Chemosphere, 2003 Apr, 51(2), 139 - 42 Non-degradable triazine substrates of atrazine and cyanuric acid hydrothermally and in supercritical water under the UV-illuminated photocatalytic cooperation; Horikoshi S et al.; Strong oxidation by titanium dioxide photocatalysis can occur by photodegradation of organic contaminants in air and water . Some endocrine disruptors such as 2,4-dichlorophenoxy acetic acid (;;; ), 2,4-dichlorophenol (;;; ), nonylphenol (; ), bisphenol A (), diethyl phthalate (; ), etc . which can be neither biodegraded by bacteria nor degraded thermally can be degraded by TiO(2) photocatalytic treatment . However, incomplete photomineralization partly occurred, when TiO(2) photocatalytic degradation is employed for the treatment of certain endocrine disruptors . For example, no atrazine pesticide having triazine skeleton can be completely mineralized even by a photocatalytic procedure; the photodegradation of atrazine ultimately stops at the intermediate step of cyanuric acid, which cannot be photodegraded even after long illumination times ().In this study, the decomposition of atrazine and cyanuric acid was carried out with a device combining photocatalytic degradation in supercritical water (scH(2)O) or hydrothermal water (hyH(2)O) . Atrazine and cyanuric acid can be degraded by the cooperation of either scH(2)O or hyH(2)O and UV illuminated TiO(2)-photocatalytic dispersed system under the fixed pressure of 23 MPa at 623 K or 683 K in a 120-ml Hastelloy batch reactor . The photocatalytic degradation method under high temperature and pressure has found appropriate for the photocatalytic oxidation of acetic acid and 2-chlorobiphenyl under continuous flow conditions at 160 degrees C and 20 atm () . In addition, the wet peroxide oxidation of PCBs by high temperature and pressure has been reported () . The main aims of this research are following . (i): the degradation of atrazine and cyanuric acid within the scH(2)O or hyH(2)O, (ii) the decomposition of atrazine and cyanuric acid catalyzed by TiO(2) particles under scH(2)O or hyH(2)O, and the synergistic effect for several reactions with TiO(2) and scH(2)O or hyH(2)O, and (iii) the mineralization yield of nitrogen and chlorine atoms concerning the chemical structures of atrazine or cyanuric acid (only nitrogen). Enferm Infecc Microbiol Clin, 2003 Feb, 21(2), 83 - 8 {Treatment approach for fungal infections in critically ill patients admitted to intensive care units: results of a multicenter survey}; Alvarez-Lerma F et al.; INTRODUCTION: Two consensus conferences taking place in the United States and Spain were organized to optimize diagnosis and treatment of Candida spp . infections . Among other results, clinical scenarios in which early prescription of antifungal agents is indicated were identified . OBJECTIVE: To determine the criteria followed by physicians for prescribing antifungal agents in critically ill patients in our country and to investigate adherence to the guidelines proposed by the consensus conferences . METHODS: A questionnaire was designed and directed to 4th- and 5th-year residents in intensive care medicine and to specialists in intensive care with training in infectious diseases or other medical areas . Four case reports for which expert consensus indicates early antifungal treatment were included in the questionnaire; 1) recurrent peritonitis secondary to perforation of the digestive tract, with mixed flora including fungi; 2) persistent febrile syndrome in a patient with multiple mucosal fungal colonizations treated with broad-spectrum antibiotics; 3) candiduria and pyuria in a febrile patient; and 4) candidemia . RESULTS: A total of 135 questionnaires from 45 different ICUs were returned (60% response rate) . In the candidemia and fungal peritonitis examples, early treatment with antifungal agents was indicated in 100% and 85.9% of responses, respectively, whereas for sepsis with multifocal candidiasis and candiduria associated with pyuria and fever, early treatment was prescribed in only 41.5% and 55.6% of responses, respectively . There were no significant differences in response with regard to degree of training of the physicians surveyed . Fluconazole prescription predominated, mainly at doses of 400 mg/day, in mixed peritonitis, disseminated candidiasis and candiduria, whereas amphotericin B lipid formulations were preferentially indicated in cases of candidemia . Antifungal treatment (early or late) was prescribed in all responses for candidemia, in 95.5% for mixed peritonitis (fungi and bacteria), in 79.5% for multifocal candidiasis in patients with persistent sepsis, and in 77.9% for candiduria with fever and pyuria . CONCLUSIONS: Adherence to recommendations from the consensus conferences was high among intensive medicine specialists, with no differences according to level of training in infectious diseases. Hist Cienc Saude Manguinhos, 2002 Sep-Dec, 9(3), 591 - 608 {Spontaneous generation and the hygienic concern about germ dissemination}; Caponi S; This paper analyzes the answer given by Pasteur to those who defended the spontaneous generation theory and evaluates the impact these arguments had on the history of public hygiene . Because of that, the article emphasizes transmission of germs, rather than the origin of life, which has been quite analyzed . It also discusses the criticism Pasteur made to naturalists' arguments and tries to show that it is possible to say there is a certain continuity between the air-related concerns on the part of classic hygienists, most of which infectionists, and Pasteur's concern with germs spread in the air. FEMS Microbiol Lett, 2003 Jan 21, 218(1), 15 - 21 Iron-regulated phenylalanyl-tRNA synthetase activity in Azotobacter vinelandii; Page WJ et al.; Azotobacter vinelandii strain UA22 was produced by pTn5luxAB mutagenesis, such that the promoterless luxAB genes were transcribed in an iron-repressible manner . Tn5luxAB was localized to a fragment of chromosomal DNA encoding the thrS, infC, rpmI, rplT, pheS and pheT genes, with Tn5 inserted in the 3'-end of pheS . The isolation of this mutation in an essential gene was possible because of polyploidy in Azotobacter, such that strain UA22 carried both wild-type and mutant alleles of pheS . Phenylalanyl-tRNA synthetase activity and PHES::luxAB reporter activity was partially repressed under iron-sufficient conditions and fully derepressed under iron-limited conditions . The ferric uptake regulator (Fur) bound to a DNA sequence immediately upstream of luxAB, within the pheS gene, but PHES::luxAB reporter activity was not affected by phenylalanine availability . This suggests there is novel regulation of pheST in A . vinelandii by iron availability. FEMS Microbiol Lett, 2003 Jan 21, 218(1), 9 - 14 Trans-translation and protein synthesis inhibitors; Vioque A et al.; Trans-translation is a process found in all bacteria, which contributes to the release of ribosomes that are stalled through a variety of causes, for example when the 3' end of a truncated mRNA lacking a stop codon is reached or at internal clusters of rare codons . Trans-translation requires tmRNA . Trans-translation is not essential for cell viability under laboratory conditions, but recently it has been shown that it can contribute to cell viability in the presence of protein synthesis inhibitors . In this minireview, we consider the connection between trans-translation and antibiotics and the potential of using trans-translation as a therapeutic target. Eur Arch Otorhinolaryngol, 2003 Feb, 260(2), 78 - 80 Epub 2002 Sep 13. Screening of middle ear effusion for the common sinus pathogen Bipolaris; Shin EJ et al.; Recently, a third of middle ear effusions have been shown to harbor fungal DNA by polymerase chain reaction (PCR) . This suggests that fungi, in addition to being an important sinus pathogen, may also play an important role in acute and serous otitis media . Bipolaris is an important sinus pathogen whose role in infections of the ear is unknown . In this study, we assessed if Bipolaris DNA was present in 19 middle ear effusions that were PCR positive for the presence of fungi . Primer pair specific for Bipolaris spicifera was tested against DNA from various bacterial and fungal species to demonstrate its specificity and was subsequently used on DNA isolated from effusions to determine if Bipolaris-specific DNA was present . None of the nineteen specimens tested positive for Bipolaris by PCR or standard culture technique . This suggests that while fungi may play an important role in otitis media, this study does not support a role for Bipolaris as a middle ear pathogen and may reflect regional differences in its prevalence. Sci STKE . 2003 Feb 11;2003(169):PL4. Single cell imaging of PI3K activity and glucose transporter insertion into the plasma membrane by dual color evanescent wave microscopy; Tengholm A et al.; Many signaling events involve the translocation of signaling molecules to or from the plasma membrane; however, suitable techniques to quantify the temporal relationships between such signaling events are lacking . Here, we describe an evanescent wave microscopy technique that allows parallel measurement of the recruitment and dissociation of cyan fluorescent protein (CFP) and yellow fluorescent protein (YFP) labeled proteins to and from the plasma membrane in individual living cells . The selective excitation of fluorescence in a zone less than 100 nm above a cover glass enables selective imaging within the plasma membrane of adherent cells, with markedly improved resolution, lower background, and minimal phototoxicity compared to confocal microscopy and other microscopy-based assays . In the microscope design we have developed, the beams from helium-cadmium (442 nm) and argon (514 nm) lasers are merged and focused through a dove prism at an angle that yields total internal reflection . In this configuration, evanescent wave-excited fluorescence at the glass-water interface can be detected with either high or low magnification, to allow for high-resolution imaging or the study of many cells in parallel . We applied this technique to make parallel measurements of the time-course of insulin-triggered activation of phosphatidylinositol 3-kinase (PI3K) and GLUT4 glucose transporter insertion into the plasma membrane of individual differentiated 3T3L1 adipocytes using a phosphatidylinositol-3,4,5-trisphosphate {PI(3,4,5)P(3)}-binding pleckstrin homology domain fused to CFP, and GLUT4 conjugated to YFP . The technique should have wide applicability to various cell types and diverse signaling processes. Exp Gerontol, 2003 Mar, 38(3), 245 - 52 A novel putative collaborator of p19ARF; Wadhwa R et al.; We performed a yeast interactive screen in search of p19(ARF)-binding partners and have isolated a novel serine rich protein that is assigned to human chromosome 4q35 and mouse chromosome 8 . The human and mouse proteins showed 84.2% homology . It is named CARF for its role as a putative Collaborator of ARF . CARF binds to both mouse and human ARF proteins . Its expression was detected in a variety of human tissues . The cDNA was expressed in bacteria and mammalian cells as a soluble and predominantly nucleoplasmic protein, respectively . CARF is a novel binding partner of ARF and might be involved in its p53-dependent or -independent tumor suppressive functions. Curr Opin Struct Biol, 2003 Feb, 13(1), 49 - 55 RNA-modifying enzymes; Ferre-D'Amare AR; A bewildering number of post-transcriptional modifications are introduced into cellular RNAs by enzymes that are often conserved among archaea, bacteria and eukaryotes . The modifications range from those with well-understood functions, such as tRNA aminoacylation, to widespread but more mysterious ones, such as pseudouridylation . Recent structure determinations have included two types of RNA nucleobase modifying enzyme: pseudouridine synthases and tRNA guanine transglycosylases. J Mol Biol, 2003 Feb 21, 326(3), 805 - 21 Control of directionality in L5 integrase-mediated site-specific recombination; Lewis JA et al.; Mycobacteriophage L5 is a temperate phage that forms lysogens in Mycobacterium smegmatis . These lysogens carry an integrated L5 prophage inserted at a specific chromosomal location and undergo subsequent excision during induction of lytic growth . Both the integrative and excisive site-specific recombination events are catalyzed by the phage-encoded tyrosine integrase (Int-L5) and require the host-encoded protein, mIHF . The directionality of these recombination events is determined by a second phage-encoded protein, Excise, the product of gene 36 (Xis-L5); integration occurs efficiently in the absence of Xis-L5 while excision is dependent upon it . We show here that Xis-L5 binds to attR DNA, introduces a DNA bend, and facilitates the formation of an intasome-R complex . This complex, which requires mIHF, Xis-L5 and Int-L5, readily recombines with a second intasome formed by Int-L5, mIHF and attL DNA (intasome-L) to generate the attP and attB products of excision . Xis-L5 also strongly inhibits Int-L5-mediated integrative recombination but does not prevent either the protein-DNA interactions that form the attP intasome (intasome-P) or the capture of attB, but acts later in the reaction presumably by preventing the formation of a recombinagenic synaptic intermediate . The mechanism of action of Xis-L5 appears to be purely architectural, influencing the assembly of protein-DNA structures solely through its DNA-binding and DNA-bending properties. Mol Microbiol, 2003 Feb, 47(4), 1075 - 89 Interaction of the sensor module of Mycobacterium tuberculosis H37Rv KdpD with members of the Lpr family; Steyn AJ et al.; The genetic and biochemical mechanisms by which Mycobacterium tuberculosis senses and responds to the complex environment that it encounters during infection and persistence within the host remain unknown . In a number of bacterial species, the Kdp signal transduction pathway appears to be the primary response to environmental osmotic stress, which is primarily mediated by K+ concentration in bacteria . We show