Water Sci Technol, 2002, 45(1), 25 - 32 Compartmental model for organic matter digestion in facultative ponds; Giraldo E et al.; A model has been developed for the digestion of organic matter in facultative ponds in tropical regions . Complete mixing has been assumed for the aerobic and anaerobic compartments . Settling, aerobic layer oxidation, and anaerobic layer methanogenesis are the main processes for organic matter removal in the water column . Exchange processes between layers are dispersive or soluble exchange, solubilization and transport of organic matter from sediments to water column are also taken into account . Degradation of organic matter in the sediments produces gaseous emissions to the water column . The exchange between bubbles ascending and the water column was measured . The model was calibrated with data obtained from a pilot facultative pond built in Muna Reservoir in Bogota . The pond was sampled during 4 months to compare data between its water hyacinth covered section and uncovered section . The results clearly show the relative importance of different BOD removal processes in facultative ponds and suggest modifications to further improve performance . The results from the model suggest that internal loadings to facultative ponds due to solubilization and return of organic matter from the sediments to the aerobic layer greatly influence the soluble BOD effluent concentration . Aerobic degradation activity in the facultative pond does not affect significantly the effluent concentration . Anaerobic degradation activity in the facultative pond can more easily achieve increases in the removal efficiencies of BOD. Methods Mol Biol, 2002, 172, 21 - 49 Calcium; Williams RJ; This chapter describes the chemical and biological value of the calcium ion . In calcium chemistry, our main interest is in equilibria within static, nonflowing systems . Hence, we examined the way calcium formed precipitates and complex ions in solution . We observed thereafter its uses by humankind in a vast number of materials such as minerals, e.g., marble, concrete, mortars, which parallel the biological use in shells and bones . In complex formation, we noted that many combinations were of anion interaction with calcium for example in the uses of detergents and medicines . The rates of exchange of calcium from bound states were noted but they had little application . Calcium ions do not act as catalysts of organic reactions . In biological systems, interest is in the above chemistry, but extends to the fact that Ca2+ ions can carry information by flowing in one solution or from one solution to another through membranes . Hence, we became interested in the details of rates of calcium exchange . The fast exchange of this divalent ion from most organic binding sites has allowed it to develop as the dominant second messenger . Now the flow can be examined in vitro as calcium binds particular isolated proteins, which it activates as seen in physical mechanical changes or chemical changes and this piece-by-piece study of cells is common . Here, however, we have chosen to stress the whole circuit of Ca2+ action indicating that the cell is organized both at a basal and an activated state kinetic level by the steady state flow of the ion (see Fig . 11) . Different time constants of exchange utilizing very similar binding constants lead to: 1) fast responses as in the muscle of an animal; or 2) slower change as in differentiation of an egg or seed . Many other changes of state may relate to Ca2+ steady-state levels of flow in the circuitry and here we point to two: 1) dormancy in reptiles and animals; and 2) sporulation in both bacteria and lower plants . In the other chapters of this volume many components of the overall circuitry will be described . The reader should try to marry these into the overall activity of the cell for on top of molecular biology there is the cooperative system molecular biology of cells . To give an analogy, whereas much can be understood from the analysis of the properties of single-isolated water molecules, even examining their interaction in ice, this study alone cannot lead to an appreciation of the melting or boiling points of bulk water. Int J Hyg Environ Health, 2001 Dec, 204(4), 245 - 9 A case report of false negative Legionella test results in a chlorinated public hot water distribution system due to the lack of sodium thiosulfate in sampling bottles; Wiedenmann A et al.; We examined samples from the showers and the central water distribution system of a public building with an indoor swimming pool . The pool was used for school and recreational activities and as a sports therapy facility for patients with coronary heart disease . The building's hot water system was contaminated with Legionella pneumophila . Due to the building's intricate piping system, several attempts to completely eliminate legionellae by thermal and chemical disinfection had failed, so an external sanitation company was charged with the installation of a continuous chlorination device in order to keep Legionella concentrations low . The laboratory which was contracted by the sanitation company to monitor bacteria levels after installation of the chlorination device used sampling bottles without sodium thiosulfate and repeatedly reported an absence of Legionella . However, up to 69,000 colony forming particles (CFP) of Legionella pneumophila (Lp) per litre and up to 171 CFP/ml of heterotrophic bacteria could be detected when parallel samples were collected in bottles containing sodium thiosulfate at standard concentrations . Laboratories, epidemiologists, public health officials and technical staff who may be in charge of delivering, preparing or using sterile sampling devices for the collection of environmental samples to be tested for legionellae should be aware that cultures can return false negative results if the sampling containers used to collect chlorinated drinking water or chlorinated pool water samples do not contain a neutralizing agent to instantly inactivate residual halogen biocides . False negative results may lead to a false sense of security regarding the safety of water systems or the success of disinfection measures, and may thus endanger public health or even hinder the epidemiological clarification of outbreaks. Zhonghua Wai Ke Za Zhi, 2000 Dec, 38(12), 895 - 6 {CT guided percutaneous fine needle aspiration for early diagnosis of pancreatic infection after acute necrotizing pancreatitis}; Li N et al.; OBJECTIVE: To assess CT-guided percutaneous fine needle aspiration (FNA) for early diagnosis of pancreatic infection after acute necrotizing pancreatitis . METHODS: CT-guided percutaneous FNA was employed to diagnose pancreatic infection after acute necrotizing pancreatitis in 14 patients . The aspirates were smeared, cultured and subjected polymerase to chain reaction (PCR) for detecting bacteria . RESULTS: Ten patients were diagnosed as having pancreatic infection and 4 non-pancreatic infection . FNA was totally performed for 15 times: left retroperitoneal route for 12 times, left and right retroperitoneal route for one time, and transperitoneal route for two times . The sensitivity of smear, PCR, culture to diagnose pancreatic infection was 7/10, 9/10, 10/10, respectively . Their specificity was 4/4, and the time for obtaining the results was 30 minutes, 4 hours, 3 days respectively . CONCLUSIONS: Retroperitoneal approach can be used for most patients with acute necrotizing pancreatitis when performing CT guided-percutaneous FNA . PCR of aspirates is a rapid and accurate method to diagnose pancreatic infection. Comp Immunol Microbiol Infect Dis, 2002 Jan, 25(1), 21 - 7 An immunochromatographic serological assay for the diagnosis of Mycobacterium tuberculosis; Reddy JR et al.; A rapid serological test for tuberculosis (TB) infection was designed using antigens specific to Mycobacterium tuberculosis . Tuberculosis infection, TB vaccination and exposure to environmental Mycobacteria cannot be distinguished using skin tests based on tuberculin protein derivatives . The standard diagnostic techniques such as skin tests, X-rays and DNA techniques are time consuming, expensive, and not practical for screening large populations . We used the 38, 63, 64, 14, 59-kDa antigens of M . tuberculosis to develop a rapid immunochromatographic test kit . This study evaluates the diagnostic potential of the rapid test kit using TB positive and TB negative serum samples from various hospitals in India . The samples were obtained from patients infected with or exposed to bacteria and viral pathogens . The results demonstrated that the combination of antigens improved the diagnostic specificity and sensitivity . The specificity of the test was 99.42% with sensitivity of 98.52% (n = 241) . In case of multiple infections, the specificity was 93.15% with a low sensitivity of 73.52% n = 141) . The test kit may offer an improved alternative to purified protein derivative (PPD) . This rapid TB test kit may be a useful tool for first-line testing of suspected cases, epidemiological studies and in designing a quality health system to reduce health hazards in resource-poor countries. Lab Anim, 2002 Jan, 36(1), 43 - 50 Airborne contaminants in conventional laboratory rabbit rooms; Kaliste E et al.; Besides the well known allergens, several other risk factors may exist for health in a laboratory animal unit . The exposure to these factors may be significant in animal units with poor general or local ventilation systems . Moreover, means to prevent the distribution of airborne contaminants may be limited in animal units housing rabbits or other bigger laboratory animals . Airborne contaminants in conventional laboratory rabbit rooms were sought to evaluate the occupational exposure of animal care personnel . Concentrations of airborne dust, bacteria, fungi, ammonia and endotoxins were measured during 2 days in three phases: before working activities began, during them and afterwards . Both stationary and some personal samples were taken . All of the contaminants sought were found in the rabbit room air . When compared to reported levels in farm animal production areas, the concentrations measured were generally low . However, moderate or high levels of airborne bacteria and fungi were found occasionally during work routines . Airborne contaminants should be considered as a potential occupational health risk for persons working with laboratory animals. Appl Microbiol Biotechnol, 2002 Jan, 58(1), 13 - 22 Quinoproteins: structure, function, and biotechnological applications; Matsushita K et al.; A new class of oxidoreductase containing an amino acid-derived o-quinone cofactor, of which the most typical is pyrroloquinoline quinone (PQQ), is called quinoproteins, and has been recognized as the third redox enzyme following pyridine nucleotide- and flavin-dependent dehydrogenases . Some quinoproteins include a heme c moiety in addition to the quinone cofactor in the molecule and are called quinohemoproteins . PQQ-containing quinoproteins and quinohemoproteins have a common structural basis, in which PQQ is deeply embedded in the center of the unique superbarrel structure . Increased evidence for the structure and function of quinoproteins has revealed their unique position within the redox enzymes with respect to catalytic and electron transfer properties, and also to physiological and energetic function . The peculiarities of the quinoproteins, together with their unique substrate specificity, have encouraged their biotechnological application in the fields of biosensing and bioconversion of useful compounds, and also to environmental treatment. Ned Tijdschr Tandheelkd, 1994 Jun, 101(6), 231 - 7 {Amalgam . XV . Electrochemical activity in the mouth}; Boere G; Almost all metals used in dentistry can corrod . The uniform corrosion is not so important in concern to the used metals . This type of corrosion may become important in an extreme environment with the low pH which can exist in plaque, denture adhesive and fluoride gel . An important type of corrosion is contact corrosion, which results in a higher corrosion rate of the base metal in the galvanic couple noble-base metal . As a result of the small io (exchange current dentistry) the higher corrosion rate is the result of a larger reaction surface for the reduction reaction . Another important type of corrosion is caused by differences in the surface . As a result of this differentiation pitting or crevice corrosion may occur . Also bacteria and other micro-organisms can have an influence on the corrosion process . The corrosion rate is not only determined by the redoxpotential but also by other factors. J Microbiol Methods, 2002 Apr, 49(2), 103 - 19 Exposing culprit organic pollutants: a review; Keane A et al.; There is a continuing need for monitoring the health of the environment due to the presence of pollutants . Here, we review the development and attributes of biosensors by which bacteria have been genetically modified to express the luminescence genes, i.e . to glow, in a quantified manner, in response to pollutants . We have concentrated on the detection of organic hydrocarbon pollutants and discussed the molecular mechanisms by which some of these chemicals act as effector molecules on the respective regulatory systems . The future of environmental biosensors is predictably bright . As more knowledge is gathered on the sensing regulatory component, the possibility of developing targeted or pollutant-specific biosensors is promising . Moreover, the repertoire of biosensors for culprit organic pollutants is expected to be enlarged through advances in genomics technology and identification of new sensory or receptor molecules . The need for pollutant detection at concentrations in the parts per trillion range or biosensors configured in a nanoscale is anticipated. Phytochemistry, 2002 Feb, 59(3), 279 - 91 Serine proteinase inhibitors in the Compositae: distribution, polymorphism and properties; Konarev AV et al.; Multiple molecular forms of inhibitors of trypsin (TI) and chymotrypsin (CI), which