Glia . 2005 Jan 18; {Epub ahead of print} Intermediate filament protein synemin is present in human reactive and malignant astrocytes and associates with ruffled membranes in astrocytoma cells; Jing R et al.; Synemin, a very unique type VI intermediate filament (IF) protein, exhibits alternative splice variants termed alpha and beta . Unlike other IF proteins, synemin binds to actin-associated proteins, including alpha-actinin, vinculin, and alpha-dystrobrevin . Our previous work has demonstrated the presence of synemin in differentiating astrocytes . In this study, we have examined the presence of synemin in human astrocytes under pathological conditions, using rabbit antibodies raised against the C-terminal domain of human synemin produced in bacteria . Western blotting shows that astrocytic tumors contain greater amounts of alpha-synemin than do normal brain tissues . These tumors also contain beta-synemin, which is not detectable in normal brain . Immunohistochemistry demonstrates that, while synemin is present in normal adult brain only in vascular smooth muscle cells, it is newly synthesized by reactive and neoplastic astrocytes . alpha- and beta-Synemins have also been detected by Western blotting and polymerase chain reaction in several human glioblastoma cell lines . In these cell lines, surprisingly, synemin is associated with ruffled membranes in addition to being distributed along the IF network . In ruffled membranes, synemin was found to co-localize with alpha-actinin . This unusual cellular localization for an IF protein is maintained after nocodazole-induced perinuclear coiling of the vimentin IF network . In addition, immunoprecipitation experiments demonstrate that synemin forms a complex with alpha-actinin in glioblastoma cells . Taken together with synemin localization within ruffled membranes, this finding suggests that synemin plays a role in motility of glioblastoma cells . (c) 2005 Wiley-Liss, Inc. J Intern Med, 2005 Feb, 257(2), 126 - 38 The immune system as the sixth sense; Blalock JE; Abstract . Blalock JE (University of Alabama, Birmingham, USA) . The immune system as the sixth sense (Minisymposium) . J Intern Med 2005; 257: 126-138.One of the truly remarkable discoveries in modern biology is the finding that the nervous system and immune system use a common chemical language for intra- and inter-system communication . This review will discuss some of the pivotal results that deciphered this chemical language . Specifically the nervous and immune systems produce a common set of peptide and nonpeptide neurotransmitters and cytokines that act on a common repertoire of receptors in the two systems . The paper will also review more recent studies that have delineated hardwired and humoral pathways for such bidirectional communication . This is discussed in the context of the idea that the sharing of ligands and receptors allows the immune system to serve as the sixth sense that notifies the nervous system of the presence of entities, such as viruses and bacteria, that are imperceptible to the classic senses . Lastly, this review will suggest ways to apply the newfound knowledge of the sixth sense to understand a placebo effect and to treate human disease. Blood Purif, 2004, 22 Suppl 2, 26 - 9 Use of Ultrapure Dialysate in Reduction of Chronic Inflammation during Hemodialysis; Arizono K et al.; Chronic inflammation contributes to the pathogenesis of several complications of hemodialysis therapy . It is thought that backfiltration of bacteria-derived contaminations during dialysis may induce a chronic inflammatory state . High-sensitivity C-reactive protein (hs-CRP) is one of the tools which can take a hold on such a chronic inflammatory condition . We examined the effect of ultrapure dialysate which contributes to chronic inflammation with hs-CRP and tried to reduce endotoxin (ET) levels at the end of the dialysate from 70 EU/l to <1.0 EU/l (ultrapure dialysate) . Other dialysis conditions, except ET level, were fixed . We investigated the hs-CRP of 23 patients receiving regular dialysis before the use of ultrapure dialysate and 1 year after use of it prospectively . The data showed a significant decrease in the median value of the hs-CRP from 0.16 to 0.07 mg/dl (p < 0.05) . The value of serum beta(2)-microglobulin decreased from 33.2 to 28.4 mg/dl (p < 0.01) and the hemoglobin level increased from 10.0 to 11.0 g/dl (p < 0.05) . These results indicate that even a dialysate containing 70 EU/l of ET level may induce a chronic inflammatory state . hs-CRP is a very useful marker of chronic inflammation and the use of ultrapure dialysate is necessary to improve a chronic inflammatory state . The targeted ET level at the end of the dialysate should be set at </=1.0 EU/l . Copyright (c) 2004 S . Karger AG, Basel. Acc Chem Res, 2005 Jan, 38(1), 62 - 9 How to hide zinc in a small protein; Blindauer CA et al.; Small cysteine-rich proteins (metallothioneins) and related domains of some large proteins (e.g., lysine methyltransferases) bind tri- and tetranuclear zinc clusters with topologies resembling fragments of Zn(II) sulfide minerals . These clusters are ubiquitous in animals, plants, and bacteria . Bacterial metallothioneins can also contain histidines as cluster ligands and embed Zn(II) with a "treble-clef"-like finger fold . This unusual embedded Zn(II) is "hidden" and surprisingly inert toward Zn or Cd exchange . Clearly, proteins can exert fine control over both the thermodynamics and kinetics of zinc binding in thiolate clusters . Genome sequences suggest that related zinc-finger sites are common in a variety of bacteria. Int J Syst Evol Microbiol, 2005 Jan, 55(Pt 1), 281 - 7 Methylobacterium hispanicum sp . nov . and Methylobacterium aquaticum sp . nov., isolated from drinking water; Gallego V et al.; Members of the genus Methylobacterium are ubiquitous in nature and can be isolated from almost any freshwater environment where dissolved oxygen exists . This genus is composed of a variety of pink-pigmented, facultatively methylotrophic (PPFM) bacteria . During a screening programme to monitor the bacterial population present in the drinking water of a municipal water supply in Seville (Spain) during the year 2003, five strains of PPFM bacteria were isolated and characterized . Analysis of their complete 16S rRNA gene sequences revealed that they constituted two separate phylogenetic groups (strains GP34(T) and GR18, and strains GR16(T), GP22 and GP32, respectively) showing highest similarity to members of the genus Methylobacterium . The highest 16S rRNA sequence similarities of strain GP34(T) were found with respect to the type strains of Methylobacterium radiotolerans (96.6 %) and Methylobacterium fujisawaense (96.4 %) and the highest 16S rRNA sequence similarities of strain GR16(T) were to the type strains of Methylobacterium extorquens (96.0 %) and Methylobacterium rhodesianum (95.8 %) . The G+C content of their DNA ranged from 66.5 to 67.8 mol% . DNA-DNA hybridization studies confirmed that they constituted two separate genospecies . On the basis of this phenotypic, phylogenetic and genotypic study, two novel species of the genus Methylobacterium are proposed: Methylobacterium hispanicum sp . nov., with type strain GP34(T) (CECT 5997(T)=CCM 7219(T)=DSM 16372(T)=CIP 108332(T)), and Methylobacterium aquaticum sp . nov., with type strain GR16(T) (CECT 5998(T)=CCM 7218(T)=DSM 16371(T)=CIP 108333(T)). Plant Cell Physiol, 2004 Dec, 45(12), 1768 - 78 Interaction of N-Acetylglutamate Kinase with a PII-Like Protein in Rice; Sugiyama K et al.; PII protein in bacteria is a sensor for 2-oxoglutarate and a transmitter for glutamine signaling . We identified an OsGlnB gene that encoded a bacterial PII-like protein in rice . Yeast two-hybrid analysis showed that an OsGlnB gene product interacted with N-acetylglutamate kinase 1 (OsNAGK1) and PII-like protein (OsGlnB) itself in rice . In cyanobacteria, NAGK is a key enzyme in arginine biosynthesis . Transient expression of OsGlnB cDNA or OsNAGK1 cDNA fused with sGFP in rice leaf blades strongly suggested that the PII-like protein as well as OsNAGK1 protein is located in chloroplasts . Both OsGlnB and OsNAGK1 genes were expressed in roots, leaf blades, leaf sheaths and spikelets of rice, and these two genes were coordinately expressed in leaf blades during the life span . Thus, PII-like protein in rice plants is potentially able to interact with OsNAGK1 protein in vivo . This finding will provide a clue to the precise physiological function of PII-like protein in rice. Proc Natl Acad Sci U S A . 2005 Jan 14; {Epub ahead of print} Characterization of potential selenium-binding proteins in the selenophosphate synthetase system; Ogasawara Y et al.; Selenophosphate, an activated form of selenium that can serve as a selenium donor, is generated by the selD gene product, selenophosphate synthetase (SPS) . Selenophosphate is required by several bacteria and by mammals for the specific synthesis of Secys-tRNA, the precursor of selenocysteine in selenoenzymes . Although free selenide can be used in vitro for synthesis of selenophosphate, the physiological system that donates selenium to SPS is incompletely characterized . To detect potential selenium-delivery proteins, two known sulfurtransferases and glyceraldehyde-3-phosphate dehydrogenase (GAPDH; EC 1.2.1.12) were analyzed for ability to bind and transfer selenium . Rhodanese (EC 2.8.1.1) was shown to bind selenium tightly, with only part of the selenium being available as substrate for SPS in the presence of added reductant . 3-Mercaptopyruvate sulfurtransferase (3-MST; EC 2.8.1.2) and GAPDH also bound selenium supplied as selenodiglutathione formed from SeO3(2-) and glutathione . Selenium bound to 3-MST and GAPDH was released more readily than that from rhodanese and also was more available as a substrate for SPS . Although rhodanese retained tightly bound selenium under aerobic conditions, the protein gradually became insoluble, whereas GAPDH containing bound selenium was stable at neutral pH for a long period . These results indicate that 3-MST and GAPDH have more suitable potentials as a physiological selenium-delivery protein than rhodanese . In the presence of a selenium-binding protein, a low level of selenodiglutathione formed from SeO3(2-) and glutathione could effectively replace the high concentrations of selenide routinely used as substrate in the SPS in vitro assays. Circ Res . 2005 Jan 13; {Epub ahead of print} Nod1-Mediated Endothelial Cell Activation by Chlamydophila Pneumoniae; Opitz B et al.; Seroepidemiological and animal studies, as well as demonstration of viable bacteria in atherosclerotic plaques, have linked Chlamydophila pneumoniae infection to development of chronic vascular lesions and coronary heart disease . Inflammation and immune responses are dependent on host recognition of invading pathogens . The recently identified cytosolic Nod proteins are candidates for intracellular recognition of bacteria, such as the obligate intracellular chlamydia . In the present study, mechanisms of endothelial cell activation by C . pneumoniae via Nod proteins were examined . Viable, but not heat-inactivated, chlamydia activated human endothelial cells, suggesting that invasion of these cells is necessary for their profound activation . Endothelial cells express Nod1 . Nod1 gene silencing by small interfering RNA reduced C pneumoniae-induced IL-8 release markedly . Moreover, in HEK293 cells, overexpressed Nod1 or Nod2 amplified the capacity of C pneumoniae to induce nuclear factor kappaB (NF-kappaB) activation . Interestingly, heat-inactivated bacteria were still able to induced a NF-kappaB reporter gene activity via Nod proteins when transfected intracellularly, but not when provided from the extracellular side . In contrast, TLR2 sensed extracellular heat-inactivated chlamydia . In conclusion, we demonstrated that C pneumoniae induced a Nod1-mediated and Nod2-mediated NF-kappaB activation in HEK293 cells . In endothelial cells, Nod1 played a dominant role in triggering a chlamydia-mediated inflammatory process. Biol Chem, 2004 Dec, 385(12), 1185 - 92 Novel thioredoxin targets in Dictyostelium discoideum identified by two-hybrid analysis: interactions of thioredoxin with elongation factor 1alpha and yeast alcohol dehydrogenase; Brodegger T et al.; Thioredoxins (Trx) are ubiquitous dicysteine proteins capable of modulating enzymes and other cellular targets through specific disulfide-dithiol redox changes . They are unique in that a large number of very diverse metabolic systems are addressed and redox-regulated in bacteria, animal, and plant cells, but the finite number of thioredoxin interaction partners is still unknown . Two-hybrid methodology should provide a rational way to establish thioredoxin functions in a given organism . We report a search for physiological target proteins of thioredoxin1 in the social amoeba Dictyostelium discoideum , which possesses three developmentally regulated thioredoxin genes, all of which lack functional characterisation . A two-hybrid approach identified at least seven bona fide thioredoxin partners, including oxidoreductases, proteins of the ribosomal translation apparatus, and the cytoskeletal protein filopodin . With the exception of ribonucleotide reductase, none of these systems had previously been linked to specific redox modulation . Molecular interactions in two of the new thioredoxin/target protein couples were verified by biochemical studies: (1) thioredoxin1 and the abundant elongation factor 1alpha from D . discoideum form the mixed heterodisulfide characteristic of the thioredoxin mechanism of action; and (2) reduced thioredoxin, but not glutathione, strongly inhibits yeast alcohol dehydrogenase catalysis of ethanol oxidation. J Am Soc Mass Spectrom, 2005 Jan, 16(1), 46 - 59 Total structure characterization of unsaturated acidic phospholipids provided by vicinal di-hydroxylation of fatty acid double bonds and negative electrospray ionization mass spectrometry; Moe