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Sample records for chlamydophila pneumoniae protein

  1. Infections caused by Chlamydophila pneumoniae.

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    Choroszy-Król, Irena; Frej-Mądrzak, Magdalena; Hober, Martyna; Sarowska, Jolanta; Jama-Kmiecik, Agnieszka

    2014-01-01

    High affinity to the epithelial lining of the airways makes Chlamydophila pneumoniae a common etiological agent of respiratory tract infections (RTI). It causes among others: pharyngitis, tracheitis, sinusitis, otitis media, bronchitis and bronchiolitis, and pneumonia. It is estimated that Chlamydophila pneumoniae infection is responsible for about 20% of lower respiratory tract infections. Chlamydophila pneumoniae infection may play an important role in the pathogenesis and course of bronchial asthma. The recent results indicate that Chlamydophila pneumoniae infection may be a factor responsible for 4-16% of COPD (Chronic obstructive pulmonary disease) exacerbations. A relationship of chlamydial infection with atherosclerosis raises huge interest. A connection of Chlamydophila pneumoniae infection with other non-communicable diseases such as lung cancer, arthritis, Alzheimer's disease, multiple sclerosis, sarcoidosis and erythema nodosum is also recognized, although the role of these bacteria has not been fully understood in any of the listed diseases.

  2. Chlamydophila pneumoniae Infection Induced Nodular Vasculitis

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    H. Sakuma

    2011-12-01

    Full Text Available We present the case history of a 48-year-old male patient with Chlamydophila (Chlamydia pneumoniae who developed a nodular vasculitis. He developed a cutaneous vasculitis with the onset of respiratory symptoms. The diagnosis of Chlamydophila pneumoniae infection was based on serology. Since this infection is very common in our population, although often asymptomatic, it should be systematically considered as a causative agent of nodular vasculitis.

  3. Enzymatic characterization of Chlamydophila pneumoniae phospholipase D.

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    Mancini, Fabiola; Ciervo, Alessandra

    2015-01-01

    Chlamydophila pneumoniae, an aetiological agent of respiratory infection, is also thought to play an immuno-pathogenetic role in atherosclerosis by contributing to inflammation and plaque instability. Phospholipase D (PLD) is an enzyme involved in lipid metabolism and may have a direct or indirect impact on virulence and the inflammatory response. Some aspects of the developmental cycle of C. pneumoniae suggest a direct implication of its PLD (CpPLD) in the pathogenesis, specifically by affecting the regulation of lipid metabolism and lipid exchange between C. pneumoniae and host cells. Our previous studies disclosed a specific anti-CpPLD antibody response in patients with acute coronary syndromes chronically infected with C. pneumoniae, and demonstrated that this antigen is a factor able to drive the inflammatory process in atherosclerosis. Due to the intriguing aspects of the CpPLD, the present study investigated CpPLD enzymatic activity of the protein and the two domains that include one HKD motif each polypeptide. Our results showed that CpPLD was able to synthesize the cardiolipin (CL) but unable to hydrolyze phospholipids. It was also observed that each single HKD motif has an independent CL synthetase activity. This enzymatic activity of CpPLD could be important in the inflammatory process within the atherothrombotic events.

  4. Biomarker Candidates of Chlamydophila pneumoniae Proteins and Protein Fragments Identified by Affinity-Proteomics Using FTICR-MS and LC-MS/MS

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    Susnea, Iuliana; Bunk, Sebastian; Wendel, Albrecht; Hermann, Corinna; Przybylski, Michael

    2011-04-01

    We report here an affinity-proteomics approach that combines 2D-gel electrophoresis and immunoblotting with high performance mass spectrometry to the identification of both full length protein antigens and antigenic fragments of Chlamydophila pneumoniae (C. pneumoniae). The present affinity-mass spectrometry approach effectively utilized high resolution FTICR mass spectrometry and LC-tandem-MS for protein identification, and enabled the identification of several new highly antigenic C. pneumoniae proteins that were not hitherto reported or previously detected only in other Chlamydia species, such as Chlamydia trachomatis. Moreover, high resolution affinity-MS provided the identification of several neo-antigenic protein fragments containing N- and C-terminal, and central domains such as fragments of the membrane protein Pmp21 and the secreted chlamydial proteasome-like factor (Cpaf), representing specific biomarker candidates.

  5. Chlamydophila pneumoniae infection and cardiovascular disease

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    Rajnish Joshi

    2013-01-01

    Full Text Available Atherosclerosis is a multifactorial vascular inflammatory process; however, the inciting cause for inflammation remains unclear. Two decades ago, Chlamydophila pneumoniae (formerly Chlamydia pneumoniae infection was proposed as a putative etiologic agent. We performed a PubMed search using the keywords Chlamydia and atherosclerosis in a Boolean query to identify published studies on C. pneumoniae and its role in atherogenesis, and to understand research interest in this topic. We found 1,652 published articles on this topic between 1991 and 2011. We analyzed relevant published studies and found various serological, molecular, and animal modeling studies in the early period. Encouraged by positive results from these studies, more than a dozen antibiotic clinical-trials were subsequently conducted, which did not find clinical benefits of anti-Chlamydophila drug therapy. While many researchers believe that the organism is still important, negative clinical trials had a similar impact on overall research interest. With many novel mechanisms identified for atherogenesis, there is a need for newer paradigms in Chlamydophila-atherosclerosis research.

  6. Detection of Chlamydophila pneumoniae antigens in patients with chronic cough.

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    Choroszy-Krol, Irena; Frej-Madrzak, Magdalena; Jama-Kmiecik, Agnieszka; Sarowska, Jolanta; Serek, Pawel; Pirogowicz, Iwona; Nittner-Marszalska, Marita

    2013-01-01

    The aim of this study was to analyze the rate of Chlamydophila pneumoniae infection in adults with symptoms of chronic cough. The study was conducted in 83 hospitalized patients aged 18-67 suffering of chronic cough. The control group consisted of 20 healthy age-matched subjects without any respiratory symptoms. Bacteriological tests on the presence of Chlamydophila pneumoniae antigen were performed in throat swabs by indirect immunofluorescence technique using monoclonal antibodies labeled with fluorescein isothiocyanate. The rate of Chlamydophila infected patients was examined in relation to age and gender. The Chlamydophila pneumoniae antigen was detected in 15 (18 %) out of the 83 patients; about equally in both genders. Furthermore, we found that the patients aged 28-37 constituted the age group that most frequently tested positive for Chlamydophila pneumoniae. Unraveling the presence of Chlamydia infection in chronic cough patients enables to introduce a timely implementation of effective therapy and thus can prevent distant complications.

  7. A Multiplex PCR for Detection of Mycoplasma pneumoniae, Chlamydophila pneumoniae, Legionella pneumophila, and Bordetella pertussis in Clinical Specimens

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    2007-11-02

    NAVAL HEALTH RESEARCH CENTER A MULTIPLEX PCR FOR DETECTION OF Mycoplasma pneumoniae, Chlamydophila pneumoniae, Legionella pneumophila, AND Bordetella...5300 2 A Multiplex PCR for Detection of Mycoplasma pneumoniae, Chlamydophila pneumoniae, Legionella pneumophila, and Bordetella pertussis in Clinical... Chlamydophila pneumoniae, Legionella pneumophila, and Bordetella pertussis in uncultured patient specimens. These organisms cause similar symptomologies

  8. Association of Alzheimer's disease and Chlamydophila pneumoniae.

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    Stallings, Tiffany L

    2008-06-01

    This paper critically reviews the association of infection by Chlamydophila pneumoniae (C. pneumoniae) and Alzheimer's disease (AD). The aging population has increased interest in finding the cause of AD, but studies have yielded contradictory results that are likely due to varying diagnostic tools and different uses of diagnostic tests. Knowledge of AD's characteristics, risk factors, and hypothesized etiologies has expanded since Alois Alzheimer's initial description of AD. Epidemiologic and projection studies provide incidence estimates of AD through a two-stage method: (1) primary diagnosis of dementia by cognitive testing such as Mini-Mental State Examination (MMSE), and (2) clinical diagnosis of AD through criteria such as National Institute of Neurological and Communicative Diseases and Stroke/Alzheimer's Disease and Related Disorders Association (NINCDS-ADRDA). Cross-sectional studies yield prevalence estimates of infection by C. pneumoniae by detecting immunoglobulins through laboratory tests such as microimmunofluorescence (MIF). Studies examining the association of C. pneumoniae and AD are limited, but brain autopsy provides information about presence, proximity to areas associated with AD, and bacterial load. Standardization of diagnostic techniques would allow for better comparability of studies, but uncertainty about the best method of diagnosis of infection by C. pneumoniae and AD may call for revised or novel diagnostic tools.

  9. Three cases of atypical pneumonia caused by Chlamydophila psittaci.

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    Chau, S; Tso, Eugene Y K; Leung, W S; Fung, Kitty S C

    2015-06-01

    Psittacosis is a zoonotic disease caused by Chlamydophila psittaci. The most common presentation is atypical pneumonia. Three cases of pneumonia of varying severity due to psittacosis are described. All patients had a history of avian contact. The diagnosis was confirmed by molecular detection of Chlamydophila psittaci in respiratory specimens. The cases showed good recovery with doxycycline treatment. Increased awareness of psittacosis can shorten diagnostic delay and improve patient outcomes.

  10. Liver granulomatosis: a case of Chlamydophila pneumoniae infection.

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    Yildiz, H; Wieërs, G; Yombi, J C; Marot, J C

    2015-02-01

    An 18-year-old man was referred to the Internal Medicine ward because of a 2-week history of intermittent high fever, weight loss and cough. Clinical examination revealed hepato-splenomegaly and multiple lymph nodes swelling while laboratory tests showed elevated C-reactive protein, gamma glutamyl transferase and lactate dehydrogenase. All serologic testes for auto-immune antibodies, viruses and bacteria were negative except for Chlamydophila pneumoniae. An 18-FDG PET computed tomography scanner showed hypermetabolism in the liver, spleen and lymph nodes. We therefore conducted a liver biopsy that demonstrated non-necrotizing granulomas. We conclude to a C. pneumoniae infection associated with a granulomatous hepatitis. After treatment with Doxyciclin the patient had no more fever, hepatosplenomegaly resolved and blood testes normalized. This case report is to our knowledge the first report of a granulomatous hepatitis associated with C. pneumoniae respiratory infection.

  11. A novel inhibitor of Chlamydophila pneumoniae protein kinase D (PknD inhibits phosphorylation of CdsD and suppresses bacterial replication

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    Bulir David C

    2009-10-01

    Full Text Available Abstract Background We have shown previously that Chlamydophila pneumoniae contains a dual-specific Ser/Thr protein kinase that phosphorylates CdsD, a structural component of the type III secretion apparatus. To further study the role of PknD in growth and development we sought to identify a PknD inhibitor to determine whether PknD activity is required for replication. Results Using an in vitro kinase assay we screened 80 known eukaryotic protein kinase inhibitors for activity against PknD and identified a 3'-pyridyl oxindole compound that inhibited PknD autophosphorylation and phosphorylation of CdsD. The PknD inhibitor significantly retarded the growth rate of C. pneumoniae as evidenced by the presence of very small inclusions with a reduced number of bacteria as seen by electron microscopy. These inclusions contained the normal replicative forms including elementary bodies (EB, intermediate bodies (IB and reticulate bodies (RB, but lacked persistent bodies (PB, indicating that induction of persistence was not the cause of reduced chlamydial growth. Blind passage of C. pneumoniae grown in the presence of this PknD inhibitor for 72 or 84 hr failed to produce inclusions, suggesting this compound blocks an essential step in the production of infectious chlamydial EB. The compound was not toxic to HeLa cells, did not block activation of the MEK/ERK pathway required for chlamydial invasion and did not block intracellular replication of either Chlamydia trachomatis serovar D or Salmonella enterica sv. Typhimurium suggesting that the inhibitory effect of the compound is specific for C. pneumoniae. Conclusion We have identified a 3'-pyridyl oxindole compound that inhibits the in vitro kinase activity of C. pneumoniae PknD and inhibits the growth and production of infectious C. pneumoniae progeny in HeLa cells. Together, these results suggest that PknD may play a key role in the developmental cycle of C. pneumoniae.

  12. Pneumonia Outbreak Caused by Chlamydophila pneumoniae Among US Air Force Academy Cadets, Colorado, USA

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    2015-06-01

    Article 3. DATES COVERED (From – To) Sep 2013 – May 2014 4. TITLE AND SUBTITLE Pneumonia Outbreak Caused by Chlamydophila pneumoniae among US Air Force...October 2013–May 2014, there were 102 cases of pneumonia diagnosed in US Air Force Academy cadets. A total of 73% of tested nasal washes contained...Chlamydophila pneumoniae . This agent can be considered to be present on campus settings during outbreaks with numerous, seemingly disconnected cases of

  13. Preparation and analysis of immunocompetence of recombinant fusion protein of the immunodominant region in chlamydial protease-like activity factor from Chlamydophila pneumoniae and its application in serodiagnosis

    Institute of Scientific and Technical Information of China (English)

    JIANG HUA ZHENG; YI MOU WU; TAO DING; LI LI CHEN; JIA QIANG LIU; SHUANG QUAN LIU

    2007-01-01

    To clone the gene coding the immunodominant region in the chlamydial protease-like activity factor (CPAF) from Chlamydophila pneumoniae, to analyze immunocompetence of the expressed protein,and to evaluate its value in serodiagnosis, the CPAF immunodominant region gene was amplified, ligated into a pGEX6p-2 vector, and then the expressed recombinant protein was purified with glutathione Stransferase (GST) agarose gel FF after renaturation, then identified by SDS-PAGE and Western blot. A new indirect ELISA was developed with the purified protein as coating antigen. The immunogenicity of the recombinant protein was evaluated by immunization to New Zealand rabbits, and its immunoreactivity was analyzed by reacting with anti-C, pneumoniae antibody. 300 clinical sera samples were respectively detected by microimmunofluorescence (MIF) as reference method and the indirect ELISA, and the difference between the two methods was analyzed. Cross-reactivity against Chlamydia trachomatis was investigated with the indirect ELISA to detect anti-C, trachomatis positive antisera. The results indicated that a 51.3 kDa recombinant protein was obtained. Western blot assay proved that the recombinant protein could merely specifically react with human anti- C. pneumoniae antisera. The titers of the specific IgG antibodies in the immunized New Zealand rabbits were above 1 : 16 000. Anti- C. pneumoniae IgG positive and negative reference sera were detected with the indirect ELISA, and the concordance rate of negative and positive results were both 100% (40/40). The sensitivity and specificity of the indirect ELISA in comparison with MIF were 93.8% (45/48) and 100% (252/252) separately by detecting 300 clinical sera samples, and the concordance rate between the two methods was 99.0%. No cross reaction against C. trachomatis was found with the indirect ELISA to detect anti-C, trachomatis positive antisera. In conclusion, the prepared recombinant protein of the CPAF immunodominant region

  14. Immunogenicity of Cpn0425 and its localization in cells infected with Chlamydophila pneumoniae.

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    Liu, Liangzhuan; You, Xiaoxing; Chen, Liesong; Zeng, Yanhua; Tang, Shuangyang; Yu, Minjun; Wu, Yimou; Xhen, Xi

    2012-12-01

    The present study aimed to determine the intracellular localization of Cpn0425 in Chlamydophila pneumoniae-infected cells. The recombinant plasmid pGEX-6p/Cpn0425 was transformed into E.coli Bl21 cells to express the fusion protein. Following purification with glutathione S-transferase (GST) resin chromatography, the Cpn0425 fusion protein was used to induce immunity in mice to develop monoclonal and polyclonal antibodies, which were subsequently used to localize the endogenous Cpn0425 protein by indirect immunofluorescence assay (IFA). ELISA was used to determine the immunogenicity of the Cpn0425 plasmid protein by recognizing the pool sera of patients infected with Chlamydia trachomatis and the pool sera of mice immunized with the Cpn0425 fusion protein. The Cpn0425 gene was expressed as the GST-Cpn0425 fusion protein in E. coli and its antibody was prepared by immunizing mice with the fusion protein. An anti-GST-Cpn0425 antibody was used to localize the protein in cells infected with Chlamydophila pneumoniae AR-39 using an IFA. The anti-GST-CT058 antibody detected an inclusion signal in the IFA. Cpn0425 protein strongly reacted with antiserum. Although Cpn0425 protein is not a secreted protein, it has good immunogenicity. Therefore, this protein may be useful for developing vaccines against Chlamydophila pneumoniae infection.

  15. Molecular evidence of Chlamydophila pneumoniae infection in reptiles in Argentina.

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    Frutos, María C; Monetti, Marina S; Ré, Viviana E; Cuffini, Cecilia G

    2014-01-01

    In the central area of Argentina, the epidemiological and molecular characteristics of Chlamydophila pneumoniae infections in reptiles are still unknown. A nested polymerase chain reaction of the rpoB gene was used to detect C. pneumoniae in cloacal swab samples from 19 reptiles at a recreational area. Eleven (57.89%) reptiles were positive; the sequencing and phylogenetic analysis confirmed the presence of this bacterium. Neither C. pneumoniae DNA in the caregivers pharynges nor IgM antibodies anti-C. pneumoniae in their serum samples were detected; however, caregivers presented very high titers of IgG anti-C. pneumoniae. The detection of C. pneumoniae DNA in reptiles demonstrated the circulation of this agent in the recreational area and could be responsible for the exacerbated immune response of the personnel handling the reptiles, which suggests a potential zoonotic cycle. This is the first report of the detection of C. pneumoniae in reptiles in Argentina.

  16. Mycoplasma Pneumoniae and Chlamydophila pneumoniae in hospitalized children with bronchiolitis.

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    Zirakishvili, D; Chkhaidze, I; Barnabishvili, N

    2015-03-01

    Bronchiolitis is an acute lower respiratory tract infection in early childhood caused mainly by different viruses. Etiology of bronchiolitis have been studied in different environments and populations. Respiratory syncytial virus (RSV), human Metapneumovirus (hMPV), human Bocavirus (hBoV), human Rhinoviruses (hRV) have consistently been shown to predominate. Few studies however have attempted to determine whether other pathogens, particularly Mycoplasma Pneumoniae (MP) and Chlamydophila pneumoniae (CP), are associated with bronchiolitis in children under 2 years of age. The aim of this study was to determine the prevalence and clinical features of MP and CP in children under the age of 2 years presenting to the Iashvili Central Children Hospital in Tbilisi with various severities and clinical manifestations of bronchiolitis. Acute and convalescent serum samples were tested by ELISA for IgM and IgG antibodies to RSV, CP and MP.37 children under two years of age were studied. In 19 patients out of 37 (51.35%) etiological diagnosis were established and in 18 patients (48.65%) no pathogens were found. 11 patients (29.72%) had either CP or MP and 8 patients (21.62%) had RSV. Children infected with CP and MP had less severe bronchiolitis than those infected with RSV. Co-infection was not associated with disease severity. There were no statistically significant differences between groups with respect to length of hospital stay. Our study underlines the importance of atypical bacterial pathogens in acute bronchiolitis in children under 2 years and highlights the complex epidemiology and clinical features of these pathogens in this age group.

  17. Antibody responses of Chlamydophila pneumoniae pneumonia: Why is the diagnosis of C. pneumoniae pneumonia difficult?

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    Miyashita, Naoyuki; Kawai, Yasuhiro; Tanaka, Takaaki; Akaike, Hiroto; Teranishi, Hideto; Wakabayashi, Tokio; Nakano, Takashi; Ouchi, Kazunobu; Okimoto, Niro

    2015-07-01

    The ELNAS Plate Chlamydophila pneumoniae commercial test kit for the detection of anti-C. pneumoniae-specific immunoglobulin M (IgM), IgA and IgG antibodies has become available in Japan recently. To determine the optimum serum collection point for the ELNAS plate in the diagnosis of C. pneumoniae pneumonia, we analyzed the kinetics of the antibody response in patients with laboratory-confirmed C. pneumoniae pneumonia. We enrolled five C. pneumoniae pneumonia cases and collected sera from patients for several months. The kinetics of the IgM and IgG antibody responses were similar among the five patients. Significant increases in IgM and IgG antibody titer between paired sera were observed in all patients. IgM antibodies appeared approximately 2-3 weeks after the onset of illness, reached a peak after 4-5 weeks, and were generally undetectable after 3-5 months. IgG antibodies developed slowly for the first 30 days and reached a plateau approximately 3-4 months after the onset of illness. The kinetics of IgA antibody responses were different among the five patients, and significant increases in IgA antibody titer between paired sera were observed in only two patients. Although the sample size was small, the best serum collection time seemed to be approximately 3-6 weeks after onset of illness when using a single serum sample for the detection of IgM antibodies. Paired sera samples should be obtained at least 4 weeks apart. IgA antibody analysis using ELNAS may not be a useful marker for acute C. pneumoniae pneumonia.

  18. Atherosclerosis Induced by Chlamydophila pneumoniae: A Controversial Theory

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    Hamidreza Honarmand

    2013-01-01

    Full Text Available More than a century ago, inflammation and infection were considered to have atherogenic effects. The old idea that coronary heart disease (CHD possibly has an infectious etiology has only reemerged in recent years. Atherosclerosis is the main pathological process involved in CHD and is, logically, the first place to look for infectious etiology. The process of atherosclerosis itself provides the first hints of potential infectious cause. Smooth muscle proliferation, with subsequent intimal thickening, luminal narrowing, and endothelial degeneration, constitutes the natural history of atherosclerosis, being with the severity and speed of these changes. Both viral and bacterial pathogens have been proposed to be associated with the inflammatory changes found in atherosclerosis. Recently, Chlamydophila pneumoniae (C. pneumoniae has been implicated as a possible etiologic agent of coronary artery disease and atherosclerosis. New evidence which supports a role for C. pneumoniae in the pathogenesis of atherosclerosis has emerged. C. pneumoniae has been detected in atherosclerotic arteries by several techniques, and the organism has been isolated from both coronary and carotid atheromas. Recent animal models have suggested that C. pneumoniae is capable of inducing atherosclerosis in both rabbit and mouse models of atherosclerosis. Furthermore, human clinical treatment studies which examined the use of antichlamydial macrolide antibiotics in patients with coronary atherosclerosis have been carried out. The causal relationship has not yet been proven, but ongoing large intervention trials and research on pathogenetic mechanisms may lead to the use of antimicrobial agents in the treatment of CHD in the future.

  19. Genomic screening for Chlamydophila pneumoniae-specific antigens using serum samples from patients with primary infection.

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    Yasui, Yumiko; Yanatori, Izumi; Kawai, Yasuhiro; Miura, Koshiro; Suminami, Yoshinori; Hirota, Tomomitsu; Tamari, Mayumi; Ouchi, Kazunobu; Kishi, Fumio

    2012-04-01

    Chlamydophila pneumoniae, an obligate intracellular human pathogen, causes respiratory tract infections. The most common techniques used for the serological diagnosis of C. pneumoniae infections are microimmunofluorescence tests and commercial serological ELISA tests; these are based on the detection of antibodies against whole chlamydial elementary bodies and lipopolysaccharide/outer membrane protein, respectively. Identification of more specific and highly immunodominant antigens is essential for the development of new serodiagnostic assays. To identify novel specific antigens from C. pneumoniae, we screened 455 genes with unknown function in the genome of C. pneumoniae J138. Extracts of Saccharomyces cerevisiae cells expressing GFP-tagged C. pneumoniae proteins were subjected to Western blot analysis using serum samples from C. pneumoniae-infected patients as the primary antibodies. From this comprehensive analysis, 58 clones expressing C. pneumoniae open reading frames, including hypothetical proteins, were identified as antigens. These results have provided useful information for the development of new serological tools for the diagnosis for C. pneumoniae infections and for the development of vaccines in future.

  20. Chlamydophila pneumoniae induces a sustained airway hyperresponsiveness and inflammation in mice

    NARCIS (Netherlands)

    F. Blasi (Francesco); S. Aliberti (Stefano); L. Allegra (Luigi); G. Piatti (Gioia); P. Tarsia (Paolo); J.M. Ossewaarde (Jacobus); V. Verweij (Vivienne); F.P. Nijkamp (Frans); G. Folkerts (Gert)

    2007-01-01

    textabstractBackground: It has been reported that Chlamydophila (C.) pneumoniae is involved in the initiation and promotion of asthma and chronic obstructive pulmonary diseases (COPD). Surprisingly, the effect of C. pneumoniae on airway function has never been investigated.Methods: In this study, mi

  1. Ubiquinone (Coenzyme Q) Biosynthesis in Chlamydophila pneumoniae AR39: Identification of the ubiD Gene

    Institute of Scientific and Technical Information of China (English)

    Jun LIU; Jian-Hua LIU

    2006-01-01

    Ubiquinone is an essential electron carrier in prokaryotes. Ubiquinone biosynthesis involves at least nine reactions in Escherichia coli. 3-octaprenyl-4-hydroxybenzoate decarboxylase (UbiD) is an important enzyme on the pathway and deletion of the ubiD gene in E. coli gives rise to ubiquinone deficiency in vivo.A protein from Chlamydophila pneumoniae AR39 had significant similarity compared with protein UbiD from E. coli. Based on this information, the protein-encoding gene was used to swap its counterpart in E. coli, and gene expression in resultant strain DYC was confirmed by RT-PCR. Strain DYC grew using succinate as carbon source and rescued ubiquinone content in vivo, while ubiD deletion strain DYD did not.Results suggest that the chlamydial protein exerts the function of UbiD.

  2. Rac1 regulates the NLRP3 inflammasome which mediates IL-1beta production in Chlamydophila pneumoniae infected human mononuclear cells.

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    Julia Eitel

    Full Text Available Chlamydophila pneumoniae causes acute respiratory tract infections and has been associated with development of asthma and atherosclerosis. The production of IL-1β, a key mediator of acute and chronic inflammation, is regulated on a transcriptional level and additionally on a posttranslational level by inflammasomes. In the present study we show that C. pneumoniae-infected human mononuclear cells produce IL-1β protein depending on an inflammasome consisting of NLRP3, the adapter protein ASC and caspase-1. We further found that the small GTPase Rac1 is activated in C. pneumoniae-infected cells. Importantly, studies with specific inhibitors as well as siRNA show that Rac1 regulates inflammasome activation in C. pneumoniae-infected cells. In conclusion, C. pneumoniae infection of mononuclear cells stimulates IL-1β production dependent on a NLRP3 inflammasome-mediated processing of proIL-1β which is controlled by Rac1.

  3. Isolamento colturale e molecolare di Chlamydophila pneumoniae da pazienti con artropatie. Prospettive patogenetiche e diagnostiche

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    Margherita Giuliodori

    2007-06-01

    Full Text Available Chlamydophila pneumoniae is an ubiquitous intracellular pathogen which causes acute respiratory diseases and may be associated with chronic inflammatory diseases including atherosclerosis, multiple sclerosis and arthritis. C. pneumoniae is rarely cultured from the synovial fluid or blood, and serology is seldom useful. So far most of the studies concerning the possible association between C. pneumoniae and arthritis have been made by molecular methods. Recent advances in the standardization of polymerase chain reaction techniques have shown to confirm a role of C. pneumoniae not only in reactive arthritis (ReA but also in chronic arthritis. In this study, we investigated whether C. pneumoniae could be isolated in synovial fluid and PBMC specimens of patients with different forms of arthritis including ReA.Advanced PCR and Reverse transcriptase PCR techniques targeting different chlamydial genes associated to a novel culture method based on combination of additional centrifugation and extension of culture time, were applied to detect C. pneumoniae in 6 patients with chronic synovitis including one with Anchylosing Spondylitis and relapsing joint swelling. For this patient, serological, coltural as well as molecular assays did detect C. pneumoniae only. Particularly, a high expression of Heat shock protein 60 and 70 of C. pneumoniae was found in the PBMC and the synovial compartments, thus confirming the ability of C. pneumoniae to survive inside blood ad synovia in vital and metabolically active forms. By contrast, the selective decrease of MOMP and 16sRNA, leads to hypotesize a different expression of Chlamydophyla genes during the different phases of infection.

  4. Localizzazione e valutazione dell’espressione di Chlamydophila pneumoniae mediante RT-PCR in situ

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    Stefania Cazzavillan

    2005-12-01

    Full Text Available Chlamydophila pneumoniae, an obligate intracellular gram negative bacterium, is involved in a wide spectrum of symptomatic respiratory tract diseases. However more recently it has been reported to be a pathogenic agent in the mechanism leading to atherosclerosis. In the present study the presence of Chlamydophila pneumoniae was assessed, using nested PCR and in situ PCR, while the viability of the microorganism was investigated using RT in situ PCR.The results obtained demonstrated that Chlamydophila pneumoniae was present and alive in the tissues examined.The global concordance of results in the three techniques used was 100%. RT in situ PCR can be considered a precious tool to detect bacterial mRNA in formalin fixed paraffin embedded samples provided an optimal standardization of the key variables is achieved.

  5. Differences in Cell Activation by Chlamydophila pneumoniae and Chlamydia trachomatis Infection in Human Endothelial Cells

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    Krüll, M.; Kramp, J.; Petrov, T.; Klucken, A. C.; Hocke, A. C.; Walter, C.; Schmeck, B.; Seybold, J.; Maass, M.; Ludwig, S.; Kuipers, Jens G.; Suttorp, N.; Hippenstiel, S.

    2004-01-01

    Seroepidemiological studies and demonstration of viable bacteria in atherosclerotic plaques have linked Chlamydophila pneumoniae infection to the development of chronic vascular lesions and coronary heart disease. In this study, we characterized C. pneumoniae-mediated effects on human endothelial cells and demonstrated enhanced phosphorylation and activation of the endothelial mitogen-activated protein kinase (MAPK) family members extracellular receptor kinase (ERK1/2), p38-MAPK, and c-Jun-NH2 kinase (JNK). Subsequent interleukin-8 (IL-8) expression was dependent on p38-MAPK and ERK1/2 activation as demonstrated by preincubation of endothelial cells with specific inhibitors for the p38-MAPK (SB202190) or ERK (U0126) pathway. Inhibition of either MAPK had almost no effect on intercellular cell adhesion molecule 1 (ICAM-1) expression. While Chlamydia trachomatis was also able to infect endothelial cells, it did not induce the expression of endothelial IL-8 or ICAM-1. These effects were specific for a direct stimulation with viable C. pneumoniae and independent of paracrine release of endothelial cell-derived mediators like platelet-activating factor, NO, prostaglandins, or leukotrienes. Thus, C. pneumoniae triggers an early signal transduction cascade in target cells that could lead to endothelial cell activation, inflammation, and thrombosis, which in turn may result in or promote atherosclerosis. PMID:15501794

  6. Increased infiltration of Chlamydophila pneumoniae in the vessel wall of human veins after perfusion

    NARCIS (Netherlands)

    Kupreishvili, K.; Weeme, M.ter; Morré, S.A.; Brule, A.J.C. van den; Huybregts, M.A.J.M.; Quax, P.H.A.; Velden, J. ten; Hinsbergh, V.W.M. van; Stooker, W.; Eijsman, L.; Niessen, H.W.M.

    2008-01-01

    Background: Several studies have suggested an association between Chlamydophila pneumoniae (Cp) infection and atherosclerosis. A recent study detected Cp DNA in the saphenous vein of 12% of all patients before bypass grafting and in 38% of failed grafts. We used a system in which human veins were pe

  7. Immunoactivity analysis of Cpn 0147 recombinant protein of Chlamydophila pneumoniae%肺炎嗜衣原体Cpn 0147重组蛋白的免疫学活性研究

    Institute of Scientific and Technical Information of China (English)

    代国知; 马忠夏; 周安文; 陈虹亮; 吴移谋

    2012-01-01

    目的 研究Cpn 0147重组蛋白的免疫学活性及其应用于肺炎嗜衣原体(Cpn) 感染临床诊断中的价值.方法 采用GST琼脂糖凝胶纯化目的 蛋白,将Cpn 0147重组蛋白免疫新西兰兔,分别应用Western blot、ELISA法检测其免疫反应性及免疫原性,同时通过检测临床标本以评估其诊断价值.结果 成功表达并纯化了相对分子质量约41×103的重组蛋白GST-Cpn 0147;Western blot和ELISA结果显示,该重组蛋白具有良好的免疫反应性,动物实验结果表明该重组蛋白具有较好的免疫原性,ELISA结果显示该重组蛋白免疫新西兰兔血清抗体效价为1∶12 800.结论 Cpn 0147重组蛋白具有较好免疫学活性和特异性,可为Cpn感染的确诊、预防研究奠定基础.%Objective To explore the immunoactivity of Cpn 0147 recombinant protein,and its clinical value for the diagnosis of Chlamydophila pneumoniae(Cpn) infection. Methods GST-Cpn 014 7 protein was purified by GST sepharose FF. Its immunoactivity was determined by Western blot, and clinical specimens were detected by enzyme linked immunosorbent assay(ELISA). Results The GST fusion protein with molecular weight of 41 × 103 was expressed. The purity of recombination protein after purification reached 95%. Both Western blot and ELISA showed good antigenicity of recombination protein. Conclusion The purified recombination protein might have high specificity and good immunological activity,which could lay a solid foundation for the further study on the serodiagnosis and prevention of Cpn.

  8. The inhibitory effect of disodium cromoglycate on the growth of Chlamydophila (Chlamydia) pneumoniae in vitro.

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    Yamazaki, Tsutomu; Yamaguchi, Tetsuya; Sasaki, Nozomu; Inoue, Miyuki; Sato, Kozue; Kishimoto, Toshio

    2006-04-01

    Chlamydophila (Chlamydia) pneumoniae is associated with asthma and several other respiratory illnesses. Disodium cromoglycate (DSCG) is known to inhibit both immediate and late asthmatic responses. In this study, the inhibitory effect of DSCG on the growth of C. pneumoniae was examined by minimum inhibitory concentration (MIC) and pre-inoculation minimal cidal concentration (MCC) assays using HL cells and C. pneumoniae AR-39. DSCG below the clinically relevant concentration inhibited the growth of C. pneumoniae in a dose-dependent manner in both the MCC and MIC assays. The inhibitory effect was also time-dependent in the MCC assay at 20 mg/ml of DSCG. These results warrant further clinical study on the connection between C. pneumoniae infections and use of DSCG.

  9. Tissue MicroArray (TMA) analysis of normal and persistent Chlamydophila pneumoniae infection.

    Science.gov (United States)

    Borel, Nicole; Mukhopadhyay, Sanghamitra; Kaiser, Carmen; Sullivan, Erin D; Miller, Richard D; Timms, Peter; Summersgill, James T; Ramirez, Julio A; Pospischil, Andreas

    2006-10-19

    Chlamydophila pneumoniae infection has been implicated as a potential risk factor for atherosclerosis, however the mechanism leading to persistent infection and its role in the disease process remains to be elucidated. We validated the use of tissue microarray (TMA) technology, in combination with immunohistochemistry (IHC), to test antibodies (GroEL, GroES, GspD, Ndk and Pyk) raised against differentially expressed proteins under an interferon-gamma (IFN-gamma) induced model of chlamydial persistence. In the cell pellet array, we were able to identify differences in protein expression patterns between untreated and IFN-gamma treated samples. Typical, large chlamydial inclusions could be observed in the untreated samples with all antibodies, whereas the number of inclusions were decreased and were smaller and atypical in shape in the IFN-gamma treated samples. The staining results obtained with the TMA method were generally similar to the changes observed between normal and IFN-gamma persistence using proteomic analysis. Subsequently, it was shown in a second TMA including archival atheromatous heart tissues from 12 patients undergoing heart transplantation, that GroEL, GroES, GspD and Pyk were expressed in atheromatous heart tissue specimens as well, and were detectable morphologically within lesions by IHC. TMA technology proved useful in documenting functional proteomics data with the morphologic distribution of GroEL, GroES, GspD, Ndk and Pyk within formalin-fixed, paraffin-embedded cell pellets and tissues from patients with severe coronary atherosclerosis. The antibodies GroEL and GroES, which were upregulated under persistence in proteomic analysis, displayed positive reaction in atheromatous heart tissue from 10 out of 12 patients. These may be useful markers for the detection of persistent infection in vitro and in vivo.

  10. Tissue MicroArray (TMA analysis of normal and persistent Chlamydophila pneumoniae infection

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    Timms Peter

    2006-10-01

    Full Text Available Abstract Background Chlamydophila pneumoniae infection has been implicated as a potential risk factor for atherosclerosis, however the mechanism leading to persistent infection and its role in the disease process remains to be elucidated. Methods We validated the use of tissue microarray (TMA technology, in combination with immunohistochemistry (IHC, to test antibodies (GroEL, GroES, GspD, Ndk and Pyk raised against differentially expressed proteins under an interferon-gamma (IFN-γ induced model of chlamydial persistence. Results In the cell pellet array, we were able to identify differences in protein expression patterns between untreated and IFN-γ treated samples. Typical, large chlamydial inclusions could be observed in the untreated samples with all antibodies, whereas the number of inclusions were decreased and were smaller and atypical in shape in the IFN-γ treated samples. The staining results obtained with the TMA method were generally similar to the changes observed between normal and IFN-γ persistence using proteomic analysis. Subsequently, it was shown in a second TMA including archival atheromatous heart tissues from 12 patients undergoing heart transplantation, that GroEL, GroES, GspD and Pyk were expressed in atheromatous heart tissue specimens as well, and were detectable morphologically within lesions by IHC. Conclusion TMA technology proved useful in documenting functional proteomics data with the morphologic distribution of GroEL, GroES, GspD, Ndk and Pyk within formalin-fixed, paraffin-embedded cell pellets and tissues from patients with severe coronary atherosclerosis. The antibodies GroEL and GroES, which were upregulated under persistence in proteomic analysis, displayed positive reaction in atheromatous heart tissue from 10 out of 12 patients. These may be useful markers for the detection of persistent infection in vitro and in vivo.

  11. Chlamydophila pneumoniae derived from inclusions late in the infectious cycle induce aponecrosis in human aortic endothelial cells

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    Groscurth Peter

    2008-02-01

    Full Text Available Abstract Background Atherosclerosis is still the leading cause of death in the western world. Besides known risk factors studies demonstrating Chlamydophila pneumoniae (C. pneumoniae to be implicated in the progression of the disease, little is known about C. pneumoniae infection dynamics. We investigated whether C. pneumoniae induce cell death of human aortic endothelial cells, a cell type involved in the initiation of atherosclerosis, and whether chlamydial spots derive from inclusions. Results Lactate dehydrogenase release revealed host cell death to be dependent on the amounts of Chlamydia used for infection. The morphology of lysed human aortic endothelial cells showed DNA strand breaks simultaneously with cell membrane damage exclusively in cells carrying Chlamydia as spots. Further ultrastructural analysis revealed additional organelle dilation, leading to the definition as aponecrotic cell death of endothelial cells. Exclusive staining of the metabolic active pathogens by chlamydial heat shock protein 60 labelling and ceramide incorporation demonstrated that the bacteria responsible for the induction of aponecrosis had resided in former inclusions. Furthermore, a strong pro-inflammatory molecule, high mobility group box protein 1, was shown to be released from aponecrotic host cells. Conclusion From the data it can be concluded that aponecrosis inducing C. pneumoniae stem from inclusions, since metabolically active bacterial spots are strongly associated with aponecrosis late in the infectious cycle in vascular endothelial cells and metabolic activity was exclusively located inside of inclusions in intact cells. Vice versa initial spot-like infection with metabolically inert bacteria does not have an effect on cell death induction. Hence, C. pneumoniae infection can contribute to atherosclerosis by initial endothelial damage.

  12. A novel transport mechanism for MOMP in Chlamydophila pneumoniae and its putative role in immune-therapy.

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    Francis O Atanu

    Full Text Available Major outer membrane proteins (MOMPs of Gram negative bacteria are one of the most intensively studied membrane proteins. MOMPs are essential for maintaining the structural integrity of bacterial outer membranes and in adaptation of parasites to their hosts. There is evidence to suggest a role for purified MOMP from Chlamydophila pneumoniae and corresponding MOMP-derived peptides in immune-modulation, leading to a reduced atherosclerotic phenotype in apoE(-/- mice via a characteristic dampening of MHC class II activity. The work reported herein tests this hypothesis by employing a combination of homology modelling and docking to examine the detailed molecular interactions that may be responsible. A three-dimensional homology model of the C. pneumoniae MOMP was constructed based on the 14 transmembrane β-barrel crystal structure of the fatty acid transporter from Escherichia coli, which provides a plausible transport mechanism for MOMP. Ligand docking experiments were used to provide details of the possible molecular interactions driving the binding of MOMP-derived peptides to MHC class II alleles known to be strongly associated with inflammation. The docking experiments were corroborated by predictions from conventional immuno-informatic algorithms. This work supports further the use of MOMP in C. pneumoniae as a possible vaccine target and the role of MOMP-derived peptides as vaccine candidates for immune-therapy in chronic inflammation that can result in cardiovascular events.

  13. A novel transport mechanism for MOMP in Chlamydophila pneumoniae and its putative role in immune-therapy.

    Science.gov (United States)

    Atanu, Francis O; Oviedo-Orta, Ernesto; Watson, Kimberly A

    2013-01-01

    Major outer membrane proteins (MOMPs) of Gram negative bacteria are one of the most intensively studied membrane proteins. MOMPs are essential for maintaining the structural integrity of bacterial outer membranes and in adaptation of parasites to their hosts. There is evidence to suggest a role for purified MOMP from Chlamydophila pneumoniae and corresponding MOMP-derived peptides in immune-modulation, leading to a reduced atherosclerotic phenotype in apoE(-/-) mice via a characteristic dampening of MHC class II activity. The work reported herein tests this hypothesis by employing a combination of homology modelling and docking to examine the detailed molecular interactions that may be responsible. A three-dimensional homology model of the C. pneumoniae MOMP was constructed based on the 14 transmembrane β-barrel crystal structure of the fatty acid transporter from Escherichia coli, which provides a plausible transport mechanism for MOMP. Ligand docking experiments were used to provide details of the possible molecular interactions driving the binding of MOMP-derived peptides to MHC class II alleles known to be strongly associated with inflammation. The docking experiments were corroborated by predictions from conventional immuno-informatic algorithms. This work supports further the use of MOMP in C. pneumoniae as a possible vaccine target and the role of MOMP-derived peptides as vaccine candidates for immune-therapy in chronic inflammation that can result in cardiovascular events.

  14. Zoonotic atypical pneumonia due to Chlamydophila psittaci: first reported psittacosis case in Taiwan.

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    Cheng, Yu-Jen; Lin, Kun-Yen; Chen, Chun-Chen; Huang, Yen-Lin; Liu, Chun-Eng; Li, Shu-Ying

    2013-07-01

    Human psittacosis caused by Chlamydophila psittaci is one of the most common zoonotic atypical pneumonias featuring pulmonary as well as extrapulmonary infections. Most of the cases involve avian contact history especially with psittacine birds. Herein we report a 44-year-old male patient displaying atypical pneumonia symptoms of intermittent fever, dry cough, chest pain, dyspnea, headache, hepatitis, and hyponatremia. He had two sick cockatiels, one of which had died a month previously. A microimmunofluorescence test was performed to check the serum antibody levels against Chlamydophila psittaci. The serum IgM titer showed positive titer of 1:256, 1:256, and 1:128 on Days 11, 23, and 43 after disease onset, respectively. His fever subsided soon and clinical symptoms improved after minocycline was administrated on Day 12. The psittacosis case was confirmed by history of psittacine bird contact, clinical symptoms, treatment response, and positive IgM titer. To our knowledge, this is the first report of a psittacosis case in Taiwan.

  15. Inhibition of apoptosis in neuronal cells infected with Chlamydophila (Chlamydia pneumoniae

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    Albert Elizabeth V

    2008-01-01

    Full Text Available Abstract Background Chlamydophila (Chlamydia pneumoniae is an intracellular bacterium that has been identified within cells in areas of neuropathology found in Alzheimer disease (AD, including endothelia, glia, and neurons. Depending on the cell type of the host, infection by C. pneumoniae has been shown to influence apoptotic pathways in both pro- and anti-apoptotic fashions. We have hypothesized that persistent chlamydial infection of neurons may be an important mediator of the characteristic neuropathology observed in AD brains. Chronic and/or persistent infection of neuronal cells with C. pneumoniae in the AD brain may affect apoptosis in cells containing chlamydial inclusions. Results SK-N-MC neuroblastoma cells were infected with the respiratory strain of C. pneumoniae, AR39 at an MOI of 1. Following infection, the cells were either untreated or treated with staurosporine and then examined for apoptosis by labeling for nuclear fragmentation, caspase activity, and membrane inversion as indicated by annexin V staining. C. pneumoniae infection was maintained through 10 days post-infection. At 3 and 10 days post-infection, the infected cell cultures appeared to inhibit or were resistant to the apoptotic process when induced by staurosporine. This inhibition was demonstrated quantitatively by nuclear profile counts and caspase 3/7 activity measurements. Conclusion These data suggest that C. pneumoniae can sustain a chronic infection in neuronal cells by interfering with apoptosis, which may contribute to chronic inflammation in the AD brain.

  16. Is there any relationship between Chlamydophila pneumoniae and coronary atherosclerosis among Iranians?

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    Mohammad Hadi Sadeghian

    2013-01-01

    Full Text Available Background: Atherosclerosis is a coronary heart disease, andis the most common cause of death in the industrialized world. Some studies suggested that atherosclerosis may be triggered by infectious agents, mostly Chlamydophila pneumoniae. However, the role of C. pneumoniae in the pathogenesis of coronary atherosclerosis is still controversial. Objectives: This study was performed to evaluate whether there is a significant association between coronary artery atherosclerosis and C. pneumoniae by the polymerase chain reaction (PCR method. Materials and Methods: This case-control study was carried out on formalin-fixed paraffin-embedded tissue biopsies of the coronary arteries obtained from 30 patients with coronary atherosclerosis and 30 subjects without atherosclerosis living in Northeast of Iran. All subjects′ weight and height were determined, and the body mass index was calculated. We also reviewed the medical history and previous laboratory reports of patients. Deoxyribonucleic acid (DNA was extracted, and C. pneumonia DNA was amplified and detected using PCR assay. Results: The age of the patients in the study group was from 18 to 50 years, and the male to female ratio was 5:1. Only oneout of the 30 coronary tissue samples had positive PCR for C. pneumoniae (3.3%, while it was negative for patients in the control group. Conclusions: This study showed that C. pneumoniae infection is not strongly associated with coronary artery atherosclerosis in Northeast of Iran.

  17. Seropositivity ofChlamydophila pneumoniae immunoglobulin G antibody of HIV/AIDS patients in Abuja, Nigeria

    Institute of Scientific and Technical Information of China (English)

    Yakubu Boyi Ngwai; Izebe KS; Ijele IG; Ishaleku D; Inyang US

    2010-01-01

    Objective:To detect IgG antibody toChlamydophila pneumoniae(CP)in sera ofHIV/AIDS patients and provide rationale for inclusion of routine screening for anti-CP antibodies and anti-chlamydial agents in the Nigerian NationalHIV/AIDS Management Plan.Methods: Serum samples from34 consentingHIV/AIDS patients attended a Government-approved Antiretroviral Treatment Facility in Abuja were screened by enzyme-linked immunosorbent assay for anti-CP IgG antibody using ImmunoComb® Chlamydia Bivalent IgG Test kit (Orgenics, Israel).Results:Anti-CP IgG antibody was detected in20 (58.8%)of34 patients tested. The detection rate was higher among the males(8/13; 61.5%) than the females (12/21; 57.1%). Patients of the age group 16-30 years had the highest(7/10; 70%) detection of anti-CP IgG antibody.Conclusions:The result of the present study suggests the presence of anti-CP antibodies in sera of the HIV/AIDS patients, and reinforces the need for routine screening for anti-CPantibodies as a necessary intervention to reduce the burden ofChlamydophila pneumoniae (C. pneumoniae) infections and to reduceHIV-positive morbidity in Nigeria. The outcome of this study also provides justification for the possible inclusion of anti-chlamydial agents in the NationalHIV/AIDS Management Plan to provide prophylaxis against or treat activeC. pneumoniae infections.

  18. Elevata espressione di hsp-60 di Chlamydophila pneumoniae su placche aterosclerotiche carotidee a prognosi infausta

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    Rosario Cultrera

    2007-06-01

    Full Text Available Some difficult microorganisms, including Chlamydophila pneumoniae, are associated with the atherosclerotic tissue damage.The aim of this study was to evaluate the employment of culture together molecular techniques in order to define the possible role of C. pneumoniae in the atherosclerotic tissue damage. Atheromatous carotid plaques (ACP were obtained by endoarterectomies from 10 patients with severe stenosis of the internal carotid artery. Each specimen was divided in three parts: a proximal tract to heart, without stenosis, a medial tract, corresponding to the atheromatous plaque, and a distal tract, above the plaque.Aliquots were employed to perform cultures for C. pneumoniae on Hep - 2 cell line in DMEM. DNA and total RNA were extracted from 50-70 mg. of tissue sample and from Hep - 2 106 cultures to investigate 16S rRNA, momp and hsp60 genes.The PCR and RT-PCR resulted negative for momp gene of C. pneumoniae in all samples. PCR and RT-PCR resulted positive for 16S rRNA or hsp60 genes of C. pneumoniae in the proximal portion of two ACPs with hemorrhagic evolution in two patients, one of which complicated with a retinal tromboembolic outcome. Molecular analyses on C. pneumoniae growing from the cultures are in progress.The DNA and RNA amplification of different portions from ACP seems to be useful to evidence the effective localization of C. pneumoniae in the atheromatous arterial tissue. The highly gene expression of C. pneumoniae hsp60 in a patient with acute hemorrhagic evolution of the carotid plaque may suggest that C. pneumoniae might partecipate in the atherogenesis and induce atherosclerosis complications by inflammatory pathways (activation of cytokines, endothelial factors and matrix-degrading metalloproteinases.

  19. Chlamydophila pneumoniae induces a sustained airway hyperresponsiveness and inflammation in mice

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    Verweij Vivienne

    2007-11-01

    Full Text Available Abstract Background It has been reported that Chlamydophila (C. pneumoniae is involved in the initiation and promotion of asthma and chronic obstructive pulmonary diseases (COPD. Surprisingly, the effect of C. pneumoniae on airway function has never been investigated. Methods In this study, mice were inoculated intranasally with C. pneumoniae (strain AR39 on day 0 and experiments were performed on day 2, 7, 14 and 21. Results We found that from day 7, C. pneumoniae infection causes both a sustained airway hyperresponsiveness and an inflammation. Interferon-γ (IFN-γ and macrophage inflammatory chemokine-2 (MIP-2 levels in bronchoalveolar lavage (BAL-fluid were increased on all experimental days with exception of day 7 where MIP-2 concentrations dropped to control levels. In contrast, tumor necrosis factor-α (TNF-α levels were only increased on day 7. From day 7 to 21 epithelial damage and secretory cell hypertrophy was observed. It is suggested that, the inflammatory cells/mediators, the epithelial damage and secretory cell hypertrophy contribute to initiation of airway hyperresponsiveness. Conclusion Our study demonstrates for the first time that C. pneumoniae infection can modify bronchial responsiveness. This has clinical implications, since additional changes in airway responsiveness and inflammation-status induced by this bacterium may worsen and/or provoke breathlessness in asthma and COPD.