that kdp encodes for components of a mycobacterial signalling pathway by demonstrating the K+ dependence of kdpFABC expression in both M . tuberculosis H37Rv and Mycobacterium smegmatis . To identify proteins of M . tuberculosis that participate in this signalling pathway, we used the N-terminal sensing module of the histidine kinase KdpD as bait in a yeast two-hybrid screen . We show that the sensing domain of KdpD interacts specifically with two membrane lipoproteins, LprJ (Rv1690) and LprF (Rv1368) . Overexpression of lprF and lprJ alleles in mycobacterial kdpF-lacZ reporter strains enabled us to identify alleles that modulate kdpFABC expression . By exploiting the yeast three-hybrid system, we have found that the histidine kinase domain of KdpD forms ternary complexes with LprF and LprJ and the sensing module of KdpD . Our results establish a role for membrane proteins in the Kdp signalling pathway and suggest that LprF and LprJ function as accessory or ligand-binding proteins that communicate directly with the sensing domain of KdpD to modulate kdp expression. Mol Plant Microbe Interact, 2003 Jan, 16(1), 14 - 24 Candidate defense genes from rice, barley, and maize and their association with qualitative and quantitative resistance in rice; Ramalingam J et al.; Candidate genes involved in both recognition (resistance gene analogs {RGAs}) and general plant defense (putative defense response {DR}) were used as molecular markers to test for association with resistance in rice to blast, bacterial blight (BB), sheath blight, and brown plant-hopper (BPH) . The 118 marker loci were either polymerase chain reaction-based RGA markers or restriction fragment length polymorphism (RFLP) markers that included RGAs or putative DR genes from rice, barley, and maize . The markers were placed on an existing RFLP map generated from a mapping population of 116 doubled haploid (DH) lines derived from a cross between an improved indica rice cultivar, IR64, and a traditional japonica cultivar, Azucena . Most of the RGAs and DR genes detected a single locus with variable copy number and mapped on different chromosomes . Clusters of RGAs were observed, most notably on chromosome 11 where many known blast and BB resistance genes and quantitative trait loci (QTL) for blast, BB, sheath blight, and BPH were located . Major resistance genes and QTL for blast and BB resistance located on different chromosomes were associated with several candidate genes . Six putative QTL for BB were located on chromosomes 2, 3, 5, 7, and 8 and nine QTL for BPH resistance were located to chromosomes 3, 4, 6, 11, and 12 . The alleles of QTL for BPH resistance were mostly from IR64 and each explained between 11.3 and 20.6% of the phenotypic variance . The alleles for BB resistance were only from the Azucena parent and each explained at least 8.4% of the variation . Several candidate RGA and DR gene markers were associated with QTL from the pathogens and pest . Several RGAs were mapped to BB QTL . Dihydrofolate reductase thymidylate synthase co-localized with two BPH QTL associated with plant response to feeding and also to blast QTL . Blast QTL also were associated with aldose reductase, oxalate oxidase, JAMyb (a jasmonic acid-induced Myb transcription factor), and peroxidase markers . The frame map provides reference points to select candidate genes for cosegregation analysis using other mapping populations, isogenic lines, and mutants. Adv Space Res, 2003, 31(1), 211 - 4 Pigment composition and concentrations within the plant (Ceratophyllum demersum L.) component of the STS-89 C.E.B.A.S . Mini-Module spaceflight experiment; Voeste D et al.; The Closed Equilibrated Biological Aquatic System (C.E.B.A.S.) Mini-Module, a Space Shuttle middeck locker payload which supports a variety of aquatic inhabitants (fish, snails, plants and bacteria) in an enclosed 8.6 L chamber, was tested for its biological stability in microgravity . The aquatic plant, Ceratophyllum demersum L., was critical for the vitality and functioning of this artificial mini-ecosystem . Its photosynthetic pigment concentrations were of interest due to their light harvesting and protective functions . "Post-flight" chlorophyll and carotenoid concentrations within Ceratophyllum apical segments were directly related to the quantities of light received in the experiments, with microgravity exposure (STS-89) failing to account for any significant deviation from ground control studies . Published by Elsevier Science Ltd on behalf of COSPAR. Eur J Med Res, 2003 Jan 28, 8(1), 33 - 46 Effect of periodontal therapy on sulcular sulphide level a longitudinal study; Gleissner C et al.; PURPOSE: The identification of active disease sites is a leading goal in basic periodontal research . Of toxic bacterial metabolites detectable in gingival crevicular fluid, volatile sulphur compounds (VSC) have been implicated in periodontal tissue destruction . Several bacteria associated with active destructive disease are capable of producing VSC, this fact supporting the idea of sulcular VSC being a possible marker of disease activity . A new portable sulphide monitor providing chairside information on sulcular sulphide level (SU) has been developed . The aim of this study was 1) to monitor the effect of mechanical therapy on SU and clinical parameters and 2) to clarify whether SU-measurements might have the potential to detect disease activity . METHODS: 34 patients (22 M, 12 F) with generalized or localized chronic periodontitis received periodontal treatment in a private practice consisting of an oral hygiene phase (HP) lasting several weeks, scaling and root planing (SRP), and flap surgery at sites >5 mm or with furcation involvement . Subjects were examined three times (1 week after the diagnosis was made, at the end of HP and at the 1st maintenance session 3 months after SRP) recording clinical parameters (clinical attachment loss CAL, probing depth PD, bleeding on probing intensity BI, plaque index PI) and sulcular sulphide level (SU) measured by the portable monitor as as digital score ranging from 0.0 (<10 superset 6 M of S superset 2-) to 5.0 (10 superset 2 M of S superset 2-) in increments of 0.5 . RESULTS: 23 patients (15 M, 8 F, 47.5 +/- 9 years) were included in the data analysis . Periodontal therapy resulted in a significant reduction of mean BI by 0.69 +/- 0.45, of mean PD by 1.39 +/- 0.33 mm and in a mean gain of attachment of 1.07 +/- 0.38 mm (p = 0.0001) . The clinical improvement was accompanied by a reduction of mean SU by 0.20 +/- 0.13 and of the mean percentage of SU-positive sites per patient (SUp) by 20.09 +/- 13 . SU-positive sites were located at all types of teeth . 67.9 % of SU-positive sites and 83.8 % of sites with a SU > 1 were found at the molars . 16.1 % of initially 579 SU-positive sites remained SU-positive . For these sites, BI reduction by treatment was significantly lower compared with SU-negative sites (p < 0.01) . SU was significantly correlated with PD, CAL, BI and PI (p = 0.0001) . Sensitivity and specificity of SU related to BOP (BI > 0) was 29.3 % and 96.8 %, respectively . CONCLUSIONS: Sulcular VSC as measured by the portable sulphide monitor may be an indicator of therapeutical success and periodontal stability . The sensor system offers chairside information on sulcular sulphide production at specific sites and is a valuable supplementation of traditional clinical methods in periodontal examination . More controlled longitudinal studies are needed to clarify the role of sulcular VSC at sites undergoing periodontal destruction. Nahrung, 2002 Dec, 46(6), 399 - 403 Dynamic rheological properties of dough as affected by amylases from various sources; Dogan IS; The effect of alpha-amylases from cereal, fungal, and bacterial sources on dough dynamic rheological properties was investigated . Dynamic rheological study of flour-and-water doughs during resting period showed significant changes in dough rheological properties as a function of alpha-amylases . Addition of alpha-amylases caused a time-dependent decrease in G', storage modulus . The enzyme action on starch during baking increased viscous flow properties . These changes were temperature-dependent . The thermal inactivation temperature of alpha-amylase plays an important role in modification of starch . Rheological changes in dough will alter the machinability of the dough and the quality of end products. Zhongguo Zhong Xi Yi Jie He Za Zhi, 2001 May, 21(5), 349 - 50 {Observation on effect of supplementary treatment by Astragalus injection in treating senile pulmonary tuberculosis patients}; Niu HR et al.; OBJECTIVE: To observe the clinical therapeutic supplementary effect of Astragalus injection (ASI) as anti-tuberculosis agents in treating senile tuberculosis (ST) . METHODS: Seventy-six ST patients were divided according to their hospitalization order into two groups randomly, 39 in the ASI group and 37 in the control group . The anti-tuberculosis regimen applied on all patients were HRE (S)Z for first treated patients and KHZ1321 TH for retreated patients . In the ASI group, ASI was given additionally by adding ASI 20 ml into 500 ml 5% glucose solution for intravenous dripping, once a day . The therapeutic course was 2 months . The changes of focal size, bacteria in sputum, and erythrocyte immune function (EIF) were observed before and after treatment, and the EIF obtained from 30 healthy subjects was taken for control . RESULTS: EIF in patients of both groups was lower than that in healthy subjects (P < 0.01) . Rosette rate of RBC-C3b receptor in both groups was all increased after treatment, the increment was higher in the ASI group than that in the control group significantly (P < 0.01) . After 2 months ASI treatment, the effective rate of focal absorption examined by X-ray was 84.6% and the negative conversion rate of bacteria in sputum was 79.4% . CONCLUSION: ASI has the effect of elevating erythrocyte immunity in senile pulmonary tuberculosis patients, it is able to enhance the therapeutic effect of treatment. Zhongguo Zhong Xi Yi Jie He Za Zhi, 2001 Feb, 21(2), 85 - 9 {Study on diagnosis and treatment of infections multiple organ dysfunction syndrome by integrated traditional Chinese and Western medicine}; Zhang SW et al.; OBJECTIVE: To explore the diagnostic and therapeutic approach of integrated traditional Chinese and western medicine (TCM-WM) on infectious multiple organ dysfunction syndrome/multiple system and organ failure (MODS/MSOF) for elevating the successful rate of rescuing the patients . METHODS: Diagnosis with western medicine and Syndrome Differentiation of TCM in 225 in-patients of acute infectious disease complicated with MODS/MSOF were conducted, and TCM treatment, based on western medical comprehensive treatment, was given to observe the effect and explore the mechanism of the TCM-WM therapy . RESULTS: Up to the end of 1998, 161 cases of the 225 cases were successfully cured and 64 died, the mortality being 28.4% . Among them, 58 out of 140 cases of MSOF died, the mortality was accounted for 41.4% . In 106 cases conformed to the diagnostic criteria of MSOF proposed by Professor Knaus WA, USA, 52 cases were cured successfully and 54 died, the mortality being 50.9% . CONCLUSION: TCM-WM treatment could elevate the therapeutic effect in treating MODS, the mechanism might be through improving the hemodynamic and hemorrheologic condition of patients to relieve nail-fold microcirculation disorder; influencing the levels of cytokine and inflammatory mediator, so as to alleviate the systemic inflammatory reaction, it might also abate the inhibited condition of gastro-intestinal motility, alleviate the intestinal flora imbalance, prevent intestinal bacteria and endotoxin malposition, and protect cells from peroxidation. Neurol India, 2002 Dec, 50(4), 470 - 2 Naegleria meningitis: a rare survival; Jain R et al.; Acute amebic meningoencephalitis caused by free-living amebae naegleria fowleri is extremely rare and uniformly fatal with only seven survivals reported till date . An interesting case of naegleria meningitis diagnosed by wet mount cytology of cerebrospinal fluid (CSF) and treated with amphoterecin B, rifampicin and ornidazole with complete recovery is presented . In cases of suspected pyogenic meningitis, if CSF staining, antigen detection or culture is negative for bacteria, a wet mount cytology of CSF for naegleria is suggested . Early treatment with amphoterecin B and rifampicin may improve survival. J Biol Chem, 2003 Apr 25, 278(17), 14647 - 56 Epub 2003 Feb 06. Impairment of the DNA binding activity of the TATA-binding protein renders the transcriptional function of Rvb2p/Tih2p, the yeast RuvB-like protein, essential for cell growth; Ohdate H et al.; In Saccharomyces cerevisiae, two highly conserved proteins, Rvb1p/Tih1p and Rvb2p/Tih2p, have been demonstrated to be major components of the chromatin-remodeling INO80 complex . The mammalian orthologues of these two proteins have been shown to physically associate with the TATA-binding protein (TBP) in vitro but not clearly in vivo . Here we show that yeast proteins interact with TBP under both conditions . To assess the functional importance of these interactions, we examined the effect of mutating both TIH2/RVB2 and SPT15, which encodes TBP, on yeast cell growth . Intriguingly, only those spt15 mutations that affected the ability of TBP to bind to the TATA box caused synthetic growth defects in a tih2-ts160 background . This suggests that Tih2p might be important in recruiting TBP to the promoter . A DNA microarray technique was used to identify genes differentially expressed in the tih2-ts160 strain grown at the restrictive temperature . Only 34 genes were significantly and reproducibly affected; some up-regulated and others down-regulated . We compared the transcription of several of these Tih2p target genes in both wild type and various mutant backgrounds . We found that the transcription of some genes depends on functions possessed by both Tih2p and TBP and that these functions are substantially impaired in the spt15/tih2-ts160 double mutants