Adv Dermatol, 1995, 10, 77 - 96; discussion 97 Wound healing; Waldorf H et al.; Wound healing is a dynamic biologic process of repairing insults to the integumentary system . It is commonly divided into three phases: inflammatory, proliferative, and maturation . Each phase has unique cellular and substance constituents without which it cannot progress normally . A large variety of factors may influence any part of wound healing, including local factors such as bacteria, oxygen tension, and bleeding, and systemic factors such as the mental and physical health of the patient . There are also extrinsic factors that can be influenced by the caretakers of the wound to enhance wound healing . Areas of intervention include using antiseptic technique when one is dealing with the wound, using good surgical technique, choosing the appropriate wounding method and repair for the individual patient, and using antibiotics and special wound dressings . Modern science and technology are giving us new insights into wound healing and leading us to exciting new ways of influencing it, including the topical use of growth factors, artificial skins, cultured epithelium with and without dermal components, and electrical stimulation . The future of wound healing holds a better understanding of the complexities of the physiologic events that occur and a translation of that into a biologically active and interactive wound care. Avian Dis, 1995 Jan-Mar, 39(1), 183 - 90 Enteritis in turkeys associated with an unusual flagellated protozoan (Cochlosoma anatis); Cooper GL et al.; Outbreaks of enteritis associated with an unusual flagellated protozoan occurred in six California turkey flocks during the summer of 1992 . Certain morphological and ultrastructural details of the parasite, which resembles Cochlosoma anatis, are illustrated with scanning and transmission electron micrographs . The flagellate attached to the intestinal mucosa by means of a sucker-like apparatus, and circular impressions of the sucker were created on the surface epithelium . Histological lesions were characterized by blunting and fusion of villi; cellular infiltration of the lamina propria with lymphocytes, plasma cells, histiocytes, and heterophils; and increased numbers of mitotic figures in crypt epithelium. Curr Genet, 1995 Jan, 27(2), 142 - 9 Sequence analysis of the Aspergillus nidulans pectate lyase pelA gene and evidence for binding of promoter regions to CREA, a regulator of carbon catabolite repression; Ho MC et al.; The nucleic acid and deduced amino-acid sequences of the pectate lyase gene (pelA) from Aspergillus nidulans are presented . The pelA gene contains two short introns, 68 and 49 bp in length, and encodes a peptide of 326 amino acids . Five transcriptional start sites are clustered between 65 and 79 bp upstream of the start codon as determined by primer extension . Comparison of the amino-acid sequences of pectate or pectin lyases from bacteria, fungi and plants revealed less than 30% overall identity . However, five regions within these enzymes, in particular domains associated with the active site, are highly conserved with amino-acid similarities greater than 50% . Phylogenetic analysis using the principle of parsimony (PAUP 3.1.1) showed that pelA is most closely related to pectate lyases from plants rather than pectin lyases from other fungi . Previously, pelA was shown to be induced by polygalacturonic acid and repressed in the presence of preferred carbon sources, such as glucose . Gel mobility shift analysis indicates that a PstI-SphI fragment from the pelA promoter binds to a fusion protein composed of the N-terminal part of CREA, a protein involved in carbon catabolite repression, and glutathione-S-transferase . This result suggests CREA may contribute to the regulation of pelA expression. Microb Pathog, 1995 Jan, 18(1), 29 - 36 Molecular cloning of an Actinobacillus pleuropneumoniae outer membrane lipoprotein (OmlA) from serotype 5a; Ito H et al.; The gene encoding an outer membrane lipoprotein (OmIA) was cloned from Actinobacillus pleuropneumoniae strain NG-8 (serotype 5a) . The deduced amino acid sequence of OmIA from strain NG-8 showed 61% identity to the OmIA from serotype 1 strain, which confers protective immunity to pigs . Southern blot analysis showed the presence of a sequence highly homologous to the omIA gene of strain NG-8 in strains of serotype 5a, 5b and 10 . A specific serum against OmIA of NG-8 also detected a homologous protein in the strains of these serotypes . These data shows the presence of antigenic variability among A . pleuropneumoniae OmIA proteins. J Vet Diagn Invest, 1995 Jan, 7(1), 98 - 101 Immunohistochemical detection of Tritrichomonas foetus in formalin-fixed, paraffin-embedded sections of bovine placenta and fetal lung; Rhyan JC et al.; An immunohistochemical technique using a monoclonal antibody was evaluated as a diagnostic tool to specifically label Tritrichomonas foetus in formalin-fixed, paraffin-embedded sections of placenta and fetal lung from bovine abortions . Trichomonads were demonstrated in tissues from each of 12 abortions due to T . foetus and none of 15 abortions due to other or unidentified causes . Moderate to marked background staining occurred only in severely autolyzed tissues from T . foetus-infected fetuses . The antibody faintly labeled 1 of 3 other species of trichomonads (Trichomonas gallinae) but did not label other protozoa, bacteria, or fungi tested. Protein Eng, 1995 Jan, 8(1), 1 - 4 DNA recognition by a beta-sheet; Suzuki M; DNA recognition by a beta-sheet is discussed in the light of crystal structures of the MetJ and Arc repressors . The DNA binding geometry of a beta-sheet can be understood in terms of (i) close fitting of the two surfaces and (ii) matching of residue and base positions . A beta-sheet is not entirely flat but has a curvature . A beta-sheet of the Met-Arc family faces the DNA major groove with its convex surface; the local DNA major groove is deepest at the centre . The beta-sheet follows 6 bp; every two residues face the DNA and the first and fifth residues, which are separated by 13.2 A, bind, respectively, to the third and sixth bases, which are separated by 13.5 A, on the same DNA strand. Surg Today, 1995, 25(1), 49 - 54 Occurrence of intraperitoneal septic complications after hepatic resections between 1985 and 1990; Matsumata T et al.; In this study, the risk factors related to intraperitoneal septic complications occurring after hepatectomy (IPSCH) as well as the effect of various perioperative variables on the outcome of IPSCH between 1985 and 1990 were analyzed . Twenty-one of 211 patients (10.0%) developed IPSCH . The findings in the patients with IPSCH were compared with those in 190 patients without IPSCH . The significant variables associated with the development of IPSCH included a high incidence of accompanying chronic renal failure (14.3% vs 2.1%), a larger blood loss during surgery (2,130 vs 1,340 ml) as well as a greater amount of intraoperative blood replacement (1,130 vs 570 ml), and a greater weight of the resected liver (367 vs 233 g) . IPSCH occurred in 10 of 12 patients who had postoperative bile leakage . Eighteen patients (85.7%) with IPSCH were discharged from the hospital after non-operative management; however, the hospital death rate (14.3% vs 1.1%) was significantly higher in patients with IPSCH . This review suggests that the incidence of IPSCH has not decreased recently . Thus, to prevent IPSCH, at least following bile leakage, it is necessary to perform a careful division of the liver parenchyma followed by a bile leakage test, and when this complication occurs unexpectedly in patients who have a good functional reserve of the remnant liver, IPSCH can be effectively drained percutaneously under ultrasound guidance. Ann Pharm Fr, 1995, 53(1), 29 - 37 {Iminodimethylation, a method for pharmacomodulation in pyrimido-{3,4-a}-s-triazine series}; Menager S et al.; Primary amines react with two formaldehydes and compounds presenting two mobile hydrogen atoms, this reaction can be called iminodimethylation . This reaction can be used in order to perform a pharmacomodulation in the pyrimido {3,4-a}-s-triazine series . Against Epidermophyton floccosum, the activity is better when nitrogen 7 is not substituted, when the heroatom in position 2 is 0 instead of S and when an aromatic nucleus is directly linked to the nitrogen atom in position 3. J Med Microbiol, 1995 Jan, 42(1), 48 - 52 In-vitro hepatotoxic factor in Helicobacter hepaticus, H . pylori and other Helicobacter species; Taylor NS et al.; Several inbred strains of mice in closed breeding colonies were found to have spiral-shaped bacteria associated with active, chronic hepatitis . A new species of Helicobacter, H . hepaticus, was isolated from the infected livers of some strains of mice . Other strains of mice were colonised with H . hepaticus in the caecum and colon, but not the liver . Filtersterilised supernatant fluid from five strains of H . hepaticus was tested in a mouse liver cell line (ATCC no . CCL 9.1) for cytotoxic activity . All strains produced a toxic factor causing morphological changes in the cells at dilutions up to 1 in 1000 . Toxicity was observed after exposure to the supernatant fluid for 48-72 h . Other Helicobacter spp . that also produced the cytopathic effect (CPE) in the liver cell line were H . felis, H . acinonyx, H . pylori and one strain of H . mustelae . "Helicobacter rappini" and H . muridarum did not cause CPE in the liver cells . The soluble factor was stable at 4 degrees C for up to 3 months . It was also stable at 56 degrees C for 30 min, but was inactivated by boiling for 15 min . It was inactivated by incubation with trypsin . A partially purified preparation of the cytotoxin had a mol . wt of c . 100,000 and did not have urease activity . The cytotoxin produced by H . hepaticus did not cause vacuole formation in HeLa cells. J Reprod Immunol, 1995 Jan, 28(1), 1 - 13 IgG asymmetric anti-ovalbumin antibodies synthesized by virgin and pregnant rats; Gentile T et al.; A study on the synthesis of asymmetrical IgG molecules 'with no specific activity' and with anti-ovalbumin activity was carried out both in virgin rats and in rats inoculated with ovalbumin and made pregnant by syngeneic and allogeneic males . Before pregnancy, female rats synthesize about 23% of asymmetrical IgG molecules, when the level of these molecules is assessed in total IgG, in anti-ovalbumin IgG and in the supernatant from the adsorption of anti-ovalbumin antibodies . On the other hand, anti-ovalbumin antibodies isolated are predominantly of the symmetrical IgG type; they are also precipitants and effectors of the biological mechanisms that the host operates to preserve pathogenic antigens (bacteria, parasites) . In rats pregnant by syngeneic and allogeneic males, the ratio asymmetric/symmetric IgG molecules increases, and the anti-ovalbumin antibodies are mainly of the asymmetrical IgG type, which aid antigen-blocking . Similar results are found in virgin rats, immunized with ovalbumin and intraperitoneally transferred simultaneously with supernatants of placental cultures . These results suggest that, during pregnancy, there is an increase of the IgG asymmetric/symmetric molecule ratio, produced by placental factors, whatever the immunogen specificity may be . Speculations about this fact are presented. J Hosp Infect, 1995 Jan, 29(1), 65 - 8 Stethoscope contamination in the neonatal intensive care unit; Wright IM et al.; The level of contamination of stethoscopes used in a neonatal intensive care unit was studied, along with the practices used for cleaning these items . A policy of alcohol cleaning was introduced and the effect of this change on the level of bacterial growth was observed after a six-week period . It was found that 71% of stethoscopes had a significant bacterial growth and that this was reduced to 30% after the cleaning procedure change (P < 0.05) . Stethoscopes and other equipment are a potential source of nosocomial infection on the neonatal intensive care unit. Dev Genet, 1995, 16(2), 179 - 89 Genomic organization of a mouse glyceraldehyde 3-phosphate dehydrogenase gene (Gapd-s) expressed in post-meiotic spermatogenic cells; Welch JE et al.; The Gapd-s gene encodes an isoform of the glyceraldehyde 3-phosphate dehydrogenase enzyme expressed only in post-meiotic spermatogenic cells . Two clones containing the Gapd-s gene were isolated from a mouse genomic library . Sequencing and restriction enzyme analysis demonstrated that this single-copy gene contains 11 exons and spans 9596 base pairs . The locations of Gapd-s exons and introns are conserved when compared to the corresponding portions of the chicken and human somatic Gapd genes . The promoter region contains no TATA box, although there is a potential SP1 recognition site within exon 1 . Like other TATA-less genes, primer extension analysis reveals some heterogeneity in the site of transcription initiation with Gapd-s transcripts initiating from three discrete sites . Northern analysis demonstrated that a 1.5-kb Gapd-s mRNA is expressed in the testis in at least three mammalian orders, indicating that the Gapd-s gene appeared early in mammalian evolution . Using GAPD-deficient bacteria, mouse GAPD-S was shown to be capable of functioning as a glycolytic enzyme . Since GAPD has been proposed to be a key enzyme regulating glycolysis in spermatogenic cells, GAPD-S may represent a potential target for toxicological or contraceptive agents affecting fertility by interfering with glycolysis. Crit Rev Biotechnol, 1995, 15(1), 13 - 39 Production and secretion of proteins by streptomycetes; Gilbert M et al.; Streptomycetes produce a large number of extracellular enzymes as part of their saprophytic mode of life . Their ability to synthesize enzymes as products of their primary metabolism could lead to the production of many proteins of industrial importance . The development of high-yielding expression systems for both homologous and heterologous gene products is of considerable interest . In this article, we review the current knowledge on the