  20. Seroprevalence of Chlamydophila pneumoniae in HIV-infected children in Vietnam.

    Science.gov (United States)

    Trinh, Quang Duy; Pham, Ngan Thi Kim; Le Nguyen, Nhut Tin; Lam, Bao Quoc; Le Phan, Kim Thoa; Truong, Khanh Huu; Le, Thinh Quoc; Nguyen, Hung Thanh; Tang, Thuong Chi; Izumi, Yasuyuki; Komine-Aizawa, Shihoko; Ushijima, Hiroshi; Hayakawa, Satoshi

    2013-06-01

    A total of 89 blood samples collected from HIV-infected infants and children from provinces of southern Vietnam who were hospitalized at Children's Hospital 1, Ho Chi Minh City, during the 1-year period from October 2004 to September 2005 were submitted to serological screening for IgG, IgA, and IgM antibodies against Chlamydophila pneumoniae (C. pneumoniae). The presence of this microorganism was also evaluated by PCR. The results showed that 64 % of the samples were positive for anti-C. pneumoniae IgG, 31.5 % were positive for IgA, and 3.4 % were positive for IgM. The highest prevalences of IgG and IgA positivity, 75 % and 66.7 %, respectively, were noted in the 1- to 2-year-old age group. However, all the samples were negative for C. pneumoniae by PCR. The study revealed a high seroprevalence of C. pneumoniae in Vietnamese infants and children with HIV/AIDS.

  1. The NOD/RIP2 pathway is essential for host defenses against Chlamydophila pneumoniae lung infection.

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    Kenichi Shimada

    2009-04-01

    Full Text Available Here we investigated the role of the Nod/Rip2 pathway in host responses to Chlamydophila pneumoniae-induced pneumonia in mice. Rip2(-/- mice infected with C. pneumoniae exhibited impaired iNOS expression and NO production, and delayed neutrophil recruitment to the lungs. Levels of IL-6 and IFN-gamma levels as well as KC and MIP-2 levels in bronchoalveolar lavage fluid (BALF were significantly decreased in Rip2(-/- mice compared to wild-type (WT mice at day 3. Rip2(-/- mice showed significant delay in bacterial clearance from the lungs and developed more severe and chronic lung inflammation that continued even on day 35 and led to increased mortality, whereas WT mice cleared the bacterial load, recovered from acute pneumonia, and survived. Both Nod1(-/- and Nod2(-/- mice also showed delayed bacterial clearance, suggesting that C. pneumoniae is recognized by both of these intracellular receptors. Bone marrow chimera experiments demonstrated that Rip2 in BM-derived cells rather than non-hematopoietic stromal cells played a key role in host responses in the lungs and clearance of C. pneumoniae. Furthermore, adoptive transfer of WT macrophages intratracheally was able to rescue the bacterial clearance defect in Rip2(-/- mice. These results demonstrate that in addition to the TLR/MyD88 pathway, the Nod/Rip2 signaling pathway also plays a significant role in intracellular recognition, innate immune host responses, and ultimately has a decisive impact on clearance of C. pneumoniae from the lungs and survival of the infectious challenge.

  2. Plasmacytoid dendritic cells prevent cigarette smoke and Chlamydophila pneumoniae-induced Th2 inflammatory responses.

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    Sorrentino, Rosalinda; Gray, Pearl; Chen, Shuang; Shimada, Kenichi; Crother, Timothy R; Arditi, Moshe

    2010-10-01

    Smoking promotes the development of allergic asthma and pneumonia. Chlamydophila pneumoniae lung infection is associated with an increased risk for asthma, inducing an immune response regulated by dendritic cells (DCs). This study sought to determine whether exposure to cigarette smoke modulates the functional activity of CD11c-positive DCs in the lung, with and without concomitant C. pneumoniae infection. Bone marrow-derived DCs (BMDCs) were exposed in vitro to cigarette smoke extract (CSE) and/or live C. pneumoniae (Cpn), and then adoptively transferred intratracheally into wild-type mice. Although CSE plus Cpn appeared to exert an additive effect on the production of Th2 cytokines in vitro, we did not see this effect in vivo. However, the adoptive transfer of DCs pulsed with both CSE and C. pneumoniae into the lungs of naive mice led to an influx of plasmacytoid DCs (pDCs) that suppressed the Th2 skewing ability of the transferred BMDCs. The depletion of pDCs by antibody restored the Th2 skewing ability of the BMDCs. The expression of indoleamine-2,3-dioxygenase in the lung was reduced after the depletion of pDCs, and blocking IFN-α in vitro prevented the ability of pDCs to inhibit the Th2 responses induced by myeloid DCs (mDCs), suggesting their potential involvement in the mechanism of altered polarization. In conclusion, exposure to cigarette smoke skews C. pneumoniae-induced mDCs responses toward a Th2 bias in the lung, which is prevented by pDCs. We propose that pDCs may play a major role in the immunosuppressive lung environment in smokers with C. pneumoniae infection.

  3. The NOD/RIP2 pathway is essential for host defenses against Chlamydophila pneumoniae lung infection.

    Science.gov (United States)

    Shimada, Kenichi; Chen, Shuang; Dempsey, Paul W; Sorrentino, Rosalinda; Alsabeh, Randa; Slepenkin, Anatoly V; Peterson, Ellena; Doherty, Terence M; Underhill, David; Crother, Timothy R; Arditi, Moshe

    2009-04-01

    Here we investigated the role of the Nod/Rip2 pathway in host responses to Chlamydophila pneumoniae-induced pneumonia in mice. Rip2(-/-) mice infected with C. pneumoniae exhibited impaired iNOS expression and NO production, and delayed neutrophil recruitment to the lungs. Levels of IL-6 and IFN-gamma levels as well as KC and MIP-2 levels in bronchoalveolar lavage fluid (BALF) were significantly decreased in Rip2(-/-) mice compared to wild-type (WT) mice at day 3. Rip2(-/-) mice showed significant delay in bacterial clearance from the lungs and developed more severe and chronic lung inflammation that continued even on day 35 and led to increased mortality, whereas WT mice cleared the bacterial load, recovered from acute pneumonia, and survived. Both Nod1(-/-) and Nod2(-/-) mice also showed delayed bacterial clearance, suggesting that C. pneumoniae is recognized by both of these intracellular receptors. Bone marrow chimera experiments demonstrated that Rip2 in BM-derived cells rather than non-hematopoietic stromal cells played a key role in host responses in the lungs and clearance of C. pneumoniae. Furthermore, adoptive transfer of WT macrophages intratracheally was able to rescue the bacterial clearance defect in Rip2(-/-) mice. These results demonstrate that in addition to the TLR/MyD88 pathway, the Nod/Rip2 signaling pathway also plays a significant role in intracellular recognition, innate immune host responses, and ultimately has a decisive impact on clearance of C. pneumoniae from the lungs and survival of the infectious challenge.

  4. Erythema nodosum caused by ascariasis and Chlamydophila pneumoniae pulmonary infection--a case report.

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    Bergler-Czop, Beata; Lis-Swiety, Anna; Kamińska-Winciorek, Grazyna; Brzezińska-Wcisło, Ligia

    2009-12-01

    Erythema nodosum belongs to a group of relatively common hypodermal inflammations. It occurs mainly among women, particularly young women. The etiology of the disease is not clear. Most frequently, changes appear on the surface of the frontal part of the shins. Initially, red nodules change in color to dark brown and then to yellow and green. There is neither dissolution nor cicatrization of the exanthema. Regression is frequent. We present a case of erythema nodosum caused by Ascaris lumbricoides infection as well as by an early Chlamydophila pneumoniae infection, whose etiology has rarely been described in the literature. We were not able to confirm which factor was responsible for the occurrence of the skin changes as treatment of both infections was effective and all skin changes later disappeared completely. Particular attention should be paid to the fact that precise diagnosis of a patient and the search for etiologic factors, even rare ones, are crucial to obtain good results with treatment of erythema nodosum.

  5. Atypical pneumonia

    Science.gov (United States)

    ... people younger than age 40. Pneumonia due to Chlamydophila pneumoniae bacteria occurs year round. Pneumonia due to ... Pneumonia due to mycoplasma and chlamydophila bacteria is usually ... gets worse during the first 4 to 6 days, and then improves over ...

  6. Protein profile of Chlamydophila abortus isolates from Kerala, India

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    Binu K Mani

    Full Text Available Chlamydiae are of microbiological interest because of their mode of interaction with eukaryotic host cells and their specialized life cycle with unique features of parasitism. Reports regarding prevalence of infections of Chlamydophila abortus, the causative organism for chlamydial abortions in livestock, was the basis of the study. Two isolates, one each from cattle and goat abortion along with a reference isolate, were used for characterization with Sodium Dodecyl Sulphate-Poly Acrylamide Gel Electrophoresis (SDS-PAGE. Elementary bodies infected Mc Coy cells, harvested from bottle cultures were disrupted by Teflon coated magnetic pellet. Urografin-76 diluted with Tris-Potassium hydrochloride was used for purification of Elementary bodies of Chlamydophila abortus organism. On protein estimation of Elementary bodies by Biuret method, all the three isolates revealed protein concentration between 500-1000 mg/100ml, which were sufficient for electrophoresis. Ten percent of resolving gel and five percent of stacking gel of polyacrylamide in which 10g of processed isolate samples along with standard protein marker and Mc Coy cell protein (control were electrophoresed. Using Alpha Imager Gel Documentation System, the protein bands were analyzed. Twelve bands each for local bovine isolate and reference isolate were noticed while only 10 bands were there in the caprine isolate. Additional bands of 148 kDa and 135 kDa were present in bovine isolate, compared to the reference isolate, while 152 kDa and 137 kDa bands were unique for caprine isolate. [Vet. World 2011; 4(10.000: 470-472

  7. Fever-triggered Brugada syndrome in an adult patient presenting with hemophagocytic syndrome induced by Chlamydophila pneumoniae.

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    Vieira, Miguel Bigotte; Gaibino, Nuno; Pignatelli, Alexandra; Oliveira, Anabela

    2015-10-09

    A previously healthy 29-year-old man was admitted to our hospital, with a 4-day history of fever (>39°C), rigours, diaphoresis, fatigue and retro-orbital headache. On examination, he was febrile (37.8°C) and tachycardic (110 bpm). Laboratory work up revealed bicytopenia (white cell count 1.37×10(9)/L, platelets 60×10(9)/L) and an increase in C reactive protein (9 mg/dL). The ECG showed ST segment elevation in V1, V2 and V3 leads. The patient was admitted and investigation was initiated revealing prolonged fever (>7 days), pancytopenia, hepatosplenomegaly, hyperferritinemia, hypertriglyceridemia, hypofibrinogenemia, elevated soluble CD25 and hemophagocytosis in bone marrow. Therefore, the patient presented 7 of the 8 diagnostic criteria of hemophagocytic syndrome. Laboratorial investigation for infectious causes was negative, except for IgA and IgG Chlamydophila pneumoniae. ECG re-evaluation on the day of discharge showed no ST segment elevation and no other abnormalities. Genetic testing for known mutations associated with hemophagocytic syndrome and Brugada syndrome did not show any mutations in these genes.

  8. Dendritic Cells from Aged Subjects Display Enhanced Inflammatory Responses to Chlamydophila pneumoniae

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    Sangeetha Prakash

    2014-01-01

    Full Text Available Chlamydophila pneumoniae (CPn is a common respiratory pathogen that causes a chronic and persistent airway infection. The elderly display an increased susceptibility and severity to this infection. However, the underlying mechanisms are not well understood. Dendritic cells (DCs are the initiators and regulators of immune responses. Therefore, we investigated the role of DCs in the age-associated increased CPn infection in vitro in humans. Though the expression of activation markers was comparable between the two age groups, DCs from aged subjects secreted enhanced levels of proinflammatory mediators such as TNF-α and CXCL-10 in response to CPn. In contrast, the secretion of IL-10 and innate interferons, IFN-α and IFN-λ, was severely impaired in DCs from aged donors. The increased activation of DCs from aged subjects to CPn also resulted in enhanced proliferation of CD4 and CD8 T cells in a DC-T coculture. Furthermore, T cells primed with CPn-stimulated DCs from aged subjects secreted increased levels of IFN-γ and reduced levels of IL-10 compared to DCs obtained from young subjects. In summary, DCs from the elderly displayed enhanced inflammatory response to CPn which may result in airway remodeling and increase the susceptibility of the elderly to respiratory diseases such as asthma.

  9. Successful treatment of Chlamydophila pneumoniae acute respiratory distress syndrome with extracorporeal membrane oxygenator: a case report and diagnostic review

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    De Bels David

    2012-01-01

    Full Text Available Abstract Introduction Chlamydophila pneumoniae is a respiratory pathogen known to infect the upper and lower respiratory tracts. Infection severity can range from sub-clinical pulmonary infection to acute respiratory distress syndrome. Case presentation A previously healthy 62-year-old Caucasian man was admitted to our hospital for acute respiratory failure. Serum samples obtained every week starting from the day of admission showed clear-cut seroconversion for C. pneumoniae antibodies. All other cultures obtained during the first days of hospitalization were negative. Despite maximal ventilatory support (high positive end expiratory pressure, fraction of inspired oxygen of 1.0, nitric oxide inhalation, neuromuscular blocking agents and prone positioning, our patient remained severely hypoxemic, which led us to initiate an extracorporeal membrane oxygenation treatment. Extracorporeal membrane oxygenation and hemodiafiltration were withdrawn on day 12. Our patient was extubated on day 18 and discharged from our Intensive Care Unit on day 20. He went home a month later. Conclusion We describe the first published case of acute respiratory distress syndrome due to C. pneumoniae infection successfully treated by extracorporeal membrane oxygenation, a very useful tool in this syndrome. A quick and specific method for the definite diagnosis of Chlamydophila infection should be developed.

  10. 肺炎嗜衣原体热休克蛋白10促炎症作用及其机制的初步研究%Role and preliminary mechanism of heat shock protein 10 of Chlamydophila pneumoniae in inducing inflammatory reaction

    Institute of Scientific and Technical Information of China (English)

    周洲; 陈丽丽; 刘良专; 李忠玉; 游晓星; 陈曦; 吴移谋

    2013-01-01

    目的 探讨肺炎嗜衣原体(Chlamydophila pne umoniae,Cpn)热休克蛋白(heat shock protein,HSP)10(CHSP10)诱导人单核细胞分泌炎症因子的作用及Toll样受体(Toll-like receptor,TLR)2、TLR4与此作用的相关性.方法 以去内毒素活性的不同质量浓度CHSP10(0.5、1、5、10、20、30 μg/ml)刺激THP-1细胞0、6、12、24、36、48、60 h,检测蛋白未处理组、加热处理组、去蛋白处理组中IL-1β及IL-6水平;用CHSP10刺激C3H系野生型(C3H/HeN)和TLR4缺陷型(C3H/HeJ)小鼠腹腔巨噬细胞,分别检测II-1β及IL-6水平;用CHSP10刺激被抗TLR2/TLR4抗体处理的THP-1细胞,检测IL-1β、IL-6的变化.结果 CHSP10可诱导THP-1细胞产生炎症因子IL-1β、IL-6;CHSP10诱生C3H系野生型小鼠细胞分泌的炎症因子明显高于TLR4缺陷型小鼠细胞;用TLR2和/或TLR4抗体封闭后,CHSP10诱生的IL-1β、IL-6不同程度减少.结论 CHSP10可能作为炎症相关蛋白参与了Cpn对宿主细胞的致炎作用;并且TLR2及TLR4在该炎症刺激信号的传递过程中发挥一定的作用.%To investigate the role and mechanism of heat shock protein 10 (HSP10) of Chlamydophila pneumoniae in inducing inflammatory reaction. Purified native recombinant HSP10 from C. pneumoniae (CHSP10) were inactivated for cleaning contaminated endotoxin and were used to stimulate THP -1 with different concentrations for different time, then the cytokines levels were measured by using human enzyme -linked immunosorbent assay kit. Peritoneal macrophages from TLR4-deficient mice and wide-type mice were stimulated with endotoxin -free proteins respectively, and the cytokines levels were measured. Furthermore, blocking experiments with neutralizing anti-human TLR2 or/and TLR4 MAb were carried out and ELISA was used to detect the concentration of cytokines. Result showed that HSP10 was able to induce IL—1β and IL-6 secretion in THP-1. Wild-type macrophages from C3H/HeN secreted higher IL—1β and IL-6 level

  11. Effect of the termini of RNase Hs from Chlamydophila pneumoniae on enzymatic biochemical characterization

    Institute of Scientific and Technical Information of China (English)

    Jingli Hou; Zheng Lu; Xingliang Guo; Jianhua Liu

    2012-01-01

    A difference between prokaryotic RNase HⅡ and HⅢ,which both belong to type 2 RNase H,is a long N-terminal extension of HⅢ; however,the main-fold structures of HⅡ and HⅢ known as RNase H-fold are similar.To further understand the structure-function relationship of RNase HⅡ and RNase HⅢ,biochemical analyses were carried out using N-terminal truncations of RNase HⅢ (ⅢN56Δ,ⅢN81Δ,and ⅢN88Δ) and C-terminal truncation (ⅡC19Δ) of RNase HⅡ from Chlamydophila pneumoniae.Compared with wild-type CpRNase HⅡ/Ⅲ,IIIN56Δhad no obvious variation on the cleavage site and efficiency of DNA-rN1-DNA/DNA (DR1D) and DNA-rN4-DNA/DNA (DR4D) substrates.ⅡC19Δ and ⅢN81Δ both showed decreased activities,and ⅢN88Δ exhibited little cleavage on these substrates.However,ⅢN81Δ showed very different activities toward different substrates (20%for DR1D and 85% for DR4D).Moreover,ⅡC19ΔⅢN82-88 mutant,prepared through adding N-terminal 82nd to 88th residues locating at the bound region of N-and C-terminal domains of CpRNase HⅢ to N-terminus of ⅡC19Δ,cleaved DR4D substrate more efficiently and preferentially at the cleavage sites of CpRNase HⅢ but not those of CpRNase HⅡ.These results indicated that C-termini of CpRNase HⅡ,N-termini of CpRNase HⅢ,and bound region of N-and C-terminal domain are all important for enzymatic activities.Moreover,the 82nd to 88th residues of N-terminus of CpRNase HⅡ are related with enzyme cleavage site specificity.These results will help to understand the importance of C-termini of CpRNase HⅡ and N-termini of CpRNase HⅢ to the enzyme activities for DR1D and DR4D substrate.

  12. The validity of the criteria for primary infection of Chlamydophila pneumoniae in children by measuring ELISA IgM antibodies.

    Science.gov (United States)

    Kamata, Ayako; Obinata, Kaoru; Niizuma, Takahiro; Kinoshita, Keiji; Shimizu, Toshiaki

    2012-06-01

    As IgM antibody measurement by enzyme-linked immunosorbent assay (ELISA) has become possible for the serological diagnosis of Chlamydophila pneumoniae (C. pn) infection, the HITAZYME-ELISA method has become widely employed in Japan. However, in children, when the diagnostic criterion of primary infection is set at ID ≥1.1, the positive rate is higher than expected, and the potential for inaccurate reflection of the prevalence has been raised. In this study, we performed ROC analysis involving 136 pediatric patients with acute airway symptoms (0-14 years of age), considering a 32-fold or higher micro-immunofluorescence IgM antibody titer against C. pn as positive. Setting the cut-off value for ELISA C. pn IgM antibody ID at 2.0, the specificity was 100%, with no false positivity. The maximum (sensitivity + specificity)/2 was obtained when the cut-off value was set at 1.5. Therefore, IgM ID ≥2.0 was regarded as definitely positive and an IgM ID between 1.5 and 2.0 was regarded as indeterminate as diagnostic criteria for the primary infection. When the prevalence was investigated in 3,108 children (0-15 years of age) with airway symptoms based on these criteria, 542 cases (17.4%) were positive, and the median duration of IgM antibody positivity was five months. Long-term positivity (ten cases) for more than 12 months and recurrent positivity (eight cases) were also observed, but it may be appropriate to set a new criterion of IgM antibody ID ≥2.0 for the diagnosis of primary Chlamydophila pneumoniae infection in children.

  13. Cloning and expression of Cpn0425 recombinant protein from Chlamydophila pneumoniae and its antigenicity%肺炎嗜衣原体Cpn0425重组蛋白的克隆与表达及抗原性研究

    Institute of Scientific and Technical Information of China (English)

    肖光文; 刘良专; 徐磊; 谢小平; 吴移谋

    2012-01-01

    The aim of this study is to construct the rccombinant protein of Cpn0425 from Chlamydop hila pneumoniae and induce its expression in E. coli, meanwhile assess the antigenicity of protein Cpn0425 . The gene of Cpn0425 was amplified by PCR , which was used to construct the recombinant plasm id of pGEX6p-2/Cpn0425 , and then the products were identified and analyzed by SDS-PAGE and Western blot . The restriction enzymes cleavage analysis and nncleotide sequencing showed the target gene was successfully inserted into pGEX 6p-2 prokaryotic expression vector . The similarity was 100% between the in-serted gene and Cpn0425 gene reported in GenBank by BLAST analysis . The SDS-PAGE demonstrated that the GST-Cpn0425 recombinant protein with a relative molecular weight about 50 kDa was expressed after the E. coli BL21 contained recombinant plasmid was induced and mainly existed in the pattern of solubility . Its purity reached up to 95% after purification with gluta-thione S-transferase (GST) resin chromatography of Novagen . In conclusion , we obtain Cpn0425 gene of Gt and express it suc-cessfully in E. coli BL21 . The GST-Cpn0425 has a good antigenicity.%目的 构建重组质粒pGEX6p-2/Cpn0425,在E.coli BL21中进行诱导表达,纯化获得重组融合蛋白Cpn0425,并评价其抗原性.方法 PCR扩增肺炎嗜衣原体Cpn0425蛋白编码基因,构建pGEX6p-2/Cpn0425重组质粒,测序正确后在E.coli BL21中诱导表达,用SDS-PAGE和 Western-Blot 进行分析和鉴定.结果 构建了重组质粒pGEX6p-2/Cpn0425,并在E.coli BL21菌中高效表达出Mr约为50kDa的GST-Cpn0425目的 蛋白,超声裂解后SDS-PAGE分析目的 蛋白在菌体细胞内以可溶性形式表达,经采用默克Novagen的GST纯化树脂纯化获得纯度在95%以上的重组蛋白.结论 获得了Cpn0425基因片段,并在E.coli BL21中成功表达,GST-Cpn0425具有良好的抗原性.

  14. Study on the inflammatory injury of lung tissue and influence on inflammatory cytokines in mouse induced by recombinant protein CPAF from Chlamydophila pneumoniae%肺炎嗜衣原体CPAF重组蛋白致小鼠肺组织炎症及对炎症细胞因子的影响

    Institute of Scientific and Technical Information of China (English)

    王华丽; 吴移谋; 郑江花; 朱翠明; 李忠玉; 周洲; 余敏君

    2011-01-01

    目的:研究肺炎嗜衣原体(Chlamydophila pneumoniae,Cpn) 蛋白酶样活性因子(CPAF)的181~400aa基因(CPAFm)的重组蛋白在BALB/c小鼠体内引起的肺部炎症改变及对炎症细胞因子TNF-α和IL-6的水平的影响,为进一步探索CPAF在Cpn感染中的致病作用提供实验依据.方法:将纯化的CPAFm重组蛋白于鼻内或尾静脉注射BALB/c小鼠, HE染色观察肺组织病理形态学改变,计小鼠外周血与支气管肺泡灌洗液(Bronchoalveolar lavage fluids,BALF)中白细胞总数,ELISA方法检测血清和BALF中的TNF-α和IL-6的水平.结果:实验组BALB/c小鼠的肺组织出现中性粒细胞、淋巴细胞等炎症细胞浸润,对照组与正常组BALB/c小鼠肺组织未见明显病理改变.鼻内滴入重组蛋白实验组BALB/c小鼠BALF中白细胞总数明显高于GST对照组(P<0.05)、PBS对照组(P<0.05)和正常组(P<0.01);鼻内滴入和尾静脉注射重组蛋白实验组BALF中炎症因子IL-6、TNF-α的水平明显高于GST、PBS对照组与正常组(均P<0.01);鼻内滴入重组蛋白实验组外周血中炎症因子IL-6和TNF-α水平显著高于GST、PBS对照组和正常组(P<0.05);尾静脉注射重组蛋白实验组外周血中炎症因子IL-6的水平明显高于相应的对照组(均P<0.01).结论:CPAFm重组蛋白有致病性,能引起BALB/c小鼠肺组织的炎症损伤和炎症因子TNF-α、IL-6的水平的升高,肺损伤与BALB/c小鼠体内炎症细胞增多和炎症因子TNF-α、IL-6的水平的升高相关.%Objective :To provide experimental basis for exploring pathogenic mechanism of Chlamydophila pneumoniae ,and to investigate the changes of pulmonary inflammation and inflammatory cytokines inclucling TNF-α and IL-6 levels in mouse model .Methods :The gene fragment of Chlamydial protease-like activity factor(CPAF)from C .pneumoniae (CPAFm ) was cloned , which encoded 181~400aa of the protein .The gene fragment was inserted into plasmid (name) and expressed in

  15. Bioinformatic analysis of Chlamydophila pneumoniae Cpn0797 protein and the preparation of its monoclonal antibody%肺炎嗜衣原体Cpn0797蛋白的生物信息学分析及其单克隆抗体制备鉴定

    Institute of Scientific and Technical Information of China (English)

    陈超群; 吴移谋; 任林; 刘良专

    2012-01-01

    目的 生物信息学分析肺炎嗜衣原体Cpn0797蛋白的结构,制备特异性的抗Cpn0797蛋白的单克隆抗体.方法 用ProtParam、SignalP、NPS@、和PSORT等软件对Cpn0797蛋白的理化参数、信号肽、二级结构、蛋白亚细胞定位进行分析;原核表达纯化GST-Cpn0797融合蛋白,以其为免疫原免疫BALB/c小鼠.杂交瘤技术制备单克隆抗体,采用间接免疫荧光法鉴定单克隆抗体的亚类和特异性.结果 生物信息学分析表明Cpn0797蛋白二级结构以无规则卷曲为主;成功地建立了能稳定分泌抗Cpn0797蛋白的单克隆抗体杂交瘤细胞株,单克隆抗体能特异性的识别Cpn0797内源性蛋白.结论成功制备特异性的Cpn0797单克隆抗体,为进一步探究Cpn0797蛋白的生物学功能提供实验基础.%In order to analyze the protein structure of Chlamydophila pneumoniae Cpn0797 protein and to prepare spe cific monoclonal antibodies (mAb) against Cpn0797 protein, ProtParam, SignalP, NPS@ and PSORT software packages were used to predict the physical and chemical properties, signal peptide, secondary structure and protein localization sites in cells according to the amino acid sequence of Cpn0797. The gene cpn0797 was cloned into the prokaryotic expression vector pGEX6p 2, and expressed in E. Coli and then purified. The mAbs against Cpn0797 were prepared with the hybridoma tech nique after mice were immunized with purified GST Cpn0797 fusion protein. The isotype and specificities of the mAbs were de termined by indirect immunofluorescence assay (IFA). The bioinformatical analysis results showed that Cpn0797 protein main ly consisted of random coils. Hybridoma cell lines stably secreting mAbs against Cpn0797 protein are obtained. The mAbs re acted with Cpn0797 endogenous protein. In conclusion, the specific mAbs against Cpn0797 protein were obtained, which pro vides a basis for further study in the biological function of Cpn0797 protein.

  16. Detection of IgM and IgG antibodies to Chlamydophila pneumoniae in pediatric community-acquired lower respiratory tract infections

    Directory of Open Access Journals (Sweden)

    Surinder Kumar

    2011-01-01

    Full Text Available Context: Chlamydophila pneumoniae (C. pneumoniae is an emerging infectious agent with a spectrum of clinical manifestations including lower and upper respiratory tract infections. Aims: To investigate the role of C. pneumoniae in community-acquired lower respiratory tract infections (LRTIs in children using serological tests. Settings and Design: Two hundred children, age 2 months to 12 years, hospitalized for community-acquired LRTIs were investigated for C. pneumoniae etiology. Materials and Methods: We investigated 200 children hospitalized for community-acquired LRTIs, using ELISA for detecting anti-C. pneumoniae IgM and IgG antibodies. The demographic, clinical and radiological findings for C. pneumoniae antibody positive and C. pneumoniae antibody negative cases were compared. Statistical Analysis Used: Data analysis was performed by Chi-square test and Fisher′s exact tests using Epi Info (2002. Results: Clinical and radiological findings in both the groups were comparable. Serological evidence of C. pneumoniae infection was observed in 12 (6% patients; specific IgM antibodies were detected in 11 (91.67%; specific IgG antibodies in 1 (8.33% patients, while 4-fold rise in C. pneumoniae IgG antibody titers were noted in none of the patients. Conclusions: C. pneumoniae has a role in community-acquired LRTIs, even in children aged < 5 years. Serological detection using ELISA would enable pediatricians in better management of C. pneumoniae infections.

  17. Identification, sequencing and molecular analysis of Chp4, a novel chlamydiaphage of Chlamydophila abortus belonging to the family Microviridae.

    Science.gov (United States)

    Sait, Michelle; Livingstone, Morag; Graham, Rebecca; Inglis, Neil F; Wheelhouse, Nick; Longbottom, David

    2011-07-01

    Members of the family Microviridae have been identified in a number of chlamydial species infecting humans (phage CPAR39 in Chlamydophila pneumoniae), other mammals (φCPG1 in Chlamydophila caviae, Chp2 in Chlamydophila abortus and Chp3 in Chlamydophila pecorum) and birds (Chp1 in Chlamydophila psittaci). This study describes the identification and genome sequencing of Chp4, an icosahedral, 4530 bp, ssDNA phage in C. abortus. Chp4 is predicted to contain eight ORFs, six of which could be assigned putative functions based on sequence similarity to characterized bacteriophage. Gene order and content were highly conserved amongst chlamydiaphage, with the highest sequence variability occurring in the IN5 and INS variable regions of the VP1 major coat protein, which has been associated with host cell recognition and binding. Phylogenetic analysis of VP1 indicated that Chp4 is a member of the Chlamydiamicrovirus, and is most closely related to phage φCPG1 and CPAR39.

  18. Application of serology and nested polymerase chain reaction for identifying Chlamydophila pneumoniae in community-acquired lower respiratory tract infections in children

    Directory of Open Access Journals (Sweden)

    Surinder Kumar

    2016-01-01

    Full Text Available Context: Chlamydophila pneumoniae is a common cause of community-acquired respiratory infections, including pneumonia, bronchitis, and upper respiratory tract infections. Since it is difficult to detect C. pneumoniae in clinical practice, specific etiological diagnosis is established only in a minority of cases. Aims: To investigate the role of C. pneumoniae in community-acquired lower respiratory tract infections (LRTIs in children, with the use of serological tests and nested polymerase chain reaction (PCR analysis. Settings and Design: One hundred children, age of 2 months to 12 years, hospitalized for community-acquired LRTIs were investigated for C. pneumoniae etiology. Materials and Methods: We investigated 100 children hospitalized for community-acquired LRTIs, using enzyme-linked immunosorbent assay for detecting anti-C. pneumoniae immunoglobulin M, and immunoglobulin G antibodies and nasopharyngeal aspirates for analysis of C. pneumoniae PCR. The demographic, clinical, and radiological findings for C. pneumoniae antibody positive and C. pneumoniae antibody negative cases were compared. Statistical Analysis Used: Data analysis was performed by Chi-square test and Fisher′s exact tests using Epi Info (2002. Results: Clinical and radiological findings in both the groups were comparable. A relatively higher rate of C. pneumoniae infection in children was observed below 5 years of age. Serological evidence of C. pneumoniae infection was observed in 12 (12% patients and nested PCR was positive in 5 (5% children. Thirteen (13% patients were diagnosed with C. pneumoniae infection by serology and/or nested PCR. Conclusions: Our study confirms that C. pneumoniae plays a significant role in community-acquired LRTIs in children of all ages, even in children aged <5 years.

  19. Secretion of TNF-α induced by heat shock protein 10 of Chlamydophila pneumoniae were mediated by TLR2 and TLR4 in THP-1%肺炎嗜衣原体热休克蛋白10经TLR2及TLR4调控THP-1分泌TNF-α

    Institute of Scientific and Technical Information of China (English)

    周洲; 杨科; 陈丽丽; 陈虹亮; 李忠玉; 吴移谋

    2012-01-01

    目的 探讨肺炎嗜衣原体(Cpn)热休克蛋白10(HSP10)诱导人单核细胞分泌炎症因子的作用及Toll样受体(TLR)2、TLR4与此作用的相关性.方法 制备Cpn HSP10(CHSP10)纯化蛋白,去内毒素活性后用不同浓度(0.5、1、5、10、20、30μ/ml)刺激THP-1细胞0、6、12、24、36、48、60 h,并比较蛋白不同处理组别中TNF-α的水平差异;以间接免疫荧光及RT-PCR鉴定THP-1细胞上的TLR4及TLR2;分离C3H系野生型和TLR4缺陷型小鼠巨噬细胞,以CHSP10刺激后检测TNF-α水平;用抗TLR2/TLR4单克隆抗体预孵育细胞,ELISA检测CHSP10刺激细胞前后TNF-α的变化.结果 CHSP10刺激THP-1细胞引起上清液炎症因子TNF-α水平显著增加,经加热等处理后蛋白诱生TNF-α的作用明显降低.THP-1细胞可检测到TLR2及TLR4的mRNA及蛋白表达,CHSP10诱生C3H系野生型小鼠细胞分泌的TNF-α明显高于TLR4缺陷型小鼠细胞,TLR2/4经单克隆抗体作用后均可显著降低CHSP10诱导THP-1分泌TNF-α的水平.结论 CHSP10可能作为炎症相关蛋白参与了Cpn对宿主细胞的致炎作用;并且TLR2及TLR4在该炎症刺激信号的传递过程中发挥一定的作用.%Objective To investigate the effect of heat shock protein 10 (HSP1O) of Chlamydophila pneumoniae in inducing TNF-α on THP-1 cells and the roles of TLR4 and TLR2 involved in it.Methods Purified native recombinant HSP10 from Cpn(CHSP10) were produced and inactivated the endotoxin contamination,then different concentration (0.5,1,5,10,20,30 μg/ml) of CHSP10 were used to stimulate THP-1 for different time (0,6,12,24,36,48,60 h).TNF-α were measured by using human TNF-α ELISA kit and compared among different groups.THP-1 were collected and analyzed for TLR2 and TLR4 mRNA levels and protein expression by RT-PCR and immunofluorescence.Peritoneal macrophages isolated from wide-type (C3 H/HeN) and TLR4-deficient mice (C3H/HeJ) were stimulated with endotoxin-free proteins respectively,and the TNF

  20. Chlamydophila felis CF0218 is a novel TMH family protein with potential as a diagnostic antigen for diagnosis of C. felis infection.

    Science.gov (United States)

    Ohya, Kenji; Takahara, Yu; Kuroda, Etsuko; Koyasu, Saori; Hagiwara, Shigeyuki; Sakamoto, Maki; Hisaka, Mitsuaki; Morizane, Kazuko; Ishiguro, Shinryou; Yamaguchi, Tsuyoshi; Fukushi, Hideto

    2008-10-01

    Chlamydophila felis is a causative agent of acute and chronic conjunctivitis and pneumonia in cats (feline chlamydiosis). Also, C. felis is a suspected zoonotic agent of such diseases as non-Chlamydia trachomatis conjunctivitis in humans, although this is controversial. At present, there is no serodiagnostic system that specifically detects C. felis infection conveniently. Current systems use antigens such as lipopolysaccharide that cross-react with all chlamydia species. In addition, it is difficult to distinguish between cats that are vaccinated with the commercial vaccine against C. felis and cats that are infected with C. felis. Here, we describe a new candidate diagnostic antigen for diagnosis of C. felis infection, CF0218, that was obtained by screening a genomic expression library of C. felis Fe/C-56 with C. felis-immunized serum. CF0218 was a putative transmembrane head (TMH) family protein with bilobed hydrophobic motifs at its N terminus, and orthologues of CF0218 were not found in the Chlamydophila pneumoniae or Chlamydia trachomatis genomes. The recombinant CF0218 was not recognized by antiserum against C. trachomatis, suggesting that CF0218 is C. felis specific. CF0218 transcription during the course of C. felis infection was confirmed by reverse transcription-PCR. By indirect immunofluorescence analysis, CF0218 was colocalized with the C. felis-formed inclusion bodies in the infected cells. The antibody response against CF0218 was elevated following C. felis infection but not by vaccination in experimentally vaccinated and infected cats. These results suggest that CF0218, a novel TMH family protein of C. felis, possesses potential as a C. felis infection-specific diagnostic antigen.

  1. Ateroesclerosis asociada a infección por chlamydophila pneumoniae: interacción entre el ser humano y una bacteria

    Directory of Open Access Journals (Sweden)

    Alvaro Gudiño-Gomezjurado

    2015-04-01

    Full Text Available La ateroesclerosis y sus complicaciones constituyen una de las mayores causas de morbi-mortalidad a nivel mundial. Sin embargo, solo la mitad de los casos se justifica por los factores de riesgo tradicionales. No obstante, desde hace casi cinco décadas se ha intentado encontrar causas no tradicionales asociadas a la enfermedad ateroesclerótica. Varios agentes infecciosos han emergido como posibles candidatos centralizándose la investigación en Chlamydophila pneumoniae por su capacidad de infectar las células endoteliales durante el proceso ateroesclerótico. A lo largo del tiempo, múltiples estudios han tratado de probar la causalidad de C. pneumoniae en el proceso ateroesclerótico y sus complicaciones. A pesar de esto, hasta la fecha las conclusiones son ambiguas y de poca relevancia para la práctica clínica diaria.

  2. Polymorphic membrane proteins 1 and 7 from Chlamydophila felis are significant immunodominant proteins.

    Science.gov (United States)

    Harley, Ross; Catanese, Bernardo; Helps, Chris

    2010-08-26

    Chlamydophila felis is a common cause of conjunctivitis in cats. Greater understanding of C. felis infection and immunity and identification of protective antigens will facilitate improved vaccine design. Chlamydial polymorphic membrane proteins (Pmps) represent a family of homologous proteins of likely importance in chlamydial infection and immunity. To identify immunogenic C. felis Pmps, we generated recombinant C. felis Pmps (rPmps) and used these to detect serum antibody reactivity against Pmps arising during C. felis infection in cats. Sequencing of Pmp genes 1, 7, 13, 18, 19 and 20 from 3 laboratory strains of C. felis (K2487, 1497V and Cello) and alignment with the Fe/C-56 genome revealed high genetic identity in Pmp genes between strains. PCR products lacking the predicted N-terminal signal sequence peptide and C-terminal domain were generated, cloned and expressed in Escherichia coli prior to purification by nickel-agarose affinity chromatography. Serum samples from 4 cats collected up to 55 days post-inoculation with C. felis (K2487) were analysed by western blotting and rPmp-specific ELISAs for evidence of serum antibody reactivity against each rPmp. Strong serum antibody reactivity against rPmps 1 and 7, and weak heterogeneous serum immunoreactivity against rPmps 13, 19 and 20, were detected from 14 to 21 days post-infection (dpi), peaking at 28-35 dpi and tending to plateau thereafter. No significant serum antibody reactivity was detected against rPmp18. This study provides the first evidence that C. felis Pmps 1 and 7 are likely to represent immunodominant proteins and recommends investigation of their potential as serodiagnostic antigens and novel vaccine candidates.

  3. Evaluation and optimization of a commercial enzyme linked immunosorbent assay for detection of Chlamydophila pneumoniae IgA antibodies

    Directory of Open Access Journals (Sweden)

    Gargouri Jalel

    2008-07-01

    Full Text Available Abstract Background Serologic diagnosis of Chlamydophila pneumoniae (Cpn infection routinely involves assays for the presence of IgG and IgM antibodies to Cpn. Although IgA antibodies to Cpn have been found to be of interest in the diagnosis of chronic infections, their significance in serological diagnosis remains unclear. The microimmunofluorescence (MIF test is the current method for the measurement of Cpn antibodies. While commercial enzyme linked immunosorbent assays (ELISA have been developed, they have not been fully validated. We therefore evaluated and optimized a commercial ELISA kit, the SeroCP IgA test, for the detection of Cpn IgA antibodies. Methods Serum samples from 94 patients with anti-Cpn IgG titers ≥ 256 (study group and from 100 healthy blood donors (control group were tested for the presence of IgA antibodies to Cpn, using our in-house MIF test and the SeroCP IgA test. Two graph receiver operating characteristic (TG-ROC curves were created to optimize the cut off given by the manufacturer. Results The MIF and SeroCP IgA tests detected Cpn IgA antibodies in 72% and 89%, respectively, of sera from the study group, and in 9% and 35%, respectively, of sera from the control group. Using the MIF test as the reference method and the cut-off value of the ELISA test specified by the manufacturer for seropositivity and negativity, the two tests correlated in 76% of the samples, with an agreement of Ƙ = 0.54. When we applied the optimized cut-off value using TG-ROC analysis, 1.65, we observed better concordance (86% and agreement (0.72 between the MIF and SeroCP IgA tests. Conclusion Use of TG-ROC analysis may help standardize and optimize ELISAs, which are simpler, more objective and less time consuming than the MIF test. Standardization and optimization of commercial ELISA kits may result in better performance.

  4. The relationship of Chlamydophila pneumoniae with schizophrenia: The role of brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3) in this relationship.

    Science.gov (United States)

    Kalayci, Fatma; Ozdemir, Armagan; Saribas, Suat; Yuksel, Pelin; Ergin, Sevgi; Kuskucu, Ali Mert; Poyraz, Cana Aksoy; Balcioglu, Ibrahim; Alpay, Nihat; Kurt, Aykut; Sezgin, Zeynep; Kocak, Banu Tufan; Icel, Rana Sucu; Can, Gunay; Tokman, Hrisi Bahar; Kocazeybek, Bekir

    Several pathogens have been suspected of playing a role in the pathogenesis of schizophrenia. Chronic inflammation has been proposed to occur as a result of persistent infection caused by Chlamydophila pneumoniae cells that reside in brain endothelial cells for many years. It was recently hypothesized that brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3) may play prominent roles in the development of schizophrenia. NT-3 and BDNF levels have been suggested to change in response to various manifestations of infection. Therefore, we aimed to elucidate the roles of BDNF and NT3 in the schizophrenia-C. pneumoniae infection relationship. RT-PCR, immunofluorescence and ELISA methods were used. Fifty patients suffering from schizophrenia and 35 healthy individuals were included as the patient group (PG) and the healthy control group (HCG), respectively. We detected persistent infection in 14 of the 50 individuals in the PG and in 1 of the 35 individuals in the HCG. A significant difference was found between the two groups (p0.05). C. pneumoniae DNA was not detected in any group. A significant difference in NT-3 levels was observed between the groups, with very low levels in the PG (p0.05). In conclusion, we suggest that NT-3 levels during persistent C. pneumoniae infection may play a role in this relationship. Copyright © 2016 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  5. Chlamydophila pneumoniae HflX belongs to an uncharacterized family of conserved GTPases and associates with the Escherichia coli 50S large ribosomal subunit.

    Science.gov (United States)

    Polkinghorne, Adam; Ziegler, Urs; González-Hernández, Yanela; Pospischil, Andreas; Timms, Peter; Vaughan, Lloyd

    2008-11-01

    Predicted members of the HflX subfamily of phosphate-binding-loop guanosine triphosphatases (GTPases) are widely distributed in the bacterial kingdom but remain virtually uncharacterized. In an attempt to understand mechanisms used for regulation of growth and development in the chlamydiae, obligate intracellular and developmentally complex bacteria, we have begun investigations into chlamydial GTPases; we report here what appears to be the first analysis of a HflX family GTPase using a predicted homologue from Chlamydophila pneumoniae. In agreement with phylogenetic predictions for members of this GTPase family, purified recombinant Cp. pneumoniae HflX was specific for guanine nucleotides and exhibited a slow intrinsic GTPase activity when incubated with [gamma-(32)P]GTP. Using HflX-specific monoclonal antibodies, HflX could be detected by Western blotting and high-resolution confocal microscopy throughout the vegetative growth cycle of Cp. pneumoniae and, at early time points, appeared to partly localize to the membrane. Ectopic expression of Cp. pneumoniae HflX in Escherichia coli revealed co-sedimentation of HflX with the E. coli 50S large ribosomal subunit. The results of this work open up some intriguing possibilities for the role of GTPases belonging to this previously uncharacterized family of bacterial GTPases. Ribosome association is a feature shared by other important conserved GTPase families and more detailed investigations will be required to delineate the role of HflX in bacterial ribosome function.

  6. Early diagnosis using recombinant protein of immunodominant region gene of chlamydial protease-like activity factor from Chlamydophila pneumoniae%肺炎嗜衣原体蛋白酶样活性因子免疫优势区基因重组蛋白在早期诊断中的应用

    Institute of Scientific and Technical Information of China (English)

    郑江花; 吴移谋; 刘佳强; 刘广超; 陈超群

    2008-01-01

    [目的]克隆和表达肺炎嗜衣原体(Chlamydophila pneumoniae,Cpn)蛋白酶样活性因子(CPAF)免疫优势区基因,评价重组蛋白在早期感染诊断中的应用价值.[方法]挑选并克隆出Cpn CPAF免疫优势区基因,构建原核表达载体,诱导表达并纯化重组蛋白,分析其抗原特异性;间接ELISA法检测Cpn参考血清、临床血清标本中的特异性IgM抗体,以及呼吸道感染患者痰咽拭子中的Cpn抗原;检测沙眼衣原体(Chlamydia trachomatis,Ct)临床阳性血清和泌尿生殖道分泌物.[结果]高效表达和纯化出一相对分子量约51.3kDa的重组蛋白;Western blot证明其只与人抗Cpn抗血清发生特异性反应;间接ELISA法检测40份Cpn IgM参考血清,阴性和阳性结果的一致率均为100%(40/40);与"金标准"方法MIF对照,检测300例临床血清标本中的IgM抗体,符合率为98.3%;与PCR试剂对照,检测120份呼吸道感染患者痰咽拭子中的Cpn抗原,符合率为88.3%;检测Ct阳性血清和泌尿生殖道分泌物,与Ct没有交叉反应.[结论]制备的CPAF免疫优势区基因重组蛋白具有良好的抗原性,在Cpn感染早期诊断中具有较高的利用价值.

  7. Identification and characterization of Inc766, an inclusion membrane protein in Chlamydophila abortus-infected cells.

    Science.gov (United States)

    Vretou, Evangelia; Katsiki, Evangelia; Psarrou, Evgenia; Vougas, Kostantinos; Tsangaris, George Th

    2008-10-01

    We have identified the gene product of locus 766 in the transmembrane head region (TMH/Inc-region) in the Chlamydophila abortus genome by using mass spectrometry and a monoclonal antibody that reacted with the inclusion membrane. The identified protein at 32 kDa, termed Inc766, formed highly stable oligomers when solubilized in the absence of beta-mercaptoethanol. These oligomers were resistant to SDS, to heat denaturation and to 8M urea, but very sensitive to beta-mercaptoethanol, consistent with conformations resulting from protein-protein interactions stabilized through disulphide bonds. Mass spectrometry analysis of immunoprecipitated infected cell lysates indicated that a dimer at 56 kDa was the most prominent form in solution. Cross-linking with DSP provided supporting evidence for the formation of oligomers in situ. Inc766 was expressed at 20-24h post infection and its localization pattern in the extra-inclusion space was common in all C. abortus strains tested. Taken together, Inc766 displays unique biochemical and cellular features not encountered in other Incs from other Chlamydiaceae species. Future studies of the particular characteristics especially the interactive properties of Inc766 should contribute to our understanding of the relationship of the different chlamydial species with their respective hosts.

  8. Identification of immunologically relevant proteins of Chlamydophila abortus using sera from experimentally infected pregnant ewes.

    Science.gov (United States)

    Marques, P X; Souda, Puneet; O'Donovan, J; Gutierrez, J; Gutierrez, E J; Worrall, S; McElroy, M; Proctor, A; Brady, C; Sammin, D; Basset, H F; Whitelegge, Julian P; Markey, B E; Nally, J E

    2010-08-01

    Chlamydophila abortus is an intracellular pathogen and the etiological agent of enzootic abortion of ewes (EAE). C. abortus has a biphasic development cycle; extracellular infectious elementary bodies (EB) attach and penetrate host cells, where they give rise to intracellular, metabolically active reticulate bodies (RB). RB divide by binary fission and subsequently mature to EB, which, on rupture of infected cells, are released to infect new host cells. Pregnant ewes were challenged with 2 x 10(6) inclusion forming units (IFU) of C. abortus cultured in yolk sac (comprising both EB and RB). Serum samples were collected at 0, 7, 14, 21, 27, 30, 35, 40, and 43 days postinfection (dpi) and used to identify antigens of C. abortus expressed during disease. Additionally, sera from fetal lambs were collected at 30, 35, 40, and 43 dpi. All serum samples collected from experimentally infected pregnant ewes reacted specifically with several antigens of EB as determined by one-dimensional (1-D) and 2-D gel electrophoresis; reactive antigens identified by mass spectrometry included the major outer membrane protein (MOMP), polymorphic outer membrane protein (POMP), and macrophage infectivity potentiator (MIP) lipoprotein.

  9. Chlamydophila pneumoniae induces expression of Toll-like Receptor 4 and release of TNF-α and MIP-2 via an NF-κB pathway in rat type II pneumocytes

    Directory of Open Access Journals (Sweden)

    Maass Matthias

    2005-06-01

    Full Text Available Abstract Background The role of alveolar type II cells in the regulation of innate and adaptive immunity is unclear. Toll-like receptors (TLRs have been implicated in host defense. The purpose of the present study was to investigate whether Chlamydophila pneumoniae (I alters the expression of TLR2 and/orTLR4 in type II cells in a (II Rho-GTPase- and (III NF-κB-dependent pathway, subsequently (IV leading to the production of (IV pro-inflammatory TNF-α and MIP-2. Methods Isolated rat type II pneumocytes were incubated with C. pneumoniae after pre-treatment with calcium chelator BAPTA-AM, inhibitors of NF-κB (parthenolide, SN50 or with a specific inhibitor of the Rho-GTPase (mevastatin. TLR2 and TLR4 mRNA expressions were analyzed by PCR. Activation of TLR4, Rac1, RhoA protein and NF-κB was determined by Western blotting and confocal laser scan microscopy (CLSM and TNF-α and MIP-2 release by ELISA. Results Type II cells constitutively expressed TLR4 and TLR2 mRNA. A prominent induction of TLR4 but not TLR2 mRNA was detected after 2 hours of incubation with C. pneumoniae. The TLR4 protein expression reached a peak at 30 min, began to decrease within 1–2 hours and peaked again at 3 hours. Incubation of cells with heat-inactivated bacteria (56°C for 30 min significantly reduced the TLR4 expression. Treated bacteria with polymyxin B (2 μg/ml did not alter TLR4 expression. C. pneumoniae-induced NF-κB activity was blocked by TLR4 blocking antibodies. TLR4 mRNA and protein expression were inhibited in the presence of BAPTA-AM, SN50 or parthenolide. TNF-α and MIP-2 release was increased in type II cells in response to C. pneumoniae, whereas BAPTA-AM, SN50 or parthenolide decreased the C. pneumoniae-induced TNF-α and MIP-2 release. Mevastatin inhibited C. pneumoniae-mediated Rac1, RhoA and TLR4 expression. Conclusion The TLR4 protein expression in rat type II cells is likely to be mediated by a heat-sensitive C. pneumoniae protein that induces a

  10. Comparison of three different novel Chlamydophila pneumonia recombinant antigens for serodiagnosis%肺炎嗜衣原体三种不同抗原的临床诊断价值评价

    Institute of Scientific and Technical Information of China (English)

    陈虹亮; 胡旃; 周洲; 曾焱华; 代国知; 林应标; 吴移谋

    2010-01-01

    Objective To evaluate three different Chlamydophila pneumoniae recombinant antigens for use in Chlamydophila pneumoniae serodiagnosis. Methods The recombinant plasmids pGEX6p-2/ Cpn0146,Cpn0147 and Cpn0308 were constructed and expressed as GST fusion proteins. The immunogenicity and the immunocompetence of these recombinant protein were analyzed by Western-blot and indirect ELISA. A total of 183 sera samples of patients with respiratory tract infection and 32 sera samples of patients with Chlamydia trachomatis infection were detected with indirect ELISA coated microwell plates with the purified recombinant proteins comparing with SeroCP-TM IgG ELISA kits. The positive recognition rate, sensitivity and specificity of each method were analyzed. Results GST-Cpn0146, Cpn0147 and Cpn0308 were obtained after expression and purification. The titers of the specific IgG antibodies against Cpn0146, Cpn0147 and Cpn0308 were higher than 1:6 400, 1:128 00 and 1:128 00, respectively. When the indirect ELISA was developed to detect the IgG antibody against Chlamydophila pneumoniae in 183 samples, the concordance rate between the indirect ELISA test and SeroCP-TM IgG ELISA kits were 92. 3% (Cpn0146) , 94.5% (Cpn0147) and 96.7% (Cpn0308), respectively. The recombinant Cpn0146, Cpn0147 and Cpn0308 were recognized by 71 (38.8% positive recognition rate), 75 (40.9%), and 82 (44.8%) samples, respectively. The recombinant antigen-based detection assays displayed > 97% of detection specificity and>87%of sensitivity.Condusion GST-Cpn0308 shows a better sensitivity and specificity,which suggests it could be used for developing serodiagnosis kits of Chlamydophila pneumoniae infection.%目的 研究肺炎嗜衣原体3种重组抗原Cpn0146、Cpn0147及Cpn0308应用于肺炎嗜衣原体感染血清学诊断中的价值.方法 构建pGEX6p-2/Cpn0146、Cpn0147、Cpn0308重组质粒,并诱导表达重组蛋白,采用间接ELISA法和免疫印迹法(Western-blot)分析其免疫原性及其

  11. Expression of human proinflammatory cytokines from monocytic cells induced by Cpn0425 recombinant protein from Chlamydophila pneumoniae and apoptosis%Cpn0425重组蛋白诱导细胞凋亡和产生前炎症因子的研究

    Institute of Scientific and Technical Information of China (English)

    肖光文; 刘良专; 谢小平; 吴移谋

    2013-01-01

    stimulated by different concentrations of the recombinant protein of Cpn0425 from Chlamydophila pneumonioe after sonication,after removal of endotoxin by purification columns of ToxinEraser;,the expression of IL-8 and IL-1β were tested and proirferation by ELISA.Zhe inhibition of cell treated with Cpn0425 was assessed by WST-1.Cell apoptosis was detected by Annexin-V-FITC-PI.Results The recombinant protein of Cpn0425 stimulated THP-1 cell expressing the mRNA of IL-8 and IL-1 β and produce proinflamatory cytokines including IL-8 and IL-1 β in a dose and time-dependent manner,Cell proliferation of THP-1 was inhibited in a dose-dependent manner of GST-Cpn0425;The apoptosis was induced 24 hours after treatment of THP-1 cells with GST-Cpn0425,its highest apoptosis rate reached 17.76 ± 4.2%.Conclusions The recombinant protein of Cpn0425 can induce the expression of THP-1 cells and secretion of proinflammatory cytokines IL-8 and IL-1β,inhibit the proliferation and apoptosis of THP-1 cells;therefore may be an important virulence factor.