that confer synthetic growth defects. Bioresour Technol, 2003 Jun, 88(2), 131 - 6 Performance of a glucose fed periodic anaerobic baffled reactor under increasing organic loading conditions: 1 . Experimental results; Stamatelatou K et al.; The influence of the organic loading rate on the performance of an innovative reactor, the periodic anaerobic baffled reactor (PABR) was examined . A laboratory-scale PABR of four compartments being fed with a glucose based synthetic medium performed with high stability while the feed organic load was doubled from 12.5 to 25 and then to 50 gCOD/l . Finally the feed concentration was increased to 75 gCOD/l . The successive step changes in the feed concentration lasted for 20, 15, and 7 d, respectively . The COD removal efficiency of the PABR was satisfactory in the first two transitions (approximately 97.5 and 96%) . In the third transition (OLR=18.75 gCOD/l/d) the reactor failed as the pH dropped to 4 . The concentrations of butyric and valeric acids increased as the organic loading was increased and eventually they became greater than the concentration of acetic and propionic acids. Trends Biochem Sci, 2003 Feb, 28(2), 75 - 80 The insertion of palindromic repeats in the evolution of proteins; Claverie JM et al.; The current theory of protein evolution is that all contemporary proteins are derived from an ancestral subset . However, each new sequenced genome exhibits many genes with no detectable homologues in other species, leading to the paradoxical picture of a universal ancestor with more genes than any of its progeny . Standard explanations indicate that fast evolving genes might disappear into the 'twilight zone' of sequence similarity . Regardless of the size of the original ancestral subset, its origin and the potential mechanisms of its subsequent enlargement are rarely addressed . Sequencing of Rickettsia conorii genome recently led to the discovery of three families of repeat-mobile elements frequently inserted into the middle of protein coding genes . Although not yet identified in other species of bacteria, this discovery has provided the first clear evidence for the de novo creation of long protein segments (up to 50 amino acid residues) by repeat insertion . Based on previous results and theories on the coding potential of palindromic elements, we speculate that their insertion and mobility might have played a significant role in the early stages of protein evolution. Angew Chem Int Ed Engl, 2003 Feb 10, 42(6), 614 - 41 Biomineralization of unicellular organisms: an unusual membrane biochemistry for the production of inorganic nano- and microstructures; Bauerlein E; With evolution, Nature has ingeniously succeeded in giving rise to an impressive variety of inorganic structures . Every organism that synthesizes biogenic minerals does so in a form that is unique to that species . This biomineralization is apparently biologically controlled . It is thus expected that both the synthesis and the form of every specific biogenic mineral is genetically determined and controlled . An investigation of the mechanism of biomineralization has only become possible with the development of modern methods in molecular biology . Unicellular organisms such as magnetic bacteria, calcareous algae, and diatoms, all of which are amongst the simplest forms of life, are particularly suited to be investigated by these methods . Crystals and composites of proteins and amorphous inorganic polymers are formed as complex structures within these organisms; these structures are not known in conventional inorganic chemistry. J Mol Evol, 2003 Feb, 56(2), 151 - 61 Cost-minimization of amino acid usage; Seligmann H; The negative correlation between the frequencies of usage of amino acids and their biosynthetic cost suggests that organisms minimize costs of protein biosynthesis . Empirical results support that: (1) free-living organisms (Archaea, Bacteria, and Eucaryota) minimize the usage of heavy amino acids more than intracellular organisms (viruses, chloroplasts, and mitochondria), a result confirmed by comparing intracellular Bacteria with other Bacteria; (2) avoidance of amino acids with low impact on protein structure (Chou-Fasman indices) is greater than for those with equal molecular weight but greater structural impact: constraints on protein function limit cost-minimization; (3) amino acid weight minimization (WM) for a protein correlates positively with the protein's expression level and with its size; (4) preliminary results suggest that for different proteins, the evolutionary rate of amino acid replacements correlates negatively with WM in these proteins; (5) results suggest that WM decreases with genome-size; and (6) developmental rates correlate positively with WM (within primates and rodents), even after confounding factors were accounted for . Effects of biosynthetic cost-minimization at whole-organism levels vary with metabolic and ecological strategies . Biosynthetic cost-minimization is an adaptive hypothesis that yields a semi-mechanistic explanation for small differences in allele fitness. J Neurosci, 2003 Feb 1, 23(3), 955 - 60 Formation of complement membrane attack complex in mammalian cerebral cortex evokes seizures and neurodegeneration; Xiong ZQ et al.; The complement system consists of >30 proteins that interact in a carefully regulated manner to destroy invading bacteria and prevent the deposition of immune complexes in normal tissue . This complex system can be activated by diverse mechanisms proceeding through distinct pathways, yet all converge on a final common pathway in which five proteins assemble into a multimolecular complex, the membrane attack complex (MAC) . The MAC inserts into cell membranes to form a functional pore, resulting in ion flux and ultimately osmotic lysis . Immunohistochemical evidence of the MAC decorating neurons in cortical gray matter has been identified in multiple CNS diseases, yet the deleterious consequences, if any, of MAC deposition in the cortex of mammalian brain in vivo are unknown . Here we demonstrate that the sequential infusion of individual proteins of the membrane attack pathway (C5b6, C7, C8, and C9) into the hippocampus of awake, freely moving rats induced both behavioral and electrographic seizures as well as cytotoxicity . The onset of seizures occurred during or shortly after the infusion of C8/C9 . Neither seizures nor cytotoxicity resulted from the simultaneous infusion of all five proteins premixed in vitro . The requirement for the sequential infusion of all five proteins together with the temporal relationship of seizure onset to infusions of C8/C9 implies that the MAC was formed in vivo and triggered both seizures and cytotoxicity . Deposition of the complement MAC in cortical gray matter may contribute to epileptic seizures and cell death in diverse diseases of the human brain. J Immunol, 2003 Feb 15, 170(4), 2046 - 52 Persistence and turnover of antigen-specific CD4 T cells during chronic tuberculosis infection in the mouse; Winslow GM et al.; CD4 T cells are critical for resistance to Mycobacterium tuberculosis infection, but how effective T cell responses are maintained during chronic infection is not well understood . To address this question we examined the CD4 T cell response to a peptide from ESAT-6 during tuberculosis infection in the mouse . The ESAT-6(1-20)/IA(b)-specific CD4 T cell response in the lungs, mediastinal lymph nodes, and spleen reached maxima 3-4 wk postinfection, when the bacteria came under the control of the immune response . Once chronic infection was established, the relative frequencies of Ag-specific CD4 T cells were maintained at nearly constant levels for at least 160 days . ESAT-6(1-20)/IA(b)-specific CD4 T cells that responded in vitro expressed activation markers characteristic of chronically activated effector cells and used a limited Vbeta repertoire that was clonally stable in vivo for at least 12 wk . 5-Bromo-2-deoxyuridine incorporation studies indicated a relatively high rate of cell division among both total CD4 and ESAT-6(1-20)/IA(b)-specific CD4 T cells during acute infection, but the degree of 5-bromo-2-deoxyuridine incorporation by both the CD4 T cells and the Ag-specific cells declined at least 3-fold during chronic infection . The data indicate that the peripheral ESAT-6(1-20)/IA(b)-specific CD4 T cell response to M . tuberculosis is characterized during the acute phase of infection by a period of extensive proliferation, but once bacterial control is achieved, this is followed during chronic infection by an extended containment phase that is associated with a persistent response of activated, yet more slowly proliferating, T cells. J Clin Microbiol, 2003 Feb, 41(2), 863 - 6 Development of a 5' fluorogenic nuclease-based real-time PCR assay for quantitative detection of Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis; Yoshida A et al.; A 5' nuclease TaqMan PCR was developed for the quantitative detection of the periodontopathic bacteria Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis . The relative numbers of bacteria were measured by the comparative threshold cycle method . This simplified method is a way of obtaining the relative quantities of these organisms from specimens and of monitoring the effect of therapy. J Clin Microbiol, 2003 Feb, 41(2), 689 - 93 Detection of Francisella tularensis within infected mouse tissues by using a hand-held PCR thermocycler; Emanuel PA et al.; The diagnosis of human cases of tularemia often relies upon the demonstration of an antibody response to Francisella tularensis or the direct culturing of the bacteria from the patient . Antibody response is not detectable until 2 weeks or more after infection, and culturing requires special media and suspicion of tularemia . In addition, handling live Francisella poses a risk to laboratory personnel due to the highly infectious nature of this pathogen . In an effort to develop a rapid diagnostic assay for tularemia, we investigated the use of TaqMan 5' hydrolysis fluorogenic PCR to detect the organism in tissues of infected mice . Mice were infected to produce respiratory tularemia . The fopA and tul4 genes of F . tularensis were amplified from infected spleen, lung, liver, and kidney tissues sampled over a 5-day period . The samples were analyzed using the laboratory-based Applied Biosystems International 7900 and the Smiths Detection-Edgewood BioSeeq, a hand-held portable fluorescence thermocycler designed for use in the field . A comparison of culturing and PCR for detection of bacteria in infected tissues shows that culturing was more sensitive than PCR . However, the results for culture take 72 h, whereas PCR results were available within 4 h . PCR was able to detect infection in all the tissues tested . Lung tissue showed the earliest response at 2 days when tested with the ABI 7900 and in 3 days when tested with the BioSeeq . The results were in agreement between the ABI 7900 and the BioSeeq when presented with the same sample . Template preparation may account for the loss of sensitivity compared to culturing techniques . The hand-held BioSeeq thermocycler shows promise as an expedient means of forward diagnosis of infection in the field. J Biol Chem, 2003 Apr 18, 278(16), 14498 - 506 Epub 2003 Feb 06. Trafficking patterns of beta-arrestin and G protein-coupled receptors determined by the kinetics of beta-arrestin deubiquitination; Shenoy SK et al.; Agonist-dependent internalization of G protein-coupled receptors via clathrin-coated pits is dependent on the adaptor protein beta-arrestin, which interacts with elements of the endocytic machinery such as AP2 and clathrin . For the beta(2)-adrenergic receptor (beta(2)AR) this requires ubiquitination of beta-arrestin by E3 ubiquitin ligase, Mdm2 . Based on trafficking patterns and affinity of beta-arrestin, G protein-coupled receptors are categorized into two classes . For class A receptors (e.g . beta(2)AR), which recycle rapidly, beta-arrestin directs the receptors to clathrin-coated pits but does not internalize with them . For class B receptors (e.g . V2 vasopressin receptors), which recycle slowly, beta-arrestin internalizes with the receptor into endosomes . In COS-7 and human embryonic kidney (HEK)-293 cells, stimulation of the beta(2)AR or V2 vasopressin receptor leads, respectively, to transient or stable beta-arrestin ubiquitination . The time course of ubiquitination and deubiquitination of beta-arrestin correlates with its association with and dissociation from each type of receptor . Chimeric receptors, constructed by switching the cytoplasmic tails of the two classes of receptors (beta(2)AR and V2 vasopressin receptors), demonstrate reversal of the patterns of both beta-arrestin trafficking and beta-arrestin ubiquitination . To explore the functional consequences of beta-arrestin ubiquitination we constructed a yellow fluorescent protein-tagged beta-arrestin2-ubiquitin chimera that cannot be deubiquitinated by cellular deubiquitinases . This "permanently ubiquitinated" beta-arrestin did not dissociate from the beta(2)AR but rather internalized with it into endosomes, thus transforming this class A receptor into a class B receptor with respect to its trafficking pattern . Overexpression of this beta-arrestin ubiquitin chimera in HEK-293 cells also results in enhancement of beta(2)AR internalization and degradation . In the presence of N-ethylmaleimide (an inhibitor of deubiquitinating enzymes), coimmunoprecipitation of the receptor and beta-arrestin was increased dramatically, suggesting that deubiquitination of beta-arrestin triggers its dissociation from the receptor . Thus the ubiquitination status of beta-arrestin determines the stability of the receptor-beta-arrestin complex as well as the trafficking pattern of beta-arrestin. EMBO J, 2003 Feb 17, 22(4), 945 - 53 Mechanism of Tet(O)-mediated tetracycline resistance; Connell SR et al.; Tet(O) is an elongation factor-like protein which confers resistance to the protein synthesis inhibitor tetracycline by promoting the release of the drug from its inhibitory site on