are typical digestive enzymes of insects, mammals and micro-organisms, and subtilisin (SI), a proteinase of many bacteria and phytopathogenic fungi, were identified in seeds and vegetative organs of the majority of 128 wild and cultivated species representing 65 genera of three of the subfamilies of the Compositae . Inhibitors with M(r) ranging from 7450 to 7800 and combining activities towards subtilisin and trypsin and/or chymotrypsin (T/C/SI) had the widest distribution and may be involved in plant defense mechanisms . They were found in many species of the subfamilies Carduoideae (genera Carthamus, Centaurea, Cirsium), Cichorioideae (Lactuca, Taraxacum) and Asteroideae (Helianthus, Cosmos, Bidens) . Partial amino acid sequencing showed that the safflower (Carthamus tinctorius) T/C/SI and Cosmos bipinnatus T/C/SI, T/SI and C/SI belonged to the potato I inhibitor family . The most active, variable and heterogeneous inhibitors were found in species of the tribe Heliantheae, which is placed in the evolutionary advanced subfamily Asteroideae . Seeds of Helianthus species, Eclipta prostrata, Gailardia aristata, Zinnia elegans and Silphium perfoliatum contained various TI with M(r) ranging from 1500 to 14,750, with some also containing SI . H . annuus seeds contain a unique cyclic TI of M(r) 1514 and similar TI were also present in other Helianthus spp . and the related species Tithonia diversifolia . Zinnia elegans contained a TI with M(r) 11,350 which appeared to represent a novel type of inhibitor distantly related to the cereal subgroup of Bowman-Birk inhibitors . TI and T/SI varied widely in H . annuus lines and wild Helianthus species in their presence or absence and composition . Similar T/SI components were found in the cultivated diploid H . annuus and annual diploid species with the B genome but not in perennials with the A genome . Some T/SI, SI and TI were detected in vegetative organs of sunflower and other Compositae . Studies of the polymorphism and distribution of proteinase inhibitors are relevant to the evolution of protective protein systems and the mechanisms of resistance to pathogenic organisms in the Compositae and other plants. Biochem Biophys Res Commun, 2002 Feb 15, 291(1), 158 - 64 Interaction network of human aminoacyl-tRNA synthetases and subunits of elongation factor 1 complex; Sang Lee J et al.; Aminoacyl-tRNA synthetases (ARSs) ligate amino acids to their cognate tRNAs . It has been suggested that mammalian ARSs are linked to the EF-1 complex for efficient channeling of aminoacyl tRNAs to ribosome . Here we systemically investigated possible interactions between human ARSs and the subunits of EF-1 (alpha, beta, gamma, and delta) using a yeast two-hybrid assay . Among the 80 tested pairs, leucyl- and histidyl-tRNA synthetases were found to make strong and specific interaction with the EF-1gamma and beta while glu-proly-, glutaminyl-, alanyl-, aspartyl-, lysyl-, phenylalanyl-, glycyl-, and tryptophanyl-tRNA synthetases showed moderate interactions with the different EF-1 subunits . The interactions of leucyl- and histidyl-tRNA synthetase with the EF-1 complex were confirmed by immunoprecipitation and in vitro pull-down experiments . Interestingly, the aminoacylation activities of these two enzymes, but not other ARSs, were stimulated by the cofactor of EF-1, GTP . These data suggest that a systematic interaction network may exist between mammalian ARSs and EF-1 subunits probably to enhance the efficiency of in vivo protein synthesis . (c)2002 Elsevier Science (USA). Acupunct Med, 2001 Dec, 19(2), 112 - 6 Skin disinfection and acupuncture; Hoffman P; The need for skin disinfection before insertion of an acupuncture needle is controversial and there is no specific research on this topic . However research and observations on the effect of, and the need for, skin disinfection before injections forms a good analogy of acupuncture . Whilst micro-organisms present on the surface of the skin are accessible to disinfection, those located under the surface in ducts, glands and follicles are out of reach and can be inoculated into the sterile tissues below by needle insertion . Fortunately, the bacteria resident on the skin have a low potential to cause infection if host immunity is not severely impaired or compromised by the long-term presence of foreign material, such as a surgical stitch . Disinfection of clean skin before injection is not generally considered necessary and observations of lack of infection following injections without prior skin disinfection support this; however, contamination by micro-organisms not normally resident on skin can pose a higher risk of infection . If skin is visibly soiled, it should be washed and if needle insertion is near an infected or contaminated site, it should be disinfected with alcohol . Practitioner hand hygiene between patients is important, even if gloves are worn . Hands should be washed with soap or detergent and water, or an alcohol handrub can be used if hands are physically clean. J Cell Biol, 2002 Feb 4, 156(3), 511 - 8 Epub 2002 Feb 04. Visualization of Rab9-mediated vesicle transport from endosomes to the trans-Golgi in living cells; Barbero P et al.; Mannose 6-phosphate receptors (MPRs) are transported from endosomes to the trans-Golgi via a transport process that requires the Rab9 GTPase and the cargo adaptor TIP47 . We have generated green fluorescent protein variants of Rab9 and determined their localization in cultured cells . Rab9 is localized primarily in late endosomes and is readily distinguished from the trans-Golgi marker galactosyltransferase . Coexpression of fluorescent Rab9 and Rab7 revealed that these two late endosome Rabs occupy distinct domains within late endosome membranes . Cation-independent mannose 6-phosphate receptors are enriched in the Rab9 domain relative to the Rab7 domain . TIP47 is likely to be present in this domain because it colocalizes with the receptors in fixed cells, and a TIP47 mutant disrupted endosome morphology and sequestered MPRs intracellularly . Rab9 is present on endosomes that display bidirectional microtubule-dependent motility . Rab9-positive transport vesicles fuse with the trans-Golgi network as followed by video microscopy of live cells . These data provide the first indication that Rab9-mediated endosome to trans-Golgi transport can use a vesicle (rather than a tubular) intermediate . Our data suggest that Rab9 remains vesicle associated until docking with the Golgi complex and is rapidly removed concomitant with or just after membrane fusion. J Biol Chem, 2002 May 3, 277(18), 15962 - 70 Epub 2002 Feb 04. Interaction of fibroblast growth factor receptor 3 and the adapter protein SH2-B . A role in STAT5 activation; Kong M et al.; Fibroblast growth factor receptor 3 (FGFR3) influences a diverse array of biological processes, including cell growth, differentiation, and migration . Activating mutations in FGFR3 are associated with multiple myeloma, cervical carcinoma, and bladder cancer . To identify proteins that interact with FGFR3 and which may mediate FGFR3-dependent signaling, a yeast two-hybrid screen was employed using the cytoplasmic kinase domain of FGFR3 as bait . We identified the adapter protein SH2-B as an FGFR3-interacting protein . Coimmunoprecipitation experiments demonstrate binding of the SH2-B beta isoform to FGFR3 in 293T cells . Tyrosine phosphorylation of SH2-B beta was observed when coexpressed with activated FGFR3 mutants such as the weakly activated mutant N540K or the strongly activated mutant K650E, both associated with human developmental syndromes . The extent of tyrosine phosphorylation of SH2-B beta correlates with receptor activation, suggesting that FGFR3 activation mediates tyrosine phosphorylation of SH2-B beta . Furthermore, two tyrosine phosphorylation sites of FGFR3, Tyr-724 and Tyr-760, are required for optimal binding of the Src homology-2 (SH2) domain of SH2-B beta . We also demonstrate the phosphorylation and nuclear translocation of Stat5 by activated FGFR3, which increases in response to overexpression of SH2-B beta . Taken together, our results identify SH2-B beta as a novel FGFR3 binding partner that mediates signal transduction. Cardiovasc Res, 2002 Feb 1, 53(2), 405 - 13 Nitric oxide as a mediator of delayed pharmacological (A(1) receptor triggered) preconditioning; is eNOS masquerading as iNOS? Bell RM, Smith CC, Yellon DM. BACKGROUND: Nitric oxide (NO), synthesised from the inducible isoform of nitric oxide synthase (iNOS), is implicated in mediating second window of protection (SWOP)/delayed ischemic preconditioning . However the role of NO and iNOS in delayed pharmacological protection remains unclear and is the subject of this investigation . METHODS: To test the hypothesis that iNOS is necessary for delayed pharmacological preconditioning, the adenosine A(1) receptor agonist, 2-chloro N(6) cyclopentyl adenosine (CCPA) (25 microg/kg i.v.) or saline was administered to wild type (WT) or iNOS gene knockout mice (KO) . Twenty-four hours later, the hearts were isolated, Langendorff perfused and subjected to 35 min ischemia/30 min reperfusion prior to infarct size determination . RESULTS: WT and KO control hearts had identical infarct sizes of 37 +/- 3% and 37 +/- 2%, respectively . CCPA significantly reduced infarct size in WT hearts to 22 +/- 2% and also, unexpectedly, in KO hearts (27 +/- 2%) . This protection was abrogated with the non-specific NOS inhibitor, N(omega) nitro L-arginine methyl ester (L-NAME, 100 microM), and could be mimicked in naive hearts with the NO donor, donor S-nitroso N-acetyl DL penicillamine (SNAP, 1 microM) . Delayed protection appeared to be mediated by NO synthesis in both WT and KO hearts . Additional studies using Western blot analysis demonstrated endothelial NOS (eNOS) upregulation and increased NO(x) release in both WT and KO hearts . CONCLUSIONS: This is the first study to demonstrate a role for eNOS in delayed A(1) receptor triggered (pharmacological) preconditioning, potentially representing a new pharmacological target for protecting the ischemic heart. Biochemistry, 2002 Feb 12, 41(6), 1778 - 85 8-(Hydroxymethyl)-3,N(4)-etheno-C, a potential carcinogenic glycidaldehyde product, miscodes in vitro using mammalian DNA polymerases; Singer B et al.; 8-(Hydroxymethyl)-3,N(4)-etheno-C (8-HM-epsilonC) is an exocyclic adduct resulting from the reaction of dC with glycidaldehyde, a mutagen and animal carcinogen . This compound has now been synthesized and its phosphoramidite incorporated site-specifically into a defined 25-mer oligonucleotide . In this study, the mutagenic potential of this adduct in the 25-mer oligonucleotide was investigated in an in vitro primer-template extension assay using four mammalian DNA polymerases . The miscoding potentials were also compared to those of an analogous derivative, 3,N(4)-etheno C (epsilonC), in the same sequence . Both adducts primarily blocked replication by calf thymus DNA polymerase alpha at the modified base, while human polymerase beta catalyzed measurable replication synthesis through both adducts . Nucleotide insertion experiments showed that dA and dC were incorporated by pol beta opposite either adduct, which would result in a C --> T transition or C --> G transversion . Human polymerase eta, a product of the xeroderma pigmentosum variant (XP-V) gene, catalyzed the most efficient bypass of the two lesions with 25% and 32% for 8-HM-epsilonC and epsilonC bypassed after 15 min . Varying amounts of all four bases opposite the modified bases resulted with pol eta . Human polymerase kappa primarily blocked synthesis at the base prior to the adduct . However, some specific misincorporation of dT resulted, forming an epsilonC.T or 8-HM-epsilonC.T pair . From these data, we conclude that the newly synthesized glycidaldehyde-derived adduct, 8-HM-epsilonC, is a miscoding lesion . The bypass efficiency and insertion specificity of 8-HM-epsilonC and epsilonC were similar for all four polymerases tested, which could be attributed to the similar planarity and sugar conformations for these two derivatives as demonstrated by molecular modeling studies. Chemosphere, 2002 Jan, 46(2), 305 - 10 Prediction of mixture toxicity with its total hydrophobicity; Lin Z et al.; Based on the C18 Empore disk/water partition coefficient of a mixture, quantitative structure-activity relationships (QSARs) are presented, which are used to predict the toxicity of mixed halogenated benzenes to P . phosphoreum . The predicted toxicity of 10 other related mixtures based on the QSAR model, agree well with the observed data with r2 = 0.973, SE = 0.113 and F = 287.785 at a level of significance P < 0.0001 . The joint effect of these chemicals is simple similar action and the toxicity of the mixtures can be predicted from total hydrophobicity and is independent of hydrophobicity of the components or the ratio of the individual chemicals. Biosci Biotechnol Biochem, 2001 Dec, 65(12), 2763 - 72 Purification and characterization of membrane-bound quinoprotein cyclic alcohol dehydrogenase from Gluconobacter frateurii CHM 9; Moonmangmee D et al.; A quinoprotein catalyzing oxidation of cyclic alcohols was found in the membrane fraction for the first time, after extensive screening among aerobic bacteria . Gluconobacter frateurii CHM 9 was finally selected in this study . The enzyme tentatively named membrane-bound cyclic alcohol dehydrogenase (MCAD) was found to occur specifically in the membrane fraction, and pyrroloquinoline quinone (PQQ) was functional as the primary coenzyme in the enzyme activity . MCAD catalyzed only oxidation reaction of cyclic alcohols irreversibly to corresponding ketones . Unlike already known cytosolic NAD(P)H-dependent alcohol-aldehyde or alcohol-ketone oxidoreductases, MCAD was unable to catalyze the reverse reaction of cyclic ketones or aldehydes to cyclic alcohols . MCAD was solubilized and purified from the membrane fraction of the organism to homogeneity . Differential solubilization to eliminate the predominant quinoprotein alcohol dehydrogenase (ADH), and the subsequent two steps of