  12. Detection and Significance of Chlamydophila Pneumonia Infection in Elder Chronic Bronchitis Patients%老年慢性支气管炎患者肺炎嗜衣原体感染的检测诊断150例分析

    Institute of Scientific and Technical Information of China (English)

    潘小成

    2013-01-01

    [ObjectiveJTo explore the detection method of chlamydophila pneumonia infection and its clinical significance. [Methods]75 cases of elderly patients with chronic bronchitis as the experimental group and 75 cases of elderly healthy patients as the control group. The CPN-IgG antibody and CPN MOMP amplification of the gene were detected by ELISA and PCR assay, respectively. [Results]The positive rates of CPN MOMP and CPN-IgG were 64.0% and 89.3%, respectively, significantly higher than the control group with the values of 12.0% and 25.3% . [Conclusion] ELISA and PCR methods for detection of CPN, can improve the diagnostic accuracy of the old chronic bronchitis.%[目的]探讨老年慢性支气管炎(chronic bronchitis,CB)患者血清中肺炎嗜衣原体(chlamydophila pneumonia,CPN)感染的检测方法及其临床意义.[方法]75例老年CB患者为实验组,75例健康老年志愿者为对照组;分别采用酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)和聚合酶链反应(polymerase chain reaction,PCR)法检测研究对象的肺炎嗜衣原体IgG(chlamydophila pneumonia-IgG,CPN-IgG)和主要外膜蛋白(major outer membmne protein,MOMP):CPN MOMP也称为外膜蛋白A(存在于Cpn原体和网状体,主要包括MOMP).实验组与对照组均用了上述两种方法.[结果]实验组CPN MOMP阳性、CPN-IgG阳性的构成比分别为64.0%和89.3%,均明显高于对照组的12.0%、25.3%,差异均具有统计学意义(P<0.05).结果显示,PCR和ELISA两种检测方法均能反映发生肺炎嗜衣原体感染.[结论]ELISA和PCR法均可对CPN进行检测,两种方法同时运用能提高老年CB的临床诊断率,降低假阴性的发生,利于改善预后.

  13. 肺炎嗜衣原体CPAF诱导THP-1细胞产生前炎症细胞因子和凋亡%Chlamydial protease-like activity factor from Chlamydophila pneumoniae induced THP-1 cells produced proinflammtory cytokines and apoptosis

    Institute of Scientific and Technical Information of China (English)

    胡旃; 吴移谋; 陈虹亮; 郑江花; 周洲; 唐国芳

    2010-01-01

    Objective To express and purify Chlamydial protease-like activity factor(CPAF)from Chlamydophila pneumoniae,for investigating the effect of its recombinant protein GST-CPAF in inducing human monocytic cells to secrete proinflammatory cytokines and cell apoptosis.Methods The recom-bination expression plasmid pGEX6p-2/CPAF from Chlamydophila pneumoniae was transformed into E.coli.The recombination GST-CPAF was expressed after induction by IPTG,and purified by a agarose gel FF.Human monocytic cells were stimulated by the GST-CPAF to test the production of tumor necrosis factor a(TNF-α)and interleukin-6(IL- 6)by ELISA.Inhibition of cells proliferation with GST-CPAF was assessed by MTT.The THP-1 cell apoptosis stimulated by GST-CPAF was detected by Hoechst33258 fluorescence staining,DNA fragmentation analysis and cell apeptosis was detested bv Annexin V-FITC-propidiuum iodide (PI)staining.Results The recombination protein GST-CPAF was successfully expressed with high level in E.coli,and stimulated human monocytic cells to produce proinflammatory cytokines including TNF-α and IL-6 in a dose-and time-dependent manner.Otherwise,the GST-CPAF inhibited the growth of human monocytic cell in a dose-dependent manner.Apoptosis with nuclear chromatin fragmentation as well as cell shrinkage was observed by fluorescent staining and microscopy,DNA ladders in apoptosis cells were detected after 24 h with the GST-CPAF.Conclusion The GST-CPAF from Chlamydophila pneumoniae can induce the secretion of proinflammatory cytokines TNF-α and IL-6 by human monocytic cells,and inhibited the proliferation of THP-1 cell and apoptosis in vitro.%目的 研究肺炎嗜衣原体(Chlamydophila pneumoniae,Cpn)衣原体蛋白酶样活性因子(Chlamydial protease-like activity factor,CPAF)能否在体外诱导人单核细胞THP-1产生前炎症细胞因子和凋亡,为进一步探索Cpn感染宿主致病的分子机制提供实验依据.方法 将Cpn CPAF全基因克隆于pGEX6p-2

  14. Chlamydophila (Chlamydia) pneumoniae infection of human astrocytes and microglia in culture displays an active, rather than a persistent, phenotype.

    Science.gov (United States)

    Dreses-Werringloer, Ute; Gérard, Hervé C; Whittum-Hudson, Judith A; Hudson, Alan P

    2006-10-01

    The intracellular pathogen Chlamydia pneumoniae can cause persistent infections during which its morphologic, molecular, and pathogenic characteristics differ importantly from those of active infection. This bacterium was identified within astrocytes and microglia in the brain of late-onset Alzheimer disease patients. We investigated whether infection of these two host cell types displays an active or persistent growth phenotype. The human astrocytoma and microglioma cell lines U-87 MG and CHME-5 (respectively) and the human epithelial cell line HEp-2 were infected by the standard method with C pneumoniae strain AR-39. Cultures were harvested at 24, 48, and 72 hours postinfection and subjected to analysis of inclusion morphology. DNA and RNA were prepared from portions of each infected culture sample and analyzed for relative chromosome accumulation and presence or absence of several specific bacterial mRNAs. Astrocytes and microglial cells infected in vitro with C pneumoniae displayed inclusions that were indistinguishable from those characteristic of active infection of the standard HEp-2 host cell line. Real time polymerase chain reaction (PCR) showed that the relative accumulation of chlamydial chromosome over time during infection of these two cell lines also was virtually identical to that in actively infected HEp-2 cells. Reverse transcriptase PCR (RT-PCR) analyses showed that mRNA from ftsK, pyk, and other chlamydial genes whose expression is abrogated during persistent infection were easily identifiable in infected CHME-5 and U-87 MG cells. In cultured human astrocytes and microglia, C pneumoniae displays an active, not a persistent, growth phenotype. This indicates normal passage through the developmental cycle with its probable concomitant destruction by lysis of some portion of host cells at the termination of that cycle.

  15. 肺炎嗜衣原体菌体抗原的制备及应用%Preparation and use of Chlamydophila pneumoniae somatic antigen ELISA

    Institute of Scientific and Technical Information of China (English)

    李雪萍; 陈学勤; 钱利生

    2009-01-01

    目的 制备肺炎嗜衣原体(Chlamydophila pneumoniae,Cpn)菌体抗原,纯化后建立ELISA方法,用于血清Cpn抗体的检测. 方法 应用进口Cpn毒株感染Hep-2细胞,利用吖啶橙染色和直接免疫荧光染色判断Cpn感染细胞,制备Cpn抗原.用初步纯化的Cpn抗原建立ELISA方法检测人血清特异性抗体. 结果 吖啶橙染色和直接免疫荧光检测显示,Cpn成功感染Hep-2细胞.提取Cpn抗原,用ELISA检测84份病人血清特异抗体,阳性率为52.4%,ELISA试剂盒法的阳性率为51.2%,差异无统计学意义(P>0.05).用Cpn菌体抗原ELISA检测209份病人血清,阳性率59.3%(其中心血管疾病患者血清138份,阳性率58.0%;呼吸系统疾病患者血清71份,阳性率62.0%);检测84份健康者血清,阳性率2.4%,差异有统计学意义(P<0.05). 结论 Cpn菌体抗原ELISA检测血清抗Cpn抗体可用于判断Cpn感染;病人血清Cpn抗体阳性表明,某些心血管和呼吸系统疾病患者的致病原因可能与肺炎嗜衣原体感染有关.

  16. Recombinant 35-kDa inclusion membrane protein IncA as a candidate antigen for serodiagnosis of Chlamydophila pecorum.

    Science.gov (United States)

    Mohamad, Khalil Yousef; Rekiki, Abdessalem; Berri, Mustapha; Rodolakis, Annie

    2010-07-14

    Chlamydophila pecorum strains are commonly found in the intestine and vaginal mucus of asymptomatic ruminants and may therefore induce a positive serological response when the animals are tested for C. abortus. They have also been associated with different pathological diseases in ruminants, swine and koala. The aim of this study was to identify specific C. pecorum immunodominant antigens which could be used in ELISA tests allowing to distinguish between animals infected with C. pecorum and those infected with other chlamydial species. A gene encoding 35-kDa inclusion membrane protein incA of C. pecorum was isolated by immunoscreening of the C. pecorum DNA library using ovine anti-C. pecorum antibodies. The recombinant IncA protein did not react with a murine serum directed against C. abortus but did react with a specific monoclonal antibody of C. pecorum and toward several ovine serum samples obtained after experimental infection with different C. pecorum strains. This protein could be a good candidate for specific diagnosis of C. pecorum infection.

  17. Functional interaction between type III-secreted protein IncA of Chlamydophila psittaci and human G3BP1.

    Science.gov (United States)

    Borth, Nicole; Litsche, Katrin; Franke, Claudia; Sachse, Konrad; Saluz, Hans Peter; Hänel, Frank

    2011-01-31

    Chlamydophila (Cp.) psittaci, the causative agent of psittacosis in birds and humans, is the most important zoonotic pathogen of the family Chlamydiaceae. These obligate intracellular bacteria are distinguished by a unique biphasic developmental cycle, which includes proliferation in a membrane-bound compartment termed inclusion. All Chlamydiaceae spp. possess a coding capacity for core components of a Type III secretion apparatus, which mediates specific delivery of anti-host effector proteins either into the chlamydial inclusion membrane or into the cytoplasm of target eukaryotic cells. Here we describe the interaction between Type III-secreted protein IncA of Cp. psittaci and host protein G3BP1 in a yeast two-hybrid system. In GST-pull down and co-immunoprecipitation experiments both in vitro and in vivo interaction between full-length IncA and G3BP1 were shown. Using fluorescence microscopy, the localization of G3BP1 near the inclusion membrane of Cp. psittaci-infected Hep-2 cells was demonstrated. Notably, infection of Hep-2 cells with Cp. psittaci and overexpression of IncA in HEK293 cells led to a decrease in c-Myc protein concentration. This effect could be ascribed to the interaction between IncA and G3BP1 since overexpression of an IncA mutant construct disabled to interact with G3BP1 failed to reduce c-Myc concentration. We hypothesize that lowering the host cell c-Myc protein concentration may be part of a strategy employed by Cp. psittaci to avoid apoptosis and scale down host cell proliferation.

  18. BLAST screening of chlamydial genomes to identify signature proteins that are unique for the Chlamydiales, Chlamydiaceae, Chlamydophila and Chlamydia groups of species

    Directory of Open Access Journals (Sweden)

    Gupta Radhey S

    2006-01-01

    Full Text Available Abstract Background Chlamydiae species are of much importance from a clinical viewpoint. Their diversity both in terms of their numbers as well as clinical involvement are presently believed to be significantly underestimated. The obligate intracellular nature of chlamydiae has also limited their genetic and biochemical studies. Thus, it is of importance to develop additional means for their identification and characterization. Results We have carried out analyses of available chlamydiae genomes to identify sets of unique proteins that are either specific for all Chlamydiales genomes, or different Chlamydiaceae family members, or members of the Chlamydia and Chlamydophila genera, or those unique to Protochlamydia amoebophila, but which are not found in any other bacteria. In total, 59 Chlamydiales-specific proteins, 79 Chlamydiaceae-specific proteins, 20 proteins each that are specific for both Chlamydia and Chlamydophila and 445 ORFs that are Protochlamydia-specific were identified. Additionally, 33 cases of possible gene loss or lateral gene transfer were also detected. Conclusion The identified chlamydiae-lineage specific proteins, many of which are highly conserved, provide novel biomarkers that should prove of much value in the diagnosis of these bacteria and in exploration of their prevalence and diversity. These conserved protein sequences (CPSs also provide novel therapeutic targets for drugs that are specific for these bacteria. Lastly, functional studies on these chlamydiae or chlamydiae subgroup-specific proteins should lead to important insights into lineage-specific adaptations with regards to development, infectivity and pathogenicity.

  19. Genetic variability of Chlamydophila abortus strains assessed by PCR-RFLP analysis of polymorphic membrane protein-encoding genes.

    Science.gov (United States)

    Sait, Michelle; Clark, Ewan M; Wheelhouse, Nicholas; Spalding, Lucy; Livingstone, Morag; Sachse, Konrad; Markey, Bryan K; Magnino, Simone; Siarkou, Victoria I; Vretou, Evangelia; Caro, María R; Yaga, Raja; Lainson, F Alex; Smith, David G E; Wright, Frank; Longbottom, David

    2011-08-05

    This study used PCR-RFLP to investigate the genetic variability of pmp-encoding genes from fifty-two Chlamydophila abortus (C. abortus) strains originating from abortion cases from various geographical regions and host species. Six primer pairs were used to PCR-amplify DNA fragments encoding eighteen pmps. PCR products were digested using four restriction endonucleases and Bayesian methodologies were used to compare RFLP profiles and assign strains to a RFLP genotype. Strains could be assigned to 2 genotypes in the region encoding pmp18D, 3 genotypes in the regions encoding pmp1A-pmp2B, pmp3E-pmp6H and pmp11G-pmp15G, 4 genotypes in the region encoding pmp7G-pmp10G and 5 genotypes in the region encoding pmp16G-pmp17G. In all regions, the majority of strains (88.4-96.1%) had the same genotype as the reference strain S26/3. No correlation could be made between genotype, host species or geographical origin except for the two variant Greek strains, LLG and POS, which formed a discrete genotype in all pmp-encoding regions except pmp18D. Relative rates of evolution calculated for each pmp-encoding gene locus suggest that differing selective pressures and functional constraints may exist on C. abortus polymorphic membrane proteins. These findings suggest that although intraspecies heterogeneity of pmp-encoding genes in C. abortus is low, the sequence heterogeneity should be an important consideration when using pmps as the basis for novel diagnostics or vaccine development.

  20. Protection of mice against Chlamydophila abortus infection with a bacteriophage-mediated DNA vaccine expressing the major outer membrane protein.

    Science.gov (United States)

    Ling, Yong; Liu, Wei; Clark, Jason R; March, John B; Yang, Junjing; He, Cheng

    2011-12-15

    A bacteriophage-delivered DNA vaccine against Chlamydophila abortus was constructed by cloning a eukaryotic cassette containing the ompA gene (which expresses the Major Outer Membrane Protein) into a bacteriophage lambda vector. Four groups, each of 20 BALB/c mice were inoculated separately with the phage vaccine, a conventional DNA vaccine based on the same ompA expression cassette, a live attenuated vaccine (strain 1B) or the empty phage vector. The phage and DNA vaccines and empty phage vector were administered intramuscularly on days 0, 14 and 28; the attenuated vaccine was given once on day 0. Half the animals in each group were challenged on day 42 by intraperitoneal injection of live C. abortus and sacrificed on day 49. Phage-vaccinated mice developed moderate antibody levels against C. abortus and yielded higher levels of IFN-γ and IL-2 compared with the attenuated live vaccine group. Clearance of chlamydiae from spleens was significantly better in the attenuated vaccine group compared with the phage vaccine group, while both groups were significantly superior to the DNA vaccine and control groups (p<0.01). Although levels of protection in the mouse model were lower in phage-vaccinated animals, than in 1B vaccinated animals, phage vaccines offer several other advantages, such as easier handling and safety, potentially cheaper production and no chance of reversion to virulence. Although these are preliminary results in a model system, it is possible that with further optimisation immunization with phage vaccines may provide a novel way to improve protection against C. abortus infection and trials in large animals are currently being initiated.

  1. Chlamydophila pneumonia-Derived Inclusion Membrane Protein Cpn0147 Induces Monocytes Secretion of TNF-αand IL-8%肺炎嗜衣原体包涵体膜蛋白Cpn0147诱导单核细胞分泌TNF-α和IL-8

    Institute of Scientific and Technical Information of China (English)

    吴华; 卿文衡; 刘军; 张黎黎

    2015-01-01

    Objective To investigate the molecular mechanism of the inclusion membrane protein Cpn0147 on the se-cretion of cytokines in monocytes. Methods Cultured THP-1 cells were stimulated with different concentration of endotox-in-free recombinant inclusion protein (1. 0,10. 0,30. 0 and 50. 0μg/mL) for 0~48 h. Secretion of TNF-αand IL-8 were de-tected by ELISA,expression of the mRNA was measured by real-time PCR. In addition,cells were preincubated with Toll-like receptor 2(TLR2) or TLR4 neutralizing antibody,or transfected with siRNA for TLR2 and TLR4,secretion of TNF-αand IL-8 were detected by ELISA. Results Cpn0147 could induce THP-1 cells the secretion of TNF-αand IL-8,and the expres-sion of mRNA. The production of TNF-αand IL-8 varies with time intervals,the ELISA results indicated secretion of cytokines appeared after 2~6 h of stimulation,peaked at 12 h and then decreased. Pre-incubation of TLR2 or TLR4 neutralizing anti-body significantly abrogate Cpn0147-induced cytokines production,similar results was also obtained by RNA interference of TLR2 and TLR4. Conclusion Cpn0147 can induce TNF-αand IL-8 secretion via TLR2 and TLR4.%目的:探讨肺炎嗜衣原体( Cpn)包涵体膜蛋白Cpn0147诱导人单核细胞分泌炎症因子的分子机制。方法用去除内毒素活性的不同浓度Cpn0147重组蛋白(1.0、10.0、30.0和50.0μg/mL)刺激THP-1细胞0~48 h,ELISA检测肿瘤坏死因子-α(TNF-α)和白介素-8(IL-8)的分泌水平;实时定量PCR检测TNF-α和IL-8 mRNA的表达;用Toll样受体2( TLR2)和TLR4中和抗体处理THP-1细胞,或采用 TLR2和 TLR4 siRNA沉默其表达, ELISA检测THP-1处理前后TNF-α和IL-8分泌的变化。结果 Cpn0147可诱导THP-1细胞表达TNF-α和IL-8的mRNA和蛋白;同时,不同时间点Cpn0147对TNF-α和IL-8的诱导水平有所不同,Cpn0147作用2~6 h后即可诱导TNF-α和IL-8分泌,12 h时达到峰值,随后逐渐下降。采用TLR2和TLR4中和抗体封闭THP-1细胞后,TNF-α和IL-8分

  2. Application of nucleic acid amplification technique for diagnosis of Chlamydophila pneumonia infection%核酸扩增技术在肺炎嗜衣原体感染诊断中的应用

    Institute of Scientific and Technical Information of China (English)

    张军华; 陈丽丽; 吴移谋

    2010-01-01

    Chlamydia pneumoniae (Cpn) is a kind of microorganism parasitizing in eukaryotic cells, causing human respiratory tract infection, and having persistent infection in the body. Rapid diagnosis in the early stage of Cpn infection helps to prevent the spread of disease and the formation of complications. On the Cpn diagnostic methods, domestic and foreign scholars have carried out a series of studys and made great progress. This paper reviews the application of nucleic acid amplification technique for the diagnosis of Cpn infection.%肺炎嗜衣原体(Chlamydophila pneumonia,Cpn)是一类引起人类呼吸道感染的、专营真核细胞内寄生生活的微生物,在人体中存在持续感染.Cpn感染早期的快速诊断有利于阻止疾病的传播和防止并发症的形成.有关Cpn的诊断方法,国内外学者进行了一系列研究并取得了很大的进展.本文就核酸扩增技术在诊断Cpn感染中的应用作一综述.

  3. A Cluster of Legionella-Associated Pneumonia Cases in a Population of Military Recruits

    Science.gov (United States)

    2007-06-01

    pneumophila, Mycoplasma pneumoniae, Chlamydophila pneumoniae, and Bordetella pertussis (8). This test was applied retrospectively to 240 samples... Chlamydophila ) pneumoniae, Legionella pneumophila, Legionella micdadei, and Bordetella pertussis, and its real-time counterpart. J. Clin. Microbiol. 43:565–571...8. McDonough, E. A., C. P. Barrozo, K. L. Russell, and D. Metzgar. 2005. A multiplex PCR for detection of Mycoplasma pneumoniae, Chlamydophila

  4. Zoonotic potential of Chlamydophila.

    Science.gov (United States)

    Rodolakis, Annie; Yousef Mohamad, Khalil

    2010-01-27

    The purpose of this article is to present the diseases induced in humans and animals by the different species of Chlamydophila, after providing an overview on the history of these infectious agents and their taxonomy. The route of transmission and the available methods for prevention and control in the different animal species are reviewed.

  5. 呼吸道感染患者外周血中肺炎嗜衣原体的检测与分析%Detection and analysis for Chlamydophila pneumoniae in peripheral blood in patients with respiratory tract infection

    Institute of Scientific and Technical Information of China (English)

    吴华; 吴移谋; 黄志坚; 詹济舟; 刘良专

    2011-01-01

    In order to investigate the infection rate of Chlamydophila pneumoniae, blood samples were collected from 173 respiratory tract infection patients and 52 healthy volunteers. DNA was extracted from PBMC and sera were isolated from the blood samples. The MOMP gene of C. pneumoniae were detected by PCR from the DNA samples and the C. pneumoniae specific IgG were measured by ELISA. PCR products were detected by 1.5% agarose gel electrophoresis and sequenced by Sangon Company. Combined with the clinic data, the difference of C. pneumoniae infected rate in different age group were analysed. The results of PBMC extraction were as follow: there're 173 people in the respiratory tract infection group, and the PCR (MOMP) positive rate was 32.40%(56/173); there're 52 people in the healthy group, and the PCR(MOMP) positive rate was 9.62% (5/52). The MOMP PCR product with Cpn AR-39 of the MOMP gene was 99% consistent. According to the positive rate of each group, the C. pneumoniae infection rate in the aged group was the highest. It's implied that Cpn is one of the most important respiratory tract pathogen and proved to be the main pathogen for C. pneumoniae. The older people are in highrisk group for C. pneumoniae infection.%目的 检测并分析呼吸道疾病患者的肺炎嗜衣原体(Cpn)感染率.方法 收集173例呼吸道感染患者和52例健康体检者外周血标本,分离外周血单核细胞(PBMC)血清.提取纯化PBMC的DNA,以该DNA为模板,用特异性引物体外扩增Cpn MOMP基因.扩增产物经电泳及测序鉴定,确认为Cpn特异性的目的 基因.用ELISA法检测待测血清中的Cpn-IgG抗体.通过PCR与ELISA的检测结果,结合临床资料进行流行病学综合分析Cpn感染率.结果 以PBMC基因组DNA为模板,呼吸道感染组患者标本中Cpn MOMP阳性率为32.40%(56/173),健康对照组中阳性率为9.62 oA(5/52);经测序比对,所有MOMP阳性扩增产物与Cpn AR-39的MOMP基因同源性在99%以上.ELISA检测血

  6. Pneumonia

    Science.gov (United States)

    ... of pneumonia. Be sure to get the following vaccines: Flu vaccine can help prevent pneumonia caused by the flu virus. Pneumococcal vaccine lowers your chances of getting pneumonia from Streptococcus ...

  7. Pneumonia

    Science.gov (United States)

    ... overall health and whether it's caused by a virus or bacteria. With pneumonia caused by bacteria, a kid might ... tell if the infection is caused by a virus or bacteria. previous continue No More Pneumonia If the pneumonia ...

  8. Genome sequence of the Chlamydophila abortus variant strain LLG.

    Science.gov (United States)

    Sait, Michelle; Clark, Ewan M; Wheelhouse, Nick; Livingstone, Morag; Spalding, Lucy; Siarkou, Victoria I; Vretou, Evangelia; Smith, David G E; Lainson, F Alex; Longbottom, David

    2011-08-01

    Chlamydophila abortus is a common cause of ruminant abortion. Here we report the genome sequence of strain LLG, which differs genotypically and phenotypically from the wild-type strain S26/3. Genome sequencing revealed differences between LLG and S26/3 to occur in pseudogene content, in transmembrane head/inc family proteins, and in biotin biosynthesis genes.

  9. 肺炎嗜衣原体MOMP基因的克隆与原核表达%Cloning and Prokaryotic Expression of the MOMP Gene of Chlamydophila Pneumoniae

    Institute of Scientific and Technical Information of China (English)

    张存亮; 聂福平; 王昱; 杨俊; 李应国; 蔡家利

    2014-01-01

    克隆了肺炎嗜衣原体外膜主蛋白(major out membrane protein,MOMP)基因,并进行了原核表达.根据GenBank公布的肺炎嗜衣原体MOMP基因序列,设计克隆引物后用普通PCR方法扩增出肺炎嗜衣原体MOMP基因的完整片段,将其克隆到pMD-19T后进行测序,经序列比对正确后将此片段亚克隆到表达载体pET-32a(+)中,转化至大肠杆菌Rosetta(DE3),经IPTG诱导表达,再采用SDS-PAGE和Western-Blot检测重组蛋白的表达情况.结果表明:本研究克隆了肺炎嗜衣原体MOMP基因并在原核系统中成功表达了MOMP重组蛋白,且Western-Blot显示该重组蛋白具有免疫学活性.

  10. Study on the diagnosis application using recombinant protein of immunodominant region gene of chlamydial protease-like activity factor from Chlamydophila psittaci%鹦鹉热嗜衣原体蛋白酶样活性因子免疫优势区基因重组蛋白的诊断应用研究

    Institute of Scientific and Technical Information of China (English)

    王绍胜; 吴移谋; 陈丽丽; 曾焱华; 刘良专

    2011-01-01

    目的 检测鹦鹉热嗜衣原体(Chlamydophila psittaci,Cps)蛋白酶样活性因子(chlamydial protease-like activity factor,CPAF)免疫优势区基因在E.coli BL21中高效表达的产物在Cps感染诊断中的应用,为建立Cps感染的快速诊断奠定实验基础.方法 利用生物信息学软件筛选出Cps CPAF免疫优势区基因序列(CPAFm,A196~A450),设计特异性引物,PCR扩增目的基因,将其克隆入pGEX6p-2载体后转化E.coli BL21,利用IPTG诱导表达重组蛋白,Western blot鉴定重组蛋白.以纯化的重组蛋白作为包被抗原,建立血清学诊断的间接ELISA法;同时用商品ELISA试剂盒与建立的间接ELISA法检测180份可疑Cps感染的病鸭血清标本,将检测结果进一步用Western blot方法验证.结果 构建原核重组质粒pGEX6p-2/CpsCPAFm,表达相对分子质量约为54×103的目的蛋白产物.以纯化的重组蛋白为包被抗原建立间接ELISA法检测Cps参考血清,其阴、阳性符合率均为100%;与肺炎嗜农原体(C.pneumoniae,Cpn)、沙眼衣原体(C.trachomatis,Ct)无交叉反应.对180份可疑病鸭血清标本进行检测,与商品Birds Chlamydia Psittaci IgG ELISA试剂盒比较,以Western blot方法作对照,自建的ELISA法符合率均为100%,Birds Chlamydia Psittaci IgG ELISA Kit符合率为77.5%~95.0%.结论 以Cps CPAF免疫优势区(A196~A450)作为诊断抗原,建立血清学诊断的间接ELISA法具有较好的临床诊断价值.%Objective To clone and express the immunodominant domain of the chlamydial proteaselike activity factor(CPAF) from Chlamydophila psittaci(Cps) and evaluated the diagnosing value of the recombinant protein in Cps infection.Methods The immunodominant region epitope of CPAF (CPAFm,A196-A450)from Cps was chosen according to bioinformatics analysis and references.The specific primer was designed and the gene was amplified by PCR and then ligated into a pGEX6p-2 vector.Recombinant protein was induced to express by IPTG and

  11. Kadar Protein Klebsiella pneumoniae Hasil Pemanasan 65 Derajat Celcius

    Directory of Open Access Journals (Sweden)

    Irawan Sugoro

    2016-03-01

    Full Text Available Klebsiella pneumoniae is one of a coliform bacteria that causing mastitis. This disease were founded in dairy cows and can be prevented by vaccination. The research has been conducted to determine the inactive times, the protein concentration and profile of K. pneumoniae which inactivated by heating of 65oC as material of mastitis vaccine. The cells culture inactivated by the different times, i.e. 0, 10, 20, 30, 40, 50 and 60 minutes. The inactive times was determined by the drop test method, whereas the protein concentration of cells were determined by Lowry method. The results showed that the inactive times occured after 30 minute, and has a significant different on the protein concentration of bacteria cells that inactivated by the different times.

  12. Identification of antigenic proteins of the nosocomial pathogen Klebsiella pneumoniae.

    Directory of Open Access Journals (Sweden)

    Sebastian Hoppe

    Full Text Available The continuous expansion of nosocomial infections around the globe has become a precarious situation. Key challenges include mounting dissemination of multiple resistances to antibiotics, the easy transmission and the growing mortality rates of hospital-acquired bacterial diseases. Thus, new ways to rapidly detect these infections are vital. Consequently, researchers around the globe pursue innovative approaches for point-of-care devices. In many cases the specific interaction of an antigen and a corresponding antibody is pivotal. However, the knowledge about suitable antigens is lacking. The aim of this study was to identify novel antigens as specific diagnostic markers. Additionally, these proteins might be aptly used for the generation of vaccines to improve current treatment options. Hence, a cDNA-based expression library was constructed and screened via microarrays to detect novel antigens of Klebsiella pneumoniae, a prominent agent of nosocomial infections well-known for its extensive antibiotics resistance, especially by extended-spectrum beta-lactamases (ESBL. After screening 1536 clones, 14 previously unknown immunogenic proteins were identified. Subsequently, each protein was expressed in full-length and its immunodominant character examined by ELISA and microarray analyses. Consequently, six proteins were selected for epitope mapping and three thereof possessed linear epitopes. After specificity analysis, homology survey and 3d structural modelling, one epitope sequence GAVVALSTTFA of KPN_00363, an ion channel protein, was identified harboring specificity for K. pneumoniae. The remaining epitopes showed ambiguous results regarding the specificity for K. pneumoniae. The approach adopted herein has been successfully utilized to discover novel antigens of Campylobacter jejuni and Salmonella enterica antigens before. Now, we have transferred this knowledge to the key nosocomial agent, K. pneumoniae. By identifying several novel antigens

  13. Identification of Antigenic Proteins of the Nosocomial Pathogen Klebsiella pneumoniae

    Science.gov (United States)

    Hoppe, Sebastian; Bier, Frank F.; von Nickisch-Rosenegk, Markus

    2014-01-01

    The continuous expansion of nosocomial infections around the globe has become a precarious situation. Key challenges include mounting dissemination of multiple resistances to antibiotics, the easy transmission and the growing mortality rates of hospital-acquired bacterial diseases. Thus, new ways to rapidly detect these infections are vital. Consequently, researchers around the globe pursue innovative approaches for point-of-care devices. In many cases the specific interaction of an antigen and a corresponding antibody is pivotal. However, the knowledge about suitable antigens is lacking. The aim of this study was to identify novel antigens as specific diagnostic markers. Additionally, these proteins might be aptly used for the generation of vaccines to improve current treatment options. Hence, a cDNA-based expression library was constructed and screened via microarrays to detect novel antigens of Klebsiella pneumoniae, a prominent agent of nosocomial infections well-known for its extensive antibiotics resistance, especially by extended-spectrum beta-lactamases (ESBL). After screening 1536 clones, 14 previously unknown immunogenic proteins were identified. Subsequently, each protein was expressed in full-length and its immunodominant character examined by ELISA and microarray analyses. Consequently, six proteins were selected for epitope mapping and three thereof possessed linear epitopes. After specificity analysis, homology survey and 3d structural modelling, one epitope sequence GAVVALSTTFA of KPN_00363, an ion channel protein, was identified harboring specificity for K. pneumoniae. The remaining epitopes showed ambiguous results regarding the specificity for K. pneumoniae. The approach adopted herein has been successfully utilized to discover novel antigens of Campylobacter jejuni and Salmonella enterica antigens before. Now, we have transferred this knowledge to the key nosocomial agent, K. pneumoniae. By identifying several novel antigens and their linear

  14. Pneumonia

    Science.gov (United States)

    ... better than treating it. Vaccines are available to prevent pneumococcal pneumonia and the flu. Other preventive measures include washing your hands frequently and not smoking. NIH: National Heart, Lung, and Blood Institute

  15. Anticuerpos contra Chlamydophilaen pacientes con infarto agudo del miocardio y riesgo coronario, y su relación con la muerte Antibodies against Chlamydophila in patients with acute myocardial infarction and coronary risk and their association with mortality

    Directory of Open Access Journals (Sweden)

    Guadalupe García-Elorriaga

    2005-06-01

    Full Text Available OBJETIVO: Determinar si los anticuerpos contra Chlamydophila pneumoniae en pacientes con infarto agudo del miocardio y factores de riesgo coronario se asocian con la muerte. MATERIAL Y MÉTODOS: Se hizo un estudio observacional, prospectivo, transversal y comparativo. Se incluyeron en el estudio 100 sujetos que, entre 1999 y 2000, estuvieron hospitalizados en la Unidad Coronaria del Hospital de Especialidades del Centro Médico La Raza, del Instituto Mexicano del Seguro Social. Se trataba de una muestra constituida por pacientes de ambos sexos, mayores de 18 años, con infarto agudo del miocardio y riesgo coronario. Mediante microinmunofluorescencia indirecta se identificaron anticuerpos contra Chlamydophila pneumoniae, Chlamydophila psitacii y Chlamydia trachomatis. De entre los 100 sujetos, se eligieron al azar 33, a quienes se les determinaron anticuerpos contra Chlamydophila, no sólo durante su estancia en el hospital, sino también al salir de éste y a los tres meses de haber sufrido el infarto agudo del miocardio. Se calcularon las medias y las desviaciones geométricas estándares para los títulos de anticuerpos contra Chlamydophila, y se determinó la razón de momios y el intervalo de confianza al 95% entre los factores de riesgo coronario y la muerte. RESULTADOS: Setenta por ciento de los pacientes de la muestra inicial presentaron anticuerpos contra Chlamydophila pneumoniae; no se identificaron anticuerpos contra Chlamydophila psitacii y Chlamydia trachomatis. No se observó una fuerza de asociación estadísticamente significativa con la muerte en pacientes con infarto agudo del miocardio y factores de riesgo coronario. De los 33 individuos de la submuestra, 25 presentaron anticuerpos contra Chlamydophila pneumoniae, y en 83% de estos últimos casos, se registró un descenso de dichos anticuerpos a los tres meses de haberse presentado el infarto agudo del miocardio. CONCLUSIONES: A pesar de que en pacientes con infarto agudo del

  16. Peritoneal culture alters Streptococcus pneumoniae protein profiles and virulence properties

    Science.gov (United States)

    Orihuela, C. J.; Janssen, R.; Robb, C. W.; Watson, D. A.; Niesel, D. W.

    2000-01-01

    We have examined the properties of Streptococcus pneumoniae cultured in the murine peritoneal cavity and compared its virulence-associated characteristics to those of cultures grown in vitro. Analysis of mRNA levels for specific virulence factors demonstrated a 2.8-fold increase in ply expression and a 2.2-fold increase in capA3 expression during murine peritoneal culture (MPC). Two-dimensional gels and immunoblots using convalescent-phase patient sera and murine sera revealed distinct differences in protein production in vivo (MPC). MPC-grown pneumococci adhered to A549 epithelial cell lines at levels 10-fold greater than those cultured in vitro.

  17. Improved sensitivity of PCR for Chlamydophila using pmp genes.

    Science.gov (United States)

    Laroucau, K; Souriau, A; Rodolakis, A

    2001-09-20

    Primers targeting the conserved pmp gene family of Chlamydophila abortus were evaluated for their ability to improve the polymerase chain reaction (PCR) sensitivity. In purified DNA, specific pmp primers (named CpsiA and CpsiB) allowed at least a 10-fold increase of the PCR sensitivity compared to the specific ompA primers for C. abortus, but also for C. psittaci and C. caviae strains. No amplification was observed on C. felis, C. pecorum, C. pneumoniae and Chlamydia trachomatis strains. Tested on contaminated specimens such as genital swabs, the PCR sensitivity observed with CpsiA/CpsiB was also better than with the ompA primers. This study demonstrated that these specific pmp primers could serve as valuable, sensitive and common tools for a specific Chlamydophila diagnosis in ruminant, avian and human diseases. Digestion by AluI of the CpsiA/CpsiB fragments allowed a specific discrimination of the strains in function of their hosts and/or their serotypes.

  18. Therapeutic Chlamydophila abortus and C. pecorum vaccination transiently reduces bovine mastitis associated with Chlamydophila infection.

    Science.gov (United States)

    Biesenkamp-Uhe, Carolin; Li, Yihang; Hehnen, Hans-Robert; Sachse, Konrad; Kaltenboeck, Bernhard

    2007-02-01

    Infections with Chlamydophila abortus and C. pecorum are highly prevalent in cattle and have been associated with bovine mastitis. A prospective cohort study was conducted with a herd of 140 Holstein dairy cows to investigate the influence of Chlamydophila infection on subclinical inflammation of the bovine mammary gland as characterized by somatic cell numbers in milk. PCR detection of C. abortus and low serum antibody levels against Chlamydophila spp. were significantly associated with subclinical mastitis. To examine the effect of the infection by response modification, immune perturbation was done by two subcutaneous administrations of an experimental vaccine preparation of inactivated C. abortus and C. pecorum elementary bodies. Vaccination against Chlamydophila highly significantly decreased milk somatic cell numbers, thus reducing bovine mastitis, and increased antibody levels against Chlamydophila but did not eliminate shedding of C. abortus in milk as detected by PCR. The protective effect peaked at 11 weeks after vaccination and lasted for a total of 14 weeks. Vaccination with the Chlamydophila vaccine, a mock vaccine, or a combination vaccine against bovine viral diseases highly significantly increased C. abortus shedding in milk for 1 week, presumably mediated by the vaccine adjuvant. In summary, this study shows an etiological involvement of the widespread Chlamydophila infections in bovine mastitis, a herd disease of critical importance for the dairy industry. Furthermore, this investigation shows the potential for temporary improvement of chlamydial disease by therapeutic vaccination. Chlamydophila vaccination of cattle might serve as a testing ground for vaccines against human chlamydial infections.

  19. High Prevalence of Natural Chlamydophila Species Infection in Calves

    OpenAIRE

    Jee, JunBae; DeGraves, Fred J.; Kim, TeaYoun; Kaltenboeck, Bernhard

    2004-01-01

    We investigated the acquisition and prevalence of Chlamydophila sp. infection in calves. Specimens were collected at weekly intervals from birth to week 12 postpartum from 40 female Holstein calf-dam pairs in a dairy herd. Real-time PCR detected, quantified, and differentiated Chlamydophila 23S rRNA gene DNA from vaginal cytobrush swabs and milk samples. Chemiluminescence enzyme-linked immunosorbent assay with lysed Chlamydophila abortus or Chlamydophila pecorum elementary body antigens quant...

  20. OmpA and antigenic diversity of bovine Chlamydophila pecorum strains.

    Science.gov (United States)

    Kaltenboeck, B; Heinen, E; Schneider, R; Wittenbrink, M M; Schmeer, N

    2009-03-16

    Infections with the intracellular bacterium Chlamydophila (C.) pecorum are highly prevalent worldwide in cattle. These infections cause significant diseases such as polyarthritis, pneumonia, enteritis, genital infections and fertility disorders, and occasionally sporadic bovine encephalomyelitis. Subclinical respiratory infections of calves with C. pecorum have been associated with airway obstruction, pulmonary inflammation, and reduced weight gains. This investigation examined four chlamydial strains with biological properties of C. pecorum isolated from feces of clinically normal cattle, from calves with pneumonia, and from bulls with posthitis. The objective was to characterize the evolutionary relationships of these bovine chlamydial isolates to other chlamydiae by genetic analysis of the ompA gene, and by the immunological cross-reactivities in Western immunoblot analysis. PCR typing of the ompA gene identified these isolates as C. pecorum. The OmpA-deduced amino acid dissimilarities between these four strains spanned 10-20%. In phylogenetic analysis, the four isolates clustered with C. pecorum ruminant, porcine, and koala strains of different geographic origins rather than with each other. All four isolates showed different patterns of Western immunoblot reactivity with antiserum against bovine C. pecorum strain LW63, and, interestingly, no cross-reactivity of the OmpA proteins with the anti-LW613 OmpA antibodies. These data underscore the polyphyletic population structure of C. pecorum and suggest that the spectrum of C. pecorum OmpA proteins in a host species can occupy the entire evolutionary bandwidth within C. pecorum. The variant immunoblot reactivities support the notion of considerable genomic plasticity of C. pecorum.

  1. A protein-based pneumococcal vaccine protects rhesus macaques from pneumonia after experimental infection with Streptococcus pneumoniae.

    Science.gov (United States)

    Denoël, Philippe; Philipp, Mario T; Doyle, Lara; Martin, Dale; Carletti, Georges; Poolman, Jan T

    2011-07-26

    Infections caused by Streptococcus pneumoniae are a major cause of mortality throughout the world. Protein-based pneumococcal vaccines are envisaged to replace or complement the current polysaccharide-based vaccines. In this context, detoxified pneumolysin (dPly) and pneumococcal histidine triad protein D (PhtD) are two potential candidates for incorporation into pneumococcal vaccines. In this study, the protective efficacy of a PhtD-dPly vaccine was evaluated in a rhesus macaque (Macaca mulatta) model of pneumonia. The animals were immunized twice with 10 μg of PhtD and 10 μg of dPly formulated in the Adjuvant System AS02 or with AS02 alone, before they were challenged with a 19F pneumococcal strain. The survival was significantly higher in the protein-vaccinated group and seemed to be linked to the capacity to greatly reduce bacterial load within the first week post-challenge. Vaccination elicited high concentrations of anti-PhtD and anti-Ply antibodies and a link was found between survival and antibody levels. In conclusion, AS02-adjuvanted PhtD-dPly vaccine protects against S. pneumoniae-induced pneumonia. It is probable that the protection is at least partially mediated by PhtD- and Ply-specific antibodies.

  2. Heat-shock protein ClpL/HSP100 increases penicillin tolerance in Streptococcus pneumoniae.

    Science.gov (United States)

    Tran, Thao Dang-Hien; Kwon, Hyog-Young; Kim, Eun-Hye; Kim, Ki-Woo; Briles, David E; Pyo, Suhkneung; Rhee, Dong-Kwon

    2011-01-01

    Penicillin resistance and tolerance has been an increasing threat to the treatment of pneumococcal pneumoniae. However, no penicillin tolerance-related genes have been claimed. Here we show that a major heat shock protein ClpL/HSP100 could modulate the expression of a cell wall synthesis enzyme PBP2x, and subsequently increase cell wall thickness and penicillin tolerance in Streptococus pneumoniae.

  3. Identification of pneumococcal surface protein A as a lactoferrin-binding protein of Streptococcus pneumoniae.

    Science.gov (United States)

    Hammerschmidt, S; Bethe, G; Remane, P H; Chhatwal, G S

    1999-04-01

    Lactoferrin (Lf), an iron-sequestering glycoprotein, predominates in mucosal secretions, where the level of free extracellular iron (10(-18) M) is not sufficient for bacterial growth. This represents a mechanism of resistance to bacterial infections by prevention of colonization of the host by pathogens. In this study we were able to show that Streptococcus pneumoniae specifically recognizes and binds the iron carrier protein human Lf (hLf). Pretreatment of pneumococci with proteases reduced hLf binding significantly, indicating that the hLf receptor is proteinaceous. Binding assays performed with 63 clinical isolates belonging to different serotypes showed that 88% of the tested isolates interacted with hLf. Scatchard analysis showed the existence of two hLf-binding proteins with dissociation constants of 5.7 x 10(-8) and 2.74 x 10(-7) M. The receptors were purified by affinity chromatography, and internal sequence analysis revealed that one of the S. pneumoniae proteins was homologous to pneumococcal surface protein A (PspA). The function of PspA as an hLf-binding protein was confirmed by the ability of purified PspA to bind hLf and to competitively inhibit hLf binding to pneumococci. S. pneumoniae may use the hLf-PspA interaction to overcome the iron limitation at mucosal surfaces, and this might represent a potential virulence mechanism.

  4. Interaction between the P1 protein of Mycoplasma pneumoniae and receptors on HEp-2 cells

    DEFF Research Database (Denmark)

    Drasbek, Mette; Christiansen, Gunna; Drasbek, Kim Ryun;

    2007-01-01

    The human pathogen Mycoplasma pneumoniae can cause atypical pneumonia through adherence to epithelial cells in the respiratory tract. The major immunogenic protein, P1, participates in the attachment of the bacteria to the host cells. To investigate the adhesion properties of P1, a recombinant...... protein (rP1-II) covering amino acids 1107-1518 of the P1 protein was produced. This protein inhibited the adhesion of M. pneumoniae to human HEp-2 cells, as visualized in a competitive-binding assay using immunofluorescence microscopy. Previous studies have shown that mAbs that recognize two epitopes...... intensity. The number of M. pneumoniae microcolonies adhering to the host cells was significantly reduced by these five peptides. Further investigations showed that inhibiting peptide 7 (amino acids 1347-1396) of the major adhesin protein P1 bound directly to host receptors, suggesting that the observed M...

  5. Chlamydophila felis: plasmid detection in Italian isolates.

    Science.gov (United States)

    Di Francesco, Antonietta; Donati, Manuela; Salvatore, Daniela; Cevenini, Roberto; Di Paolo, Maria; Baldelli, Raffaella

    2010-04-01

    Plasmids have been detected in the majority of strains in the genus Chlamydia and in many Chlamydophila species. Previous studies showed that FP Pring and FP Cello Chlamydophila felis strains have an extrachromosomial plasmid, whereas the FP Baker strain does not. Azuma et al. recently sequenced the entire genomic DNA sequence of the Japanese Cp. felis strain Fe/C-56 and described a 7,552 base pair circular plasmid. In the present study a highly conserved plasmid gene was detected in 11 Italian Cp. felis isolates, showing 100% nucleotide identity with the plasmid gene of Fe/C-56 Cp. felis strain.

  6. Differential protein expression in phenotypic variants of Streptococcus pneumoniae

    NARCIS (Netherlands)

    K. Overweg (Karin); C.D. Pericone; G.G. Verhoef; J.N. Weiser; H.D. Meiring; A.P. de Jong; R. de Groot (Ronald); P.W.M. Hermans (Peter)

    2000-01-01

    textabstractStreptococcus pneumoniae undergoes spontaneous phase variation resulting in opaque and transparent colony forms. Differences in colony opacity correlate with differences in virulence: the transparent variants are more capable of colonizing the nasopharynx, w

  7. Comparative evaluation of eight serological assays for diagnosing Chlamydophila abortus infection in sheep.

    Science.gov (United States)

    Wilson, Kim; Livingstone, Morag; Longbottom, David

    2009-03-16

    Chlamydophila abortus is one of the principal causes of late-term abortion (enzootic abortion of ewes or EAE) in sheep across Europe. Serological diagnosis of EAE is routinely carried out by the complement fixation test, although the interpretation of results can often be difficult because of cross reaction with Chlamydophila pecorum, which also commonly infects sheep. The purpose of this study was to evaluate and compare four ELISAs developed at Moredun Research Institute and based on whole C. abortus elementary bodies (EBs), an outer membrane preparation of the whole organism (SolPr) and two recombinant polymorphic outer membrane protein fragments (rOMP90-3 and rOMP90-4), with 3 commercial tests, the CHEKIT Chlamydophila Abortus, Pourquier ELISA Chlamydophila abortus and ImmunoComb Ovine Chlamydophila Antibody tests. The tests were evaluated using a panel of 202 sera from experimentally and naturally infected animals, as well as from EAE-free flocks. The EB, SolPr and CHEKIT ELISAs performed similarly to the CFT, all lacking in specificity by cross reacting with sera from C. pecorum infected animals. The ImmunoComb also lacked specificity with C. pecorum sera, but also badly cross reacted with sera from EAE-free flocks. The rOMP90-3, rOMP90-4 and Pourquier ELISAs were the most specific, although the Pourquier test appeared less sensitive with sera from naturally infected animals. Overall, the rOMP90-3 ELISA performed the best, with high sensitivity (96.8%) and no cross reaction with sera from C. pecorum infected animals or from EAE-free flocks (100% specificity) and so would be a suitable alternative to the CFT for the serological diagnosis of EAE.

  8. Production of a novel multi-epitope vaccine based on outer membrane proteins of Klebsiella pneumoniae

    Directory of Open Access Journals (Sweden)

    Tayebeh Farhadi

    2015-09-01

    Full Text Available Klebsiella pneumoniae is a hospital-acquired pathogen that leads to various infections. Hence, efforts to develop an effective vaccine against that pathogen are well documented. Our interest is the production of the previously designed multi-epitope vaccine construct against the K. pneumoniae in a prokaryotic host. Therefore, a new construct containing the nucleotide sequence of the novel vaccine was successfully expressed in Escherichia coli and then purified by Ni-NTA spin column. The purified recombinant protein can be considered as potential vaccine candidate for wet-laboratory analysis aiming to fight K pneumoniae.