the ribosome . Here we investigated the interaction of Tet(O) with the elongating ribosome and show, using dimethyl sulfate (DMS) probing and binding assays, that it interacts preferentially with the post-translocational ribosome . Furthermore, using an XTP-dependent mutant of Tet(O), we demonstrated that Tet(O) induces conformational rearrangements within the ribosome which can be detected by EF-Tu, and manifested as a stimulation in the GTPase activity of this elongation factor . As such, these conformational changes probably involve the ribosomal GTPase-associated center and, accordingly, Tet(O) alters the DMS modification pattern of the L11 region . Additionally, tetracycline binding is associated with an E(a) of 58 kJ/mol . These results suggest a model where both Tet(O) and tetracycline induce a conformational change in functionally opposite directions and the Tet(O)-induced conformation persists after it has left the ribosome; this prevents rebinding of the drug while allowing productive A-site occupation by a ternary complex in the presence of tetracycline. Chem Biol, 2003 Jan, 10(1), 25 - 34 An unusual phylogenetic variation in the metal ion binding sites of porphobilinogen synthase; Jaffe EK; Porphobilinogen synthase (PBGS), which catalyzes the first common step in tetrapyrrole biosynthesis, contains a unique phylogenetic variation in the use of metal ions . Using sequence, structure, and enzymological information, this work codifies the phylogenetic segregation of metal utilization in PBGS from archaea, bacteria, and eucarya . All PBGS contain an active site metal binding sequence, determined herein to be either DXCXCX(Y/F)X(3)G(H/Q)CG or DXALDX(Y/F)X(3)G(H/Q)DG . The former dictates a requirement for zinc . Most PBGS that do not require zinc require magnesium and/or potassium instead . Most PBGS are also found to contain the binding determinants for an allosteric magnesium that resides outside the active site . The phylogenetic distribution of PBGS metal ion utilization suggests that the primordial PBGS required zinc and supports a hypothesis that the loss of the zinc site was concurrent with the advent of oxygenic photosynthesis. Allergol Immunopathol (Madr), 2003 Jan-Feb, 31(1), 7 - 13 Use of OM-85 BV in children suffering from recurrent respiratory tract infections and subnormal IgG subclass levels; Del-Rio-Navarro BE et al.; BACKGROUND: Recurrent acute respiratory tract infections (RARTIs) in children are related to IgG subclass deficiencies . The aim of the trial was to evaluate the effect of OM-85 BV in the number of RARTIs as well as in the IgG subclass levels . METHODS: This was a randomized, double-blind, placebo-controlled clinical trial . Patients of ages three to six years, having three or more documented ARTIs during the last six months with subnormal IgG subclass levels were included . Patients took either one capsule of OM-85 BV (3.5 mg) or placebo orally every day for ten consecutive days per month during three consecutive months . Patients were followed three further months without drug intake . IgG subclass levels were determined before and after treatment . RESULTS: IgG4 levels diminished after the OM-85 BV treatment (-3 {-8.0, -1.0} median difference {95 % CI} p < 0.05 by Wilcoxon test) . No other significant changes in IgG subclasses were observed . After six months the patients in the OM-85 BV group (n = 20) experienced 2.8 1.4 (mean SD) ARTIs, while the patients in the placebo group (n = 20) suffered 5.2 1.5 ARTIs (-2.4 {3.3, -1.5} mean difference {95 % CI} p < 0.001 by Student's t test) . Three patients with OM-85 BV had gastrointestinal events related to drug administration, as well as three placebo patients . CONCLUSION: This study demonstrated the clinical benefit of OM-85 BV in patients suffering from RARTIs and subnormal levels of IgG subclasses . This trial opens new perspectives in the research of the mechanism of action of OM-85 BV. Proc R Soc Lond B Biol Sci, 2002 Dec 22, 269(1509), 2509 - 18 Space and the persistence of male-killing endosymbionts in insect populations; Groenenboom MA et al.; Male-killing bacteria are bacteria that are transmitted vertically through the females of their insect hosts . They can distort the sex ratio of their hosts by killing infected male offspring . In nature, male-killing endosymbionts (male killers) often have a 100% efficient vertical transmission, and multiple male-killing bacteria infecting a single population are observed . We use different model formalisms to study these observations . In mean-field models a male killer with perfect transmission drives the host population to extinction, and coexistence between multiple male killers within one population is impossible; however, in spatially explicit models, both phenomena are readily observed . We show how the spatial pattern formation underlies these results . In the case of high transmission efficiencies, waves with a high density of male killers alternate with waves of mainly wild-type hosts . The male killers cause local extinction, but this creates an opportunity for uninfected hosts to re-invade these areas . Spatial pattern formation also creates an opportunity for two male killers to coexist within one population: different strains create spatial regions that are qualitatively different; these areas then serve as different niches, making coexistence possible. Biometals, 2003 Mar, 16(1), 185 - 97 Yeast, a model organism for iron and copper metabolism studies; De Freitas J et al.; Virtually all organisms on earth depend on transition metals for survival . Iron and copper are particularly important because they participate in vital electron transfer reactions, and are thus cofactors of many metabolic enzymes . Their ability to transfer electrons also render them toxic when present in excess . Disturbances of iron and copper steady-state levels can have profound effects on cellular metabolism, growth and development . It is critical to maintain these metals in a narrow range between utility and toxicity . Organisms ranging from bacteria and plants to mammals have developed sophisticated mechanisms to control metal homeostasis . In this review, we will present an overview of the current understanding of iron and copper metabolism in yeast, and the utility of yeast as a model organism to investigate iron and copper metabolism in mammals and plants. Am J Dent, 2002 Aug, 15(4), 236 - 43 Hierarchy of pulp capping and repair activities responsible for dentin bridge formation; Murray PE et al.; PURPOSE: The importance of pulp capping variables which mediate pulp repair activities and dentin bridge formation following pulp exposure are not well understood . Consequently, the aim of this study was to investigate the effects of pulp capping variables on tertiary dentin bridge formation . MATERIALS AND METHODS: Calcium hydroxide {Ca(OH)2}, resin-modified glass-ionomer (RMGI) and resin-based composite (RBC) were used to restore pulp exposures prepared in 161 nonhuman primate teeth, according to ISO usage guidelines . Teeth were collected from between 7 and 720 days to observe the pulp repair activities leading to dentin bridge formation . Bacteria were detected using McKays stain . Pulp activity was categorized histomorphometrically using ISO standards, and analyzed statistically using ANOVA statistics . RESULTS: The pulp capping variables in order of their relationship to dentin bridge area, from the most important to the least important were: time elapsed since pulp capping (P = 0.0009), pulp capping materials (P = 0.0252), bacterial microleakage (P = 0.0358) and area of operative debris in the dentin bridge (0.0362) . A further 10 pulp capping variables were found to be less important . Relationships between pulp capping materials, operative debris, tunnel defects, bacterial microleakage, inflammation and dentin bridge formation were observed . Bacteria contaminated 18.6% of RBC, 22.2% of RMGI and 47.0% of Ca(OH)2 capped pulps. Wei Sheng Yan Jiu, 2002 Oct, 31(5), 386 - 8 {Application of an ozone disinfection apparatus for small drinking water supply systems in rural areas}; Sun X et al.; To design an ozone disinfection apparatus which is inexpensive and to practical for small drinking water supply system in rural areas . And test its effectiveness . Designed apparatus must be rapid, low cost, high automatic and easy for operation . The key technologies are to create ozone of high concentration and to mix it with water very well, and add it into main water pipe automatically . The results showed: In laboratory, at the condition of ozone of 0.2 mg/l for 3 minutes, all germs and bacteria coli were killed . In the field study, the water samples from three villages met the standard for drinking water very well . THE CONCLUSIONS: The apparatus reached the design requirement . An innovative part of the study is that we have improved a device for mixture of ozone with water . The laboratory and field study have proved the apparatus is low cost, high effectiveness, and easy to operate u . It is very useful for small drinking water supply system in rural areas. Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi, 2001, 19(4), 229 - 32 {Experimental study on the pathogenesis of Entamoeba gingivalis}; Liu GY et al.; OBJECTIVE: To study the pathogenesis of Entamoeba gingivalis (E . g.) and its relation to periodontal diseases . METHODS: Rats were treated with immuno-inhibitor for one week and the neck of incisor teeth of the rats was bound with steel wire . They were randomly divided into three groups: the first group was infected by E . g . in the periodontal tissue, the second group was infected by symbiotic bacteria (s . b.), and the third group was given physiological saline as control . Observation on the periodontal inflammation was made for each group of rats, and the purulent secretion from periodontal abscess was examined for living pathogens . RESULTS: The incidence of periodontal diseases in rats infected by E . g . was higher than that of symbiotic bacteria group and that of control (P < 0.05), the incidence of periodontal diseases in rats infected by s . b . was higher than that of control group (P < 0.05) . Living pathogens were found in the abscess liquid . CONCLUSION: E . g . is an opportunistic pathogen, which, together with synergistic symbiotic bacteria, can cause periodontal diseases in hosts with low immunity. Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi, 2001, 19(3), 157 - 9 {Cloning and identification of an unknown gene encoding 10.6 kDa protein of Schistosoma japonicum}; Shen JJ et al.; OBJECTIVE: To screen a new schistosome vaccine candidate . METHODS: Schistosoma japonicum adult cDNA library was screened using sera from immune rabbits vaccinated with irradiated cercariae and monoclonal antibodies against membrane antigen of S . japonicum schistosomula . Three different fragments of S . japonicum cDNA genes were cloned into pGEM-T vector . The sequences of the inserts were determined using an automatic DNA sequencer and were analysed using Blast program . One of the unknown genes (B8) was selected and its ORF sequence (291 bp) was subcloned into eukaryotic expression vector . The recombinant plasmids were identified by restrictive enzymes and PCR amplification . The positive recombinant plasmids (pBK/SjB8) were transformed into host bacteria XL1-blue, and were then induced by IPTG for expression . SDS-PAGE and Western blotting analysis of total cellular protein from the bacteria were performed to detect the gene products . RESULTS: The results demonstrated that ORF of SjB8 gene was subcloned into the plasmid pBK-CMV and could express as fusion protein in XL1-blue . The results of SDS-PAGE and Western-blot also showed that the molecular weight of the fusion protein with 3 kDa beta-galactosidase was approximately 13.6 kDa and the actual molecular weights of the SjB8 was 10.6 kDa . The expressed fusion product of pBK/Sj-B8 could be recognized by immune serum and McAb . CONCLUSION: A new gene of S . japonicum vaccine candidate (SjB8) was cloned into eukaryotic expression vector pBK-CMV and could express 10.6 kDa schistosome protein . The results provide foundation for further study of the protein for its possibility as candidate vaccine. Pathobiology, 2002-2003, 70(3), 164 - 9 Anti-interleukin-18 therapy in murine models of inflammatory bowel disease; Lochner M et al.; Interleukin (IL)-18 is a cytokine with a broad array of effector functions, the most prominent of which is to act synergistically with IL-12 in interferon-gamma production and the induction of a strong T-helper-1-mediated immune response . In addition, IL-18 also upregulates the production of proinflammatory cytokines such as IL-1 and tumor necrosis factor-alpha . Analysis of IL-18-deficient mice revealed an important role of IL-18 in the activation of macrophages and natural killer cells in the context of infection with intracellular bacteria or parasites . In humans, it was reported that IL-18 is elevated at sites of inflammation in inflammatory bowel disease (IBD), particularly in Crohn's disease, suggesting a possible role for IL-18 in the development and persistence of IBD . In this review we summarize recent findings on the functional role of IL-18 in the pathogenesis of colitis with a special focus on murine models of IBD . The neutralizing mouse anti-mouse IL-18 antibodies generated in our group should facilitate the evaluation of the efficiency of therapeutic blockade of endogenous IL-18 in chronic mouse models of colitis besides the use of recombinant forms of the inhibitory IL-18-binding protein . Appl Environ Microbiol, 2003 Feb, 69(2), 1043 - 50 Seasonal changes in fungal production and biomass on standing dead Scirpus lacustris litter in a northern prairie wetland; Verma B et al.; Decaying macrophytes are an important source of carbon and nutrients in fungal and bacterial communities of northern prairie wetlands . Dead macrophytes do not collapse into the water column immediately after death, and decomposition by fungi and bacteria begins while the plants are standing . The seasonal variations