MK et al.; In the present work, the unsaturated fatty acid substituents of some phosphatidic acid, phosphatidylserine, phosphatidylinositol, and phosphatidylglycerol species were converted to their 1,2-di-hydroxy derivatives by OsO(4) . The subsequent electrospray ionization tandem low-energy mass spectrometry analysis of the deprotonated species allowed positional determination of the double bonds by the production of specific product-ions . The product-ions are formed by charge-remote and charge-proximate homolytic cleavages as well as charge-directed heterolytic cleavages and rearrangements . The commercial availability of pure species of the phospholipids in question was limited, and a number of species were therefore synthesized . The developed method was used to fully characterize the two isobaric phosphatidylglycerol species 16:0/16:1Delta(9) and 16:0/16:1Delta(10) extracted from the bacteria Methylococcus capsulatus . The presence of these fatty acids was supported by a gas-chromatography mass spectrometry investigation of the dimethyloxazoline derivatives of the species of the lipid extract . The present work demonstrates that a total structure characterization of acidic unsaturated phospholipids in isolate or in mixtures is accomplished by vicinal di-hydroxylation of olefinic sites and subsequent electrospray ionization mass spectrometry of the derivatized phospholipids. FEMS Microbiol Rev, 2005 Jan, 29(1), 65 - 81 The Icm/Dot type-IV secretion systems of Legionella pneumophila and Coxiella burnetii; Segal G et al.; Type-IV secretion systems are devices present in a wide range of bacteria (including bacterial pathogens) that deliver macromolecules (proteins and single-strand-DNA) across kingdom barriers (as well as between bacteria and into the surroundings) . The type-IV secretion systems were divided into two subgroups and Legionella pneumophila and Coxiella burnetii are the only two bacteria known today to utilize a type-IVB secretion system for pathogenesis . In this review we summarized the available information concerning the icm/dot type-IVB secretion systems by comparing the two bacteria that possess this system, the proteins components of their systems as well as the homology of proteins from type-IVB secretion systems to proteins from type-IVA secretion systems . In addition, the phenotypes associated with mutants in the L . pneumophila icm/dot genes, their relations to properties of specific Icm/Dot proteins as well as the protein substrates delivered by this system are described. J Chromatogr B Analyt Technol Biomed Life Sci, 2005 Feb 5, 815(1-2), 137 - 45 System, trends and perspectives of proteomics in dicot plants Part III: Unraveling the proteomes influenced by the environment, and at the levels of function and genetic relationships; Agrawal GK et al.; This review is devoted to the proteomics studies in dicotyledoneous (dicot) plants, such as Arabidopsis, Medicago, potato, soybean, and tomato, under the influence of the environment and at the functional and genetic relationship levels, where the two core technologies, two-dimensional gel electrophoresis (2-DGE) and mass spectrometry (MS) have been instrumental in unraveling the proteomes affected therein . Abiotic and biotic stress responses, including the affect of allergens, the symbiotic interaction between the members of the Leguminoseae family and genera of nitrogen fixing bacteria, phosphoproteomics, and proteomics in revealing the genetic relationships between species and genera have been the subject of many proteomics studies, and these are discussed in this review . In all, these studies have complemented and extended the studies of developmental proteomics {G.K . Agrawal, M . Yonekura, Y . Iwahashi, H . Iwahashi, R . Rakwal, J . Chromatogr . B (2004)}. Biochem Biophys Res Commun, 2005 Feb 25, 327(4), 1094 - 9 Recombinant viral capsids as an efficient vehicle of oligonucleotide delivery into cells; Cooper A et al.; Delivery of oligonucleotides (ON) into cells is a technical challenge . In this study, we utilized the capsid of the hepatitis B virus (HBV) to meet this goal . A single and short open reading frame of the virus programs efficient capsid production in bacteria . We show that these capsids can encapsulate ON in vitro and then mediate their delivery into cells with extreme efficiency . This process is cell type non-specific, rendering the recombinant HBV capsid a potentially valuable vehicle for ON delivery into a wide range of cultured cells. Microb Pathog, 2005 Jan, 38(1), 41 - 6 Ultrastructural analysis of the interactions between Bordetella pertussis, Bordetella parapertussis and Bordetella bronchiseptica and human tracheal epithelial cells; Gueirard P et al.; Bordetella pertussis, Bordetella parapertussis and Bordetella bronchiseptica are respiratory pathogens that colonize the respiratory tract of their host after adhesion to the respiratory epithelium . Presently, the intracellular fate of these bacteria in human tracheal epithelial cells was compared by use of transmission electron microscopy . The three species, even when cytotoxic, were taken-up by epithelial cells . Although, some intracellular bacteria appeared morphologically intact and survived a few days inside epithelial cells, most of them appeared quickly degraded, phenomenon which was associated with an intense cell metabolic activity . Even cytotoxic Bordetella species is ultimately killed by human epithelial cells. Mol Cell Probes, 2005 Feb, 19(1), 41 - 50 Epub 2004 Nov 11. DNA microarray-based detection and identification of Chlamydia and Chlamydophila spp; Sachse K et al.; A microarray hybridization assay for identification of chlamydiae was developed using the ArrayTubetrade mark platform . The technology is comparatively inexpensive and involves plastic tube-integrated microchips and signal amplification by enzyme-catalyzed silver precipitation . Hybridization probes were designed on the basis of the most variable window approach, which identified species-specific nucleotide polymorphisms in a region of generally high sequence similarity . The selected 26-nt probe sequences were used on two different series of customized microarrays, i.e . combinatorial high-density in situ synthesized arrays and low-density spotted arrays . Target DNA was prepared by consensus PCR amplifying a 1-kbp segment of the ribosomal RNA operon . Unique species-specific hybridization patterns were obtained for all nine species of the family Chlamydiaceae on both microarray types . The present assay proved suitable for unambiguous species identification of chlamydial cell cultures and showed a potential for direct detection of these bacteria from clinical tissue. J Calif Dent Assoc, 2004 Nov, 32(11), 901 - 6 Airborne spread of disease--the implications for dentistry; Harrel SK; The potential for the airborne spread of disease has been recognized for many years . Recent studies have shown that this mode for disease transmission is capable of spreading a fatal disease such as Severe Acute Respiratory Syndrome over a wide area . Many dental procedures produce extensive aerosols and splatter that are routinely contaminated with bacteria, viruses, and blood . In the past, the potential for these aerosols and splatter to be a vector for disease spread has not been emphasized in dental infection control . Recently published data shows a need to reassess the potential for dental aerosols and splatter to spread disease and the need for their control . Simple and inexpensive methods for the control of dental aerosols and splatter are given . Dental personnel are urged to make the control of aerosols a standard part of their infection control procedures. Nihon Kokyuki Gakkai Zasshi, 2004 Nov, 42(11), 970 - 4 {Intense 18F-fluorodeoxyglucose uptake caused by pulmonary Mycobacterium intracellulare infection}; Matsuyama W et al.; A 70-year-old woman was admitted to our hospital for the evaluation of a pulmonary nodule in the left S5 segment . On 18F-fluorodeoxyglucose positron emission tomography (18FDG-PET), the nodule showed substantial uptake of 18F-fluorodeoxyglucose . Bronchoscopy was performed, but the cytology was negative . For a pathological diagnosis, a lung biopsy was carried out using video-associated thoracoscopy . The biopsy specimen showed granuloma formation with multinuclear giant cells . An acid-fast bacteria culture of the specimen was positive for Mycobacterium intracellulare . An atypical mycobacterium infection should be considered as a possibility when the 18FDG-PET of patients with pulmonary nodules is interpreted. Bundesgesundheitsblatt Gesundheitsforschung Gesundheitsschutz, 2005 Jan, 48(1), 76 - 83 {Cosmetic colorants . Toxicology and regulation.}; Platzek T et al.; Some recent publications raised concern over a possible link between hair dye use and the incidence of bladder tumours in a Californian population . The Scientific Committee for Cosmetic Products and Non-Food Products intended for Consumers (SCCNFP) demanded the toxicological testing of all hair dyes used in Europe to exclude any risk . The EU commission initiated corresponding measures . Only safe hair dyes will be included on a positive list while all other hair dyes will be banned . The hair dye lawsone-the dyeing ingredient of henna-was evaluated by the SCCNFP as genotoxic but the BfR came to another conclusion . The regulation of both lawsone and henna remains an open question . Furthermore, some cosmetic colorants were critically discussed . The azo dyes CI 12150, CI 26100, CI 27290 and CI 20170 are allowed for use in cosmetics . On cleavage they form the carcinogenic aromatic amines o-anisidine, 4-aminoazobenzene and 2,4-xylidine, respectively . For three of these dyes the cleavage by human skin bacteria in vitro to the respective arylamine was shown experimentally . Further problems may arise from colorants used for tattoos and permanent makeup . These products up to now are not subject to legislation and there are no regulatory stipulations with respect to health safety and purity for colorants used for these purposes. Methods, 2005 Feb, 35(2), 158 - 64 Epub 2004 Dec 20. In vivo stress preconditioning; Pespeni M et al.; The heat shock or stress protein response is a highly conserved defense mechanism . Activation of the stress protein response by mild hyperthermia or by pharmacological agents allows cells to withstand a subsequent metabolic insult that would otherwise be lethal, a phenomenon referred as "thermotolerance" or "preconditioning." Heat shock response is characterized by increased expression of stress proteins that provide cellular protection, e.g., via increased chaperoning activity in all organisms, from bacteria to animals and humans . Indeed, there is experimental evidence that overexpression of specific heat shock proteins or heat shock factors produce protective effects similar to those observed after stress preconditioning . The purpose of this review is first to discuss the methods used to induce in vivo thermotolerance with mild hyperthermia or pharmacological agents . Then, as an example of the organ protection provided by in vivo stress preconditioning, the second part of this paper will examine how the induction of thermotolerance modulates the lung inflammatory response associated with acute lung injury, thus providing broad organ and tissue protection against oxidative stress associated this syndrome. Methods, 2005 Feb, 35(2), 117 - 25 Epub 2004 Dec 19. Measurement of protein stability and protein denaturation in cells using differential scanning calorimetry; Lepock JR; Many methods exist for measuring and studying protein denaturation in vitro . However, measuring protein denaturation in cells under conditions relevant to heat shock presents problems due to cellular complexity and high levels of light scattering that interfere with optical techniques . A general method for measuring protein denaturation in cells using high sensitivity differential scanning calorimetry (DSC) is given . Profiles of specific heat (c(p) vs . temperature) are obtained providing information about transitions in cellular components including the denaturation of proteins . The specific approaches employed with erythrocytes, bacteria, and mammalian cells are described, and an identification of several features of the DSC profiles is given . Protein denaturation on the level of roughly 7-20% occurs for commonly used heat shocks in mammalian cells. Comp Biochem Physiol B Biochem Mol Biol, 2005 Feb, 140(2), 251 - 7 Digestive capabilities reflect the major food sources in three species of talitrid amphipods; Johnston M et al.; Digestive enzyme activities of three talitrid amphipods were examined to investigate the relationship between their digestive capabilities and diet . Laminarinase, cellobiase, carboxymethyl-cellulase, xylanase, alpha- and beta-glucosidase and lipase were detected in all three species suggesting talitrid amphipods can readily digest dietary carbohydrate and lipid, including complex polysaccharides . Relatively high specific enzyme activity (Units (mg(-1) digestive tract protein)(-1)) of laminarinase and lipase was detected in Talorchestia marmorata, a supralittoral kelp feeder which is coherent with the digestion of lipid-esters and beta-glucans (laminarin) which are the main lipid and storage polysaccharides of brown seaweeds . Talorchestia sp., a low shore intertidal feeder, had high enzymatic activity of alpha- and beta-glucosidase, cellobiase and xylanase, which is consistent with the digestion of diatoms . Keratroides vulgaris, a forest litter feeder had a relatively low specific activity of all enzymes . It is possible that leaf litter is partially digested prior to ingestion by bacteria and fungi present in the rotting vegetation, with bacterial and fungal enzymes contributing to this species' ability to hydrolyse its diet . This study provides the first quantitative data on digestive capacity in these three talitrid amphipods and confirms the relationship between dietary preference and digestive enzyme complement. Virus Res, 2005 Feb, 107(2), 117 - 27 The error threshold; Biebricher CK et al.; Basic principles underlying the population dynamics of bacteria and viruses are presented, with emphasis on RNA viruses . Concepts reviewed here include fitness, mutant generation, competition, selection, sequence space and the theoretical origins of quasispecies . A "wild-type" virus is no longer viewed as the fittest type, but as the center of gravity of a population landscape . A consequence of quasispecies is the existence of an error threshold for selective competence . The error threshold has a justification in quasispecies theory and lends itself to exploration of strategies to drive viral populations over the error threshold, the central theme of this volume. Biochem Biophys Res Commun, 2005 Feb 18, 327(3), 884 - 93 Evaluation of support matrices for immobilization of anaerobic consortia for efficient carbon cycling in waste regeneration; Chauhan A et al.; Efficient metabolism of fatty acids during anaerobic waste digestion requires development of consortia that include "fatty acid consuming H(2) producing bacteria" and methanogenic bacteria . The objective of this research was to optimize methanogenesis from fatty acids by evaluating a variety of support matrices for use in maintaining efficient syntrophic-methanogenic consortia . Tested matrices included clays (montmorillonite and bentonite), glass beads (106 and 425-600mum), microcarriers (cytopore, cytodex, cytoline, and cultispher; conventionally employed for cultivation of mammalian cell lines), BioSep beads (powdered activated carbon), and membranes (hydrophilic; nylon, polysulfone, and hydrophobic; teflon, polypropylene) . Data obtained from headspace methane (CH(4)) analyses as an indicator of anaerobic carbon cycling efficiency indicated that material surface properties were important in maintenance and functioning of the anaerobic consortia . Cytoline yielded significantly higher CH(4) than other matrices as early as in the first week of incubation . 