  9. Klebsiella pneumoniae outer membrane protein A is required to prevent the activation of airway epithelial cells.

    Science.gov (United States)

    March, Catalina; Moranta, David; Regueiro, Verónica; Llobet, Enrique; Tomás, Anna; Garmendia, Junkal; Bengoechea, José A

    2011-03-25

    Outer membrane protein A (OmpA) is a class of proteins highly conserved among the Enterobacteriaceae family and throughout evolution. Klebsiella pneumoniae is a capsulated gram-negative pathogen. It is an important cause of community-acquired and nosocomial pneumonia. Evidence indicates that K. pneumoniae infections are characterized by a lack of an early inflammatory response. Data from our laboratory indicate that K. pneumoniae CPS helps to suppress the host inflammatory response. However, it is unknown whether K. pneumoniae employs additional factors to modulate host inflammatory responses. Here, we report that K. pneumoniae OmpA is important for immune evasion in vitro and in vivo. Infection of A549 and normal human bronchial cells with 52OmpA2, an ompA mutant, increased the levels of IL-8. 52145-Δwca(K2)ompA, which does not express CPS and ompA, induced the highest levels of IL-8. Both mutants could be complemented. In vivo, 52OmpA2 induced higher levels of tnfα, kc, and il6 than the wild type. ompA mutants activated NF-κB, and the phosphorylation of p38, p44/42, and JNK MAPKs and IL-8 induction was via NF-κB-dependent and p38- and p44/42-dependent pathways. 52OmpA2 engaged TLR2 and -4 to activate NF-κB, whereas 52145-Δwca(K2)ompA activated not only TLR2 and TLR4 but also NOD1. Finally, we demonstrate that the ompA mutant is attenuated in the pneumonia mouse model. The results of this study indicate that K. pneumoniae OmpA contributes to attenuate airway cell responses. This may facilitate pathogen survival in the hostile environment of the lung.

  10. Proteolytic cleavage of the Chlamydia pneumoniae major outer membrane protein in the absence of Pmp10

    DEFF Research Database (Denmark)

    Juul, Nicolai Stefan; Timmerman, E; Gevaert, K;

    2007-01-01

    that Pmp10 is differentially expressed in the C. pneumoniae CWL029 isolate. To evaluate whether the absence of Pmp10 in the outer membrane causes further changes to the C. pneumoniae protein profile, we subcloned the CWL029 isolate and selected a clone with minimal Pmp10 expression. Subsequently, we...... compared the proteome of the CWL029 isolate with the proteome of the subcloned strain and identified a specific cleavage of the C-terminal part of the major outer membrane protein (MOMP), which occurred only in the absence of Pmp10. In contrast, when Pmp10 was expressed we predominantly observed full...

  11. Usefulness of C-reactive protein in monitoring the severe community-acquired pneumonia clinical course.

    Science.gov (United States)

    Coelho, Luís; Póvoa, Pedro; Almeida, Eduardo; Fernandes, Antero; Mealha, Rui; Moreira, Pedro; Sabino, Henrique

    2007-01-01

    The aim of the present study was to evaluate the C-reactive protein level, the body temperature and the white cell count in patients after prescription of antibiotics in order to describe the clinical resolution of severe community-acquired pneumonia. A cohort of 53 consecutive patients with severe community-acquired pneumonia was studied. The C-reactive protein levels, body temperature and white cell count were monitored daily. By day 3 a C-reactive protein level 0.5 times the initial level was a marker of poor outcome (sensitivity, 0.91; specificity, 0.59). Patients were divided according to their C-reactive protein patterns of response to antibiotics, into fast response, slow response, nonresponse, and biphasic response. About 96% of patients with a C-reactive protein pattern of fast response and 74% of patients with a slow response pattern survived, whereas those patients with the patterns of nonresponse and of biphasic response had a mortality rate of 100% and 33%, respectively (P prescription is useful in identification, as early as day 3, of severe community-acquired pneumonia patients with poor outcome. The identification of the C-reactive protein pattern of response to antibiotic therapy was useful in the recognition of the individual clinical course, either improving or worsening, as well as the rate of improvement, in patients with severe community-acquired pneumonia.

  12. Production and Identification of Monoclonal Antibodies against MIP Protein of Chlamydophila abortus%流产嗜性衣原体MIP蛋白单克隆抗体的制备及特性鉴定

    Institute of Scientific and Technical Information of China (English)

    蔺俊丽; 李兆才; 曹小安; 赵荣; 周继章

    2015-01-01

    In order to produce the monoclonal antibodies ( mAbs) against MIP protein of Chlamydophila abortus, Balb/c mice were immunized with purified MIP protein. The positive hybridoma cell lines were screened by indirect ELISA after cell fusion and subcloned by limiting dilution. Two monoclonal antibody hybridoma cell lines which secrete MIP resistant protein were established eventually and named M1 and M3 respectively. The titer of these mAbs were1 ∶ 1600 and the relative affinity were 10 μg/mL by the way of ELISA;the recombinant MIP protein specificity of two strains of the monoclonal was recognized by Western Blot; the characteristics of hybridoma was confirmed by chromosome identification. The subtype of IgG of two strains was IgG1, and the light chain of them was κ chain by the mAb subclass detection kit; the neutralization test showed that the mAbs can interrupt transmission of infection. The successful preparation of the 2 mAbs anti-MIP will provide some useful date for setting up diagnostic and therapy method and the basic research of MIP protein in the future.%为制备抗流产嗜性衣原体MIP 蛋白单克隆抗体,以纯化的重组MIP 蛋白为抗原,免疫Balb/c小鼠后经细胞融合,间接ELISA法筛选阳性细胞克隆并进行有限稀释克隆化,建立了两株分泌抗MIP蛋白单克隆抗体的杂交瘤细胞株,将其分别命名为M1,M2。间接ELISA测杂交瘤细胞上清抗体效价为1∶1600、相对亲和力为10μg/mL;Western blot鉴定两株单克隆抗体能特异识别MIP重组蛋白;细胞核型实验鉴定单抗符合杂交瘤细胞的特性;单克隆抗体亚类检测试剂盒鉴定两株单抗免疫球蛋白类型均为IgG1,轻链为κ链;体外中和试验表明,两株单抗均能有效阻断感染。抗MIP单克隆抗体的成功制备将为建立诊断、治疗方法及MIP蛋白的进一步研究奠定基础。

  13. Cloning and Expression of Major Outer Membrane Protein of Chlamydophila Clinical Strain HB1043 from Swine%猪源嗜性衣原体HB1043株主要外膜蛋白(MOMP)基因的分析及重组表达

    Institute of Scientific and Technical Information of China (English)

    闫少侠; 田永祥; 刘泽文; 袁芳艳; 郭锐; 段正赢; 杨克礼; 孟丽; 周丹娜

    2013-01-01

    Chlamydophila was an obligate intracecullar parasites and could infect human and multiple animals.According to the published complete nucleotide sequence of Chlamydophila major outer membrane protein (MOMP) in the GenBank and the whole genome of Chlamydophila isolated from clinical samples,the whole genome sequence of MOMP gene were obtained by PCR.The PCR product was sequenced and blasted with the committed MOMP sequence (EU531729) from strain CG1 of swine Chiamydophila abortus.The results showed that it had one base different from the CG1 strain.The extinction enzymes sites BamH Ⅰ and Sol Ⅰ were selected for cloning the MOMP sequence to the pET-28a.The restructuring plasmid pET-28α-MOMP were then transfered into E.coli BL21 and expressed.The target protein rMOMP was purified by the Ni Sepharose 6 Fast Flow and proved with immunocompetence by Western-blot.%根据GenBank中衣原体主要外膜蛋白(MOMP)全基因序列设计特异性引物,以临床分离的衣原体病原中提取的衣原体全基因组为模板,利用特异性引物PCR扩增得到MOMP全基因序列.通过BLAST比对,结果显示其与已提交的猪流产衣原体CG1株的MOMP序列(EU531729)有1个碱基不同.根据对测序结果进行信号肽序列预测及内切酶位点分析,选用BamH Ⅰ和Sal Ⅰ双酶切将目的蛋白序列克隆到表达载体pET-28a上,得到重组质粒pET-28a-MOMP.将重组质粒转化入E coli BL21进行表达,表达产物使用镍离子亲和层析柱进行纯化得到目的蛋白rMOMP.目的蛋白经Western-blot检测,证明其具有免疫学活性.

  14. A Compendium for Mycoplasma pneumoniae.

    Science.gov (United States)

    Parrott, Gretchen L; Kinjo, Takeshi; Fujita, Jiro

    2016-01-01

    Historically, atypical pneumonia was a term used to describe an unusual presentation of pneumonia. Currently, it is used to describe the multitude of symptoms juxtaposing the classic symptoms found in cases of pneumococcal pneumonia. Specifically, atypical pneumonia is a syndrome resulting from a relatively common group of pathogens including Chlamydophila sp., and Mycoplasma pneumoniae. The incidence of M. pneumoniae pneumonia in adults is less than the burden experienced by children. Transmission rates among families indicate children may act as a reservoir and maintain contagiousness over a long period of time ranging from months to years. In adults, M. pneumoniae typically produces a mild, "walking" pneumonia and is considered to be one of the causes of persistent cough in patients. M. pneumoniae has also been shown to trigger the exacerbation of other lung diseases. It has been repeatedly detected in patients with bronchitis, asthma, chronic obstructive pulmonary disorder, and cystic fibrosis. Recent advances in technology allow for the rapid diagnosis of M. pneumoniae through the use of polymerase chain reaction or rapid antigen tests. With this, more effort has been afforded to identify the causative etiologic agent in all cases of pneumonia. However, previous practices, including the overprescribing of macrolide treatment in China and Japan, have created increased incidence of macrolide-resistant M. pneumoniae. Reports from these countries indicate that >85% of M. pneumoniae pneumonia pediatric cases are macrolide-resistant. Despite its extensively studied past, the smallest bacterial species still inspires some of the largest questions. The developments in microbiology, diagnostic features and techniques, epidemiology, treatment and vaccines, and upper respiratory conditions associated with M. pneumoniae in adult populations are included within this review.

  15. A compendium for Mycoplasma pneumoniae

    Directory of Open Access Journals (Sweden)

    Gretchen Lynn Parrott

    2016-04-01

    Full Text Available Historically, atypical pneumonia was a term used to describe an unusual presentation of pneumonia. Currently, it is used to describe the multitude of symptoms juxtaposing the classic symptoms found in cases of pneumococcal pneumonia. Specifically, atypical pneumonia is a syndrome resulting from a relatively common group of pathogens including Chlamydophila sp., and Mycoplasma pneumoniae. The incidence of M. pneumoniae pneumonia in adults is less than the burden experienced by children. Transmission rates among families indicate children may act as a reservoir and maintain contagiousness over a long period of time ranging from months to years. In adults, M. pneumoniae typically produces a mild, walking pneumonia and is considered to be one of the causes of persistent cough in patients. M. pneumoniae has also been shown to trigger the exacerbation of other lung diseases. It has been repeatedly detected in patients with bronchitis, asthma, chronic obstructive pulmonary disorder and cystic fibrosis. Recent advances in technology allow for the rapid diagnosis of M. pneumoniae through the use of polymerase chain reaction (PCR or rapid antigen tests. With this, more effort has been afforded to identify the causative etiologic agent in all cases of pneumonia. However, previous practices, including the overprescribing of macrolide treatment in China and Japan, have created increased incidence of macrolide-resistant M. pneumoniae. Reports from these countries indicate that >85% of M. pneumoniae pneumonia pediatric cases are macrolide-resistant. Despite its extensively studied past, the smallest bacterial species still inspires some of the largest questions. The developments in microbiology, diagnostic features and techniques, epidemiology, treatment and vaccines, and upper respiratory conditions associated with M. pneumoniae in adult populations are included within this review.

  16. Detection of Chlamydophila abortus in Sheep (Ovis aries in Mexico

    Directory of Open Access Journals (Sweden)

    Juan M. Jiménez-Estrada

    2008-01-01

    Full Text Available Chlamydophila abortus is one of the pathogens which induce abortion in small ruminants; this pathogen has a tropism for ruminant placenta and causes the disease commonly referred to as Ovine Enzootic Abortion (OEA. In Europe are estimated economic losses of around 20 million pounds a year by OEA. In the American Continent the disease has been reported only in Canada, the United States, Colombia and Chile while in Mexico it is unknown whether OEA is common and it is causing abortions in flocks of sheep from “Estado de Mexico”. The objective of this study was investigating the prevalence of anti-Chlamydophila abortus IgG antibodies and detection of C. abortus DNA in sheep with clinical abort history by mean of ELISA assay (C. abortus ELISA, Institute Pourquier, Montpellier, France and molecular identification of the principal outer membrane protein (POMP 90-91B gene by PCR, respectively. A cross-sectional study was carried out to enroll and random sample of ewes from november 2003 until march 2005. A total of 349 sera and vaginal swabs samples were collected from 35 flocks of sheep from Xalatlaco. The results showed that the seropositive rate was 31.1% (14/45 for healthy and 21.3% (65/304 for sheep with history clinical of abort. In vaginal swabs, the PCR showed 0% (0/45 for healthy animals and 0.65% (2/304 for aborted sheep. Samples from the lungs and liver of the fetus of one of these animals were also positive for C. abortus. In conclusion, these results confirmed that infection with C. abortus is common and is affecting sheep flocks in the Mexican highlands. Therefore, is necessary that the authorities responsible for animal welfare in Mexico (SAGARPA to set up appropriate epidemiological surveillance and control programs to eradicate this disease.

  17. Evaluation of two commercial assays for the detection of Chlamydophila abortus antibodies.

    Science.gov (United States)

    Vretou, E; Radouani, F; Psarrou, E; Kritikos, I; Xylouri, E; Mangana, O

    2007-07-20

    Two commercial enzyme-linked immunosorbent assays (ELISA), the CHEKIT-CHLAMYDIA which uses inactivated Chlamydophila psittaci antigen, and the Chlamydophila abortus ELISA produced by the Institut Pourquier which uses a recombinant fragment of the 80-90 kDa protein, were evaluated with the objective to determine whether the new ELISAs would perform as improved alternatives to the complement fixation test (CFT) for the serological diagnosis of ovine enzootic abortion (OEA). The results were compared to those obtained by the CFT and the competitive ELISA (cELISA). The tests were assessed with a panel of 17 serum samples from specific pathogen-free (SPF) lambs experimentally infected with various subtypes of Chlamydophila pecorum, with sera from 45 C. abortus-infected pregnant sheep and from 54 sheep free of OEA. The C. abortus ELISA was identified as being more specific and sensitive than the other tests. The 4 assays were evaluated further with 254 sera from flocks with documented OEA, from flocks with no history of abortion and from animals after abortion of unknown cause. The C. abortus ELISA by the Institut Pourquier identified less OEA-positive sera than the other assays though it identified correctly 9 of 10 OEA-positive flocks. The basis of the discordant results is discussed.

  18. Genus delineation of Chlamydiales by analysis of the percentage of conserved proteins justifies the reunifying of the genera Chlamydia and Chlamydophila into one single genus Chlamydia.

    Science.gov (United States)

    Pannekoek, Yvonne; Qi-Long, Qin; Zhang, Yu-Zhong; van der Ende, Arie

    2016-08-01

    Many studies have aimed to set up boundaries for the classification and definition of prokaryotic genus and species classification; however, studies that focused on genus-level genomic differences for existing taxonomy systems are limited. Recently, a novel method was described for prokaryotic genus delineation using the percentage of conserved proteins (POCP) between two strains to estimate their evolutionary and phenotypic distance (Qin et al. A proposed genus boundary for the prokaryotes based on genomic insights. J Bacteriol 2014; 196: :2210-5). Here, we extended the POCP analysis of the order Chlamydiales and pairwise compared all currently recognized species and candidate species of the family Chlamydiaceae as well as some species from other families. Using the taxonomy advised by the International Committee on Systematics of Prokaryotes, subcommittee on the taxonomy of the Chlamydiae, POCP analysis revealed that all pairwise comparisons of species from different families resulted in values lower than 50%, the proposed threshold for genus boundary. In contrast, all interspecies pairwise comparisons of species from the single genus within the family Chlamydiaceae resulted in POCP values higher than 70%. We conclude that the recommended genus classification of the family Chlamydiaceae is rational and that POCP analyses can provide a robust genomic index for the taxonomy of members of the order Chlamydiales in terms of genus demarcation.

  19. The putative proteinase maturation protein A of Streptococcus pneumoniae is a conserved surface protein with potential to elicit protective immune responses

    NARCIS (Netherlands)

    K. Overweg (Karin); A. Kerr; M. Sluijter (Marcel); M.H. Jackson; T.J. Mitchell; A.P. de Jong; R. de Groot (Ronald); P.W.M. Hermans (Peter)

    2000-01-01

    textabstractSurface-exposed proteins often play an important role in the interaction between pathogenic bacteria and their host. We isolated a pool of hydrophobic, surface-associated proteins of Streptococcus pneumoniae. The opsonophagocytic activity of hyperimmune

  20. Immunization with LytB protein of Streptococcus pneumoniae activates complement-mediated phagocytosis and induces protection against pneumonia and sepsis.

    Science.gov (United States)

    Corsini, Bruno; Aguinagalde, Leire; Ruiz, Susana; Domenech, Mirian; Antequera, María Luisa; Fenoll, Asunción; García, Pedro; García, Ernesto; Yuste, Jose

    2016-12-07

    The cell wall glucosaminidase LytB of Streptococcus pneumoniae is a surface exposed protein involved in daughter cell separation, biofilm formation and contributes to different aspects of the pathogenesis process. In this study we have characterized the antibody responses after immunization of mice with LytB in the presence of alhydrogel as an adjuvant. Enzyme-linked immunosorbent assays measuring different subclasses of immunoglobulin G, demonstrated that the antibody responses to LytB were predominantly IgG1 and IgG2b, followed by IgG3 and IgG2a subclasses. Complement-mediated immunity against two different pneumococcal serotypes was investigated using sera from immunized mice. Immunization with LytB increased the recognition of S. pneumoniae by complement components C1q and C3b demonstrating that anti-LytB antibodies trigger activation of the classical pathway. Phagocytosis assays showed that serum containing antibodies to LytB stimulates neutrophil-mediated phagocytosis against S. pneumoniae. Animal models of infection including invasive pneumonia and sepsis were performed with two different clinical isolates. Vaccination with LytB increased bacterial clearance and induced protection demonstrating that LytB might be a good candidate to be considered in a future protein-based vaccine against S. pneumoniae.

  1. Epizootic abortion related to infections by Chlamydophila abortus and Chlamydophila pecorum in water buffalo (Bubalus bubalis).

    Science.gov (United States)

    Greco, G; Corrente, M; Buonavoglia, D; Campanile, G; Di Palo, R; Martella, V; Bellacicco, A L; D'Abramo, M; Buonavoglia, C

    2008-06-01

    Water buffalo (Bubalus bubalis) are affected by high rates of embryonic mortality and abortion related to infectious diseases and non-infectious factors. A number of viral and bacterial infections have been associated with reproductive failure, but there is limited information on the role of chlamydial infections. In order to investigate the presence and the role of Chlamydiaceae in water buffalo a retrospective study was performed in a herd with a history of reproductive failure. During an 11-month period, the pregnant heifers suffered an abortion rate of 36.8% between the 3rd and 7th month of pregnancy. Antibodies to Chlamydiaceae were detected in 57% of the aborted cows, and in 0% of the overtly healthy cows used as control. By a nested-PCR assay, three of 14 vaginal swabs from aborted animals tested positive for Chlamydophila agents and, additionally, three out of seven aborted fetuses tested positive for Chlamydophila spp., with two being co-infections by Cp. abortus and Cp. pecorum and one being characterised as Cp. abortus. Sequence analysis of the amplicons confirmed the results of the nested-PCR. The presence of anti-Chlamydiaceae antibodies in more than half of the aborting animals (PChlamydophila agents in several fetal organs and in the vaginal swabs are consistent with the history of abortions observed in the herd and suggest an abortifacient role by Chlamydophila spp. in water buffalo (B. Bubalis) herds.

  2. Characterization of Spbhp-37, a Hemoglobin-Binding Protein of Streptococcus pneumoniae

    Science.gov (United States)

    Romero-Espejel, María E.; Rodríguez, Mario A.; Chávez-Munguía, Bibiana; Ríos-Castro, Emmanuel; Olivares-Trejo, José de Jesús

    2016-01-01

    Streptococcus pneumoniae is a Gram-positive microorganism that is the cause of bacterial pneumonia, sinusitis and otitis media. This human pathogen also can cause invasive diseases such as meningitis, bacteremia and septicemia. Hemoglobin (Hb) and haem can support the growth and viability of S. pneumoniae as sole iron sources. Unfortunately, the acquisition mechanism of Hb and haem in this bacterium has been poorly studied. Previously we identified two proteins of 37 and 22 kDa as putative Hb- and haem-binding proteins (Spbhp-37 and Spbhp-22, respectively). The sequence of Spbhp-37 protein was database annotated as lipoprotein without any function or localization. Here it was immunolocalized in the surface cell by transmission electron microscopy using specific antibodies produced against the recombinant protein. The expression of Spbhp-37 was increased when bacteria were grown in media culture supplied with Hb. In addition, the affinity of Sphbp-37 for Hb was determined. Thus, in this work we are presenting new findings that attempt to explain the mechanism involved in iron acquisition of this pathogen. In the future these results could help to develop new therapy targets in order to avoid the secondary effects caused by the traditional therapies. PMID:27200302

  3. Characterization of Spbhp-37, a Hemoglobin-Binding Protein of Streptococcus pneumoniae.

    Science.gov (United States)

    Romero-Espejel, María E; Rodríguez, Mario A; Chávez-Munguía, Bibiana; Ríos-Castro, Emmanuel; Olivares-Trejo, José de Jesús

    2016-01-01

    Streptococcus pneumoniae is a Gram-positive microorganism that is the cause of bacterial pneumonia, sinusitis and otitis media. This human pathogen also can cause invasive diseases such as meningitis, bacteremia and septicemia. Hemoglobin (Hb) and haem can support the growth and viability of S. pneumoniae as sole iron sources. Unfortunately, the acquisition mechanism of Hb and haem in this bacterium has been poorly studied. Previously we identified two proteins of 37 and 22 kDa as putative Hb- and haem-binding proteins (Spbhp-37 and Spbhp-22, respectively). The sequence of Spbhp-37 protein was database annotated as lipoprotein without any function or localization. Here it was immunolocalized in the surface cell by transmission electron microscopy using specific antibodies produced against the recombinant protein. The expression of Spbhp-37 was increased when bacteria were grown in media culture supplied with Hb. In addition, the affinity of Sphbp-37 for Hb was determined. Thus, in this work we are presenting new findings that attempt to explain the mechanism involved in iron acquisition of this pathogen. In the future these results could help to develop new therapy targets in order to avoid the secondary effects caused by the traditional therapies.

  4. Characterization of Spbhp-37, a haemoglobin-binding protein of Streptococcus pneumoniae

    Directory of Open Access Journals (Sweden)

    María Elena eRomero-Espejel

    2016-05-01

    Full Text Available Streptococcus pneumoniae is a Gram-positive microorganism that is the cause of bacterial pneumonia, sinusitis and otitis media. This human pathogen also can cause invasive diseases such as meningitis, bacteremia and septicemia. Haemoglobin (Hb and haem can support the growth and viability of S. pneumoniae as sole iron sources. Unfortunately, the acquisition mechanism of Hb and haem in this bacterium has been poorly studied. Previously we identified two proteins of 37 and 22 kDa as putative Hb- and haem-binding proteins (Spbhp-37 and Spbhp-22, respectively. The sequence of Spbhp-37 protein was database annotated as lipoprotein without any function or localization. Here it was immunolocalized in the surface cell by transmission electron microscopy using specific antibodies produced against the recombinant protein. The expression of Spbhp-37 was increased when bacteria were grown in media culture supplied with Hb. In addition, the affinity of Sphbp-37 for Hb was determined. Thus, in this work we are presenting new findings that attempt to explain the mechanism involved in iron acquisition of this pathogen. In the future these results could help to develop new therapy targets in order to avoid the secondary effects caused by the traditional therapies.

  5. NCBI nr-aa BLAST: CBRC-CPOR-01-0641 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CPOR-01-0641 ref|NP_224724.1| Glutamate Symport [Chlamydophila pneumoniae CWL0...29] ref|NP_300583.1| glutamate symport [Chlamydophila pneumoniae J138] ref|NP_444775.1| sodium:dicarboxylate... symporter family protein [Chlamydophila pneumoniae AR39] ref|NP_876821.1| proton/sodium glutamate symport transporter [Chlamydophila... pneumoniae TW-183] gb|AAD18668.1| Glutamate Symport [Chlamydophila... pneumoniae CWL029] gb|AAF73644.1| sodium:dicarboxylate symporter family protein [Chlamydophila p

  6. Chlamydophila felis in cats--are the stray cats dangerous source of infection?

    Science.gov (United States)

    Halánová, M; Sulinová, Z; Cisláková, L; Trbolová, A; Páleník, L; Weissová, T; Halán, M; Kalinová, Z; Holičková, M

    2011-11-01

    Chlamydophila felis is a causative agent of acute or chronic conjunctivitis, and pneumonia in cats. Natural transmission mostly occurs consequently to close contact with other infected cats, their aerosol and fomites. We have examined 93 cats with symptoms of acute or chronic conjunctivitis, from Košice region in Slovakia, during the period of 2 years. Conjunctival samples were obtained from 55 domestic cats (59.14%) and 38 stray cats (40.86%). Of the total number of 93 examined animals, 42 cats were positive, which represents 45.16% overall positivity. Out of the 42 positive cats, 25 cats were stray and 17 positive cats were classified as domestic, which means that of 38 stray cats, 25 were positive (which represented 65.78% positivity) and of 55 domestic cats, 17 were positive (positivity was 30.90%). Our results showed that cats, especially stray cats, could be a dangerous source of chlamydiosis for humans.

  7. Colistin resistance associated with outer membrane protein change in Klebsiella pneumoniae and Enterobacter asburiae.

    Science.gov (United States)

    Kádár, Béla; Kocsis, Béla; Tóth, Ákos; Kristóf, Katalin; Felső, Péter; Kocsis, Béla; Böddi, Katalin; Szabó, Dóra

    2017-06-01

    In this study, outer membrane proteins (OMPs) of colistin-resistant Klebsiella pneumoniae and Enterobacter asburiae were analyzed. One colistin-susceptible and three colistin-resistant K. pneumoniae sequence type 258 strains as well as one colistin-susceptible E. asburiae and its colistin-heteroresistant counterpart strain were involved in the study. OMP analysis of each strain was performed by microchip method. Matrix-assisted laser desorption ionization time of flight/mass spectrometry (MALDI-TOF/MS) investigation was carried out after separation of OMPs by two-dimensional gel electrophoresis and in-gel digestion. The MALDI-TOF/MS analysis of OMPs in the colistin-susceptible K. pneumoniae found 16 kDa proteins belonging to the LysM domain/BON superfamily, as well as DNA starvation proteins, whereas OmpX and OmpW were detected in the colistin-resistant counterpart strains. OmpC and OmpW were detected in the colistin-susceptible E. asburiae, whereas OmpA and OmpX were identified in the colistin-resistant counterpart. This study demonstrated that OMP differences were between colistin-susceptible and -resistant counterpart strains. The altered Gram-negative cell wall may contribute to acquired colistin resistance in Enterobacteriaceae.

  8. Roles of the Essential Protein FtsA in Cell Growth and Division in Streptococcus pneumoniae

    Science.gov (United States)

    Fadda, Daniela; Perez, Amilcar J.; Danforth, Madeline L.; Musu, Daniela; Krupka, Marcin; Denapaite, Dalia; Tsui, Ho-Ching T.; Winkler, Malcolm E.; Branny, Pavel

    2016-01-01

    ABSTRACT Streptococcus pneumoniae is an ovoid-shaped Gram-positive bacterium that grows by carrying out peripheral and septal peptidoglycan (PG) synthesis, analogous to model bacilli, such as Escherichia coli and Bacillus subtilis. In the model bacilli, FtsZ and FtsA proteins assemble into a ring at midcell and are dedicated to septal PG synthesis but not peripheral PG synthesis; hence, inactivation of FtsZ or FtsA results in long filamentous cells unable to divide. Here, we demonstrate that FtsA and FtsZ colocalize at midcell in S. pneumoniae and that partial depletion of FtsA perturbs septum synthesis, resulting in elongated cells with multiple FtsZ rings that fail to complete septation. Unexpectedly, complete depletion of FtsA resulted in the delocalization of FtsZ rings and ultimately cell ballooning and lysis. In contrast, depletion or deletion of gpsB and sepF, which in B. subtilis are synthetically lethal with ftsA, resulted in enlarged and elongated cells with multiple FtsZ rings, with deletion of sepF mimicking partial depletion of FtsA. Notably, cell ballooning was not observed, consistent with later recruitment of these proteins to midcell after Z-ring assembly. The overproduction of FtsA stimulates septation and suppresses the cell division defects caused by the deletion of sepF and gpsB under some conditions, supporting the notion that FtsA shares overlapping functions with GpsB and SepF at later steps in the division process. Our results indicate that, in S. pneumoniae, both GpsB and SepF are involved in septal PG synthesis, whereas FtsA and FtsZ coordinate both peripheral and septal PG synthesis and are codependent for localization at midcell. IMPORTANCE Streptococcus pneumoniae (pneumococcus) is a clinically important human pathogen for which more therapies against unexploited essential targets, like cell growth and division proteins, are needed. Pneumococcus is an ovoid-shaped Gram-positive bacterium with cell growth and division properties that

  9. Systems Biology Approaches for the Prediction of Possible Role of Chlamydia pneumoniae Proteins in the Etiology of Lung Cancer.

    Science.gov (United States)

    Khan, Shahanavaj; Imran, Ahamad; Khan, Abdul Arif; Abul Kalam, Mohd; Alshamsan, Aws

    2016-01-01

    Accumulating evidence has recently supported the association of bacterial infection with the growth and development of cancers, particularly in organs that are constantly exposed to bacteria such as the lungs, colon, cervical cancer etc. Our in silico study on the proteome of Chlamydia pneumoniae suggests an unprecedented idea of the etiology of lung cancer and have revealed that the infection of C. pneumoniae is associated with lung cancer development and growth. It is reasonable to assume that C. pneumoniae transports its proteins within host-intracellular organelles during infection, where they may work with host-cell proteome. The current study was performed for the prediction of nuclear targeting protein of C. pneumoniae in the host cell using bioinformatics predictors including ExPASy pI/Mw tool, nuclear localization signal (NLS) mapper, balanced sub cellular localization predictor (BaCeILo), and Hum-mPLoc 2.0. We predicted 47/1112 nuclear-targeting proteins of C. pneumoniae connected with several possible alterations in host replication and transcription during intracellular infection. These nuclear-targeting proteins may direct to competitive interactions of host and C. pneumoniae proteins with the availability of same substrate and may be involved as etiological agents in the growth and development of lung cancer. These novel findings are expected to access in better understanding of lung cancer etiology and identifying molecular targets for therapy.

  10. Identification of proteins in Streptococcus pneumoniae by reverse vaccinology and genetic diversity of these proteins in clinical isolates.

    Science.gov (United States)

    Argondizzo, Ana Paula Corrêa; da Mota, Fabio Faria; Pestana, Cristiane Pinheiro; Reis, Joice Neves; de Miranda, Antonio Basílio; Galler, Ricardo; Medeiros, Marco Alberto

    2015-02-01

    Streptococcus pneumoniae is a major cause of morbidity and mortality worldwide. Virulence-associated proteins common and conserved among all capsular types now represent the best strategy to combat pneumococcal infections. Our aim was to identify conserved targets in pneumococci that showed positive prediction for lipoprotein and extracellular subcellular location using bioinformatics programs and verify the distribution and the degree of conservation of these targets in pneumococci. These targets can be considered potential vaccine candidate to be evaluated in the future. A set of 13 targets were analyzed and confirmed the presence in all pneumococci tested. These 13 genes were highly conserved showing around >96 % of amino acid and nucleotide identity, but they were also present and show high identity in the closely related species Streptococcus mitis, Streptococcus oralis, and Streptococcus pseudopneumoniae. S. oralis clusters away from S. pneumoniae, while S. pseudopneumoniae and S. mitis cluster closer. The divergence between the selected targets was too small to be observed consistently in phylogenetic groups between the analyzed genomes of S. pneumoniae. The proteins analyzed fulfill two of the initial criteria of a vaccine candidate: targets are present in a variety of different pneumococci strains including different serotypes and are conserved among the samples evaluated.

  11. [Evaluation of penicillin-binding protein genotypes in penicillin susceptible and resistant Streptococcus pneumoniae isolates].

    Science.gov (United States)

    Aslan, Gönül; Tezcan, Seda; Delialioğlu, Nuran; Aydın, Fatma Esin; Kuyucu, Necdet; Emekdaş, Gürol

    2012-04-01

    Penicillin-binding proteins (PBPs) are the natural targets of beta-lactam antibiotics and mutations in pbp1a, pbp2b, and pbp2x genes, which encode PBPs, are responsible for resistance to beta-lactams in Streptococcus pneumoniae. In the present study, we intended to determine how often the common mutation patterns occurred within the pbp1a, pbp2b, and pbp2x PBP gene regions and evaluate the PBP genotype mutations which were associated with penicillin resistance in several penicillin-susceptible and - resistant S.pneumoniae isolates in Mersin, Turkey. A total of 62 S.pneumoniae strains isolated from different clinical specimens (32 nasopharyngeal swab, 16 sputum, 3 blood, 3 wound, 2 cerebrospinal fluids and one of each urine, abscess, bronchoalveolar lavage, conjunctival swab, tracheal aspirate, middle ear effusion) were included in the study. Penicillin susceptibilities of the isolates were searched by disc diffusion and E-test methods, and 23 of them were identified as susceptible, 31 were intermediate susceptible, and eight were resistant to penicillin. A rapid DNA extraction procedure was performed for the isolation of nucleic acids from the strains. Distribution of PBP gene mutations in pbp1a, pbp2b, and pbp2x gene regions related to penicillin resistance was determined by using a wild-type specific polymerase chain reaction (PCR) based technique. PBP gene alterations of those isolates were also evaluated in relation to penicillin susceptibility and resistance patterns. Twenty two (95.7%) of 23 penicillin-susceptible S.pneumoniae isolates exhibited no mutation in the three PBP genes (pbp1a, pbp2x, and pbp 2b), while 1 (4.3%) of these harbored mutations in all of the three PBP genes. The penicillin-intermediate susceptible S.pneumoniae isolates exhibited various combinations of mutations. One (3.2%) of 31 penicillin-intermediate susceptible isolates exhibited no mutation in the three PBP genes, while 22 (71%) of them yielded mutations in all of the three PBP

  12. Overexpression, purification and crystallization of a choline-binding protein CbpI from Streptococcus pneumoniae

    Energy Technology Data Exchange (ETDEWEB)

    Paterson, Neil G., E-mail: neison@chem.gla.ac.uk; Riboldi-Tunicliffe, Alan [Department of Chemistry and WestCHEM, Glasgow Biomedical Research Centre (GBRC), University of Glasgow, 120 University Place, Glasgow G12 8TA,Scotland (United Kingdom); Mitchell, Timothy J. [Division of Infection and Immunity (IBLS), Glasgow Biomedical Research Centre (GBRC), University of Glasgow, 120 University Place, Glasgow G12 8TA,Scotland (United Kingdom); Isaacs, Neil W. [Department of Chemistry and WestCHEM, Glasgow Biomedical Research Centre (GBRC), University of Glasgow, 120 University Place, Glasgow G12 8TA,Scotland (United Kingdom)

    2006-07-01

    The choline-binding protein CbpI from S. pneumoniae has been purified and crystallized and diffraction data have been collected to 3.5 Å resolution. The choline-binding protein CbpI from Streptococcus pneumoniae is a 23.4 kDa protein with no known function. The protein has been successfully purified initially using Ni–NTA chromatography and to homogeneity using Q-Sepharose ion-exchange resin as an affinity column. CbpI was crystallized using PEG 3350 as a precipitant and X-ray crystallographic analysis showed that the crystals belonged to the tetragonal space group P4, with unit-cell parameters a = b = 83.31, c = 80.29 Å, α = β = γ = 90°. The crystal contains two molecules in the asymmetric unit with a solvent content of 55.7% (V{sub M} = 2.77 Å{sup 3} Da{sup −1}) and shows a diffraction limit of 3.5 Å.

  13. Role of Streptococcus pneumoniae Proteins in Evasion of Complement-Mediated Immunity

    Science.gov (United States)

    Andre, Greiciely O.; Converso, Thiago R.; Politano, Walter R.; Ferraz, Lucio F. C.; Ribeiro, Marcelo L.; Leite, Luciana C. C.; Darrieux, Michelle

    2017-01-01

    The complement system plays a central role in immune defense against Streptococcus pneumoniae. In order to evade complement attack, pneumococci have evolved a number of mechanisms that limit complement mediated opsonization and subsequent phagocytosis. This review focuses on the strategies employed by pneumococci to circumvent complement mediated immunity, both in vitro and in vivo. At last, since many of the proteins involved in interactions with complement components are vaccine candidates in different stages of validation, we explore the use of these antigens alone or in combination, as potential vaccine approaches that aim at elimination or drastic reduction in the ability of this bacterium to evade complement. PMID:28265264

  14. Chlamydophila abortus em animais de produção Chlamydophila abortus in production animals

    Directory of Open Access Journals (Sweden)

    Francielle Gibson da Silva

    2006-02-01

    Full Text Available A Chlamydophila abortus (anteriormente classificada como Chlamydia psittaci sorotipo 1 tem sido descrita em muitos países, associada principalmente com distúrbios reprodutivos em ovinos, bovinos e caprinos. O aborto enzoótico dos ovinos e caprinos e o aborto epizoótico dos bovinos são as doenças mais importantes causadas por esta bactéria. No Brasil, as pesquisas com C. abortus são praticamente inexistentes. O objetivo desta revisão é apresentar informações sobre modificações taxonômicas, ciclo de vida, epidemiologia, patogenia, sinais clínicos e diagnóstico da infecção por C. abortus principalmente em ovinos, bovinos e caprinos.Chlamydophila abortus (previously known as Chlamydia psittaci serovar 1 has been reported in many countries, associated with reproductive disorders in sheep, cattle, and goats. The enzootic abortion of sheep and goats and the epizootic bovine abortion are the most important diseases produced by this bacterium. In Brazil, there is scarce information about C. abortus. The objective of this review is to show information about taxonomic changes, life cycle, epidemiology, pathogenesis, clinical signs and diagnosis of C. abortus in sheep, cattle and goats.

  15. The Chlamydophila felis plasmid is highly conserved.

    Science.gov (United States)

    Harley, Ross; Day, Sarinder; Di Rocco, Camillo; Helps, Chris

    2010-11-20

    The presence of a plasmid in the Chlamydiaceae is both species and strain specific. Knowledge of the prevalence of the plasmid in different Chlamydia species is important for future studies aiming to investigate the role of the plasmid in chlamydial biology and disease. Although strains of Chlamydophila felis with or without the plasmid have been identified, only a small number of laboratory-adapted strains have been analysed and the prevalence of the plasmid in field isolates has not been determined. This study aimed to determine the prevalence of the plasmid in C. felis-positive conjunctival and oropharyngeal clinical samples submitted for routine diagnosis of C. felis by real-time (Q)PCR. DNA extracts from four laboratory-adapted strains were also analysed. QPCR assays targeting regions of C. felis plasmid genes pCF01, pCF02 and pCF03 were developed for the detection of plasmid DNA. QPCR analysis of DNA extracts from C. felis-positive clinical samples found evidence of plasmid DNA in 591 of 595 samples representing 561 of 564 (99.5%) clinical cases. Plasmid DNA was also detected by QPCR in laboratory-adapted strains 1497V, K2487 and K2490, but not strain 905. We conclude that the plasmid is highly conserved in C. felis, and plasmid-deficient strains represent a rare but important population for future studies of chlamydial plasmid function.

  16. Influence of ulinastatin on pulmonary surfactant protein, anti-inflammatory and pro-inflammatory mediator in patients with severe pneumonia

    Institute of Scientific and Technical Information of China (English)

    Li Wang; Rui Kang; Jia-Li Xie; Ya-Ni Xue

    2016-01-01

    Objective:To observe the influence of ulinastatin on pulmonary surfactant protein and anti-inflammatory and pro-inflammatory mediator in patients with severe pneumonia. Methods:A total of 54 patients with severe pneumonia treated in our hospital from April 2014 to May 2015 were selected as the study object, and they were randomly divided into control group (conventional treatment of severe pneumonia group) and observation group (conventional treatment and ulinastatin group), with 27 cases in each group. Then the serum levels of pulmonary surfactant protein,anti-inflammatory and pro-inflammatory mediators in two groups before and after treatment at 1 day, 3 day and 5 day were compared. Results:The serum level of pulmonary surfactant protein, anti-inflammatory and pro-inflammatory mediators in two groups before treatment had no significant differences, all P>0.05, and those serum indexes in observation group after treatment at 1 day, 3 day and 5 day were all significantly better than those of the control group, all P<0.05. Conclusions:The ulinastatin can effectively improve the pulmonary surfactant protein, anti-inflammatory and pro-inflammatory mediators in patients with severe pneumonia, and its improvement role for various of severe pneumonia are obvious.

  17. NCBI nr-aa BLAST: CBRC-DNOV-01-0217 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-0217 ref|NP_224546.1| Integral Membrane Protein [Chlamydophila pneumon...iae CWL029] ref|NP_300403.1| integral membrane protein [Chlamydophila pneumoniae J138] ref|NP_444963.1| ABC ...transporter, permease protein, putative [Chlamydophila pneumoniae AR39] ref|NP_876627.1| putative integral membrane protein [Chlamydo...etal transport system membrane protein CPn_0346/CP_0414/CPj0346/CpB0353 gb|AAD18490.1| Integral Membrane Protein [Chlamydophila... pneumoniae CWL029] gb|AAF38258.1| ABC transporter, permease protein, putative [Chlamydophila

  18. Recombinant human activated protein C inhibits local and systemic activation of coagulation without influencing inflammation during Pseudomonas aeruginosa pneumonia in rats

    NARCIS (Netherlands)

    Choi, Goda; Hofstra, Jorrit-Jan H; Roelofs, Joris J T H; Florquin, Sandrine; Bresser, Paul; Levi, Marcel; van der Poll, Tom; Schultz, Marcus J

    OBJECTIVE: Alveolar fibrin deposition is a hallmark of pneumonia. It has been proposed that recombinant human activated protein C exerts lung-protective effects via anticoagulant and anti-inflammatory pathways. We investigated the role of the protein C system in pneumonia caused by Pseudomonas

  19. Profiling of β-lactam selectivity for penicillin-binding proteins in Streptococcus pneumoniae D39.

    Science.gov (United States)

    Kocaoglu, Ozden; Tsui, Ho-Ching T; Winkler, Malcolm E; Carlson, Erin E

    2015-01-01

    Selective fluorescent β-lactam chemical probes enable the visualization of the transpeptidase activity of penicillin-binding proteins (PBPs) at different stages of bacterial cell division. To facilitate the development of new fluorescent probes for PBP imaging, we evaluated 20 commercially available β-lactams for selective PBP inhibition in an unencapsulated derivative of the D39 strain of Streptococcus pneumoniae. Live cells were treated with β-lactam antibiotics at different concentrations and subsequently incubated with Bocillin FL (Boc-FL; fluorescent penicillin) to saturate uninhibited PBPs. Fluorophore-labeled PBPs were visualized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and fluorescence scanning. Among 20 compounds tested, carbapenems (doripenem and meropenem) were coselective for PBP1a, PBP2x, and PBP3, while six of the nine penicillin compounds were coselective for PBP2x and PBP3. In contrast, the seven cephalosporin compounds tested display variability in their PBP-binding profiles. Three cephalosporin compounds (cefoxitin, cephalexin, and cefsulodin) and the monobactam aztreonam exhibited selectivity for PBP3, while only cefuroxime (a cephalosporin) was selective for PBP2x. Treatment of S. pneumoniae cultures with a sublethal concentration of cefuroxime that inhibited 60% of PBP2x activity and less than 20% of the activity of other PBPs resulted in formation of elongated cells. In contrast, treatment of S. pneumoniae cultures with concentrations of aztreonam and cefoxitin that inhibited up to 70% of PBP3 activity and less than 30% of other PBPs resulted in no discernible morphological changes. Additionally, correlation of the MIC and IC50s for each PBP, with the exception of faropenem, amdinocillin (mecillinam), and 6-APA, suggests that pneumococcal growth inhibition is primarily due to the inhibition of PBP2x.

  20. C reactive protein, calcitonin and D-dimer in patients of community acquired pneumonia

    Institute of Scientific and Technical Information of China (English)

    Xue-Lin Zhang; Zhen Wang; Shu-Hui Lv; Hai-Jun Jing; Jian-Yun Kang; Jian-Qing Zhao

    2016-01-01

    Objective:To investigate the clinical significance of C- reactive protein (CRP), calcitonin (PCT) and D- two (D-D) in community acquired pneumonia.Methods:A total of 102 patients with community-acquired pneumonia (CAP) admitted from March 2015- March 2016 as the research objects. A total of 5 mL peripheral venous blood of CAP patients (within 24 h of admission, before antibiotic therapy) were collected, and centrifuged to obtain serum. Immune turbidimetric method was used in determination of CRP and DD, immune fluorescence method was used for determination of PCT.Results:As grade increasing, the levels of CRP, PCT, D-D were increased gradually, with significant difference among different levels (P<0.05); CRP, PCT and D-D levels of severe group were significantly higher than those of non severe group (P<0.05); death group, CRP, PCT and D-D levels of death group were significantly higher than those of the survival group (P< 0.05).Conclusions:CRP, PCT, D-D levels have certain correlation with degree of severity. They can be used as important indicators to judge the severity of the disease, and predict the prognosis. High levels of CRP, PCT, D-D indicate severity of the disease and poor prognosis.

  1. cAMP receptor protein (CRP) downregulates Klebsiella pneumoniae nif promoters in Escherichia coli

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    In enteric bacteria, in response to the PTS system, the cAMP receptor protein (CRP) mediates the glucose effect, via regulating s70-dependent catabolic genes at transcriptional level. In this study, it is observed that the nitrogen fixation capacity of Klebsiella pneumoniae varies strongly when cells are grown on different carbohydrates, and this carbon effect occurs at the level of nif gene expression. Here we show that CRP can repress s54-dependent nif promoters (nifB, nifE, nifF, nifH, nifJ, nifLA and nifU), in a cAMP dependent fashion, in closed related E. coli background. Sequence analysis of these nif promoters indicates that there is no direct correlation between the fold of CRP-cAMP-mediated inhibition and the upstream cis elements at the promoters. In addition, the crp gene of K. pneumoniae has been isolated and sequenced, which is structural and functional highly homologous to that of E. coli. This suggests that CRP-cAMP-mediated inhibition on the nif promoters could be the reason for carbon effect on nitrogen fixation and thus has its physiological significance. A novel regulatory linkage between carbon metabolism and nitrogen fixation is proposed.

  2. cIAP-1 controls innate immunity to C. pneumoniae pulmonary infection.

    Directory of Open Access Journals (Sweden)

    Hridayesh Prakash

    Full Text Available The resistance of epithelial cells infected with Chlamydophila pneumoniae for apoptosis has been attributed to the induced expression and increased stability of anti-apoptotic proteins called inhibitor of apoptosis proteins (IAPs. The significance of cellular inhibitor of apoptosis protein-1 (cIAP-1 in C. pneumoniae pulmonary infection and innate immune response was investigated in cIAP-1 knockout (KO mice using a novel non-invasive intra-tracheal infection method. In contrast to wildtype, cIAP-1 knockout mice failed to clear the infection from their lungs. Wildtype mice responded to infection with a strong inflammatory response in the lung. In contrast, the recruitment of macrophages was reduced in cIAP-1 KO mice compared to wildtype mice. The concentration of Interferon gamma (IFN-gamma was increased whereas that of Tumor Necrosis Factor (TNF-alpha was reduced in the lungs of infected cIAP-1 KO mice compared to infected wildtype mice. Ex vivo experiments on mouse peritoneal macrophages and splenocytes revealed that cIAP-1 is required for innate immune responses of these cells. Our findings thus suggest a new immunoregulatory role of cIAP-1 in the course of bacterial infection.

  3. Decrease in penicillin susceptibility due to heat shock protein ClpL in Streptococcus pneumoniae.

    Science.gov (United States)

    Tran, Thao Dang-Hien; Kwon, Hyog-Young; Kim, Eun-Hye; Kim, Ki-Woo; Briles, David E; Pyo, Suhkneung; Rhee, Dong-Kwon

    2011-06-01

    Antibiotic resistance and tolerance are increasing threats to global health as antibiotic-resistant bacteria can cause severe morbidity and mortality and can increase treatment cost 10-fold. Although several genes contributing to antibiotic tolerance among pneumococci have been identified, we report here that ClpL, a major heat shock protein, could modulate cell wall biosynthetic enzymes and lead to decreased penicillin susceptibility. On capsular type 1, 2, and 19 genetic backgrounds, mutants lacking ClpL were more susceptible to penicillin and had thinner cell walls than the parental strains, whereas a ClpL-overexpressing strain showed a higher resistance to penicillin and a thicker cell wall. Although exposure of Streptococcus pneumoniae D39 to penicillin inhibited expression of the major cell wall synthesis gene pbp2x, heat shock induced a ClpL-dependent increase in the mRNA levels and protein synthesized by pbp2x. Inducible ClpL expression correlated with PBP2x expression and penicillin susceptibility. Fractionation and electron micrograph data revealed that ClpL induced by heat shock is localized at the cell wall, and the ΔclpL showed significantly reduced net translocation of PBP2x into the cell wall. Moreover, coimmunoprecipitation with either ClpL or PBP2x antibody followed by reprobing with ClpL or PBP2x antibody showed an interaction between ClpL and PBP2x after heat stress. This interaction was confirmed by His tag pulldown assay with either ClpLHis₆ or PBP2xHis₆. Thus, ClpL stabilized pbp2x expression, interacted with PBP2x, and facilitated translocation of PBP2x, a key protein of cell wall synthesis process, contributing to the decrease of antibiotic susceptibility in S. pneumoniae.

  4. Molecular epidemiology of C. pneumoniae infections

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    Alisa Shurdhi

    2010-06-01

    Full Text Available Introduction. Chlamydophila pneumoniae (C. pneumoniae is one of the most common respiratory pathogen, with an incidence of infection varying from 6% to 20%. The present study aimed to assess the incidence of C. pneumoniae infections in patients with acute respiratory diseases using a RealTime PCR (RT-PCR method. Methods. In the period January 2007-December 2008 279 biological samples coming from patients (190 males and 89 females with acute respiratory infections was collected and tested. Samples have been extracted using NucliSens easyMag Biomerieu according to manufacturer’s instructions and amplified by LightCycler Real-Time PCR Roche for the detection of C. pneumoniae DNA. Results. Data analysis revealed a higher prevalence of C. pneumoniae infections in male patients (7.9% than in females (5.6%. In addition, it is interesting to note that the incidence of C. pneumoniae infection was higher 28.6% in the period February - April. Conclusions. The results obtained in this study confirm the utility of molecular techniques in laboratory diagnosis and epidemiological investigations of respiratory infection caused by C. pneumoniae. RT-PCR have proved to be a rapid and a reliable technique to monitor and treat opportunely C. pneumoniae infections to avoid short and medium/long term complications.

