, but there was a large prevalence range among the groups, including those that had never been challenged with the bacteria, as well as those in which almost all individuals had been infected . Titration of IgG antibodies and detection of specific IgM in high-titer samples showed the persistence of Chlamydia in 6.1% of the reactive individuals, who probably play an important role as reservoirs for dissemination of the bacteria . Specific seroreactivity to C . trachomatis showed the presence of serotypes A, B, Ba, D, E, G, H, I, and L1 in the geographic area surveyed . Furthermore, the survey showed that C . pneumoniae was also infecting these individuals . Both species may be involved in a significant human disease burden that merits further clarification. Science, 2001 Apr 20, 292(5516), 504 - 7 Epub 2001 Mar 29. Cooperation and competition in the evolution of ATP-producing pathways; Pfeiffer T et al.; Heterotrophic organisms generally face a trade-off between rate and yield of adenosine triphosphate (ATP) production . This trade-off may result in an evolutionary dilemma, because cells with a highe