16S rRNA gene sequence analysis from crushed cytoline matrix showed the presence of Syntrophomonas spp . (butyrate oxidizing syntrophs) and Syntrophobacter spp . (propionate oxidizing syntrophs), with Methanosaeta spp . (acetate utilizing methanogen), and Methanospirillum spp . (hydrogen utilizing methanogen) cells . It is likely that the more hydrophobic surfaces provided a suitable surface for adherence of cells of syntrophic-methanogenic consortia . Cytoline also appeared to protect entrapped consortia from air, resulting in rapid methanogenesis after aerial exposure . Our study suggests that support matrices can be used in anaerobic digestors, pre-seeded with immobilized or entrapped consortia on support matrices, and may be of value as inoculant-adsorbents to rapidly initiate or recover proper system functioning following perturbation. J Eur Acad Dermatol Venereol, 2005 Jan, 19(1), 107 - 111 Persistent unilateral orbital and eyelid oedema as a manifestation of Melkersson-Rosenthal syndrome; Cocuroccia B et al.; ABSTRACT Melkersson-Rosenthal syndrome (MRS) is a complex neuromucocutaneous disorder characterized by localized orofacial oedema and cranial nerve dysfunction, frequently associated with minor signs, including furrowed tongue . Complete forms are rare whereas mono- and oligosymptomatic variants are more common . A 71-year-old man presented with a 2-year history of relapsing and progressively persistent oedema of the right eyelids and periorbital region . A fissured tongue and telangiectatic rosacea had been present since the age of 50 and 60 years, respectively . The patient was also affected by essential hypertension and diabetes mellitus . A skin biopsy showed a marked upper dermal oedema, and small epithelioid cell granulomas arranged in perivascular and perilymphatic location . Collections of small epithelioid cells were occasionally observed within lymphatic spaces . No acid-fast bacteria, fungi or foreign bodies were detected . Intralesional corticosteroids induced transient improvement, whereas minocycline, clofazimine and dapsone have been ineffective . MRS may present with unilateral eyelid and periorbital swelling . Differential diagnoses of such cases may include a variety of cutaneous, ophthalmic and systemic diseases. Dis Aquat Organ, 2004 Nov 23, 62(1-2), 75 - 83 Species-specific polymerase chain reaction primer sets for the diagnosis of Tenacibaculum maritimum infection; Avendano-Herrera R et al.; In this study the specificity and sensitivity of 2 primer pairs, MAR1-MAR2 and Mar1-Mar2, for the detection of Tenacibaculum maritimum were evaluated in parallel using 79 T . maritimum strains isolated from different fish species, as well as 53 representatives of related and unrelated bacterial species . Both primer pairs were species-specific for T . maritimum, since no amplification products were obtained from chromosomal DNA of the non-T . maritimum bacteria tested . However, whereas MAR1-MAR2 identified all the T . maritimum strains studied, producing a unique and clear PCR band of the expected 1088 bp length, the Marl-Mar2 primer pair failed to amplify the 400 bp specific band in 3 sole isolates . To verify if these strains belonged to T . maritimum species, 2 endonucleases (PvuI and SacII) were selected as the most adequate enzymes to confirm the specificity of the MAR1-MAR2 amplified fragment . The digestion patterns obtained with both endonucleases supported the assignation of all the strains to T . maritimum . The sensitivity of both PCR detection methods was also different, showing a reduction of sensitivity in at least one order of magnitude of the Marl-Mar2 primer pair in comparison with MAR1-MAR2 . When the MAR-MAR2 PCR protocol was applied to different seeded turbot tissues, the detection limit was 10(2) to 10(4) T . maritimum cells per reaction . In addition, a nested PCR protocol for detection of this pathogens based on MAR1-MAR2 was developed, which increased the sensitivity by approximately 2 orders of magnitude, ranging from 1 to 250 T . maritimum cells per reaction depending on the tissue employed . The tissues that allowed the most easy detection of T . maritimum were the skin and mucus . Based on the findings reported here, we propose the nested PCR protocol as the most adequate for an accurate detection of T . maritimum in diagnostic pathology as well as in epidemiological studies of gliding bacterial disease of marine fish. Parasitology, 2004 Dec, 129(Pt 6), 655 - 61 Structure and synthesis of nematode phosphorylcholine-containing glycoconjugates; Houston KM et al.; Infection with filarial nematodes produces a chronic, long-lasting illness with adult worms able to survive within human hosts for up to 15 years . A contributor to the longevity of these parasites is the presence of phosphorylcholine (PC) on components of the worms' molecular secretions (ES) . PC on ES modulates host immune responses towards an anti-inflammatory phenotype thereby generating an environment favourable for parasite survival . PC is attached to nematode ES via a covalent association with carbohydrate, which, although well-documented in bacteria and fungi, is absent from humans, making it an ideal target for the development of novel drugs . In order to produce such drugs it is first necessary to understand the structure and synthesis of nematode PC-glycans . ES-62 is the major PC-ES-product of Acanthocheilonema viteae and is a homologue of PC-ES found in human filarial nematodes . We have studied the structure and biosynthesis of PC-glycans of ES-62 by a combination of pulse-chase experiments, experiments involving the use of inhibitors of each of intracellular trafficking, oligosaccharide processing and phospholipid biosynthesis and various forms of mass spectrometry . Our indications indicate that PC is transferred in the lumen of the medial Golgi to an N-type glycan consisting of a trimannosyl core with or without core fucosylation bearing between 1 and 4 N-acetyl glucosamine residues . The structure of the PC-N-glycans found in ES-62 appears to be conserved amongst filarial nematodes in that it has additionally been identified in Onchocerca volvulus and O . gibsoni . Also, similar structures have been found in non-filarial parasitic nematodes and in the free-living nematode Caenorhabditis elegans . Finally, PC has also been recently found attached to the carbohydrate moieties of nematode glycosphingolipids and the structure of these will also be considered. Dev Biol (Basel), 2000, 102, 149 - 55 Photochemical decontamination of red blood cell concentrates with the silicon phthalocyanine PC 4 and red light; Ben-Hur E et al.; Various approaches are being developed for virus inactivation of red blood cell concentrates (RBCC) in order to increase the safety of the blood supply . We have been studying the silicon phthalocyanine Pc 4 for this purpose, a photosensitizer activated with red light . Pc 4 targets the envelope of pathogenic viruses such as HIV . To protect RBC during the process two main approaches are used: (i) inclusion of quenchers of reactive oxygen species produced during the treatment . Tocopherol succinate was found to be most effective for this purpose; (ii) formulation of Pc 4, a lipophilic compound, in liposomes that reduce its binding to RBC but not to viruses . As a light source we used a light emitting diode array emitting at 670-680 nm . An efficient mixing device ensures homogenous light exposure during treatment of intact RBCC . Treatment of 50 ml RBCC with 5 microM Pc 4 and 18 J/cm2 light results in the inactivation of > or = 5.5 log10 HIV, > or = 6.3 log10 VSV and > or = 5 log10 of PRV and BVDV . The relative sensitivities of these viruses based on the slope of virus kill versus light dose are 1.0, 1.25, 1.5 and 1.9 for HIV, VSV, PRV and BVDV, respectively . To achieve the same level of virus inactivation in 350 ml RBCC, the light dose needed is 40 J/cm2 . HIV actively replicating in CEM cells is as sensitive as cell-free and HIV in latently infected cells is 3-4 times more sensitive . Parasites that can be transmitted by blood transfusion (P . falciparum and T . cruzi) are even more sensitive than viruses . Following treatment, RBCC can be stored for 28 days at 4 degrees C with haemolysis below 1% . Previous studies under less favourable conditions showed that baboon RBC circulated with an acceptable 24 hr recovery and half-life . Genetic toxicological studies of Pc 4 with or without light exposure (mutagenicity in bacteria, mammalian cells in vitro and clastogenicity in vivo) were negative . We conclude that a process using Pc 4 and red light can potentially reduce the risk of transmitting pathogens in RBCC. Riv Biol, 2004 Jan-Apr, 97(1), 79 - 90 The possible origin of the first cell biosystem in the thermal subsurface environment of the earth; Trevors JT; Bacteria are the simplest living biosystems or organisms that exhibit all the characteristics of life . As such, they are excellent models to examine the cell as the basic unit of life and the cell theory which states that all organisms are composed of one or more similar cells . In this article I examine the hypothesis that the primordial soup so often referred to in science was possibly an oil/water interface and/or emulsion in the Earth's, warm, anaerobic subsurface . This warm subsurface location, protected from surface radiation, could have been a favourable location for the assembly of the first bacterial cells on the Earth capable of growth and controlled division or the first biosystem. Curr Top Microbiol Immunol, 2005, 288, 1 - 8 Introduction and history of foot-and-mouth disease virus; Mahy BW; Foot-and-mouth disease (FMD) has been recognized as a significant epidemic disease threatening the cattle industry since the sixteenth century, and in the late nineteenth century it was shown by Loeffler and Frosch to be caused by a submicroscopic, filterable transmissible agent, smaller than any known bacteria . The agent causing FMD was thus the first virus of vertebrates to be discovered, soon after the discovery of tobacco mosaic virus of plants . It was not until 1920 that a convenient animal model for the study of FMD virus was established by Waldmann and Pape, using guinea-pigs, and with the later development of in vitro cell culture systems for the virus, the chemical and physical properties of FMD virus were elucidated during the remainder of the twentieth century, culminating in 1989 with a complete description of the three-dimensional structure of the virion . FMD virus is classified as a species in the Aphthovirus genus of the family Picornaviridae . The virus is acid labile, and the genome RNA contains a characteristic tract of polyC located about 360 nucleotides from the 5' terminus . Seven main serotypes exist throughout the world, as well as numerous subtypes . The World Reference Laboratory for FMD is located at Pirbright, Surrey, UK and undertakes surveillance of FMD epidemics by serotyping as well as by genotyping isolates of the virus . A major epidemic of FMD occurred in the UK in 2001 and was caused by a virulent strain of FMD virus with origins in Asia . The advantages and some disadvantages of controlling FMD outbreaks by vaccination are discussed. J Med Virol, 2005 Mar, 75(3), 470 - 4 Rapid and sensitive detection of mumps virus RNA directly from clinical samples by real-time PCR; Uchida K et al.; A rapid, sensitive, and specific assay to detect mumps virus RNA directly from clinical specimens using a real-time PCR assay was developed . The assay was capable of detecting five copies of standard plasmid containing cDNA from the mumps virus F gene . No cross-reactions were observed with other members of Paramyxoviridae, or with viruses or bacteria known to be meningitis pathogens . Seventy-three clinical samples consisting of throat swabs collected from patients with parotitis, and cerebrospinal fluid (CSF) collected from patients with aseptic meningitis, were examined with a real-time PCR assay developed by the authors, reverse-transcription nested-PCR (RT-n-PCR), and virus isolation using cell culture . Like the RT-n-PCR assay, the real-time PCR assay could detect mumps virus RNA in approximately 70% of both throat swabs and CSF samples, while, by tissue culture, mumps virus was isolated from only approximately 20% of CSF and 50% of throat swab samples . In addition, the real-time PCR assay could be developed easily into a quantitative assay for clinical specimens containing more than 1,800 copies of mumps virus RNA/ml by using serial dilutions of the standard plasmid . The results suggest that the real-time PCR assay is useful for identification of mumps virus infections, not only in typical cases, but also in suspected cases, which show only symptoms of meningitis or encephalitis . J . Med . Virol . 