column chromatographies, brought MCAD to homogeneity . Purified MCAD had a molecular mass of 83 kDa by SDS-PAGE . Substrate specificity showed that MCAD was an enzyme oxidizing a wide variety of cyclic alcohols . Some minor enzyme activity was found with aliphatic secondary alcohols and sugar alcohols, but not primary alcohols, differentiating MCAD from quinoprotein ADH . NAD-dependent cytosolic cyclic alcohol dehydrogenase (CCAD) in the same organism was crystallized and its catalytic and physicochemical properties were characterized . Judging from the catalytic properties of CCAD, it was apparent that CCAD was distinct from MCAD in many respects and seemed to make no contributions to cyclic alcohol oxidation. Pediatr Clin North Am, 2002 Feb, 49(1), 127 - 41 Probiotics in health and disease in the pediatric patient; Markowitz JE et al.; Probiotics are truly a timeless concept . Initially standard components of the human diet, potentially beneficial bacteria and yeast have been eliminated systematically through modern methods of preparing and preserving foods . Although the concept of probiotics is not new, the science of probiotics is in its infancy . Only recently have techniques been developed to identify, culture, and produce the probiotics that are suitable for medical use consistently . Furthermore, the potential uses for these organisms may extend far beyond what originally was thought . In no way should probiotics be thought of as a panacea for the diseases described earlier; however, when used appropriately, they represent a potentially beneficial adjunct to other proved therapies and have the added benefit of providing a stabilizing influence on the delicate balance between the ecosystem that consists of the human and its flora. Plant Cell, 2002 Jan, 14(1), 263 - 73 Repair of damaged DNA by Arabidopsis cell extract; Li A et al.; All living organisms have to protect the integrity of their genomes from a wide range of genotoxic stresses to which they are inevitably exposed . However, understanding of DNA repair in plants lags far behind such knowledge in bacteria, yeast, and mammals, partially as a result of the absence of efficient in vitro systems . Here, we report the experimental setup for an Arabidopsis in vitro repair synthesis assay . The repair of plasmid DNA treated with three different DNA-damaging agents, UV light, cisplatin, and methylene blue, after incubation with whole-cell extract was monitored . To validate the reliability of our assay, we analyzed the repair proficiency of plants depleted in AtRAD1 activity . The reduced repair of UV light- and cisplatin-damaged DNA confirmed the deficiency of these plants in nucleotide excision repair . Decreased repair of methylene blue-induced oxidative lesions, which are believed to be processed by the base excision repair machinery in mammalian cells, may indicate a possible involvement of AtRAD1 in the repair of oxidative damage . Differences in sensitivity to DNA polymerase inhibitors (aphidicolin and dideoxy TTP) between plant and human cell extracts were observed with this assay. Annu Rev Physiol, 2002, 64, 223 - 62 Mycosporine-like amino acids and related Gadusols: biosynthesis, acumulation, and UV-protective functions in aquatic organisms; Shick JM et al.; Organisms living in clear, shallow water are exposed to the damaging wavelengths of solar ultraviolet radiation (UVR) coincident with the longer wavelengths of photosynthetically available radiation (PAR) also necessary for vision . With the general exception of bacteria, taxonomically diverse marine and freshwater organisms have evolved the capacity to synthesize or accumulate UV-absorbing mycosporine-like amino acids (MAAs), presumably for protection against environmental UVR . This review highlights the evidence for this UV-protective role while also considering other attributed functions, including reproductive and osmotic regulation and vision . Probing the regulation and biosynthesis of MAAs provides insight to the physiological evolution and utility of UV protection and of biochemically associated antioxidant defenses. J Neurophysiol, 2002 Feb, 87(2), 954 - 61 Ionic basis of the resting membrane potential and action potential in the pharyngeal muscle of Caenorhabditis elegans; Franks CJ et al.; The pharynx of C . elegans is a rhythmically active muscle that pumps bacteria into the gut of the nematode . This activity is maintained by action potentials, which qualitatively bear a resemblance to vertebrate cardiac action potentials . Here, the ionic basis of the resting membrane potential and pharyngeal action potential has been characterized using intracellular recording techniques . The resting membrane potential is largely determined by a K(+) permeability, and a ouabain-sensitive, electrogenic pump . As previously suggested, the action potential is at least partly dependent on voltage-gated Ca(2+) channels, as the amplitude was increased as extracellular Ca(2+) was increased, and decreased by L-type Ca(2+) channel blockers verapamil and nifedipine . Barium caused a marked prolongation of action potential duration, suggesting that a calcium-activated K(+) current may contribute to repolarization . Most notably, however, we found that action potentials were abolished in the absence of external Na(+) . This may be due, at least in part, to a Na(+)-dependent pacemaker potential . In addition, the persistence of action potentials in nominally free Ca(2+), the inhibition by Na(+) channel blockers procaine and quinidine, and the increase in action potential frequency caused by veratridine, a toxin that alters activation of voltage-gated Na(+) channels, point to the involvement of a voltage-gated Na(+) current . Voltage-clamp analysis is required for detailed characterization of this current, and this is in progress . Nonetheless, these observations are quite surprising in view of the lack of any obvious candidate genes for voltage-gated Na(+) channels in the C . elegans genome . It would therefore be informative to re-evaluate the data from these homology searches, with the aim of identifying the gene(s) conferring this Na(+), quinidine, and veratridine sensitivity to the pharynx. J Clin Microbiol, 2002 Feb, 40(2), 637 - 40 Evaluation of Chlamydia pneumoniae and Mycoplasma pneumoniae as etiologic agents of persistent cough in adolescents and adults; Wadowsky RM et al.; Chlamydia pneumoniae and Mycoplasma pneumoniae were evaluated as agents of persistent cough in adolescents and adults (n = 491) . Tests of 473 respiratory specimens by culture or PCR or both identified four episodes (0.8%) of M . pneumoniae-associated illness and no episodes of C . pneumoniae illness, suggesting that these bacteria do not frequently cause persistent cough. Zhonghua Wai Ke Za Zhi, 1998 Dec, 36(12), 756 - 8 {Analysis of clinicopathologic changes of infection in dead patients after renal transplantation}; Xin J et al.; OBJECTIVE: To prevent renal transplantation patients from infection . METHOD: Autopsies of 24 patients who had died after renal transplantation were studied . RESULT: The incidence of infection was 66.7% and the lethal infective complications amounted for 50% . The most infectious agents were bacteria, fungi, tuberculosis, virus and actinomyces . CONCLUSION: Pathological features and clinical manifestations of lesions were caused by aforementioned agents . Some management can prevent renal transplantation recipients from infection. J Gastroenterol, 2002 Jan, 37(1), 10 - 6 Effects of interleukin-8 and Helicobacter pylori on histamine release from isolated canine gastric mucosal mast cells; Yakabi K et al.; BACKGROUND: In recent studies, the involvement of mast cells in the pathogenesis of Helicobacter pylori infection was suggested . In the present study, using isolated canine gastric mucosal mast cells, we undertook to elucidate the effects of interleukin-8 (IL-8) and H . pylori on histamine release from these cells . METHODS: Enriched canine gastric mucosal mast cells (50% target cells) were incubated in Hanks medium with IL-8, or water extract or sonicate of H . pylori for 15 min at 37 degrees C . The content of histamine in the supernatants and the cell pellets after centrifugation was assayed with a histamine radioimmunoassay (RIA) kit . RESULTS: IL-8 (50 ng/ml) and concanavalin A (20 microg/ml) significantly increased histamine release from enriched gastric mucosal mast cells . Dose-dependent stimulation of histamine release by IL-8 (5-50 ng/ml) was also seen . Water extract and sonicate of H . pylori (10(8) bacteria) increased histamine release from mast cells . A concentration-dependent stimulation of histamine release by water extract or sonicate was also seen . The maximal response of histamine release was seen at the highest concentration of the water extract or sonicate . CONCLUSIONS: The results indicated that IL-8 and H . pylori had stimulatory effects on histamine release from canine gastric mucosal mast cells . The results imply that IL-8 and soluble factors of H . pylori may accelerate inflammation of the gastric mucosa via histamine release from mast cells. Facial Plast Surg, 2002 Feb, 18(1), 41 - 52 Growth factor therapy to improve soft tissue healing; Hom DB et al.; The advent of new U.S . Food and Drug Administration-approved products containing growth factors to improve soft tissue healing signifies a new era for wound healing . Over the last decade, some clinical growth factor studies have been promising, whereas others have not shown any positive effect . What many of these studies have shown is that the state of a healing wound is not only dependent on its growth factor milieu but also on other variables (wound care, tissue oxygen state, bacteria count, and nutritional status of the patient) . This article describes recent progress with growth factors on soft tissue structures (skin, mucosa, and nerve) relevant to the facial plastic and reconstructive surgeon. Nature . 2002 Jan 31;415(6871):496. Lysogeny in marine Synechococcus; McDaniel L et al.; Viral infection of bacteria can be lytic, causing destruction of the host cell, or lysogenic, in which the viral genome is instead stably maintained as a prophage within its host . Here we show that lysogeny occurs in natural populations of an autotrophic picoplankton (Synechococcus) and that there is a seasonal pattern to this interaction . Because lysogeny confers immunity to infection by related viruses, this process may account for the resistance to viral infection seen in common forms of autotrophic picoplankton. J Immunol, 2002 Feb 15, 168(4), 1538 - 41 Cutting edge: Tapasin is retained in the endoplasmic reticulum by dynamic clustering and exclusion from endoplasmic reticulum exit sites; Pentcheva T et al.; Tapasin retains empty or suboptimally loaded MHC class I molecules in the endoplasmic reticulum (ER) . However, the molecular mechanism of this process and how tapasin itself is retained in the ER are unknown . These questions were addressed by tagging tapasin with the cyan fluorescent protein or yellow fluorescent protein (YFP) and probing the distribution and mobility of the tagged proteins . YFP-tapasin molecules were functional and could be isolated in association with TAP, as reported for native tapasin . YFP-tapasin was excluded from ER exit sites even after accumulation of secretory cargo due to disrupted anterograde traffic . Almost all tapasin molecules were clustered, and these clusters diffused freely in the ER . Tapasin oligomers appear to be retained by the failure of the export machinery to recognize them as cargo. Appl Environ Microbiol, 2002 Feb, 68(2), 985 - 8 Involvement of glnB, glnZ, and glnD genes in the regulation of poly-3-hydroxybutyrate biosynthesis by ammonia in Azospirillum brasilense Sp7; Sun J et al.; The role of three key nitrogen regulatory genes, glnB (encoding the P(II) protein), glnZ (encoding the P(z) protein), and glnD (encoding the GlnD protein), in regulation of poly-3-hydroxybutyrate (PHB) biosynthesis by ammonia in Azospirillum brasilense Sp7 was investigated . It was observed that glnB glnZ and glnD mutants produce substantially higher amounts of PHB than the wild type produces during the active growth phase . glnB and glnZ mutants have PHB production phenotypes similar to that of the wild type . Our results indicate that the P(II)-P(z) system is apparently involved in nitrogen-dependent regulation of PHB biosynthesis in A . brasilense Sp7. Appl Environ Microbiol, 2002 Feb, 68(2), 968 - 72 Chemotaxis of Ralstonia eutropha JMP134(pJP4) to the herbicide 2,4-dichlorophenoxyacetate; Hawkins AC et al.; Ralstonia eutropha JMP134(pJP4) and several other species of motile bacteria can degrade the herbicide 2,4-dichlorophenoxyacetate (2,4-D), but it was not known if bacteria could sense and swim towards 2,4-D by the process of chemotaxis . Wild-type R . eutropha cells were chemotactically attracted to 2,4-D in swarm plate assays and qualitative capillary assays . The chemotactic response was induced by growth with 2,4-D and depended on the presence of the catabolic plasmid pJP4, which harbors the tfd genes for 2,4-D degradation . The tfd cluster also encodes a permease for 2,4-D named TfdK . A tfdK mutant was not chemotactic to 2,4-D, even though it grew at wild-type rates on 2,4-D. Appl Environ Microbiol, 2002 Feb, 68(2), 852 - 8 Identical ring cleavage products during anaerobic degradation of naphthalene, 2-methylnaphthalene, and tetralin indicate a new metabolic pathway; Annweiler E et al.; Anaerobic degradation of naphthalene, 2-methylnaphthalene, and tetralin (1,2,3,4-tetrahydronaphthalene) was investigated with a sulfate-reducing enrichment culture obtained from a contaminated aquifer . Degradation studies with tetralin revealed 5,6,7,8-tetrahydro-2-naphthoic acid as a major metabolite indicating activation by addition of a C(1) unit to tetralin, comparable to the formation of 2-naphthoic acid in anaerobic naphthalene degradation . The activation reaction was specific for the aromatic ring of tetralin; 1,2,3,4-tetrahydro-2-naphthoic acid was not detected . The reduced 2-naphthoic acid derivatives