various factors that affect the production and secretion of proteins by streptomycetes and try to evaluate the suitability of these bacteria for the large-scale production of proteins of industrial importance. Zentralbl Bakteriol, 1995 Jan, 282(1), 50 - 3 Adhesion to and invasion of HeLa cells by Helicobacter pylori; Rautelin H et al.; Eight clinical isolates and two reference strains of Helicobacter pylori were studied with regard to their interactions with HeLa cells . All the isolates adhered poorly to HeLa cells and the number of invasive bacteria was very low . No correlation was found between the adherence and invasiveness of the isolates on one hand, and the corresponding patients having ulcer or non-ulcer disease, or the ability of the strains to produce cytotoxin and to induce an oxidative burst of human polymorphonuclear leukocytes without opsonins, on the other . These results indicate that invasion of epithelial cells would play no important role in the pathogenesis of infections caused by H . pylori. Zentralbl Bakteriol, 1995 Jan, 282(1), 18 - 23 Investigations of culture and properties of Afipia spp; Muller HE; The culture conditions of Afipia felis, A . broomeae, A . clevelandensis and three unnamed Afipia genospecies were investigated on BCY agar supplemented with different substances known as growth factors of Legionella spp . and, furthermore, with sodium chloride and other salts . The organisms were found to be susceptible to a certain degree to byproducts of the autoclaving which are scavenged by activated by charcoal . Growth was weakly enhanced by ferric pyrophosphate, cystein.HCl, and alpha-ketoglutarate . These substances are no obligatory growth factors . The optimal pH value was about 6.8 . Afipia spp . showed a strong susceptibility to NaCl and other salts . They possess phosphatase, phosphoamidase, phosphodiesterase, a weak sulfatase, glycine aminopeptidase, and L-lysine aminopeptidase . The strains differed with regard to other proteases and aminopeptidases . The decimal reduction times of A . felis at 55 degrees C and 60 degrees C were 11 min, < 1 min, respectively. Vet Parasitol, 1995 Jan, 56(1-3), 207 - 23 A comparison of inflammatory exudates released from myiasis wounds on sheep bred for resistance or susceptibility to Lucilia cuprina; O'Meara TJ et al.; Sheep bred for resistance (R) or susceptibility (S) to fleece rot and myiasis (blowfly strike) were experimentally infected with L . cuprina larvae . Exudates released from the wound site were collected during the infection at 6, 12, 18 and 24 h . The exudates were separated using two-dimensional gel electrophoresis, and proteins were silver stained and identified by immunoblotting with specific antibody and by their isoelectric points and molecular weights . Comparisons of exudate composition were made over time and between R and S sheep . Between 6 and 12 h post larval implantation the exudate was rich in IgG and fibrinogen, which is before extensive tissue damage and suggests that the exudate is not simply tissue haemorrhage but the result of an inflammatory response by the sheep to L . cuprina . The exudate grew in complexity between 12 and 18 h and contained a maximum of 74 distinct peptide spots by 24 h . Exudate from wounds on resistant sheep contained many more peptides in the first 12 h of infection, suggesting a more rapid inflammatory response . The source of proteins from the exudate remains speculative; it appears to be composed of many acute-phase proteins, large amounts of immunoglobulin G and proportionally low levels of serum albumin . Exudate composition is likely to be influenced by the local synthesis of acute-phase proteins and perhaps immunoglobulins, selective transport to the infection site and also enzymic degradation by L . cuprina larval enzymes . The more rapid exudation of acute-phase and serum proteins at infection sites on R sheep may allow the inhibition of the establishment of fleece rot bacteria or L . cuprina larvae under natural challenge. Acta Clin Belg, 1995, 50(1), 4 - 8 {Treatment of tuberculosis with a fixed combination antitubercular drug in a marginal population in Brussels}; Alame T et al.; Tuberculosis is a world-wide persistent problem . Treatment is usually successful when the bacteria are drug-susceptible and patient compliance ensured . We report our results of treatment of high-risk patients without social security coverage in Brussels . Double and triple fixed antituberculous agents in combined tablets were used . Cure was obtained without drug toxicity in all patients who completed therapy . Only 19.5% of patients failed to complete therapy. J Oral Pathol Med, 1995 Jan, 24(1), 14 - 7 Cytomegalovirus (CMV) and Helicobacter pylori (HP) found in oral mucosal ulcers; Leimola-Virtanen R et al.; The possible involvement of Cytomegalovirus (CMV) and Helicobacter pylori (HP) in oral mucosal ulcers is suggested by their role in the development of ulceration at other mucosal sites of the gastrointestinal tract . A series of 29 incisional biopsies from 29 consecutive and apparently immunocompetent patients attending the clinic for oral ulceration were examined by routine histopathology as well as by in situ hybridisation (ISH) with biotinylated CMV and HP DNA probes . In 14/29 biopsies, Giemsa staining disclosed spiral bacteria . Six (20.7%) of these 14 Giemsa-positive samples showed HP DNA on ISH and 3 ulcers (10.3%) contained CMV DNA . In none of the specimens were CMV and HP detected simultaneously . Two of the ulcers containing CMV DNA were found on the labial mucosa and one on the posterior palatal mucosa, whereas all HP DNA-positive ulcers were located on the buccal mucosa . The results indicate that CMV and HP DNA can be found in separate oral mucosal ulcers in apparently immunocompetent adults. J Periodontal Res, 1995 Jan, 30(1), 23 - 33 Evidence of a direct relationship between neutrophil collagenase activity and periodontal tissue destruction in vivo: role of active enzyme in human periodontitis; Lee W et al.; To assess the temporal relationship between periodontal tissue destruction and the activity of collagenase, exudate from inflamed periodontal tissues was collected and latent and active collagenase activities were measured by a functional assay in a longitudinal cohort study . Comparisons were made between human subjects with either: 1) inflammation with a previous history of progressive loss of connective tissue and bone support (n = 14); 2) inflammation and previous history of bone loss but now clinically stable (n = 27); or 3) inflammation and no loss of bone support (n = 17) . Experiments using specific enzyme inhibitors, blocking antibodies and SDS-PAGE fluorograph to identify the pattern of collagen substrate degradation demonstrated that the collagenase activity was derived from neutrophils and not from bacteria or other host cells . Active collagenase activity pooled from 6 sites per subject was respectively 5 and 6-fold higher in the group with progressive loss of connective tissue compared to the groups with either inflamed tissues alone or with inflammation and previous bone loss . In contrast, latent collagenase was increased up to 2 fold higher in the group with inflammation but no bone loss compared to the group with progressive lesions . Moreover, the ratio of active to total collagenase activity was 50% higher in the group with progressive lesions . Although in all subjects successive measurements of site-specific active collagenase 1 month apart demonstrated wide variation (r < 0.50), only in sites with progressive periodontal destruction was there significant increase of active collagenase with time (1.28 x 10(-4) collagenase units per day) . There were also sharp elevations in active enzyme level at the time of detection of loss of connective tissue attachment in specific sites of 8 subjects . At the time of detection of connective tissue attachment loss, there was an overall 40% increase of pooled active collagenase activity in all subjects with progressive loss of connective tissue compared to pre-breakdown sampling times . These data provide strong in vivo evidence for a direct role of active neutrophil collagenase in the pathological destruction of periodontal connective tissue. Clin Diagn Lab Immunol, 1995 Jan, 2(1), 18 - 24 Quantitative reverse transcription-PCR analysis of Legionella pneumophila-induced cytokine mRNA in different macrophage populations by high-performance liquid chromatography; Yamamoto Y et al.; Cytokine production in macrophages infected by bacteria is critical for the course of infection . However, it is not known how infection of macrophages with opportunistic bacteria leads to cytokine production in different populations of cells . Since it is possible that cytokine genes may be differentially regulated by attachment rather than by active infection, the levels of various cytokine mRNAs were measured in alveolar macrophages (AMs), peritoneal resident macrophages (RMs), and peritoneally elicited macrophages (EMs) interacting with Legionella pneumophila by using cytochalasin D-treated macrophages and a newly developed quantitative reverse transcription-PCR procedure with high-performance liquid chromatographic analysis to determine cytokine mRNA formation . Increased levels of interleukin-1 beta (IL-1 beta), IL-6, tumor necrosis factor alpha, granulocyte-macrophage colony-stimulating factor, and macrophage inflammatory protein 2 mRNAs were quantitated in the macrophages responding to L . pneumophila attachment in vitro . Using this technique, we showed that the three different macrophage populations responded differently to bacterial attachment . We found that the levels of IL-6 and granulocyte-macrophage colony-stimulating factor mRNAs induced by the attachment of L . pneumophila to AMs were significantly lower than the levels in RMs but similar to the levels in EMs . Furthermore, the levels of MIP-2 mRNA in the AMs were found to be higher than those in the RMs, but similar levels were found in EMs . IL-1 beta mRNA levels were higher in both AMs and RMs than in EMs, but tumor necrosis factor alpha levels were not different among the three macrophage populations examined . Thus, the responses of macrophages to bacterial attachment in terms of cytokine mRNA levels were readily quantitated by the reverse transcription-PCR assay . However, the results obtained showed different levels of responsiveness of distinct macrophage populations to L . pneumophila attachment, and this could be related to the characteristic nature of the macrophage type examined. J Endod, 1995 Jan, 21(1), 13 - 5 Effect of super-EBA as a root end filling on healing after replantation; Pitt Ford TR et al.; The effect of Super-EBA cement as a root-end filling placed in teeth before replantation was examined in eight molar roots in monkeys . After extraction, root ends were resected, the canals contaminated with oral bacteria, root-end cavities prepared, and fillings of Super-EBA placed before replantation . After 8 wk, the jaws were removed and prepared for histological examination . The tissue response to Super-EBA was very mild, with only a few inflammatory cells being observed at the root end of 3 of the 8 roots filled . Previous work showed a similarly mild response to Intermediate Restorative Material and a very much more severe response to amalgam . It is concluded that the tissue response to Super-EBA as a root-end filling is acceptable and considerably more favorable than that to amalgam. J Math Biol, 1995, 33(4), 388 - 414 Finite time blow-up in some models of chemotaxis; Rascle M et al.; We consider a class of models of chemotactic bacterial populations, introduced by Keller-Segel . For those models, we investigate the possibility of chemotactic collapse, in other words, the possibility that in finite time the population of predators aggregates to form a delta-function . To study this phenomenon, we construct self-similar solutions, which may or may not blow-up (in finite time), depending on the relative strength of three mechanisms in competition: (i) the chemotactic attraction of bacteria towards regions of high concentration in substrate (ii) the rate of consumption of the substrate by the bacteria and (iii) (possibly) the diffusion of bacteria . The solutions we construct are radially symmetric, and therefore have no relation with the classical traveling wave solutions . Our scaling can be justified by a dimensional analysis . We give some evidence of numerical stability. Immunol Invest, 1995 Jan-Feb, 24(1-2), 87 - 93 Leukocyte filtration mechanisms . Factors influencing the removal of infectious agents from red cell concentrates; Steneker I et al.; The purpose of the present overview was to determine the factors influencing the removal of infectious agents from red cell concentrates by filtration . In general, the efficacy of the filtration method depends on the physical as well as the functional properties of blood cells . These properties are highly influenced by the changes exerted on the blood cells during blood collection, processing and storage and the filtration method itself . In particular, the removal of infectious agents of red cell concentrates by filtration will be determined by the type of virus and therewith the binding towards leukocytes, the type of bacteria and holding period before filtration, the deformability of infected cells and the disintegration of cells in the filter. Immunol Invest, 1995 Jan-Feb, 24(1-2), 49 - 71 Reducing the infectivity of blood components--what we have learned; Friedman LI et al.; The safety of the nation's blood supply has improved over the last several years as a result of more intensive donor screening and viral testing . Concurrently, there has been more judicious use of blood components . Although the risk is small, transmission of blood borne viruses, bacteria and parasites can occur . Investigators have studied a myriad of processes for pathogen depletion and/or inactivation, including the use of chemicals, extended storage, filtration, heating, irradiation, photochemicals and washing . Pasteurization, methylene blue and solvent-detergent