75:470-474, 2005 . (c) 2005 Wiley-Liss, Inc. Ann Rheum Dis, 2005 Feb, 64(2), 235 - 8 No evidence for involvement of the Toll-like receptor 4 (TLR4) A896G and CD14-C260T polymorphisms in susceptibility to ankylosing spondylitis; van der Paardt M et al.; OBJECTIVES: Ankylosing spondylitis (AS) is a multifactorial and polygenic disease . Apart from HLA, other genes very probably play a role in disease susceptibility . Indigenous bacteria of the gastrointestinal flora appear to play a role in the pathogenesis of the disease; therefore, genes controling the innate and acquired immune response are good candidates to study disease susceptibility . CD14 and Toll-like receptor 4 (TLR4) are key receptors for the sensing of bacteria . The CD14 C-260T and TLR4 A896G single nucleotide polymorphims are associated with aberrant signal transduction for bacterial agonists . METHODS: The distribution of the CD14 C-260T and TLR4 A896G polymorphisms was studied in genomic DNA from 113 unrelated white Dutch AS patients and 170 ethnically matched healthy controls . The diagnosis of AS was made according to the modified New York criteria . The CD14 C-260T and TLR4 A896G polymorphisms were genotyped by PCR-RFLP methods . RESULTS: No significant differences were found between patients and controls in the frequencies of the carriership of the less frequent CD14-260T allele (odds ratio 0.65; 95% confidence interval 0.37 to 1.15) or the TLR4 896G allele (1.68; 0.67 to 4.19) . CONCLUSIONS: There is no evidence for involvement of the CD14 C-260T or TLR4 A896G polymorphisms in susceptibility to AS . An important role of bacteria and genetic predisposition of the innate immune system in cases of AS cannot be excluded by these findings . Therefore, studies of the surprisingly highly polymorphic candidate genes in this field should be continued. New Microbiol, 2004 Oct, 27(4), 345 - 51 Prevalence of Mycoplasma pneumoniae and Chlamydia pneumoniae in pneumonia patients in four major hospitals in Trinidad; Nagalingam NA et al.; The prevalence of current Mycoplasma pneumoniae and Chlamydia pneumoniae infections in patients with pneumonia in Trinidad, and the relationship between pneumonia and risk factors were investigated . Blood samples were collected from 132 patients diagnosed by attending physicians, as suffering from pneumonia at four hospitals in Trinidad . Serum samples were tested for M . pneumoniae IgM and IgG and C . pneumoniae IgM by the enzyme immunoassay (EIA) . In addition, C . pneumoniae IgM and IgG were detected using microimmunofluorescence (MIF) . A comprehensive questionnaire which addressed demographic information as well as risk factors for pneumonia was administered to patients . All analyses were done using the Statistical Package for Social Sciences (SPSS), version 9 . Seroprevalences of 46.0% (58 of 126) were found for C . pneumoniae Ig M/G, and 66.7% (88 of 132) for M . pneumoniae Ig M/G . The difference was statistically significant (p < 0.01; chi2) . Thirty-four percent (43 of 125) for C . pneumoniae Ig M/acute Ig G and 28.8% (36 of 125) of M . pneumoniae IgM were not statistically significant (p > 0.05; chi2) . Hospital, gender and ethnicity of patients did not significantly (p > 0.05; chi2) affect the seroprevalence of the bacteria assayed for . However, the prevalence of C . pneumoniae (23.3%) in patients under 21 years old compared to other age groups was statistically significant (p = 0.043; chi2) . Overall, the seroprevalence to both pathogens was not significantly (p > 0.05; chi2) affected by comorbidities and signs/symptoms . It was concluded that new infections by C . pneumoniae in pneumonia patients may be an important aetiological agent for the condition in Trinidad. Compend Contin Educ Dent, 2004 Sep, 25(9 Suppl 1), 9 - 13 Amorphous calcium phosphates for tooth mineralization; Tung MS et al.; The destruction of tooth structure through caries and erosive processes is due to two types of acidic challenges that affect the tooth in different ways . Acidic attack by cariogenic bacteria initially produces subsurface lesions that weaken the enamel and, if left unchecked, can progress through the enamel and dentin and eventually into the pulpal cavity . Erosive attack by acidic foods and beverages removes mineral from the surface of enamel and initially causes dulling and loss of tooth luster; if left unchecked, it can progress to a more severe loss of enamel thickness and contour . This article focuses on the potential means of improving the cosmetic appearance of teeth by depositing mineral into surface defects . Several approaches use the unique properties of amorphous calcium phosphate (ACP) compounds, which have the highest rates of formation and dissolution among all the calcium phosphates . ACP has been shown to rapidly hydrolyze to form apatite, similar to carbonated apatite, the tooth mineral . Products containing ACP or ingredients that form ACP can include toothpastes, mouth rinses, artificial saliva, chewing gums, topically applied coatings, and other vehicles for topical use . When applied, they readily precipitate ACPs on and into tooth-surface defects . These products hopefully will provide users with new tools to restore and enhance the smoothness and luster of their teeth. Compend Contin Educ Dent, 2004 Jul, 25(7 Suppl 1), 54 - 7 Rationale for the daily use of a dentifrice containing triclosan in the maintenance of oral health; DeVizio W et al.; This paper provides an overview and summary of the data that demonstrate the effectiveness of a dentifrice containing triclosan/copolymer/fluoride in significantly reducing plaque, calculus, gingivitis, and the onset and progression of periodontitis . The caries-preventive benefit of the triclosan/copolymer/fluoride dentifrice was at least as good as that of a fluoride dentifrice . No evidence of bacterial resistance or the development of pathogenic or opportunistic bacteria was observed. Compend Contin Educ Dent, 2004 Jul, 25(7 Suppl 1), 38 - 45 Evidence that diabetes mellitus aggravates periodontal diseases and modifies the response to an oral pathogen in animal models; Graves DT et al.; Bacterial plaque has been shown to initiate periodontal diseases . Most studies indicate that the host response, rather than the direct effect of bacteria, is responsible for much of the destruction associated with periodontitis . Bacteria or their products have an indirect role by stimulating inflammation, which is associated with the excessive production of inflammatory mediators, such as prostaglandins, or cytokines, such as tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 . These mediators, in turn, induce the production and activation of enzymes that destroy gingival connective tissue and stimulate the formation of osteoclasts to resorb bone . Based on results in animal models and studies in humans showing that similar responses occur, the initial steps in the breakdown of connective tissue attachment to the tooth surface and bone resorption involve the production of inflammatory cytokines . Moreover, the risk and severity of periodontal diseases is affected by systemic factors, such as diabetes . Diabetes in particular seems to impair the ability to produce new bone formation after bone loss by preventing the formation of new bone that normally occurs after bone is resorbed, a process called coupling . In addition, the cytokines that stimulate loss of tissue, particularly TNF-alpha, may kill the cells that repair damaged connective tissue or bone . In diabetes there may be more TNF-alpha produced, leading to an even more limited capacity to repair tissue . The diminished capacity to form new bone may make it more difficult for diabetics in particular to repair the loss of tissue that occurs in periodontal diseases. Zhong Xi Yi Jie He Xue Bao, 2005 Jan, 3(1), 6 - 9 {Application and projects approved for fundamental research on integrated traditional Chinese and Western medicine in National Natural Science Foundation of China from 2000 to 2004.}; Wang CE et al.; From 2000 to 2004, the National Natural Science Foundation of China (NSFC) accepted 1 171 applications and funded 160 projects for fundamental research on integrated traditional Chinese and Western medicine . The success rate is 13.64% . Being supported by NSFC, a number of achievements well known in China and abroad have been made, such as acupuncture complementary anaesthesia, treatment of leukemia, viral hepatitis, cardio-cerebrovascular diseases, acute abdomen, burns and fracture with integrated traditional Chinese and Western medicine and their therapeutic mechanisms, and some new concepts and theories have been put forward, such as the theories of activating blood to resolve stagnation, simultaneous treatment of bacteria and toxin, etc . But there still exit some problems in the research of the funded projects . The research ways are mainly combination of different methods instead of integration of both traditional Chinese and Western medicine . The research results are mainly confirmation of traditional Chinese medical theories instead of exploration of their innate regularities . The relationship among disease, syndrome and symptom is not clearly explored . The principal-subordinate relationship between macro- and micro-differentiation of syndromes is not clear . Academic research is short of new ideas . Improper or biased explanation of the theories of traditional Chinese medicine still exists . On analysis of above problems, some preferable aspects of projects for fundamental research on integrated traditional Chinese and Western medicine funded by NSFC in future are suggested in this article. Mol Ecol, 2005 Jan, 14(1), 285 - 94 On the ubiquity and phylogeny of Wolbachia in lice; Kyei-Poku GK et al.; Abstract Wolbachia are intracellular bacteria that occur in an estimated 20% of arthropod species . They are of broad interest because they profoundly affect the reproductive fitness of diverse host taxa . Here we document the apparent ubiquity and diversity of Wolbachia in the insect orders Anoplura (sucking lice) and Mallophaga (chewing lice), by detecting single or multiple infections in each of 25 tested populations of lice, representing 19 species from 15 genera spanning eight taxonomic families . Phylogenetic analyses indicate a high diversity of Wolbachia in lice, as evidenced by the identification of 39 unique strains . Some of these strains are apparently unique to lice, whereas others are similar to strains that infect other insect taxa . Wolbachia are transmitted from infected females to their offspring via egg cyto-plasm, such that similar species of lice are predicted to have similar strains of Wolbachia . This predicted pattern is not supported in the current study and may reflect multiple events of recent horizontal transmission between host species . At present, there is no known mechanism that would allow for this latter mode of transmission to and within species of lice. Environ Microbiol, 2005 Jan, 7(1), 140 - 6 Variations in the abundance and identity of class II aromatic ring-hydroxylating dioxygenase genes in groundwater at an aromatic hydrocarbon-contaminated site; Taylor PM et al.; Summary The abundance of genes encoding aromatic ring-hydroxylating dioxygenases (RHDs) in the groundwater at an aromatic hydrocarbon-contaminated landfill near Sydney, Australia, was determined by quantitative DNA-DNA hybridization using class II RHD genes as probes . There were marked differences in hybridization signal intensity against DNA extracted from the groundwater at seven different locations across this heterogeneous site . This was interpreted as indicating variation in RHD gene abundance . Clone libraries of polymerase chain reaction (PCR)-amplified RHD gene fragments were constructed from DNA from each of the groundwater samples . The libraries from the samples with greater RHD gene abundance were dominated by a group of bacterial class II RHD genes, designated the S-cluster, that has yet to be found in cultured isolates . These groundwater samples contained no detectable petroleum hydrocarbons . A second group of class II RHD gene sequences, designated the T-cluster, dominated RHD gene clone libraries prepared from groundwater samples that contained detectable levels of total petroleum and aromatic hydrocarbons but lower RHD gene abundance . The hosts and in situ expression of these novel genes, and the substrates of the enzymes they encode, remain unknown . The scarcity of genes from known aromatic hydrocarbon-degrading bacteria and the numerical dominance of the novel genes suggest that the hosts of these novel genes may play an important role in aromatic hydrocarbon degradation at this site. Environ Microbiol, 2005 Jan, 7(1), 98 - 106 Environmental regulation of the anaerobic oxidation of methane: a comparison of ANME-I and ANME-II communities; Nauhaus K et al.; Summary The anaerobic oxidation of methane (AOM) is one of the major sinks for methane on earth and is known to be mediated by at least two phylogenetically different groups of anaerobic methanotrophic Archaea (ANME-I and ANME-II) . We present the first comparative in vitro study of the environmental regulation and physiology