  5. Study on the influence of puerarin injection for the pulmonary surfactant protein and inflammatory mediators of children with severe pneumonia

    Institute of Scientific and Technical Information of China (English)

    Lv-Wei Zhang

    2015-01-01

    Objective: To study and observe the influence situation of puerarin injection for the pulmonary surfactant protein and inflammatory mediators of children with severe pneumonia. Methods: 60 children with severe pneumonia in our hospital from February 2013 to January 2015 were selected as study object,and they were randomly divided into control group (routine treatment group) 30 cases and observation group (routine treatment and puerarin injection group) 30 cases, then the serum pulmonary surfactant protein and inflammatory mediators of two groups before the treatment and at different time after the treatment were respectively detected and compared. Results: The serum ulmonary surfactant protein and inflammatory mediators of observation group at third,fifth and tenth day after the treatment were all obviously lower than those of control group, all P<0.05, the comparison indexes after the treatment all had significant differences. Conclusions: The influence of puerarin injection for the pulmonary surfactant protein and inflammatory mediators of children with severe pneumonia are great, and it can effectively improve the disease state of children with severe pneumonia.

  6. The added value of C-reactive protein measurement in diagnosing pneumonia in primary care

    DEFF Research Database (Denmark)

    Minnaard, Margaretha C; de Groot, Joris A H; Hopstaken, Rogier M

    2017-01-01

    BACKGROUND: C-reactive protein (CRP) is increasingly being included in the diagnostic work-up for community-acquired pneumonia in primary care. Its added diagnostic value beyond signs and symptoms, however, remains unclear. We conducted a meta-analysis of individual patient data to quantify...... the added value of CRP measurement. METHODS: We included studies of the diagnostic accuracy of CRP in adult outpatients with suspected lower respiratory tract infection. We contacted authors of eligible studies for inclusion of data and for additional data as needed. The value of adding CRP measurement......, it still left a substantial group of patients classified at intermediate risk, in which clinical decision-making remains challenging....

  7. Comprehensive in silico prediction and analysis of chlamydial outer membrane proteins reflects evolution and life style of the Chlamydiae

    Directory of Open Access Journals (Sweden)

    Myers Garry

    2009-12-01

    Full Text Available Abstract Background Chlamydiae are obligate intracellular bacteria comprising some of the most important bacterial pathogens of animals and humans. Although chlamydial outer membrane proteins play a key role for attachment to and entry into host cells, only few have been described so far. We developed a comprehensive, multiphasic in silico approach, including the calculation of clusters of orthologues, to predict outer membrane proteins using conservative criteria. We tested this approach using Escherichia coli (positive control and Bacillus subtilis (negative control, and applied it to five chlamydial species; Chlamydia trachomatis, Chlamydia muridarum, Chlamydia (a.k.a. Chlamydophila pneumoniae, Chlamydia (a.k.a. Chlamydophila caviae, and Protochlamydia amoebophila. Results In total, 312 chlamydial outer membrane proteins and lipoproteins in 88 orthologous clusters were identified, including 238 proteins not previously recognized to be located in the outer membrane. Analysis of their taxonomic distribution revealed an evolutionary conservation among Chlamydiae, Verrucomicrobia, Lentisphaerae and Planctomycetes as well as lifestyle-dependent conservation of the chlamydial outer membrane protein composition. Conclusion This analysis suggested a correlation between the outer membrane protein composition and the host range of chlamydiae and revealed a common set of outer membrane proteins shared by these intracellular bacteria. The collection of predicted chlamydial outer membrane proteins is available at the online database pCOMP http://www.microbial-ecology.net/pcomp and might provide future guidance in the quest for anti-chlamydial vaccines.

  8. Aspiration pneumonia

    Science.gov (United States)

    Anaerobic pneumonia; Aspiration of vomitus; Necrotizing pneumonia; Aspiration pneumonitis ... The type of bacteria that caused the pneumonia depends on: Your ... facility, for example) Whether you were recently hospitalized ...

  9. Early Diagnosis of Pneumonia in Severe Stroke: Clinical Features and the Diagnostic Role of C-Reactive Protein.

    Directory of Open Access Journals (Sweden)

    Anushka Warusevitane

    Full Text Available Accurate diagnosis of pneumonia complicating severe stroke is challenging due to difficulties in physical examination, altered immune responses and delayed manifestations of radiological changes. The aims of this study were to describe early clinical features and to examine C-reactive protein (CRP as a diagnostic marker of post-stroke pneumonia.Patients who required nasogastric feeding and had no evidence of pneumonia within 7 days of stroke onset were included in the study and followed-up for 21 days with a daily clinical examination. Pneumonia was diagnosed using modified British Thoracic Society criteria.60 patients were recruited (mean age 77 years, mean National Institutes of Health Stroke Scale Score 19.47. Forty-four episodes of pneumonia were identified. Common manifestations on the day of the diagnosis were new onset crackles (43/44, 98%, tachypnoea>25/min (42/44, 95%, and oxygen saturation 38°C were observed in 27 (61%, 25 (57% and 15 (34% episodes respectively. Leucocytosis (WBC>11,000/ml and raised CRP (>10 mg/l were observed in 38 (86% and 43 (97% cases of pneumonia respectively. The area under the ROC curve for CRP was 0.827 (95% CI 0.720, 0.933. The diagnostic cut-off for CRP with an acceptable sensitivity (>0.8 was 25.60 mg/L (Youden index (J 0.515; sensitivity 0.848; specificity 0.667. A cut-off of 64.65 mg/L had the highest diagnostic accuracy (J 0.562; sensitivity 0.636; specificity 0.926.Patients with severe stroke frequently do not manifest key diagnostic features of pneumonia such as pyrexia, cough and purulent sputum early in their illness. The most common signs in this group are new-onset crackles, tachypnoea and hypoxia. Our results suggest that a CRP >25 mg/L should prompt investigations for pneumonia while values >65 mg/L have the highest diagnostic accuracy to justify consideration of this threshold as a diagnostic marker of post-stroke pneumonia.

  10. Investigation of Chlamydophila spp. in dairy cows with reproductive disorders

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    Niskanen Rauni

    2008-09-01

    Full Text Available Abstract Background Reports worldwide indicate high prevalence of Chlamydophila spp. infection in cattle. To assess the prevalence in Sweden, 525 cows in 70 dairy herds with reproductive disorders was investigated. Methods To detect antibodies two commercially available kits were used. Moreover, 107 specimens, including vaginal swabs, organ tissues and milk were analysed by Polymerase Chain Reaction (PCR. Results Two (0.4% cows were seropositive in the Pourquier Cp. abortus ELISA. The seroprevalence with the Chekit ELISA was 28% with no difference between cases and controls. Five specimens were positive in real-time PCR and further analysed by nested PCR. Cp. pecorum was confirmed by partial omp1 DNA sequencing of the nested PCR product of vaginal swabs from control cows. Conclusion The results suggest that Cp. abortus infection is absent or rare in Swedish cows whereas Cp. pecorum is probably more spread. They also suggest that Chlamydophila spp. are not related to reproduction disorders in Swedish cattle.

  11. Chlamydia pneumoniae CopD translocator protein plays a critical role in type III secretion (T3S and infection.

    Directory of Open Access Journals (Sweden)

    David C Bulir

    Full Text Available Pathogenic Gram-negative bacteria use type III secretion (T3S to inject effector proteins into the host cell to create appropriate conditions for infection and intracellular replication. Chlamydia spp. are believed to use T3S to infect their host cell, and the translocator proteins are an essential component of this system. Chlamydia pneumoniae contains genes encoding two sets of translocator proteins; CopB and CopD, and CopB2 and CopD2. In this study, we identified novel interactions between CopD and three type III secretion proteins; namely, CopN, CdsN, and CdsF. We identified a CopD putative chaperone binding motif, PxLxxP, within the N-terminal region (CopD amino acids 120-125, which was necessary for interaction with its putative chaperone LcrH_1. Using size exclusion chromatography, we showed that CopD and LcrH_1 formed higher order structures in solution with CopD and LcrH_1 binding in a ratio of 1∶1, which is unique for T3SS translocator proteins. Lastly, we showed that antibodies to CopD reduced C. pneumoniae infectivity by >95%. Collectively, this data suggests that CopD plays a critical role in pathogenesis and likely functions as a hydrophobic translocator of the type III secretion system in Chlamydia pneumoniae.

  12. Mycoplasma pneumonia

    Science.gov (United States)

    Walking pneumonia; Community-acquired pneumonia - mycoplasma; Community-acquired pneumonia - atypical ... Mycoplasma pneumonia usually affects people younger than 40. People who live or work in crowded areas such as schools ...

  13. First isolation and characterization of Chlamydophila abortus from abortion tissues of sheep in Sardinia, Italy.

    Science.gov (United States)

    Chisu, Valentina; Porcu, Rosaura; Tanda, Antonio; Masala, Giovanna

    2013-01-01

    Chlamydophila abortus (C. abortus) is the responsible agent for chlamydial abortion [commonly known as Enzootic Abortion of Ewes (EAE)] and, as such, it causes major financial losses to the sheep industry worldwide. Isolation of the pathogen is considered the 'gold standard' and most sensitive method of detection for diagnosing chlamydial infection. So far, there has been no isolation of C. abortus from ovines in Sardinia, Italy. This letter describes the results of a study conducted on a total of 89 aborted samples (40 foetuses and 49 placentae) collected in 2010 in Northern Sardinia, Italy. Three placentae resulted PCR-positive when analyzed using the putative outer membrane protein (pmp) specific primers, the test lead to the identification and first isolation in cell culture of C. abortus. This letter to the editor describes the first isolation of C. abortus from ovine placentae and increases the knowledge of one of the agents that causes ovine abortion in Sardinia and, more generally, in the Mediterranean basin.

  14. Chlamydophila psittaci and Chlamydophila pecorum infections in goats and sheep in Egypt.

    Science.gov (United States)

    Osman, K M; Ali, H A; ElJakee, J A; Galal, H M

    2011-12-01

    The aim of this study was to investigate the epidemiology of chlamydiosis in free-ranging asymptomatic and diarrhoeic sheep and goats in Egypt. Faecal swabs were examined for the presence of Chlamydiae by culture in Vero cells and chick embryos, and staining with Giménez, direct fluorescein-conjugated monoclonal antibodies, and immunoperoxidase. Specific chlamydial DNA was identified by amplification of the omp2 gene. The asymptomatic goats were 50% positive for the presence of the omp2 gene of the family Chlamydiaceae, and all isolates were Chlamydophila psittaci. The percentage of diseased goats in which Chlamydiaceae were identified was 16.2%, and all were positive for Cp. psittaci. Of the asymptomatic sheep, 6.7% were positive for the omp2 gene of the family Chlamydiaceae, and again all were positive for Cp. psittaci. In contrast, 42.9% of the samples that were collected from the diseased sheep were positive for Chlamydiaceae, of which 25.7% were Cp. psittaci and 4.8% Cp. pecorum.

  15. New real-time PCR tests for species-specific detection of Chlamydophila psittaci and Chlamydophila abortus from tissue samples.

    Science.gov (United States)

    Pantchev, Alexandra; Sting, Reinhard; Bauerfeind, Rolf; Tyczka, Judith; Sachse, Konrad

    2009-08-01

    Chlamydophila psittaci and Chlamydophila abortus are the causative agents of avian chlamydiosis (psittacosis) and ovine enzootic abortion, respectively. Both pathogens are known to possess zoonotic potential. Due to their close genetic relatedness, direct and rapid species identification is difficult. In the present study, new real-time PCR assays are reported for both species. The tests are based on highly specific probes targeting the ompA gene region and were conducted as duplex PCRs including an internal amplification control. The Cp. psittaci assay successfully passed a proficiency test at national level. Examination of field samples revealed Cp. psittaci as the dominating species in birds, but also Cp. abortus in a few psittacines. Real-time PCR assays for species-specific detection of Cp. psittaci and Cp. abortus are suited for routine diagnosis, which renders them important tools for the recognition of outbreaks of psittacosis and ovine enzootic abortion.

  16. Cellular localization of choline-utilization proteins in Streptococcus pneumoniae using novel fluorescent reporter systems

    NARCIS (Netherlands)

    Eberhardt, Alice; Wu, Ling J.; Errington, Jeff; Vollmer, Waldemar; Veening, Jan-Willem

    2009-01-01

    P>The molecular mechanisms underlying cell growth, cell division and pathogenesis in Streptococcus pneumoniae are still not fully understood. Single-cell methodologies are potentially of great value to investigate S. pneumoniae cell biology. Here, we report the construction of novel plasmids for sin

  17. Molecular biology of Chlamydia pneumoniae surface proteins and their role in immunopathogenicity

    DEFF Research Database (Denmark)

    Christiansen, Gunna; Boesen, Thomas; Hjernø, Karin

    1999-01-01

    BACKGROUND: The association of Chlamydia pneumoniae with the development of atherosclerosis is based on serology and on detection of C pneumoniae-specific DNA by polymerase chain reaction in the atheromas. METHODS AND RESULTS: Because the humoral immune response frequently recognizes epitopes pre...

  18. Structural Study of MPN387, an Essential Protein for Gliding Motility of a Human-Pathogenic Bacterium, Mycoplasma pneumoniae.

    Science.gov (United States)

    Kawakita, Yoshito; Kinoshita, Miki; Furukawa, Yukio; Tulum, Isil; Tahara, Yuhei O; Katayama, Eisaku; Namba, Keiichi; Miyata, Makoto

    2016-09-01

    Mycoplasma pneumoniae is a human pathogen that glides on host cell surfaces with repeated catch and release of sialylated oligosaccharides. At a pole, this organism forms a protrusion called the attachment organelle, which is composed of surface structures, including P1 adhesin and the internal core structure. The core structure can be divided into three parts, the terminal button, paired plates, and bowl complex, aligned in that order from the front end of the protrusion. To elucidate the gliding mechanism, we focused on MPN387, a component protein of the bowl complex which is essential for gliding but dispensable for cytadherence. The predicted amino acid sequence showed that the protein features a coiled-coil region spanning residue 72 to residue 290 of the total of 358 amino acids in the protein. Recombinant MPN387 proteins were isolated with and without an enhanced yellow fluorescent protein (EYFP) fusion tag and analyzed by gel filtration chromatography, circular dichroism spectroscopy, analytical ultracentrifugation, partial proteolysis, and rotary-shadowing electron microscopy. The results showed that MPN387 is a dumbbell-shaped homodimer that is about 42.7 nm in length and 9.1 nm in diameter and includes a 24.5-nm-long central parallel coiled-coil part. The molecular image was superimposed onto the electron micrograph based on the localizing position mapped by fluorescent protein tagging. A proposed role of this protein in the gliding mechanism is discussed. Human mycoplasma pneumonia is caused by a pathogenic bacterium, Mycoplasma pneumoniae This tiny, 2-μm-long bacterium is suggested to infect humans by gliding on the surface of the trachea through binding to sialylated oligosaccharides. The mechanism underlying mycoplasma "gliding motility" is not related to any other well-studied motility systems, such as bacterial flagella and eukaryotic motor proteins. Here, we isolated and analyzed the structure of a key protein which is directly involved in the

  19. A novel typing method for Streptococcus pneumoniae using selected surface proteins

    Directory of Open Access Journals (Sweden)

    Arnau eDomenech

    2016-03-01

    Full Text Available The diverse pneumococcal diseases are associated with different pneumococcal lineages, or clonal complexes. Nevertheless, intra-clonal genomic variability, which influences pathogenicity, has been reported for surface virulence factors. These factors constitute the communication interface between the pathogen and its host and their corresponding genes are subjected to strong selective pressures affecting functionality and immunogenicity. First, the presence and allelic dispersion of 97 outer protein families were screened in 19 complete pneumococcal genomes. Seventeen families were deemed variable and were then examined in 216 draft genomes. This procedure allowed the generation of binary vectors with 17 positions and the classification of strains into surfotypes. They represent the outer protein subsets with the highest inter-strain discriminative power. A total of 116 non-redundant surfotypes were identified. Those sharing a critical number of common protein features were hierarchically clustered into 18 surfogroups. Most clonal complexes with comparable epidemiological characteristics belonged to the same or similar surfogroups. However, the very large CC156 clonal complex was dispersed over several surfogroups. In order to establish a relationship between surfogroup and pathogenicity, the surfotypes of 95 clinical isolates with different serogroup/serotype combinations were analysed. We found a significant correlation between surfogroup and type of pathogenic behaviour (primary invasive, opportunistic invasive and non-invasive. We conclude that the virulent behaviour of S. pneumoniae is related to the activity of collections of, rather than individual, surface virulence factors. Since surfotypes evolve faster than MLSTs and directly reflect virulence potential, this novel typing protocol is appropriate for the identification of emerging clones.

  20. CPSIT_p7重组表达产物免疫原性检测及其在鹦鹉热嗜衣原体持续性感染中表达的研究%Immunogenicity analysis of a recombinant protein CPSIT_p7 and its expression in HeLa cells during persistent Chlamydophila psittaci infection

    Institute of Scientific and Technical Information of China (English)

    贺庆芝; 曾怀才; 黎知青; 王川; 胡艳群; 陈芝喜; 吴移谋

    2014-01-01

    目的:构建鹦鹉热嗜衣原体( Chlamydophila psittaci,Cps) CPSIT_p7原核表达载体,表达并纯化CPSIT_p7,检测其免疫原性,观察其在持续性感染过程中的mRNA和蛋白动态表达情况。方法原核表达和纯化His-CPSIT_p7融合蛋白,免疫BALB/c小鼠制备多克隆抗体,ELISA分析His-CPSIT_p7蛋白的免疫原性。采用青霉素与Cps共同处理细胞来制备持续性感染模型, RT-PCR和Western blot检测CPSIT_p7在其持续性感染过程中的表达情况。结果成功构建了pET30a-CPSIT_p7原核表达载体,并表达和纯化较稳定的重组蛋白;该蛋白免疫小鼠40 d后,ELISA 检测血清特异性抗体效价为1∶1000000;RT-PCR和Western blot 结果显示在Cps急性感染与持续性感染过程中CPSIT_p7 mRNA和蛋白表达水平均呈时间依赖性增加,感染后mRNA在36 h表达量达到高峰,之后急性感染呈时间依赖性下降,而持续性感染CPSIT_p7 mRNA高表达水平至少持续到60 h。结论成功克隆表达了His-CPSIT_p7,该蛋白具有较好的免疫原性;CPSIT_p7基因和蛋白在Cps持续性感染中呈高表达。%Objective To construct a prokaryotic expression plasmid for CPSIT_p7 gene from Chlamydophila psittaci ( Cps) 6BC strain and to evaluate immunogenicity of the recombinant protein His-CPSIT_p7 and detect its dynamic expression at mRNA and protein levels in HeLa cells during persistent Cps infection.Methods The fusion protein His-CPSIT_p7 was expressed in E.coli BL21 and purified by Ni-NTA affinity chromatography .BALB/c mice were immunized with the recombinant protein to prepare polyclonal antibody for evaluation of the immunogenicity of His-CPSIT_p7 by ELISA.Penicillin sodium was used to establish a model of Cps persistence infection .RT-PCR and Western blot assay were performed to de-tect the expression of CPSIT_p7 at mRNA and protein levels during Cps persistent infection .Results The fusion protein His-CPSIT_p7 was successfully

  1. Inhibition of Streptococcus pneumoniae penicillin-binding protein 2x and Actinomadura R39 DD-peptidase activities by ceftaroline.

    Science.gov (United States)

    Zervosen, Astrid; Zapun, André; Frère, Jean-Marie

    2013-01-01

    Although the rate of acylation of a penicillin-resistant form of Streptococcus pneumoniae penicillin-binding protein 2x (PBP2x) by ceftaroline is 80-fold lower than that of its penicillin-sensitive counterpart, it remains sufficiently high (k(2)/K = 12,600 M(-1) s(-1)) to explain the sensitivity of the penicillin-resistant strain to this new cephalosporin. Surprisingly, the Actinomadura R39 DD-peptidase is not very sensitive to ceftaroline.

  2. Differential Regulation of Protein- and Polysaccharide-Specific Ig Isotype Production In Vivo in Response to Intact Streptococcus pneumoniae

    Science.gov (United States)

    2006-01-01

    whether DCs played a role in either or both of these responses. We first demonstrated that immature bone marrow-derived myeloid dendritic cells ( BmDC ...1640 E-mail: csnapper@usuhs.mil Keywords: Streptococcus pneumoniae, immunoglobulin isotypes, murine, T cells , dendritic cells , cytokines, Toll...polysaccharide; PC, phosphorylcholine; PspA, pneumococcal surface protein A; DC, dendritic cell ; TLR, Toll-like receptor; TI, T cell -independent

  3. Affinity of ceftaroline and other beta-lactams for penicillin-binding proteins from Staphylococcus aureus and Streptococcus pneumoniae.

    Science.gov (United States)

    Kosowska-Shick, K; McGhee, P L; Appelbaum, P C

    2010-05-01

    We compared the affinities of ceftaroline for all penicillin-binding proteins (PBPs) with those of ceftriaxone and cefotaxime in 6 Staphylococcus aureus and 7 Streptococcus pneumoniae isolates with various resistance phenotypes. Ceftaroline MICs were pneumoniae. Ceftaroline affinities for penicillin-susceptible S. pneumoniae strains were in the order PBP2X and -3 > PBP1A, -1B, and -2A > PBP2B, and ceftaroline had >or=4-fold higher 50% inhibitory concentrations (IC(50)s) (0.1 to 4 microg/ml) for PBP2X, -2A, -2B, and -3 than those for the other cephalosporins tested. Among 3 penicillin-resistant S. pneumoniae strains, ceftaroline had a high affinity for PBP2X (IC(50), 0.1 to 1 microg/ml), a primary target for cephalosporin PBP binding activity, and high affinities for PBP2B (IC(50), 0.5 to 4 microg/ml) and PBP1A (IC(50), 0.125 to 0.25 microg/ml) as well, both of which are also known as major targets for PBP binding activity of cephalosporins. Ceftaroline PBP affinities in methicillin-susceptible S. aureus strains were greater than or equal to those of the 3 other beta-lactams tested. Ceftaroline bound to PBP2a in methicillin-resistant S. aureus (IC(50), 0.01 to 1 microg/ml) with up to 256-fold-higher affinity than those of other agents. Ceftaroline demonstrated very good PBP affinity against all S. aureus and S. pneumoniae strains tested, including resistant isolates.

  4. Antibodies to Chlamydia trachomatis heat shock proteins in women with tubal factor infertility are associated with prior infection by C. trachomatis but not by C. pneumoniae

    DEFF Research Database (Denmark)

    Persson, K; Osser, S; Birkelund, Svend

    1999-01-01

    proteins and to C. trachomatis but no independent influence of antibodies to C. pneumoniae. No interaction between C. trachomatis and C. pneumoniae suggesting a synergistic effect was found although the heat shock proteins from these two organisms are immunologically similar. Antibodies to the chlamydial......The antibody response to heat shock proteins 60 and 10 were studied in 163 patients with tubal factor infertility and in 163 age-matched pregnant women. The associations of these antibodies with specific antibodies to Chlamydia trachomatis and to Chlamydia pneumoniae as well as with antibodies...... to the common chlamydial lipopolysaccharide antigen were studied. Patients with tubal factor infertility had significantly higher frequencies and titres of all antibodies except to C. pneumoniae. In a logistic regression model an association was found between the prevalence of antibodies to the heat shock...

  5. An investigation into the role of Chlamydophila spp. in bovine upper respiratory tract disease.

    Science.gov (United States)

    Twomey, D F; Griffiths, P C; Horigan, M W; Hignett, B C; Martin, T P

    2006-05-01

    An outbreak of upper respiratory tract disease was investigated in a group of 17 housed home-bred calves on a mixed dairy, beef and sheep farm in Devon. Conjunctival swabs were collected and tested for Chlamydophila spp. DNA using a PCR test that detects Chlamydophila abortus and Chlamydophila psittaci. Six of the calves tested gave a positive result. Further epidemiological observations and laboratory testing indicated that the adult dairy cows, from which the affected calves originated, were the most likely source of infection.

  6. In vitro activities of demestic macrolides against Chlamydia trachomatis and Chlamydophila pneumoniae%四种国产大环内酯类抗菌药物体外抗沙眼衣原体和肺炎衣原体敏感性研究

    Institute of Scientific and Technical Information of China (English)

    倪安平; 崔京涛; 汪晓巍

    2008-01-01

    目的 研究国产必特螺旋霉素、乙酰螺旋霉素、红霉素和阿奇霉素等4种大环内酯类药物对于沙眼衣原体和肺炎衣原体体外药物敏感性试验,评估其抗衣原体作用,以指导临床用药.方法 细胞培养和免疫荧光包涵体染色技术测定4种国产大环内酯类抗菌药物对于沙眼衣原体和肺炎衣原体体外MIC.结果 对于沙眼血清型B,必特螺旋霉素、红霉素和阿奇霉素体外MIC为0.5μg/ml,乙酰螺旋霉素为4μg/ml.对于沙眼血清型D,必特螺旋霉素与阿奇霉素体外MIC均为0.25μg/ml,红霉素0.5μg/ml,乙酰螺旋霉素2μg/ml.对于肺炎衣原体,红霉素体外MIC≤0.016μg/ml,阿奇霉素和必特螺旋霉素均为0.032μg/ml,乙酰螺旋霉素0.5μg/ml.结论 国产必特螺旋霉素、红霉素和阿奇霉素体外抗沙眼衣原体(血清型B和D)和肺炎衣原体作用可靠,但乙酰螺旋霉素则较差.%Objective To evaluate the activities of four demestic macrolides against C. trachomatis and C. pneumoniae by antimicrobiai susceptibility testing. Methods Cell culture and immunoflourescence staining of chlamydial inclusions were used to determine MICs of four demestic macrolides against C. trachomatis and C. pneumoniae. Results MIC (0.5 μg/ml) was found for acylspriramycin,erythromycin and azithromycin against C. trachomatis serovar B while it was 4 μg/ml for acetylspiramycin. Agaisnt C. trachomatis serovar D, MIC was 0.25 μg/mi in both acylspriramycin and azithromycin, and MICs were 0.5 μg/ml and 2 μml in erythromycin and acetylspiramycin, separately. Agaisnt C. pneumoniae TWAR, erythromycin was the most active with MIC≤0. 016 μg/ml, acylspriramycin and azithromycin were the second with same M1C of 0.032 μg/ml. However, acetylspiramycin was less active with 0.5 μg/ml of MIC. Conclusion Except acetylspiramycin, acylspriramycin erythromycin and azithromycin had reliable activities against both C. trachomatis (serovar B and D) and C. pneumoniae.

  7. Chlamydophila psittaci infection of birds and humans

    Directory of Open Access Journals (Sweden)

    Bülent Baş

    2015-04-01

    seen in infected birds. Transmission of infection to humans occurs through inhalation or direct contact and transmission through bird bites or human-to-human is rare. C. psittaci usually leads to the systemic infection associated with pneumonia in humans. In recent years, PCR based molecular methods are used as well as serological methods such as ELISA, CFT, MIF in diagnosis. Both of infected birds and humans, tetracyclines and macrolides are preferred for treatment of infection. In order to prevent the disease, due to there isn't any commercial vaccine for especially using in birds, applying biosafety rules is very important in terms of human health and economical aspects. Especially, veterinarians, bird breeders and dealers, poultry farmers and slaughterhouse workers are at high risk for C. psittaci infection. Due to the transmission to humans of psittacosis infection and accepting it as a potential biological weapon, it is thought to be important for public health. In this review, it is aimed to give detailed information about infection in human and birds, because it can be missed at the diagnosis, hence there is low awareness about disease and it has got variable clinical symptoms.

  8. Correlation of Chlamydia and Chlamydophila spp. IgG and IgM antibodies by microimmunofluorescence with antigen detection methods.

    Science.gov (United States)

    She, Rosemary C; Welch, Ryan; Wilson, Andrew R; Davis, David; Litwin, Christine M

    2011-01-01

    Correlation of serologic titers for Chlamydia trachomatis with other tests has been based on direct fluorescence antibody (DFA) testing and culture, but not on nucleic acid-based tests that are used for screening. We retrospectively reviewed the specificity of antibodies against C. trachomatis, Chlamydia psittaci, and Chlamydophila pneumoniae by microimmunofluorescence (MIF) when compared with DFA, culture, nucleic acid probe, and transcription-mediated amplification. Over a 6-year period, 226 cases had both MIF and one of these other methods performed for comparison. Agreement between C. trachomatis antigen or nucleic acid detection and MIF results was 87% (197/226). C. trachomatis serology had a negative predictive value of 98%, and 10.6% of cases were positive by serology and negative by antigen testing. Of the 13 patients who had a positive C. trachomatis antigen or nucleic acid test result, 9 had IgG and/or IgM titers highest against C. trachomatis, 3 had IgG titers highest against C. pneumoniae, and 1 had undetectable titers for the three chlamydial species. Twenty-five patients had positive IgG and/or IgM titers to C. trachomatis but negative antigen test results. Serologic testing can increase the sensitivity of detecting C. trachomatis infections.

  9. Chlamydial development is blocked in host cells transfected with Chlamydophila caviae incA.

    Science.gov (United States)

    Alzhanov, Damir; Barnes, Jennifer; Hruby, Dennis E; Rockey, Daniel D

    2004-07-01

    Chlamydiae produce a set of proteins, termed Inc proteins, that are localized to the inclusion membrane and exposed to the host cell cytosol. Little information exists regarding the interaction of Inc proteins with the eukaryotic cell. To examine these interactions, Vaccinia virus vectors and mammalian plasmid-based systems were used to express inc genes in mammalian cells. Cells transfected with plasmids expressing Chlamydophila caviae incA were not productively infected by C. caviae. Expression of C. caviae incA also reduced inclusion formation by Chlamydia trachomatis, but not to the degree seen for C. caviae. Chlamydia trachomatis incA did not block development of either C. trachomatis or C. caviae. Deletion mutagenesis was used to demonstrate that plasmids encoding either the amino or carboxy-terminal regions of the protein, as well as the changing of a single amino acid within IncA (serine 17) could not block C. caviae infection. Immunoblot analysis of truncated IncA in a Vaccinia virus system provided evidence that serine 17 of C. caviae IncA is a target for phosphorylation. These experiments provide insight into the interaction of Inc proteins with the host cell and introduce a model system where these interactions can be explored further.

  10. Chlamydial development is blocked in host cells transfected with Chlamydophila caviae incA

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    Barnes Jennifer

    2004-07-01

    Full Text Available Abstract Background Chlamydiae produce a set of proteins, termed Inc proteins, that are localized to the inclusion membrane and exposed to the host cell cytosol. Little information exists regarding the interaction of Inc proteins with the eukaryotic cell. To examine these interactions, Vaccinia virus vectors and mammalian plasmid-based systems were used to express inc genes in mammalian cells. Results Cells transfected with plasmids expressing Chlamydophila caviae incA were not productively infected by C. caviae. Expression of C. caviae incA also reduced inclusion formation by Chlamydia trachomatis, but not to the degree seen for C. caviae. Chlamydia trachomatis incA did not block development of either C. trachomatis or C. caviae. Deletion mutagenesis was used to demonstrate that plasmids encoding either the amino or carboxy-terminal regions of the protein, as well as the changing of a single amino acid within IncA (serine 17 could not block C. caviae infection. Immunoblot analysis of truncated IncA in a Vaccinia virus system provided evidence that serine 17 of C. caviae IncA is a target for phosphorylation. Conclusions These experiments provide insight into the interaction of Inc proteins with the host cell and introduce a model system where these interactions can be explored further.

  11. Surfactant protein-A suppresses eosinophil-mediated killing of Mycoplasma pneumoniae in allergic lungs.

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    Julie G Ledford

    Full Text Available Surfactant protein-A (SP-A has well-established functions in reducing bacterial and viral infections but its role in chronic lung diseases such as asthma is unclear. Mycoplasma pneumoniae (Mp frequently colonizes the airways of chronic asthmatics and is thought to contribute to exacerbations of asthma. Our lab has previously reported that during Mp infection of non-allergic airways, SP-A aides in maintaining airway homeostasis by inhibiting an overzealous TNF-alpha mediated response and, in allergic mice, SP-A regulates eosinophilic infiltration and inflammation of the airway. In the current study, we used an in vivo model with wild type (WT and SP-A(-/- allergic mice challenged with the model antigen ovalbumin (Ova that were concurrently infected with Mp (Ova+Mp to test the hypothesis that SP-A ameliorates Mp-induced stimulation of eosinophils. Thus, SP-A could protect allergic airways from injury due to release of eosinophil inflammatory products. SP-A deficient mice exhibit significant increases in inflammatory cells, mucus production and lung damage during concurrent allergic airway disease and infection (Ova+Mp as compared to the WT mice of the same treatment group. In contrast, SP-A deficient mice have significantly decreased Mp burden compared to WT mice. The eosinophil specific factor, eosinophil peroxidase (EPO, which has been implicated in pathogen killing and also in epithelial dysfunction due to oxidative damage of resident lung proteins, is enhanced in samples from allergic/infected SP-A(-/- mice as compared to WT mice. In vitro experiments using purified eosinophils and human SP-A suggest that SP-A limits the release of EPO from Mp-stimulated eosinophils thereby reducing their killing capacity. These findings are the first to demonstrate that although SP-A interferes with eosinophil-mediated biologic clearance of Mp by mediating the interaction of Mp with eosinophils, SP-A simultaneously benefits the airway by limiting inflammation

  12. Klebsiella pneumoniae O antigen loss alters the outer membrane protein composition and the selective packaging of proteins into secreted outer membrane vesicles.

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    Cahill, Bethaney K; Seeley, Kent W; Gutel, Dedra; Ellis, Terri N

    2015-11-01

    Klebsiella pneumoniae is a nosocomial pathogen which naturally secretes lipopolysaccharide (LPS) and cell envelope associated proteins into the environment through the production of outer membrane vesicles (OMVs). The loss of the LPS O antigen has been demonstrated in other bacterial species to significantly alter the composition of OMVs. Therefore, this study aimed to comprehensively analyze the impact of O antigen loss on the sub-proteomes of both the outer membrane and secreted OMVs from K. pneumoniae. As determined by LC-MS/MS, OMVs were highly enriched with outer membrane proteins involved in cell wall, membrane, and envelope biogenesis as compared to the source cellular outer membrane. Deletion of wbbO, the enzyme responsible for O antigen attachment to LPS, decreased but did not eliminate this enrichment effect. Additionally, loss of O antigen resulted in OMVs with increased numbers of proteins involved in post-translational modification, protein turnover, and chaperones as compared to secreted vesicles from the wild type. This alteration of OMV composition may be a compensatory mechanism to deal with envelope stress. This comprehensive analysis confirms the highly distinct protein composition of OMVs as compared to their source membrane, and provides evidence for a selective sorting mechanism that involves LPS polysaccharides. These data support the hypothesis that modifications to LPS alters both the mechanics of protein sorting and the contents of secreted OMVs and significantly impacts the protein composition of the outer membrane.

  13. Vaccination with a Streptococcus pneumoniae trivalent recombinant PcpA, PhtD and PlyD1 protein vaccine candidate protects against lethal pneumonia in an infant murine model.

    Science.gov (United States)

    Verhoeven, David; Xu, Qingfu; Pichichero, Michael E

    2014-05-30

    Streptococcus pneumoniae infections continue to cause significant worldwide morbidity and mortality despite the availability of efficacious serotype-dependent vaccines. The need to incorporate emergent strains expressing additional serotypes into pneumococcal polysaccharide conjugate vaccines has led to an identified need for a pneumococcal protein-based vaccine effective against a broad scope of serotypes. A vaccine consisting of several conserved proteins with different functions during pathogenesis would be preferred. Here, we investigated the efficacy of a trivalent recombinant protein vaccine containing pneumococcal choline-binding protein A (PcpA), pneumococcal histidine triad D (PhtD), and genetically detoxified pneumolysin (PlyD1) in an infant mouse model. We found the trivalent vaccine conferred protection from lethal pneumonia challenges using serotypes 6A and 3. The observed protection with trivalent PcpA, PhtD, and PlyD1 vaccine in infant mice supports the ongoing study of this candidate vaccine in human infant clinical trials.

  14. Antibodies Directed against a Peptide Epitope of a Klebsiella pneumoniae-Derived Protein Are Present in Ankylosing Spondylitis

    Science.gov (United States)

    Tinazzi, Elisa; Moretta, Francesca; D’Angelo, Salvatore; Olivieri, Ignazio; Lunardi, Claudio

    2017-01-01

    Ankylosing spondylitis (AS) is a chronic inflammatory arthritis of unknown origin. Its autoimmune origin has been suggested but never proven. Several reports have implicated Klebsiella pneumoniae as a triggering or perpetuating factor in AS; however, its role in the disease pathogenesis remains debated. Moreover, despite extensive investigations, a biomarker for AS has not yet been identified. To clarify these issues, we screened a random peptide library with pooled IgGs obtained from 40 patients with AS. A peptide (AS peptide) selected from the library was recognized by serum IgGs from 170 of 200 (85%) patients with AS but not by serum specimens from 100 healthy controls. Interestingly, the AS peptide shows a sequence similarity with several molecules expressed at the fibrocartilaginous sites that are primarily involved in the AS inflammatory process. Moreover, the peptide is highly homologous to a Klebsiella pneumoniae dipeptidase (DPP) protein. The antibody affinity purified against the AS peptide recognizes the autoantigens and the DPP protein. Furthermore, serum IgG antibodies against the Klebsiella DPP121-145 peptide epitope were detected in 190 of 200 patients with AS (95%), 3 of 200 patients with rheumatoid arthritis (1.5%) and only 1 of 100 (1%) patients with psoriatic arthritis. Such reactivity was not detected in healthy control donors. Our results show that antibodies directed against an epitope of a Klebsiella pneumoniae-derived protein are present in nearly all patients with AS. In the absence of serological biomarkers for AS, such antibodies may represent a useful tool in the diagnosis of the disease. PMID:28135336

  15. C-reactive protein (CRP): an important diagnostic and prognostic tool in nursing-home-associated pneumonia.

    Science.gov (United States)

    Arinzon, Zeev; Peisakh, Alexander; Schrire, Samuel; Berner, Yitshal

    2011-01-01

    Pneumonia is the second most common infection in long term care (LTC) residents and is a leading cause of death from infection in those groups of patients. Atypical presentations and fewer presenting signs and symptoms in older patients complicate diagnosis and delay initiation of adequate treatment. The aim of this study was to compare laboratory CRP levels to pneumonia severity scores, in prediction of short-term death from pneumonia. Diagnosis of pneumonia was performed according to the criteria of McGeer for the identification of pneumonia at an LTC facility. The severities of pneumonia and mortality prediction were assessed by three indices: PSI (pneumonia severity index), Missouri study index and the nursing home associated pneumonia (NHAP) severity index. A strong positive correlation was found between CRP levels and PSI (r=0.445, pnursing home acquired pneumonia is not specific, it is suggested that CRP should be performed in every patient with a suspicion of pneumonia.

  16. Affinity of ceftobiprole for penicillin-binding protein 2b in Streptococcus pneumoniae strains with various susceptibilities to penicillin.

    Science.gov (United States)

    Davies, Todd A; He, Wenping; Bush, Karen; Flamm, Robert K

    2010-10-01

    Wild-type penicillin-binding protein (PBP) 2b from penicillin-susceptible Streptococcus pneumoniae had high affinity for ceftobiprole and penicillin (50% inhibitory concentrations [IC(50)s] of ≤0.15 μg/ml) but not ceftriaxone (IC(50) of >8 μg/ml). In clinical isolates, ceftobiprole and PBP 2b affinities were reduced 15- to 30-fold with a Thr-446-Ala substitution and further still with an additional Ala-619-Gly PBP 2b substitution. Ceftobiprole remained active (MICs of ≤1 μg/ml) against all strains tested and behaved more like penicillin than ceftriaxone with respect to PBP 2b binding.

  17. Prognostic value of immunohistochemical surfactant protein A expression in regenerative/hyperplastic alveolar epithelial cells in idiopathic interstitial pneumonias

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    Kajiki Akira

    2011-03-01

    Full Text Available Abstract Background It is difficult to predict survival in patients with idiopathic pulmonary fibrosis. Recently, several proteins, such as surfactant protein (SP and KL-6, have been reported to be useful biologic markers for prediction of prognosis for interstitial pneumonias. It is not clear whether there is any relationship between expression of these proteins in regenerative/hyperplastic alveolar epithelial cells and prognosis of idiopathic interstitial pneumonias (IIPs. Objectives This study aimed to elucidate the clinical significance of the expression of such lung secretory proteins as SP-A and KL-6 in lung tissues of patients with IIPs. Methods We retrospectively investigated the immunohistochemical expression of SP-A, KL-6, cytokeratin (CK, and epithelial membrane antigen (EMA in alveolar epithelial cells in lung tissues obtained from surgical lung biopsy in 43 patients with IIPs, and analyzed the correlation between expression of these markers and the prognosis of each IIP patient. CK and EMA were used as general markers for epithelial cells. Results In patients with usual interstitial pneumonia (UIP, the ratio of SP-A positive epithelial cells to all alveolar epithelial cells (SP-A positive ratio in the collapsed and mural fibrosis areas varied, ranging from cases where almost all alveolar epithelial cells expressed SP-A to cases where only a few did. On the other hand, in many patients with nonspecific interstitial pneumonia (NSIP, many of the alveolar epithelial cells in the diseased areas expressed SP-A. The SP-A positive ratio was significantly lower in patients who died from progression of UIP than in patients with UIP who remained stable or deteriorated but did not die. In NSIP patients, a similar tendency was noted between the SP-A positive ratio and prognosis. Conclusions The results suggest that the paucity of immunohistochemical SP-A expression in alveolar epithelial cells in diseased areas (i.e. regenerative

  18. Crystallization and preliminary crystallographic analysis of the transpeptidase domain of penicillin-binding protein 2B from Streptococcus pneumoniae

    Energy Technology Data Exchange (ETDEWEB)

    Yamada, Mototsugu, E-mail: mototsugu-yamada@meiji.co.jp; Watanabe, Takashi; Baba, Nobuyoshi; Miyara, Takako; Saito, Jun; Takeuchi, Yasuo [Pharmaceutical Research Center, Meiji Seika Kaisha Ltd, 760 Morooka-cho, Kohoku-ku, Yokohama 222-8567 (Japan)

    2008-04-01

    The selenomethionyl-substituted transpeptidase domain of penicillin-binding protein (PBP) 2B from S. pneumoniae was isolated from a limited proteolysis digest of the soluble form of recombinant PBP 2B and then crystallized. MAD data were collected to 2.4 Å resolution. Penicillin-binding protein (PBP) 2B from Streptococcus pneumoniae catalyzes the cross-linking of peptidoglycan precursors that occurs during bacterial cell-wall biosynthesis. A selenomethionyl (SeMet) substituted PBP 2B transpeptidase domain was isolated from a limited proteolysis digest of a soluble form of recombinant PBP 2B and then crystallized. The crystals belonged to space group P4{sub 3}2{sub 1}2, with unit-cell parameters a = b = 86.39, c = 143.27 Å. Diffraction data were collected to 2.4 Å resolution using the BL32B2 beamline at SPring-8. The asymmetric unit contains one protein molecule and 63.7% solvent.

  19. High-mobility group nucleosome-binding domain 2 protein inhibits the invasion of Klebsiella pneumoniae into mouse lungs in vivo.

    Science.gov (United States)

    Zheng, Shuang; Ren, Laibin; Li, Heng; Shen, Xiaofei; Yang, Xiaolong; Li, Na; Wang, Xinyuan; Guo, Xiaojuan; Wang, Xiaoying; Huang, Ning

    2015-07-01

    Since bacterial invasion into host cells is a critical step in the infection process and the predominance of multiple-antibiotic-resistant Klebsiella (K.) pneumoniae strains, using molecular agents to interfere with K. pneumoniae invasion is an attractive approach for the prevention of infection and suppress the immune inflammatory response. In previous studies by our group, high-mobility group nucleosome-binding domain 2 (HMGN2) protein was shown to exhibit anti-bacterial activity in vitro. The objective of the present study was to investigate the effects of HMGN2 protein on the invasion of K. pneumoniae 03183 in vivo. The results showed that pre-treatment with 128 µg/ml HMGN2 significantly reduced K. pneumoniae 03183 invasion into mouse lungs and increased the mRNA expression of CXCL1 and LCN2 within 2 h. Immunohistochemical staining showed that F-actin expression was significantly decreased, and fluorescence microscopy and western blot analysis further demonstrated that HMGN2 significantly blocked K. pneumoniae 03183-induced actin polymerization. These changes implied that HMGN2 may provide protection against K. pneumoniae 03183 infection in vivo.

  20. Detection of antibody responses against Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis proteins in children with community-acquired pneumonia: effects of combining pneumococcal antigens, pre-existing antibody levels, sampling interval, age, and duration of illness.

    Science.gov (United States)

    Borges, I C; Andrade, D C; Vilas-Boas, A-L; Fontoura, M-S H; Laitinen, H; Ekström, N; Adrian, P V; Meinke, A; Cardoso, M-R A; Barral, A; Ruuskanen, O; Käyhty, H; Nascimento-Carvalho, C M

    2015-08-01

    We evaluated the effects of combining different numbers of pneumococcal antigens, pre-existing antibody levels, sampling interval, age, and duration of illness on the detection of IgG responses against eight Streptococcus pneumoniae proteins, three Haemophilus influenzae proteins, and five Moraxella catarrhalis proteins in 690 children aged pneumonia. Serological tests were performed on acute and convalescent serum samples with a multiplexed bead-based immunoassay. The median sampling interval was 19 days, the median age was 26.7 months, and the median duration of illness was 5 days. The rate of antibody responses was 15.4 % for at least one pneumococcal antigen, 5.8 % for H. influenzae, and 2.3 % for M. catarrhalis. The rate of antibody responses against each pneumococcal antigen varied from 3.5 to 7.1 %. By multivariate analysis, pre-existing antibody levels showed a negative association with the detection of antibody responses against pneumococcal and H. influenzae antigens; the sampling interval was positively associated with the detection of antibody responses against pneumococcal and H. influenzae antigens. A sampling interval of 3 weeks was the optimal cut-off for the detection of antibody responses against pneumococcal and H. influenzae proteins. Duration of illness was negatively associated with antibody responses against PspA. Age did not influence antibody responses against the investigated antigens. In conclusion, serological assays using combinations of different pneumococcal proteins detect a higher rate of antibody responses against S. pneumoniae compared to assays using a single pneumococcal protein. Pre-existing antibody levels and sampling interval influence the detection of antibody responses against pneumococcal and H. influenzae proteins. These factors should be considered when determining pneumonia etiology by serological methods in children.

  1. Subunits of the Pyruvate Dehydrogenase Cluster of Mycoplasma pneumoniae Are Surface-Displayed Proteins that Bind and Activate Human Plasminogen.

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    Anne Gründel

    Full Text Available The dual role of glycolytic enzymes in cytosol-located metabolic processes and in cell surface-mediated functions with an influence on virulence is described for various micro-organisms. Cell wall-less bacteria of the class Mollicutes including the common human pathogen Mycoplasma pneumoniae possess a reduced genome limiting the repertoire of virulence factors and metabolic pathways. After the initial contact of bacteria with cells of the respiratory epithelium via a specialized complex of adhesins and release of cell-damaging factors, surface-displayed glycolytic enzymes may facilitate the further interaction between host and microbe. In this study, we described detection of the four subunits of pyruvate dehydrogenase complex (PDHA-D among the cytosolic and membrane-associated proteins of M. pneumoniae. Subunits of PDH were cloned, expressed and purified to produce specific polyclonal guinea pig antisera. Using colony blotting, fractionation of total proteins and immunofluorescence experiments, the surface localization of PDHA-C was demonstrated. All recombinant PDH subunits are able to bind to HeLa cells and human plasminogen. These interactions can be specifically blocked by the corresponding polyclonal antisera. In addition, an influence of ionic interactions on PDHC-binding to plasminogen as well as of lysine residues on the association of PDHA-D with plasminogen was confirmed. The PDHB subunit was shown to activate plasminogen and the PDHB-plasminogen complex induces degradation of human fibrinogen. Hence, our data indicate that the surface-associated PDH subunits might play a role in the pathogenesis of M. pneumoniae infections by interaction with human plasminogen.

  2. Genotyping of Chlamydophila abortus strains by multilocus VNTR analysis.

    Science.gov (United States)

    Laroucau, Karine; Vorimore, Fabien; Bertin, Claire; Mohamad, Khalil Yousef; Thierry, Simon; Hermann, Willems; Maingourd, Cyril; Pourcel, Christine; Longbottom, David; Magnino, Simone; Sachse, Konrad; Vretou, Evangelia; Rodolakis, Annie

    2009-06-12

    Chlamydophila (C.) abortus is the causative agent of ovine enzootic abortion with zoonotic potential whose epidemiology has been held back because of the obligate intracellular habitat of the bacterium. In the present study, we report on a molecular typing method termed multiple loci variable number of tandem repeats (VNTR) Analysis (MLVA) for exploring the diversity of C. abortus. An initial analysis performed with 34 selected genetic loci on 34 ruminant strains including the variant Greek strains LLG and POS resulted in the identification of five polymorphic loci, confirming the widely held notion that C. abortus is a very homogeneous species. Analysis of additional 111 samples with the selected five loci resulted in the classification of all strains into six genotypes with distinct molecular patterns termed genotypes [1] through [6]. Interestingly, the classification of the isolates in the six genotypes was partly related to their geographical origin. Direct examination of clinical samples proved the MLVA to be suitable for direct typing. Analysis of the genomic sequences in six C. abortus prototypes of amplicons generated with each of the five selected VNTR primers revealed that variation between genotypes was caused by the presence or absence of coding tandem repeats in three loci. Amplification of Chlamydophila psittaci reference strains with the five selected VNTR primers and of the six C. abortus prototype strains with the eight VNTR primers established for the typing of C. psittaci [Laroucau, K., Thierry, S., Vorimore, F., Blanco, K., Kaleta, E., Hoop, R., Magnino, S., Vanrompay, D., Sachse, K., Myers, G.S., Bavoil, P.M., Vergnaud, G., Pourcel, C., 2008. High resolution typing of Chlamydophila psittaci by multilocus VNTR analysis (MLVA). Infect. Genet. Evol. 8(2), 171-181] showed that both MLVA typing systems were species-specific when all respective VNTR primer sets were used. In conclusion, the newly developed MLVA system provides a highly sensitive

  3. Crystallization and preliminary X-ray analysis of enoyl-acyl carrier protein reductase (FabK) from Streptococcus pneumoniae

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    Saito, Jun, E-mail: jun-saito@meiji.co.jp; Yamada, Mototsugu; Watanabe, Takashi; Kitagawa, Hideo; Takeuchi, Yasuo [Pharmaceutical Research Center, Meiji Seika Kaisha Ltd, 760 Morooka-cho, Kohoku-ku, Yokohama 222-8567 (Japan)

    2006-06-01

    Enoyl-acyl carrier protein (ACP) reductases are responsible for bacterial type II fatty-acid biosynthesis and are attractive targets for developing novel antibiotics. The S. pneumoniae enoyl-ACP reductase (FabK) was crystallized and selenomethionine MAD data were collected to 2 Å resolution. The enoyl-acyl carrier protein (ACP) reductase from Streptococcus pneumoniae (FabK; EC 1.3.1.9) is responsible for catalyzing the final step in each elongation cycle of fatty-acid biosynthesis. Selenomethionine-substituted FabK was purified and crystallized by the hanging-drop vapour-diffusion method at 277 K. The crystal belongs to space group P2{sub 1}, with unit-cell parameters a = 50.26, b = 126.70, c = 53.63 Å, β = 112.46°. Diffraction data were collected to 2.00 Å resolution using synchrotron beamline BL32B2 at SPring-8. Two molecules were estimated to be present in the asymmetric unit, with a solvent content of 45.1%.