processes have been introduced in parts of Europe for improving the safety of plasma used for transfusion . The FDA is reviewing a license application for the solvent-detergent process . For red cells, use of highly efficient leukodepletion filters is believed to be equivalent to antibody testing for the prevention of CMV disease transmission . Otherwise, no successful treatments have yet been identified for red cells or platelets . Several photochemicals, which may be useful for treating these components, are being studied . However, there appear to be trade-offs between the extent of pathogen inactivation, platelet or red cell damage, and genotoxicity . These as well as other biological parameters and operational issues will need to be further evaluated before implementation can be considered. Biotechniques, 1995 Jan, 18(1), 142 - 5 Eukaryotic GST fusion vector for the study of protein-protein associations in vivo: application to interaction of ATFa with Jun and Fos; Chatton B et al.; We describe a multipurpose eukaryotic expression vector that incorporates the following features: restriction sites for in-frame insertion of cDNAs of interest between sequences encoding the glutathione-S-transferase (GST) and an oligohistidine element, allowing expression of the corresponding fusion proteins; a phosphorylation site for protein kinase A for in vitro labeling of the fusion protein; a T7 promoter for in vitro transcription and subsequent translation; and signals for single-stranded DNA production in bacteria . We have used this vector to demonstrate the formation in vivo of complexes between the transcription factor ATFa, a member of the family of ATF/CRE binding proteins, and the c-Jun or c-Fos proteins . Such interactions could be detected in crude extracts from cells transfected with vectors expressing the GST-ATFa fusion protein, as well as the c-Jun or c-Fos proteins . Complexes containing both ATFa and either c-Jun or c-Fos were specifically retained on glutathione (GSH)-agarose beads as revealed by immunoblot analyses . We also show that the leucine zipper domain of ATFa is essential for this interaction. Khirurgiia (Mosk), 1995, (4), 43 - 4 {Endotoxemia in crush syndrome}; Kasatkin VN et al.; The crush syndrome developing in casualties in mass catastrophes is the cause of their high mortality rates . We examined 18 children aged from 9 to 14 years with the crush syndrome who suffered during the earthquake in Armenia . Besides other laboratory studies, the test for the titer of antibodies (TA) to endotoxin was performed . Despite the applied complex therapy, the children's condition grew worse on day 8 . The TA level reduced threefold . Six patients received a single infusion of plasma with naturally increased TA titer in a dose of 10 ml/kg . On the following day after TA infusion, the condition of children in the experimental group improved, the clinical picture of endotoxemia in the control group persisted for 10-12 days. Aviakosm Ekolog Med, 1995, 29(1), 4 - 19 {Gravitational mechanisms of interactions of sensory systems in invertebrates in the evolutionary aspect}; Vinnikov IaA; The paper concerns the origin and the interaction of sensory organs in the context of locomotion . The Earth's gravity, light, sound, electrical, mechanical, etc . impacts were the morphogenetic factors of evolution which pushed the gene to elaborate adequate mechanisms for surmounting gravity, i.e . for exercising directed locomotion . Indeed, even some species of bacteria have mobile levers, flagelli . As a rule, the flagellum itself is the carrier of protein sensory molecules perceiving gravity as a mechanic stress, light, chemical ligands, sound, electricity, etc . On the molecular, subcellular, cellular, and organic levels in unicells and most ancient multicellular organisms an attempt has been made to follow the evolution of locomotion substrate and sensory organs and nerve centers interacting with the substrate and each other and, taken together, recognized as the locomotor-sensory system (LMSS). Scand J Infect Dis, 1995, 27(2), 157 - 62 Open lung biopsy provides a higher and more specific diagnostic yield compared to broncho-alveolar lavage in immunocompromised patients . Fungal Study Group; Ellis ME et al.; In order to examine the feasibility and safety of undertaking a larger prospective study to compare the diagnostic yield from concurrent open lung biopsy (OLB) and bronchoalveolar lavage (BAL) in febrile neutropenic patients with pulmonary infiltrates and the impact of such knowledge on clinical outcome, a pilot exploratory study was performed . 13 immunocompromised patients (mainly with haematological malignancy or bone marrow transplantation recipients) were investigated . At least one diagnostic finding in 12 of 13 patients was provided by OLB compared to 4 of 13 patients by BAL . BAL provided 7 specific diagnoses (pneumocystis 1, fungal infection 3, bacterial pneumonia 1, pulmonary haemorrhage 2) whilst OLB provided 12 specific diagnosis (CMV 2, pneumocystis 3, fungal infection 1, bacterial pneumonia 1, pulmonary haemorrhage 4, pulmonary embolism 1) . Five patients with nonspecific interstitial/alveolar inflammation were diagnosed only by OLB . The concordance that the exact same specific diagnoses present in the OLB were found in the BAL was zero . There were 2 minor complications (1 wound infection by OLB, 1 moderate haemorrhage by BAL) . Mortality at 28 days was 8 of 13 patients which in no case was related to either procedure . We suggest that OLB is a safe procedure in such patients, provides superior and more complete diagnostic information compared to BAL and a larger controlled study to investigate the impact of early OLB on the outcome of these patients appears to be justified. Methods Enzymol, 1995, 250, 582 - 614 Mammalian glycosylphosphatidylinositol-anchored proteins and intracellular precursors; Hirose S et al.; Glycosylphosphatidylinositol-anchored proteins can be specifically identified by several methods . PI-PLC digestion analyses, the most widely used technique, can be performed more reliably when conducted with purified protein and phase partitioning to exclude steric effects and when combined with alkaline hydrolysis to control for inositol acylation . Reductive radiomethylation not only can definitively identify a candidate protein as being GPI anchored, but also can provide information on the number of amine components (GlcN, ethanolamine) in the anchor structure . Biosynthetic labeling with anchor precursors is relatively specific when performed with {3H}ethanolamine or {3H}inositol . Incorporation of the precursors additionally can be used to (1) document anchor transfer to primary translation products, (2) identify soluble derivatives of GPI-anchored proteins that have been released from cell surfaces, and (3) localize the site of GPI anchor attachment within a GPI-anchored protein . A pathway for mammalian GP anchor assembly is depicted in Fig . 12 . Initially GlcNAc is transferred to PI . The resulting GlcNAc-PI is then deacetylated to yield GlcN-PI . After that step, several points of divergence are identifiable between the mammalian and T . brucei pathways: (1) all mammalian Man-containing intermediates are built on acylated inositol phospholipids; (2) a proximal phosphoethanolamine is found in mammalian GPI anchor intermediates and is added to Man 1 prior to incorporation of Man 2 and Man 3; (3) no Gal branching substituent is added to the mammalian core glycan; and (4) the most polar mammalian GPI contains a third phosphoethanolamine substituent linked to the 6 position of Man 2. J Mal Vasc, 1995, 20(2), 107 - 12 {Exploration of the various steps of polymorphonuclear neutrophil function in diabetic patients}; Delamaire M et al.; Vascular complications in diabetic patients is a complex, probably multifactorial phenomena involving cellular phagocytosis . The aim of this study was to evaluate polymorphonuclear performance in 61 infection free diabetic patients based on tests of the different cell functions: 1) adhesion:adhesion molecule expression CD11a, CD11b, CD11c; adhesion test on nylon fibers . 2) chemotaxis:chemotaxis under aragose to FMLP (bacteria oligopeptide) and complement fractions . 3) Phagocytosis:latex beads . 4) Bacteriocidal power:chemoluminescence photometric oxidative potential before and after stimulation with opsonized zymosan and PMA; reduction of tetrazolium nitroblue . Results were analyzed according to type of diabetes, glucose control, duration of the disease, history of infection and presence of vascular complications . Results: compared with a group of 30 controls, the diabetic patients had a significant impaired polynuclear chemotaxis function (p < 0.001) with both spontaneous adhesion and increased expression of adhesion molecules (CD 11b, CD 11c) . The chemoluminescence test was increased at the baseline level due to spontaneous polynuclear adhesion and increased production of free radicals . This response decreased after stimulation compared with controls . The type of diabetes, Hb A1c level and history of infection did not appear to have an effect . Inversely, changes in chemotaxis and chemoluminescence were greater in patients with vascular complications . In summary, all the functions of polynuclear neutrophils tested were altered in diabetic patients and could favor vascular complications and infections episodes. Biol Neonate, 1995, 67 Suppl 1, 2 - 17 Potential role of surfactant proteins A and D in innate lung defense against pathogens; van Golde LM; The physiological role of pulmonary surfactant is probably not limited to conferring mechanical stability to the alveoli . Increasing evidence suggests that surfactant components, in particular the hydrophilic surfactant proteins SP-A and SP-D, play potentially important roles in host defense mechanisms . Both SP-A and SP-D are collagenous C-type lectins (collectins) that are structurally and, perhaps, functionally related to collectins in the circulation . As will be discussed in this review, evidence is accumulating that the alveolar collectins SP-A and SP-D could be important components of a first-line defense system against infiltrating pathogenic micro-organisms and viruses. Crit Rev Biotechnol, 1995, 15(2), 105 - 24 Principles and applications of biosensors for bioprocess monitoring and control; Mulchandani A et al.; Biosensors are useful analytical devices that can be integrated with on-line process monitoring schemes . In this article, the principles and applications of these devices for bioprocess monitoring are considered . Several different types of biosensors are described, and the applications and limitations of flow injection analysis (FIA) for these applications are discussed . It is hoped that the background provided here can be useful to researchers in this area. Bull Cancer, 1995, 82(5), 339 - 48 {Identification of human carcinogenic risks in IARC monographs}; Vainio H et al.; For more than twenty years, the IARC has been evaluating the carcinogenic risk to humans of chemicals, groups of chemicals, complex mixtures, occupational exposures, behavioral and life-style exposures, biological agents, such as bacteria and viruses, and physical agents, such as radiation, on the basis of published studies of carcinogenicity in humans and laboratory animals . This paper includes the list established by IARC of substances carcinogenic to humans. Probl Tuberk, 1995, (3), 37 - 41 {Use of bronchovaxom in the treatment of patients with chronic obstructive bronchitis and bronchial asthma}; Shmelev EI et al.; Polyvalent vaccine bronchovaxom (BV) was studied for efficacy against chronic obstructive bronchitis and bronchial asthma exacerbation . Good results were reported: reduction of disability duration, number of recurrences, cough intensity, discharged sputum . Laboratory tests discovered that BV brought about IgE decrease, IgA, T3, T4/T8 increase in bronchoalveolar lavage. Neoplasma, 1995, 42(1), 25 - 30 Combined radioprotective effect of Broncho-Vaxom and WR-2721 on hemopoiesis and circulating blood cells; Mackova NO et al.; Possibilities of combined radioprotection, using preirradiation WR-2721 administration and post- or preirradiation Broncho-Vaxom administration in lethally whole-body gamma-irradiated mice were investigated . Combined modality treatments were more effective than individual treatments alone . The ameliorative influence of the combined radioprotection could be explained by the hemopoietic stem cell protection with WR-2721 and the stimulation by Broncho-Vaxom. Annu Rev Immunol, 1995, 13, 251 - 76 Interleukin-12: a proinflammatory cytokine with immunoregulatory functions that bridge innate resistance and antigen-specific adaptive immunity; Trinchieri G; Interleukin-12 (IL-12) is a heterodimeric cytokine produced mostly by phagocytic cells in response to bacteria, bacterial products, and intracellular parasites, and to some degree by B lymphocytes . IL-12 induces cytokine production, primarily of IFN-gamma, from NK and T cells, acts as a growth factor for activated NK and T cells, enhances the cytotoxic activity of NK cells, and favors cytotoxic T lymphocyte generation . In vivo IL-12 acts primarily at three stages during the innate resistance/adaptive immune response to infection: 1 . Early in the infection, IL-12 is produced and induces production from NK and T cells of IFN-gamma, which contributes to phagocytic cell activation and inflammation; 2 . IL-12 and IL-12-induced IFN-gamma favor Th1 cell differentiation by priming CD4+ T cells for high IFN-gamma production; and 3 . IL-12 contributes to optimal IFN-gamma production and to proliferation of differentiated Th1 cells in response to antigen . The early preference expressed in the immune response depends on the balance between IL-12, which favors Th1 responses, and IL-4, which favors Th2 responses . Thus, IL-12 represents a functional bridge between the early nonspecific innate resistance and the subsequent antigen-specific adaptive immunity. Reprod Nutr Dev, 1995, 35(3), 249 - 66 Digestion of a dehydrated lucerne/barley diet (70:30) in defaunated, Isotricha-monoinoculated