of these two methane-oxidizing communities, which occur naturally enriched in the anoxic Black Sea (ANME-I) and at Hydrate Ridge (ANME-II) . Both types of methanotrophic communities are associated with sulfate-reducing-bacteria (SRB) and oxidize methane anaerobically in a 1:1 ratio to sulfate reduction (SR) . They responded sensitively to elevated methane partial pressures with increased substrate turnover . The ANME-II-dominated community showed significantly higher cell-specific AOM rates . Besides sulfate, no other electron acceptor was used for AOM . The processes of AOM and SR could not be uncoupled by feeding the SRB with electron donors such as acetate, formate or molecular hydrogen . AOM was completely inhibited by the addition of bromoethanesulfonate in both communities, indicating the participation of methanogenic enzymes in the process . Temperature influenced the intensity of AOM, with ANME-II being more adapted to cold temperatures than ANME-I . The variation of other environmental parameters, such as sulfate concentration, pH and salinity, did not influence the activity of both communities . In conclusion, the ecological niches of methanotrophic Archaea seem to be mainly defined by the availability of methane and sulfate, but it remains open which additional factors lead to the dominance of ANME-I or -II in the environment. Environ Microbiol, 2005 Jan, 7(1), 47 - 54 Divergent transcriptional and translational signals in Archaea; Torarinsson E et al.; Summary Many Archaea, in contrast to bacteria, produce a high proportion of leaderless transcripts, show a wide variation in their consensus Shine-Dalgarno (S-D) sequences and frequently use GUG and UUG start codons . In order to understand the basis for these differences, 18 complete archaeal genomes were examined for sequence signals that are positionally conserved upstream from genes . These functional motifs include box A promoter sequences for leaderless transcripts and S-D sequences for transcripts with leaders . Most of the box A sequences were preceded by a BRE-like motif and followed by a previously undetected A/T peak centred on position -10 . Moreover, the sequence of the predominant S-D motifs in an archaeon is shown to depend on the precise number of nucleotides between the conserved anti-S-D CCUCC sequence and the 3'-terminal nucleotide of 16S RNA . Correlations with phylogenetic trees, constructed for the 18 Archaea, reveal that usage of high levels of both S-D motifs, and GUG and UUG start codons occurs exclusively in the shorter branched Archaea . High levels of leaderless transcripts are found in the longer branched Archaea. Trop Anim Health Prod, 2004 Nov, 36(8), 751 - 62 Effects of fresh cassava tops on rumen environment parameters, thyroid gland hormones and liver enzymes of local yellow cattle fed urea-treated fresh rice straw; Khang DN et al.; In a 4 x 4 Latin square design study, four rumen-cannulated local yellow cattle, average weight 330 kg, were fed urea-treated fresh rice straw (UFRS) ad libitum and given 1 kg cassava root meal daily as basal diet in each 30-day trial period . Four treatment levels of crude protein of fresh cassava tops (FCT) were supplied: 0 (FCT0), 50 (FCT50), 100 (FCT100) and 150 g (FCT150) crude protein per 100 kg body weight . The results showed a continuous decrease in dry matter intake (DMI) of UFRS with increasing level of FCT supplementation (p<0.001) . Total DMI for treatments FCT150, FCT100, FCT50 and FCT0 were 2.66, 2.42, 2.23 and 2.05 kg dry matter per 100 kg live weight per day, respectively . Increasing levels of FCT supplementation increased the concentration of total volatile fatty acids (p<0.05) and ammonia (p<0.001) and resulted in a decrease in pH (p<0.05) . Rumen protozoa population varied widely but was significantly lower with increasing FCT supplementation (p<0.001), while rumen bacteria population was higher (p<0.05) . Overall, average plasma triiodothyronine and thyroxine concentrations in the four groups were respectively 1.36, 0.94, 0.91, 0.80 ng/ml (p<0.01) and 60.9, 48.7, 45.2, 42.5 ng/ml (p<0.001) . There were non-significant differences in alanine aminotransferase and aspartate aminotransferase concentrations among the treatments. Proc Natl Acad Sci U S A, 2005 Jan 18, 102(3), 856 - 61 Epub 2005 Jan 10. A bipartite signal mediates the transfer of type IV secretion substrates of Bartonella henselae into human cells; Schulein R et al.; Bacterial type IV secretion (T4S) systems mediate the transfer of macromolecular substrates into various target cells, e.g., the conjugative transfer of DNA into bacteria or the transfer of virulence proteins into eukaryotic host cells . The T4S apparatus VirB of the vascular tumor-inducing pathogen Bartonella henselae causes subversion of human endothelial cell (HEC) function . Here we report the identification of multiple protein substrates of VirB, which, upon translocation into HEC, mediate all known VirB-dependent cellular changes . These Bartonella-translocated effector proteins (Beps) A-G are encoded together with the VirB system and the T4S coupling protein VirD4 on a Bartonella-specific pathogenicity island . The Beps display a modular architecture, suggesting an evolution by extensive domain duplication and reshuffling . The C terminus of each Bep harbors at least one copy of the Bep-intracellular delivery domain and a short positively charged tail sequence . This biparte C terminus constitutes a transfer signal that is sufficient to mediate VirB/VirD4-dependent intracellular delivery of reporter protein fusions . The Bep-intracellular delivery domain is also present in conjugative relaxases of bacterial conjugation systems . We exemplarily show that the C terminus of such a conjugative relaxase mediates protein transfer through the Bartonella henselae VirB/VirD4 system into HEC . Conjugative relaxases may thus represent the evolutionary origin of the here defined T4S signal for protein transfer into human cells. FEBS Lett, 2005 Jan 17, 579(2), 379 - 384 Efavirenz enhances the proteolytic processing of an HIV-1 pol polyprotein precursor and reverse transcriptase homodimer formation; Tachedjian G et al.; The non-nucleoside reverse transcriptase inhibitor, efavirenz (EFV), is a potent enhancer of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) p66/p51 heterodimerization . While the mechanism of RT heterodimer formation in HIV-1 infected cells is not completely understood, it has been speculated that Gag-Pol/Gag-Pol and/or RT homodimer interactions may represent important intermediates in the pathway . To elucidate whether EFV impacts on these interactions, we have evaluated the effects of this drug on RT homodimer interactions and HIV-1 Gag-Pol processing . EFV, but not nevirapine, significantly enhanced RT p66/p66 and p51/p51 homodimer interactions and accelerated the proteolytic cleavage of a model HIV-1 Pol polyprotein precursor expressed in bacteria . These data suggest that potent mediators of RT dimerization might interfere with the late-stages of viral replication. Brief Funct Genomic Proteomic, 2004 Nov, 3(3), 199 - 211 Operons in eukaryotes; Blumenthal T; It was thought that polycistronic transcription is a characteristic of bacteria and archaea, where many of the genes are clustered in operons composed of two to more than ten genes . By contrast, the genes of eukaryotes are generally considered to be monocistronic, each with its own promoter at the 5' end and a transcription terminator at the 3' end; however, it has recently become clear that not all eukaryotic genes are transcribed monocistronically . Numerous instances of polycistronic transcription in eukaryotes, from protists to chordates, have been reported . These can be divided into two broad types . Dicistronic transcription units specify a messenger RNA (mRNA) encoding two separate genes that is transported to the cytoplasm and translated in that form . Presumably, internal ribosome entry sites (IRES), or some form of translational re-initiation following the stop codon, are responsible for allowing translation of the downstream gene . In the other type, the initial transcript is processed by 3' end cleavage and trans-splicing to create monocistronic mRNAs that are transported to the cytoplasm and translated . Like bacterial operons, eukaryotic operons often result in co-expression of functionally related proteins. Arthritis Res Ther, 2005, 7(1), 1 - 11 Epub 2004 Nov 02. Mast cells in inflammatory arthritis; Nigrovic PA et al.; Mast cells are present in limited numbers in normal human synovium, but in rheumatoid arthritis and other inflammatory joint diseases this population can expand to constitute 5% or more of all synovial cells . Recent investigations in a murine model have demonstrated that mast cells can have a critical role in the generation of inflammation within the joint . This finding highlights the results of more than 20 years of research indicating that mast cells are frequent participants in non-allergic immune responses as well as in allergy . Equipped with a diversity of surface receptors and effector capabilities, mast cells are sentinels of the immune system, detecting and delivering a first response to invading bacteria and other insults . Accumulating within inflamed tissues, mast cells produce cytokines and other mediators that may contribute vitally to ongoing inflammation . Here we review some of the non-allergic functions of mast cells and focus on the potential role of these cells in murine and human inflammatory arthritis. J Clin Periodontol, 2005 Jan, 32(1), 45 - 52 Periodontitis, a marker of risk in pregnancy for preterm birth; Dortbudak O et al.; Dortbudak O, Eberhardt R, Ulm M, Persson GR: Periodontitis, a marker of risk in pregnancy for preterm birth . J Clin Periodontol 2004; doi: 10.1111/j.1600-051X.2004.00630.x . (c) Blackwell Munksgaard, 2004 . Abstract Background: Why chronic periodontitis may induce an inflammatory response with premature pregnancy termination is unclear . Aims: (1) To assess if periodontitis predicts premature gestation; (2) to study amniotic fluid cytokines and periodontitis variables in early-stage pregnancy . Material and Methods: A periodontal examination and collection of amniotic fluid was performed (weeks 15-20) of pregnancy in 36 women at risk for pregnancy complications . Amniotic fluid (bacteria), vaginal smears and intra-oral plaque samples were studied . Cytokine levels in amniotic fluid were studied in relation to other study variables . Results: Periodontitis was diagnosed in 20% of normal and in 83% of preterm birth cases (p<0.01) . Bacteria were never found in the amniotic fluids studied . Sub-gingival plaque samples including bacteria in the orange and red complexes were found in 18% of full-term 100% of preterm cases (p<0.001) and total colony-forming units (CFUs) were higher in preterm birth (p<0.01) . Amniotic levels of interleukin (IL)-6 and prostaglandin-E(2) (PGE(2)) were higher in preterm cases (p<0.001) . Amniotic IL-6 (r=0.56, p<0.01) and PGE(2) (r=0.50, p<0.01) cytokine levels were correlated with CFU from sub-gingival plaque samples (r(2)=0.44).The odds ratio of preterm delivery and having periodontitis was 20.0 (95% confidence interval (CI): 2.0-201.7, p<0.01) . The odds of >60 CFU in sub-gingival plaque and preterm birth was 32.5:1 (95% CI: 3.0-335.1, p<01) . Conclusions: Pregnant women with findings of elevated amniotic fluid levels of PGE(2), IL-6 and IL-8 in the 15-20 weeks of pregnancy and with periodontitis are at high risk for premature birth . The implication of this is that periodontitis can induce a primary host response in the chorioamnion leading to preterm birth. Biochemistry, 2005 Jan 18, 44(2), 555 - 64 Baboon Lipoprotein(a) Binds Very Weakly to Lysine-Agarose and Fibrin Despite the Presence of a Strong Lysine-Binding Site in Apolipoprotein(a) Kringle IV Type 10; Belczewski AR et al.; Human apolipoprotein(a) kringle IV type 10 {apo(a) KIV(10)} contains a strong lysine-binding site (LBS) that mediates the interaction of Lp(a) with biological substrates such as fibrin . Mutations in the KIV(10) LBS have been reported in both the rhesus monkey and chimpanzee, and have been proposed to explain the lack of ability of the corresponding Lp(a) species to bind to lysine and fibrin . To further the comparative analyses with other primate species, we sequenced a segment of baboon liver apo(a) cDNA spanning KIV(9) through the protease domain . Like rhesus monkey apo(a), baboon apo(a) lacks a kringle V (KV)-like domain . Interestingly, we found that the baboon apo(a) KIV(10) sequence contains all of the canonical LBS residues . We sequenced the apo(a) KIV(10) sequence from an additional 10 unrelated baboons; 17 of 20 alleles encoded Trp at position 70, whereas only two alleles encoded Arg at this position and thus a defective LBS . Despite the apparent presence of a functional KIV(10) LBS in most of the baboons, none of the Lp(a) in the plasma of the corresponding baboons bound specifically to lysine-Sepharose (agarose) even upon partial purification . Moreover, baboon Lp(a) bound very poorly to plasmin-modified fibrinogen . Expression of baboon and human KIV(10) in bacteria allowed us to verify that these domains bind comparably to lysine and lysine analogues . We conclude that presentation of KIV(10) in the context of apo(a) lacking KV may interfere with the ability of KIV(10) to bind to substrates such as fibrin; this paradigm may also be present in other non-human primates. J Biol Chem . 