tetrahydro-, octahydro-, and decahydro-2-naphthoic acid were identified consistently in supernatants of cultures grown with either naphthalene, 2-methylnaphthalene, or tetralin . In addition, two common ring cleavage products were identified . Gas chromatography-mass spectrometry (GC-MS) and high-resolution GC-MS analyses revealed a compound with a cyclohexane ring and two carboxylic acid side chains as one of the first ring cleavage products . The elemental composition was C(11)H(16)O(4) (C(11)H(16)O(4)-diacid), indicating that all carbon atoms of the precursor 2-naphthoic acid structure were preserved in this ring cleavage product . According to the mass spectrum, the side chains could be either an acetic acid and a propenic acid, or a carboxy group and a butenic acid side chain . A further ring cleavage product was identified as 2-carboxycyclohexylacetic acid and was assumed to be formed by beta-oxidation of one of the side chains of the C(11)H(16)O(4)-diacid . Stable isotope-labeling growth experiments with either (13)C-labeled naphthalene, per-deuterated naphthalene-d(8), or a (13)C-bicarbonate-buffered medium showed that the ring cleavage products derived from the introduced carbon source naphthalene . The series of identified metabolites suggests that anaerobic degradation of naphthalenes proceeds via reduction of the aromatic ring system of 2-naphthoic acid to initiate ring cleavage in analogy to the benzoyl-coenzyme A pathway for monoaromatic hydrocarbons . Our findings provide strong indications that further degradation goes through saturated compounds with a cyclohexane ring structure and not through monoaromatic compounds . A metabolic pathway for anaerobic degradation of bicyclic aromatic hydrocarbons with 2-naphthoic acid as the central intermediate is proposed. Appl Environ Microbiol, 2002 Feb, 68(2), 656 - 60 Characterization of Wolbachia host cell range via the in vitro establishment of infections; Dobson SL et al.; Maternally transmitted bacteria of the genus Wolbachia are obligate, intracellular symbionts that are frequently found in insects and cause a diverse array of reproductive manipulations, including cytoplasmic incompatibility, male killing, parthenogenesis, and feminization . Despite the existence of a broad range of scientific interest, many aspects of Wolbachia research have been limited to laboratories with insect-rearing facilities . The inability to culture these bacteria outside of the invertebrate host has also led to the existing bias of Wolbachia research toward infections that occur in host insects that are easily reared . Here, we demonstrate that Wolbachia infections can be simply established, stably maintained, and cryogenically stored in vitro using standard tissue culture techniques . We have examined Wolbachia host range by introducing different Wolbachia types into a single tissue culture . The results show that an Aedes albopictus (Diptera: Culicidae) cell line can support five different Wolbachia infection types derived from Drosophila simulans (Diptera: Drosophilidae), Culex pipiens (Culicidae), and Cadra cautella (Lepidoptera: Phycitidae) . These bacterial types include infection types that have been assigned to two of the major Wolbachia clades . As an additional examination of Wolbachia host cell range, we demonstrated that a Wolbachia strain from D . simulans could be established in host insect cell lines derived from A . albopictus, Spodoptera frugiperda (Lepidoptera: Noctuidae), and Drosophila melanogaster . These results will facilitate the development of a Wolbachia stock center, permitting novel approaches for the study of Wolbachia infections and encouraging Wolbachia research in additional laboratories. Appl Environ Microbiol, 2002 Feb, 68(2), 602 - 7 Effect of commercial-scale high-temperature, short-time pasteurization on the viability of Mycobacterium paratuberculosis in naturally infected cows' milk; Grant IR et al.; Raw cows' milk naturally infected with Mycobacterium paratuberculosis was pasteurized with an APV HXP commercial-scale pasteurizer (capacity 2,000 liters/h) on 12 separate occasions . On each processing occasion, milk was subjected to four different pasteurization treatments, viz., 73 degrees C for 15 s or 25 s with and without prior homogenization (2,500 lb/in(2) in two stages), in an APV Manton Gaulin KF6 homogenizer . Raw and pasteurized milk samples were tested for M . paratuberculosis by immunomagnetic separation (IMS)-PCR (to detect the presence of bacteria) and culture after decontamination with 0.75% (wt/vol) cetylpyridinium chloride for 5 h (to confirm bacterial viability) . On 10 of the 12 processing occasions, M . paratuberculosis was detectable by IMS-PCR, culture, or both in either raw or pasteurized milk . Overall, viable M . paratuberculosis was cultured from 4 (6.7%) of 60 raw and 10 (6.9%) of 144 pasteurized milk samples . On one processing day, in particular, M . paratuberculosis appeared to have been present in greater abundance in the source raw milk (evidenced by more culture positives and stronger PCR signals), and on this occasion, surviving M . paratuberculosis bacteria were isolated from milk processed by all four heat treatments, i.e., 73 degrees C for 15 and 25 s with and without prior homogenization . On one other occasion, surviving M . paratuberculosis bacteria were isolated from an unhomogenized milk sample that had been heat treated at 73 degrees C for 25 s . Results suggested that homogenization increases the lethality of subsequent heat treatment to some extent with respect to M . paratuberculosis, but the extended 25-s holding time at 73 degrees C was found to be no more effective at killing M . paratuberculosis than the standard 15-s holding time . This study provides clear evidence that M . paratuberculosis bacteria in naturally infected milk are capable of surviving commercial high-temperature, short-time pasteurization if they are present in raw milk in sufficient numbers. Appl Environ Microbiol, 2002 Feb, 68(2), 496 - 504 Numerical dominance and phylotype diversity of marine Rhodobacter species during early colonization of submerged surfaces in coastal marine waters as determined by 16S ribosomal DNA sequence analysis and fluorescence in situ hybridization; Dang H et al.; Early stages of surface colonization in coastal marine waters appear to be dominated by the marine Rhodobacter group of the alpha subdivision of the division Proteobacteria (alpha-Proteobacteria) . However, the quantitative contribution of this group to primary surface colonization has not been determined . In this study, glass microscope slides were incubated in a salt marsh tidal creek for 3 or 6 days . Colonizing bacteria on the slides were examined by fluorescence in situ hybridization by employing DNA probes targeting 16S or 23S rRNA to identify specific phylogenetic groups . Confocal laser scanning microscopy was then used to quantify and track the dynamics of bacterial primary colonists during the early stages of surface colonization and growth . More than 60% of the surface-colonizing bacteria detectable by fluorescence staining (Yo-Pro-1) could also be detected with the Bacteria domain probe EUB338 . Archaea were not detected on the surfaces and did not appear to participate in surface colonization . Of the three subdivisions of the Proteobacteria examined, the alpha-Proteobacteria were the most abundant surface-colonizing organisms . More than 28% of the total bacterial cells and more than 40% of the cells detected by EUB338 on the surfaces were affiliated with the marine Rhodobacter group . Bacterial abundance increased significantly on the surfaces during short-term incubation, mainly due to the growth of the marine Rhodobacter group organisms . These results demonstrated the quantitative importance of the marine Rhodobacter group in colonization of surfaces in salt marsh waters and confirmed that at least during the early stages of colonization, this group dominated the surface-colonizing bacterial assemblage. Curr Atheroscler Rep, 2002 Mar, 4(2), 113 - 9 Chronic infections and atherosclerosis/thrombosis; Shah PK; An emerging pathophysiologic paradigm implicates chronic inflammation in the initiation, progression, and destabilization of atherosclerotic vascular disease . Various potential contributors to the inflammatory response in the vessel wall include atherogenic lipids, mechanical stress and injury, hypertension and angiotensin II, cigarette smoking, immune response to neoantigens, and chronic infections with viruses and or bacteria (Table 1) . The potential link between chronic infection and atherosclerosis/thrombosis is under extensive investigation in several laboratories around the world . Although indirect evidence and experimental data tend to support this link, definitive proof is still lacking . If such a link is eventually proven to be causal in nature, it will provide a novel target for preventive and therapeutic strategies (anti-infective drugs, vaccines, etc.) against a common disease that is the leading killer of people in Western nations . Results of ongoing, large-scale clinical trials are eagerly awaited. Syst Appl Microbiol, 2001 Nov, 24(3), 385 - 94 Taxonomic studies of predatory bdellovibrios based on 16S rRNA analysis, ribotyping and the hit locus and characterization of isolates from the gut of animals; Schwudke D et al.; The aim of our study was to obtain data for the molecular characterization of bdellovibrio bacteria, which were recently split into the genus Bdellovibrio and the newly designated genus Bacteriovorax . We determined the 16S rDNA sequences of five reference strains and performed a phylogenetic analysis including published 16S rRNA sequences of bdellovibrios . A comparison of the secondary structure showed significant differences in two regions of the 16S rRNAs of the species Bdellovibrio bacteriovorus, Bacteriovorax starrii, and Bacteriovorax stolpii . In addition, ribotyping techniques gave specific hybridization patterns and revealed that two rRNA operons are present in the investigated strains . A hybridization probe derived from the genetic locus hit, associated with the host independent (HI) phenotype of B . bacteriovorus, was found to be specific for this species . Sequence comparison of the hit locus revealed few base pair changes between host independent (HI) and host dependent (HD) strains . Ribotyping and hybridization experiments using the hit probe were applied to characterize bdellovibrio strains isolated from the gut of animals and humans and one isolate from sewage. Ned Tijdschr Tandheelkd, 1993 Nov, 100(11), 484 - 8 {Amalgam . X . Glass-ionomer cement: a biocompatible substitute for amalgam?}; Schuurs AH et al.; The biocompatibility of glass-ionomer depends upon the components released . The consequences of this releasing for the health of the pulp seem to be moderate, provided that there is a rather thick layer of dentin (> 1 mm) to protect the pulp . Glass-ionomer shrinks upon setting, but it adheres to the dental hard tissues . Yet, bacteria are found along and underneath the material . Systemic effects due to toxicity and allergy are largely unknown, possibly because glass-ionomer is a recent, as yet insufficiently investigated material. Zhonghua Jie He He Hu Xi Za Zhi, 1999 Feb, 22(2), 85 - 7 {Investigation of factors affecting the development of COPD}; Zou X et al.; OBJECTIVE: To investigate the factors which affect the development of COPD . METHODS: 88 patients with mPAP less than 20 mm Hg were followed up by 5 - 20 years . Pulmonary ventilation, blood gas analysis, ECG, pulmonary hemodynamic parameters and oxygen saturation at night were performed one time every 5 years . RESULTS: The results showed that: 5 year, 10 year and 20 year's mortality is 6%, 25% and 43% respectively; MVV%, FEV(1)% and PaO(2) declined accompanied by the increase of PaCO(2); among 41 patients with mPAP 16 - 20 mm Hg, 20 patients' mPAP were more than 20 mm Hg and 15 patients died after 10 years' follow up . Among 16 patients with the declination of oxygen saturation, 8 patients' mPAP were more than 20 mm Hg and 6 patients died . 