and mixed-fauna-inoculated rumen in sheep; Senaud J et al.; Two adult Texel sheep fitted with permanent rumen fistulae were defaunated for 12 weeks in a preliminary experiment . They were then reinoculated with Isotricha sp for 11 weeks, and finally with a mixed fauna (Entodinium, Epidinium, Eudiplodinium and Isotricha) for 10 weeks . They were fed a diet composed of dehydrated lucerne (700 g/d), pelleted barley grain (300 g/d), grass hay (100 g/d) and wheat straw (50 g/d) in one daily meal . Isotricha and the ciliates of the mixed fauna developed rapidly, reaching a maximum concentration 9-17 d after inoculation . Their concentration then fell for 2-3 d and finally stabilized at values close to 10(4)/ml for Isotricha, Eudiplodinium and Epidinium and 2.5 x 10(5)/ml for Entodinium . The estimated ciliate biomass in the rumen was 1.8 g l-1 for a volume of 10.2 ml l-1 in the Isotricha-monoinoculated sheep and 4.7 g l-1 for a volume of 25.0 ml l-1 in mixed-fauna-inoculated sheep . The concentration of Isotricha was unaffected by the inoculation of mixed fauna during the third part of the experiment . No difference in total rumen bacteria counts was observed between defaunated, monoinoculated and completely refaunated animals . The total adenylic nucleotide concentration in filtered rumen juice was 4 times higher in faunated sheep just before feeding and 4-9 times lower after feed intake as compared to defaunated animals; the energy charge was always higher in faunated animals . These results are discussed in relation to the digestive activity and biomass of the protozoa . The in situ degradation of lucerne stems was the highest in the mixed-faunated sheep over a retention time interval of 6-18 h . The values obtained in Isotricha-monoinoculated sheep were intermediate between the defaunated and the mixed-faunated states . The digestibility in the whole digestive tract of dietary dry matter, organic matter, nitrogen and neutral detergent fibre was unaffected by the addition of either Isotricha or the mixed fauna in defaunated rumens . The total volatile fatty acid concentration was higher (p < 0.05) in the defaunated sheep between 6 and 18 h after feeding . The molar proportion of butyrate was significantly (p < 0.05) increased by the presence of mixed fauna in the rumen while that of propionate and valerate was lower . A non-significant decrease in acetate was also observed . The concentration of NH3-N in the rumen was significantly increased (p < 0.05) by the presence of the mixed fauna in the rumen but was unaffected or slightly reduced by Isotricha alone.(ABSTRACT TRUNCATED AT 400 WORDS) Can J Microbiol, 1995, 41 Suppl 1, 50 - 4 Low molecular weight complexed poly(3-hydroxybutyrate): a dynamic and versatile molecule in vivo; Reusch RN; It is increasingly clear that poly(3-hydroxybutyrate) (PHB) is not just an inert storage polymer, confined to certain bacteria, but a ubiquitous, interactive, solvating biopolymer involved in important physiological functions . Low molecular weight PHB, complexed to other macromolecules (c-PHB), is widely distributed in biological cells, being found in representative organisms of nearly all phyla . Complexation modifies the physical and chemical properties of c-PHB, allowing it to pervade aqueous as well as hydrophobic regions of the cell, and as a result c-PHB can be found in cytoplasm and intracellular fluids as well as in membranes and lipoproteins . The lipidic homopolymer associates with other macromolecules primarily via its ester carbonyl oxygens, which can act as hydrogen-bond acceptors or as ligands for coordinate bonds to cations . The spacing of the electron-donating groups along the flexible backbone allows for multiple bonding interactions, and forms the basis for the ability of c-PHB to bind to proteins, or to form ion-conducting complexes with salts . The singular ability of c-PHB to dissolve salts and facilitate their transfer across hydrophobic barriers defines a potential physiological niche for c-PHB in cell metabolism. J Pediatr Endocrinol Metab, 1995 Jan-Mar, 8(1), 3 - 10 Exposure to superantigens as an immunogenetic explanation of type I diabetes mini-epidemics; Trucco M et al.; Insulin-dependent diabetes mellitus (IDDM) is considered to be an autoimmune disease . Autoimmune diseases result from the breakdown of self-tolerance elicited by environmental factors on a susceptible genetic background . This reduced tolerance allows immunocompetent cells to attack normal structures present on an individual's tissues . The study of the T cells present in the islets of patients who died at the onset of the disease allowed the recognition of "superantigens" as etiopathogenetic factors in the development of IDDM . Since superantigens are the product of bacteria or viruses able to quickly stimulate a large number of the peripheral T cells sharing the same T cell receptor V beta segments once presented by certain HLA class II molecules, a number of observations in genetics, immunology, virology and epidemiology can now find a unifying explanation. Braz J Med Biol Res, 1995 Jan, 28(1), 61 - 4 Health problems of Callithrix jacchus in captivity; Diniz LS et al.; A 20-year retrospective study was carried out to determine the prevalence of diseases occurring in 265 (155 males and 110 females) common marmosets (Callithrix jacchus), kept at the Sao Paulo Zoo . A total of 564 clinical disorders were detected: digestive problems, 50.2% (protozoa, 44.3%; helminths, 26.3%; bacteria, 7.0%; non-especific causes, 22.4%); respiratory problems (pneumonia), 16.3%; injuries, 13.8%; nutritional deficiency, 4.4%; reproductive problems (obstetrical) 2.5%; septicemia, 1.6%; circulatory problems, 1.6%; sporadic cases, 4.0%; inconclusive cases, 5.5% . Little attention has been paid to the sanitary care of marmosets in Brazil . Since most of the available information comes from the international literature, the occurrence of major diseases and their etiological agents in Brazil are relevant data. Crit Rev Toxicol, 1995, 25(4), 281 - 346 The influence of salinity on the toxicity of various classes of chemicals to aquatic biota; Hall LW Jr et al.; The objective of this study was to review all available aquatic toxicity literature regarding the effects of salinity on the toxicity of various classes of inorganic and organic chemicals . Toxicity data for studies in which toxicity was assessed at various salinities were organized by chemical classes and trophic groups . Seventy percent of the studies were conducted with either crustaceans or fish . The other 30% were with mollusks, annelids, zooplankton, bacteria, phytoplankton, or fungi . Results from 173 data entries showed that negative correlations (toxicity increasing with decreasing salinity) were reported most frequently (55%), followed by no correlations (27%) and positive correlations (18%) . The toxicity of most metals such as cadmium, chromium, copper, mercury, nickel, and zinc was reported to increase with decreasing salinity . This finding is likely related to the greater bioavailability of the free metal ion (toxic form) at lower salinity conditions . There was generally no consistent trend for the toxicity of most organic chemicals with salinity . The one exception to this was reported with organophosphate insecticides, the toxicity of which appeared to increase with increasing salinity . Physiological characteristics of the various test species were important in determining the toxicity of the various classes of chemicals at a range of salinities . Results from various studies showed that euryhaline species were more resistant to toxic conditions at isosmotic salinities due to minimization of osmotic stress . Specific examples showed that fish were more resistant to toxic chemicals at middle salinities when compared with either lower or higher extremes . Life history and ecology of test species were important factors to consider when interpreting salinity/contaminant interaction data. Ann Chir, 1995, 49(5), 423 - 6 {Effects of intraperitoneal insufflation on hematogenous seeding of abdominal infections . Preliminary results of an experimental study in rats}; Dugue L et al.; Most laparoscopic procedures require the creation of a pneumoperitoneum . In order to evaluate the potential hazards of bacteriemia related to insufflation, we conducted a study in the rat . Two groups of 20 Wistar rats were used for this study . Peritonitis was induced by opening the terminal ileum . Twenty-four hours later, 20 rats were insufflated at a mean pressure of 6 mm Hg (Group I) . After one hour of insufflation, an hemoculture was performed via direct intracardiac puncture and in the other group of 20 non-insufflated rats (Group NI) . Five of the 18 hemoculture were positive in the Gr . I (27.7%) and 6 out of 20 in the Gr . NI (30%) (chi 2 = 0.238 p = 0.62 non significant difference) . These results suggest that insufflation does not facilitate hematogenous dissemination of bacteria from intraperitoneal sepsis in this animal model. Cell Mol Biol Res, 1995, 41(1), 49 - 58 The tyrosine phosphorylation of a p72syk-like protein in activated murine resident peritoneal macrophages; Abu-Lawi KI et al.; As a marker of macrophage activation, IL-1 alpha was measured after stimulation of murine resident peritoneal macrophages (RPM) with endotoxin-associated protein (EP) . Significant IL-1 alpha was produced by EP-stimulated RPM from both C3H/OuJ and C3H/HeJ mouse strains . This EP-mediated IL-1 alpha production was blocked by tyrosine kinase inhibitors including genistein and tyrphostin, suggesting the involvement of a protein tyrosine kinase in the activation of RPM by EP . Immunoblot analysis using antiphosphotyrosine antibody showed that EP induces the tyrosine phosphorylation of a 71 kD protein (p71) . The p71 and the spleen tyrosine kinase p72syk found in other cell types share common features including: similar molecular weight, PKC independent tyrosine phosphorylation, and inhibition of phosphorylation by piceatannol . Furthermore, immunoblot analysis using anti-p72syk antibody detected the p72syk kinase in EP-activated RPM . These results suggest that the activation of RPM involves an early tyrosine phosphorylation of p72syk or a p72syk-like protein. Implant Dent, 1995 Spring, 4(1), 21 - 6 The use of high-density polytetrafluoroethylene membrane to treat osseous defects: clinical reports; Bartee BK; Alveolar bone resorption can result from tooth loss, periodontal disease, or trauma . Guided tissue regeneration is used in an attempt to exclude tissues devoid of osteogenic potential from a bone defect or cavity and promote new bone growth to replace missing osseous structure . Many types of barrier membranes have been used, but none have been found to be ideal for every clinical situation . Macroporous membranes, such as expanded polytetrafluoroethylene, require primary closure and a second surgical procedure for their removal . Macroporous membranes can incorporate bacteria and may become infected if exposed in the oral cavity . Membranes manufactured of resorbable polymers require primary closure of the augmentation site and exhibit variable patterns of resorption, introducing a degree of unpredictability into the procedure . The use of high-density polytetrafluoroethylene membrane to promote deposition of bone for ridge augmentation in the oral cavity is described . Two clinical reports are presented. Eur J Histochem, 1995, 39(2), 157 - 64 Acid phosphatase activity in mating type I and mating type II cell lines of Paramecium primaurelia; Ramoino P et al.; The cellular acid phosphatase content, a marker enzyme for lysosomal activity, in Paramecium primaurelia mating type I and mating type II cells was determined by optical laser scanning microscopy . The naphthol AS-TR phosphatase-hexazotized pararosaniline method was used to visualize acid phosphatase activity by the light microscopy . Cell lines of both mating types were tested during culture life, from the early log phase to the death phase . The amount of acid phosphatase was higher in mating type II than in mating type I until the onset of the stationary phase, and then the values reversed . Indeed, during the log phase of growth, mating type II cells formed a higher number of food vacuoles, so that, by taking up a higher amount of bacteria, they sooner became deprived of food . It is suggested that, by lacking nutrients, their synthesis activities and acid phosphatase content were reduced as compared with mating type I cells. Arch Insect Biochem Physiol, 1995, 29(3), 269 - 80 Biochemical characterization, developmental expression, and induction of the immune protein scolexin from Manduca sexta; Kyriakides TR et al.; The immune protein, scolexin, a bacteria-induced, larva-specific protein from Manduca sexta, was shown to exist in the hemolymph in two isoelectric forms designated herein as scolexin-1 and scolexin-2 (native M(r) approximately 72 kd) . These two charge isomers appeared to share the same amino acid composition . Scolexin is composed of two subunits (peptide M(r) approximately 36 kd) that possess the same N-terminus . Scolexin-2 was subjected to glycosyl composition analysis, revealing the presence of galactose, glucose, mannose, xylose, and sialic acid residues . Hybridization of epidermal RNA with oligonucleotides deduced from the scolexin N-terminal sequence showed a continuous decline in mRNA following day 0 of the 5th larval instar . By employing in vitro protein labelling, it was found that organ cultures of the epidermis from immune larvae showed a greater ability over that of naive epidermal cultures to synthesize scolexin; these data reflected the inducible response seen in the hemolymph, and confirm other data indicating that the epidermis is an important site of scolexin biosynthesis. Cell Motil Cytoskeleton, 1995, 30(3), 171 - 82 Monoclonal anti-dipeptide antibodies cross-react with detyrosinated and glutamylated forms of tubulins; Kuriyama R et al.; Two monoclonal antibodies, GLU-1 and A1.6, raised against gamma-L-glutamyl-L-glutamic acid dipeptide (Glu-Glu) and Ca(2+)-dependent ATPase from