2005 Jan 7; {Epub ahead of print} A dictyostelium mutant with reduced lysozyme levels compensates by increased phagocytic activity; Muller I et al.; Lysozymes are bacteria-degrading enzymes and play a major role in the immune defense of animals . In free-living protozoa, lysozyme-like proteins are involved in the digestion of phagocytosed bacteria . Here, we purified a protein with lysozyme activity from Dictyostelium amoebae, which constitutes the founding member a novel class of lysozymes . By tagging the protein with GFP or the myc epitope, a new type of lysozyme containing vesicle was identified that was devoid of other known lysosomal enzymes . The most highly expressed isoform, encoded by the alyA gene, was knocked out by homologous recombination . The mutant cells had greatly reduced enzymatic activity and grew inefficiently when bacteria were the sole food source . Over time, the mutant gained the ability to internalize bacteria more efficiently, so that the defect in digestion was compensated by increased uptake of food particles. Yi Chuan, 2003 Jan, 25(1), 40 - 4 {Compatible-solute-supported periplasmic expression of recombinant immunotoxins and the effects of glucose thereby.}; Ren YM et al.; Bacteria protect themselves against high external osmolality by uptake or synthesis of a limited number of so-called compatible solutes.Under such extreme conditions,combine factors can facilitate the foreign recombinant protein expression.In our experiment,we added additional NaCl and two compatible solutes (glycine betaine and sorbitol) into the medium.The recombinant immunotoxin BDI(scFv)-PE38 was expressed mainly in soluble form and can be purified by Ni2+-NTA agarose.We also found the time when the glucose was added into the medium greatly affected the expression level of the recombinant immunotoxin.This suggested that glucose played an important role in the expression system. Contraception, 2005 Jan, 71(1), 65 - 7 A scanning electron microscopic study of the contraceptive vaginal ring; Miller L et al.; OBJECTIVE: To compare by electron microscopy the surface of vaginal contraceptive rings before and after use . METHODS: A single ring was worn for 28 days by a volunteer with normal vaginal flora . This worn ring was then cut in half, with one half placed directly into fixative and the other half rinsed to remove visible mucus prior to being placed into the fixative . Another ring was taken directly from the package and placed in fixative . The surfaces of the three ring samples were studied by scanning electron microscopy at varying magnifications to ascertain if there was penetration of bacteria into the vinyl polymer or if the ring surface was altered with 1 month of use . RESULTS: The ring half placed directly into fixative with visible mucus had cellular debris on its surface but no evidence of material penetration . The other half of the ring worn for 28 days and rinsed before fixation had a similar surface appearance to the unused ring . CONCLUSIONS: The surface of the ethylene vinyl acetate contraceptive vaginal ring is remarkably uniform and after 1 month of use was without visible evidence of erosion, embedded bacteria or structural changes compared to an unused ring. Rocz Akad Med Bialymst, 2004, 49 Suppl 1, 239 - 41 Attempts to detect Helicobacter pylori in atherosclerotic plaques; Sulewska A et al.; Cardiovascular and cerebrovascular diseases are regarded to be the main causes of mortality in developed countries, atherosclerosis being at their pathological base . During the recent years, attention was paid to the role of bacterial infections, including Helicobacter pylori, in the process of atherogenesis and coronary heart disease development . The aim of the study was an evaluation of H . pylori presence--by means of PCR technique--in atherosclerotic changes, obtained by endarterectomy, performed during coronary artery bypass grafting (CABG) . In the analysed group of patients, the following risk factors were found: hyperlipidaemia, smoking, hypertension, obesity, diabetes mellitus, cardiac infarction . No DNA of the bacteria was traced in any of the patients. Rocz Akad Med Bialymst, 2004, 49 Suppl 1, 222 - 4 B (CD20+) lymphocytes in the antrum mucosa of children with Helicobacter pylori infection; Maciorkowska E et al.; The aim of the study was to estimate the expression of CD20+ lymphocytes in the antrum mucosa in children, infected with Helicobacter pylori and after bacteria eradication . Biopsy specimens of gastric mucosa were the specimens, collected from 59 H . pylori-positive patients (Group I), 29 patients after H . pylori infection (Group II) and 18 H . pylori-negative children (Group III) . The collected specimens were assessed for infection and inflammation and the expression of CD20+ lymphocytes was estimated, using mice monoclonal antibodies . The expression of CD20+ lymphocytes in the inflammatory infiltrate of the antrum mucosa correlated with the severity of gastritis, found in children with Helicobacter pylori infection and was the highest in comparison with the group of children after H . pylori eradication. Rocz Akad Med Bialymst, 2004, 49 Suppl 1, 161 - 3 Assessment of potassium and sodium ion concentrations in the vitreous humour of swine isolated eyeballs after organism death; Brzezinski PM et al.; Potassium and sodium ion concentrations were estimated by the flame photometry and potentiometry in the vitreous fluid of isolated porcine eyeballs at time of death and of eyeballs, stored at temperature of 6-8 degrees C during post-mortem intervals: 4, 28, 52, 75, 100, 124 and 148 hours . The increase of K+ concentration and decrease of Na+ concentration were proportional to the increasing post-mortem time intervals . The results of the potentiometric measurements of K+ and sodium ion concentrations were significantly lower, as compared to those after flame photometry . In all the vitreous fluid smears after 124 and 148 hours, gram (-) bacteria were found . Our results suggest that bacterial infection participates in the variability of K+ levels . The influence of bacterial infection on the margin of error for the K+ post-mortem test remains unanswered and needs further studies. Eur Rev Med Pharmacol Sci, 2004 Sep-Oct, 8(5), 199 - 203 Immunoglobulin M memory B cell decrease in inflammatory bowel disease; Di Sabatino A et al.; BACKGROUND & OBJECTIVES: Memory B cells represent 30-60% of the B cell pool and can be subdivided in IgM memory and switched memory . IgM memory B cells differ from switched because they express IgM and their frequency may vary from 20-50% of the total memory pool . Switched memory express IgG, IgA or IgE and lack surface expression of IgM and IgD . Switched memory B cells derive from the germinal centres, whereas IgM memory B cells, which require the spleen for their survival and/or generation, are involved in the immune response to encapsulated bacteria . Since infections are one of the most frequent comorbid conditions in inflammatory bowel disease, we aimed to verify whether IgM memory B cell pool was decreased in Crohn's disease and ulcerative colitis patients . PATIENTS & METHODS: Peripheral blood samples were obtained from 22 Crohn's disease patients, 20 ulcerative colitis patients, 22 healthy controls and 18 splenectomized patients . To analyse peripheral blood lymphocytes, flow cytometry was performed using anti-CD19, anti-CD22, anti-CD27, anti-IgM, anti-IgD and anti-CD38 monoclonal antibodies . RESULTS: Circulating IgM memory B cells were significantly lower in Crohn's disease (median 7.1%, range 1.8-20.7) and ulcerative colitis patients (median 8.1%, range 2.1-18.8) in comparison to control subjects (median 14.0%, range 6.8-31.1) . As expected, there was a highly significant difference in the proportion of IgM memory B cells between splenectomized patients (median 2.4%, range 0.9-6.9) and healthy controls . Crohn's disease patients with abscesses showed the lowest frequency of IgM memory B cells . DISCUSSION: Our findings show that peripheral IgM memory B cells are reduced in inflammatory bowel disease patients . Further studies are necessary to answer the question of whether high risk of infection (abscess development) is promoted by the reduction/depletion of IgM memory B-cell pool in inflammatory bowel disease. Compend Contin Educ Dent, 2004 Oct, 25(10 Suppl 1), 4 - 7 A novel oral hygiene system through integration of a sonic toothbrush and liquid toothpaste; Dudgeon DJ et al.; A novel system for home oral hygiene, the IntelliClean System from Sonicare and Crest is an integrated sonic toothbrush and liquid-toothpaste dispensing system . New research has established the ability of this system to reduce plaque, especially in interproximal regions, and to reduce gingivitis . The unique capability for the user to reapply liquid toothpaste during brushing (ie, re-dose) allows for a greater reduction in bacteria in the sulcus and an enhanced in-use experience that leads to improved compliance with brushing instruction, specifically longer brushing times . Through in vitro and in vivo studies, the system and its constituents have been shown to be safe for daily use. Proc Natl Acad Sci U S A, 2005 Jan 18, 102(3), 850 - 5 Epub 2005 Jan 06. Discovery of a free-living chlorophyll d-producing cyanobacterium with a hybrid proteobacterial/cyanobacterial small-subunit rRNA gene; Miller SR et al.; Chlorophyll d-producing cyanobacteria are a recently described group of phototrophic bacteria that is a major focus of photosynthesis research, previously known only from marine environments in symbiosis with eukaryotes . We have discovered a free-living member of this group from a eutrophic hypersaline lake . Phylogenetic analyses indicated these strains are closely related to each other but not to prochlorophyte cyanobacteria that also use an alternative form of chlorophyll as the major light-harvesting pigment . We have also demonstrated that these bacteria acquired a fragment of the small-subunit rRNA gene encoding a conserved hairpin in the bacterial ribosome from a proteobacterial donor at least 10 million years before the present . Thus, our most widely used phylogenetic marker can be a mosaic of sequence fragments with widely divergent evolutionary histories. Proc Natl Acad Sci U S A, 2005 Jan 18, 102(3), 838 - 43 Epub 2005 Jan 06. The genome of the heartwater agent Ehrlichia ruminantium contains multiple tandem repeats of actively variable copy number; Collins NE et al.; Heartwater, a tick-borne disease of domestic and wild ruminants, is caused by the intracellular rickettsia Ehrlichia ruminantium (previously known as Cowdria ruminantium) . It is a major constraint to livestock production throughout subSaharan Africa, and it threatens to invade the Americas, yet there is no immediate prospect of an effective vaccine . A shotgun genome sequencing project was undertaken in the expectation that access to the complete protein coding repertoire of the organism will facilitate the search for vaccine candidate genes . We report here the complete 1,516,355-bp sequence of the type strain, the stock derived from the South African Welgevonden isolate . Only 62% of the genome is predicted to be coding sequence, encoding 888 proteins and 41 stable RNA species . The most striking feature is the large number of tandemly repeated and duplicated sequences, some of continuously variable copy number, which contributes to the low proportion of coding sequence . These repeats have mediated numerous translocation and inversion events that have resulted in the duplication and truncation of some genes and have also given rise to new genes . There are 32 predicted pseudogenes, most of which are truncated fragments of genes associated with repeats . Rather then being the result of the reductive evolution seen in other intracellular bacteria, these pseudogenes appear to be the product of ongoing sequence duplication events. Chemistry . 2005 Jan 5; {Epub ahead of print} Structural Characterization of Artificial Self-Assembling Porphyrins That Mimic the Natural Chlorosomal Bacteriochlorophylls c, d, and e; Balaban TS et al.; We report two crystal structures of a synthetic porphyrin molecule which was programmed for self-assembly . The same groups which ensure that bacteriochlorophylls c, d, and e can self-assemble into the chlorosomal nanorods, the photosynthetic antenna system of some green bacteria, have been engineered into desired positions of the tetrapyrrolic macrocycle . In the case of the 5,15-meso-substituted anchoring groups, depending upon the concentration, by using the same crystallization solvents, either a tetragonal or a layered structure of porphyrin stacks were encountered . Surprinsingly, pi-pi interactions combined with extensive dispersive interactions, which also encompass cyclohexane, one of the crystallization solvents, win over putative hydrogen bonding . We are aware that our compounds differ considerably from the natural bacteriochlorophylls, but based upon our findings, we now question the hydrogen-bonding network, previously proposed to organize stacks of bacteriochlorophylls . Transmission electron microscopy (TEM), atomic force microscopy (AFM), and small-angle X-ray scattering (SAXS) on various isomeric compounds support our challenge of current models for the chlorosomal antenna as these show structures, astonishingly similar to those of chlorosomes. J Clin Microbiol, 2005 Jan, 43(1), 455 - 7 High prevalence of respiratory viral infections in patients hospitalized in an intensive care unit for acute respiratory infections as detected by nucleic acid-based assays; Legoff J et al.; Forty-seven bronchoalveolar lavages (BAL) were obtained from 41 patients with acute pneumonia attending an intensive care unit . By molecular diagnosis, 30% of total BAL and 63% of bacteria-negative BAL were positive for respiratory viruses . Molecular detection allows for high-rate detection of respiratory viral infections in adult patients suffering from severe pneumonia. J Clin Microbiol, 2005 Jan, 43(1), 229 - 34 Comparison of Helicobacter spp . in Cheetahs (Acinonyx jubatus) with and without gastritis; Terio KA et al.; Chronic gastritis causes significant morbidity and mortality in captive cheetahs but is rare in wild cheetahs despite colonization by abundant spiral bacteria . This research aimed to identify the Helicobacter species that were associated with gastritis in captive cheetahs but are apparently commensal in wild cheetahs . Helicobacter species were characterized by PCR amplification