57 strains of infectious bacteria were found in 88 patients' sputum with 43 strains in the death group . CONCLUSIONS: These results demonstrated that COPD is a kind of chronic diseases with high death rate; infection, decline of oxygen saturation at night and mPAP more than 16 mm Hg were high risk factors in COPD. Biochem Biophys Res Commun, 2002 Feb 8, 290(5), 1545 - 50 Cocaine increases dopamine uptake and cell surface expression of dopamine transporters; Daws LC et al.; In HEK 293 cells expressing the human dopamine transporter (DAT), a 10-min incubation with 10 microM cocaine followed by extensive washing resulted in a 30% increase in {3H}dopamine (DA) uptake as well as an increase in cell surface DAT in biotinylation experiments . Consistent with this novel regulation, {3H}DA uptake into synaptosomes prepared from the nucleus accumbens of rats sacrificed 30 min after a single cocaine injection (30 mg/kg) was significantly increased compared to controls (56% increase in V(max), no change in K(m)) . In addition, DA clearance in the striatum of anesthetized rats was increased after local application of a low (3 pmol) but not high (65 pmol) dose of cocaine, presumably as a result of mobilization of DAT to the cell surface . Cocaine-induced increases in cell surface expression of DAT and associated changes in DA clearance represent a novel mechanism that may play a role in its addictive properties . (c)2002 Elsevier Science (USA). Int Microbiol, 2001 Sep, 4(3), 135 - 42 Enzymatic systems of inorganic pyrophosphate bioenergetics in photosynthetic and heterotrophic protists: remnants or metabolic cornerstones? Perez-Castineira JR, Gomez-Garcia R, Lopez-Marques RL, Losada M, Serrano A. An increasing body of biochemical and genetic evidence suggests that inorganic pyrophosphate (PPi) plays an important role in protist bioenergetics . In these organisms, two types of inorganic pyrophosphatases {EC 3.6.1.1, namely soluble PPases (sPPases) and proton-translocating PPases (H+-PPases)} that hydrolyse the PPi generated by cell anabolism, thereby replenishing the orthophosphate pool needed for phosphorylation reactions, are present in different cellular compartments . Photosynthetic and heterotrophic protists possess sPPases located in cellular organelles (plastids and mitochondria), where many anabolic and biosynthetic reactions take place, in addition to H+-PPases, which are integral membrane proteins of the vacuolysosomal membranes and use the chemical energy of PPi to generate an electrochemical proton gradient useful in cell bioenergetics . This last category of proton pumps was considered to be restricted to higher plants and some primitive photosynthetic bacteria, but it has been found recently in many protists (microalgae and protozoa) and bacteria, thus indicating that H+-PPases are much more widespread than previously thought . No cytosolic sPPase (in bacteria, fungi and animal cells) has been shown to occur in these lower eukaryotes . The widespread occurrence of these key enzymes of PPi metabolism among evolutionarily divergent protists strongly supports the ancestral character of the bioenergetics based on this simple energy-rich compound, which may play an important role in survival under different biotic and abiotic stress conditions. Mycoses, 2001 Dec, 44(11-12), 472 - 9 Otomycosis: prevalence, clinical symptoms, therapeutic procedure; Kurnatowski P et al.; Local lesions observed in otitis, create favourable conditions for the growth of fungi and development of mycoses both in the external and middle ear, as well as in post-operative cavities, especially in cases of open-type surgery . The objective of this study was: (1) to determine the prevalence of mycotic infections in inflammatory conditions of the ear; (2) to evaluate the relationship between the presence of fungi and clinical symptoms of inflammation; and (3) to construct a model of therapeutic procedure . The study concerned 345 individuals, including 295 patients (381 ears) who presented with symptoms of otitis . The mycological examinations revealed presence of fungi in 30.4% cases (116 ears) . Positive culture results were obtained predominantly in patients with post-operative suppuration (52.8%) . Otomycosis was more often associated with fungi of the genus Candida (3/5) than with fungi of the genus Aspergillus (2/5) . The analysis of symptoms in otomycoses showed that in bacteria-induced otitis, pruritus, discharge, post-operative defective hearing and headaches are most common; the analysis of signs showed that reddened epidermis and lining of the tympanic cavity are statistically more frequent . The therapeutic protocol adopted in otomycoses yields good results: 89%--complete cure. Presse Med, 2001 Dec 22-29, 30(39-40 Pt 2), 26 - 32 {Inflammation and seromucous otitis}; Le Clech G; 1 . AN IMPORTANT RELATIONSHIP: There is a strong relationship between tubular dysfunction, tympanic depression and effusion of the tympanic cavity . 2 . TWO IMPORTANT PHENOMENA: Increased secretory capacity of the mucosa and decreased mucociliary clearance subsequent to a reduction in the number of ciliated cells have an important impact . 3 . CONTINUUM: Rhinopharyngeal infection, acute middle ear infection and seromucosal otitis constitutes a continuum demonstrated by the fact that PCR (polymerase chain reaction) studies reveal the same germs in the rhinopharynx, in the middle ear fluid, and in seromucous otitis (SMO) effusions . 4 . BACTERIA AND VIRUSES: By producing toxin, bacteria and viruses induce severe inflammatory reactions in SMO, triggering the cascaded of inflammation mediators . 5 . MEDIATORS: Several elements participating in the cascade of inflammatory events have been demonstrated in human patients and animal models of SMO despite the minimal number of elements producing inflammation mediators initially . Recurrent favoring factors have a primordial effect . 6 . ALLERGY: The clinical and epidemiologic data are widely debated, but do not favor the hypothesis of an allergic reaction, yet several studies have demonstrated that mediators of the allergic response are present in SMO effusions . 7 . ANTIBIOTICS: Used in short-term regimens, antibiotics have a favorable {not readable: see text} on SMO, but their duration of action is short due to the persistent inflammatory reactions . 8 . ORAL CORTICOSTEROIDS: The beneficial effect of oral corticosteroids in SMO is generally short-lived with a rapid decline in efficacy after a few weeks . 9 . LONG DURATION AND RELAPSE: SMO is usually a long-standing highly recurrent condition proscribing use of long-term systemic corticosteroids . It might be interesting to assess the contribution of local corticosteroid therapy after classical "acute" treatment associating antibiotics and a short course of oral corticosteroids . 10 . OTHER OPTIONS: Clinical trials using other treatment options in SMO have not been conclusive for mucolytic, decongestant or antihistamine agents. World J Gastroenterol, 1999 Aug, 5(4), 327 - 329 Intestinal flora translocation and overgrowth in upper gastrointestinal tract induced by hepatic failure; Yi JH et al.; AIM:To explore the relationship between endoinfection caused by intestinal flora translocation and multiple organ dysfunction in hepatic failure.METHODS:By using the quantitative bacteria culture, bacteria colony was counted in GI tract, bile duct and mesenteric lymphonodus in rat hepatic failure model.RESULTS:Intestinal flora migrated up to the upper GI tract and overgrew in stomach and jejunum in rats with hepatic failure.The number of bacteria colonies in the specimens of stomach, jejunum and ileum were 4.7X10(4)/mL, 2.1X10(5)/mL, 5.5X10(6)/mL in experiment group and 4.6X10(2)/mL, 6.1X10(1)/mL, 2.4X10(3)/mL in control group respectively (P < 0.05 ) . Bacteria in bile duct and mesenteric lymphonodus of hepatic failure rats were also cultured . Extensive damages of gastrointestinal mucosa caused by bacterial overgrowth were observed.CONCLUSION:Intestinal flora translocation and overgrowth in stomach and jejunum formed an endoinfectious source and caused obvious pathological injury of gastrointestinal mucosa, which play a very important role in developing abdominal distension, toxic intestinal expansion, alimentary tract haemorrhage and endotoxemia in patients with hepatic failure. Vet Rec, 2002 Jan 5, 150(1), 9 - 11 Induction of enzootic pneumonia in pigs by the administration of an aerosol of in vitro-cultured Mycoplasma hyopneumoniae; Czaja T et al.; The aim of this study was to establish whether enzootic pneumonia could be induced reliably in piglets by administering an aerosolised culture of Mycoplasma hyopneumoniae . Groups of five M hyopneumoniaefree Landrace x Large White piglets weaned at 11 to 14 days of age were exposed to aerosols of in vitro cultures of a virulent strain of M hyopneumoniae . In three separate trials, 14 of 15 pigs exposed to the bacteria developed pneumonia, but pigs exposed to the culture medium alone did not develop the disease . Lung pathology, both gross and histological, indicated acute disease . Ten of the pigs were tested for seroconversion by Western blot and they were all positive . The growth rates of the infected pigs were significantly reduced and the water consumption of the infected groups was also depressed . M hyopneumoniae was recovered from eight of the 15 infected pigs. Immunol Res, 2001, 24(3), 225 - 44 Nucleic acid vaccines: tasks and tactics; McKenzie BS et al.; There are no adequate vaccines against some of the new or reemerged infectious scourges such as HIV and TB . They may require strong and enduring cell-mediated immunity to be elicited . This is quite a task, as the only known basis of protection by current commercial vaccines is antibody . As DNA or RNA vaccines may induce both cell-mediated and humoral immunity, great interest has been shown in them . However, doubt remains whether their efficacy will suffice for their clinical realization . We look at the various tactics to increase the potency of nucleic acid vaccines and divided them broadly under those affecting delivery and those affecting immune induction . For delivery, we have considered ways of improving uptake and the use of bacterial, replicon or viral vectors . For immune induction, we considered aspects of immunostimulatory CpG motifs, coinjection of cytokines or costimulators and alterations of the antigen, its cellular localization and its anatomical localization including the use of ligand-targeting to lymphoid tissue . We also thought that mucosal application of DNA deserved a separate section . In this review, we have taken the liberty to discuss these enhancement methods, whenever possible, in the context of the underlying mechanisms that might argue for or against these strategies. Antonie Van Leeuwenhoek, 2001 Sep, 79(3-4), 385 - 91 The root-nodule symbiosis between Sarothamnus scoparius L . and its microsymbionts; Sajnaga E et al.; When nitrogen fixing root nodules are formed, Sarothamnus scoparius (broom) is inoculated with its microsymbionts . Nodules studied under light and electron microscopy exhibited typical indeterminate nodule histology with apical, persistent meristem, age gradient of nodule tissues, and open vascular bundles, and also with some particular features such as: the presence of mitotic activity in the infected meristematic cells, lack of infection threads, distribution of bacteria by process of host cell division, and occurrence of a large bacteroid zone only with infected cells . The results of cross-inoculation tests have shown a broad host range for S . scoparius microsymbionts including not only the native host but also species such as: Lupinus luteus, Ornithopus sativa, Lotus corniculatus, Genista tinctoria, Chamaecitisus ratisbonensis, Macroptilium atropurpureum, and Phaseolus vulgaris . In addition, our data established a close symbiotic relationship of S . scoparius nodulators to Bradyrhizobium sp . (Lupinus) by comparison of the partial sequence of nodC gene of the strain CYT7, specific for the broom, to those from Bradyrhizobium sp . (Lupinus) strain D1 and others available in the public databases. J Cell Biol, 2002 Feb 4, 156(3), 419 - 24 Epub 2002 Jan 28. Condensin and cohesin display different arm conformations with characteristic hinge angles; Anderson DE et al.; Structural maintenance of chromosomes (SMC) proteins play central roles in higher-order chromosome dynamics from bacteria to humans . In eukaryotes, two different SMC protein complexes, condensin and cohesin, regulate chromosome condensation and sister chromatid cohesion, respectively . Each of the complexes consists of a heterodimeric pair of SMC subunits and two or three non-SMC subunits . Previous studies have shown that a bacterial SMC homodimer has a symmetrical structure in which two long coiled-coil arms are connected by a flexible hinge . A catalytic domain with DNA- and ATP-binding activities is located at the distal end of each arm . We report here the visualization of vertebrate condensin and cohesin by electron microscopy . Both complexes display the two-armed structure characteristic of SMC proteins, but their conformations are remarkably different . The hinge of condensin is closed and the coiled-coil arms are placed close together . In contrast, the hinge of cohesin is wide open and the coiled-coils are spread apart from each other . The non-SMC subunits of both condensin and cohesin form a globular complex bound to the catalytic domains of the SMC heterodimers . We propose that the "closed" conformation of condensin and the "open" conformation of cohesin are important structural properties that contribute to their specialized biochemical and physiological functions. FEMS Microbiol Lett, 2002 Jan 10, 206(2), 215 - 9 Transformation of the cyanobacterium Synechocystis sp . PCC 6803 as a tool for genetic mapping: optimization of efficiency; Kufryk GI et al.; The cyanobacterium Synechocystis sp . PCC 6803 is transformable at high efficiency and integrates DNA by homologous double recombination . However, several genetic mapping procedures depend on the ability to generate transformants even with very small amounts of added DNA . This study is aimed at optimizing the transformation efficiency at limiting concentrations of exogenous DNA . The transformation efficiency showed little sensitivity to experimental conditions . Transformation with circular plasmid DNA was found to be no more than 30% more efficient than with linearized plasmid DNA . The efficiency of transformation remained essentially the same in the presence of competing DNA, indicating that the capacity of DNA uptake by the cells is not limiting . The incubation time of cells with DNA before plating (0-8 h) affected the transformation efficiency by up to 3-fold . Only minor changes in the efficiency were observed as a function of the presence of a membrane filter on the plate or the presence of TAE or TBE gel buffer residues in the transformation mixture . However, transformability of the host strain of Synechocystis sp . PCC 6803 was increased by two orders of magnitude if the sll1354 gene encoding the exonuclease RecJ was deleted . Therefore, the transformation efficiency of Synechocystis sp . PCC 6803 with exogenous DNA appears to be determined primarily by intracellular processes such as the efficiency of DNA processing and homologous recombination. Trends Biotechnol, 2002 Feb, 20(2), 65 - 71 Recombinant immunoglobulin A: powerful tools for fundamental and applied research; Corthesy B; The use of monoclonal antibodies has become routine in research and diagnostic laboratories but the potential level of antibodies in use in public health and medical applications is still far from its maximum . From a clinical perspective, topical immunotherapy of mucosal surfaces with monoclonal antibodies can block entry and transmission of bacteria, viruses, fungi and parasites that infect humans, and defeat some key strategies, evolved by many pathogens, to evade the host immune system . The chief antibody at mucosal surfaces is secretory immunoglobulin A (SIgA), a multi-polypeptide