Paramecium, respectively, recognized the dipeptide Glu-Glu sequence . Whereas the antibodies immunofluorescently stained very few, if any, cytoskeletal fibers in cultured mammalian cells, almost all interphase as well as mitotic spindle microtubules became visible after treatment of cells with carboxypeptidase A . Immunoblot analysis demonstrated intense cross-reaction of the antibodies to the alpha-tubulin subunit . alpha-Tubulin isotypes produced as fusion proteins in bacteria were labeled by both the antibodies only when the proteins did not contain a tyrosine residue at the C terminus, indicating that GLU-1 and A1.6 specifically recognize the detyrosinated form of alpha-tubulin . When microtubule protein purified from brain was probed, not only alpha-but also, to a lesser extent, beta-tubulin were revealed by the dipeptide antibodies . A synthetic tripeptide YED containing one glutamyl group linked to the second residue of the peptide via the gamma position was also recognized by the antibodies . Since this peptide sequence corresponds to the amino acid sequence of polyglutamyated class III beta isotype at amino acid position 437 to 439, it is suggested that GLU-1 and A1.6 are able to recognize the glutamylated form of beta-tubulin . These results indicate that the C-terminal Glu-Glu sequence displays strong antigenicity, and the antibodies recognize the sequence present in the C terminus of the detyrosinated form of alpha-tubulin and the glutamyl side chain of beta-tubulin . Particularly strong immunoreaction was detected with ciliary and flagellar microtubules; thus, stable axonemal microtubules appear to be rich in post-translationally modified tubulin subunits. Nippon Rinsho, 1995 Jan, 53(1), 250 - 7 {Water channel family proteins}; Ishibashi K et al.; 17 members of MIP family from bacteria, yeast, plants and animals are compared in this review . These proteins appear to function in (1) water channels (CHIP, WCH-CD, MIWC, AQP3, gTIP, RD28, TobRB7), (2) neurogenesis (Bib), (3) small-molecule-permeating channels (MIP, AQP3, NOD, Glpf), (4) unknown function (WCH-3, AtRB7, Pea R7A, FPS1) . However, the biological functions are not well established . The most conserved residues in the first and the second halves of all MIP family proteins are asparagine-proline-alanine (NPA) sequences in the loops (NPA boxes) . This structural similarity may lead to functional similarity (water and/or small molecule permeation) . This signature sequence for the MIP family will facilitate the identification of new protein members of this family. Mutat Res, 1995 Jan, 341(3), 141 - 9 The genetic toxicology of ethylenethiourea: a case study concerning the evaluation of a chemical's genotoxic potential; Elia MC et al.; Ethylenethiourea (ETU) is a metabolite, environmental degradation product and minor technical impurity of the ethylenebisdithiocarbamate (EBDC) class of fungicides . The genetic toxicology of ETU is important given that ETU causes thyroid tumors in rodents and liver tumors in mice . Although it is clear that ETU induces thyroid tumors via a non-genotoxic, threshold mechanism, the role ETU plays in inducing mouse liver tumors remains to be fully elucidated . Recently, Dearfield (Mutation Res., 317, 111-132, 1994) reviewed the genetic toxicology of ETU, and concluded that, although ETU is not a potent genotoxic agent, it is weakly genotoxic . This view stands in contrast to reports from several independent authorities that have generally concurred that ETU is not a mammalian genotoxin (IARC, 1987; MAFF, 1990; NTP, 1992; FAO/WHO, 1994) . These conflicting reports highlight a generic problem in genotoxicity safety assessment: although individual test results typically yield either a positive or negative response, the overall evaluation of an extensive battery of tests for a particular chemical rarely yields an unambiguous conclusion . Recently, Mendelsohn et al . (Mutation Res., 266, 43-60, 1992) showed that the response of a chemical to a battery of genotoxicity tests is not a dichotomous (i.e., either positive or negative) property, but rather, appears to be a continuous property that ranges from strongly negative to strongly positive . We have used these data, together with a four-step weight of the evidence procedure, to evaluate ETU . Our analysis indicates that ETU is not genotoxic in mammalian systems and suggests that ETU likely induces mouse liver tumors by a non-genotoxic mechanism. Fertil Steril, 1995 Jan, 63(1), 158 - 65 Carbohydrate moieties on sperm surface: physiological relevance; Kurpisz M et al.; OBJECTIVES: To study the cross-reactions between mouse monoclonal antisperm antibodies and somatic cells or bacteria, to identify the antigenic determinants responsible for such cross-reactions, and to correlate between the antibody function and determinant recognition . DESIGN: Activities of monoclonal antibodies (mAbs) were characterized by immunosorbent assay of RIA technique; sperm epitopic characterization was performed in lectin-blocking and sugar competitive assays and correlated with functional assays . SETTING: Procedures were performed in a university laboratory . RESULTS: The extensive cross-reactivity between antigenic determinants of sperm, erythrocytes, and bacteria (but not bacterial deglycosylated lipopolysaccharides) was observed . The analytic procedures indicated predominant mAb reactions to carbohydrates such as fucose, galactose, mannose, N-acetylglucosamine and N-acetylgalactosamine . Approximately half of the 30 tested mAbs interfered in the functional assays, that is, sperm agglutination, immobilization, and zona-free penetration . CONCLUSIONS: Sperm carbohydrates seem to induce antibody reactions to common antigenic determinant(s) present on gametes, somatic cells, and infectious agents . Thus molecular mimicry between bacteria and sperm can be a major factor inducing antisperm immunological reactions . Obtained antisperm mAbs, reacting to glycosylated epitopes, presented very strong properties in sperm agglutination and/or immobilization . This did not correlate with inhibiting properties of some antibodies in xenogeneic zona-free penetration test, that is, this assay possibly is not based on oligosaccharide mediation. Crit Rev Clin Lab Sci, 1995, 32(3), 265 - 98 Physiology and pathophysiology of complement: progress and trends; Morgan BP; The complement system comprises a family of at least 20 plasma and membrane proteins that interact in a tightly regulated cascade system to destroy invading bacteria and prevent the deposition of immune complexes in the tissues . This brief review addresses the basic mechanisms of complement activation and control and describes the active fragments produced during complement activation . The biological importance of the complement system is amply illustrated in patients with complement deficiencies, who are susceptible to bacterial infections and immune complex diseases . The involvement of complement in other immunological diseases is an expanding area of clinical research, supported by the development of new assays for the identification of complement activation . This area is discussed here with particular reference to neurological diseases . A promising new prospect involves the use of complement inhibitory molecules in therapy of complement-mediated disease and this exciting area is also discussed . Novel physiological roles of complement also are being revealed and new evidence that complement and complement receptors play an important role in reproduction is summarized . It is hoped that this brief overview will convey some of the enthusiasm currently pervading research in this underappreciated area of immunology. Microbiol Immunol, 1995, 39(8), 623 - 7 Functional activities of antibodies directed against surface lipoproteins of Borrelia hermsii; Sambri V et al.; Enriched preparations for mouse polyclonal immunoglobulin G (IgG) antibodies reactive with surface-exposed epitopes (Ab-SEE) of the 22-kDa and 24-kDa membrane lipoproteins of living Borrelia hermsii (HS 1 strain) cells were obtained by an antibody absorption technique using living spirochetes . In vitro, the antibody preparations both inhibited spirochetal growth and were borreliacidal in the presence of complement . The monovalent Fab antibody fragments, prepared from antibody-enriched preparations, did not inhibit the growth of the bacteria, whereas they killed the bacteria in the presence of complement . The two-dimension gel electrophoresis of B . hermsii cells showed that 3H-labeled fatty acids incorporated into the 22-kDa and 24-kDa lipoproteins were resolved into one and three compact spots, respectively . The spots were recognized by the Ab-SEE preparations reactive with the 22-kDa and 24-kDa proteins, by Western blotting. Verh K Acad Geneeskd Belg, 1995, 57(3), 229 - 47; discussion 247-8 Immunotherapy with bispecific antibodies; Thielemans KM; With rare exceptions, monoclonal antibodies as therapeutic agents for cancer, have had limited anti-tumor effects . Attempts to improve the cytotoxicity of antibodies by using them as guiding agents for immune effector cells, have been developed . Numerous in vitro studies have shown that T lymphocytes can be targeted towards any target cell by using bispecific antibodies with specificity of the CD3/TCR complex and a target cell antigen . This approach works extremely efficiently in vitro and justified the hope that bispecific antibodies might be useful for focusing T cells towards tumor cells in vivo . Therefore we have produced bispecific antibodies directed against the membrane expressed idiotype of the murine B cell lymphomas BCL1 and 38C13, and against the murine CD3 complex present on all T lymphocytes . The dual specificity of the hybrid-hybridoma produced bispecific antibodies could be demonstrated by flow cytometry, the induction of T cell proliferation, the induction of IL-2 secretion by polyclonal T cells, and redirected lysis of the relevant target cells . Immunotherapy of tumor bearing animals demonstrated that bispecific antibodies could efficiently target T cells towards the tumor cells, that tumor cell--T cell bridging is established in vivo, and that both T cell subsets contribute to tumor regression resulting in long-term survival and cure of the lymphomas . To streamline the production of bispecific antibodies we recently engineered recombinant bispecific proteins produced by bacteria . Two scFv fragments, one derived from the anti-tumor antibody and one derived from the anti-mouse CD3 monoclonal antibody were produced in E . Coli . Both antibody fragments could be easily purified by IMAC chromatography . After in vitro refolding of the recombinant proteins the antigen specificity was retained . Both scFv were expressed as one fusion protein by engineering a 15 amino acid linker between the antibody fragments . The purified bispecific fusion protein behaved in vitro tests as active as the hybrid-hybridoma produced bispecific antibody (induction of T cell proliferation and lymphokine secretion, retargeted lysis of tumor cells) . Also the fusion protein is able to target T cells towards the tumor cells in vivo, resulting in long term survival of the treated animals . In conclusion, these studies form a sound basis for further investigation of the potential of bispecific antibodies for treatment of patients with cancer. Rocz Panstw Zakl Hig, 1995, 46(1), 81 - 9 A statistical evaluation of the occurrence of keratinolytic fungi in the sediments of two dam reservoirs; Ulfig K; Sediments from two dam reservoirs "Przeczyce" and "Plawniowice" were examined for keratinolytic fungi . The results show the dependence of keratinolytic fungi in sediments on the degree of water contamination with sewage . Ch . keratinophilum is species associated with sewage inputs to superficial water. Gene, 1994 Dec 30, 151(1-2), 325 - 8 Construction of rare restriction site (NotI, SacII and ClaI) linking libraries and sequence-tagged site analysis of single-copy clones from a human chromosome-3-specific library; Atchison L et al.; We have constructed rare restriction-site (NotI, SacII and ClaI) chromosome 3 (Chr 3)-specific linking libraries in a plasmid-based vector by mass transfer of a lambda phage human Chr-3-specific library (LA03NS01-ATCC57717) into pUC18 . Total plasmid DNA isolated from the plasmid-based Chr-3-specific library was digested with either ClaI, NotI or SacII . Linear molecules were separated from undigested circles by pulsed-field polyacrylamide-gel electrophoresis . Purified linear molecules were circularized with T4 DNA ligase and transformed into bacteria . The resulting clones were greatly enriched for sequences recognized by the original restriction endonuclease used for digestion (83 to 95%) . These sublibraries are composed of 600 (NotI) 1000 (SacII) or 30,000 (ClaI) clones . Thus, this procedure allows for easy isolation of Chr-3-specific DNA clones containing a variety of rare restriction sites . Sequence-tagged site (STS) data are also presented for five site-specifically mapped Chr-3-specific DNA clones . These studies may facilitate the construction of region specific linking libraries for mapping of various disease-specific loci on Chr 3. Gene, 1994 Dec 30, 151(1-2), 23 - 8 Gene organization in the dnaA-gyrA region of the Streptomyces coelicolor chromosome; Calcutt MJ; The gene organization has been determined for an 8-kb portion of the Streptomyces coelicolor chromosome close to the origin of DNA replication (oriC) . Hybridization and DNA sequence analyses revealed the presence of five open reading frames (ORFs) oriented in the same direction as the proximal dnaA and dnaN genes . The deduced products of three of the ORFs have been identified as the S . coelicolor homologs of RecF, GyrB and GyrA . These genes are usually clustered in the dnaA region of bacterial chromosomes . In S . coelicolor however, the usual gene arrangement is altered . The recF gene is flanked by two ORFs, one of which encodes a protein with significant similarity to 6-phosphogluconate dehydrogenases (6PGDH), an enzyme that is not immediately linked to DNA metabolism. J Biol Chem, 1994 Dec 30, 