in fungal biomass and production on Scirpus lacustris stems, both above and below water, were measured to assess which environmental factors were dominant in affecting these variations in a typical prairie wetland . Fungal biomass and production were measured from early May to November, just prior to freeze-up . Fungal decomposition began and was greatest in the spring despite low water temperatures . The fungal production, as measured by the incorporation of {1-(14)C}acetate into ergosterol, ranged from 1.8 to 376 microg of C g of ash-free dry mass (AFDM)(-1) day(-1), and the biomass, as estimated by using ergosterol, ranged from nondetectable to 5.8 mg of C g of AFDM(-1) . There was no significant difference in biomass or production between aerial and submerged portions of Scirpus stems . The water temperature was correlated with fungal production (r = 0.7, P < 0.005) for aerial stem pieces but not for submerged pieces . However, in laboratory experiments water temperature had a measurable effect on both biomass and production in submerged stem pieces . Changes in fungal biomass and productivity on freshly cut green Scirpus stems decaying in the water either exposed to natural solar radiation or protected from UV radiation were monitored over the summer . There was no significant difference in either fungal biomass (P = 0.76) or production (P = 0.96) between the two light treatments . The fungal biomass and rates of production were within the lower range of the values reported elsewhere, probably as a result of the colder climate and perhaps the lower lability of Scirpus stems compared to the labilities of the leaves and different macrophytes examined in other studies performed at lower latitudes. Curr Top Med Chem, 2003, 3(3), 249 - 82 Fluoroquinolones: action and resistance; Drlica K et al.; Fluoroquinolones trap gyrase and topoisomerase IV on DNA as ternary complexes that block the movement of replication forks and transcription complexes . Studies with resistant mutants indicate that during complex formation quinolones bind to a surface alpha-helix of the GyrA and ParC proteins . Lethal action is a distinct event that is proposed to arise from release of DNA breaks from the ternary complexes . Many bacterial pathogens are exhibiting resistance due to alterations in drug permeability, drug efflux, gyrase-protecting proteins, and target topoisomerases . When selection of resistant mutants is described in terms of fluoroquinolone concentration, a threshold (mutant prevention concentration, MPC) can be defined for restricting the development of resistance . MPC varies among fluoroquinolones and pathogens; when combined with pharmacokinetics, MPC can be used to identify compounds least likely to enrich mutant subpopulations . Use of suboptimal doses and compounds erodes the efficacy of the class as a whole because resistance to one quinolone reduces susceptibility to others and/or increases the frequency at which resistance develops . When using fluoroquinolones in combination therapy, the development of resistance may be minimized by optimizing regimens for pharmacokinetic overlap. Curr Pharm Biotechnol, 2003 Feb, 4(1), 51 - 67 Recombinant secretory immunoglobulin A in passive immunotherapy: linking immunology and biotechnology; Corthesy B; The use of monoclonal antibodies has become routine in the research and diagnostic laboratories, but the potential of antibody molecules in public health and medical applications is still far from its maximum . Most infections begin at mucosal surfaces, and this is certainly not only a stroke of good fortune if mother's milk serves as a natural delivery vehicle for antibodies protecting the gastrointestinal tract of nursing infants . Mammary gland or other mucous secretions containing numerous antibody specificities provide an efficient mean to immediately protect a mucosal surface against pathogens, which have never been encountered by the host . From a public health perspective, topical passive immunization of mucosal surfaces with monoclonal antibodies can block entry and transmission of bacteria, viruses, fungi and parasites that infect humans, and thus defeat some key immune evasion strategies designed by many pathogens . The chief antibody on most mucosal surfaces is secretory immunoglobulin A (SIgA), a polypeptide complex comprising dimeric IgA, the connecting J chain, and the secretory component . The molecular stability, tetravalency, and strong anti-inflammatory properties make SIgA particularly well suited to fulfill the function of passive protective immunity when applied exogenously to mucosal surfaces . The review will give an overview of the basic concepts underlying mucosal immunity, present the molecular mechanisms whereby SIgA prevents mucosal infections, cover the last advances in the topic of recombinant SIgA production, and examine how structure-function relationship in SIgA will help designing molecules with novel properties for passive immunotherapy. Curr Cancer Drug Targets, 2003 Feb, 3(1), 57 - 65 The prodigiosins: a new family of anticancer drugs; Montaner B et al.; Apoptosis is involved in the action of several (and perhaps all) cancer-chemotherapeutic agents . Prodigiosins, a family of natural red pigments characterized by a common pyrrolylpyrromethene skeleton, are produced by various bacteria . Three members of the prodigiosin family, viz . prodigiosin (PG), undecylprodigiosin (UP) and cycloprodigiosin hydrochloride (cPrG.HCl), have immunosuppressive properties and apoptotic effects on cancer cells in vitro and in vivo . Their cytotoxic effect is attributed to the presence of the C-6 methoxy substituent . The A-pyrrole ring plays a key role in both the copper nuclease activity and the cytotoxicity of prodigiosins . Here, we have reviewed the pharmacological activity of PG and related compounds, including novel synthetic PG-derivatives with lower toxicity . The mechanism of action for these molecules is a current topic in biomedicine . The molecular targets of prodigiosins are also discussed. Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi, 1999 Jun 30, 13(2), 128 - 9 {Establishment of hybridomas secreting monoclonal antibodies to HIV-gp41 and HCV-NS3}; Zhang S et al.; OBJECTIVE: To establish the monoclonal antibodies of HIV and HCV for developing serological diagnostic method . METHODS: The antigens of HIV gp41 and HCV NS3 expressed from bacteria were mixed to immunize BALB/c mice . The hybridoma cell lines were obtained by fusing SP2/0 myeloma cells with spleen cells from immunized mice . RESULTS: Four hybridoma cell lines secreting specific antibodies against gp41 of HIV and six hybridoma cell lines secreting antibodies to NS3 of HCV were obtained . ELISA sandwich method for detection of viral antigens with McAbs was developed . CONCLUSION: A quick and convenient method for obtaining McAbs was developed using mixture of different antigens for immunizition of mice . The monoclonal antibodies had high senusitivity and were stably secreted by hybridoma cell lines . This technique is worthy to be popularized. Biotechnol Bioeng, 2003 Apr 5, 82(1), 54 - 61 A new VFA sensor technique for anaerobic reactor systems; Pind PF et al.; A key parameter for understanding and controlling the anaerobic biogas process is the concentration of volatile fatty acids (VFA) . However, this information has so far been limited to off-line measurements using labor-intensive methods . We have developed a new technique that has made it possible to monitor VFA on-line in one of the most difficult media: animal slurry or manure . A novel in situ filtration technique has made it possible to perform microfiltration inside a reactor system . This filter enables sampling from closed reactor systems without large-scale pumping and filters . Furthermore, due to its small size it can be placed in lab-scale reactors without disturbing the process . Using this filtration technique together with commercially available membrane filters we have constructed a VFA sensor system that can perform automatic analysis of animal slurry at a frequency as high as every 15 minutes . Reproducibility and recovery factors of the entire system have been determined . The VFA sensor has been tested for a period of more than 60 days with more than 1,000 samples on both a full-scale biogas plant and lab-scale reactors . The measuring range covers specific measurements of acetate, propionate, iso-/n-butyrate and iso-/n-valerate ranging from 0.1 to 50 mM (6-3,000 mg) . The measuring range could readily be expanded to more components and both lower and higher concentrations if desired . In addition to the new VFA sensor system, test results from development and testing of the in situ filtration technique are being presented is this article . Eur J Med Res, 2002 Nov 25, 7(11), 491 - 501 Evaluation of sulcular sulphide level monitoring using a portable sensor system; Gleissner C et al.; PURPOSE: Volatile sulphur compounds (VSC) such as hydrogen sulphide and methyl mercaptan are toxic bacterial metabolites considered to contribute to the progression of periodontitis . However, the quantitative determination of sulcular VSC concentration requires costly time-consuming laboratory methods limiting its evaluation in large clinical trials . Recently, a portable monitor with a sulphide microsensor has been developed to measure sulcular VSC levels (SU) . This cross-sectional study evaluated 1) the clinical usage of the sulphide sensor system, 2) the reproducibility of SU-measurements, 3) the distribution of SU-values in periodontal health and disease and 4) the relationship between SU and clinical parameters documenting periodontal disease severity . METHODS: The specificity, the measuring range and the reproducibility of the sensor system were tested with reference solutions in vitro . 10 probands with healthy gingiva, 23 patients with gingivitis and 30 patients with chronic periodontitis were examined recording periodontal parameters (clinical attachment loss, CAL, probing depth, PD, bleeding on probing intensity BI, plaque and gingival index PI, GI) and sulcular sulphide level (SU) measured by the portable monitor as a digital score ranging from 0.0 (< 10 superset -6 M of S2-) to 5.0 (10 superset -2 M of S superset 2-) in increments of 0.5 . RESULTS: The in vitro testing confirmed the sensor s specificity for sulphide ions . The intraexaminer agreement (Pearson correlation coefficient) was 0.91 between two measurements (p < 0.05) . However, positive SU-values decreased with measurement repetition due to the volatility of the sulphur compounds . SU and the percentage of positive SU-values per patient were correlated with disease severity . There were no positive SU-values in healthy probands, a mean SU of 0.11 +/- 0.08 in gingivitis and of 0.22 +/- 0.15 in periodontitis (p <0.001) . SU was significantly correlated with PD in gingivitis, PD and CAL in periodontitis and the bleeding index in the analysis of all data . CONCLUSION: The portable sulphide monitor is a reliable method for the chairside determination of sulcular sulphide level in periodontal disease and in spite of its limitations a valuable supplementation of traditional clinical examination methods . It offers the possibility of relating in vitro results on the toxicity of VSC to clinical findings and might provide new insight into the impact of sulcular sulphide on the pathophysiology of periodontitis. Ecotoxicol Environ Saf, 2002 Oct, 53(2), 226 - 37 Confounding factors in bioassays with freshwater and marine organisms; Postma JF et al.; The use of bioassays in ecological risk assessments often raises questions about the causative factors, and insight into the possibility that confounding factors, such as pH or increased ammonia concentrations, might be responsible for the observed toxicity is needed . It was decided to develop a practical approach for the Dutch situation, in which a first screening is carried out based on provisional criteria . In collecting the required data, dozens of experiments were performed, while the scientific literature was searched for additional information . It is concluded that the provisional criteria specified are at present useful tools in interpreting results of bioassays . Depending on the outcome and the aim of the research, it might be necessary to further reduce uncertainties in the interpretation . This might require some additional experiments, using alternative controls or test procedures or altering the composition of the original sample. Ecotoxicol Environ Saf, 2002 Oct, 53(2), 221 - 5 Bioluminescence assays: effects of quinones and phenols; Kudryasheva N et al.; The influence of a series of quinones and phenols on bacteria bioluminescence systems was investigated . Three bioluminescence systems used in ecological monitoring were compared: (1) water-soluble; (2) immobilized in starch gel coupled enzyme systems: NADH:FMN-oxidoreductase-luciferase; (3) luminescent bacteria . Bioluminescence inhibition constants of quinones and phenols and bioluminescence induction periods were compared . These kinetic parameters are proportional to quinone concentrations and depend on the quinone redox potential . Different effects of the substances are related to structure and properties of the bioluminescence systems . The set of bioluminescence assays for quinones and phenols monitoring should include two bioluminescence systems: 1 (or 2) and 3. Cell Mol Life Sci, 2002 Dec, 59(12), 2088 - 96 The functions of mucosal T cells in containing the indigenous commensal flora of the intestine; Macpherson AJ et al.; There is an immense load of non-pathogenic commensal bacteria in the distal small intestine and the colon of mammals . The physical barrier that prevents penetration (translocation) of these organisms into the body is a simple epithelium comprised of the single enterocyte/colonocyte cell layer with its overlying mucus . In this review, we discuss the roles of intestinal T cells in initiating and regulating innate and adaptive mucosal immune responses of the mucosal immune system that avoid or limit penetration of the commensal intestinal bacteria. J Air Waste Manag Assoc, 2003 Jan, 53(1), 21 - 31 Pig slurry concentration by vacuum evaporation: influence of previous mesophilic anaerobic digestion process; Bonmati A et al.; Water can be removed from pig slurry by evaporation, through the application of wasted heat from a power plant or from other processes . Apart from obtaining a concentrate with an obviously higher nutrient concentration than the original slurry, another objective of water removal is to obtain water as condensate, which could be reused . The objective of this work was to study the vacuum evaporation of pig slurry liquid fraction and to evaluate condensate composition as a function of both pH (4, 5, and 6) and pig slurry type (fresh slurry and anaerobically digested slurry) . Batch experiments showed that condensate characteristics, total ammonia nitrogen (NH3-N), volatile fatty acids (VFA), and chemical oxygen demand were strongly dependent on initial slurry pH . In addition to producing part of the required thermal energy, previous anaerobic digestion presented several other clear advantages . The consumption of VFA and other volatile organic compounds during anaerobic digestion reduced the volatilization of organic matter in the evaporation treatment and, consequently, provided a higher quality condensate. Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi, 2000, 18(2), 79 - 83 {Studies on the recurrent attacks of acute adenolymphangitis due to Malayan filariasis}; Shi ZJ et al.; OBJECTIVE: To explore the role of filarial and bacterial infections in the recurrent attacks of acute adenolymphangitis due to malayan fialriasis . METHODS: 1 . To observe the seasonal fluctuation of acute attacks by performing monthly follow-up on patients with history of acute attacks in recent years . 2 . To study the relationship between bacterial infection and filarial adenolymphangitis by performing bacteria culture and anti-streptolysin O test . 3 . To investigate the variation of acute attacks by controlling filariasis transmission or by treating patients with a history of recurrent acute attacks . RESULTS: 1 . The peak of acute attacks in patients coincided with the peak of vector transmission season . 2 . Of the 97 cases examined by bacteria culture, 90 cases were negative; of the 255 cases examined by anti-streptolysin O test, the titres in 94.1% (143/152) of the cases with first attack and simple adenolymphangitis were within normal limits, however, the titres in 27.2% (28/103) of the cases complicated with elephantiasis were increased . 3 . The acute attack rate of adenolymphangitis per year reduced significantly in cases with first attack and simple adenolymphangitis after effective control of filariasis transmission . 4 . There was no evidence of the reduction of acute attacks by treating patients with DEC alone . CONCLUSION: In malayan filariasis endemic areas, the main causes of recurrent attacks of acute adenolymphangitis might be the repeated filarial infections due to the persistence of filariasis transmission. Laryngoscope, 2003 Feb, 113(2), 322 - 7 Antigenic as well as nonantigenic stimuli induce similar middle ear responses in the rat; Tonnaer EL et al.; OBJECTIVES/HYPOTHESIS: The observation that during otitis media many different types of micro-organisms have been cultured from effusions indicate that, once present in the middle ear cavity, most types of micro-organisms are able to trigger an inflammatory reaction leading to otitis media . The present study was designed to determine the middle ear response after injection of different substances into the middle ear cavity . STUDY DESIGN: To determine whether and to what extent an inflammatory response of the middle ear depends on the entering agent, the response in the tympanic cavity was studied by otomicroscopy and histological examination after inoculation of various substances . METHODS: Lewis rats were inoculated in transtympanic fashion either with live or heat-killed bacteria (pathogenic and nonpathogenic), Keyhole limpet hemocyanin, active charcoal, or saline . The mucosal response of the challenged middle ears was studied histologically . RESULTS: Irrespective of the inoculated substance, no essential differences in the mucosal response were found . The intensity of the inflammatory response was greater when live bacteria were inoculated . CONCLUSIONS: The present study demonstrates that any substance reaching the middle ear cavity is likely to induce otitis media . These observations emphasize the role of the eustachian tube as "porte d'entree" in the pathogenesis of this disorder . Determination of specific aspects of the eustachian tube involved in protection or in facilitating bacterial translocation will be important for the understanding of the pathogenesis of otitis media and the subsequent development of new therapeutic strategies . In addition, elucidation of bacterial factors involved in the process of colonization and translocation will be of equal importance. Neuroepidemiology, 2003 Jan-Feb, 22(1), 95 - 9 Ambient air quality and occurrence of multiple sclerosis relapse; Oikonen M et al.; Infectious viruses and bacteria can trigger multiple sclerosis (MS) exacerbations . Seasonally changing concentrations of ambient air pollutants are known to predispose to transmissible infections, to induce systemic immune responses and to enhance existing peripheral inflammation . Ambient air quality and monthly MS relapse occurrence in south-western Finland were compared by multivariate logistic regression . The odds ratio of the risk of a relapse onset was over fourfold (4.143, p < 0.001) when the concentration of inhalable particulate matter (PM(10)) was at the highest quartile . Inhalable airborne particulate matter concentrations were connected to relapse occurrence . Poor air quality may enhance the seasonal changes in MS relapse occurrence by an increased susceptibility to transmissible infections . Biochem Biophys Res Commun, 2003 Feb 7, 301(2), 392 - 8 Nedd8-modification of Cul1 is promoted by Roc1 as a Nedd8-E3 ligase and regulates its stability; Morimoto M et al.; SCF is a ubiquitin ligase and is composed of Skp1, Cul1, F-box protein, and Roc1 . The catalytic site of the SCF is the Cul1/Roc1 complex and RING-finger protein Roc1 . It was shown earlier that when Cul1 was co-expressed with Roc1 in Sf-9 cells in a baculovirus protein expression system, Cul1 was highly neddylated in the cell, suggesting that Roc1 may function as a Nedd8-E3 ligase . However, there is no direct evidence that Roc1 is a Nedd8-E3 in an in vitro enzyme system . Here we have shown that Roc1 binds to Ubc12, E2 for Nedd8, but not to Ubc9, E2 for SUMO-1 and Roc1 RING-finger mutant, H77A, did not bind to Ubc12 . In in vitro neddylation system using purified Cul1/Roc1 complex expressed in bacteria, Roc1 promotes neddylation of Cul1 . These results demonstrate that Roc1 functions as a Nedd8-E3 ligase toward Cul1 . Furthermore, Roc1 and Cul1 were ubiquitinylated in a manner dependent on the neddylation of Cul1 in vitro . In addition, Cul1 was degraded through the ubiquitin-proteasome pathway, and a non-neddylated mutant Cul1, K720R, was more stable than wild-type in intact cells . Thus, neddylation of Cul1 might regulate SCF function negatively via degradation of Cul1/Roc1 complex. Biochem Biophys Res Commun, 2003 Feb 14, 301(3), 693 - 8 Critical role of N-terminal N-glycosylation for proper folding of the human formyl peptide receptor; Wenzel-Seifert K et al.; The human formyl peptide receptor (FPR) is N-glycosylated and activates phagocytes via G(i)-proteins . The FPR expressed with G(i)alpha(2)beta(1)gamma(2) in Sf9 insect cells exhibits high constitutive activity as assessed by strong inhibitory effects of an inverse agonist and Na(+) on basal guanosine 5(')-O-(3-thiotriphosphate) (GTPgammaS) binding . The aim of our study was to analyze the role of N-glycosylation in FPR function . Site-directed mutagenesis of extracellular Asn residues prevented FPR glycosylation but not FPR expression in Sf9 membranes . However, in terms of high-affinity agonist binding, kinetics of GTPgammaS binding, number of G(i)-proteins activated, and constitutive activity, non-glycosylated FPR was much less active than native FPR . FPR-Asn4Gln/Asn10Gln/Asn179Gln and FPR-Asn4Gln/Asn10/Gln exhibited similar defects . Our data indicate that N-glycosylation of N-terminal Asn4 and Asn10 but not of Asn179 in the second extracellular loop is essential for proper folding and, hence, function of FPR . FPR deglycosylation by bacterial glycosidases could be a mechanism by which bacteria compromise host defense. Trends Microbiol, 2002 Dec, 10(12), 580 - 4 Environmental regulation of mutation rates at specific sites; Massey RC et al.; Recent studies on bacterial adaptation to stress suggest that bacteria can regulate the generation of mutations at specific sites in response to environmental conditions . Here, we review these findings and discuss the circumstances under which these mechanisms might prove advantageous. Farmaco, 2002 Dec, 57(12), 1015 - 8 Synthesis and antifungal properties of N-{(1,1'-biphenyl)-4-ylmethyl}-1H-imidazol-1-amine derivatives; Setzu MG et al.; In the course of a study on 1H-imidazol-1-amine derivatives as antifungal agents, we found that N-{(1,1'-biphenyl)-4-ylmethyl}-N-{(2,4-dichlorophenyl)methyl}-1H-imidazol-1-amine (1a) exhibited promising activities . In order to explore more in detail the structure-activity relationship of this new class of antifungal agents, we report now the synthesis and the biological activity of new analogues (1b-k) of compound 1a . The synthesis was performed using N-{(1,1'-biphenyl)-4-ylmethyl}-1H-imidazol-1-amine as starting material which was reacted with the proper arylmethyl halide . Most of the newly synthesized imidazolamines exhibited both fungal growth inhibition activity and cellular selectivity. Ned Tijdschr Geneeskd, 2003 Jan 4, 147(1), 15 - 20 {Atherosclerosis and inflammation: the role of C-reactive protein}; Ablij HC et al.; C-reactive protein (CRP) is an acute phase protein, the serum concentration of which can increase up to a 1000-fold after the onset of a stimulus . It plays a role in the aspecific immune response to bacteria and fungi and the clearance of apoptotic cell material . Inflammation plays an important role in the aetiology and pathogenesis of atherosclerosis and plaque formation . Approximately fifty percent of acute coronary syndromes are the consequence of unstable plaques rupturing, followed by thrombus formation . A characteristic of these unstable plaques is an increase in inflammatory cells (macrophages and T lymphocytes) . The serum concentration of CRP might reflect the amount of inflammation within atherosclerotic plaques and thus might provide an indirect measurement of the instability of the plaques . CRP could therefore have a predictive value for the occurrence of plaque rupture . Furthermore, there are indications that CRP itself is active in the inflammatory process . Prospective studies have shown that so-called high-sensitivity CRP (hsCRP) measurements could be used as a tool for determining the risk for acute coronary syndromes . The inflammation-inhibiting characteristics of statins and acetylsalicylic acid (especially the reduction of the hsCRP level) might contribute to reducing the risk of plaque rupture. Lin Chuang Er Bi Yan Hou Ke Za Zhi, 2000 Dec, 14(12), 573 - 5 {Comparison of the effectiveness of levofloxacin and cefuroxime for the treatment of sinusitis}; Li XP et al.; OBJECTIVE: To observe the effectiveness of levofloxacin and cefuroxime for the treatment of sinusitis in adults . METHOD: Patients with acute or chronic sinusitis were randomly assigned to receive either levofloxacin (200 mg orally once daily, 200/100 mg twice daily) or cefuroxime (250 mg orally twice daily) for 10 to 14 days . Pre- or post-treatment, the clinical and laboratory examination were done and in the end of treatment, the efficacy and safety were assessment . RESULT: The success rates after the end of treatment were 97.4% for the 76 patients who received levofloxacin and 92.8% for the 14 patients who received cefuroxime . The resolution rates of bacteria were 91.6% and 80.0%, respectively . The safety of these two groups were more high . CONCLUSION: Levofloxacin is effective for the treatment of sinusitis in adults and pay attention to the course of treatment for raising the efficacy of treatment. J Bacteriol, 2003 Feb, 185(4), 1346 - 56 Glycerol-3-phosphate acquisition in spirochetes: distribution and biological activity of glycerophosphodiester phosphodiesterase (GlpQ) among Borrelia species; Schwan TG et al.; Relapsing-fever spirochetes achieve high cell densities (>10(8)/ml) in their host's blood, while Lyme disease spirochetes do not (<10(5)/ml) . This striking contrast in pathogenicity of these two groups of bacteria suggests a fundamental difference in their ability to either exploit or survive in blood . Borrelia hermsii, a tick-borne relapsing-fever spirochete, contains orthologs to glpQ and glpT, genes that encode glycerophosphodiester phosphodiesterase (GlpQ) and glycerol-3-phosphate transporter (GlpT), respectively . In other bacteria, GlpQ hydrolyzes deacylated phospholipids to glycerol-3-phosphate (G3P) while GlpT transports G3P into the cytoplasm . Enzyme assays on 17 isolates of borreliae demonstrated GlpQ activity in relapsing-fever spirochetes but not in Lyme disease spirochetes . Southern blots demonstrated glpQ and glpT in all relapsing-fever spirochetes but not in the Lyme disease group . A Lyme disease spirochete, Borrelia burgdorferi, that was transformed with a shuttle vector containing glpTQ from B . hermsii produced active enzyme, which demonstrated the association of glpQ with the hydrolysis of phospholipids . Sequence analysis of B . hermsii identified glpF, glpK, and glpA, which encode the glycerol facilitator, glycerol kinase, and glycerol-3-phosphate dehydrogenase, respectively, all of which are present in B . burgdorferi . All spirochetes examined had gpsA, which encodes the enzyme that reduces