complex originating from two cell types . The recent design of heterologous expression systems, coupled with modern biotechnology processes, should form a sound basis for studying the functional properties of SIgAs and evaluate their value as biotherapeutics . Here, we discuss the principles underlying mucosal immunity and review the application of recombinant SIgA to the dissection of mechanisms in passive and active protection at mucosal surfaces. J Comp Pathol, 2002 Jan, 126(1), 9 - 16 Thermolysin activates equine lamellar hoof matrix metalloproteinases; Mungall BA et al.; Cultured equine lamellar hoof explants secrete the pro-enzymes matrix metalloproteinase-2 (MMP-2, 72 kDa) and MMP-2 (92 kDa) . Untreated explants remained intact when tested on a calibrated force transducer, but when treated with an MMP activator, developed "in-vitro laminitis", separating at the dermal-epidermal junction . Explants treated with the bacterial protease thermolysin separated dose-dependently; this was accompanied by activation of both MMP-2 and -9 . Thermolysin-mediated MP activation did not occur in a cell-free system and was not inhibited by the addition of the MMP inhibitor and batimastat . These findings suggest that thermolysin-mediated gelatinase activation is not dependent on membrane-bound matrix metalloproteinase (MT-MMP) activation, providing further evidence that bacteria can produce potent MMP activators that probably facilitate host invasion. Hepatogastroenterology, 2001 Nov-Dec, 48(42), 1548 - 51 Helicobacter pylori infection and precancerous lesions of the stomach; Valle J et al.; H . pylori infection is associated with a slightly increased risk of gastric cancer . However, the risk is much higher in the subgroup of infected patients who have atrophic gastritis and extensive intestinal metaplasia . In those subjects, H . pylori acts as a trigger of the sequence which begins as atrophic gastritis with intestinal metaplasia and evolves towards immature forms of intestinal metaplasia and dysplasia . It seems that factors different from H . pylori (diet, genetical background, etc.) might have an influence on how often gastric precancerous lesions appear in H . pylori-infected subjects . Effective prevention of gastric carcinoma would require identification of the patients at risk in an early step of the process . Preventive measures would include H . pylori eradication and changes in the diet (i.e., increase of vitamin C and carotenoid intake) . Preliminary data suggest that H . pylori eradication cannot revert intestinal metaplasia . However, it is possible that eradication of the bacteria would prevent progression towards immature forms of intestinal metaplasia and dysplasia. Ying Yong Sheng Tai Xue Bao, 2001 Feb, 12(1), 121 - 5 {Effect of fly ash-filtered mud mixture on soil properties and radish yield and quality}; Xing S et al.; Based on pot culture experiment, the effect of fly ash-filtered mud mixture on soil biochemical properties, radish yield and its quality, and heavy metal accumulation in both soil and radish was examined . The mixture was made by mixing fly ash and filtered mud in proportion 1:1(w/w) and adding small amount of inorganic fertilizer . Its Cd, Pb, Cr, As and Hg contents were much lower than those requested by state control criteria . After applying this mixture in a definite amount, no significant accumulation of heavy metals was found in both soil and radish, the pollution index of heavy metals was < 1, the quantities of soil bacteria increased notably, and the activities of soil urease, phosphates and cellulosase also raised significantly . The application of the mixture promoted radish growth and its development and metabolism . The reducing sugar and vitamin C in radish also increased markedly . The results indicated that the use of adequate amounts of the mixture did not cause any obvious heavy metal pollution in both soil and radish, but could improve soil fertility, and raise radish yield and its quality remarkably. Nucleic Acids Res . 2001 Dec 15;29(24):E120. A unique type II restriction endonuclease FspAI, that recognizes the octanucleotide sequence 5'-RTGC/GCAY-3'; Kesminiene A et al.; A new type II restriction endonuclease designated FspAI has been partially purified from a Flexibacter species Tv-m21K . FspAI recognizes the octanucleotide sequence 5'-RTGC/GCAY-3' and cleaves it in the center generating blunt-ended DNA fragments. Nucleic Acids Res, 2001 Dec 15, 29(24), 5017 - 28 Distribution of substitution rates and location of insertion sites in the tertiary structure of ribosomal RNA; Wuyts J et al.; The relative substitution rate of each nucleotide site in bacterial small subunit rRNA, large subunit rRNA and 5S rRNA was calculated from sequence alignments for each molecule . Two-dimensional and three-dimensional variability maps of the rRNAs were obtained by plotting the substitution rates on secondary structure models and on the tertiary structure of the rRNAs available from X-ray diffraction results . This showed that the substitution rates are generally low near the centre of the ribosome, where the nucleotides essential for its function are situated, and that they increase towards the surface . An inventory was made of insertions characteristic of the Archaea, Bacteria and Eucarya domains, and for additional insertions present in specific eukaryotic taxa . All these insertions occur at the ribosome surface . The taxon-specific insertions seem to arise randomly in the eukaryotic evolutionary tree, without any phylogenetic relatedness between the taxa possessing them. Epidemiol Infect, 2001 Dec, 127(3), 389 - 97 A comparison of gastroenteritis in a general practice-based study and a community-based study; de Wit MA et al.; We compared gastroenteritis cases that consulted a general practitioner (GP) with those who did not in a community-based study and also with those in a GP-based study . We aimed to identify factors associated with consultation, and with inclusion of cases by GPs, and secondly to study the effects on the frequency of detection of pathogens . Furthermore, we estimated the under-ascertainment by GPs . Both studies were performed in The Netherlands in the same population in an overlapping time-period . Overall, 5% of community cases consulted a GP . Cases who consulted suffered from more severe episodes than non-consulting cases . Inclusion of cases by GPs, instead of a study team, caused a selection of more severe cases with more chronic symptoms . When extrapolating data from GP-based studies, it should be taken into account that, in general practice, gastroenteritis due to bacteria and Giardia lamblia is a relatively large proportion of that in the community and gastroenteritis due to Norwalk-like viruses is a relatively small proportion . The incidence of gastroenteritis in general practices was estimated between 14 and 35 per 1000 person years. J Periodontol, 2001 Dec, 72(12), 1742 - 6 Distribution of the receptor for advanced glycation end products gene polymorphisms in patients with chronic periodontitis: a preliminary study; Holla LI et al.; BACKGROUND: Periodontal diseases are viewed today as multifactorial problems initiated and sustained by bacteria but significantly modified by the body's response to bacterial plaque . A recent study suggested that receptor for advanced glycation end products (RAGE) could be involved in the pathophysiology of periodontitis . The aim of this study was to investigate a possible association of 3 common polymorphisms in the RAGE gene with chronic periodontitis . METHODS: We studied 101 Caucasian patients with chronic periodontitis together with 162 orally healthy subjects . Three polymorphisms, one in intron 7 (1704G/T), second in intron 8 (2184A/G), and the third in exon 3 (G82S) of the RAGE gene, were investigated by polymerase chain reaction methods (PCR) with subsequent enzymatic restriction with Bfal, BsmFI, or Alu Il, respectively . RESULTS: A statistically significant difference in allele frequencies between patients and the reference group was found for intron variant 1704G/T (P= 0.02, Pcorr >0.05) . There was no significant difference in genotype or allele frequency distributions between groups for intron variant 2184A/G or for the exon variant exchanging amino acid Gly for Ser at position 82 (G82S) . CONCLUSIONS: We can speculate that susceptibility to the development of chronic periodontitis could be influenced by the 1704G/T polymorphism of the RAGE gene, independently of diabetes. Coll Antropol, 2001 Dec, 25(2), 703 - 12 Current concept on the anticaries fluoride mechanism of the action; Rosin-Grget K et al.; The paper discusses a possible new concept of the role of fluoride and its mechanism of action in caries prevention . In the past fluoride inhibition of caries was ascribed to reduced solubility due to incorporation of fluoride (F) into the enamel minerals (firmly bound fluoride or fluorapatite) . Based on the new findings, it appears that fluoride, either released into or present in the fluid phase bathing the hard tissue, is more important for the reduction of caries development and progression . There is convincing evidence that fluoride has a major effect on demineralization and remineralization of dental hard tissue and that it interferes with acid production from cariogenic bacteria . The provision of dissolved fluoride is the key to successful therapy . The source of this fluoride could either be fluorapatite or calcium fluoride (CaF2) (like) precipitates, which are formed on the enamel and in the plaque after application of topical fluoride . The precipitates of calcium fluoride do not dissolve quickly as was initially believed . Calcium fluoride coating at neutral pH by pellicle proteins and phosphate is the main reason for this . The dissolution of the fluoride from calcium fluoride is pH dependent . At lower pH, the coating is lost and an increased dissolution rate of calcium fluoride occurs . The CaF2, therefore, act as an efficient source of free fluoride ions during the cariogenic challenge . These are subsequently incorporated into the enamel as hydroxyfluorapatite or fluorapatite. J Infect Chemother, 2001 Sep, 7(3), 148 - 55 Allelic exchange mutagenesis of rpoN encoding RNA-polymerase sigma54 subunit in Helicobacter pylori; Fujinaga R et al.; The rpoN gene, encoding the alternative sigma factor (sigma54) of Helicobacter pylori, was amplified from genomic DNA . H . pylori rpoN has an overall similarity to the rpoN of other bacteria, but lacks a glutamine (Q)-rich region in region I and an acidic region in region II . When the rpoN gene was disrupted, the mutant was found to be completely nonmotile . Because the flaB gene has an rpoN consensus sequence in its promoter region, we assessed the transcriptional activity of the flaB gene, using xylE transcriptional fusion . In the isogenic mutant of rpoN, transcription of the flaB gene was severely affected, but transcription of the ureA gene (control) was intact . In late stationary phase, the rpoN mutant showed marked decreases in viability: i.e., the number of colony-forming units (CFU) at 100 h was 4 log lower in the rpoN mutant than in the wild-type strain . By morphological examination with acridine orange staining, the rpoN mutant showed green and faintly orange-stained irregularly shaped cells with a few orange-stained rod/spiral cells . In contrast, the wild-type strain and the non-flagella flgE mutant (control) contained many orange-stained rod/spiral and coccoid cells . These results indicated that in H . pylori, RpoN is involved not only in motility but also in viability, through the morphological changes in the stationary phase. Mol Genet Genomics, 2002 Jan, 266(5), 813 - 20 Epub 2001 Oct 30. Cloning and characterisation of the Azospirillum brasilense glnD gene and analysis of a glnD mutant; Van Dommelen A et al.; Nitrogen regulation in bacteria involves the capacity to sense the availability of fixed nitrogen and to translate a signal indicating nitrogen deficiency or nitrogen excess into a cellular response . One of the key enzymes in this complex regulation process, the uridylyltransferase/uridylyl-removing (UTase) enzyme, encoded by the glnD gene, was characterised in the diazotroph Azospirillum brasilense, which promotes plant growth . The glnD gene product is responsible for the uridylylation of both P(II)-like nitrogen regulatory proteins, P(II) and P(Z), depending on the nitrogen status of the cell . The nitrogen-regulated activity of the main ammonium-assimilating enzyme, glutamine synthetase, is not altered in a glnD-Tn 5-B30 insertion mutant . UTase influences processes that are regulated by the NtrB-NtrC two-component histidine protein kinase system, such as ammonium uptake and nitrate assimilation . Moreover, the glnD gene product is indispensable for the activation of nitrogen fixation . Transcription of glnD is up-regulated under nitrogen-fixing conditions . This regulation is only partially dependent on the global nitrogen regulation (Ntr) system. Science, 2002 Jan 25, 295(5555), 641 - 4 Antarctic Sea ice--a habitat for extremophiles; Thomas DN et al.; The pack ice of Earth's polar oceans appears to be frozen white desert, devoid of life . However, beneath the snow lies a unique habitat for a group of bacteria and microscopic plants and animals that are encased in an ice matrix at low temperatures and light levels, with the only liquid being pockets of concentrated brines . Survival in these conditions requires a complex suite of physiological and metabolic adaptations, but sea-ice organisms thrive in the ice, and their prolific growth ensures they play a fundamental role in polar ecosystems . Apart from their ecological importance, the bacterial and algae species found in sea ice have become the focus for novel biotechnology, as well as being considered proxies for possible life forms on ice-covered extraterrestrial bodies. Mol Cell Biol, 2002 Feb, 22(4), 979 - 91 Role for the adaptor protein Grb10 in the activation of Akt; Jahn T et al.; Grb10 is a member of the Grb7 family of adapter proteins lacking intrinsic enzymatic