269(52), 33159 - 63 Insect immunity . Septic injury of Drosophila induces the synthesis of a potent antifungal peptide with sequence homology to plant antifungal peptides; Fehlbaum P et al.; In response to a septic injury (pricking with a bacteria-soaked needle) larvae and adults of Drosophila produce considerable amounts of a 44-residue peptide containing 8 cysteines engaged in intramolecular disulfide bridges . The peptide is synthesized in the fat body, a functional homologue of the mammalian liver, and secreted into the blood of the insect . It exhibits potent antifungal activity but is inactive against bacteria . This novel inducible peptide, which we propose to name drosomycin, shows a significant homology with a family of 5-kDa cysteine-rich plant antifungal peptides recently isolated from seeds of Brassicaceae . This finding underlines that plants and insects can rely on similar molecules in their innate host defense. J Biol Chem, 1994 Dec 30, 269(52), 32932 - 6 Mechanisms of the transfer of aminoacyl-tRNA from aminoacyl-tRNA synthetase to the elongation factor 1 alpha; Reed VS et al.; Aspartylation of mammalian tRNAAsp by bacteria-expressed human aspartyl-tRNA synthetase (hDRS) was examined . The kinetics of the aspartylation of tRNA was consistent with the following reaction pathway, {formula: see text} where E, represents aspartyl-tRNA synthetase . A set of rate constants was obtained which fit single turnover time courses at varying concentrations of the enzyme, tRNA, and AMP using the SAAM program . The dissociation of Asp-tRNA (k3) was found to be rate limiting . The elongation factor 1 alpha (EF1 alpha) and GTP stimulated the hDRS aspartylation . The stimulation depended on the presence of both EF1 alpha and GTP . Kinetic analysis indicated that EF1 alpha formed a complex with the hDRS-Asp-tRNA complex and stimulated the dissociation of Asp-tRNA . In the presence of 0.5 M NH4Cl, which enhances the binding of Asp-tRNA by EF1 alpha, hDRS-bound Asp-tRNA can be transferred directly to EF1 alpha . The implications of these results on the function of the multi-tRNA synthetase complex will be discussed. Proc Natl Acad Sci U S A, 1994 Dec 20, 91(26), 12818 - 22 Dynamic interaction between Isp45 and mitochondrial hsp70 in the protein import system of the yeast mitochondrial inner membrane; Kronidou NG et al.; The protein import system of the yeast mitochondrial inner membrane includes at least three membrane proteins that presumably form a transmembrane channel as well as several chaperone proteins that mediate the import and refolding of precursor proteins . We show that one of the membrane proteins, Isp45, spans the mitochondrial inner membrane yet is extracted from this membrane at high pH . Solubilization of mitochondria with a nonionic detergent releases Isp45 as a complex with the chaperones mitochondrial hsp70 (mhsp70) and GrpEp . Both chaperones reversibly dissociate from Isp45 upon addition of ATP or adenosine 5'-{gamma-thio}triphosphate, suggesting that dissociation requires the binding of ATP . Control experiments indicate that the interaction between mhsp70 and Isp45 occurs in the intact mitochondria . We propose that Isp45 lines the inside of a proteinaceous channel across the inner membrane and that it is the membrane anchor for an ATP-driven "import motor" composed of mhsp70 and GrpEp . This arrangement is reminiscent of the protein transport systems of the yeast endoplasmic reticulum and the bacterial plasma membrane. Proc Natl Acad Sci U S A, 1994 Dec 20, 91(26), 12594 - 8 The outer surface protein A of the spirochete Borrelia burgdorferi is a plasmin(ogen) receptor; Fuchs H et al.; The spirochete Borrelia burgdorferi is the causative agent of Lyme borreliosis (Lyme disease) and is transmitted to mammalian hosts by tick vectors . In humans, the bacteria induce a complex disease, which involves the skin, joints, heart, and nervous system . However, the pathogenic principles of this multisystem illness are far from being understood . To disseminate from the site of the tick bite and invade multiple organ sites, spirochetes have to penetrate normal tissue barriers, such as vascular basement membranes and other organized extracellular matrices . Substantial evidence from other invasive bacterial infections suggest that spirochetes may use endogenous or host-derived enzymes--in particular, proteinases--for this purpose . Here we show that B . burgdorferi binds human plasmin(ogen)--mainly via its outer cell surface lipoprotein A . Binding of plasminogen to spirochetal receptor leads to an accelerated formation of active plasmin in the presence of host-derived plasminogen activator . The cell-surface-associated plasmin cannot be regulated by the serum inhibitor alpha 2-antiplasmin and degrades high-molecular-weight glycoproteins, such as fibronectin . It is suggested that the acquisition of host-derived proteinase plasmin(ogen) contributes to the pathogenicity of B . burgdorferi. J Biol Chem, 1994 Dec 16, 269(50), 31770 - 6 An adherens junction protein is a member of the family of lactose-binding lectins; Chiu ML et al.; We previously described a pig junction protein of M(r) 37,000 found in oral epithelium but not in epidermis, limited to suprabasal cells, and colocalizing by immunofluorescence with adherens junction proteins . A 1.1-kilobase pair cDNA of the 37-kDa protein yielded an open reading frame encoding a 323-amino acid protein of 35,852 Da, and Northern analysis demonstrated a band of 1.2 kilobases in tongue RNA . Secondary structure predictions indicate that the 37% identical 16-17-kDa N- and C-terminal domains from beta-sheet-rich barrels linked by a compact proline-rich segment . The protein is 72% identical in amino acid sequence and shares symmetrical two-domain structure with L-36, a lectin of unknown function from rat intestine, indicating that the 37-kDa protein is the porcine form of L-36 . Of the homologous lactose binding lectins known, two others, invertebrate lectins, share this symmetrical structure . Expression of the C-terminal domain of the pig lectin in bacteria yields a lectin which binds lactosyl-Sepharose, and binding is inhibited by lactose . The expressed protein binds a glycoprotein of 120 kDa from pig tongue epithelium on Western blots, and this is also inhibited by lactose . The findings suggest that the lectin function may be involved in the assembly of adherens junctions. Cancer Res, 1994 Dec 15, 54(24), 6297 - 301 Characterization and localization of the TCL-1 oncogene product; Fu TB et al.; The TCL-1 gene maps at chromosome 14q32.1 and is activated in T cell leukemias and lymphomas by either chromosome translocations or inversions that juxtapose the TCL-1 gene to the alpha/delta or the beta locus of the T cell receptor . The open reading frame of the TCL-1 gene, coding for a protein of 114 amino acids, was expressed in bacteria and antisera were raised against it . The antibodies recognized the predicted TCL-1 M(r) 14,000 protein product in cells expressing TCL-1 mRNA . Cell fractionation experiments indicated that the TCL-1 protein is present in the microsomal fraction . These results were confirmed by confocal microscopy . The TCL-1 protein has considerable sequence similarities to the product of the MTCP-1 gene on chromosome Xq28, which is involved in T cell lympho-proliferative diseases . Thus, TCL-1 may represent a member of a novel family of genes involved in lymphoid proliferation and/or survival and in T cell malignancies. Orv Hetil, 1994 Dec 11, 135(50), 2749 - 54 {Skin as an immune system}; Hunyadi J et al.; Skin consists of three structurally and functionally distinct compartments containing resident and nonresident cells . These cells cooperative with humoral pathway of immune system in defence of healthy skin . Resident and nonresident cells are able to initiate inflammatory or immune processes of skin, although interstitial reactions (activation by bacteria, immunoglobulin or complement) also take place in such processes . In normal conditions nonresident cells can migrate through vascular endothel, however, PMN granulocytes and B lymphocytes cannot . Resident and nonresident cells of skin are capable of exerting a wide range of immunomodulatory effects, among them keratinocytes are of distinguished significance producing arachidon metabolites and IL-1 as well . Activated skin cells might induce chemotactic migration of distinct white blood cells normally absent or only a few cells being present in healthy skin . Cell migration into the interstitial space of skin is mediated by adhesion molecules expressed on cell surface of migrating cells, vascular endothel cells and on keratinocytes . Composition and density of adhesion molecules vary by type of stimuli, therefore various cytokines might induce distinct reactions mediated by certain population of cells . In addition, initiation and progress of immune- or inflammatory reactions are determined by the involved cell-populations as well . Normal regulation provides appropriate control and termination of the reaction, however, uncontrolled regulation results in development of pathological state. Ned Tijdschr Geneeskd, 1994 Dec 10, 138(50), 2496 - 500 {Risk of tuberculosis in the inadequate handling of refugees seeking asylum}; van Loenhout-Rooyackers JH; OBJECTIVE . To describe screening for tuberculosis in a temporary centre for refugees, and contact investigation after a patient with open tuberculosis had been identified . DESIGN . Descriptive . SETTING . Regional Communal Health Centre Nijmegen . METHOD . The refugees waiting for official status were housed on three cruise ships during the winter period November 1993 to April 1994 . Ventilatory capacity was insufficient . The people had thorax radiograms after 10 days on average as a screening for tuberculosis . When abnormalities were found sputum was examined (Ziehl-Neelsen staining and culture) . The workers of the centre were Mantoux-tested . RESULTS . Of the 834 persons screened 4 had open tuberculosis . Contact investigation yielded three cases of infection among 5 contacts in the first ring, and 11 cases of infection and 2 cases of tuberculosis (one infiltrative pulmonary tuberculosis, the other cavernous pulmonary tuberculosis) among 215 contacts in the second ring . Fourteen of the infected persons were centre workers . The bacteria found were not resistant to tuberculostatics . CONCLUSION . Rapid transmission of tuberculosis occurred because of insufficient ventilation and a slow screening procedure . Tuberculosis in refugee centres is a serious problem that needs adequate attention. Proc Natl Acad Sci U S A, 1994 Dec 6, 91(25), 12091 - 5 Detection of conserved segments in proteins: iterative scanning of sequence databases with alignment blocks; Tatusov RL et al.; We describe an approach to analyzing protein sequence databases that, starting from a single uncharacterized sequence or group of related sequences, generates blocks of conserved segments . The procedure involves iterative database scans with an evolving position-dependent weight matrix constructed from a coevolving set of aligned conserved segments . For each iteration, the expected distribution of matrix scores under a random model is used to set a cutoff score for the inclusion of a segment in the next iteration . This cutoff may be calculated to allow the chance inclusion of either a fixed number or a fixed proportion of false positive segments . With sufficiently high cutoff scores, the procedure converged for all alignment blocks studied, with varying numbers of iterations required . Different methods for calculating weight matrices from alignment blocks were compared . The most effective of those tested was a logarithm-of-odds, Bayesian-based approach that used prior residue probabilities calculated from a mixture of Dirichlet distributions . The procedure described was used to detect novel conserved motifs of potential biological importance. Proc Natl Acad Sci U S A, 1994 Dec 6, 91(25), 11816 - 20 Observation of binding and polymerization of Fur repressor onto operator-containing DNA with electron and atomic force microscopes; Le Cam E et al.; The Fur (ferric uptake regulation) protein is a global regulator that, in the presence of Fe2+, represses the expression of a number of iron-acquisition genes and virulence determinants such as toxins . Dark-field electron microscopy of positively stained Fur-DNA complexes in addition to atomic force microscopy allowed direct visualization of Fur interactions with the regulatory regions of aerobactin and hemolysin operons and provided complementary information about the structure of the complexes . According to the DNA used and the protein/DNA ratio, Fur binding to DNA results in partial or total covering of the fragments, indicating that the protein initiates polymerization along the DNA molecules at specific sites . Negative staining of Fur-DNA complexes revealed a well-ordered structure of the polymer suggesting a helical arrangement . Local rigidification of the DNA molecules resulting from Fur binding could be involved in the repression process. Zhonghua Hu Li Za Zhi, 1994 Dec 5, 29(12), 716 - 8 {Air monitoring and method of air sterilization of long-term senile patient wards}; Zhang JH; The air culture was monitored in the wards for aged long-term inpatients and the air sterilizing method was evaluated . The study showed that the natural ventilation was the most practical procedure to reduce the number of bacteria in the air of the wards . The use of Chinese herbal medicine prenaration to eradicate the bacteria in the air by comparison with the ultraviolet light method was proaed to have no sigaificant difference (P > 0.05) . The Chinese herbal medicine method was superior to the ultraviolet light in its safly, reliability, easy operation, moisturizing air, and eradicating the side effect of the ultraviolet light . So it is especially suitable to the wards for aged inpatient. J Biol Chem, 1994 Dec 2, 269(48), 30109 - 12 The 90-kDa heat shock protein is essential for Ah receptor signaling in a yeast expression system; Carver LA et al.; In an effort to provide