and sequencing of the 16S rRNA, urease, and cagA genes and by transmission electron microscopy of frozen or formalin-fixed paraffin-embedded gastric samples from 33 cheetahs infected with Helicobacter organisms (10 wild without gastritis and 23 captive with gastritis) . Samples were screened for mixed infections by denaturant gel gradient electrophoresis of the 16S rRNA gene and by transmission electron microscopy . There was no association between Helicobacter infection and the presence or severity of gastritis . Eight cheetahs had 16S rRNA sequences that were most similar (98 to 99%) to H . pylori . Twenty-five cheetahs had sequences that were most similar (97 to 99%) to "H . heilmannii" or H . felis . No cheetahs had mixed infections . The ultrastructural morphology of all bacteria was most consistent with "H . heilmannii," even when 16S rRNA sequences were H . pylori-like . The urease gene from H . pylori-like bacteria could not be amplified with primers for either "H . heilmannii" or H . pylori urease, suggesting that this bacteria is neither H . pylori nor "H . heilmannii." The cagA gene was not identified in any case . These findings question a direct role for Helicobacter infection in the pathogenesis of gastritis and support the premise that host factors account for the differences in disease between captive and wild cheetah populations. J Exp Biol, 2005 Jan 15, 208(Pt 2), 383 - 390 Redox signaling in colonial hydroids: many pathways for peroxide; Blackstone NW et al.; Studies of mitochondrial redox signaling predict that the colonial hydroids Eirene viridula and Podocoryna carnea should respond to manipulations of reactive oxygen species (ROS) . Both species encrust surfaces with feeding polyps connected by networks of stolons; P . carnea is more `sheet-like' with closely spaced polyps and short stolons, while E . viridula is more `runner-like' with widely spaced polyps and long stolons . Treatment with the chemical antioxidant vitamin C diminishes ROS in mitochondrion-rich epitheliomuscular cells (EMCs) and produces phenotypic effects (sheet-like growth) similar to uncouplers of oxidative phosphorylation . In peripheral stolon tips, treatment with vitamin C triggers a dramatic increase of ROS that is followed by tissue death and stolon regression . The enzymatic anti-oxidant catalase is probably not taken up by the colony but, rather, converts hydrogen peroxide in the medium to water and oxygen . Exogenous catalase does not affect ROS in mitochondrion-rich EMCs, but does increase the amounts of ROS emitted from peripheral stolons, resulting in rapid, runner-like growth . Treatment with exogenous hydrogen peroxide increases ROS levels in stolon tips and results in somewhat faster colony growth . Finally, untreated colonies of E . viridula exhibit higher levels of ROS in stolon tips than untreated colonies of P . carnea . ROS may participate in a number of putative signaling pathways: (1) high levels of ROS may trigger cell and tissue death in peripheral stolon tips; (2) more moderate levels of ROS in stolon tips may trigger outward growth, inhibit branching and, possibly, mediate the redox signaling of mitochondrion-rich EMCs; and (3) ROS may have an extra-colony function, perhaps in suppressing the growth of bacteria. Int Dent J, 2004 Dec, 54(6), 429 - 37 A review of the clinical efficacy of the Oral-B oscillating/rotating power toothbrush and the Philips Sonicare toothbrush in normal subject populations; Warren PR et al.; Plaque removal by a toothbrush results from a physical scrubbing of bristles on the tooth surface that removes adherent plaque bacteria . Because of the frequency of brush head motion, some power toothbrushes generally remove plaque more effectively than a manual brush . One power toothbrush, Philips Sonicare, claims also to remove plaque as a result of dynamic fluid activity . This effect has been shown in laboratory studies but clinical evidence is currently lacking . This review evaluated the data from well-controlled clinical studies carried out in normal subjects from a general population comparing the Sonicare toothbrushes with the Oral-B oscillating/rotating power toothbrush technology . It focuses on plaque removal from approximal surfaces where it is difficult for toothbrush bristles to reach, as it is here that any dynamic fluid effect should be most apparent . Results from the review found no evidence to support a greater efficacy for the Sonicare toothbrushes either generally or at approximal surfaces . Data revealed that the oscillating/rotating toothbrush was more effective than the Sonicare toothbrushes with respect to plaque removal . It is possible that factors associated with the clinical situation such as damping resulting from bristle contact with the tooth surface and the high viscosity of saliva and dentifrice may counteract dynamic fluid activity in vivo . This review indicates that dynamic fluid activity beyond the reach of bristles as demonstrated in the laboratory is yet unproven in the clinical situation. Microbiology, 2005 Jan, 151(Pt 1), 121 - 33 Molecular analysis of the anaerobic rumen fungus Orpinomyces - insights into an AT-rich genome; Nicholson MJ et al.; The anaerobic gut fungi occupy a unique niche in the intestinal tract of large herbivorous animals and are thought to act as primary colonizers of plant material during digestion . They are the only known obligately anaerobic fungi but molecular analysis of this group has been hampered by difficulties in their culture and manipulation, and by their extremely high A+T nucleotide content . This study begins to answer some of the fundamental questions about the structure and organization of the anaerobic gut fungal genome . Directed plasmid libraries using genomic DNA digested with highly or moderately rich AT-specific restriction enzymes (VspI and EcoRI) were prepared from a polycentric Orpinomyces isolate . Clones were sequenced from these libraries and the breadth of genomic inserts, both genic and intergenic, was characterized . Genes encoding numerous functions not previously characterized for these fungi were identified, including cytoskeletal, secretory pathway and transporter genes . A peptidase gene with no introns and having sequence similarity to a gene encoding a bacterial peptidase was also identified, extending the range of metabolic enzymes resulting from apparent trans-kingdom transfer from bacteria to fungi, as previously characterized largely for genes encoding plant-degrading enzymes . This paper presents the first thorough analysis of the genic, intergenic and rDNA regions of a variety of genomic segments from an anaerobic gut fungus and provides observations on rules governing intron boundaries, the codon biases observed with different types of genes, and the sequence of only the second anaerobic gut fungal promoter reported . Large numbers of retrotransposon sequences of different types were found and the authors speculate on the possible consequences of any such transposon activity in the genome . The coding sequences identified included several orphan gene sequences, including one with regions strongly suggestive of structural proteins such as collagens and lampirin . This gene was present as a single copy in Orpinomyces, was expressed during vegetative growth and was also detected in genomes from another gut fungal genus, Neocallimastix. J Biol Chem . 2005 Jan 4; {Epub ahead of print} Flux of fatty acids through NPC1 lysosomes; Passeggio J et al.; Niemann-Pick type C (NPC) is an autosomal recessive lipid storage disorder characterized by lysosomal accumulation of cholesterol and gangliosides resulting from a defect in intracellular lipid trafficking . The NPC1 gene encodes a 1278 amino acid integral membrane protein involved in the sub-cellular trafficking of lipids . The exact biological function of NPC1 remains unclear . Recent evidence suggests that NPC1 is a eukaryotic member of the RND permease family of transport proteins, which when expressed in bacteria is capable of transporting fatty acids . The goal of this project is to assess the role of NPC1 in the transport of fatty acids in primary human fibroblasts using normal fibroblasts and fibroblasts from patients with three lysosomal storage diseases: NPC, mucolipidosis IV, and Sandhoff's disease . If NPC1 is a fatty acid transporter we expect to find fatty acid accumulation only in NPC fibroblasts . We used three experimental approaches to assess the role of NC1 as a fatty acid transporter . First, we evaluated the accumulation versus metabolism of LDL-derived oleic acid . Second, the amount of free fatty acid present after growth in lipoprotein containing media was assessed . Third, we assessed the cellular accumulation of acriflavine, a fluorescent substrate for a number of RND permease transporters . Our results indicate that fatty acid flux through NPC1 deficient lysosomes is normal. J Biol Chem . 2005 Jan 4; {Epub ahead of print} Dimeric trigger factor stably binds folding-competent intermediates and cooperates with the DnaK-DnaJ-GrpE chaperone system to allow refolding; Liu CP et al.; Trigger factor (TF) is the first chaperone encountered by the nascent chain in bacteria and forms a stoichiometric complex with the ribosome . However, the functional significance of the high cytosolic concentration of uncomplexed TF, the majority of which is dimeric, is unknown . To gain insight into TF function, we investigated the TF concentration dependence of the GAPDH reactivation yield in the presence and absence of the DnaK-DnaJ-GrpE chaperone system in vitro . Cross-linking results indicate that the observed decrease in the reactivation yield of GAPDH at high concentrations of TF is due to the formation of a stable complex between TF dimer and GAPDH intermediates . In the absence of TF, or at low TF concentrations, the DnaK-DnaJ-GrpE chaperone system had negligible effect on the GAPDH refolding yield . However, GAPDH intermediates bound and held by dimeric TF could be specifically rescued by the DnaK-DnaJ-GrpE chaperone system in an ATP-dependent manner . This indicates the potential of TF, in its dimeric form, to act as a binding chaperone, maintaining non-native proteins in a refolding competent conformation and cooperating with down stream molecular chaperones to facilitate post-translational or post-stress protein folding. Nitric Oxide, 2005 Feb, 12(1), 46 - 53 Epub 2004 Dec 21. Nitric oxide synthase inhibitors and nitric oxide donors modulate the biosynthesis of thaxtomin A, a nitrated phytotoxin produced by Streptomyces spp; Wach MJ et al.; Evidence for the involvement of a bacterial nitric oxide synthase (NOS) in the biosynthesis of a phytotoxin is presented . Several species of Streptomyces bacteria produce secondary metabolites with unusual nitrogen groups, such as thaxtomin A (ThxA), which contains a nitroindole moiety . ThxA is a phytotoxin made by three pathogenic Streptomyces species that cause common scab of potato . All three species possess a gene homologous to the oxygenase domain of murine inducible NOS, and this gene, nos, is essential for normal levels of ThxA production . We grew Streptomyces turgidiscabies in the presence of several known NOS inhibitors and a nitric oxide (NO) scavenger to determine their effect on ThxA production . The NO scavenger (CPTIO) and four NOS inhibitors (NAME, NMMA, AG, and 7-NI) reduced ThxA production without affecting bacterial growth . A strain of S . turgidiscabies from which the nos gene had been deleted was grown in the presence of three NO donors (DEANO, SIN, and SNAP), and all three partially restored ThxA production . Our data suggest that bacterial nitric oxide synthases may, at least in part, produce NO for biosynthetic purposes, rather than for cellular signaling, as they do in mammals. Zhonghua Yi Xue Za Zhi, 2004 Nov 17, 84(22), 1879 - 82 {Comparison of efficacy between ceftriaxone and cefoperazone plus sulbactam in peri-operative treatment of acute suppurative cholangitis.}; Zhou XS et al.; OBJECTIVE: To compare the efficacy of ceftriaxone and that of cefoperazone plus sulbactam (sulperazon) in controlling infection, in scavenging bacteria from bile, and in their costs when treating acute suppurative cholangitis with choledochostomy . METHODS: Patients were randomly assigned to two groups: the ceftriaxone group (R-group, n = 95) and sulperazon group (S-group, n = 95) . Before choledochostomy, both groups received one intravenous dose of the corresponding antibiotics: and 2 g ceftriaxnoe for the R-group, 2 g sulperazon, containing 1 g cefoperazone and 1 g sulbactam, for the S-group . After the operation, the patients in the R-group received ceftriaxone 2 g i.v . q.d.; the patients in the S-group received sulperazon 2 g i.v . b.i.d. . In addition, all patients in both groups received metronidazole 0.5 g daily before and after the operation . The efficacy was evaluated by efficiency in controlling infection and the persisting days of symptoms due to infection, fever and leukocytosis; the persisting days was compared using the life table method to calculate the "cumulative probability of persistence of symptoms (CPPS)" . The two groups were also compared in regards to their biliary bacterial clearance rates and the costs directly attributable to the antibiotics . RESULTS: The efficiency in controlling infection was 98.9% (94/95) in both groups . However, the CPPS of the R-group decreased more rapidly than that of the S-group, Log-Rankchi(2) = 6.7901, P = 0.0092 . Biliary bacterial clearance rate on post-operative day 3 was 72.0% (36/50) for the R-group, 41.3% (19/46) for the S-group, P = 0.0037 . Cost directly attributable to the antibiotics were (1788.29 +/- 518.46) yuan (RMB) for the R-group, and (3768.74 +/- 820.55) yuan for the S-group, F = 395.51, P = 0.0000 . CONCLUSION: Both ceftriaxone and sulperazon are effective in treating acute suppurative cholangitis when used before and after choledochostomy . Ceftriaxone is superior in expediting symptom relief and bacterial clearance from bile, and is more cost-effective. Rocz Akad Med Bialymst, 2004, 49, 61 - 5 Cardiac surgical treatment of the patients with renal insufficiency; Hirnle T et al.; Cardiac surgical treatment of the patients with renal insufficiency became more frequent necessity . Also postoperative renal insufficiency occurs pretty often after cardiac surgery . That is in part a result of broadening of operative indications, which might concern patients with multiple diseases . Patients with renal insufficiency and coexistent heart diseases, patients with endocarditis and patients with renal insufficiency after cardiac surgery require the treatment of cardiac surgeons and nephrologists . Heart diseases are the main cause of the mortality in the dialysis patients . Among the patients with renal diseases the cardiac surgeon most often receive long-term dialysis patients with coexistent heart diseases, who needs cardiac surgery (coronary artery by-pass grafting, valve operations) . The amount of these operations increases, however it does not exceed 1% of overall number of cardiac operations . This group however, is very exacting and carries a high operative risk . Dialysis patients are exposed to increased risk of infection . 