  4. LocZ Is a New Cell Division Protein Involved in Proper Septum Placement in Streptococcus pneumoniae

    Science.gov (United States)

    Holečková, Nela; Molle, Virginie; Buriánková, Karolína; Benada, Oldřich; Kofroňová, Olga; Ulrych, Aleš; Branny, Pavel

    2014-01-01

    ABSTRACT How bacteria control proper septum placement at midcell, to guarantee the generation of identical daughter cells, is still largely unknown. Although different systems involved in the selection of the division site have been described in selected species, these do not appear to be widely conserved. Here, we report that LocZ (Spr0334), a newly identified cell division protein, is involved in proper septum placement in Streptococcus pneumoniae. We show that locZ is not essential but that its deletion results in cell division defects and shape deformation, causing cells to divide asymmetrically and generate unequally sized, occasionally anucleated, daughter cells. LocZ has a unique localization profile. It arrives early at midcell, before FtsZ and FtsA, and leaves the septum early, apparently moving along with the equatorial rings that mark the future division sites. Consistently, cells lacking LocZ also show misplacement of the Z-ring, suggesting that it could act as a positive regulator to determine septum placement. LocZ was identified as a substrate of the Ser/Thr protein kinase StkP, which regulates cell division in S. pneumoniae. Interestingly, homologues of LocZ are found only in streptococci, lactococci, and enterococci, indicating that this close phylogenetically related group of bacteria evolved a specific solution to spatially regulate cell division. PMID:25550321

  5. Plasma Monocyte Chemoattractant Protein-1 Level as a Predictor of the Severity of Community-Acquired Pneumonia.

    Science.gov (United States)

    Yong, Kok-Khun; Chang, Jer-Hwa; Chien, Ming-Hsien; Tsao, Shih-Ming; Yu, Ming-Chih; Bai, Kuan-Jen; Tsao, Thomas Chang-Yao; Yang, Shun-Fa

    2016-01-29

    Monocyte chemoattractant protein (MCP)-1 increases in the serum of immunocompetent patients with community-acquired pneumonia (CAP). However, the correlation between the circulating level of MCP-1 and severity of CAP remains unclear. This study investigated differential changes in the plasma MCP-1 levels of patients with CAP before and after an antibiotic treatment and further analyzes the association between the CAP severity and MCP-1 levels. We measured the plasma MCP-1 levels of 137 patients with CAP and 74 healthy controls by using a commercial enzyme-linked immunosorbent assay. Upon initial hospitalization, Acute Physiology and Chronic Health Evaluation II (APACHE II); confusion, urea level, respiratory rate, blood pressure, and age of >64 years (CURB-65); and pneumonia severity index (PSI) scores were determined for assessing the CAP severity in these patients. The antibiotic treatment reduced the number of white blood cells (WBCs) and neutrophils as well as the level of C-reactive protein (CRP) and MCP-1. The plasma MCP-1 level, but not the CRP level or WBC count, correlated with the CAP severity according to the PSI (r = 0.509, p < 0.001), CURB-65 (r = 0.468, p < 0.001), and APACHE II (r = 0.360, p < 0.001) scores. We concluded that MCP-1 levels act in the development of CAP and are involved in the severity of CAP.

  6. Immunization with Pneumococcal Surface Protein K of Nonencapsulated Streptococcus pneumoniae Provides Protection in a Mouse Model of Colonization.

    Science.gov (United States)

    Keller, Lance E; Luo, Xiao; Thornton, Justin A; Seo, Keun-Seok; Moon, Bo Youn; Robinson, D Ashley; McDaniel, Larry S

    2015-11-01

    Current vaccinations are effective against encapsulated strains of Streptococcus pneumoniae, but they do not protect against nonencapsulated Streptococcus pneumoniae (NESp), which is increasing in colonization and incidence of pneumococcal disease. Vaccination with pneumococcal proteins has been assessed for its ability to protect against pneumococcal disease, but several of these proteins are not expressed by NESp. Pneumococcal surface protein K (PspK), an NESp virulence factor, has not been assessed for immunogenic potential or host modulatory effects. Mammalian cytokine expression was determined in an in vivo mouse model and in an in vitro cell culture system. Systemic and mucosal mouse immunization studies were performed to determine the immunogenic potential of PspK. Murine serum and saliva were collected to quantitate specific antibody isotype responses and the ability of antibody and various proteins to inhibit epithelial cell adhesion. Host cytokine response was not reduced by PspK. NESp was able to colonize the mouse nasopharynx as effectively as encapsulated pneumococci. Systemic and mucosal immunization provided protection from colonization by PspK-positive (PspK(+)) NESp. Anti-PspK antibodies were recovered from immunized mice and significantly reduced the ability of NESp to adhere to human epithelial cells. A protein-based pneumococcal vaccine is needed to provide broad protection against encapsulated and nonencapsulated pneumococci in an era of increasing antibiotic resistance and vaccine escape mutants. We demonstrate that PspK may serve as an NESp target for next-generation pneumococcal vaccines. Immunization with PspK protected against pneumococcal colonization, which is requisite for pneumococcal disease.

  7. The Spl Serine Proteases Modulate Staphylococcus aureus Protein Production and Virulence in a Rabbit Model of Pneumonia

    Science.gov (United States)

    Salgado-Pabon, Wilmara; Meyerholz, David K.; White, Mark J.; Schlievert, Patrick M.

    2016-01-01

    ABSTRACT The Spl proteases are a group of six serine proteases that are encoded on the νSaβ pathogenicity island and are unique to Staphylococcus aureus. Despite their interesting biochemistry, their biological substrates and functions in virulence have been difficult to elucidate. We found that an spl operon mutant of the community-associated methicillin-resistant S. aureus USA300 strain LAC induced localized lung damage in a rabbit model of pneumonia, characterized by bronchopneumonia observed histologically. Disease in the mutant-infected rabbits was restricted in distribution compared to that in wild-type USA300-infected rabbits. We also found that SplA is able to cleave the mucin 16 glycoprotein from the surface of the CalU-3 lung cell line, suggesting a possible mechanism for wild-type USA300 spreading pneumonia to both lungs. Investigation of the secreted and surface proteomes of wild-type USA300 and the spl mutant revealed multiple alterations in metabolic proteins and virulence factors. This study demonstrates that the Spls modulate S. aureus physiology and virulence, identifies a human target of SplA, and suggests potential S. aureus targets of the Spl proteases. IMPORTANCE Staphylococcus aureus is a versatile human pathogen that produces an array of virulence factors, including several proteases. Of these, six proteases called the Spls are the least characterized. Previous evidence suggests that the Spls are expressed during human infection; however, their function is unknown. Our study shows that the Spls are required for S. aureus to cause disseminated lung damage during pneumonia. Further, we present the first example of a human protein cut by an Spl protease. Although the Spls were predicted not to cut staphylococcal proteins, we also show that an spl mutant has altered abundance of both secreted and surface-associated proteins. This work provides novel insight into the function of Spls during infection and their potential ability to degrade

  8. H-NS Nucleoid Protein Controls Virulence Features of Klebsiella pneumoniae by Regulating the Expression of Type 3 Pili and the Capsule Polysaccharide

    Science.gov (United States)

    Ares, Miguel A.; Fernández-Vázquez, José L.; Rosales-Reyes, Roberto; Jarillo-Quijada, Ma. Dolores; von Bargen, Kristine; Torres, Javier; González-y-Merchand, Jorge A.; Alcántar-Curiel, María D.; De la Cruz, Miguel A.

    2016-01-01

    Klebsiella pneumoniae is an opportunistic pathogen causing nosocomial infections. Main virulence determinants of K. pneumoniae are pili, capsular polysaccharide, lipopolysaccharide, and siderophores. The histone-like nucleoid-structuring protein (H-NS) is a pleiotropic regulator found in several gram-negative pathogens. It has functions both as an architectural component of the nucleoid and as a global regulator of gene expression. We generated a Δhns mutant and evaluated the role of the H-NS nucleoid protein on the virulence features of K. pneumoniae. A Δhns mutant down-regulated the mrkA pilin gene and biofilm formation was affected. In contrast, capsule expression was derepressed in the absence of H-NS conferring a hypermucoviscous phenotype. Moreover, H-NS deficiency affected the K. pneumoniae adherence to epithelial cells such as A549 and HeLa cells. In infection experiments using RAW264.7 and THP-1 differentiated macrophages, the Δhns mutant was less phagocytized than the wild-type strain. This phenotype was likely due to the low adherence to these phagocytic cells. Taken together, our data indicate that H-NS nucleoid protein is a crucial regulator of both T3P and CPS of K. pneumoniae. PMID:26904512

  9. 猪流产嗜性衣原体MOMP基因的克隆及原核表达%Cloning and prokaryotic expression of the major outer membrane protein coding gene of Swine Chlamydophila abortus

    Institute of Scientific and Technical Information of China (English)

    李西玉; 许信刚; 周继章; 邱昌庆; 曹小安; 宫晓炜; 王光华

    2010-01-01

    [目的]克隆猪流产嗜性衣原体青海株主要外膜蛋白(Major outer membrane protein,MOMP)基因,并进行序列分析及原核表达.[方法]根据GenBank公布的猪流产嗜性衣原体MOMP基因的核苷酸序列,设计并合成4条特异性引物,用套式PCR方法扩增猪流产嗜性衣原体青海株MOMP基因,将其克隆入pMD18-T载体中,进行测序及序列分析.然后将MOMP基因亚克隆入原核表达载体pGEX4T-1中,在大肠杆菌BL21(DE3)中用IPTG诱导表达,对表达产物进行SDS-PAGE和Western blot检测.[结果]扩增到猪流产嗜性衣原体青海株1170 bp的MOMP全长基因.序列分析结果表明,该基因与已发表的猪流产嗜性衣原体B11001株和CP/12株核苷酸同源性均为99.7%.SDS-PAGE电泳可检测到分子质量约为66 ku的融合蛋白,主要以包涵体形式存在.Western-blot分析表明,重组蛋白可被猪流产嗜性衣原体抗体识别.[结论]成功克隆了猪流产嗜性衣原体青海株MOMP基因,并进行了原核表达,表达的蛋白具有抗原活性.

  10. Molecular characteristics of penicillin-binding protein 2b, 2x and 1a sequences in Streptococcus pneumoniae isolates causing invasive diseases among children in Northeast China.

    Science.gov (United States)

    Zhou, X; Liu, J; Zhang, Z; Liu, Y; Wang, Y; Liu, Y

    2016-04-01

    Streptococcus pneumoniae is one of the common pathogens causing severe invasive infections in children. This study aimed to investigate the serotype distribution and variations of penicillin-binding proteins (PBPs) 2b, 2x and 1a in S. pneumoniae isolates causing invasive diseases in Northeast China. A total of 256 strains were isolated from children with invasive pneumococcal disease (IPD) from January 2000 to October 2014. All strains were serotyped and determined for antibiotic resistance. The amplicons of penicillin-binding domains in pbp1a, pbp2b and pbp2x genes were sequenced for variation identification. The most prevalent serotypes of isolates in IPD children were 19A, 14, 19F, 23F and 6B. 19A and 19F were the most frequent serotypes of penicillin-resistant S. pneumoniae (PRSP), which present with high resistance to amoxicillin, cefotaxime, ceftriaxone and meropenem. The numbers of amino acid substitutions of penicillin-non-susceptible S. pneumoniae (PNSP) isolates were higher than those of penicillin-sensitive S. pneumoniae isolates in all the PBP genes (p pneumoniae were closely associated with resistance to β-lactam antibiotics. This study provides new data for further monitoring of genetic changes related to the emergence and spread of resistance to β-lactam antibiotics in China.

  11. 肺炎衣原体和巨细胞病毒感染与颈动脉粥样硬化斑块及其稳定性的相关性%Correlations between chlamydophila pneumoniae, cytomegalovirus infection and carotid atherosclerotic plaque and its stability

    Institute of Scientific and Technical Information of China (English)

    曹磊光; 王涛

    2014-01-01

    目的 探讨颈动脉粥样硬化的发生与肺炎衣原体(Chlamydia pneumonia,Cpn)和巨细胞病毒(cytomegalovirus,CMV)两种微生物感染的相关性.方法 纳入颈动脉彩超显示内膜-中膜厚度(intima-media thickness,IMT)>1.5 mm的患者作为颈动脉粥样硬化组,而IMT< 1.0 mm的健康体检者作为对照组.采用酶联免疫吸附法检测血清抗Cpn和抗CMV IgG抗体水平和阳性率.比较颈动脉粥样硬化组与对照组的人口统计学、血管危险因素以及抗Cpn和抗CMV IgG抗体阳性率.结果 颈动脉粥样硬化组共纳入患者92例,其中稳定斑块患者30例,不稳定斑块患者62例;对照组共纳入49例健康体检者.颈动脉粥样硬化组年龄以及男性、高血压、糖尿病、高脂血症和吸烟患者的构成比与对照组均无显著性差异(P均>0.05).颈动脉粥样硬化组抗Cpn IgG(69.5%对26.5%彩=23.887,P<0.001)、抗CMV IgG(75.0%对30.6%;x2 =26.156,P<0.001)阳性率以及抗Cpn IgG+抗CMV IgG阳性率(51.2%对10.2%;x2 =24.006,P<0.001)均显著性高于对照组.在颈动脉粥样硬化组中,不稳定斑块亚组年龄以及男性、高血压、糖尿病、高脂血症和吸烟患者的构成比与稳定斑块亚组均无显著性差异(P均>0.05).不稳定斑块亚组抗Cpn IgG(80.6%对46.7%;x2=11.025,P=0.001)和抗CMV IgG(83.9%对56.7%;x2=7.980,P=0.005)阳性率以及抗Cpn IgG+抗CMV IgG阳性率(44.6%对7.6%;x2=10.210,P=0.006)显著性高于稳定斑块亚组.结论 颈动脉粥样硬化的发生和斑块稳定性均与Cpn和CMV感染相关,Cpn和CMV感染可能是颈动脉粥样硬化发生和斑块不稳定的重要因素.%Objective To investigate the correlations between the occurrence of carotid artery atherosclerosis and chlamydia pneumoniae (Cpn) and cytomegalovirus (CMV).Methods Carotid color ultrasonography showed that the patients with intima-media thickness (IMT) > 1.5 mm were enrolled and were used as a carotid atherosclerosis

  12. Another turn of the screw in shaving Gram-positive bacteria: Optimization of proteomics surface protein identification in Streptococcus pneumoniae.

    Science.gov (United States)

    Olaya-Abril, Alfonso; Gómez-Gascón, Lidia; Jiménez-Munguía, Irene; Obando, Ignacio; Rodríguez-Ortega, Manuel J

    2012-06-27

    Bacterial surface proteins are of outmost importance as they play critical roles in the interaction between cells and their environment. In addition, they can be targets of either vaccines or antibodies. Proteomic analysis through "shaving" live cells with proteases has become a successful approach for a fast and reliable identification of surface proteins. However, this protocol has not been able to reach the goal of excluding cytoplasmic contamination, as cell lysis is an inherent process during culture and experimental manipulation. In this work, we carried out the optimization of the "shaving" strategy for the Gram-positive human pathogen Streptococcus pneumoniae, a bacterium highly susceptible to autolysis, and set up the conditions for maximizing the identification of surface proteins containing sorting or exporting signals, and for minimizing cytoplasmic contamination. We also demonstrate that cell lysis is an inherent process during culture and experimental manipulation, and that a low level of lysis is enough to contaminate a "surfome" preparation with peptides derived from cytoplasmic proteins. When the optimized conditions were applied to several clinical isolates, we found the majority of the proteins described to induce protection against pneumococcal infection. In addition, we found other proteins whose protection capacity has not been yet tested. In addition, we show the utility of this approach for providing antigens that can be used in serological tests for the diagnosis of pneumococcal disease.

  13. Members of the Pmp protein family of Chlamydia pneumoniae mediate adhesion to human cells via short repetitive peptide motifs.

    Science.gov (United States)

    Mölleken, Katja; Schmidt, Eleni; Hegemann, Johannes H

    2010-11-01

    Chlamydiae sp. are obligate intracellular pathogens that cause a variety of diseases in humans. Adhesion of the infectious elementary body to the eukaryotic host cell is a pivotal step in chlamydial pathogenesis. Here we describe the characterization of members of the polymorphic membrane protein family (Pmp), the largest protein family (with up to 21 members) unique to Chlamydiaceae. We show that yeast cells displaying Pmp6, Pmp20 or Pmp21 on their surfaces, or beads coated with the recombinant proteins, adhere to human epithelial cells. A hallmark of the Pmp protein family is the presence of multiple repeats of the tetrapeptide motifs FxxN and GGA(I, L, V) and deletion analysis shows that at least two copies of these motifs are needed for adhesion. Importantly, pre-treatment of human cells with recombinant Pmp6, Pmp20 or Pmp21 protein reduces infectivity upon subsequent challenge with Chlamydia pneumoniae and correlates with diminished attachment of Chlamydiae to target cells. Antibodies specific for Pmp21 can neutralize infection in vitro. Finally, a combination of two different Pmp proteins in infection blockage experiments shows additive effects, possibly suggesting similar functions. Our findings imply that Pmp6, Pmp20 and Pmp21 act as adhesins, are vital during infection and thus represent promising vaccine candidates.

  14. Clinical Usefulness of Urinary Fatty Acid Binding Proteins in Assessing the Severity and Predicting Treatment Response of Pneumonia in Critically Ill Patients: A Cross-Sectional Study.

    Science.gov (United States)

    Tsao, Tsung-Cheng; Tsai, Han-Chen; Chang, Shi-Chuan

    2016-05-01

    To investigate the clinical relevance of urinary fatty acid binding proteins (FABPs), including intestinal-FABP, adipocyte-FABP, liver-FABP, and heart-FABP in pneumonia patients required admission to respiratory intensive care unit (RICU).Consecutive pneumonia patients who admitted to RICU from September 2013 to October 2014 were enrolled except for those with pneumonia for more than 24 h before admission to RICU. Pneumonia patients were further divided into with and without septic shock subgroups. Twelve patients without infection were enrolled to serve as control group. Urine samples were collected on days 1 and 7 after admission to RICU for measuring FABPs and inflammatory cytokines. Clinical and laboratory data were collected and compared between pneumonia and control groups, and between the pneumonia patients with and without septic shock.There were no significant differences in urinary levels of various FABPs and inflammatory cytokines measured on day 1 between control and pneumonia groups. Urinary values of intestine-FABP (P = 0.020), adipocyte-FABP (P = 0.005), heart-FABP (P = 0.025), and interleukin-6 (P = 0.019) were significantly higher and arterial oxygen tension/fraction of inspired oxygen (PaO2/FiO2, P/F) ratio (P = 0.024) was significantly lower in pneumonia patients with septic shock on day 1 than in those without septic shock. After multivariate analysis, adipocyte-FABP was the independent factor (P = 0.026). Urinary levels of FABPs measured on day 7 of pneumonia patients were significantly lower in the improved than in nonimproved groups (P = 0.030 for intestine-FABP, P = 0.003 for adipocyte-FABP, P = 0.010 for heart-FABP, and P = 0.008 for liver-FABP, respectively). After multivariate analysis, adipocyte-FABP was the independent factor (P = 0.023).For pneumonia patients required admission to RICU, urinary levels of adipocyte-FABP on days 1 and 7 after admission to RICU may be valuable in assessing the

  15. Overexpression of the endothelial protein C receptor is detrimental during pneumonia-derived gram-negative sepsis (Melioidosis.

    Directory of Open Access Journals (Sweden)

    Liesbeth M Kager

    Full Text Available The endothelial protein C receptor (EPCR enhances anticoagulation by accelerating activation of protein C to activated protein C (APC and mediates anti-inflammatory effects by facilitating APC-mediated signaling via protease activated receptor-1. We studied the role of EPCR in the host response during pneumonia-derived sepsis instigated by Burkholderia (B. pseudomallei, the causative agent of melioidosis, a common form of community-acquired Gram-negative (pneumosepsis in South-East Asia.Soluble EPCR was measured in plasma of patients with septic culture-proven melioidosis and healthy controls. Experimental melioidosis was induced by intranasal inoculation of B. pseudomallei in wild-type (WT mice and mice with either EPCR-overexpression (Tie2-EPCR or EPCR-deficiency (EPCR(-/-. Mice were sacrificed after 24, 48 or 72 hours. Organs and plasma were harvested to measure colony forming units, cellular influxes, cytokine levels and coagulation parameters. Plasma EPCR-levels were higher in melioidosis patients than in healthy controls and associated with an increased mortality. Tie2-EPCR mice demonstrated enhanced bacterial growth and dissemination to distant organs during experimental melioidosis, accompanied by increased lung damage, neutrophil influx and cytokine production, and attenuated coagulation activation. EPCR(-/- mice had an unremarkable response to B. pseudomallei infection as compared to WT mice, except for a difference in coagulation activation in plasma.Increased EPCR-levels correlate with accelerated mortality in patients with melioidosis. In mice, transgenic overexpression of EPCR aggravates outcome during Gram-negative pneumonia-derived sepsis caused by B. pseudomallei, while endogenous EPCR does not impact on the host response. These results add to a better understanding of the regulation of coagulation during severe (pneumosepsis.

  16. Intragenic complementation by the nifJ-coded protein of Klebsiella pneumoniae.

    OpenAIRE

    Stacey, G.; Zhu, J.; Shah, V. K.; Shen, S C; Brill, W J

    1982-01-01

    A single mutation, nifC1005 (Jin et al. Sci. Sin. 23:108-118, 1980), located between nifH and nifJ in the nif cluster of Klebsiella pneumoniae, genetically complemented mutations in each of the 17 known nif genes. This suggested that the mutation is located in a new nif gene. We showed by complementation analyses that only 3 of 12 nifJ mutations tested were complemented by nifC1005. Nitrogenase activity in cell extracts of the mutant with nifC1005 as well as NifJ- mutants was stimulated by th...

  17. Intragenic complementation by the nifJ-coded protein of Klebsiella pneumoniae.

    OpenAIRE

    Stacey, G.; Zhu, J.; Shah, V K; Shen, S C; Brill, W J

    1982-01-01

    A single mutation, nifC1005 (Jin et al. Sci. Sin. 23:108-118, 1980), located between nifH and nifJ in the nif cluster of Klebsiella pneumoniae, genetically complemented mutations in each of the 17 known nif genes. This suggested that the mutation is located in a new nif gene. We showed by complementation analyses that only 3 of 12 nifJ mutations tested were complemented by nifC1005. Nitrogenase activity in cell extracts of the mutant with nifC1005 as well as NifJ- mutants was stimulated by th...

  18. Simultaneous differential detection of Chlamydophila abortus, Chlamydophila pecorum and Coxiella burnetii from aborted ruminant's clinical samples using multiplex PCR

    Directory of Open Access Journals (Sweden)

    Rodolakis Annie

    2009-07-01

    Full Text Available Abstract Background Chlamydiosis and Q fever, two zoonosis, are important causes of ruminants' abortion around the world. They are caused respectively by strictly intracellular and Gram negative bacterium Chlamydophila abortus (Cp. abortus and Coxiella burnetii (C. burnetii. Chlamydophila pecorum (Cp. pecorum is commonly isolated from the digestive tract of clinically inconspicuous ruminants but the abortive and zoonotic impact of this bacterium is still unknown because Cp. pecorum is rarely suspected in abortion cases of small ruminants. We have developed a multiplex PCR (m-PCR for rapid simultaneous differential detection of Cp. abortus, Cp. pecorum and C. burnetii in clinical samples taken from infected animals. Results Specific PCR primers were designed and a sensitive and specific m-PCR was developed to detect simultaneously, in one tube reaction, three specific fragments of 821, 526 and 687-bp long for Cp. abortus, Cp. pecorum and C. burnetii respectively. This m-PCR assay was performed on 253 clinical samples taken from infected ruminant's flocks that have showed problems of abortion diseases. Thus, 67 samples were infected by either one of the three pathogens: 16 (13 vaginal swabs and 3 placentas were positive for Cp. abortus, 2 were positive for Cp. pecorum (1 vaginal swab and 1 placenta and 49 samples (33 vaginal swabs, 11 raw milks, 4 faeces and 1 placenta were positive for C. burnetii. Two vaginal swabs were m-PCR positive of both Cp. abortus and C. burnetii and none of the tested samples was shown to be infected simultaneously with the three pathogens. Conclusion We have successfully developed a rapid multiplex PCR that can detect and differentiate Cp. abortus, Cp. pecorum and C. burnetii; with a good sensitivity and specificity. The diagnosis of chlamydiosis and Q fever may be greatly simplified and performed at low cost. In addition, the improvement in diagnostic techniques will enhance our knowledge regarding the prevalence and

  19. Identification and characterisation of coding tandem repeat variants in incA gene of Chlamydophila pecorum.

    Science.gov (United States)

    Yousef Mohamad, Khalil; Rekiki, Abdessalem; Myers, Garry; Bavoil, Patrik M; Rodolakis, Annie

    2008-01-01

    Bacteria of the family Chlamydiaceae are obligate intracellular pathogens of human and animals. Chlamydophila pecorum is associated with different pathological conditions in ruminants, swine and koala. To characterize a coding tandem repeat (CTR) identified at the 3' end of incA gene of C. pecorum, 51 strains of different chlamydial species were examined. The CTR were observed in 18 of 18 tested C. pecorum isolates including symptomatic and asymptomatic animals from diverse geographical origins. The CTR were also found in two strains of C. abortus respectively isolated from faeces from a healthy ewe and from a goat belonging to asymptomatic herds, but were absent in C. abortus strains isolated from clinical disease specimens, and in tested strains of C. psittaci, C. caviae, C. felis and C. trachomatis. The number of CTR repeats is variable and encode several motifs that are rich in alanine and proline. The CTR-derived variable structure of incA, which encode the Chlamydiaceae-specific type III secreted inclusion membrane protein, IncA, may be involved in the adaptation of C. pecorum to its environment by allowing it to persist in the host cell.

  20. Pneumonia (image)

    Science.gov (United States)

    Pneumonia is an inflammation of the lungs caused by an infection. Many different organisms can cause it, including bacteria, viruses, and fungi. Pneumonia is a common illness that affects millions of ...

  1. Understanding Pneumonia

    Science.gov (United States)

    ... Pneumonia Awareness Campaign, on World Pneumonia Day Blog: Yoga, Tai Chi and Your Lungs: The Benefits of ... number of items"); $("#local_list_xml").quickPagination(); }, error: function() { console.log("An error occurred while processing XML ...

  2. Using CF0218-ELISA to distinguish Chlamydophila felis-infected cats from vaccinated and uninfected domestic cats.

    Science.gov (United States)

    Ohya, Kenji; Okuda, Hideko; Maeda, Sadatoshi; Yamaguchi, Tsuyoshi; Fukushi, Hideto

    2010-12-15

    Chlamydophila felis is a causative agent of acute and chronic conjunctivitis and pneumonia in cats. Cats can be vaccinated with killed or attenuated C. felis. However, current serodiagnostics cannot distinguish these cats from naturally infected cats. This causes difficulty of early diagnosis and seroepidemiological survey for C. felis. We previously reported that C. felis CF0218 can be used as a C. felis-infection-specific diagnostic antigen in experimentally infected and/or vaccinated cats. In this study, we evaluated an enzyme-linked immunosorbent assay using recombinant CF0218 as antigen (CF0218-ELISA) to detect anti-C. felis antibody in 714 sera of domestic cats whose histories of vaccination against C. felis are known. The 44 vaccinated cats were 93% negative using CF0218-ELISA; half of these scored positive by immunofluorescence assay (IFA) using C. felis-infected cells as antigen. The 670 non-vaccinated cats had CF0218-ELISA positivity rates that were statistically in agreement with IFA (18% vs. 21%). These results show that CF0218, which was identified as a C. felis-infection-specific antigen, is a useful serodiagnostic antigen to distinguish naturally C. felis-infected cats from vaccinated and non-infected cats.

  3. Classification of idiopathic interstitial pneumonias using anti–myxovirus resistance-protein 1 autoantibody

    Science.gov (United States)

    Hamano, Yoshimasa; Kida, Hiroshi; Ihara, Shoichi; Murakami, Akihiro; Yanagawa, Masahiro; Ueda, Ken; Honda, Osamu; Tripathi, Lokesh P.; Arai, Toru; Hirose, Masaki; Hamasaki, Toshimitsu; Yano, Yukihiro; Kimura, Tetsuya; Kato, Yasuhiro; Takamatsu, Hyota; Otsuka, Tomoyuki; Minami, Toshiyuki; Hirata, Haruhiko; Inoue, Koji; Nagatomo, Izumi; Takeda, Yoshito; Mori, Masahide; Nishikawa, Hiroyoshi; Mizuguchi, Kenji; Kijima, Takashi; Kitaichi, Masanori; Tomiyama, Noriyuki; Inoue, Yoshikazu; Kumanogoh, Atsushi

    2017-01-01

    Chronic fibrosing idiopathic interstitial pneumonia (IIP) can be divided into two main types: idiopathic pulmonary fibrosis (IPF), a steroid-resistant and progressive disease with a median survival of 2–3 years, and idiopathic non-specific interstitial pneumonia (INSIP), a steroid-sensitive and non-progressive autoimmune disease. Although the clinical courses of these two diseases differ, they may be difficult to distinguish at diagnosis. We performed a comprehensive analysis of serum autoantibodies from patients definitively diagnosed with IPF, INSIP, autoimmune pulmonary alveolar proteinosis, and sarcoidosis. We identified disease-specific autoantibodies and enriched KEGG pathways unique to each disease, and demonstrated that IPF and INSIP are serologically distinct. Furthermore, we discovered a new INSIP-specific autoantibody, anti–myxovirus resistance-1 (MX1) autoantibody. Patients positive for anti-MX1 autoantibody constituted 17.5% of all cases of chronic fibrosing IIPs. Notably, patients rarely simultaneously carried the anti-MX1 autoantibody and the anti–aminoacyl-transfer RNA synthetase autoantibody, which is common in chronic fibrosing IIPs. Because MX1 is one of the most important interferon-inducible anti-viral genes, we have not only identified a new diagnostic autoantibody of INSIP but also obtained new insight into the pathology of INSIP, which may be associated with viral infection and autoimmunity. PMID:28230086

  4. PENICILLIN-BINDING PROTEIN 2X OF STREPTOCOCCUS-PNEUMONIAE - EXPRESSION IN ESCHERICHIA-COLI AND PURIFICATION OF A SOLUBLE ENZYMATICALLY ACTIVE DERIVATIVE

    NARCIS (Netherlands)

    LAIBLE, G; KECK, W; LURZ, R; MOTTL, H; FRERE, JM; JAMIN, M; HAKENBECK, R

    1992-01-01

    A 2.5-kb DNA fragment including the structural gene coding for the penicillin-binding protein 2x (PBP 2x) of Streptococcus pneumoniae has been cloned into the vector pJDC9 and expressed in Escherichia coli. Mapping of RNA polymerase binding sites by electron microscopy indicated that the pbpX promot

  5. Seroprevalence and risk factors associated with Chlamydophila spp. infection in ewes in the northeast of Algeria.

    Science.gov (United States)

    Hireche, Sana; Bouaziz, Omar; Djenna, Djahida; Boussena, Sabrina; Aimeur, Rachida; Kabouia, Rachid; Bererhi, El Hacène

    2014-02-01

    A cross-sectional study was carried out to estimate prevalence of Chlamydophila spp. antibodies and to investigate risk factors associated with chlamydial infection in 552 ewes between March 2011 and January 2012 in the province of Constantine. Anti-Chlamydophila antibodies were detected using an indirect enzyme-linked immunosorbent assay kit in 24.5% of examined sera. Of the herds, 70.4% had at least one seropositive animal. A pretested structured questionnaire was administered in order to collect information on individual animal health and herd management practices. Chi-square analysis and multivariable logistic regression model were used to identify risk factors related to Chlamydophila seropositivity. Univariable analysis revealed 17 variables with p Chlamydophila spp. seropositivity. Moreover, occurrence of stillbirth problem (OR = 3.682, 95% CI (OR) = 1.825; 7.430) and 5-10% mortality rate in young lambs (OR = 2.584, 95% CI (OR) = 1.058; 6.310) were significantly associated with seropositivity to Chlamydophila spp. On the other hand, availability of veterinary service was identified as a protective factor (OR = 0.161, 95% CI (OR) = 0.051; 0.511).

  6. Crystallization and preliminary X-ray diffraction studies of choline-binding protein F from Streptococcus pneumoniae

    Energy Technology Data Exchange (ETDEWEB)

    Molina, Rafael [Grupo de Cristalografía Macromolecular y Biología Estructural, Instituto Química Física Rocasolano, CSIC, Serrano 119, 28006 Madrid (Spain); González, Ana; Moscoso, Miriam; García, Pedro [Departamento de Microbiología Molecular, Centro de Investigaciones Biológicas, CSIC, Ramiro de Maeztu 9, 28040 Madrid (Spain); Stelter, Meike; Kahn, Richard [Institut de Biologie Structurale J.-P. Ebel CEA CNRS UJF, Laboratoire de Cristallographie Macromoléculaire, 41 Rue Jules Horowitz, 38027 Grenoble CEDEX 1 (France); Hermoso, Juan A., E-mail: xjuan@iqfr.csic.es [Grupo de Cristalografía Macromolecular y Biología Estructural, Instituto Química Física Rocasolano, CSIC, Serrano 119, 28006 Madrid (Spain)

    2007-09-01

    The modular choline-binding protein F (CbpF) from S. pneumoniae has been crystallized by the hanging-drop vapour-diffusion method. A SAD data set from a gadolinium-complex derivative has been collected to 2.1 Å resolution. Choline-binding protein F (CbpF) is a modular protein that is bound to the pneumococcal cell wall through noncovalent interactions with choline moieties of the bacterial teichoic and lipoteichoic acids. Despite being one of the more abundant proteins on the surface, along with the murein hydrolases LytA, LytB, LytC and Pce, its function is still unknown. CbpF has been crystallized using the hanging-drop vapour-diffusion method at 291 K. Diffraction-quality orthorhombic crystals belong to space group P2{sub 1}2{sub 1}2, with unit-cell parameters a = 49.13, b = 114.94, c = 75.69 Å. A SAD data set from a Gd-HPDO3A-derivatized CbpF crystal was collected to 2.1 Å resolution at the gadolinium L{sub III} absorption edge using synchrotron radiation.

  7. Molecular detection of Chlamydophila felis in ocular discharge of domestic cats in Tehran & Isfahan

    Directory of Open Access Journals (Sweden)

    Hassan Momtaz

    2014-04-01

    Full Text Available Introduction: Chlamydophila felis is one of the most common pathogens isolated from ocular discharges in domestic cats. The aim of the present study was to detect this microorganism in ocular discharges collected from domestic cats in Tehran and Isfahan cities Materials and methods: From a total of 224 domestic cats, ocular swabs were collected and DNA was extracted. Samples were studied using the Polymerase Chain Reaction method and their products were analyzed using agarose gel. Results: From a total of 224 samples studied, 40 samples (17.85% were infected with Chlamydophila felis which were diagnosed as positive in PCR test. Discussion and conclusion: According to the difficulties and time-consuming process of isolation of Chlamydophila felis in laboratory culture media, the PCR technique has been suggested as a rapid and safe method for detection of this organism in clinical samples.

  8. Detection of the ompA gene of Chlamydophila pecorum in captive birds in Argentina.

    Science.gov (United States)

    Frutos, María C; Venezuela, Fernando; Kiguen, Ximena; Ré, Viviana; Cuffini, Cecilia

    2012-01-01

    Bacteria belonging to the family Chlamydiaceae cause a broad spectrum of diseases in a wide range of hosts, including humans, other mammals and birds. However, very little is known about chlamydial infections in birds in our region. In the present study, we examined 28 clinically normal birds in illegal captivity that were confiscated in the province of C6rdoba, Argentina. The objective was to detect Chlamydophila spp. in cloacal swabs by genetic analysis of the ompA gene. Nested-PCR of the ompA gene identified five samples as Chlamydophila pecorum and the sequence analysis demonstrated the presence of the ompA gene of C. pecorum in these birds. On the other hand, Chlamydophila psittaci was not detected. These birds could be either asymptomatic reservoirs or subclinical carriers of C. pecorum. This is the first report of the detection of C. pecorum in Argentina.

  9. Increased prevalence of Chlamydophila DNA in post-mortem brain frontal cortex from patients with schizophrenia.

    Science.gov (United States)

    Fellerhoff, Barbara; Wank, Rudolf

    2011-07-01

    Infection can initiate symptoms of mental illness. It has been shown previously that Chlamydophila DNA is present six times more often in the blood of patients with schizophrenia than in the blood of control individuals. Monocytes, the main targets of Chlamydiaceae infection, are microglia precursors. We identified Chlamydiaceae infection using blinded brain DNA samples derived from the frontal cortex. Using PCR and sequence analysis, we found Chlamydophila DNA to be four times greater in patients with schizophrenia than in controls (schizophrenia: N=34, microbial DNA frequency 23.5%; controls: N=35, microbial DNA frequency 5.7%; P=0.045, OR=5.08). Persistent Chlamydophila-infected microglia or neuronal cells may impair neuronal circuits and thus be a mechanism for causing psychiatric illness in these patients.

  10. Clinical and economic burden of community-acquired pneumonia amongst adults in the Asia-Pacific region.

    Science.gov (United States)

    Song, Jae-Hoon; Thamlikitkul, Visanu; Hsueh, Po-Ren

    2011-08-01

    Community-acquired pneumonia (CAP) is an important cause of mortality and morbidity amongst adults in the Asia-Pacific region. Literature published between 1990 and May 2010 on the clinical and economic burden of CAP amongst adults in this region was reviewed. CAP is a significant health burden with significant economic impact in this region. Chronic obstructive pulmonary disease, cardiovascular disease, diabetes mellitus and advanced age were risk factors for CAP. Aetiological agents included Streptococcus pneumoniae, Klebsiella pneumoniae, Gram-negative bacteria, Mycobacterium tuberculosis, Burkholderia pseudomallei, Staphylococcus aureus and atypical pathogens (Mycoplasma pneumoniae, Chlamydophila pneumoniae and Legionella spp.), with important differences in the prevalence of these pathogens within the region. Antibiotic resistance was significant but was not linked to excess mortality. Aetiological pathogens remained susceptible to newer antimicrobial agents. Rational antibiotic use is essential for preventing resistance, and increased surveillance is required to identify future trends in incidence and aetiology and to drive treatment and prevention strategies.

  11. 鹦鹉热嗜衣原体基因组学的研究进展%Research progress on genomics of Chlamydophila psittaci

    Institute of Scientific and Technical Information of China (English)

    黎知青

    2012-01-01

    目前,7株鹦鹉热嗜衣原体全基因组己测序完整,揭示了不同株之间的相同性与差异性.主要外膜蛋白基因、多形态膜蛋白多基因家族、Ⅲ型分泌系统基因和包涵体膜蛋白基因是鹦鹉热嗜表原体(Chlamydophila psittaci,Cps)研究的重点.对多个种株Cps基因组完整核苷酸序列测定的研究,将有助于进一步了解Cps的致病机制,寻找更好的诊断方法和防治措施,预防和控制Cps的流行.%At present, the whole genomes of seven strains Chlamydophila psittaci (Cp) have been completely se-quenced and it reveals the similarities and differences among each other. Major outer membrane protein genes, the polymorphic membrane protein multiple genes family, the type Ⅲ secretion system genes, and the inclusion membrane protein genes are the focuses of study on Chlamydophila psittaci. The studies on genome integrity nucleotide sequencing among multiple strains of Cps will help for further understanding of the pathogenesis of Cps, searching for better diagnostic methods and prevention measures, and preventing and controlling the prevalence of Cps.

  12. Co-administration of the polysaccharide of Lycium barbarum with DNA vaccine of Chlamydophila abortus augments protection.

    Science.gov (United States)

    Ling, Yong; Li, Shaowen; Yang, Junjing; Yuan, Jilei; He, Cheng

    2011-01-01

    Lycium barbarum polysaccharides (LBP) can stimulate moderate immune responses therefore could potentially be used as a substitute for oil adjuvants in veterinary vaccines. In the present study, it was shown that the isolated active component of LBP3a, combined with a DNA vaccine encoding the major outer membrane protein (MOMP) of Chlamydophila abortus, induced protection in mice against challenge. Sixty BALB/c mice were randomly assigned to 5 groups. Sub-fractions of polysaccharide LBP3a, at 12.5, 25 and 50 mg/kg concentrations, respectively, were mixed with a pCI-neo::MOMP (pMOMP) vaccine. Mice administrated with pCI-neo + LBP3a were served as a control. All mice were inoculated at day 0, 14, and 28, and challenged on day 44. The effects of LBp3a on serum antibody levels, in vitro lymphocyte proliferation, the activity of interleaukin-2 (IL-2), interferon-γ (IFN-γ), tumor necrosis factor α(TNF-α)and chlamydia clearance were determined. A combination of DNA vaccine and LBP3a induced significantly higher antibody levels in mice, higher T cell proliferation and higher levels of IFN-γ and IL-2. Mice immunized with DNA and LBP3a also showed significantly higher levels of chlamydia clearance in mice spleens and a greater Th1 immune response. The immunoenhancement induced by 25 mg/kg LBP3a is more effective than that induced by a 12.5 and 50 mg/kg. This implies that LBP3a at 25 mg/kg has a high potential to be used as an effective adjuvant with a DNA vaccine against swine Chlamydophila abortus.

  13. Unique nonstructural proteins of Pneumonia Virus of Mice (PVM) promote degradation of interferon (IFN) pathway components and IFN-stimulated gene proteins.

    Science.gov (United States)

    Dhar, Jayeeta; Barik, Sailen

    2016-12-01

    Pneumonia Virus of Mice (PVM) is the only virus that shares the Pneumovirus genus of the Paramyxoviridae family with Respiratory Syncytial Virus (RSV). A deadly mouse pathogen, PVM has the potential to serve as a robust animal model of RSV infection, since human RSV does not fully replicate the human pathology in mice. Like RSV, PVM also encodes two nonstructural proteins that have been implicated to suppress the IFN pathway, but surprisingly, they exhibit no sequence similarity with their RSV equivalents. The molecular mechanism of PVM NS function, therefore, remains unknown. Here, we show that recombinant PVM NS proteins degrade the mouse counterparts of the IFN pathway components. Proteasomal degradation appears to be mediated by ubiquitination promoted by PVM NS proteins. Interestingly, NS proteins of PVM lowered the levels of several ISG (IFN-stimulated gene) proteins as well. These results provide a molecular foundation for the mechanisms by which PVM efficiently subverts the IFN response of the murine cell. They also reveal that in spite of their high sequence dissimilarity, the two pneumoviral NS proteins are functionally and mechanistically similar.

  14. Penicillin-binding protein 3 of Streptococcus pneumoniae and its application in screening of β-lactams in milk.

    Science.gov (United States)

    Zhang, Jing; Wang, Zhanhui; Wen, Kai; Liang, Xiao; Shen, Jianzhong

    2013-11-15

    The soluble form of penicillin-binding protein 3 (sPBP3(∗)) from Streptococcus pneumoniae was expressed in Escherichia coli as a six-histidine fusion protein. The protein was purified and used to develop a microplate assay in direct competitive format for the detection of penicillins and cephalosporins in milk. The assay was based on competitive inhibition of the binding of horseradish peroxidase-labeled ampicillin (HRP-Amp) to the sPBP3(∗) by free β-lactam antibiotics in milk. Under optimized conditions, most of the β-lactam antibiotics (11 penicillins and 16 cephalosporins) could be detected at concentrations corresponding to the maximum residue limits (MRLs) set by the European Union. Analysis of spiked milk samples showed that acceptable recoveries ranged from 74.06 to 106.31% in skimmed milk and from 63.97 to 107.26% in whole milk, with coefficients of variation (CVs) less than 16%. With the high sensitivity and wide-range affinities to penicillins and cephalosporins, the developed assay based on sPBP3(∗) exhibited the potential to be a screening assay for fast detection of β-lactam antibiotics in milk.

  15. Amniotic and allantoic fluids from experimentally infected sheep contain immunoglobulin specific for Chlamydophila abortus.

    Science.gov (United States)

    Marques, Patricia X; O'Donovan, James; Souda, Puneet; Gutierrez, Jorge; Williams, Erin J; Worrall, Sheila; McElroy, Maire; Proctor, Aisling; Brady, Colm; Sammin, Donal; Basset, Hugh; Whitelegge, Julian P; Markey, Bryan K; Nally, Jarlath E

    2011-03-15

    Chlamydophila abortus, the aetiological agent of enzootic abortion of ewes (EAE), replicates in trophoblast cells leading to their destruction and dissemination of the bacterium to foetal organs. To further understand the pathogenesis of EAE, amniotic and allantoic fluids were collected from experimentally infected pregnant ewes at 30 (7 samples from each fluid), 35 (8 samples from each fluid), 40 (10 samples from each fluid) and 43 (6 amniotic fluids and 7 allantoic fluids) days post-infection to determine pathogen numbers and other markers of infection. Whilst experimentally infected ewes had characteristic placental lesions, only two amniotic and seven allantoic fluid samples were positive for C. abortus by real-time PCR. In contrast, all amniotic and allantoic fluids were positive for immunoglobulin. Immunoglobulins were generally detected earlier in allantoic fluid than in amniotic fluid and the numbers of samples containing immunoglobulins increased as infection progressed. IgG in amniotic and allantoic fluids was shown to be specific for C. abortus, and reacted with the major outer membrane proteins, polymorphic outer membrane protein and macrophage infectivity potentiator protein. A comparison of two-dimensional immunoblots using purified IgG from the allantoic fluid, amniotic fluid, ewe serum and foetal serum of a C. abortus infected animal at 40 days post infection indicated a pattern of reactivity intermediate between that of the ewe serum and the foetal serum. Results suggest that a maternal source of immunoglobulin is predominant at 30 days post-infection but that foetal derived antibodies may be contributed at a later stage.

  16. Host alpha-adducin is redistributed and localized to the inclusion membrane in chlamydia- and chlamydophila-infected cells.

    Science.gov (United States)

    Chu, Hencelyn G; Weeks, Sara K; Gilligan, Diana M; Rockey, Daniel D

    2008-12-01

    A large-scale analysis of proteins involved in host-cell signalling pathways was performed using chlamydia-infected murine cells in order to identify host proteins that are differentially activated or localized following infection. Two proteins whose distribution was altered in Chlamydia trachomatis-infected cells relative to mock-infected cells were the actin-binding protein adducin and the regulatory kinase Raf-1. Immunoblot analysis with antibodies to both phosphorylated and non-phosphorylated forms of these proteins demonstrated that the abundance of each protein was markedly reduced in the cytosolic fraction of C. trachomatis- and Chlamydophila caviae-infected cells, but the total cellular protein abundance remained unaffected by infection. Fluorescence microscopy of chlamydia-infected cells using anti-alpha-adducin antibodies demonstrated labelling at or near the chlamydial inclusion membrane. Treatment of infected cells with nocodazole or cytochalasin D did not affect alpha-adducin that was localized to the margins of the inclusion. The demonstration of alpha-adducin and Raf-1 redistribution within cells infected by different chlamydiae provides novel opportunities for analysis of host-pathogen interactions in this system.

  17. Clinical value of recombinant protein of chlamydial protease-like activity factor from Chlamydia pneumoniae in serodiagnosis%肺炎嗜衣原体蛋白酶样活性因子重组蛋白的血清学诊断价值初探

    Institute of Scientific and Technical Information of China (English)

    郑江花; 吴移谋; 丁滔; 陈丽丽; 莫小辉; 李忠玉

    2008-01-01

    Objective To clone and express the immunodominant region gene of the chlamydial protease-like activity factor (CPAF) from Chlamydophila pneumoniae (Cpn), and then evaluate the clinical value of the recombinant protein in serodiagnosis.Methods The pGEX6p-2/CPAFm recombinant plasmid was constructed, and then expression of recombinant protein was induced. The expressed recombinant protein was identified by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). New Zealand white rabbits were immunized with the purified protein;indirect ELISA was applied to measuring its immunogenicity;and its immunoreactivity was analyzed by reacting with C.pneumoniae-specific IgG antibody in sera. Cpn IgG and Chlamydozoa trachomatis positive sera were detected with indirect ELISA.Results The recombinant protein with relative molecular weight about 51.3×103 was expressed and purified effectively. Western blot analysis proved that the recombinant protein could react with human anti-C.pneumoniae IgG specifically. The titer of 000. 40 cases of C.pneumuniae IgG reference sera were detected with indirect ELISA, and the concordance rate of negative and positive results were both 100.0% (20/20). C.pneumoniae-specific IgG antibody in 300 clinical serum specimens was detected with indirect ELISA and microimmunofluorescence (MIF), and the concordance rate between the two methods was 98.3%. No cross reaction against C.trachomatis was found with indirect ELISA as anti-C.trachomatis positive sera were detected.Conclusion The expressed recombinant protein of CPAF immunodominant region gene from C.pneumoniae shows excellent immunocompetence. It is suitable for preparation of serodiagnostic ELISA reagent for C.pneumoniae infection.%目的 克隆、表达肺炎嗜衣原体(Cpn)蛋白酶样活性因子(CPAF)的免疫优势区基因(CPAFm),初步评价其在血清学诊断中的应用价值.方法 构建pGEX6p-2/CPAFm重组质粒,诱导表达重组蛋白,十二烷基磺酸钠-聚丙烯

  18. Epidemiological study of Chlamydophila psittaci in pet birds in Croatia

    Directory of Open Access Journals (Sweden)

    Križek I.

    2012-01-01

    Full Text Available A total of 411 samples from birds of different species originating from all counties of the Republic of Croatia have been tested for the presence of Chlamydophila psittaci. The sampling was conducted in pet stores, breeders' aviaries, in a specialized bird clinic and in zoos. The testing included 177 parrots, 169 pigeons, 58 canaries and 7 finches. For the detection of specific C. psittaci antigen a commercial ELISA kit was used- IDEIATM PCE Chlamydia (DAKO Cytomation Ltd., United Kingdom. The samples that were non-specifically positive or doubtful in the ELISA test (a total of 26 samples were analyzed also by means of polymerase chain reaction (PCR. Diagnostic ELISA method found a total of 17.03% birds positive for chlamydiosis, and after additional testing by PCR a total of 12.65% positive ones were found. According to bird species, the most frequently positive ones were canaries and pigeons (15.52% and 13.02%, and according to the sampling location most of the positive birds were found in pet stores (16.52%, but a high percentage of positive samples were also found in breeders’ aviaries (11.76%. The average positive result for chlamydiosis in 12.65% of tested birds is alarming and it confirms the importance of monitoring bird health and of prescribed legal regulations when it comes to chlamydial diseases, as well as education of persons involved in breeding, keeping or selling birds.

  19. NCBI nr-aa BLAST: CBRC-RMAC-03-0003 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-RMAC-03-0003 ref|NP_300247.1| hypothetical protein CPj0188 [Chlamydophila pneu...moniae J138] dbj|BAA98398.1| CT132 hypothetical protein [Chlamydophila pneumoniae J138] NP_300247.1 0.013 20% ...

  20. NCBI nr-aa BLAST: CBRC-RMAC-03-0003 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-RMAC-03-0003 ref|NP_445122.1| hypothetical protein CP0579 [Chlamydophila pneum...oniae AR39] gb|AAF38397.1| conserved hypothetical protein [Chlamydophila pneumoniae AR39] NP_445122.1 0.018 20% ...