a more powerful system to study the Ah receptor (AHR) signaling pathway, we expressed the AHR, its dimerization partner ARNT, and a beta-galactosidase (lacZ) reporter gene, driven by two dioxin-responsive enhancers, in the yeast Saccharomyces cerevisiae . In this system, the agonists beta-naphthoflavone and alpha-naphthoflavone induced transcription of the lacZ gene, with EC50 values of 7.9 x 10(-8) and 3.0 x 10(-7) M, respectively, while the nonagonist dexamethasone was without effect . As a first application of this system, we examined the relationship between the 90-kDa heat shock protein (hsp90) and AHR function . To accomplish this in a manner that was independent of the ARNT protein, we constructed a chimeric receptor in which the DNA binding and primary dimerization domains of the AHR were swapped with analogous domains from the LexA protein . Coexpression of this AHR-LexA chimera and a lacZ reporter gene driven by eight LexA operator sites in a yeast strain with regulatable levels of hsp90, yielded pharmacology that closely mirrored that of the AHR/ARNT/dioxin-responsive enhancer system described above, but only when hsp90 levels were held near their wild type levels . When hsp90 levels were reduced to approximately 5% of normal, AHR signaling in response to agonist was completely blocked despite normal cell growth . These results provide the first genetic evidence for the role of hsp90 in AHR signaling and provide the basis for a powerful new system in which to study this pathway. Gene, 1994 Dec 2, 150(1), 123 - 7 Mutations in the linker region of BvgS abolish response to environmental signals for the regulation of the virulence factors in Bordetella pertussis; Manetti R et al.; Expression of virulence factors of Bordetella pertussis is coordinately regulated by the products of the bvg locus, which codes for a sensory protein (BvgS) and a positive regulator of transcription (BvgA), a pair in the family of bacterial 'two-component' regulators . Transcription of the bvg-regulated promoters is repressed by modulating environmental factors such as 50 mM MgSO4, 10 mM nicotinic acid (NA) or low temperature (25 degrees C) . We have isolated a spontaneous mutant (SK170) which expresses virulence genes at either 25 degrees C, or in the presence of 1-5 mM NA, or 10-50 mM MgSO4 . Virulence factors in strain SK170 are still repressed by higher concentrations of NA (10 mM), or by a combination of low temperature (25 degrees C) and one of the other modulating agents . From this strain, we have isolated a second mutant (SK180) that showed constitutive synthesis of the virulence factors under any growth regime . Nucleotide (nt) and deduced amino acid (aa) sequence analysis showed that SK170 contains a substitution at aa570 of BvgS and SK180 contains an additional substitution at aa680 . These substitutions are confined to a 161-aa sequence that links the transmembrane (TM) and kinase domains of BvgS . These mutations also alter the transcriptional autoregulation of the P1 and P2 promoters of the bvg locus . P1, which in the wild-type (wt) strain is repressed by modulating agents, is constitutively active in the mutant strains . On the contrary, P2, which is normally induced by all three modulating agents, is active in strain SK170 only in the presence of MgSO4 or NA, while in strain SK180 this promoter is repressed by modulating agents . The mutants exhibit elevated levels of the BvgA regulatory protein and have a virulent phenotype also in the presence of modulating agents. J Bacteriol, 1994 Dec, 176(24), 7587 - 600 Multiple structural proteins are required for both transcriptional activation and negative autoregulation of Caulobacter crescentus flagellar genes; Ramakrishnan G et al.; The periodic and sequential expression of flagellar (fla) genes in the Caulobacter crescentus cell cycle depends on their organization into levels I to IV of a regulatory hierarchy in which genes at the top of the hierarchy are expressed early in the cell cycle and are required for the later expression of genes below them . In these studies, we have examined the regulatory role of level II fliF operon, which is located near the top of the hierarchy . The last gene in the fliF operon, flbD, encodes a transcriptional factor required for activation of sigma 54-dependent promoters at levels III and IV and negative autoregulation of the level II fliF promoter . We have physically mapped the fliF operon, identified four new genes in the transcription unit, and determined that the organization of these genes is 5'-fliF-fliG-flbE-fliN-flbD-3' . Three of the genes encode homologs of the MS ring protein (FliF) and two switch proteins (FliG and FliN) of enteric bacteria, and the fourth encodes a predicted protein (FlbE) without obvious similarities to known bacterial proteins . We have introduced nonpolar mutations in each of the open reading frames and shown that all of the newly identified genes (fliF, fliG, flbE, and fliN) are required in addition to flbD for activation of the sigma 54-dependent flgK and flbG promoters at level III . In contrast, fliF, fliG, and flbE, but not fliN, are required in addition to flbD for negative autoregulation of the level II fliF promoter . The simplest interpretation of these results is that the requirements of FlbD in transcriptional activation and repression are not identical, and we speculate that FlbD function is subject to dual or overlapping controls . We also discuss the requirement of multiple structural genes for regulation of levels II and III genes and suggest that fla gene expression in C . crescentus may be coupled to two checkpoints in flagellum assembly. Eur J Biochem, 1994 Dec 1, 226(2), 423 - 32 Effects of the Tyr64 substitution on the stability of cytochrome c553, a low oxidoreduction-potential cytochrome from Desulfovibrio vulgaris Hildenborough; Blanchard L et al.; Cytochrome c553 from sulfate-reducing bacteria is a low-oxidoreduction-potential cytochrome . The primary and tertiary structures show notable differences when compared to mitochondrial cytochromes . Tyr64 replacement in cytochrome c553 provides evidence that this residue is not directly involved in the potential modulation but is mostly implicated in the hydrogen-bond network around the heme . While the different variants obtained did not induce drastic structural modifications, they did affect the stability of the protein . This decrease of stability in acidic and alkaline environments was observed by variations in the optical spectra and by mass spectrometry . In addition, the mobility of aromatic side-chain was found to be increased in the mutant proteins as monitored by two-dimensional NMR spectroscopy. Ann Allergy, 1994 Dec, 73(6), 509 - 14 Use of nasal cytology in the diagnosis of occult chronic sinusitis in asthmatic children; Jong CN et al.; BACKGROUND: Chronic sinusitis, in contrast to acute sinusitis, often presents with nonspecific symptoms that may be confused with other disease entities . Due to the cost of computerized tomography and the difficulty in interpreting sinus radiographs in certain children, a search for a simpler screening tool for chronic sinusitis in children was undertaken . OBJECTIVE: This study was undertaken to provide a quantitative comparison between the methods of wax paper blow and Rhinoprobe scraping for nasal cytology in screening for chronic sinusitis while minimizing selection bias . METHODS: Twenty serially selected patients (13 males and seven females) with a mean age of 11 years (range 6-16) were enrolled . Nasal cytology was obtained via two methods: scraping of the turbinate with a Rhinoprobe (Synbiotics Inc.,) and wax paper blow . RESULTS: The results showed that > or = 5 neutrophils per high power field on Rhinoprobe cytology significantly correlated with radiographic sinusitis (P < .05 by Chi-square and P < .056 by Fisher's exact test) . The sensitivity and specificity for > or = 5 neutrophils per high power field were 100% and 53%, respectively . Counts of other nasal cells, such as eosinophils, bacteria, and epithelial cells, did not yield significant correlations with radiographic sinusitis . CONCLUSIONS: We feel that the Rhinoprobe, with criteria of > or = 5 neutrophils per high power field, may be useful as a screen for occult chronic sinusitis in childhood asthma . Confirmation of sinusitis via X-ray is still necessary if neutrophils are present on Rhinoprobe nasal cytology. Dis Colon Rectum, 1994 Dec, 37(12), 1307 - 9 Crohn's disease in the Chinese population . An experience from Hong Kong; Sung JJ et al.; PURPOSE: Crohn's disease was extremely rare among Chinese . We reviewed all cases diagnosed as having Crohn's disease during a five-year period . METHODS: A diagnosis of Crohn's disease was made only if all of the following criteria were fulfilled: 1) clinical symptom(s) and sign(s) compatible with chronic inflammatory bowel disease; 2) exclusion of intestinal infection by repeated stool cultures; 3) macroscopic features of small and/or large intestinal inflammation with skip lesion, stricture, and fistula formation; 4) histologic features of Crohn's disease, i.e., focal lymphoid aggregate, focal cryptitis, and granuloma formation; 5) clinical response to conventional therapy for inflammatory bowel disease . RESULTS: Fifteen ethnic Chinese patients were diagnosed as having Crohn's disease in this period . All patients had colitis, whereas small intestine inflammation was documented in only 47 percent of patients . Extraintestinal manifestations were uncommon except for arthropathy: ankylosing spondylitis (2), sacroiliitis (1), juvenile rheumatoid arthritis (1), and colitic arthritis (1) . The majority of our patients responded to medical therapy . Surgery was undertaken in 33 percent of patients . CONCLUSION: Although there is a general increased incidence of Crohn's disease in the Western world, we too are beginning to see more cases in the Far East . Nevertheless, gastrointestinal infection with bacteria and/or parasites should still be carefully excluded in these countries. Cell Immunol, 1994 Dec, 159(2), 323 - 30 Macrophage response to viruses, protozoa, and fungi: secretory and cellular activities induced in resting unprimed bone marrow-derived mononuclear phagocytes; Keller R et al.; The secretory (tumor necrosis factor, TNF-alpha; nitrite) and cellular response (mitochondrial respiration, TNF-alpha-independent tumoricidal activity) of a pure, lymphocyte-free population of resting, unprimed rat bone marrow-derived mononuclear phagocytes (BMM phi) to direct interaction with viruses, protozoa, and fungi was assessed and compared with that triggered by bacterial agents and interferon-gamma (IFN-gamma) . Viruses (herpes simplex, vaccinia, poliomyelitis, vesicular stomatitis, lymphocytic choriomeningitis, Sendai), protozoa (Trypanosoma brucei, Giardia lamblia), and fungi (Penicillium, Trichosporon, Fusarium, Rhizopus, Aspergillus, Geotrichum species) affected primarily the secretion of TNF-alpha and mitochondrial respiration of BMM phi; their effects on the secretion of nitrite and on tumoricidal activity were at best marginal . Collectively, the macrophage response to viruses, protozoa, and fungi was less varied and less marked than that to bacterial agents (intact organisms, peptidoglycan, lipoteichoic acid, lipopolysaccharide) and IFN-gamma. AJR Am J Roentgenol, 1994 Dec, 163(6), 1333 - 7 Detection of pooled secretions above endotracheal-tube cuffs: value of plain radiographs in sheep cadavers and patients; Greene R et al.; OBJECTIVE . Mechanical ventilation is thought to increase the risk of nosocomial pneumonia by permitting leakage of bacteria-laden gastro-oropharyngeal secretions into the upper airways . The goal of this study was (a) to validate radiographic signs of pooled secretions above endotracheal-tube cuffs (supracuff liquid) in an animal model and (b) to determine whether suctionable pooled supracuff liquid can be identified on bedside radiographs of intubated patients . MATERIALS AND METHODS . Diagnostic criteria for supracuff liquid were initially validated by three radiologists interpreting 162 randomized radiographs made in an intubated sheep cadaver . The primary criteria included (a) replacement of the normal supracuff lucency with liquid opacity and (b) the formation of a sharp interface between the lucency of the upper edge of the cuff below and the liquid above . Graded infusions of 0, 3, 8, 13, and 23 ml of saline were made in triplicate into the space above the cuff, and radiographs were evaluated for the presence or absence of saline . The validated diagnostic criteria were used by two radiologists to estimate the frequency with which pooled liquid was seen on portable chest radiographs of 47 patients undergoing elective short-term postanesthetic mechanical ventilation . RESULTS . In the sheep-cadaver model, the diagnostic criteria for supracuff liquid allowed successful differentiation between no liquid, a small amount of liquid (3-8 ml), and a large amount of liquid (13-23 ml; c2, p < .0001) . In a clinical study, radiographic signs of supracuff liquid were identified in 57% of 47 patients . In a small subset of patients (n = 18), the estimated liquid volume (mean +/- SEM) was calculated to be 7.8 +/- 1.1 ml (range = 2.1-18.4 ml) . CONCLUSIONS . Radiography is a sensitive means of identifying small volumes of supracuff liquid above the inflated cuffs of endotracheal tubes . Potentially contaminating liquid pooled above the cuff of an endotracheal tube can be identified in about half of patients undergoing short-term mechanical ventilation . Our results suggest the suction of the supracuff space may be a reasonable prophylactic maneuver against nosocomial pneumonia . A much larger study is suggested to investigate the actual relation between pooled supracuff liquid and the development of nosocomial pneumonia. J Virol, 1994 Dec, 68(12), 7974 - 85 Targeting of promoters for trans activation by a carboxy-terminal domain of the NS-1 protein of the parvovirus minute virus of mice; Legendre D et al.; The NS-1 gene of the parvovirus minute virus of mice (MVM) (prototype strain, MVMp) was fused