75% of them reveal infections in the form of sepsis . The presence of bacteria in the bloodstream increase the risk of infectious endocarditis . 6% of dialysis patients with IE require surgery . The prevention of renal failure after cardiac surgery is also very important . Renal insufficiency occurs in 12% of patients after cardiac surgery with the use of extracorporeal circulation . Renal failure complicates postoperative course and is of high risk for the patient . The mortality due to acute postoperative renal failure, which requires hemofiltration, reaches 70% . The proper cardiac surgical and nephrological management of renal insufficiency in patients selected for cardiac surgery as well as in patients with postoperative renal insufficiency is necessary to obtain good operative results. Appl Microbiol Biotechnol, 2005 Feb, 66(5), 475 - 85 Epub 2004 Nov 17. Cyclodextrin glucanotransferase: from gene to applications; Qi Q et al.; Cyclodextrin glucanotransferase (CGTase) is an important industrial enzyme which is used to produce cyclodextrins . CGTase genes from more than 30 bacteria have been isolated and several of the enzymes have been identified and biochemically characterized . For a better understanding of the reaction mechanism and function of CGTase, the enzyme has been analyzed at gene level and protein level with regard to its structure and the similarity of different CGTase subgroups . The biological role of the enzyme is proposed based on the genetic and enzymatic analyses . Methods to enhance the production of active CGTase by bacteria are compared . The enzyme can be applied in biotechnology for the production of cyclodextrins and oligosaccharides with novel properties. J Clin Invest, 2005 Jan, 115(1), 66 - 75 Human intestinal macrophages display profound inflammatory anergy despite avid phagocytic and bacteriocidal activity; Smythies LE et al.; Intestinal macrophages, which are thought to orchestrate mucosal inflammatory responses, have received little investigative attention compared with macrophages from other tissues . Here we show that human intestinal macrophages do not express innate response receptors, including the receptors for LPS (CD14), Fcalpha (CD89), Fcgamma (CD64, CD32, CD16), CR3 (CD11b/CD18), and CR4 (CD11c/CD18); the growth factor receptors IL-2 (CD25) and IL-3 (CD123); and the integrin LFA-1 (CD11a/CD18) . Moreover, resident intestinal macrophages also do not produce proinflammatory cytokines, including IL-1, IL-6, IL-10, IL-12, RANTES, TGF-beta, and TNF-alpha, in response to an array of inflammatory stimuli but retain avid phagocytic and bacteriocidal activity . Thus, intestinal macrophages are markedly distinct in phenotype and function from blood monocytes, although intestinal macrophages are derived from blood monocytes . To explain this paradox, we show that intestinal stromal cell-derived products downregulate both monocyte receptor expression and, via TGF-beta, cytokine production but not phagocytic or bacteriocidal activity, eliciting the phenotype and functional profile of intestinal macrophages . These findings indicate a mechanism in which blood monocytes recruited to the intestinal mucosa retain avid scavenger and host defense functions but acquire profound "inflammatory anergy," thereby promoting the absence of inflammation characteristic of normal intestinal mucosa despite the close proximity of immunostimulatory bacteria. Proc Natl Acad Sci U S A, 2005 Jan 11, 102(2), 373 - 8 Epub 2005 Jan 03. Phylogeny determined by protein domain content; Yang S et al.; A simple classification scheme that uses only the presence or absence of a protein domain architecture has been used to determine the phylogeny of 174 complete genomes . The method correctly divides the 174 taxa into Archaea, Bacteria, and Eukarya and satisfactorily sorts most of the major groups within these superkingdoms . The most challenging problem involved 119 Bacteria, many of which have reduced genomes . When a weighting factor was used that takes account of difference in genome size (number of considered folds), small-genome taxa were mostly grouped with their full-sized counterparts . Although not every organism appears exactly at its classical phylogenetic position in these trees, the agreement appears comparable with the efforts of others by using sophisticated sequence analysis and/or combinations of gene content and gene order . During the course of the study, it emerged that there is a core set of approximately 50 folds that is found in all 174 genomes and a single fold diagnostic of all Archaea. Peptides, 2005 Feb, 26(2), 291 - 6 Agrocybin, an antifungal peptide from the edible mushroom Agrocybe cylindracea; Ngai PH et al.; An antifungal peptide with a molecular mass of 9kDa was isolated from fresh fruiting bodies of the mushroom Agrocybe cylindracea . The isolation procedure comprised ion exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue gel, ion exchange chromatography by fast protein liquid chromatography (FPLC) on Mono S, and FPLC-gel filtration on a Superdex 75 column . The antifungal peptide, designated as agrocybin, was unadsorbed on DEAE-cellulose, and adsorbed on Affi-gel blue gel and Mono S . Agrocybin exerted antifungal activity against several fungal species but lacked inhibitory activity against bacteria when tested up to 300muM . The activity of HIV-1 reverse transcriptase was attenuated in the presence of agrocybin . It exhibited weaker mitogenic activity than Con A on isolated murine splenocytes, but was devoid of antiproliferative activity on Hep G2 (hepatoma) cells when tested at 110muM. Vaccine, 2005 Jan 19, 23(9), 1178 - 88 Identification of new potential vaccine candidates against Chlamydia pneumoniae by multiple screenings; Finco O et al.; Chlamydia are intracellular bacteria associated to serious human disease . A vaccine has proved difficult to obtain so far, and current opinions agree that multi-antigen combinations may be required to induce optimal protective responses . In order to identify new potential vaccine candidates, we recently screened the Chlamydia pneumoniae (Cpn) genome and described 53 recombinant proteins which elicited antibodies binding to purified Cpn cells . We now report that six proteins in this group can also induce in vitro neutralizing antibodies . Antibody specificity for the corresponding antigens was assessed by immunoblot analysis of 2DE Cpn protein maps . Furthermore, four of the six in vitro neutralizing antigens (Pmp2, Pmp10, OmpH-like and enolase) could inhibit Cpn dissemination in a hamster model . The results show that these Cpn proteins are immunoaccessible in infectious EBs, and recommend further investigation on their value as vaccine components. Med Arh, 2004, 58(5), 285 - 7 {Neutropenia febriles at children who have leukemia}; Sakic M; INTRODUCTION: Neutropenia febriles are the most frequent complications during the hospital treatment of children who are ill of malignant . It is urgent state end each of feverish should be treated as infection, until it is proved as opposite . It is necessary immediately to begin with antibiotical therapy according to the algoritham for therapy of neutropenia febriles . Neutropenia febriles is caused by virus, bacteria and fungus infection . RESULTS: It were 83 leukemia treated children on Hematooncology department of pediatrical Clinic in Sarajevo in period from 01.01.1997 until 31.12.2002 . Neutropenia febriles were seen at 41 of children (49.4%) . Between all of neutropenia febriles the causes were isolated at 27 children (65%) . The most of them had sepsis, and at 40% of children had positive hemo-cultures . In the hemo-cultures the gr . positive causes were isolated at the 55% of children, gr . negative causes were isolated at 25% of children, and fungus were isolated at 20% . Between the other cultures urino-cultures were positive at 18.5% of children . RTG pulmo was isolated at 18.5%, smear pharyngs was isolated at 11% . CONCLUSION: In such state of patient it should be thought about the possibility of contamination of patient and protect him/her from superinfection, by him/her in isolation box and staff should take care about sepsis and antisepsis. Pol Merkuriusz Lek, 2004 Sep, 17(99), 284 - 8 {Can we be infected with atherosclerosis?}; Rusiecka E; Atherosclerosis is one of the most important problems of the contemporary medicine and it is the most frequent reason of deaths in Western countries . Conventional risk factors are responsible for only 50% cases of atherosclerosis . Investigations performed over last 20 years revealed new risk factors, like for example bacterial and viral infections . In recent scientific descriptions Chlamydia pneumoniae, Helicobacter pylori and viruses from Herpesviridae family (Cytomegalovirus hominis, Epstein-Barr virus, Herpesvirus hominis) are mentioned as those, which could take part in the atherogenesis . The infectious theory of atherosclerosis might be proved by the presence of maintained above pathogens in the atheromatous plaque and the existence of antibodies against bacteria and viruses in blood. J Immunol Methods, 2004 Dec, 295(1-2), 119 - 27 Epub 2004 Oct 27. Direct fluorochrome labeling of phage display library clones for studying binding specificities: applications in flow cytometry and fluorescence microscopy; Jaye DL et al.; Phage display technology is increasingly employed to identify high-affinity peptides and single-chain antibodies with binding specificities for a diversity of target types . The analysis of phage-binding sensitivity and specificity typically employs directly labeled secondary antiphage antibodies and potentially tertiary labels, such as fluorochromes and enzymes, when biotinylated antibodies are used . However, secondary or tertiary reagents may not be feasible or desirable for some target types and applications . Here, we present a simple approach for directly labeling phage clones with two common amine-reactive fluorochromes . We show that these fluorochromes label the pVIII major coat protein and that the binding selectivity of peptides displayed on the pIII protein of several well-characterized phage clones is maintained in flow cytometric analysis and immunofluorescence microscopy . Uniquely, such labeled phage, in part, represent self-propagating reagents because conjugation does not impair the ability to efficiently reproduce in bacteria, although relabeling with fluorochrome would be necessary . Our data suggest that primary labeled phage clones may be used similarly to primary antibody conjugates. Biochemistry (Mosc), 2004 Dec, 69(12), 1399 - 402 Effect of Low Temperatures on Photochemical Activity of PS1 Reaction Centers from Synechocystis sp . Frozen under Illumination; Knox PP et al.; After cooling of Synechocystis sp . photosystem 1 (PS1) reaction centers (RC) to 160 K under illumination most of the photoactive pigment is fixed for a long time in the oxidized state . The same effect is observed in purple bacteria RC . The dark reduction kinetics of PS1 P700 chlorophyll, which still retains its photochemical activity, in these samples was similar to that in samples cooled in the dark . We suggest that the photoinduced charge separation in PS1 RC, as well as in purple bacteria RC, is accompanied by conformational changes that can be fixed in samples cooled under illumination . As a result, the electrons photomobilized in RC cooled under illumination are unable to return backward the process of electron transfer to P700(+) after cessation of actinic illumination . Such irreversible trapping of electrons can take place in different parts of the PS1 RC electron acceptor chain. J Biochem (Tokyo), 2004 Oct, 136(4), 415 - 9 Bordetella Dermonecrotic Toxin Exerting Toxicity through Activation of the Small GTPase Rho; Fukui A et al.; Bordetella dermonecrotic toxin (DNT) is a virulence factor produced by bacteria belonging to the genus Bordetella . The toxin possesses novel transglutaminase activity that catalyzes polyamination or deamidation of the small GTPases of the Rho family . The modified GTPases loose their GTP hydrolyzing activity, function as a constitutive active molecule, and continuously transduce signals to downstream effectors, which mediate the consequent phenotypes of cells intoxicated by DNT . A dynamin-dependent endocytosis is required for the toxin to be internalized into cells although it is unlikely transported to deep organelles such as the Golgi apparatus or the ER . Several lines of evidence show that the toxin undergoes proteolytic cleavage by furin or furin-like protease probably in the early endosome, and then escapes into the cytoplasm to reach the GTPase. Acta Paediatr Taiwan, 2004 Jul-Aug, 45(4), 249 - 52 Recurrent multiple hepatic abscesses, hepatic calcification and congenital hearing loss in a ch