dihydroxyacetone phosphate (DHAP) to G3P . Consequently, three pathways for the acquisition of G3P exist among borreliae: (i) hydrolysis of deacylated phospholipids, (ii) reduction of DHAP, and (iii) uptake and phosphorylation of glycerol . The unique ability of relapsing-fever spirochetes to hydrolyze phospholipids may contribute to their higher cell densities in blood than those of Lyme disease spirochetes. J Bacteriol, 2003 Feb, 185(4), 1229 - 35 QscR, a LysR-type transcriptional regulator and CbbR homolog, is involved in regulation of the serine cycle genes in Methylobacterium extorquens AM1; Kalyuzhnaya MG et al.; A new gene, qscR, encoding a LysR-type transcriptional regulator that is a homolog of CbbR, has been characterized from the facultative methylotroph Methylobacterium extorquens AM1 and shown to be the major regulator of the serine cycle, the specific C1 assimilation pathway . The qscR mutant was shown to be unable to grow on C1 compounds, and it lacked the activity of serine-glyoxylate aminotransferase, a key enzyme of the serine cycle . Activities of other serine cycle enzymes were decreased during growth on C1 compounds compared to the activities found in wild-type M . extorquens AM1 . Promoter fusion assays, as well as reverse transcription-PCR assays, have indicated that the serine cycle genes belong to three separate transcriptional units, sga-hpr-mtdA-fch, mtkA-mtkB-ppc-mcl, and gly . Gel retardation assays involving the purified QscR have demonstrated the specific binding of QscR to the DNA regions upstream of sga, mtkA, gly, and qscR . We conclude that QscR acts as a positive transcriptional regulator of most of the serine cycle enzymes and also as an autorepressor. Wei Sheng Wu Xue Bao, 2001 Jun, 41(3), 310 - 4 {Cloning and sequence analysis of the phytase phyA gene of Aspergillus niger N25}; Wang H et al.; The phyA encoding phytase of Aspergillus niger N25 was amplified by the polymerase chain reaction (PCR) with primers designed according to the sequences of the phyA in GenBank . The amplified fragment was cloned and sequenced . The results show that: the coding region is 1506 bp in size, includes a 102 bp intron, and encodes a peptide of 476 amino acid residues, in which there is a signal peptide with 19 amino acids and a mature peptide of 448 amino acids . Comparison of this sequence with the phyA of the natural A . niger NRRL3135 (GenBank Accession: M94550), the most highly secreting-phytase strain, shows that the nucleotide homology is as high as 96.746%, and the amino acid homology comes up to 97.64% . The phyA of A . niger N25 strain in this paper is appropriate to be used to construct the phytase gene-engineering bacteria. Probl Tuberk, 2002, (7), 6 - 8 {Experience with kipferon use in the treatment of patients with pulmonary tuberculosis}; Karashunskii MA et al.; Kipferon that is a combination of recombinant human (2-interferon and a complex of immunoglobulins G, M and A, was used in suppositories as an auxiliary agent in the routine chemotherapy in 36 new cases of pulmonary tuberculosis . A control group included 19 patients identical in sex, age, and the pattern of pulmonary tuberculosis . The clinical, X-ray, and laboratory indices (primarily cellular immunity) were studies before and 1 and 3 months after treatment . The beneficial effect of kipferon was manifested by a more rapid arrest of symptoms of total intoxication eliminated after 2 weeks in 39% of patients in the experimental group and only in 21% in the controls . Normalization of blood parameters occurred following a month in 58.3 and 47% of patients, respectively . Mycobacteria tuberculosis disappeared in the sputum smears following a month of treatment in 62% of those isolating bacteria in the experimental group and only in 37.5% in the controls (P > 0.1; t = 1.6) . Positive lung X-ray changes as resolved infiltration, the reduction and closure of caverns were more pronounced in the patients of the experimental group . The most characteristic change in the parameters of cellular immunity during kipferon was a short (as long as 1-1.5 months) decrease in RBT to FGA, which was noted in 47% and 6.7% of patients in the experimental and control groups, respectively (P < 0.01) and which was followed by an increase in the count of CD8+ cytotoxic lymphocytes . This is indicative of the enhancement of these mechanisms of immunity and a reduced need of enhancing or maintaining the activity of proliferative reactions of immunocompetent cells under the conditions of a favourable influence on the course of tuberculous infection. Izv Akad Nauk Ser Biol, 2002 Nov-Dec, (6), 733 - 7 {Using biological, hydrochemical and hydrological data on freshwater in Russia and neighboring countries for constructing a computerized information system}; Bulgakov NG et al.; Raw hydrobiological (population, biomass, and numbers of phytoplankton, zooplankton, zoobenthos, periphyton, and bacterioplankton species) and physicochemical indices of ecosystems are integrated into information system "Ecology of Freshwater in Russia and Neighboring Countries." This system is dedicated to environmental control by biotic indices in former USSR countries . The information system is realized as an Ecograde application relying on Microsoft Access software as well as a web site. Arch Microbiol, 2003 Jan-Feb, 179(2), 95 - 100 Epub 2002 Dec 14. Novel carotenoid glucoside esters from alkaliphilic heliobacteria; Takaichi S et al.; Pigments of three species of alkaliphilic heliobacteria of the genus Heliorestis, H . daurensis, H . baculata and an undescribed species Heliorestis strain HH, were identified using spectroscopic methods . In these species, bacteriochlorophyll g esterified with farnesol was present, as for other heliobacteria . The carotenoids consisted of 4,4'-diaponeurosporene, also found in other heliobacteria, plus the novel pigments OH-diaponeurosporene glucoside esters (C16:0 and C16:1) . In addition, trace amounts of biosynthetic intermediates, OH-diaponeurosporene and OH-diaponeurosporene glucoside, were found . Trace amounts of a carotenoid with 20 carbon atoms, 8,8'-diapo-zeta-carotene, were also found in these species as well as in the non-alkaliphilic heliobacteria . The non-alkaliphilic species Heliophilum fasciatum also contained trace amounts of the two OH-diaponeurosporene glucoside esters . The results are used to predict the pathway of carotenoid biosynthesis in heliobacteria. Nucleic Acids Res, 2003 Feb 1, 31(3), 1038 - 44 Synapsis and strand exchange in the resolution and DNA inversion reactions catalysed by the beta recombinase; Canosa I et al.; In the presence of a sequence-independent chromatin-associated protein, such as Hbsu or HMGB, the beta recombinase catalyses resolution between two directly oriented recombination sites (six sites) and both resolution and DNA inversion between two inversely oriented six sites . Assembly of the synaptic complex requires binding of the beta recombinase to the six sites and the presence of Hbsu . Whether resolution or inversion will take place depends on the relative orientation of the two six sites, the level of DNA supercoiling and the amounts of Hbsu . In this work, the topologies of the products of the resolution and inversion reactions were analysed . The resolution reaction generated mainly singly catenated DNA circles, while DNA inversion gave rise to unknotted circles and small amounts of DNA molecules containing 3- or 5-noded knots . In spite of the distinctive features of the beta system, the topology of synapsis and strand exchange during the resolution reaction is similar to that of Tn3 and gammadelta resolvases . The ability of the beta recombinase to catalyse both inversion and resolution reactions probably reflects different possible architectures of the synaptic complex, which to a large extent depends on Hbsu. Exp Gerontol, 2002 Dec, 37(12), 1359 - 69 No reduction of metabolic rate in food restricted Caenorhabditis elegans; Houthoofd K et al.; Dietary restriction (DR) is the most consistent means of extending life span throughout the animal kingdom . Multiple mechanisms by which DR may act have been proposed but none are clearly predominant . We asked whether metabolic rate and stress resistance is altered in Caenorhabditis elegans in response to DR . DR was imposed in two complementary ways: by growing wild-type worms in liquid medium supplemented with reduced concentrations of bacteria and by using eat-2 mutants, which have a feeding defect . Metabolic rate was not reduced when we fed wild-type worms reduced food and was up-regulated in the eat-2 mutants in liquid culture, as assessed by oxygen consumption rate and heat production . The specific activity levels of the antioxidant enzymes superoxide dismutase (SOD) and catalase showed small increases when we reduced food in wild-type worms, but restricted worms acquired no elevated protection against paraquat and hydrogen peroxide . eat-2 mutants showed elevated specific activities of SOD and catalase relative to wild type in liquid culture . These results indicate that the effects imparted by DR and the eat-2 mutation are not identical, and they contradict, at least in C . elegans, the widespread belief that CR acts by lowering the rate of metabolism. Life Sci, 2003 Feb 28, 72(15), 1745 - 56 Evolutional study on acetylcholine expression; Horiuchi Y et al.; Acetylcholine (ACh) is a well-known neurotransmitter in the cholinergic nervous systems of vertebrates and insects; however, there is only indirect evidence for its presence in lower invertebrates, such as plants and fungi . We therefore investigated the expression of ACh in invertebrates (sea squirt, sea urchin, trepang, squid, abalone, nereis, sea anemone, coral and sponge), plants (arabidopsis, eggplant, bamboo shoot, cedar, hinoki, pine, podcarp, fern, horsetail and moss), fungi (yeast and mushroom) and bacteria by assaying ACh content and synthesis, focusing on the presence of two synthetic enzymes, choline acetyltransferase (ChAT) and carnitine acetyltransferase (CarAT) . Using a specific radioimmunoassay, ACh was detected in all samples tested . The levels varied considerably, however, with the upper portion of bamboo shoots having the highest content (2.9 micromol/g) . ACh synthesis was also detected in all samples tested; moreover, the activity in most samples from the animal kingdom, as well as bamboo shoots and the stem of the shiitake mushroom, were sensitive to both ChAT and CarAT inhibitors . Levels of ACh synthesis were lower in samples from other plants, fungi and bacteria and were insensitive to ChAT and CarAT inhibitors . These findings demonstrate the presence of ACh and ACh-synthesizing activity in evolutionally primitive life as well as in more complex multicellular organisms . In the context of the recent discovery of non-neuronal ACh in various mammalian species, these findings suggest that ACh been expressed in organisms from the beginning of life, functioning as a local mediator as well as a neurotransmitter . J Gastrointest Surg, 2003 Jan, 7(1), 68 - 76 Helicobacter pylori induces apoptosis in Barrett's-derived esophageal adenocarcinoma cells; Jones AD et al.; Helicobacter pylori may protect against the development of dysplasia in Barrett's epithelium of patients with gastroesophageal reflux disease . The aim of this study was to determine whether H . pylori preferentially induces apoptosis in Barrett's-derived cancer cells compared to normal cells . A Barrett's-derived adenocarcinoma cell line (OE33) was grown . H . pylori wild-type, isogenic vacA-, cagA(-), and picB-/cagE- mutant strains were grown on agar plates . Intact or sonicated bacteria were used to treat normal and OE33 cells for 24 hours, and Hoechst dye binding was performed to measure apoptosis . FAS protein expression was determined by Western immunoblotting . OE33 cells treated with intact H . pylori wild-type strains produced significant (P < 0.05) dose-dependent increases in apoptosis compared to normal esophageal cells . H . pylori wild-type and vacA- isogenic strains were more effective than cagA- and picB-/cage- isogenic strains in inducing apoptosis in OE33 cells . In OE33 cells, H . pylori sonicates produced lower levels of apoptosis than intact bacteria . Wild-type H . pylori strains increased Fas protein expression in OE33 cells at 18 hours . H . pylori induced apoptosis at a higher rate in the Barrett's-derived human esophageal adenocarcinoma cells than in normal esophageal cells . The H . pylori-induced apoptosis was primarily dependent on intact bacteria and the presence of the cagA and picB/cagE gene products . H . pylori-induced apoptosis may involve the Fas-caspase cascade . Histopathology, 2003 Feb, 42(2), 173 - 85 Splenic marginal zone atrophy and progressive CD8+ T-cell lymphocytosis in HIV infection: a study of adult post-mortem spleens from Côte d'Ivoire; Wilkins BS et al.; AIMS: Progressive changes have been reported in lymph nodes in HIV infection, but few accounts describe altered splenic histology at different stages of the disease . Investigation of splenic changes accompanying the progressive CD4+ T-cell depletion that occurs in HIV infection could shed light on normal immunological interactions in this organ . Therefore, we assessed the amount and distribution of lymphoid tissue in spleens from adults with documented early or advanced HIV disease . METHODS AND RESULTS: Immunohistochemistry was used to study splenic tissue collected in an extensive autopsy survey of HIV+ adults in West Africa . Compared with post-mortem spleens from HIV- West African adults and control UK spleens, those from HIV-infected patients showed severe atrophy of white pulp B- and T-cell compartments . In early and advanced HIV disease, marginal zone atrophy was significant . Peri-arteriolar lymphoid sheaths contained increased numbers of CD8+/CD45RO+ T-cells in advanced HIV disease . In red pulp, early and advanced cases showed a lymphocytosis of CD8+/CD45RO- T-lymphocytes . CONCLUSIONS: Atrophic changes were more extreme in advanced than early HIV infection . Reduced marginal zone function possibly explains