function and encodes functional domains including a pleckstrin homology (PH) domain and an SH2 domain . The role of different Grb10 splice variants in signal transduction of growth factors like insulin or insulin-like growth factor has been described as inhibitory or stimulatory depending on the presence of a functional PH and/or SH2 domain . Performing a yeast two-hybrid screen with the c-kit cytoplasmic tail fused to LexA as a bait and a mouse embryo cDNA library as prey, we found that the Grb10 SH2 domain interacted with the c-kit receptor tyrosine kinase . In the course of SCF-mediated activation of c-kit, Grb10 is recruited to the c-kit receptor in an SH2 domain- and phosphotyrosine-dependent but PH domain-independent manner . We found that Akt and Grb10 form a constitutive complex, suggesting a role for Grb10 in the translocation of Akt to the cell membrane . Indeed, coexpression studies revealed that Grb10 and c-kit activate Akt in a synergistic manner . This dose-dependent effect of Grb10 is wortmannin sensitive and was also seen at a lower level in cells in which c-kit was not expressed . Expression of a Grb10 mutant lacking the SH2 domain as well as a mutant lacking the PH domain did not influence Akt activity . Grb10-induced Akt activation was observed without increased phosphatidylinositol 3-kinase (PI3-kinase) activity, suggesting that Grb10 is a positive regulator of Akt downstream of PI3-kinase . Significantly, deficient activation of Akt by a constitutively activated c-kit mutant lacking the binding site for PI3-kinase (c-kitD814V/Y719F) could be fully compensated by overexpression of Grb10 . In Ba/F3 cells, the incapacity of c-kitD814V/Y719F to induce interleukin-3 (IL-3)-independent growth could be rescued by overexpression of Grb10 . In contrast, expression of the SH2 deletion mutant of Grb10 together with c-kitD814V/Y719F did not render Ba/F3 cells independent of IL-3 . In summary, we provide evidence that Grb10 is part of the c-kit signaling pathway and that the expression level of Grb10 critically influences Akt activity . We propose a model in which Grb10 acts as a coactivator for Akt by virtue of its ability to form a complex with Akt and its SH2 domain-dependent translocation to the cell membrane. Int J Biochem Cell Biol, 2002 Feb, 34(2), 109 - 15 Cloning and characterization of a novel human pantothenate kinase gene; Ni X et al.; Pantothenate kinase (PanK) is a key regulatory enzyme in the CoA biosynthetic pathway in bacteria and mammalian cells . It catalyzes the first committed step in the universal biosynthetic pathway leading to CoA . Here we report the molecular cloning and characterization of a new human PanK gene . The gene encoded a protein of 314 amino acid residues, which share high homology to mouse pantothenate kinase (mPanK) 1 beta . Northern blot analysis revealed a transcript of the gene of 2.6 kb in human liver and kidney . The human PanK gene was located to human chromosome between 10q22.3 and 10q23.2 by bioinformatics analysis. Lancet, 2002 Jan 5, 359(9300), 62 - 9 Crohn's disease; Shanahan F; Crohn's disease is a disorder mediated by T lymphocytes which arises in genetically susceptible individuals as a result of a breakdown in the regulatory constraints on mucosal immune responses to enteric bacteria . Regulation of immune reactivity to enteric antigens has improved understanding of the pathophysiological mechanisms of Crohn's disease, and has expanded therapeutic options for patients with this disorder . Disease heterogeneity is probable, with various underlying defects associated with a similar pathophysiological outcome . Although most conventional drug treatments are directed at modification of host response, therapeutic manipulation of the enteric flora is becoming a realistic option. Biotechniques, 2002 Jan, 32(1), 134 - 6, 138, 140-1 Synchrotron radiation-induced X-ray emission to identify metal ions in preparations of purified protein; Pfister T et al.; The suitability of synchrotron radiation-induced X-ray emission (SRIXE) for the detection and identification of metal ions in preparations of purified, soluble proteins was tested . Glutathione S-transferase fused to the proximal zinc finger motif of human transcription factor IIIA or to the cysteine-rich motif of poliovirus protein 2C was expressed in bacteria and purified by affinity chromatography . Aqueous samples containing the purified proteins were analyzed with SRIXE, and trace amounts of zinc and iron were detected . Mutation of the zinc-coordinating residues in the cysteine-rich motif of poliovirus protein 2C resulted in the loss of the zinc-binding ability . Relative quantities of metal in the protein preparations as determined by SRIXE corresponded well with the metal:protein ratios calculated by using a 4-(2-pyridylazo)resorcinol-based assay . We conclude that SRIXE is an accurate, sensitive, and simple method for the detection and identification of protein-bound metal ions in small amounts of sample . Thus, SRIXE may have wide use as a particularly effective method for rapidly determining trace metals in microarray samples. Medicina (B Aires), 2001, 61(6), 849 - 51 {Myopericarditis due to enterovirus in association with rhabdomyolysis and renal failure}; Cacace Linares N et al.; We present the case of a 49 year old man who was admitted with odynophagia, fever and abdominal pain . Later he developed dyspnea and polymialgias . Pericardial effusion was detected on the echocardiogram . Renal failure and rhabdomyolysis developed worsening the clinical picture . A pericardial surgical drainage was decided due to cardiac tamponade . All samples were negative for bacteria and fungi . The presence of enterovirus in pericardial fluid was confirmed. J Infect Dis, 2002 Feb 1, 185(3), 401 - 4 Epub 2002 Jan 17. Molecular evidence of endogenous reactivation of Mycobacterium tuberculosis after 33 years of latent infection; Lillebaek T et al.; Since Robert Koch described the cause of tuberculosis in 1882, the natural history of the disease after primary infection has been subject to debate . Only approximately 10% of infected individuals develop active disease, which may appear years to decades after infection . Late onset has been attributed to the endogenous reactivation of dormant bacteria . However, this has not been documented by molecular means for latencies of more than a few years . In Denmark, we have recently recultured 205 freeze-dried Mycobacterium tuberculosis strains obtained from 1961 through 1967 . These "historical" strains are analyzed by DNA restriction fragment-length polymorphism testing, and their DNA patterns are compared with those of 4008 recently obtained clinical specimens . This has, surprisingly, yielded molecular evidence of M . tuberculosis reactivation after 33 years of latent infection . A father and son who developed tuberculosis in 1961 and in 1994, respectively, were the only patients infected with strains that share an identical DNA pattern. J Infect Dis, 2002 Feb 1, 185(3), 341 - 7 Epub 2002 Jan 17. The evaluation of putative virulence factors of Helicobacter pylori for gastroduodenal disease by use of a short-term Mongolian gerbil infection model; Akanuma M et al.; Few virulence determinants of Helicobacter pylori have been tested in vivo . We conducted this study to establish an animal model for their screening . Six-week-old male Mongolian gerbils were inoculated with wild-type H . pylori (TN2) or its isogenic mutant with deletion of cagE (TN2DeltacagE), total cag pathogenicity island (TN2DeltacagPAI), HP0499 (TN2DeltaHP499), or HP0638 (TN2DeltaHP638) (n=5 each) . The animals were killed 3 weeks later, and the density of bacteria and the degree of inflammation in the stomach were compared . Infection was established in all animals except those inoculated with TN2DeltaHP638 . TN2 and TN2DeltaHP499, but not TN2DeltacagE and TN2Deltacag PAI, induced intense inflammation, although the densities of bacteria were similar . The Mongolian gerbil model was useful for the screening of virulence determinants in vivo, which confirmed the importance of cag PAI while questioning that of HP0499. J Bacteriol, 2002 Feb, 184(4), 1214 - 8 Construction and synchronization of dnaA temperature-sensitive mutants of Streptomyces; Lee LF et al.; Temperature-sensitive mutants of Streptomyces defective in initiation of chromosome replication were created by in vitro site-directed mutagenesis in the dnaA gene followed by gene replacement . When they were shifted to 39 degrees C replication in the mutants ceased in about 90 min but resumed on return to 30 degrees C . This allowed manipulations to achieve replication synchronization. Kansenshogaku Zasshi, 2001 Dec, 75(12), 1057 - 61 {A case of leptospirosis infected in Borneo Island, Malaysia}; Sakamoto M et al.; We report a case of leptospirosis infected in Sabah, Borneo island, Malaysia . The case is 25-year-old male who had participated in the EcoChallenge Sabah 2000 Expedition Race, a multisport event held during August 20 to September 3, 2000 at various sites in Sabah in Malaysian Borneo . He developed a high fever and headache on September 7, and he was admitted to our hospital on September 9 . On admission he also had conjunctivitis and myalgias . Laboratory findings on admission revealed leukocytosis with left shift, slightly elevated transaminase levels, high CRP levels and proteinuria . Plasmodium spp . were negative on blood smears, and no bacteria were isolated from blood and feces cultures . We performed the laboratory tests for leptospirosis, based on the information about the probable leptospirosis outbreak among athletes who participated in the EcoChallenge Race, however both Leptospira antigens and antibodies were negative at that time . We diagnosed leptospirosis clinically because he manifested persistent symptoms, and minocycline 100 mg b.i.d . was administered intravenously resulting in excellent efficacy . Serum antibody tests by microscopic agglutination test (MAT) at convalescent stage revealed significant increased antibodies against Leptospira interrogans serovar hebdomadis, and the diagnosis of leptospirosis was confirmed . Infectious diseases have been global and it is important to have information concerning worldwide infectious disease situations as much as possible for accurate diagnosis. Anal Chem, 2002 Jan 1, 74(1), 91 - 5 Determination of the carbon content of airborne fungal spores; Bauer H et al.; Airborne fungal spores contribute potentially to the organic carbon of the atmospheric aerosol, mainly in the "coarse aerosol" size range 2.5-10 microm aerodynamic equivalent diameter (aed) . Here, we report about a procedure to determine the organic carbon content of fungal spores frequently observed in the atmosphere . Furthermore, we apply a new (carbon/individual) factor to quantify the amount of fungal-spores-derived organic carbon in aerosol collected at a mountain site in Austria . Spores of representatives of Cladosporium sp., Aspergillus sp., Penicillium sp., and Alternaria sp., the four predominant airborne genera, were analyzed for their carbon content using two different analytical procedures . The result was an average carbon content of 13 pg C/spore (RSD, 46%), or expressed as a carbon-per-volume ratio, 0.38 pg C/microm3 (RSD, 30%) . These values are comparable to conversion factors for bacteria and some representatives of the zooplankton . Because biopolymers are suspected of interfering with elemental carbon determination by thermal methods, the amount of "fungal carbon" that might be erroneously mistaken for soot carbon was determined using the "two-step combustion" method of Cachier et al . and termed as "apparent elemental carbon" (AEC) . This fraction amounted to up to 46% of the initial fungal carbon content . Although the aerosol samples were collected in March under wintry conditions, the organic carbon from fungal spores amounted to 2.9-5.4% of organic carbon in the "coarse mode" size fraction. Anal Chem, 2002 Jan 1, 74(1), 163 - 70 Direct voltammetry and catalysis with Mycobacterium tuberculosis catalase-peroxidase, peroxidases, and catalase in lipid films; Zhang Z et al.; Stable films of dimyristoylphosphatidylcholine and M . tuberculosis catalase-peroxidase (KatG), several peroxidases, myoglobin, and catalase showed reversible FeIII/FeII voltammetry on pyrolytic graphite electrodes and catalytic current for hydrogen peroxide and oxygen . Amperometric responses for these films to H2O2 at 0 V are likely to contain significant contributions from catalytic reduction of oxygen produced during the catalytic cycles . Relative apparent turnover rates at pH 6 based on steady-state currents at 0 V versus SCE in the presence of H2O2 were in the order horseradish peroxidase > cytochrome c peroxidase (CcP) > soybean peroxidase > myoglobin > KatG > catalase . Lower currents for the very efficient peroxide scavengers KatG and catalase may be related to the instability of their compounds I in the presence of H2O2 . KatG catalyzed the electrochemical reduction of oxygen more efficiently than catalase and CcP but less efficiently than the other peroxidases . DMPC films incorporating glucose oxidase and peroxidases gave good analytical responses to glucose, demonstrating the feasibility of dual enzyme-lipid films for biosensor fabrication. Anal Chem, 2002 Jan 1, 74(1), 114 - 9 Scanning electrochemical microscopy of living cells . 