in phase with the sequence coding for the DNA-binding domain of the bacterial LexA repressor . The resulting chimeric protein, LexNS-1, was tested for its transcriptional activity by using various target promoters in which multiple LexA operator sequences had been introduced . Under these conditions, NS-1 was shown to stimulate gene expression driven by the modified long terminal repeat promoters (from the retroviruses mouse mammary tumor virus and Rous sarcoma virus) and P38 promoter (from MVMp), indicating that the NS-1 protein is a potent transcriptional activator . It is noteworthy that in the absence of LexA operator-mediated targeting, the genuine mouse mammary tumor virus and Rous sarcoma virus promoters were inhibited by NS-1 . Together these data strongly suggest that NS-1 contains an activating region able to induce promoters with which this protein interacts but also to repress transcription from nonrecognized promoters by a squelching mechanism similar to that described for other activators . Deletion mutant analysis led to the identification of an NS-1 domain that exhibited an activating potential comparable to that of the whole polypeptide when fused to the DNA-binding region of LexA . This domain is localized in the carboxy-terminal part of NS-1 and corresponds to one of the two regions previously found to be responsible for toxicity . These results argue for the involvement of the regulatory functions of NS-1 in the cytopathic effect of this parvovirus product. J Exp Med, 1994 Dec 1, 180(6), 2359 - 64 The germfree state prevents development of gut and joint inflammatory disease in HLA-B27 transgenic rats; Taurog JD et al.; A number of inflammatory disease states occur with greatly increased frequency in individuals inheriting the human major histocompatibility complex class I allele HLA-B27 . In a minority of cases, namely those with B27-associated reactive arthritis, there is good evidence that the disease state is triggered by infection with an enteric or genitourinary bacterial pathogen . For the majority of B27-associated disease, no definite pathogenetic role for bacteria has been established . However, in these latter cases intestinal inflammation can often be demonstrated, and it sometimes occupies a major part of the clinical picture . Rats transgenic for B27 are known to develop a disorder resembling B27-associated human disease, with prominent intestinal, joint, skin, and male genital inflammatory lesions . We report here that B27 transgenic rats raised in a germfree environment do not develop inflammatory intestinal or peripheral joint disease, whereas the skin and genital inflammatory lesions are unaffected by the germfree state . These findings support the concept that gut and joint inflammation are pathogenetically closely related, and they provide direct evidence that the commensal gut flora play an important role in the pathogenesis of B27-associated gut and joint inflammation. J Bacteriol, 1994 Dec, 176(23), 7409 - 11 Transcriptional and translational analysis of ferredoxin and flavodoxin under iron and nitrogen stress in Anabaena sp . strain PCC 7120; Razquin P et al.; In Anabaena sp . strain PCC 7120, vegetative cell ferredoxin synthesis under iron starvation was repressed 25-fold, whereas heterocyst ferredoxin synthesis decreased only 2.8-fold . Induction of flavodoxin under iron depletion was independent of the availability of combined nitrogen . Under iron stress but in the presence of combined nitrogen, fdxH and nifH genes were transcriptionally active; although excision of the 11-kb element seemed to be completed, nitrogenase activity and the fdxH gene product were not detectable. Infect Immun, 1994 Dec, 62(12), 5528 - 37 Invasion and intracellular survival of Bordetella bronchiseptica in mouse dendritic cells; Guzman CA et al.; We have studied the interaction between the respiratory pathogen Bordetella bronchiseptica and murine spleen dendritic cells, important antigen-presenting cells that are found in the airway epithelium . Wild-type B . bronchiseptica 5376 attached very efficiently to dendritic cells, whereas the bvg mutant ATCC 10580, wild-type strain BB7865, and its spontaneous delta bvgS mutant BB7866 bound less efficiently . However, all tested B . bronchiseptica strains were able to invade dendritic cells and survive intracellularly for at least 72 h . These results suggest that bvg-independent or bvg-downregulated products are involved in the uptake and intracellular survival . Transmission electron microscopic analysis revealed that bacteria grew and replicated intracellularly and were present in typical phagosomes, which fused with lysosomes during the initial infection period . However, in later infection stages some bacteria seemed to escape into an unfused endocytic compartment, where individual bacteria were tightly surrounded by a membrane . The in vitro interaction of B . bronchiseptica with dendritic cells reported here may be relevant to natural infections caused by this organism that lead to chronicity or an altered immune response. J Hosp Infect, 1994 Dec, 28(4), 245 - 8 Ultraviolet radiation and the control of airborne contamination in the operating room; Lidwell OM; Ultraviolet irradiation has been employed in operating rooms for more than half a century in attempts to reduce airborne bacterial contamination . Safety considerations have limited its intensity to 25-30 mu w cm-2 and at this level no more than a fourfold reduction has resulted . In recent studies intensities up to 300 mu w cm-2 have been used without untoward effects, and, at the highest intensity, contamination as low as that obtained with ultraclean air ventilation systems was obtained . However this was only achieved in an operating room where the level of airborne contamination before the introduction of the radiation was already much lower (around one-fifth) than that usually observed, and the reduction attributable to the radiation was still only about 12-fold. J Bacteriol, 1994 Dec, 176(23), 7395 - 7 Transformation of Chlorobium limicola by a plasmid that confers the ability to utilize thiosulfate; Mendez-Alvarez S et al.; A modified transformation method for Chlorobium limicola confirms the role of an endogenous plasmid in thiosulfate metabolism . The plasmid was present in two forma specialis thiosulfatophilum (Tio+) strains and absent in one non-thiosulfate-utilizing (Tio-) strain . The plasmid (size, 14 kb) was transferred by transformation from Tio- to Tio+ . The chromosomal restriction patterns, analyzed by pulsed-field gel electrophoresis, were used to distinguish between the different strains. Trends Microbiol, 1994 Dec, 2(12), 465 - 9 Restriction enzymes in cells, not eppendorfs; King G et al.; Restriction enzymes are essential reagents to molecular biologists, but their relevance to bacterial populations is less obvious . Most bacteria encode restriction and modification systems and these are commonly considered to be a barrier to phage infection . Current evidence also supports a more general role for them in genetic recombination. Semin Respir Infect, 1994 Dec, 9(4), 228 - 31 Effect of route of feeding on the incidence of septic complications in critically ill patients; Minard G et al.; The increased risk of septic complications accompanying severe illness and injury is compounded by the presence of malnutrition . Total parenteral nutrition (TPN) has been used extensively to prevent or rectify this problem . Although enteral nutrition is frequently more difficult to administer, a growing body of laboratory and clinical research shows a significant reduction in the incidence of secondary infection with its use . The mechanism proposed is that the enteral route helps maintain the gut barrier, decreasing passage of bacteria and other toxins . Translocation of these products has been implicated as a cause of nosocomial infection and organ failure . Therefore, when possible, the use of the enteral route of nutrition should be part of the overall approach to the care of the critically ill or injured patient. Kansenshogaku Zasshi, 1994 Dec, 68(12), 1479 - 83 {The role of viral infection in adults with bronchopulmonary infections}; Konishi M et al.; We isolated the virus from the respiratory specimens of 16 patients (7.9%) when we investigated the bronchopulmonary infections of 203 adult patients by the viral isolation method for the diagnosis of the viral infection and transtracheal aspiration (TTA) for taking directly the specimen from the lower respiratory tract . The bronchopulmonary infections, isolated the virus, consisted of acute bronchitis (3 cases), pneumonia (8 cases) and the acute exacerbation of chronic lower respiratory infection (5 cases) . The positive viral isolation consisted of Rhinovirus (11 strains), Herpes simplex virus I (3 strains), Parainfluenza virus III (1 strain) and Respiratory syncytial virus (1 stain) . There were 9 transtracheal aspirates, 8 pharyngeal swabs and 5 nasal washings in the positive viral specimens . We experienced 5 cases who had the same virus isolated from both the upper respiratory tract and the lower respiratory tract . H . influenzae, P . aeruginosa and other bacteria were found in 14 of the 16 TTA cultures . These results suggest that the role of the viral infection is important either independently or relating the bacterial infection in the adult bronchopulmonary infections. J Clin Gastroenterol, 1994 Dec, 19(4), 288 - 91 Cryptosporidium . A cause of gastroenteritis in preschool children in Jordan; Nimri LF et al.; In this case-control study, we investigated the role of Cryptosporidium in gastroenteritis in children < 6 years old . Six hundred fresh stool specimens were examined for various pathogenic parasites, bacteria, and rotaviruses . Wet-mount preparations, formaline-ether concentrations, and Sheather's floatation techniques were used to recover the parasite oocysts . Permanent stained slides using acid-fast stain and trichrome stains were prepared . Of 300 children with gastroenteritis symptoms, 20 (6.7%) had Cryptosporidium oocysts; seven of the 20 had concomitant infections so they were excluded from the counts . This infection rate is significantly different (Z = 2; p < 0.05) from that found in the control group (1.7%) of children who reported no symptoms . The most frequent symptoms reported beside diarrhea were abdominal pain, cramps, anorexia, nausea, vomiting, and fatigue . Contaminated drinking water is suspected to be the source of infection; other possible factors are discussed. Immunology, 1994 Dec, 83(4), 659 - 64 Recombinant L7/L12 ribosomal protein and gamma-irradiated Brucella abortus induce a T-helper 1 subset response from murine CD4+ T cells; Oliveira SC et al.; Immunity to Brucella abortus crucially depends on antigen (Ag)-specific T-cell mediated activation of macrophages, which are the major effectors of cell-mediated killing of this organism . Ribosomal preparations have been used as vaccines against several pathogens, including B . abortus, conferring a high degree of protection . In the present study, we have examined the pattern of T-helper (Th) cell response from infected BALB/c mice after in vitro stimulation with recombinant (r) L7/L12 ribosomal protein or gamma-irradiated B . abortus . In addition to Ag-specific proliferation, CD4+ T cells were tested for interleukin-2 (IL-2), IL-4 and interferon-gamma (IFN-gamma) mRNA expression and secretion . Detection of cytokine transcripts and secreted cytokines was performed using reverse transcriptase (RT)-polymerase chain reaction (PCR) and specific ELISA assays . Primed CD4+ T cells proliferated to the recombinant protein or whole B . abortus . The functional cytokine profile of the proliferating cells was typical of a Th1 cell phenotype, as we detected transcripts for IL-2 and IFN-gamma but not IL-4 . Among the cytokines analysed, only IFN-gamma produced in the Th cell culture supernatants was detected by ELISA when bacteria or recombinant protein were used . Thus, rL7/L12 ribosomal protein and gamma-irradiated B . abortus preferentially stimulated IFN-gamma-producing Th1 cells after in vitro stimulation . The results of this study provide for the first time an explanation of why ribosomal vaccines may protect against intracellular infections, and an experimental basis for identifying polypeptides from a pathogen which stimulates the desired cytokine profile and Th cell response crucial for the design of genetically engineered candidate vaccines. Oral Microbiol Immunol, 1994 Dec, 9(6), 372 - 5 Initial catabolism of sorbitol in Actinomyces naeslundii and Actinomyces viscosus; Kalfas S et al.; The initial steps of sorbitol catabolism were studied in 4 strains of Actinomyces naeslundii and Actinomyces viscosus that had been isolated from human dental plaque . Cell-free extracts were prepared from cells grown in the presence of either sorbitol, xylitol or glucose . The extracts from all strains grown on sorbitol had nicotinamide adenine dinucleotide-linked dehydrogenase activities for sorbitol and xylitol and reduced nicotinamide adenine dinucleotide-linked reductase activities for fructose and xylulose . Two of the strains also exhibited these activities when grown in the presence of xylitol, and all glucose-grown cells lacked them . The results indicate that sorbitol metabolism in oral actinomyces involve oxidation of sorbitol to fructose by an inducible enzyme, nicotinamide adenine dinucleotide-linked sorbitol dehydrogenase . This step is followed by the phosphorylation of fructose with guanosine triphosphate as a main phosphoryl donor . Thus, the initial catabolic pathway of sorbitol in A . naeslundii and A . viscosus is different from those described for other oral bacteria. Nippon Hinyokika Gakkai Zasshi, 1994 Dec, 85(12), 1709 - 16 {Clinical study on female patients with bladder irritability--comparison between cystitis and urethral syndrome}; Koroku M et al.; We analysed the clinical background of female who had no underlying disease but consulted out urology outpatient clinic with a chief complaint of symptoms of bladder irritation . Overally, 50.1% (102/200 cases) of the cas