  1. Photodynamics of blue-light-regulated phosphodiesterase BlrP1 protein from Klebsiella pneumoniae and its photoreceptor BLUF domain

    Energy Technology Data Exchange (ETDEWEB)

    Tyagi, A. [Institut II - Experimentelle und Angewandte Physik, Universitaet Regensburg, Universitaetstrasse 31, D-93053 Regensburg (Germany); Penzkofer, A. [Institut II - Experimentelle und Angewandte Physik, Universitaet Regensburg, Universitaetstrasse 31, D-93053 Regensburg (Germany)], E-mail: alfons.penzkofer@physik.uni-regensburg.de; Griese, J.; Schlichting, I. [Max-Planck-Institut fuer medizinische Forschung, Abteilung Biomolekulare Mechanismen, Jahnstrasse 29, D-69120 Heidelberg (Germany); Kirienko, Natalia V.; Gomelsky, Mark [Department of Molecular Biology, University of Wyoming, Laramie, Wyoming 82071 (United States)

    2008-12-10

    The BlrP1 protein from the enteric bacterium Klebsiella pneumoniae consists of a BLUF and an EAL domain and may activate c-di-GMP phosphodiesterase by blue-light. The full-length protein, BlrP1, and its BLUF domain, BlrP1{sub B}LUF, are characterized by optical absorption and emission spectroscopy. The cofactor FAD in its oxidized redox state (FAD{sub ox}) is brought from the dark-adapted receptor state to the 10-nm red-shifted putative signalling state by violet light exposure. The recovery to the receptor state occurs with a time constant of about 1 min. The quantum yield of signalling state formation is about 0.17 for BlrP1{sub B}LUF and about 0.08 for BlrP1. The fluorescence efficiency of the FAD{sub ox} cofactor is small due to photo-induced reductive electron transfer. Prolonged light exposure converts FAD{sub ox} in the signalling state to the fully reduced hydroquinone form FAD{sub red}H{sup -} and causes low-efficient chromophore release with subsequent photo-degradation. The photo-cycle and photo-reduction dynamics in the receptor state and in the signalling state are discussed.

  2. Photodynamics of blue-light-regulated phosphodiesterase BlrP1 protein from Klebsiella pneumoniae and its photoreceptor BLUF domain

    Science.gov (United States)

    Tyagi, A.; Penzkofer, A.; Griese, J.; Schlichting, I.; Kirienko, Natalia V.; Gomelsky, Mark

    2008-12-01

    The BlrP1 protein from the enteric bacterium Klebsiella pneumoniae consists of a BLUF and an EAL domain and may activate c-di-GMP phosphodiesterase by blue-light. The full-length protein, BlrP1, and its BLUF domain, BlrP1_BLUF, are characterized by optical absorption and emission spectroscopy. The cofactor FAD in its oxidized redox state (FAD ox) is brought from the dark-adapted receptor state to the 10-nm red-shifted putative signalling state by violet light exposure. The recovery to the receptor state occurs with a time constant of about 1 min. The quantum yield of signalling state formation is about 0.17 for BlrP1_BLUF and about 0.08 for BlrP1. The fluorescence efficiency of the FAD ox cofactor is small due to photo-induced reductive electron transfer. Prolonged light exposure converts FAD ox in the signalling state to the fully reduced hydroquinone form FAD redH - and causes low-efficient chromophore release with subsequent photo-degradation. The photo-cycle and photo-reduction dynamics in the receptor state and in the signalling state are discussed.

  3. The pavA gene of Streptococcus pneumoniae encodes a fibronectin-binding protein that is essential for virulence

    NARCIS (Netherlands)

    Holmes, AR; McNab, R; Millsap, KW; Rohde, M; Hammerschmidt, S; Mawdsley, JL; Jenkinson, HF

    2001-01-01

    Streptococcus pneumoniae colonizes the nasopharynx in up to 40% of healthy subjects, and is a leading cause of middle ear infections (otitis media), meningitis and pneumonia. Pneumococci adhere to glycosidic receptors on epithelial cells and to immobilized fibronectin, but the bacterial adhesins med

  4. Control of cell division in Streptococcus pneumoniae by the conserved Ser/Thr protein kinase StkP

    NARCIS (Netherlands)

    Beilharz, Katrin; Nováková, Linda; Fadda, Daniela; Branny, Pavel; Massidda, Orietta; Veening, Jan-Willem

    2012-01-01

    How the human pathogen Streptococcus pneumoniae coordinates cell-wall synthesis during growth and division to achieve its characteristic oval shape is poorly understood. The conserved eukaryotic-type Ser/Thr kinase of S. pneumoniae, StkP, previously was reported to phosphorylate the cell-division

  5. Prediction of Genomic Islands in Three Bacterial Pathogens of Pneumonia

    Directory of Open Access Journals (Sweden)

    Wen Wei

    2012-03-01

    Full Text Available Pneumonia is one kind of common infectious disease, which is usually caused by bacteria, viruses, or fungi. In this paper, we predicted genomic islands in three bacterial pathogens of pneumonia. They are Chlamydophila pneumoniae, Mycoplasma pneumoniae and Streptococcus pneumoniae, respectively. For each pathogen, one clinical strain is involved. After implementing the cumulative GC profile combined with h and BCN index, eight genomic islands are found in three pathogens. Among them, six genomic islands are found to have mobility elements, which constitute a kind of conserved character of genomic islands, and this introduces the possibility that they are genuine genomic islands. The present results show that the cumulative GC profile when combined with h and BCN indexes is a good method for predicting genomic islands in bacteria and it has lower false positive rate than the SIGI method. Specially, three genomic islands are found to contain clusters of genes coding for production of virulence factors and this is useful for research into the pathogenicity of these pathogens and helpful for the treatment of diseases caused by them.

  6. An outbreak of psittacosis due to Chlamydophila psittaci genotype A in a veterinary teaching hospital

    NARCIS (Netherlands)

    E.R. Heddema; E.J. van Hannen; B. Duim; B.M. de Jongh; J.A. Kaan; R. van Kessel; J.T. Lumeij; C.E. Visser; C.M.J.E. Vandenbroucke-Grauls

    2006-01-01

    An outbreak of psittacosis in a veterinary teaching hospital was recognized in December 2004. Outbreak management was instituted to evaluate the extent of the outbreak and to determine the avian source. Real-time PCR, serologic testing and sequencing of the ompA gene of Chlamydophila psittaci were p

  7. Reproductive disorders induced by Chlamydophila spp. infections in an italian mediterranean buffalo (bubalus bubalis herd

    Directory of Open Access Journals (Sweden)

    M. Corrente

    2010-02-01

    Full Text Available The Italian Mediterranean Buffalo (Bubalus bubalis has low fecundity and high incidence of abortion. Several studies have associated reproductive failure of water buffalo with viral infections but there is limited information on the role of chlamydial infections. To investigate the presence and the role of Chlamydiaceae in water buffalo a retrospective study was performed in a farm where, in the arch of 11 months, the pregnant heifers suffered an abortion rate of 36.8% in the 3rd and 5th month of pregnancy. Antibodies to Chlamydiaceae were detected in 57% of the aborted cows, while the rate of positivity was 0% in overtly healthy cows used as control. By a PCR assay 3 of 14 vaginal swabs from aborted animals tested positive for Chlamydophila agents and, additionally, 3 out of 7 aborted foetuses tested positive for Chlamydophila spp., with two being co-infections by Cp. abortus and Cp. pecorum and one being characterised as Cp. abortus. The presence of anti-Chlamydiaceae antibodies in 57% of the aborted animals and the detection of Chlamydophila agents in foetal organs and in vaginal swabs is consistent with the history of abortions (P<0.002 observed in the herd and may suggest a pathogenic role by Chlamydophila spp. in water buffalo.

  8. Survey of clustered regularly interspaced short palindromic repeats and their associated Cas proteins (CRISPR/Cas) systems in multiple sequenced strains of Klebsiella pneumoniae.

    Science.gov (United States)

    Ostria-Hernández, Martha Lorena; Sánchez-Vallejo, Carlos Javier; Ibarra, J Antonio; Castro-Escarpulli, Graciela

    2015-08-04

    In recent years the emergence of multidrug resistant Klebsiella pneumoniae strains has been an increasingly common event. This opportunistic species is one of the five main bacterial pathogens that cause hospital infections worldwide and multidrug resistance has been associated with the presence of high molecular weight plasmids. Plasmids are generally acquired through horizontal transfer and therefore is possible that systems that prevent the entry of foreign genetic material are inactive or absent. One of these systems is CRISPR/Cas. However, little is known regarding the clustered regularly interspaced short palindromic repeats and their associated Cas proteins (CRISPR/Cas) system in K. pneumoniae. The adaptive immune system CRISPR/Cas has been shown to limit the entry of foreign genetic elements into bacterial organisms and in some bacteria it has been shown to be involved in regulation of virulence genes. Thus in this work we used bioinformatics tools to determine the presence or absence of CRISPR/Cas systems in available K. pneumoniae genomes. The complete CRISPR/Cas system was identified in two out of the eight complete K. pneumoniae genomes sequences and in four out of the 44 available draft genomes sequences. The cas genes in these strains comprises eight cas genes similar to those found in Escherichia coli, suggesting they belong to the type I-E group, although their arrangement is slightly different. As for the CRISPR sequences, the average lengths of the direct repeats and spacers were 29 and 33 bp, respectively. BLAST searches demonstrated that 38 of the 116 spacer sequences (33%) are significantly similar to either plasmid, phage or genome sequences, while the remaining 78 sequences (67%) showed no significant similarity to other sequences. The region where the CRISPR/Cas systems were located is the same in all the Klebsiella genomes containing it, it has a syntenic architecture, and is located among genes encoding for proteins likely involved in

  9. Streptococcus pneumoniae Cell-Wall-Localized Phosphoenolpyruvate Protein Phosphotransferase Can Function as an Adhesin: Identification of Its Host Target Molecules and Evaluation of Its Potential as a Vaccine.

    Directory of Open Access Journals (Sweden)

    Yaffa Mizrachi Nebenzahl

    Full Text Available In Streptococcus pneumonia, phosphoenolpyruvate protein phosphotransferase (PtsA is an intracellular protein of the monosaccharide phosphotransferase systems. Biochemical and immunostaining methods were applied to show that PtsA also localizes to the bacterial cell-wall. Thus, it was suspected that PtsA has functions other than its main cytoplasmic enzymatic role. Indeed, recombinant PtsA and anti-rPtsA antiserum were shown to inhibit adhesion of S. pneumoniae to cultured human lung adenocarcinoma A549 cells. Screening of a combinatorial peptide library expressed in a filamentous phage with rPtsA identified epitopes that were capable of inhibiting S. pneumoniae adhesion to A549 cells. The insert peptides in the phages were sequenced, and homologous sequences were found in human BMPER, multimerin1, protocadherin19, integrinβ4, epsin1 and collagen type VIIα1 proteins, all of which can be found in A549 cells except the latter. Six peptides, synthesized according to the homologous sequences in the human proteins, specifically bound rPtsA in the micromolar range and significantly inhibited pneumococcal adhesion in vitro to lung- and tracheal-derived cell lines. In addition, the tested peptides inhibited lung colonization after intranasal inoculation of mice with S. pneumoniae. Immunization with rPtsA protected the mice against a sublethal intranasal and a lethal intravenous pneumococcal challenge. In addition, mouse anti rPtsA antiserum reduced bacterial virulence in the intravenous inoculation mouse model. These findings showed that the surface-localized PtsA functions as an adhesin, PtsA binding peptides derived from its putative target molecules can be considered for future development of therapeutics, and rPtsA should be regarded as a candidate for vaccine development.

  10. Streptococcus pneumoniae Cell-Wall-Localized Phosphoenolpyruvate Protein Phosphotransferase Can Function as an Adhesin: Identification of Its Host Target Molecules and Evaluation of Its Potential as a Vaccine.

    Science.gov (United States)

    Mizrachi Nebenzahl, Yaffa; Blau, Karin; Kushnir, Tatyana; Shagan, Marilou; Portnoi, Maxim; Cohen, Aviad; Azriel, Shalhevet; Malka, Itai; Adawi, Asad; Kafka, Daniel; Dotan, Shahar; Guterman, Gali; Troib, Shany; Fishilevich, Tali; Gershoni, Jonathan M; Braiman, Alex; Mitchell, Andrea M; Mitchell, Timothy J; Porat, Nurith; Goliand, Inna; Chalifa Caspi, Vered; Swiatlo, Edwin; Tal, Michael; Ellis, Ronald; Elia, Natalie; Dagan, Ron

    2016-01-01

    In Streptococcus pneumonia, phosphoenolpyruvate protein phosphotransferase (PtsA) is an intracellular protein of the monosaccharide phosphotransferase systems. Biochemical and immunostaining methods were applied to show that PtsA also localizes to the bacterial cell-wall. Thus, it was suspected that PtsA has functions other than its main cytoplasmic enzymatic role. Indeed, recombinant PtsA and anti-rPtsA antiserum were shown to inhibit adhesion of S. pneumoniae to cultured human lung adenocarcinoma A549 cells. Screening of a combinatorial peptide library expressed in a filamentous phage with rPtsA identified epitopes that were capable of inhibiting S. pneumoniae adhesion to A549 cells. The insert peptides in the phages were sequenced, and homologous sequences were found in human BMPER, multimerin1, protocadherin19, integrinβ4, epsin1 and collagen type VIIα1 proteins, all of which can be found in A549 cells except the latter. Six peptides, synthesized according to the homologous sequences in the human proteins, specifically bound rPtsA in the micromolar range and significantly inhibited pneumococcal adhesion in vitro to lung- and tracheal-derived cell lines. In addition, the tested peptides inhibited lung colonization after intranasal inoculation of mice with S. pneumoniae. Immunization with rPtsA protected the mice against a sublethal intranasal and a lethal intravenous pneumococcal challenge. In addition, mouse anti rPtsA antiserum reduced bacterial virulence in the intravenous inoculation mouse model. These findings showed that the surface-localized PtsA functions as an adhesin, PtsA binding peptides derived from its putative target molecules can be considered for future development of therapeutics, and rPtsA should be regarded as a candidate for vaccine development.

  11. 鹦鹉热嗜衣原体CPSIT_0531融合蛋白原核表达载体的构建、表达及免疫原性检测%Construction and expression of the prokaryotic clone of recombinant protein CPSIT_ 0531 of Chlamydophila psittaci and detection of its immunongenicity

    Institute of Scientific and Technical Information of China (English)

    贺庆芝; 曾怀才; 陆春雪; 王川; 吴移谋

    2014-01-01

    目的 构建鹦鹉热嗜衣原体(Chlamydophila psittaci,Cps) CPSIT 0531原核表达载体,表达并纯化该融合蛋白,检测其免疫原性. 方法 采用PCR技术从Cps 6BC基因组中扩增CPSIT_ 0531基因,并构建pET30a-CPSIT_ 0531原核表达载体,然后转化到宿主菌E.coli BL21中,IPTG诱导His-CPSIT_ 0531融合蛋白表达,免疫BALB/c小鼠制备多克隆抗体,ELISA分析His-CPSIT_ 0531蛋白的免疫原性. 结果 成功构建了pET30a-CPSIT_ 0531原核表达载体,并表达和纯化较稳定的重组蛋白;GST-CPSIT_ 0531免疫小鼠后,ELISA检测免疫小鼠的血清特异性抗体效价为1:640 000. 结论 成功克隆表达了His-CPSIT_ 0531,该蛋白具有较好的免疫原性.

  12. High protein binding and cidal activity against penicillin-resistant S. pneumoniae: a cefditoren in vitro pharmacodynamic simulation.

    Directory of Open Access Journals (Sweden)

    David Sevillano

    Full Text Available BACKGROUND: Although protein binding is a reversible phenomenon, it is assumed that antibacterial activity is exclusively exerted by the free (unbound fraction of antibiotics. METHODOLOGY/PRINCIPAL FINDINGS: Activity of cefditoren, a highly protein bound 3(rd generation cephalosporin, over 24h after an oral 400 mg cefditoren-pivoxil bid regimen was studied against six S. pneumoniae strains (penicillin/cefditoren MICs; microg/ml: S1 (0.12/0.25, S2 (0.25/0.25, S3 and S4 (0.5/0.5, S5 (1/0.5 and S6 (4/0.5. A computerized pharmacodynamic simulation with media consisting in 75% human serum and 25% broth (mean albumin concentrations = 4.85+/-0.12 g/dL was performed. Protein binding was measured. The cumulative percentage of a 24h-period that drug concentrations exceeded the MIC for total (T > MIC and unbound concentrations (fT > MIC, expressed as percentage of the dosing interval, were determined. Protein binding was 87.1%. Bactericidal activity (> or = 99.9% initial inocula reduction was obtained against strains S1 and S2 at 24h (T > MIC = 77.6%, fT > MIC = 23.7%. With T > MIC of 61.6% (fT > MIC = 1.7%, reductions against S3 and S4 ranged from 90% to 97% at 12h and 24h; against S5, reduction was 45.1% at 12h and up to 85.0% at 24h; and against S6, reduction was 91.8% at 12h, but due to regrowth of 52.9% at 24h. Cefditoren physiological concentrations exerted antibacterial activity against strains exhibiting MICs of 0.25 and 0.5 microg/ml under protein binding conditions similar to those in humans. CONCLUSIONS/SIGNIFICANCE: The results of this study suggest that, from the pharmacodynamic perspective, the presence of physiological albumin concentrations may not preclude antipneumococcal activity of highly bound cephalosporins as cefditoren.

  13. Influence of cAMP receptor protein (CRP) on bacterial virulence and transcriptional regulation of allS by CRP in Klebsiella pneumoniae.

    Science.gov (United States)

    Xue, Jian; Tan, Bin; Yang, Shiya; Luo, Mei; Xia, Huiming; Zhang, Xian; Zhou, Xipeng; Yang, Xianxian; Yang, Ruifu; Li, Yingli; Qiu, Jingfu

    2016-11-15

    cAMP receptor protein (CRP) is one of the most important transcriptional regulators, which can regulate large quantities of operons in different bacteria. The gene allS was well-known as allantoin-utilizing capability and involving in bacterial virulence in Klebsiella pneumoniae (K. pneumoniae). The specific DNA recognition motif of transcription regulator CRP was found in allS promoter region. Therefore, this study is aimed to investigate the function of CRP on virulence and its transcriptional regulation mechanism to gene allS in K. pneumoniae. The wild-type (WT) K. pneumoniae NTUH-2044, crp knockout (Kp-Δcrp) and the complemented knockout (KpC-Δcrp) strains were used to determine the function of crp gene. The lacZ fusion, qRT-PCR, electrophoretic mobility shift and DNase I footprinting assays were performed to study the transcriptional regulation of CRP on allS. The result showed a decreased virulence in crp knockout strain. Complement through supplementing crp fragment in expression plasmid partially restore virulence of knockout bacteria. The CRP could bind to the allS promoter-proximal region and the binding site was further refined to be located from 60bp to 94bp upstream of the allS promoter. Based on these results, we proposed that CRP is an essential virulence regulator and knock out of crp gene will result in reduced virulence in K. pneumoniae. In the meantime, the transcription of gene allS is positively regulated by CRP via directly binding to upstream of allS promoter.

  14. Prognostic value of serum procalcitonin and C-reactive protein levels in critically ill patients who developed ventilator-associated pneumonia

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    Hakan Tanriverdi

    2015-01-01

    Full Text Available INTRODUCTION: Ventilator-associated pneumonia (VAP is an important cause of mortality and morbidity in critically ill patients. We sought to determine the prognostic value of procalcitonin (PCT and C-reactive protein (CRP kinetics in critically ill patients who developed VAP. METHODS: Patients who were admitted to the intensive care unit (ICU and developed VAP were eligible. Patients were followed for 28 days after the pneumonia diagnosis and blood samples for PCT and CRP were collected on the day of the pneumonia diagnosis (D0, and days 3 (D3 and 7 (D7 after the diagnosis. Patients were grouped as survivors and non-survivors, and the mean PCT and CRP values and their kinetics were assessed. RESULTS: In total, 45 patients were enrolled. Of them, 22 (48.8% died before day 28 after the pneumonia diagnosis. There was no significant difference between the survivor and non-survivor groups in terms of PCT on the day of pneumonia diagnosis or CRP levels at any point. However, the PCT levels days 3 and 7 were significantly higher in the non-survivor group than the survivor group. Whereas PCT levels decreased significantly from D0 to D7 in the survivor group, CRP did not. A PCT level above 1 ng/mL on day 3 was the strongest predictor of mortality, with an odds ratio of 22.6. CONCLUSION: Serum PCT was found to be a superior prognostic marker compared to CRP in terms of predicting mortality in critically ill patients who developed VAP. The PCT level on D3 was the strongest predictor of mortality in VAP.

  15. A low-affinity penicillin-binding protein 2x variant is required for heteroresistance in Streptococcus pneumoniae.

    Science.gov (United States)

    Engel, Hansjürg; Mika, Moana; Denapaite, Dalia; Hakenbeck, Regine; Mühlemann, Kathrin; Heller, Manfred; Hathaway, Lucy J; Hilty, Markus

    2014-07-01

    Heteroresistance to penicillin in Streptococcus pneumoniae is the ability of subpopulations to grow at a higher antibiotic concentration than expected from the MIC. This may render conventional resistance testing unreliable and lead to therapeutic failure. We investigated the role of the primary β-lactam resistance determinants, penicillin-binding protein 2b (PBP2b) and PBP2x, and the secondary resistance determinant PBP1a in heteroresistance to penicillin. Transformants containing PBP genes from the heteroresistant strain Spain(23F) 2349 in the nonheteroresistant strain R6 background were tested for heteroresistance by population analysis profiling (PAP). We found that pbp2x, but not pbp2b or pbp1a alone, conferred heteroresistance to R6. However, a change of pbp2x expression was not observed, and therefore, expression does not correlate with an increased proportion of resistant subpopulations. In addition, the influence of the CiaRH system, mediating PBP-independent β-lactam resistance, was assessed by PAP on ciaR disruption mutants but revealed no heteroresistant phenotype. We also showed that the highly resistant subpopulations (HOM*) of transformants containing low-affinity pbp2x undergo an increase in resistance upon selection on penicillin plates that partially reverts after passaging on selection-free medium. Shotgun proteomic analysis showed an upregulation of phosphate ABC transporter subunit proteins encoded by pstS, phoU, pstB, and pstC in these highly resistant subpopulations. In conclusion, the presence of low-affinity pbp2x enables certain pneumococcal colonies to survive in the presence of β-lactams. Upregulation of phosphate ABC transporter genes may represent a reversible adaptation to antibiotic stress.

  16. Preparation and testing of a Vi conjugate vaccine using pneumococcal surface protein A (PspA) from Streptococcus pneumoniae as the carrier protein.

    Science.gov (United States)

    Kothari, Neha; Genschmer, Kristopher R; Kothari, Sudeep; Kim, Jeong Ah; Briles, David E; Rhee, Dong Kwon; Carbis, Rodney

    2014-09-29

    In the current study pneumococcal surface protein A (PspA) was conjugated to Vi capsular polysaccharide from Salmonella Typhi to make available a vaccine against typhoid fever that has the potential to also provide broad protection from Streptococcus pneumoniae. High yielding production processes were developed for the purification of PspAs from families 1 and 2. The purified PspAs were conjugated to Vi with high recovery of both Vi and PspA. The processes developed especially for PspA family 2 could readily be adapted for large scale production under cGMP conditions. Previously we have shown that conjugation of diphtheria toxoid (DT) to Vi polysaccharide improves the immune response to Vi but can also enhance the response to DT. In this study it was shown that conjugation of PspA to Vi enhanced the anti-PspA response and that PspA was a suitable carrier protein as demonstrated by the characteristics of a T-cell dependent response to the Vi. We propose that a bivalent vaccine consisting of PspA from families 1 and 2 bound to Vi polysaccharide would protect against typhoid fever and has the potential to also protect against pneumococcal disease and should be considered for use in developing countries.

  17. Hydrocarbon pneumonia

    Science.gov (United States)

    Pneumonia - hydrocarbon ... Coughing Fever Shortness of breath Smell of a hydrocarbon product on the breath Stupor (decreased level of ... Most children who drink or inhale hydrocarbon products and develop ... hydrocarbons may lead to rapid respiratory failure and death.

  18. Occurrence of antibodies against Chlamydophila abortus in sheep and goats in the Slovak Republic.

    Science.gov (United States)

    Cisláková, Lýdia; Halánová, Monika; Kovácová, Daniela; Stefancíková, Astéria

    2007-01-01

    Chlamydophila abortus is one of the most important causative agents of enzootic abortion and other chlamydial infections of sheep and goats. The presence of specific serum antibodies to Chlamydophila abortus was studied in sheep and goats breeding in the Slovak Republic by the complement fixation test. 22,040 sheep and goats were examined during 5 years. Specific anti-Ch. abortus antibodies were found in 2,360 out of 20,878 sheep sera examined (11.7%), and in 85 out of 1,162 examined goats (7.7%). The occurrence of antichlamydial antibodies indicates the importance of performing screening examinations in commercial breeding with the aim of reducing the spread of this disease between animals, and also interrupting the spread and transmission from animals to human.

  19. Occurrence of Chlamydophila felis, feline herpesvirus 1 and calcivirus in domestic cats of Iran

    OpenAIRE

    Nadi Maazi; Shahram Jamshidi; Payman Kayhani; Hassan Momtaz

    2016-01-01

    Background and Objectives: Feline herpesvirus-1, feline calicivirus and Chlamydophila felis are the main causes of feline upper respiratory tract disease. This study was conducted to identify of FeHV-1, FCV and C. felis infections in domestic cat population and also to estimate the prevalence of each specific infection in Iran.Materials and Methods: The ocular conjunctiva and oropharyngeal specimens obtained from 80 cats were examined using PCR and reverse transcription PCR.Results: FeHV-1 wa...

  20. Chlamydophila pecorum in fetuses of mediterranean buffalo (bubalus bubalis bred in Italy

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    G. Galiero

    2010-02-01

    Full Text Available In order to study the role played by the different species of Chlamydophila in causing abortions in Mediterranean buffalo, the Authors examined 164 fetuses from 80 different buffalo herds in Southern Italy. Three fetuses, came from two different herds, were positive. Our study confirms the pathogenic role of C. pecorum in buffalo, not only as a cause of neuropathology in calves but as an infectious abortive agent.

  1. Chlamydophila spp. infection in horses with recurrent airway obstruction: similarities to human chronic obstructive disease

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    Hotzel Helmut

    2008-01-01

    Full Text Available Abstract Background Recurrent airway obstruction (RAO in horses is a naturally occurring dust-induced disease mainly characterized by bronchiolitis which shows histological and pathophysiological similarities to human chronic obstructive pulmonary disease (COPD. In human COPD previous investigations indicated an association with Chlamydophila psittaci infection. The present study was designed (1 to clarify a possible role of this infectious agent in RAO and (2 to investigate the suitability of this equine disorder as a model for human COPD. Methods Clinico-pathological parameters of a total of 45 horses (25 horses with clinical signs of RAO and 20 clinically healthy controls were compared to histological findings in lung tissue samples and infection by Chlamydiaceae using light microscopy, immunohistochemistry, and PCR. Results Horses with clinical signs of RAO vs. controls revealed more inflammatory changes in histology (p = 0.01, and a higher detection rate of Chlamydia psittaci antigens in all cells (p OmpA sequencing identified Chlamydophila psittaci (n = 9 and Chlamydophila abortus (n = 13 in both groups with no significant differences. Within the group of clinically healthy horses subgroups with no changes (n = 15 and slight inflammation of the small airways (n = 5 were identified. Also in the group of animals with RAO subgroups with slight (n = 16 and severe (n = 9 bronchiolitis could be formed. These four subgroups can be separated in parts by the number of cells positive for Chlamydia psittaci antigens. Conclusion Chlamydophila psittaci or abortus were present in the lung of both clinically healthy horses and those with RAO. Immunohistochemistry revealed acute chlamydial infections with inflammation in RAO horses, whereas in clinically healthy animals mostly persistent chlamydial infection and no inflammatory reactions were seen. Stable dust as the known fundamental abiotic factor in RAO is comparable to smoking in human disease. These

  2. New insights into histidine triad proteins: solution structure of a Streptococcus pneumoniae PhtD domain and zinc transfer to AdcAII.

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    Beate Bersch

    Full Text Available Zinc (Zn(2+ homeostasis is critical for pathogen host colonization and invasion. Polyhistidine triad (Pht proteins, located at the surface of various streptococci, have been proposed to be involved in Zn(2+ homeostasis. The phtD gene, coding for a Zn(2+-binding protein, is organized in an operon with adcAII coding for the extracellular part of a Zn(2+ transporter. In the present work, we investigate the relationship between PhtD and AdcAII using biochemical and structural biology approaches. Immuno-precipitation experiments on purified membranes of Streptococcus pneumoniae (S. pneumoniae demonstrate that native PhtD and AdcAII interact in vivo confirming our previous in vitro observations. NMR was used to demonstrate Zn(2+ transfer from the Zn(2+-bound form of a 137 amino acid N-terminal domain of PhtD (t-PhtD to AdcAII. The high resolution NMR structure of t-PhtD shows that Zn(2+ is bound in a tetrahedral site by histidines 83, 86, and 88 as well as by glutamate 63. Comparison of the NMR parameters measured for apo- and Zn(2+-t-PhtD shows that the loss of Zn(2+ leads to a diminished helical propensity at the C-terminus and increases the local dynamics and overall molecular volume. Structural comparison with the crystal structure of a 55-long fragment of PhtA suggests that Pht proteins are built from short repetitive units formed by three β-strands containing the conserved HxxHxH motif. Taken together, these results support a role for S. pneumoniae PhtD as a Zn(2+ scavenger for later release to the surface transporter AdcAII, leading to Zn(2+ uptake.

  3. Identificación de Chlamydophila abortus en un aborto ovino en Almoloya de Juárez, México

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    Edgardo Soriano-Vargas

    2011-11-01

    Full Text Available ResumenSe identificó la presencia de Chlamydophila abortus en un aborto ovino procedente de Almoloya de Juárez, México.SummaryChlamydophila abortus was identified in an ovine aborted fetus from Almoloya de Juárez, México

  4. Detection of Chlamydophila psittaci from pigeons by polymerase chain reaction in Ahvaz

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    Masoud Ghorbanpoor

    2015-10-01

    Full Text Available Background and Objective: Chlamydophila psittaci is a lethal bacterium that causes endemic avian chlamydiosis, and respiratory psittacosis. Laboratory diagnosis of Chlamydophila psittaci is difficult by culture. This study was design to investigate the presence of Chlamydophila psittaci in collected pharyngeal swabs from asyptomatic pigeons by PCR.Materials and Methods: Pharyngeal samples from pigeons with no symptoms of disease (n=280 were collected during hot and cold seasons in different parts of Ahvaz. DNA was extracted from specimens and subjected to PCR targeting pmp genes and 16s-23s rRNA intergenic spacer of Cp. psittaci and chlamydiales specific primers.Results: Of 280 samples 2 (0.7% harbor were positive for chlamydiales (16s-23s intergenic spacer and Cp. psittaci specific genes (pmp gene.Conclusions: In this research the pigeons were asymptomatic carriers for Cp. psittaci in their respiratory discharges. These results suggest that Cp. psittaci infection of human can occur in very close and continuous contact with pigeons.

  5. An outbreak of Chlamydophila psittaci in an outdoor colony of Magellanic penguins (Spheniscus magellanicus).

    Science.gov (United States)

    Jencek, Jacqueline E; Beaufrère, Hugues; Tully, Thomas N; Garner, Michael M; Dunker, Freeland H; Baszler, Timothy V

    2012-12-01

    An outbreak of Chlamydophila psittaci occurred in an outdoor colony of 63 Magellanic penguins (Spheniscus magellanicus) at the San Francisco Zoo. Affected penguins presented with inappetence, lethargy, and light green urates. Hematologic and serum biochemical findings were consistent with chronic inflammation. Penguins did not respond to initial supportive and antimicrobial therapy, and 3 died. Necropsy results of the 3 birds revealed hepatomegaly and splenomegaly, and histologic lesions included necrotizing hepatitis, splenitis, and vasculitis. Chlamydophila psittaci infection was confirmed by results of Gimenez staining, immunohistochemistry, and tissue polymerase chain reaction assay. As additional birds continued to present with similar clinical signs, the entire colony of penguins was prophylactically treated with a 30-day minimum course of doxycycline, administered orally or intramuscularly or as a combination of both. Despite treatment, 9 additional penguins died during a 3-month period. Pathologic results from these birds revealed renal and visceral gout (n = 4), cardiac insufficiency (n = 2), sepsis from a suspected esophageal perforation (n = 2), and no gross lesions (n = 1). During the outbreak, 4 birds presented with seizures, 5 developed dermatitis, and nearly 90% of birds in the colony showed severe keratoconjunctivitis, believed to be related to drug therapy with doxycycline. We report the clinical and pathologic features of Chlamydophila psittaci infection in an outdoor colony of penguins and the associated challenges of treatment.

  6. Detection and characterization ofChlamydophila psittaci in asymptomatic feral pigeons (Columba livia domestica) in central Thailand

    Institute of Scientific and Technical Information of China (English)

    Ladawan Sariya; Phirom Prompiram; Siriporn Tangsudjai; Kanaporn Poltep; Tatiyanuch Chamsai; Chalisa Mongkolphan; Kamolphan Rattanavibul; Verachai Sakdajivachareon

    2015-01-01

    Objective:To detect and characterizeChlamydophila psittaci(C. psittaci) in asymptomatic feral pigeons in centralThailand.Methods:A total814 swabs from the trachea and cloacae of407 non-clinical feral pigeons in centralThailand were collected and tested for the presence ofC. psittaci.Results:A10.8% of feral pigeons in the sample group were positive as determined by nestedPCR primer specific toC. psittaci.The outer membrane proteinA(ompA) gene of positive samples exhibited amino acid identity ofC. psittaci ranging from71 to100% and were grouped in genotypeB.Exceptionally,BF1676-56 isolate was closely related toChlamydia avium with 99% identification of the16S ribosomal(r)RNA gene.Conclusions:This is the first report onC. psittaci isolated from asymptomatic feral pigeons inThailand, which provides knowledge for the disease status in pigeon populations inThailand.

  7. C-反应蛋白检测在小儿细菌性肺炎及支原体肺炎中的应用%Detection of C-reactive protein applied in diagnosis of children with bacterial pneumonia and mycoplasma pneumonia

    Institute of Scientific and Technical Information of China (English)

    贺箭飞

    2012-01-01

    目的 对比分析C反应蛋白检测在小儿细菌性肺炎和支原体肺炎的诊断价值.方法 选取148例肺炎患儿及同期进行健康体检的150例健康小儿作为研究对象,对所有受试者进行C-反应蛋白、肺炎支原体、血常规检测,对148例肺炎患儿治疗前后进行C-反应蛋白水平检测,比较健康小儿、支原体肺炎患儿、细菌性肺炎患儿的C-反应蛋白水平,比较细菌性肺炎患儿与支原体肺炎患儿治疗前后的C-反应蛋白水平,比较C-反应蛋白对不同类型肺炎患儿的阳性检测率,综合评价C-反应蛋白在肺炎患儿诊断鉴别中的价值.结果 健康患儿、肺炎支原体患儿、细菌性肺炎患儿的C反应蛋白分别为(3.58±0.79)、(14.82±3.69)、(68.54±28.31)mg/L;组间比较,肺炎组患儿明显高于健康组患儿(P<0.05),而细菌性肺炎组患儿明显高于支原体肺炎组患儿(P<0.05);细菌性肺炎患儿C-反应蛋白的阳性率为93.3%、支原体肺炎患儿为45.6%,患儿经治疗后,C-反应蛋白均明显下降,上述指标组间比较及治疗前后比较,差异均有统计学意义(P<0.05).结论 C-反应蛋白检测可作为鉴别细菌性肺炎和支原体肺炎的有效手段,有助于临床医师采取相应的治疗方法及早治疗患儿,以免耽误病情.%OBJECTIVE To compare and analyze the C-reactive protein detection for the diagnosis of the children with bacterial pneumonia and mycoplasma pneumonia. METHODS A total of 148 children with pneumonia and 150 healthy children were chosen as the research subjects. All subjects were taken with the C-reactive protein, Mycoplasma pneumoniae. and blood testing. The levels of C-reactive protein of 148 cases of pneumonia children were tested before and after the treatment. The C-reactive protein levels of the healthy children, the children with mycoplasma pneumonia, and the children with bacterial pneumonia were compared. The C-reactive protein levels of the children

  8. Prevent Pneumonia

    Centers for Disease Control (CDC) Podcasts

    2015-08-06

    CDC’s Matthew Westercamp explains what pneumonia is, its symptoms, and how to prevent it.  Created: 8/6/2015 by National Center for Immunization and Respiratory Diseases (NCIRD), Division of Bacterial Diseases (DBD), Respiratory Diseases Branch (RDB).   Date Released: 8/6/2015.

  9. Chlamydia Pneumoniae Pneumonia: An Evolving Clinical Spectrum

    Directory of Open Access Journals (Sweden)

    David Megran

    1995-01-01

    Full Text Available Chlamydia pneumoniae is a recently recognized respiratory tract pathogen. It accounts for 6 to 10% of all cases of community acquired pneumonia requiring admission to hospital. Two patients hospitalized with C pneumoniae pneumonia are presented to illustrate its range of severity and the extrapulmonary manifestations.

  10. Chlamydia pneumoniae pneumonia: An evolving clinical spectrum

    Science.gov (United States)

    Megran, David; Peeling, Rosanna W; Marrie, Thomas J

    1995-01-01

    Chlamydia pneumoniae is a recently recognized respiratory tract pathogen. It accounts for 6 to 10% of all cases of community acquired pneumonia requiring admission to hospital. Two patients hospitalized with C pneumoniae pneumonia are presented to illustrate its range of severity and the extrapulmonary manifestations. PMID:22514396

  11. C-反应蛋白检测在小儿细菌性肺炎与支原体肺炎中的临床比较%Clinical value of detection of C-reaction protein in diagnosis of pediatric bacterial pneumonia and Mycoplasma pneumonia:a comparative study

    Institute of Scientific and Technical Information of China (English)

    石丰月

    2013-01-01

    OBJECTIVE To study the clinical value of C-reactie protein (CRP) detection in diagnosis of the pediatric bacterial pneumonia and the Mycoplasma pneumonia so as to guide the diagnosis of infantile pneumonia.METHODS The children with bacterial pneumonia (the bacterial pneumonia group) and the children with Mycoplasma pneumonia (the Mycoplasma pneumonia group),who were treated in the pediatrics department from Sep 2010 to Sep 2012,were enrolled in the study,the healthy children receiving the medical examination (the control group)were also selected,with 80 cases in each group,then the change of CRP level was determined.RESULTS The CRP level of the control group was (3.24 ±0.45)mg/L,the bacterial pneumonia group before treatment (44.03±5.83) mg/L,the bacterial pneumonia group 3 days after the treatment (15.12±6.21) mg/L,the Mycoplasma pneumonia group before treatment (13.97±4.96) mg/L,the Mycoplasma pneumonia group 3 days after the treatment (5.29 ± 2.33) mg/L,the CRP level of the bacterial pneumonia group and the Mycoplasma pneumonia group before the treatment was significantly higher than that of the control group (P<0.05),the CRP level of the bacterial pneumonia group was higher than that of the Mycoplasma pneumonia group (P<0.05),the CRP level of the bacterial pneumonia group 3 days after treatment was higher than that of the Mycoplasma pneumonia group (P<0.05),the difference in the CRP level before and after the treatment between the bacterial pneumonia group and the Mycoplasma pneumonia group was statistically significant (P< 0.05).The positive rates of the CRP of the bacterial pneumonia group were 100.00% before the treatment and 52.50% 3 days after the treatment,which were respectively 66.25 % and 21.25 % in the Mycoplasma pneumonia group,the difference in the positive rate before and after the treatment between the bacterial pneumonia group and the Mycoplasmapneumonia group was statistically significant (P<0.05).CONCLUSION The positive

  12. C-reactive protein and anti-Chlamydia pneumoniae antibodies as risk factors of cardiovascular death in incident patients on peritoneal dialysis.

    Science.gov (United States)

    Paniagua, Ramón; Frías, Yolanda; de Ventura, Maria Jesús; Rodríguez, Ernesto; Hurtado, María Elena; Alcántara, Guadalupe; Vázquez, Roberto; Ortiz, Ruth; Salcedo, Mario; Rios, Maria Elena; Kaji, Julio; Amato, Dante

    2003-01-01

    Recently it has been pointed out that inflammation and infections caused by germs such as Chlamydia pneumoniae are independent cardiovascular risk factors for the general population, but information about these relationships in dialysis patients is scarce. This work was done to analyze the association of C-reactive protein (CRP) and IgG anti-Chlamydia pneumoniae antibodies (anti-Chlp-IgG) as independent cardiovascular risk factors in incident patients on continuous ambulatory peritoneal dialysis (CAPD). Single-cohort, prospective observational study. Three CAPD centers from the Instituto Mexicano del Seguro Social, and one from the Instituto de Seguridad y Servicios Sociales de los Trabajadores del Estado, Mexico. A cohort of 75 adult incident patients on CAPD, without clinical signs of congestive heart failure, coronary heart disease, or peripheral arterial insufficiency. No restrictions for age, gender, or cause of renal failure were applied. Mortality. Demographic variables, body composition by electrical bioimpedance, serum glucose, urea, creatinine, lipids, homocysteine, nutritional markers (albumin, prealbumin, and transferrin), CRP, and anti-Chlp-IgG were measured and registered at the time of the first admission. When a patient died, the cause of death was determined by review of the clinical chart. Mean follow-up time was 10.25 patient-months. There were 14 cardiovascular deaths. CRP was positive (> 10 mg/L) in 64% of the patients, and anti-Chlp-IgG in 64%; 29% of the patients were positive for both markers. The relative risk for cardiovascular mortality was 6.23 for patients positive for either CRP or anti-Chlp-IgG, and increased to 9.52 when both markers were positive. Multivariate analysis revealed that CRP and anti-Chlp-IgG were stronger cardiovascular death predictors than age, diabetes, and nutritional status. These data suggest that inflammation and the presence of Chlamydia pneumoniae infections are important predictors of cardiovascular death in

  13. Real-Time Detection and Identification of Chlamydophila Species in Veterinary Specimens by Using SYBR Green-Based PCR Assays

    DEFF Research Database (Denmark)

    Nordentoft, Steen; Kabell, Susanne; Pedersen, Karl

    2011-01-01

    specificity of different species is not as clear as recently believed. As most members of the genus Chlamydophila have shown to be transmissible from animals to humans, sensitive and fast detection methods are required. In this study, SYBR green-based real-time assays were developed that detect all members...... of Chlamydiaceae and differentiate the most prevalent veterinary Chlamydophila species: Cp. psittaci, Cp. abortus, Cp. felis, and Cp. caviae. By adding bovine serum albumin to the master mixes, target DNA could be detected directly in crude lysates of enzymatically digested conjunctival or pharyngeal swabs...

  14. Higher levels of mucosal antibody to pneumococcal vaccine candidate proteins are associated with reduced acute otitis media caused by Streptococcus pneumoniae in young children.

    Science.gov (United States)

    Xu, Q; Casey, J R; Pichichero, M E

    2015-09-01

    Mucosal immunity has a crucial role in controlling human respiratory tract infections. This study characterizes the naturally acquired mucosal antibody levels to three Streptococcus pneumoniae (Spn) protein antigens, pneumococcal histidine triad protein D (PhtD), pneumococcal choline binding protein A (PcpA), and pneumolysin (Ply), and assesses the association of the mucosal antibody levels with occurrence of acute otitis media (AOM) caused by Spn. Both nasopharyngeal (NP) immunoglobulin G (IgG) and IgA levels to all three proteins slightly decreased in children from 6 to 9 months of age and then gradually increased through 24 months of age. Spn NP colonization was associated with higher mucosal antibody levels to all three proteins. However, children with Spn AOM had 5-8-fold lower IgG and 3-6-fold lower IgA levels to the three proteins than children without AOM but asymptomatically colonized with Spn. Antigen-specific antibody levels in the middle ear fluid (MEF) were correlated with antibody levels in the NP. Children with AOM caused by Spn had lower antibody levels in both the MEF and NP than children with AOM caused by other pathogens. These results indicate that higher naturally acquired mucosal antibody levels to PhtD, PcpA and Ply are associated with reduced AOM caused by Spn.

  15. Respiratory Syncytial Virus Increases the Virulence of Streptococcus pneumoniae by Binding to Penicillin Binding Protein 1a A New Paradigm in Respiratory Infection

    NARCIS (Netherlands)

    Smith, Claire M.; Sandrini, Sara; Datta, Sumit; Freestone, Primrose; Shafeeq, Sulman; Radhakrishnan, Priya; Williams, Gwyneth; Glenn, Sarah M.; Kuipers, Oscar P.; Hirst, Robert A.; Easton, Andrew J.; Andrew, Peter W.; O'Callaghan, Christopher

    2014-01-01

    Rationale: Respiratory syncytial virus (RSV) and Streptococcus pneumoniae are major respiratory pathogens. Coinfection with RSV and S. pneumoniae is associated with severe and often fatal pneumonia but the molecular basis for this remains unclear. ObjeOtives: Todetermine if interaction between RSV a

  16. Mucosal immunization with PsaA protein, using chitosan as a delivery system, increases protection against acute otitis media and invasive infection by Streptococcus pneumoniae.

    Science.gov (United States)

    Xu, J-H; Dai, W-J; Chen, B; Fan, X-Y

    2015-03-01

    As infection with Streptococcus pneumoniae (mainly via the mucosal route) is a leading cause of acute otitis media, sinus and bacterial pneumonia, the mucosal immunity plays an important role in the prevention of pneumococcal diseases. Therefore, intranasal vaccination may be an effective immunization strategy, but requires appropriate mucosal vaccine delivery systems. In this work, chitosan was used as a mucosal delivery system to form chitosan-PsaA nanoparticles based on ionotropic gelation methods and used to immunize BALB/c mice intranasally. Compared to mice immunized with naked PsaA, levels of IFN-γ, IL-17A and IL-4 in spleen lymphocytes, the systemic (IgG in serum) and mucosal (IgA in mucosal lavage) specific antibodies were enhanced significantly in mice inoculated with chitosan-PsaA. Furthermore, increased protection against acute otitis media following middle ear challenge with pneumococcus serotype 14, and improved survival following intraperitoneal challenge with pneumococcus serotype 3 or serotype 14, was found in the mice immunized with chitosan-PsaA nanoparticles. Thus, intranasal immunization with chitosan-PsaA can successfully induce mucosal and systemic immune responses and increase protection against pneumococcal acute otitis media and invasive infections. Hence, intranasal immunization with PsaA protein, based on chitosan as a delivery system, is an efficient immunization strategy for preventing pneumococcal infections.

  17. Genomics of KPC-producing Klebsiella pneumoniae sequence type 512 clone highlights the role of RamR and ribosomal S10 protein mutations in conferring tigecycline resistance.

    Science.gov (United States)

    Villa, Laura; Feudi, Claudia; Fortini, Daniela; García-Fernández, Aurora; Carattoli, Alessandra

    2014-01-01

    Full genome sequences were determined for five Klebsiella pneumoniae strains belonging to the sequence type 512 (ST512) clone, producing KPC-3. Three strains were resistant to tigecycline, one showed an intermediate phenotype, and one was susceptible. Comparative analysis performed using the genome of the susceptible strain as a reference sequence identified genetic differences possibly associated with resistance to tigecycline. Results demonstrated that mutations in the ramR gene occurred in two of the three sequenced strains. Mutations in RamR were previously demonstrated to cause overexpression of the AcrAB-TolC efflux system and were implicated in tigecycline resistance in K. pneumoniae. The third strain showed a mutation located at the vertex of a very well conserved loop in the S10 ribosomal protein, which is located in close proximity to the tigecycline target site in the 30S ribosomal subunit. This mutation was previously shown to be associated with tetracycline resistance in Neisseria gonorrhoeae. A PCR-based approach was devised to amplify the potential resistance mechanisms identified by genomics and applied to two additional ST512 strains showing resistance to tigecycline, allowing us to identify mutations in the ramR gene.

  18. Pneumocystis Pneumonia (For Parents)

    Science.gov (United States)

    ... Feeding Your 1- to 2-Year-Old Pneumocystis Pneumonia KidsHealth > For Parents > Pneumocystis Pneumonia A A A What's in this article? About PCP Diagnosing PCP Treating PCP Pneumocystis pneumonia (PCP) is an infection caused by Pneumocystis jiroveci , ...

  19. How Is Pneumonia Treated?

    Science.gov (United States)

    ... to cure the infection and prevent complications. Bacterial pneumonia Bacterial pneumonia is treated with medicines called antibiotics. ... fewer symptoms such as cough and fever. Viral pneumonia Antibiotics don't work when the cause of ...

  20. What Is Pneumonia?

    Science.gov (United States)

    ... Share this page from the NHLBI on Twitter. Pneumonia Pneumonia is a bacterial, viral, or fungal infection of ... and trouble breathing. Many factors affect how serious pneumonia is, such as the type of germ causing ...

  1. Pneumonia in Immunocompromised People

    Science.gov (United States)

    ... Alternative preventive drug treatments are dapsone , atovaquone , and pentamidine (which can be taken as an aerosol, inhaled ... ACZONE trimethoprim No US brand name atovaquone MEPRON pentamidine NEBUPENT Pneumonia Overview of Pneumonia Aspiration Pneumonia and ...

  2. Clinical features of severe or fatal Mycoplasma pneumoniae pneumonia

    Directory of Open Access Journals (Sweden)

    Koichi eIzumikawa

    2016-06-01

    Full Text Available Mycoplasma pneumoniae is one of the most common causes of community-acquired pneumonia in children and young adults. The incidence of fulminant M. pneumoniae pneumonia (MPP is relatively rare despite the high prevalence of M. pneumoniae infection. This literature review highlights the clinical features of fulminant MPP by examining the most recent data in epidemiology, clinical presentation, pathogenesis, and treatment. Fulminant MPP accounts for 0.5-2% of all MPP cases and primarily affects young adults with no underlying disease. Key clinical findings include a cough, fever, and dyspnea along with diffuse abnormal findings in radiological examinations. Levels of inflammatory markers such as white blood cells (WBC and C-reactive protein (CRP are elevated, as well as levels of lactate dehydrogenase (LDH, IL-18, AST, and ALT. The exact pathogenesis of fulminant MPP remains unclear, but theories include a delayed hypersensitivity reaction to Mycoplasma pneumoniae and the contribution of delayed antibiotic administration to disease progression. Treatment options involve pairing the appropriate anti-mycoplasmal agent with a corticosteroid that will downregulate the hypersensitivity response, and mortality rates are quite low in this treatment group. Further research is necessary to determine the exact pathogenesis of severe and fulminant types of MPP.

  3. Feline herpesvirus 1 and Chlamydophila felis prevalence in cats with chronic conjunctivitis.

    Science.gov (United States)

    Wieliczko, A K; Płoneczka-Janeczko, K

    2010-01-01

    The prevalence of Chlamydophila felis and Feline Herpesvirus type 1 was investigated in 30 cats with chronic conjunctivitis, with use of conjunctival swabs and conventional polymerase chain reaction (PCR). In cats with chronic conjunctivitis the DNA of C. felis and FHV-1 was detected in 2 of 30 cats (6.7%) and in 10 of 30 animals (33.3%), respectively. One case of FHV-1 DNA, and none of C. felis was found in control group. There was no case of co-infection with both pathogens.