the known predisposition of HIV+ patients to infection by encapsulated bacteria . Possible immunological consequences of these CD8+/CD45RO+ (peri-arteriolar lymphoid sheaths) and CD8+/CD45RO- (red pulp) responses deserve further study . Comparison of West African and UK control spleens indicated that there were no major ethnic differences in spleen structure to prevent extrapolation of our results to European adults. Environ Microbiol, 2003 Feb, 5(2), 116 - 26 Intracellular symbionts of sharpshooters (Insecta: Hemiptera: Cicadellinae) form a distinct clade with a small genome; Moran NA et al.; The leafhoppers (Insecta: Hemiptera: Cicadellidae) are the most species-rich group of invertebrates in which intracellular symbionts are usual . Here we present the first molecular characterization of bacteriome-associates in the leafhoppers, with focus on the subfamily Cicadellinae (sharpshooters) . Phylogenetic analyses of 16S rDNA sequences from intracellular symbionts residing in the bacteriomes of five host species indicate that these symbionts form a well-defined clade within the gamma-3 Proteobacteria, consistent with an ancient colonization and strict vertical transmission . More extensive gene sequence information is reported for the symbiont of Homalodisca coagulata (Say) . The genome size, as determined by pulsed field gel electrophoresis, is approximately 680 kb . This finding, when combined with published results for symbionts of aphids, ants, psyllids and tsetse flies, adds to an emerging pattern which suggests that bacteriome associates often descend from ancient infections by gamma Proteobacteria, and that these lineages have undergone pronounced genome reduction . A new genus and species name, 'Candidatus Baumannia cicadellinicola' (sp . nov.) is proposed for this newly characterized clade of symbiotic bacteria. Sheng Wu Yi Xue Gong Cheng Xue Za Zhi, 2000 Jun, 17(2), 231 - 2 {An experiment on disinfection using high power microwave}; Xi X et al.; This experimental disinfection was intended to examine the effect of high power microwave on bacteria, and the putative nonthermal effect . The bacteria of the experimental groups were irradiated by high power microwave in different intensities and time and were compared with the controls--bacteria heated only . The principles of disinfection by means of high power microwave were studied . The results showed that the temperature did not change after the dry bacteria were irradiated by high power microwave in different intensities and time, but the number of bacteria alive decreased significantly(P < 0.05) . The high power microwave's effect of disinfection was greater in the wet bacteria group than in the dry bacteria group and the bacteria heated group . These data indicate that the disinfection effect of high power microwave is higher than that of heating . High power microwave can kill dry bacteria, and it has the nonthermal effect in addition to its thermal effect. Ying Yong Sheng Tai Xue Bao, 2002 Oct, 13(10), 1344 - 8 {Effects of elevated atmospheric CO2 on the exchange of trace gases between ecosystems and the atmosphere}; Xu Z et al.; The latest researches on the effects of elevated atmospheric CO2 on the exchanges of trace gases (e.g., CO2, CH4 and N2O) between the atmosphere and ecosystems were reviewed . The techniques and methods involved in the researches were introduced firstly . Then the review mainly focused upon the results from those studies using the open-top-chamber (OTC) methods and the free-air carbon dioxide enrichment (FACE) system . Generally, elevated atmospheric CO2 may stimulate biomass accumulation, and enlarge C/N ratio in plant tissue so as to reduce the decomposition of organic matter . This action could increase CO2 sequestration in terrestrial ecosystems . Elevated atmospheric CO2 could impact on methanogenic bacteria and CH4 emissions . An increase in CH4 emissions from wetland may appear . The argument among the responses of N2O emissions to elevated CO2, however, was inconsistent . So far, no study on other trace gases was reported . More efforts should be taken in the research on the effects of elevated atmospheric CO2 on the exchange of trace gases. Wei Sheng Wu Xue Bao, 2002 Apr, 42(2), 214 - 9 {Studies on hydantoinase producing conditions of Arthrobacter K1108}; Zhang W et al.; The hydantoinase-producing conditions from strain Arthrobacter K1108 were investigated . It is shown that the hydantoinase in the strain is intracellular and inducible . Its optimal inducer is 5-benzylhydantoin, while 5-indolylmethylhydantoin and 5-phenylhydantoin cannot induce the production of hydantoinase in K1108 . A gratuitous inducer was designed, with which the hydantoinase production is 343% as much as that with 5-benzylhydantoin . The media for culturing the bacteria were screened and optimized . Under optimal conditions, the specific activity of K1108 cells reached 10.8 U/mL. Wei Sheng Wu Xue Bao, 2002 Feb, 42(1), 33 - 9 {The cloning and functional analysis of Sinorhizobium fredii 042BS regulatory modulation genes}; Zhang H et al.; The total DNA of Sinorhizobium fredii 042BS was digested by EcoRI for Southern blotting with probes of nodD1 and nodD2 from S . fredii USDA257 . The 3 kb positive band hybrided with nodD1 probe and 6 kb positive band with nodD2 probe were found, respectively . Partial gene library were constructed using pUC18 as vector, and the clones with the nodD1 and nodD2 genes were obtained . The sequence of nodD1 and nodD2 of 042B showed that they are highly homologous with nodD1 and nodD2 of S . fredii . The fragment with nodD1 was cloned into the vector pBBRIMCS-5 and introduced into R . leguminosarum bv . viciae LPR5054 to study the function of the nodD1 . The results showed that nodD1 of 042B can be induced by genistein and luteolin secreted by the seedlings of soybean and alfalfa respectively. Phytother Res, 2003 Jan, 17(1), 19 - 25 Seasonal variation of the lipoidal matters and hypolipidaemic activity of the red alga Corallina officinalis L; Awad NE et al.; The lipoidal matters of Corallina officinalis L . showed a seasonal nonsignificant quantitative variation . However, the fatty acids revealed a relative increase in the summer and winter, while unsaponifiable matter exhibited a slight increase in the spring . The GC/MS analysis of saponifiable and unsaponifiable matter of the algal samples collected in different seasons revealed that samples collected in the spring contained a low cholesterol content and high steroidal compounds as well as high polyunsaturated fatty acids . The alcohol extract, hexane extract and fatty acid fraction of this algal sample exhibited a significant hypolipidaemic activity . Also, two biologically active fractions of hydrocarbons were isolated by CC technique from the hexane fraction of C . officinalis L . and identified by GC/MS . Am J Trop Med Hyg, 2003 Jan, 68(1), 92 - 6 Subcellular localization of rickettsial invasion protein, InvA; Gaywee J et al.; To understand further the molecular basis of rickettsial host cell invasion, Rickettsia prowazekii invasion gene homolog (invA) has been characterized . Our previous experiments have shown that InvA is an Ap5A pyrophosphatase, a member of the Nudix hydrolase family, which is up-regulated during the internalization, early growth phase, and exit steps during rickettsial mammalian cell infection . In addition to the molecular characterization, subcellular localization of InvA was investigated . InvA-specific antibodies were raised in mice and used for immunoelectron microscopy . The generated antibodies were shown to recognize InvA and by immunogold labeling showed InvA in the cytoplasm of rickettsiae . A cytoplasmic location for InvA would allow for a rapid response to any internal substance and efficient functioning in hydrolysis of toxic metabolic by-products that are accumulated in the rickettsial cytoplasm during host cell invasion . Protecting bacteria from a hazardous environment could enhance their viability and allow them to remain metabolically active, which is a necessary step for the rickettsial obligate intracellular lifestyle. Respir Med, 2003 Jan, 97(1), 46 - 50 Effects by 8-bromo-cyclicAMP on basal and organic dust-induced release of interleukin-6 and interleukin-8 in A549 human airway epithelial cells; Burvall K et al.; Inhalation of organic dust from a swine-confinement building leads to an intense inflammatory reaction with an increased number of inflammatory cells and mediators in the upper and lower respiratory tract of previously unexposed subjects . In vitro the dust induces cytokine release from epithelial cells and alveolar macrophages . It is known that intracellular cyclic AMP (cAMP) contributes to the regulation of inflammatory responses . We therefore investigated whether 8-Bromo-cAMP, a cell membrane-permeable cAMP analogue, would influence release of the cytokines interleukin-6 (IL-6) and IL-8 in a human airway epithelial cell line, A549, exposed to a suspension of the organic dust, and to a supernatant prepared by centrifugation (at low g-force) of a suspension of dust . The large particulate matter was thus sedimented, leaving bacteria, whole and cell wall constituents in the supernatant . Cytokine release was measured with enzyme-linked immunosorbent assay (ELISA) . The cytokine release induced by a supernatant was 23% (IL-6) and 27% (IL-8) of the release induced by a dust suspension . 8-Bromo-cAMP (1 mM) doubled basal IL-6 release and IL-6 release induced by a dust supernatant (P<0.01), and increased IL-6 release induced by a dust suspension by 19% (P<0.05) . 8-Bromo-cAMP did not affect basal IL-8 release, partially inhibited (28%) the release of IL-8 induced by a dust suspension (P<0.01), but increased IL-8 release induced by a dust supernatant by 13% (P<0.05) . In summary, expression of the cytokines IL-6 and IL-8 is differentially regulated by 8-Bromo-cAMP, both with regard to basal and dust-induced release . The results indicate that 8-Bromo-cAMP attenuated IL-8 release by affecting signaling transductions induced by the particulate fraction. J Am Dent Assoc, 2003 Jan, 134(1), 87 - 95 Minimally invasive dentistry; Murdoch-Kinch CA et al.; BACKGROUND: During the past few decades, scientific developments in cariology, dental materials and diagnostic systems have changed dentistry's approach to diagnosis and management of dental caries . The authors summarize these developments . OVERVIEW: Dental adhesives and restorative materials, new understanding of the caries process and remineralization, and changes in caries prevlance have catalyzed the evolution in caries management from G.V . Black's "extension for prevention" to "minimally invasive." The authors describe the scientific basis for early diagnosis; a modified classification of caries based on site and size of lesion remineralization; reduction of cariogenic bacteria; and minimally invasive cavity preparation design, techniques and material selection . CONCLUSIONS AND PRACTICE IMPLICATIONS: Minimally invasive dentistry is based on advances in science . Emerging technologies will facilitate evolution to primary prevention of caries, though technical, cultural and economic obstacles to full implementation in clinical practice now exist. Faraday Discuss, 2003, 122, 65 - 77; discussion 79-88 The frontiers of time-resolved macromolecular crystallography: movies and chirped X-ray pulses; Moffat K; Three important frontiers of ultrafast time-resolved macromolecular crystallography are presented: extension of this technique to other biological systems; further developments in the elucidation of mechanism through the analysis of time-dependent movies to extract the underlying, time-independent, intermediate structures; and enhanced time resolution . The last is intimately linked with the nature of the pump-probe experiment itself, with the sources of random and, particularly, systematic experimental error, and with the factors that contribute to overall time resolution . All experiments to date have utilized the unchirped X-ray pulses that are emitted by synchrotron sources . Chirped pulses offer certain advantages for ultrafast X-ray experiments such as those based on Laue diffraction . An energy-chirped pulse maps photon energy into time; a Laue diffraction experiment maps photon energy into detector space . Hence, a Laue experiment with an energy-chirped pulse maps time into space . The proposed sub-picosecond photon source could provide an excellent source of intense, chirped hard X-rays for such experiments. Acta Crystallogr D Biol Crystallogr, 2003 Feb, 59(Pt 2), 203 - 13 Epub 2003 Jan 23. Lysosomal cysteine proteases (cathepsins): promising drug targets; Turk D et al.; Papain-like lysosomal cysteine proteases are processive and digestive enzymes expressed in organisms from bacteria to humans . Their ubiquity alone makes them potential drug targets, with the assumption that appropriate specificities may be achieved . These enzymes have rather short active-site clefts, comprising three well defined substrate-binding subsites (S2, S1 and S1') and additionally have comparatively broad binding areas (S4, S3, S2', S3') . This geometry distinguishes them from other protease classes, such as serine and aspartic proteases, with six and eight substrate-binding sites, respectively . Exopeptidases (cathepsins B, C, H and X), in contrast to endopeptidases (such as cathepsins L, S, V and F), possess structural features that facilitate binding of N- and C-terminal groups of substrates in the active-site cleft . Other than a clear preference for free chain termini in the case of exopeptidases, the substrate-binding sites exhibit no strict specificities . Instead, their subsite preferences arise more from specific exclusions of substrate type . This presents a challenge for the design of inhibitors to target a specific cathepsin: only the cumulative effect of an assembly of inhibitor fragments can produce the desired result . The small number of papain-like lysosomal cysteine proteases (11 human enzymes are known) and the small number of substrate-binding sites calls for a in