3 . Rhodobacter sphaeroides; Cai C et al.; The scanning electrochemical microscope (SECM) was used to probe the redox activity of individual purple bacteria (Rhodobacter sphaeroides) . The approaches developed in our previous studies of mammalian cells were expanded to measure the rates and investigate the pathway of transmembrane charge transfer in bacteria . The two groups of redox mediators (i.e., hydrophilic and hydrophobic redox species) were used to shuttle the electrons between the SECM tip electrode in solution and the redox centers inside the cell . The analysis of the dependencies of the measured rate constant on formal potential and concentration of mediator species in solution yielded information about the permeability of the outer cell membrane to different ionic species and intracellular redox properties . The maps of redox reactivity of the cell surface were obtained with a micrometer or submicrometer spatial resolution. Ann N Y Acad Sci, 2001 Dec, 953, 146 - 50 Letting the genome out of the bottle: prospects for new drug development; Young D; Use of the information gained from sequencing the Mycobacterium tuberculosis genome will enable scientists to accelerate the development of reagents for improved tuberculosis control . Cloning and expressing genes encoding the enzymes involved in cell-wall biosynthesis will provide the tools for screening millions of novel compounds . Cell wall inhibitors will be mainly useful in treating resistant disease, but cost factors are likely to limit the application of novel compounds in the design of new treatment regimens . More effective might be an approach to target metabolic processes that are essential even in nondividing bacteria . A third target for drug action is elimination of latent disease through a drug that acts in synergy with the immune response. Postepy Hig Med Dosw, 2001, 55(5), 733 - 54 {Non mucin proteins of saliva with high homology of polypeptide chains}; Zalewska A et al.; To non mucin proteins of human saliva belong: cystatins, statherin, histatins and acidic proline-rich protein . These saliva proteins influence hard and soft tissues by forming a pellicle layer on oral mucosa and enamel, by taking part in removing bacteria or initiating of bacterial colonization . Most of them are able to inhibit the formation of dental calculus and control the calcium phosphate homeostasis. Mol Cell, 2002 Jan, 9(1), 195 - 201 ResT, a telomere resolvase encoded by the Lyme disease spirochete; Kobryn K et al.; The genus Borrelia includes the causative agents of Lyme disease and relapsing fever . An unusual feature of these bacteria is a segmented genome consisting mostly of a number of linear DNA molecules with covalently closed hairpin ends or telomeres . In this study we show that the BBB03 locus encodes the B . burgdorferi telomere resolvase, ResT . The purified protein catalyzes telomere resolution in vitro through a unique reaction: breakage of two phosphodiester bonds in a single DNA duplex (one on each strand) and joining of each end with the opposite DNA strand to form covalently closed hairpin telomeres . Telomere resolution by ResT occurs through a two-step transesterification reaction involving the formation of a covalent protein-DNA intermediate at a position three nucleotides from the axis of symmetry in each strand of the substrate. Dis Aquat Organ, 2001 Dec 5, 47(3), 209 - 17 A histological study of shell disease syndrome in the edible crab Cancer pagurus; Vogan CL et al.; Shell disease syndrome is characterised by the external manifestation of black spot lesions in the exoskeletons of crustaceans . In the present study, gills, hepatopancreas and hearts from healthy (<0.05% black spot coverage) and diseased (5 to 15% coverage) edible crabs, Cancer pagurus, were examined histologically to determine whether this disease can cause internal damage to such crabs . There was clear evidence of cuticular damage in the gills of diseased crabs leading to the formation of haemocyte plugs termed nodules . Nephrocytes found within the branchial septa of the gills showed an increase in the accumulation of dark material in their vacuoles in response to disease . In the hepatopancreas, various stages of tubular degradation were apparent that correlated with the severity of external disease . Similarly, there was a positive correlation between the number of viable bacteria in the haemolymph and the degree of shell disease severity . Approximately 21% of the haemolymph-isolated bacteria displayed chitinolytic activity . Overall, these findings suggest that shell disease syndrome should not be considered as a disease of the cuticle alone . Furthermore, it shows that in wild populations of crabs shell perforations may lead to limited septicaemia potentially resulting in damage of internal tissues . Whether such natural infections lead to significant fatalities in crabs is still uncertain. Environ Technol, 2001 Nov, 22(11), 1287 - 93 Application of integrated chemical-physical processes modelling to aeration treatment of anaerobic digester liquors; Wentzel MC et al.; A three phase (aqueous/solid/gas) mixed weak acid/base kinetic model developed by Musvoto et al . is applied to simulate the physical and chemical processes that occur on aeration of anaerobic digester liquors . Included in the model are the kinetic reactions for (i) weak acid/base dissocations (water, carbonate, ammonium, phosphate, and short-chain fatty acids), (ii) precipitation of struvite, newberyite, amorphous calcium phosphate, calcium and magnesium carbonate, (iii) ion pair formation and (iv) stripping of CO2 and NH3 gases . To generate data for model application, batch aeration tests were conducted on two anaerobic digester liquors from (i) a spent wine upflow anaerobic sludge bed (UASB) digester and (a) a sewage sludge anaerobic digester . In the batch tests pH, Ca, Mg, PO4-P, free and saline ammonia (FSA)H2CO3* alkalinity (from which inorganic carbon is calculated) were measured . After establishing from the lIterature values for (i) weak acid/base equilbrium constants (pKa), (ii) weak add/base kinetic rate constants (K(ra)), and (iii) ion pair stability constants (pK(ST)), and trial and error determination of (iv) mineral solubility products (pK(SP)) (within the range reported in the literature), (v) ion pair kinetic rate constants (K(rIP)), (vi) mineral precipitation rate constants (Kppt) and (vii) gas stripping rates (K(rG)), a good correlation between predicted and measured data was obtained for all the parameters for both liquors . The solubility product values for the minerals that precipitated were the same for both liquors and fall in the range of values quoted in the literature, but the specific precipitation rate constants of the minerals differed for the two liquors. C R Acad Sci III, 2001 Dec, 324(12), 1067 - 76 Genomics and early cellular evolution . The origin of the DNA world; Forterre P; The sequencing of several genomes from each of the three domains of life (Archaea, Bacteria and Eukarya) has provided a huge amount of data that can be used to gain insight about early cellular evolution . Some features of the universal tree of life based on rRNA polygenies have been confirmed, such as the division of the cellular living world into three domains . The monophyly of each domain is supported by comparative genomics . However, the hyperthermophilic nature of the 'last universal common ancestor' (LUCA) is not confirmed . Comparative genomics has revealed that gene transfers have been (and still are) very frequent in genome evolution . Nevertheless, a core of informational genes appears more resistant to transfer, testifying for a close relationship between archaeal and eukaryal informational processes . This observation can be explained either by a common unique history between Archaea and Eukarya or by an atypical evolution of these systems in Bacteria . At the moment, comparative genomics still does not allow to choose between a simple LUCA, possibly with an RNA genome, or a complex LUCA, with a DNA genome and informational mechanisms similar to those of Archaea and Eukarya . Further comparative studies on informational mechanisms in the three domains should help to resolve this critical question . The role of viruses in the origin and evolution of DNA genomes also appears an area worth of active investigations . I suggest here that DNA and DNA replication mechanisms appeared first in the virus world before being transferred into cellular organisms. Biochem J, 2002 Feb 1, 361(Pt 3), 525 - 36 Evidence that the tandem-pleckstrin-homology-domain-containing protein TAPP1 interacts with Ptd(3,4)P2 and the multi-PDZ-domain-containing protein MUPP1 in vivo; Kimber WA et al.; PtdIns(3,4,5)P3 is an established second messenger of growth-factor and insulin-induced signalling pathways . There is increasing evidence that one of the immediate breakdown products of PtdIns(3,4,5)P3, namely PtdIns(3,4)P2, whose levels are elevated by numerous extracellular agonists, might also function as a signalling molecule . Recently, we identified two related pleckstrin-homology (PH)-domain-containing proteins, termed 'tandem-PH-domain-containing protein-1' (TAPP1) and TAPP2, which interacted in vitro with high affinity with PtdIns(3,4)P2, but did not bind PtdIns(3,4,5)P3 or other phosphoinositides . In the present study we demonstrate that stimulation of Swiss 3T3 or 293 cells with agonists that stimulate PtdIns(3,4)P2 production results in the marked translocation of TAPP1 to the plasma membrane . This recruitment is dependent on a functional PtdIns(3,4)P2-binding PH domain and is inhibited by wortmannin, a phosphoinositide 3-kinase inhibitor that prevents PtdIns(3,4)P2 generation . A search for proteins that interact with TAPP1 identified the multi-PDZ-containing protein termed 'MUPP1', a protein possessing 13 PDZ domains and no other known modular or catalytic domains {PDZ is postsynaptic density protein (PSD-95)/Drosophila disc large tumour suppressor (dlg)/tight junction protein (ZO1)} . We demonstrate that immunoprecipitation of endogenously expressed TAPP1 from 293-cell lysates results in the co-immunoprecipitation of endogenous MUPP1, indicating that these proteins are likely to interact with each other physiologically . We show that TAPP1 and TAPP2 interact with the 10th and 13th PDZ domain of MUPP1 through their C-terminal amino acids . The results of the present study suggest that TAPP1 and TAPP2 could function in cells as adapter proteins to recruit MUPP1, or other proteins that they may interact with, to the plasma membrane in response to signals that elevate PtdIns(3,4)P2. Pest Manag Sci, 2001 Dec, 57(12), 1135 - 41 Assessment of leaching potential of aldicarb and its metabolites using laboratory studies; Fava L et al.; The metabolites of pesticides can contaminate groundwater and pose a risk to human health when this water is used for drinking . This paper reports the results of a laboratory study on aldicarb and its main metabolites, aldicarb sulfone and aldicarb sulfoxide . Aldicarb and its metabolites showed Koc values (6-31) which were lower than that of atrazine (55), indicating that they are very mobile in soil . They are less persistent than atrazine (DT50 = 25 days), with DT50 values from less than 1 day and up to 12 days . Aldicarb behaved as a non-leacher, whereas its metabolites clearly showed the characteristics of leachers . Aged residue leaching experiments showed that aldicarb can occur at high concentrations in the leachate, together with its two metabolites . The leachate composition depends on the incubation time of the parent compound . Aldicarb and its metabolites can form various mixtures in groundwater on the basis of the time elapsing between the application of the insecticide and the first significant rainfall . This study confirms the characteristics of contaminants of aldicarb and especially its metabolites, as reported in the literature. Inorg Chem, 2002 Jan 28, 41(2), 309 - 16 Methanolysis of nitrocefin catalyzed by one and two Zn(2+) ions . A simplified model for class B beta-lactamases; Montoya-Pelaez PJ et al.; The methanolysis of nitrocefin (1) was investigated at 25 degrees C in anhydrous methanol as a function of {Zn(2+)} and as a model for the chemistry believed to occur in the active site of Zn(2+)(-)beta-lactamases . In the absence of metal ion, the vs rate constant profile shows a long plateau region having k(obs) = 9 x 10(-5) s(-1) between 7.5 and 12 generated after ionization of the COOH of 1 (kinetic pK(a) of 7.34) followed by attack of CH(3)O(-) with k(CH)3(O) = 1.18 M(-1) s(-1) . Strong catalysis of the methanolysis is observed at all pH values between 7.95 and 11.34 in the presence of as little as 0.05-3 mM Zn(2+) . Plots of the pseudo-first-order rate constant for methanolysis (k(obs)) as a function of reveal a saturation phenomenon indicative of formation of a 1:Zn(2+) complex, followed by a linear section indicative of the intervention of a second Zn(2+) ion in promoting the methanolysis of the complex . The two processes can be separated since the slope of the linear part of the plots gives the second-order rate constant (k(2)(obs)) for the second Zn process, while the intercept gives the spontaneous rate constant for methanolysis of 1:Zn(2+) (k(cat.)(obs)) . A plot of log k(cat.)(obs) vs is bell-shaped, maximizing at 10, with ascending and descending domains both first order in {CH(3)O(-)} . A plot of log k(2) vs gives an ascending domain second order in {CH(3)O(-)}, followed by a plateau above 9.5 . The data are analyzed in terms of a one-Zn and two-Zn model in which the former involves rate-limiting attack of methoxide on a 1:Zn(2+) complex up to the maximum and a spontaneous reaction of 1:Zn(2+):(CH(3)O(-))(2), while the latter involves bimolecular attack of Zn(2+)(CH(3)O(-))(2) on both 1:Zn(2+) and 1:Zn(2+):(CH(3)O(-))(2) . The relevance of these observations is discussed in terms of the currently accepted mechanism for hydrolyses of beta-lactams promoted by the Zn(2+)(-)beta-lactamases. Planta, 2001 Dec, 214(2), 304 - 13 Redox-regulation of the expression of the peroxide-detoxifying chloroplast 2-cys peroxiredoxin in the liverwort Riccia fluitans; Horling F et al.; 2-Cys peroxiredoxins (2-CPs) are H2O2- and alkyl hydroperoxide-detoxifying enzymes, and occur in animals, fungi, bacteria and higher plants . Here, the cDNA encoding a peroxiredoxin of a multicellular cryptogamic plant was first cloned from the liverwort Riccia fluitans L., and the dependence of its expression on the cellular redox state was analysed . The presence of an N-terminal targeting signal indicates that, like 2-CPs from higher plants, Riccia 2-CP is posttranslationally imported into chloroplasts . Addition of ascorbate and other reductants suppressed 2-CP gene expression and decre