  4. Potential role of tigecycline in the treatment of community-acquired bacterial pneumonia

    Science.gov (United States)

    Townsend, Mary L; Pound, Melanie W; Drew, Richard H

    2011-01-01

    Tigecycline is a member of the glycylcycline class of antimicrobials, which is structurally similar to the tetracycline class. It demonstrates potent in vitro activity against causative pathogens that are most frequently isolated in patients with community-acquired bacterial pneumonia (CABP), including (but not limited to) Streptococcus pneumoniae (both penicillin-sensitive and -resistant strains), Haemophilus influenzae and Moraxella catarrhalis (including β-lactamase-producing strains), Klebsiella pneumoniae, and ‘atypical organisms’ (namely Chlamydophila pneumoniae, Mycoplasma pneumoniae, and Legionella pneumophila). Comparative randomized clinical trials to date performed in hospitalized patients receiving tigecycline 100 mg intravenous (IV) × 1 and then 50 mg IV twice daily thereafter have demonstrated efficacy and safety comparable to the comparator agent. Major adverse effects were primarily gastrointestinal in nature. Tigecycline represents a parenteral monotherapy option in hospitalized patients with CABP (especially in patients unable to receive respiratory fluoroquinolones). However, alternate and/or additional therapies should be considered in patients with more severe forms of CABP in light of recent data of increased mortality in patients receiving tigecycline for other types of severe infection. PMID:21694911

  5. High seroprevalence of Mycoplasma pneumoniae IgM in acute Q fever by enzyme-linked immunosorbent assay (ELISA.

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    Chung-Hsu Lai

    Full Text Available Q fever is serologically cross-reactive with other intracellular microorganisms. However, studies of the serological status of Mycoplasma pneumoniae and Chlamydophila pneumoniae during Q fever are rare. We conducted a retrospective serological study of M. pneumoniae and C. pneumoniae by enzyme-linked immunosorbent assay (ELISA, a method widely used in clinical practice, in 102 cases of acute Q fever, 39 cases of scrub typhus, and 14 cases of murine typhus. The seropositive (57.8%, 7.7%, and 0%, p<0.001 and seroconversion rates (50.6%, 8.8%, and 0%, p<0.001 of M. pneumoniae IgM, but not M. pneumoniae IgG and C. pneumoniae IgG/IgM, in acute Q fever were significantly higher than in scrub typhus and murine typhus. Another ELISA kit also revealed a high seropositivity (49.5% and seroconversion rate (33.3% of M. pneumoniae IgM in acute Q fever. The temporal and age distributions of patients with positive M. pneumoniae IgM were not typical of M. pneumoniae pneumonia. Comparing acute Q fever patients who were positive for M. pneumoniae IgM (59 cases with those who were negative (43 cases, the demographic characteristics and underlying diseases were not different. In addition, the clinical manifestations associated with atypical pneumonia, including headache (71.2% vs. 81.4%, p=0.255, sore throat (8.5% vs. 16.3%, p=0.351, cough (35.6% vs. 23.3%, p=0.199, and chest x-ray suggesting pneumonia (19.3% vs. 9.5%, p=0.258, were unchanged between the two groups. Clinicians should be aware of the high seroprevalence of M. pneumoniae IgM in acute Q fever, particularly with ELISA kits, which can lead to misdiagnosis, overestimations of the prevalence of M. pneumoniae pneumonia, and underestimations of the true prevalence of Q fever pneumonia.

  6. 肺炎嗜衣原体主要外膜蛋白的克隆表达与抗原性研究%Cloning and expression of gene encoding chlamydophila pneumoniae major outer membrane protein and the antigenicity analysis of the protein

    Institute of Scientific and Technical Information of China (English)

    李岩伟; 宋立华; 左庭婷; 何君; 朱虹; 端青

    2008-01-01

    肺炎嗜衣原体主要外膜蛋白是其特征抗原之一.实验中通过PCR方法从肺炎嗜衣原体基因组中扩增主要外膜蛋白基因,插入pET32a(+)表达载体,转化BL21(DE3)感受态细胞,得到表达56kD融合蛋白的工程菌株.该菌株的表达量可达53%,提纯后的主要外膜蛋白纯度可达90%以上,在Western Blotting试验和胶体金免疫层析试验中显示了良好的抗原性.

  7. Chlamydia pneumoniae, heat shock proteins 60 and risk of secondary cardiovascular events in patients with coronary heart disease under special consideration of diabetes: a prospective study

    Directory of Open Access Journals (Sweden)

    Twardella Dorothee

    2006-04-01

    Full Text Available Abstract Background There have been suggestions of an association between Chlamydia pneumoniae, chlamydial heat shock protein (Ch-hsp 60 and human heat shock protein (h-hsp 60 infection sero-status and development of secondary cardiovascular events. Patients with diabetes might be at higher risk since they are prone to infections. The objective of this study was to investigate prospectively the role of Chlamydia pneumoniae (CP, chlamydial heat shock protein (Ch-hsp 60 and a possible intermediate role of human heat shock protein (h-hsp 60 sero-status in the development of secondary cardiovascular disease (CVD events in patients with coronary heart disease (CHD under special consideration of diabetes mellitus. Methods Patients aged 30–70 undergoing an in-patient rehabilitation program after acute manifestation of coronary heart disease (International Classification of Disease, 9th Rev. pos. 410–414 between January 1999 and May 2000 in one of two participating rehabilitation clinics in Germany were included in this analysis. Chlamydia pneumoniae (CP, chlamydial heat shock protein (Ch-hsp 60 and human heat shock protein (h-hsp 60 status at baseline were measured by serum immunoglobulin G and A antibodies. Secondary CVD events (myocardial infarction, stroke, and cardiovascular death were recorded during a mean follow-up period of 33.5 months (response = 87%. Results Among the 1052 subjects 37.4% and 39.3% were sero-positive to CP IgA and IgG respectively, 22.2% were sero-positive to Ch-hsp 60 IgG and 8.4% were positive to h-hsp 60 IgG at baseline. During follow-up, secondary CVD events occurred among 71 (6.8% participants. Occurrence of a secondary CVD event was more common among CP (IgA and CP (IgG sero-positive than among sero-negative patients (p-values 0.04 and 0.1, respectively. The risk of secondary CVD events was increased among patients with both a positive CP sero-status and diabetes compared to infection negative, non-diabetic patients

  8. Penicillin-binding protein 2x of Streptococcus pneumoniae: the mutation Ala707Asp within the C-terminal PASTA2 domain leads to destabilization.

    Science.gov (United States)

    Schweizer, Inga; Peters, Katharina; Stahlmann, Christoph; Hakenbeck, Regine; Denapaite, Dalia

    2014-06-01

    Streptococcus pneumoniae penicillin-binding protein 2x (PBP2x) is an enzyme involved in the last stages of peptidoglycan assembly and essential for bacterial growth and survival. PBP2x localizes to the division site, a process that depends on its Penicillin-Binding Protein And Serine-Threonine-kinase Associated (PASTA) domains, which was previously demonstrated via GFP-PBP2x in living cells. During this study a mutant strain was isolated in which the GFP-PBP2x fusion protein did not localize at division sites and it contained reduced amounts of the full-length GFP-PBP2x. We now show that this defect is due to a point mutation within the C-terminal PASTA2 domain of PBP2x. The mutant protein was analyzed in detail in terms of beta-lactam binding, functionality, and localization in live cells. We demonstrate that the mutation affects the GFP-tagged PBP2x variant severely and renders it susceptible to the protease/chaperone HtrA.

  9. The highly conserved serine threonine kinase StkP of Streptococcus pneumoniae contributes to penicillin susceptibility independently from genes encoding penicillin-binding proteins

    Directory of Open Access Journals (Sweden)

    Dias Ricardo

    2009-06-01

    Full Text Available Abstract Background The serine/threonine kinase StkP of Streptococcus pneumoniae is a major virulence factor in the mouse model of infection. StkP is a modular protein with a N-terminal kinase domain a C-terminal PASTA domain carrying the signature of penicillin-binding protein (PBP and prokaryotic serine threonine kinase. In laboratory cultures, one target of StkP is the phosphoglucosamine mutase GlmM involved in the first steps of peptidoglycan biosynthesis. In order to further elucidate the importance of StkP in S. pneumoniae, its role in resistance to β-lactams has been assessed by mutational analysis in laboratory cultures and its genetic conservation has been investigated in isolates from infected sites (virulent, asymptomatic carriers, susceptible and non-susceptible to β-lactams. Results Deletion replacement mutation in stkP conferred hypersensitivity to penicillin G and was epistatic on mutations in PBP2X, PBP2B and PBP1A from the resistant 9V clinical isolate URA1258. Genetic analysis of 55 clinical isolates identified 11 StkP alleles differing from the reference R6 allele. None relevant mutation in the kinase or the PASTA domains were found to account for susceptibility of the isolates. Rather the minimal inhibitory concentration (MIC values of the strains appeared to be determined by their PBP alleles. Conclusion The results of genetic dissection analysis in lab strain Cp1015 reveal that StkP is involved in the bacterial response to penicillin and is epistatic on mutations PBP 2B, 2X and 1A. However analysis of the clinical isolates did not allow us to find the StkP alleles putatively involved in determining the virulence or the resistance level of a given strain, suggesting a strong conservation of StkP in clinical isolates.

  10. Transcriptional activation of the mrkA promoter of the Klebsiella pneumoniae type 3 fimbrial operon by the c-di-GMP-dependent MrkH protein.

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    Ji Yang

    Full Text Available The Gram-negative bacterial pathogen Klebsiella pneumoniae forms biofilms to facilitate colonization of biotic and abiotic surfaces. The formation of biofilms by K. pneumoniae requires the expression of type 3 fimbriae: elongate proteinaceous filaments extruded by a chaperone-usher system in the bacterial outer membrane. The expression of the mrkABCDF cluster that encodes this fimbrial system is strongly positively regulated by MrkH, a transcriptional activator that responds to the second messenger, c-di-GMP. In this study, we analyzed the mechanism by which the MrkH protein activates transcriptional initiation from the mrkA promoter. A mutational analysis supported by electrophoretic mobility shift assays demonstrated that a 12-bp palindromic sequence (the MrkH box centered at -78.5 is the binding site of MrkH. Deletion of half a turn, but not a full turn, of DNA located between the MrkH box and the mrkA promoter destroyed the ability of MrkH to activate mrkA transcription. In addition, a 10-bp AT-rich sequence (the UP element centered at -63.5 contributed significantly to MrkH-dependent mrkA transcription. In vivo analysis of rpoA mutants showed that the R265 and E273 determinants in the C-terminal domain of RNA polymerase α subunit are needed for MrkH-mediated activation of mrkA transcription. Furthermore, results from mutagenesis of the mrkH gene suggest that the N-terminal region of the protein is involved in transcriptional activation. Taken together, our results suggest that MrkH activates mrkA expression by interacting directly with RNA polymerase, to overcome the inefficient transcriptional initiation caused by the presence of defective core promoter elements.

  11. Validation of a PCR Assay for Chlamydophila abortus rRNA gene detection in a murine model

    Directory of Open Access Journals (Sweden)

    Francielle Gibson da Silva-Zacarias

    2009-11-01

    Full Text Available Chlamydophila abortus (C. abortus is associated with reproductive problems in cattle, sheep, and goats. Diagnosis of C. abortus using embryonated chicken eggs or immortalized cell lines has a very low sensitivity. Polymerase chain reaction (PCR assays have been used to detect C. abortus infection in clinical specimens and organ fragments, such as placenta, fetal organs, vaginal secretions, and semen. The aim of this study was to develop a PCR assay for the amplification of an 856-bp fragment of the rRNA gene of the Chlamydiaceae family. The PCR assay was evaluated using organs from 15 mice experimentally infected with the S26/3 reference strain of C. abortus. The results of the rRNA PCR were compared to the results from another PCR system (Omp2 PCR that has been previously described for the Omp2 (outer major protein gene from the Chlamydiaceae family. From the 15 C. abortus-inoculated mice, 13 (K=0.84, standard error =0.20 tested positive using the rRNA PCR assay and 9 (K=0.55, standard error=0.18 tested positive using the Omp2 PCR assay. The detection limit, measured using inclusion-forming units (IFU, for C. abortus with the rRNA PCR (1.05 IFU was 100-fold lower than for the Omp2 PCR (105 IFU. The higher sensitivity of the rRNA PCR, as compared to the previously described PCR assay, and the specificity of the assay, demonstrated using different pathogenic microorganisms of the bovine reproductive system, suggest that the new PCR assay developed in this study can be used for the molecular diagnosis of C. abortus in abortion and other reproductive failures in bovines, caprines, and ovines.Chlamydophila abortus (C. abortus é frequentemente associada a distúrbios reprodutivos em bovinos, ovinos e caprinos. Para o diagnóstico, os métodos de cultivo em ovo embrionado de galinha e em células de linhagem contínua apresentam baixa sensibilidade. A reação em cadeia da polimerase (PCR tem sido utilizada em placenta, órgãos fetais, secre

  12. C-反应蛋白检测在小儿肺炎中的诊断价值%Value of detection of C-reactive protein in diagnosis of pneumonia in children

    Institute of Scientific and Technical Information of China (English)

    叶素芬

    2012-01-01

    OBJECTIVE To explore the clinical value of the detection of C-reactive protein in the diagnosis of pneumonia in children. METHODS A total of 120 cases of pediatric patients with pneumonia treated hospital from May 2010 to May 2011 were chosen as the research objects, according to the types of pneumonia, they were divided into bacterial pneumonia group, viral pneumonia group, and Mycoplasma pneumonia group; totally 50 healthy children receiving physical examination at the same period were chosen as the control groups the white blood cells and C-reactive protein indicators of three groups of children were determined as they were treated, the indicators mentioned above "were also detected for the control group at the same period ;the differences in the two indicators mentioned above among the 3 groups were compared with the control group; the positive rates for WBC and CRP were calculated and compared in accordance with its corresponding positive diagnostic criteria! the value of CRP in the diagnosis of pneumonia in children was comprehensively evaluated. RESULTS The study showed that WBC of the bacterial pneumonia group, viral pneumonia group, children with Mycoplasma pneumonia group, and the control group, as receiving the treatment, were (11.7 ± 4. 1),<9. 5±3. 0),(9. 0±3.1), and (7. 0±2. 2) X109/L; CRP were respectively (52. 3±24. 0) ,(9. 05 ±3. 3), (34. 7 ± 13.4), and (3. 9 ± 2. 1) mg/L ;both CRP and WMC of bacterial pneumonia group, viral pneumonia, Mycoplasma pneumonia group were significantly higher than those of the control group) the CRP and WBC of bacterial pneumonia group were significantly higher than those of viral pneumonia group and Mycoplasma pneumonia group, the difference was statistically significant (P<0. 05). the positive rates of WBC of the children in bacterial pneumonia group, viral pneumonia group and Mycoplasma pneumonia group were 81. 3%, 77. 5% , and 66. 5%, respectively, the positive rates of CRP were 100.0%, 74. 5%, and 73. i

  13. Evaluation of the persistence and gene expression of an anti-Chlamydophila psittaci DNA vaccine in turkey muscle

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    Vanrompay Daisy

    2006-06-01

    Full Text Available Abstract Background DNA vaccination has been shown to elicit specific cellular and humoral immune responses to many different agents in a broad variety of species. However, looking at a commercial use, the duration of the immune response against the vaccine is critical. Therefore the persistence of the DNA vaccine, as well as its expression, should be investigated. We conducted these investigations on a DNA vaccine against Chlamydophila psittaci, a Gram-negative intracellular bacterium which causes respiratory disease in turkeys and humans. Previous studies showed that the DNA vaccine confers partial protection against C. psittaci infection in turkeys. Turkeys were injected intramuscularly with the DNA vaccine : a eukaryotic expression vector (pcDNA1::MOMP expressing the major outer membrane protein (MOMP of an avian C. psittaci serovar D strain. Over a period of 11 weeks, cellular uptake of the DNA vaccine was examined by PCR, transcription of the insert by reverse transcript-PCR (RT-PCR and mRNA translation by immunofluorescence staining of muscle biopsies. Results The results indicate that the DNA vaccine persists in turkey muscle for at least 10 weeks. Moreover, during this period of time MOMP was continuously expressed, as evidenced by the immunofluorescence staining and RT-PCR. Conclusion Since C. psittaci infections occur at the age of 3 to 6 and 8 to 12 weeks, a vaccine persistence of 10 weeks seems adequate. Therefore, further research should concentrate on improving the elicited immune response, more specifically the cell-mediated immune response, rather than prolonging the lifespan of the plasmid.

  14. Use of a nested PCR-enzyme immunoassay with an internal control to detect Chlamydophila psittaci in turkeys

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    Goddeeris Bruno M

    2005-09-01

    Full Text Available Abstract Background Laboratory diagnosis of Chlamydophila psittaci, an important turkey respiratory pathogen, is difficult. To facilitate the diagnosis, a nested PCR-enzyme immunoassay (PCR-EIA was developed to detect the Cp. psittaci outer membrane protein A (ompA gene in pharyngeal swabs. Methods The fluorescein-biotin labelled PCR products were immobilized on streptavidin-coated microtiter plates and detected with anti-fluorescein peroxidase conjugate and a colorimetric substrate. An internal inhibition control was included to rule out the presence of inhibitors of DNA amplification. The diagnostic value of the ompA nested PCR-EIA in comparison to cell culture and a 16S-rRNA based nested PCR was assessed in pharyngeal turkey swabs from 10 different farms experiencing respiratory disease. Results The sensitivity of the nested PCR-EIA was established at 0.1 infection forming units (IFU. Specificity was 100%. The ompA nested PCR-EIA was more sensitive than the 16S-rRNA based nested PCR and isolation, revealing 105 out of 200 (52.5% positives against 13 and 74 for the latter two tests, respectively. Twenty-nine (23.8% out of 122 ompA PCR-EIA negatives showed the presence of inhibitors of DNA amplification, although 27 of them became positive after diluting (1/10 the specimens in PCR buffer or after phenol-chloroform extraction and subsequent ethanol precipitation. Conclusion The present study stresses the need for an internal control to confirm PCR true-negatives and demonstrates the high prevalence of chlamydiosis in Belgian turkeys and its potential zoonotic risk. The ompA nested PCR-EIA described here is a rapid, highly sensitive and specific diagnostic assay and will help to facilitate the diagnosis of Cp. psittaci infections in both poultry and man.

  15. GroEL1, a heat shock protein 60 of Chlamydia pneumoniae, impairs neovascularization by decreasing endothelial progenitor cell function.

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    Yi-Wen Lin

    Full Text Available The number and function of endothelial progenitor cells (EPCs are sensitive to hyperglycemia, hypertension, and smoking in humans, which are also associated with the development of atherosclerosis. GroEL1 from Chlamydia pneumoniae has been found in atherosclerotic lesions and is related to atherosclerotic pathogenesis. However, the actual effects of GroEL1 on EPC function are unclear. In this study, we investigate the EPC function in GroEL1-administered hind limb-ischemic C57BL/B6 and C57BL/10ScNJ (a toll-like receptor 4 (TLR4 mutation mice and human EPCs. In mice, laser Doppler imaging, flow cytometry, and immunohistochemistry were used to evaluate the degree of neo-vasculogenesis, circulating level of EPCs, and expression of CD34, vWF, and endothelial nitric oxide synthase (eNOS in vessels. Blood flow in the ischemic limb was significantly impaired in C57BL/B6 but not C57BL/10ScNJ mice treated with GroEL1. Circulating EPCs were also decreased after GroEL1 administration in C57BL/B6 mice. Additionally, GroEL1 inhibited the expression of CD34 and eNOS in C57BL/B6 ischemic muscle. In vitro, GroEL1 impaired the capacity of differentiation, mobilization, tube formation, and migration of EPCs. GroEL1 increased senescence, which was mediated by caspases, p38 MAPK, and ERK1/2 signaling in EPCs. Furthermore, GroEL1 decreased integrin and E-selectin expression and induced inflammatory responses in EPCs. In conclusion, these findings suggest that TLR4 and impaired NO-related mechanisms could contribute to the reduced number and functional activity of EPCs in the presence of GroEL1 from C. pneumoniae.

  16. Prevalence and risk factors associated with Chlamydophila abortus infection in dairy herds in Jordan.

    Science.gov (United States)

    Talafha, Abdelsalam Q; Ababneh, Mohammed M; Ababneh, Mustafa M; Al-Majali, Ahmad M

    2012-12-01

    A cross-sectional study was carried out to determine seroprevalence and to identify risk factors associated with Chlamydophila abortus infection in 62 nonvaccinated dairy herds (671 cows) in Jordan between January and June 2007. Information regarding herd management was recorded through a personal interview with farmers. Antibodies against C. abortus were detected using an ELISA test kit. Chi-square analysis and multivariable logistic regression model were used to identify risk factors associated with C. abortus seropositivity. The true prevalence of antibodies against C. abortus in individual cows and cattle herds were 19.9 % and 66.3 %, respectively. Univariable Chi-square analysis revealed three variables with P ≤ 0.25 that were further offered to multivariable logistic regression analysis. Small-sized herds were identified as a risk factor for seropositivity to C. abortus, while sweeping followed by water hosing and using disinfectants were identified as protective factors. Cows in the age groups of >8 and ≤ 10 years old and >2 and ≤ 6 years old had the highest and lowest significant seroprevalence to C. abortus, respectively. Results of this study indicated that C. abortus is highly prevalent in Jordan's dairy herds and Chlamydophila infection could be controlled by applying strict biosecurity measures in the dairy farms.

  17. Investigation of Chlamydiaceae in semen and cauda epididymidis and seroprevalence of Chlamydophila abortus in breeding bulls

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    Persson Ylva

    2010-01-01

    Full Text Available Abstract Background Reproductive disorders associated with chlamydial infection have been reported worldwide in cattle and there are indications of potential venereal transmission. Methods Semen samples from 21 dairy bulls and cauda epididymidis tissue samples from 43 beef bulls were analysed for chlamydial agent by real-time polymerase chain reaction (PCR including an internal amplification control (mimic. Additionally, presence of antibodies against Chlamydophila (Cp. abortus among the bulls was investigated with the commercial Pourquier® ELISA Cp. abortus serum verification kit. Results No chlamydial agent was detected by PCR in either the semen samples or in the tissue samples. Additionally, no antibodies against Cp. abortus were detected. Conclusions The results suggest that Cp. abortus is very rare, or absent in Swedish bulls and thus the risk for venereal transmission of chlamydial infection through their semen is low. However, because Chlamydophila spp. infection rates seem to differ throughout the world, it is essential to clarify the relative importance of transmission of the infection through semen on cattle fertility.

  18. Mixed infection with reovirus and Chlamydophila in a flock of budgerigars (Melopsittacus undulatus).

    Science.gov (United States)

    Perpiñán, David; Garner, Michael M; Wellehan, James F X; Armstrong, Douglas L

    2010-12-01

    Eleven budgerigars (Melopsittacus undulatus) from a zoological collection presented at necropsy with emaciation and splenomegaly or hepatomegaly or both. Polymerase chain reaction assays performed on liver and spleen samples were positive for Chlamydophila psittaci in 2 of 3 birds tested, and histologic findings in 2 additional birds were compatible with chlamydiosis. The aviary was subsequently closed to the public, and a 45-day treatment regimen with doxycycline in the seeds was initiated. No further deaths of birds with hepatomegaly or splenomegaly were observed after the first day of treatment. Further investigation of birds that died during the outbreak with emaciation and with hepatic and splenic enlargement revealed severe necrosis of the spleen and liver suggestive of reovirus infection, which was supported with polymerase chain reaction analysis from paraffin-embedded tissue. This reovirus was sequenced and had 100% homology with a reovirus previously identified in an African grey parrot (Psittacus erithacus). The outbreak did not affect cockatiels (Nymphicus hollandicus) and blue quail (Coturnix chinensis) kept in the same aviary. A group of budgerigars added to the collection soon before the opening of the aviary may have introduced reovirus and Chlamydophila into the collection.

  19. 血清C反应蛋白检测在小儿支原体肺炎感染中的临床价值%Value of C-reactive Protein Detection in Children with Mycoplasma Pneumonia Infection

    Institute of Scientific and Technical Information of China (English)

    左克

    2012-01-01

    Objective: To explore the diagnostic value of C-reactive protein ( CRP ) in infant with my-coplasma pneumonia. Method: The serum CRP levels were measured by nephelometric immunoassay and ELISA in 98 infants with mycoplasma pneumonia ( mycoplasma pneumonia group ) and 67 infants with bacterial pneumonia ( bacterial pneumonia group ) 72 healthy children ( control group ). Result: Mean CRP levels of the pneumonia group and bacterial pneumonia group on admission were signifcantly increased ( P mycoplasma pneumonia group ( P0. 05 ), however, the bacterial pneumonia group's CRP level was still higher than mycoplasma pneumonia group' s and the control group' s ( P< 0. 05 ); Among 115 childern whose CRP level were increased in admission, 68 children ( 59. 13% ) were younger than 4 years old, 47 children ( 40. 87% ) were 4 years old or older than 4, and the youngest of them was only 2 months. Conclusion: The CRP level for children with mycoplasma pneumonia is slightly higher than normal value, but lower than that of bacterial pneumonia. CRP can be used as an important auxiliary index of identification of mycoplasma pneumonia or bacterial pneumonia.%目的 探讨血清C反应蛋白检测在小儿支原体肺炎感染中的临床价值.方法 采用免疫比浊法测定98例支原体肺炎(支原体肺炎组)和67例细菌性肺炎患儿(细菌性肺炎组)血清 CRP,与 72例健康儿童(健康对照组)作对比.结果 入院时支原体肺炎组和细菌性肺炎组CRP均值和阳性率均显著高于对照组(P<0.05),且细菌性肺炎组显著高于支原体肺炎组(P<0.05);治疗1周后,两组CRP和阳性率指标较入院时均明显下降,支原体肺炎组与健康对照组比较无差别(P>0.05),但细菌性肺炎组仍高于支原体肺炎组和健康对照组(P<0.05);所有病例在入院时CRP和MP-IgM检测为阳性者,4岁以下儿童为59.13%(68/115),4岁以上为40.87%(47/115),其中年龄最小的仅生后2个月.结论 对于支原体肺炎患

  20. Comparison of polymerase chain reaction tests for diagnosis of feline herpesvirus, Chlamydophila felis, and Mycoplasma spp. infection in cats with ocular disease in Canada.

    Science.gov (United States)

    Sandmeyer, Lynne S; Waldner, Cheryl L; Bauer, Bianca S; Wen, Xin; Bienzle, Dorothee

    2010-06-01

    This study assessed the value of polymerase chain reaction (PCR) for making a diagnosis of feline herpesvirus (FHV-1) infection, and for differentiating this from Chlamydophila felis and Mycoplasma spp. infection in a clinical setting in Canada. We compared the frequency of positive FHV-1 PCR test results from 48 clinical cases of ocular disease in cats suspected to be due to FHV-1 between 1 research and 2 commercial laboratories in Canada. We also compared PCR results for Chlamydophila felis and Mycoplasma spp. between the 2 commercial laboratories. The prevalence of FHV-1 infection in the cats ranged from 4% to 21%. The prevalence of Chlamydophila felis was 2% and 17% and the prevalence of Mycoplasma spp. was 11% and 27%. Agreement between FHV-1 culture and PCR results at the research laboratory was substantial (kappa = 0.76). There was slight agreement (kappa Chlamydophila felis (kappa = 0.2) and Mycoplasma spp. (kappa = 0.07) PCR.

  1. Intranasal Immunization with the Cholera Toxin B Subunit-Pneumococcal Surface Antigen A Fusion Protein Induces Protection against Colonization with Streptococcus pneumoniae and Has Negligible Impact on the Nasopharyngeal and Oral Microbiota of Mice

    OpenAIRE

    F.C. Pimenta; Miyaji, E. N.; Arêas, A. P. M.; Oliveira, M. L. S.; de Andrade, A. L. S. S.; Ho, P.L.; Hollingshead, S. K.; Leite, L. C. C.

    2006-01-01

    One of the candidate proteins for a mucosal vaccine antigen against Streptococcus pneumoniae is PsaA (pneumococcal surface antigen A). Vaccines targeting mucosal immunity may raise concerns as to possible alterations in the normal microbiota, especially in the case of PsaA, which was shown to have homologs with elevated sequence identity in other viridans group streptococci. In this work, we demonstrate that intranasal immunization with a cholera toxin B subunit-PsaA fusion protein is able to...

  2. Chlamydophila species in dairy farms: polymerase chain reaction prevalence, disease association, and risk factors identified in a cross-sectional study in western Germany.

    Science.gov (United States)

    Kemmerling, K; Müller, U; Mielenz, M; Sauerwein, H

    2009-09-01

    The prevalence of Chlamydophila spp. was determined in a cross-sectional study carried out in 2007 using 100 randomly selected dairy herds in the western part of Germany. Ten dairy cows per herd were sampled in herds with fewer than 100 cows; in bigger herds, 10% of the cows were sampled. For the detection of Chlamydophila spp., vaginal swabs from early lactating dairy cows were analyzed using an established highly sensitive genus-specific real-time PCR. In consideration of the discontinuous shedding of the pathogen, a herd was classified as positive if at least 1 animal per herd tested positive for Chlamydophila spp. By use of these methods and definitions, 61% of the dairy herds and 13.5% of the cows were detected as PCR-positive for Chlamydophila spp., which is indicative for ongoing infections. To compare herd health and herd performance between herds testing positive or negative and to identify risk factors for the presence of Chlamydophila spp., a questionnaire was designed to evaluate farm characteristics and management practices. In addition, the performance recordings of the state dairy recording organization were used for these purposes. Milk yield, number of lactations, and calving to first-service interval were lower in herds testing positive for Chlamydophila spp. compared with negative herds. For all these variables, there was no interaction between Chlamydophila status and lactation number. Replacement of animals from outside sources, use of breeding bulls, lack of separate calving pens, and low scores for cleanliness of beddings, walkways, and cows were identified as the main risk factors for Chlamydophila spp.

  3. Sprouty-Related Ena/Vasodilator-Stimulated Phosphoprotein Homology 1-Domain-Containing Protein-2 Critically Regulates Influenza A Virus-Induced Pneumonia.

    Science.gov (United States)

    Ito, Toshihiro; Itakura, Junya; Takahashi, Sakuma; Sato, Miwa; Mino, Megumi; Fushimi, Soichiro; Yamada, Masao; Morishima, Tuneo; Kunkel, Steven L; Matsukawa, Akihiro

    2016-07-01

    Influenza A virus causes acute respiratory infections that induce annual epidemics and occasional pandemics. Although a number of studies indicated that the virus-induced intracellular signaling events are important in combating influenza virus infection, the mechanism how specific molecule plays a critical role among various intracellular signaling events remains unknown. Raf/MEK/extracellular signal-regulated kinase cascade is one of the key signaling pathways during influenza virus infection, and the Sprouty-related Ena/vasodilator-stimulated phosphoprotein homology 1-domain-containing protein has recently been identified as a negative regulator of Raf-dependent extracellular signal-regulated kinase activation. Here, we examined the role of Raf/MEK/extracellular signal-regulated kinase cascade through sprouty-related Ena/vasodilator-stimulated phosphoprotein homology 1-domain-containing protein in influenza A viral infection because the expression of sprouty-related Ena/vasodilator-stimulated phosphoprotein homology 1-domain-containing protein was significantly enhanced in human influenza viral-induced pneumonia autopsy samples. Prospective animal trial. Research laboratory. Wild-type and sprouty-related Ena/vasodilator-stimulated phosphoprotein homology 1-domain-containing protein-2 knockout mice inoculated with influenza A. Wild-type or sprouty-related Ena/vasodilator-stimulated phosphoprotein homology 1-domain-containing protein-2 knockout mice were infected by intranasal inoculation of influenza A (A/PR/8). An equal volume of phosphate-buffered saline was inoculated intranasally into mock-infected mice. Influenza A infection of sprouty-related Ena/vasodilator-stimulated phosphoprotein homology 1-domain-containing protein-2 knockout mice led to higher mortality with greater viral load, excessive inflammation, and enhanced cytokine production than wild-type mice. Administration of MEK inhibitor, U0126, improved mortality and reduced both viral load and

  4. Difference of clinical features in childhood Mycoplasma pneumoniae pneumonia

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    Kang Jin-Han

    2010-07-01

    Full Text Available Abstract Background M. pneumoniae pneumonia (MP has been reported in 10-40% of community-acquired pneumonia cases. We aimed to evaluate the difference of clinical features in children with MP, according to their age and chest radiographic patterns. Methods The diagnosis of MP was made by examinations at both admission and discharge and by two serologic tests: the indirect microparticle agglutinin assay (≥1:40 and the cold agglutinins titer (≥1:32. A total of 191 children with MP were grouped by age: ≤2 years of age (29 patients, 3-5 years of age (81 patients, and ≥6 years of age (81 patients. They were also grouped by pneumonia pattern: bronchopneumonia group (96 patients and segmental/lobar pneumonia group (95 patients. Results Eighty-six patients (45% were seroconverters, and the others showed increased antibody titers during hospitalization. Among the three age groups, the oldest children showed the longest duration of fever, highest C-reactive protein (CRP values, and the most severe pneumonia pattern. The patients with segmental/lobar pneumonia were older and had longer fever duration and lower white blood cell (WBC and lymphocyte counts, compared with those with bronchopneumonia. The patient group with the most severe pulmonary lesions had the most prolonged fever, highest CRP, highest rate of seroconverters, and lowest lymphocyte counts. Thrombocytosis was observed in 8% of patients at admission, but in 33% of patients at discharge. Conclusions In MP, older children had more prolonged fever and more severe pulmonary lesions. The severity of pulmonary lesions was associated with the absence of diagnostic IgM antibodies at presentation and lymphocyte count. Short-term paired IgM serologic test may be mandatory for early and definitive diagnosis of MP.

  5. Clinical characteristics of pulmonary embolism with concomitant pneumonia.

    Science.gov (United States)

    Cha, Seung-Ick; Choi, Keum-Ju; Shin, Kyung-Min; Lim, Jae-Kwang; Yoo, Seung-Soo; Lee, Jaehee; Lee, Shin-Yup; Kim, Chang-Ho; Park, Jae-Yong

    2016-04-01

    Although pneumonia is associated with an increased risk of venous thromboembolism, patients with pulmonary embolism and concomitant pneumonia are uncommon. The aim of the present study was to investigate the clinical features of pulmonary embolism with coexisting pneumonia. We retrospectively compared clinical, radiologic and laboratory parameters between patients with pulmonary embolism and concomitant pneumonia (pneumonia group) and those with unprovoked pulmonary embolism (unprovoked group), and then between the pneumonia group and those with pulmonary infarction (infarction group). Of 794 patients with pulmonary embolism, 36 (5%) had coexisting pneumonia and six (1%) had no provoking factor other than pneumonia. Stroke was significantly more common in the pneumonia group, than either the unprovoked group or the infarction group. In the pneumonia group, fever was significantly more common and serum C-reactive protein levels were significantly higher. By contrast, central pulmonary embolism and right ventricular dilation on computed tomography were significantly less frequent in the pneumonia group. In addition, an adverse outcome due to pulmonary embolism was less common in the pneumonia group than in either of the other two groups. The coexistence of pulmonary embolism and pneumonia is rarely encountered in clinical practice, especially without the presence of other factors that could provoke venous thromboembolism and is commonly associated with stroke. It is characterized by lower incidences of central pulmonary embolism and right ventricular dilation and by a lower rate of adverse outcomes due to pulmonary embolism itself.

  6. Childhood pneumonia and vitamin A

    Directory of Open Access Journals (Sweden)

    Farhad Heidarian

    2014-04-01

    Full Text Available One of the major causes of mortality in children younger than 5 years old is acute lower respiratory tract infections (ALRI. ALRI clinical features are cough, tachypnea, fever, coryza, chest retraction, crackles and wheeze. Increased white blood cell count with left shift might happen in pneumonia. C-reactive protein (CRP and erythrocyte sedimentation rate (ESR might rise in children with respiratory tract infections. Vitamin A deficiency is associated with severe childhood infections. The effect of vitamin A supplementation in childhood pneumonia depends on the prevalence and the level of vitamin A deficiency in the population. Some studies confirmed that retinol levels were significantly higher after recovery from acute pneumonia compared to acute phase. But there were no significant association between serum retinol level and the clinical manifestation.

  7. Experimentally produced calf pneumonia.

    Science.gov (United States)

    Gourlay, R N; Howard, C J; Thomas, L H; Stott, E J

    1976-03-01

    Experimental pneumonia was produced in calves by the endobronchial inoculation of pneumonic lung homogenates. Irradiated homogenates produced minimal pneumonia. Ampicillin treatment of the homogenates and the experimental calves reduced the extent of pneumonia. Treatment with tylosin tartrate prevented experimental pneumonia. These results suggest that the total pneumonia was due to organisms susceptible to tylosin tartrate and that the residual pneumonia remaining after ampicillin treatment was due to organisms susceptible to tylosin tartrate but not to ampicillin. Of the organisms isolated from the lungs, the ones in this latter category most likely to be responsible are Mycoplasma dispar and ureaplasmas (T-mycoplasmas).

  8. [Community acquired pneumonia in children: an update for outpatients management].

    Science.gov (United States)

    Wagner, Noémie; Gehri, Mario; Gervaix, Alain; Guinan, Stéphane; Barazzone-Argiroffo, Constance

    2016-02-17

    Pneumonia should be considered in febrile children with tachypnea and/or chest recession. Virus are the most common cause of pneumonia in children under 5 years old. Streptococcus pneumonia can be found at any age. Mycoplasma pneumonia is more frequent in older children. Systematic chest radiograph is not necessary but must be obtained in patients with hypoventilation and in those with failed initial antibiotic therapy. Mycoplasma pneumonia should be tested according to patient age and response to initial antibiotic. First line antibiotherapy is amoxicilline. Antibiotic treatment is frequently not necessary in children under 5 but should be considered depending on clinical presentation and C reactive protein value.

  9. Confirmation of Chlamydophila abortus in infected cell culture using Indirect Immunofluorescence technique

    Directory of Open Access Journals (Sweden)

    Krishnan Nair G

    Full Text Available Chlamydophila abortus (C. abortus is an important abortifacient agent in bovines and ovines. Clinical diagnosis of the disease is often difficult. An early diagnosis can be achieved based on direct demonstration of the organism in clinical material and through the cultural recovery of the organism in embryonated chicken egg. For confirmatory diagnosis antigen detection methods or serological techniques can be adopted. The present study is aimed at the confirmatory diagnosis of C. abortus infection by indirect immunofluorescence technique following the isolation of the organism in cell culture. Specific apple green fluorescing inclusions of C. abortus in McCoy cell lines was detected from 72 h to 96 h post infection employing anti-chlamydial group specific monoclonal antibodies. Thus, a confirmatory diagnosis of the infection was possible with this study. [Vet. World 2011; 4(10.000: 473-474

  10. Mixed infection by fowlpox virus and Chlamydophila psittaci in a commercial laying hen flock.

    Science.gov (United States)

    Karpińska, Teresa Agnieszka; Kozaczyński, Wojciech; Niemczuk, Krzysztof; Jasik, Agnieszka; Kycko, Anna; Reichert, Michał

    2014-03-01

    An outbreak of fowlpox occurred in a commercial laying hen flock in one of the western provinces of Poland. Clinical signs suggested fowlpox and the diagnosis was confirmed by histopathological detection of Bollinger bodies within the epithelial cells. Detailed ultrastructural examination revealed an additional concurrent infection with chlamydia-like particles. The particles were identified by PCR as fowlpox virus and Chlamydophila psittaci. It is worth noting that both pathogens can generate morphologic forms capable of prolonged survival and inducing latent and persistent infection. We suggest a possible interaction between the two pathogens on ultrastructural level and assess the clinical consequences of the mixed infection. This study also demonstrates a potential of the transmission electron microscope (TEM) for identifying a superinfection with another pathogen (in this case C. psittaci), which may remain undetected by routine techniques.

  11. Chlamydophila abortus infection in the mouse: a useful model of the ovine disease.

    Science.gov (United States)

    Caro, M R; Buendía, A J; Del Rio, L; Ortega, N; Gallego, M C; Cuello, F; Navarro, J A; Sanchez, J; Salinas, J

    2009-03-16

    Chlamydophila (C.) abortus is an obligate intracellular bacterium able to colonize the placenta of several species of mammals, which may induce abortion in the last third of pregnancy. The infection affects mainly small ruminants resulting in major economic losses in farming industries worldwide. Furthermore, its zoonotic risk has been reported in pregnant farmers or abattoir workers. Mouse models have been widely used to study both the pathology of the disease and the role of immune cells in controlling infection. Moreover, this animal experimental model has been considered a useful tool to evaluate new vaccine candidates and adjuvants that could prevent abortion and reduce fetal death. Future studies using these models will provide and reveal information about the precise mechanisms in the immune response against C. abortus and will increase the knowledge about poorly understood issues such as chlamydial persistence.

  12. Efficacy of a new inactivated Chlamydophila felis vaccine in experimentally-infected cats.

    Science.gov (United States)

    Masubuchi, Katsuo; Wakatsuki, Akira; Iwamoto, Kayo; Takahashi, Takuo; Kokubu, Teruaki; Shimizu, Mitsugu

    2010-08-01

    A new inactivated and adjuvanted Chlamydophila felis vaccine was developed and its efficacy in cats was compared with that of commercially available inactivated and live vaccines. Two commercial vaccines conferred insufficient immunity on inoculated cats, as evaluated by antibody production and a challenge experiment, whereas cats administered the newly generated vaccine produced high-titre antibodies and acquired sufficient immunity. The cats immunised with the new vaccine revealed no or only mild clinical signs, and no chlamydiae were recovered from their tissue samples after exposure to a virulent C felis. However, they shed chlamydiae in their nasal and conjunctival secretions after challenge, as did those immunised with the commercial vaccines and the non-vaccinated controls. The newly developed vaccine caused no adverse reaction in the inoculated cats. These findings suggest that the new vaccine prepared here may be promising for practical use in controlling C felis infection in cats.

  13. Salmonella, Campylobacter, and Chlamydophila in bald ibis (Geronticus eremita) feces in Turkey.

    Science.gov (United States)

    Tel, Osman Yaşar; Bozkaya, Faruk; Keskin, Oktay

    2013-03-01

    The aim of this study was to investigate the presence of Campylobacter spp., Salmonella spp., and Chlamydophila psittaci in fecal samples of bald ibises (Geronticus eremita) housed in a conservation facility in Turkey. A total of 82 fecal samples were collected from cages and evaluated by bacteriologic methods and a polymerase chain reaction (PCR) technique for Campylobacter spp. and Salmonella spp. and by PCR for C. psittaci. Campylobacter spp. were isolated from 24 of 82 fecal samples (29.2%). Of these 18 (75%), 4 (16.7%) and 2 (8.3%) were Campylobacter jejuni, Campylobacter coli, and other Campylobacter spp., respectively. Salmonella spp. were detected in 8 fecal specimens.(9.7%) by PCR. The presence of C. psittaci was not detected in the bald ibises studied. The results suggested that the bald ibises in this present study might be at a higher risk of infection with Salmonella spp. and Campylobacter spp.

  14. Increasing effect of a high dose of PG-1 peptide on the infectivity of Chlamydophila abortus.

    Science.gov (United States)

    Donati, Manuela; Di Francesco, Antonietta; Gennaro, Renato; Benincasa, Monica; Di Paolo, Maria; Shurdhi, Alisa; Ostanello, Fabio; Baldelli, Raffaella; Cevenini, Roberto

    2010-07-01

    Cathelicidins are antimicrobial peptides, stored by mammalian leukocytes, showing an antimicrobial activity against bacteria, fungi, protozoa and enveloped viruses. In accordance with other authors, we reported in a previous study that the protegrin-1 (PG-1), at 80 microg mL(-1), inhibited the in vitro growth of Chlamydia trachomatis serovars D, H and L2; however, we observed an increased infectivity of some animal chlamydial species after their treatment with the same PG-1 concentration. In this study, the treatment of LLC-MK2 cells with PG-1 before chlamydial infection resulted in an increased infectivity of Chlamydophila abortus probably due to their easier entry into the host cells, whereas no increase in S26/3 infectivity was detected in LLC-MK2 cells treated with PG-1 postchlamydial infection.

  15. Seroprevalence of antibodies to Chlamydophila abortus in ovine in the State of Alagoas, Brazil

    Directory of Open Access Journals (Sweden)

    José Wilton Pinheiro Junior

    2010-06-01

    Full Text Available The goal of this study was to perform a seroepidemiological investigation and to identify risk factors associated with infection of Chlamydophila abortus of sheep herds in the Brazilian state of Alagoas. The study was conducted with samples of 274 ewes with ages equal to or higher than 24 months in 25 herds and in 23 towns located in three regions of the state (Sertão, Agreste and Eastern Alagoas. Anti-C. abortus antibodies were detected using the microcomplement fixation test. The risk factors, were determined based on questionnaires consisting of objective questions, about the farmer and general characteristics of the herd like size, sanitary situation and reproductive management. Among 274 sera samples analyzed for C. abortus, 59 (21.5% were positive with titers >32, 187 (68.3% negative and 28 (10.2% suspect with titers >16. In the 23 towns studied, 20 had positive animals. Among herds 21 (77.7% of had positive animals. The only variable which appeared to be significant in the multivariate analysis was the region, and Sertão was the most significant (p<0.001; OR=3.48; T.I. 1.79 - 6.76. Results indicate that infection by Chlamydophila abortus is widespread on sheep farms in the State of Alagoas. Others studies, however, have to be conducted to isolate the agent in order to confirm the role of the bacteria is reproductive disturbances in sheeps. In addition to that, control and prophylactic measures along with health promoting programs have to be encouraged on the studied farms so that infection reates are reduced.

  16. Serum Antibody Response to Five Streptococcus pneumoniae Proteins during Acute Otitis Media in Otitis Prone and Non-Otitis Prone Children

    Science.gov (United States)

    Kaur, Ravinder; Casey, Janet R.; Pichichero, Michael E.

    2011-01-01

    Background Streptococcus pneumoniae (Spn) is one of the common bacteria responsible for episodic acute otitis media (AOM; non-otitis prone), recurrent AOM (otitis-prone) and AOM treatment failure (AOMTF) in children. Objective From a population of 268 children we sought to compare the serum IgG antibody titers to five different Spn proteins (PhtD, LytB, PcpA, PhtE and Ply) that are vaccine candidates in children with episodic AOM (n=34), who were otitis prone (n=35), and who had AOMTF (n=25) caused by Spn. Methods Antibody was quantitated by ELISA. Results At their acute AOM visit, anti-PhtD, -LytB, -PhtE and −Ply IgG antibody titers in otitis-prone children were significantly lower compared to non-otitis prone children (p otitis-prone, AOMTF and non-otitis prone children had no significant change in geometric mean IgG antibody titers against the five proteins (except for PhtE in children with AOMTF), but detailed analysis showed that about one-third of the children in each cohort had a 2-fold rise in antibody to the studied antigens. While non-otitis prone children had significant increases (p otitis-prone children either failed to show rises or the rises were significantly less than the non-otitis prone children. Conclusion Otitis-prone and AOMTF children mount less of an IgG serum antibody response than non-otitis prone children to Spn proteins following AOM and nasopharyngeal colonization. PMID:21487325

  17. YjcC, a c-di-GMP phosphodiesterase protein, regulates the oxidative stress response and virulence of Klebsiella pneumoniae CG43.

    Directory of Open Access Journals (Sweden)

    Ching-Jou Huang

    Full Text Available This study shows that the expression of yjcC, an in vivo expression (IVE gene, and the stress response regulatory genes soxR, soxS, and rpoS are paraquat inducible in Klebsiella pneumoniae CG43. The deletion of rpoS or soxRS decreased yjcC expression, implying an RpoS- or SoxRS-dependent control. After paraquat or H2O2 treatment, the deletion of yjcC reduced bacterial survival. These effects could be complemented by introducing the ΔyjcC mutant with the YjcC-expression plasmid pJR1. The recombinant protein containing only the YjcC-EAL domain exhibited phosphodiesterase (PDE activity; overexpression of yjcC has lower levels of cyclic di-GMP. The yjcC deletion mutant also exhibited increased reactive oxygen species (ROS formation, oxidation damage, and oxidative stress scavenging activity. In addition, the yjcC deletion reduced capsular polysaccharide production in the bacteria, but increased the LD50 in mice, biofilm formation, and type 3 fimbriae major pilin MrkA production. Finally, a comparative transcriptome analysis showed 34 upregulated and 29 downregulated genes with the increased production of YjcC. The activated gene products include glutaredoxin I, thioredoxin, heat shock proteins, chaperone, and MrkHI, and proteins for energy metabolism (transporters, cell surface structure, and transcriptional regulation. In conclusion, the results of this study suggest that YjcC positively regulates the oxidative stress response and mouse virulence but negatively affects the biofilm formation and type 3 fimbriae expression by altering the c-di-GMP levels after receiving oxidative stress signaling inputs.

  18. Evaluation of Chlamydophila abortus DNA extraction protocols for polymerase chain reaction diagnosis in paraffin-embedded tissues.

    Science.gov (United States)

    Ortega, Nieves; Navarro, José A; Nicolás, Laura; Buendía, Antonio J; Caro, María R; Del Río, Laura; Martínez, Carlos M; Cuello, Francisco; Salinas, Jesús; Gallego, María C

    2007-07-01

    Polymerase chain reaction (PCR) has gained increasing importance as a tool for directly demonstrating the presence of Chlamydophila in the placentas of aborted sheep and goats. However, because of the zoonotic potential of the disease, it is advisable to use fixed materials. To evaluate 4 different DNA extraction protocols in paraffin-embedded sections for PCR, previously immunohistochemically diagnosed placental samples from outbreaks of abortions in goats and sheep were used. The samples were also used to evaluate the effect of the duration of fixation in formalin on PCR. A protocol that uses Tris-HCl pH 8.5 with EDTA and subsequent digestion with proteinase K was found to be an easy protocol for obtaining excellent PCR products for Chlamydophila abortus diagnosis from formalin-fixed and paraffin-embedded specimens. It was also found that if samples are fixed in formalin for more than 2 weeks, the PCR technique is affected more adversely than immunohistochemical methods.

  19. Prevalence of antibodies to Chlamydophila abortus in ovines in the Londrina area of Paraná state, Brazil

    Directory of Open Access Journals (Sweden)

    Bruna Azevedo de Carvalho Lima

    2014-10-01

    Full Text Available The aim of this paper was to estimate the prevalence of antibodies to Chlamydophila abortus in ewes from the Londrina region of Paraná state. Blood samples from 267 adult ewes from eight herds were submitted to complement fixation testing. Tests were considered positive when the titers ? 32. Nineteen animals (7.11% in 7 (87.50% herds tested positive for antibodies against C. abortus. The titers were 32 in 17 animals and were 64 and 128 in the other two animals. The clinical signs recorded were abortion, stillbirth, weak newborns and heat repetition. The high frequency of herds with at least one positive animal and a seroprevalence of 7.11% suggest the presence of Chlamydophila abortus in the animals of studied herds.

  20. Investigation of the possible role of Chlamydophila abortus in reproductive failures in nrazilian herds of domestic ruminants

    OpenAIRE

    Francielle Gibson da Silva-Zacarias; Amauri Alcindo Alfieri; Kledir Anderson Hofstaetter Spohr; Bruna Azevedo de Carvalho Lima; Rosângela Claret de Oliveira; Carlo Turilli; Michele Lunardi; Rodrigo Alejandro Arellano Otonel; Julio Cesar de Freitas

    2009-01-01

    Chlamydophila abortus (C. abortus) infection is related to reproductive failure in domestic ruminants. Although it has not been well characterized worldwide, this pathogen has already been identified in some European countries and in the USA. In Brazil, preliminary studies have shown serological evidence of C. abortus infection in herds with low antibody prevalence. Until now, the identification of C. abortus in biological samples from females presenting reproductive failures has not been des...

  1. Pneumonia - children - discharge

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    ... this page: //medlineplus.gov/ency/patientinstructions/000011.htm Pneumonia in children - discharge To use the sharing features ... this page, please enable JavaScript. Your child has pneumonia, which is an infection in the lungs. In ...

  2. Pneumonia - adults - discharge

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    ... this page: //medlineplus.gov/ency/patientinstructions/000017.htm Pneumonia in adults - discharge To use the sharing features on this page, please enable JavaScript. You have pneumonia, which is an infection in your lungs. In ...

  3. Pneumocystis jiroveci pneumonia

    Science.gov (United States)

    ... medlineplus.gov/ency/article/000671.htm Pneumocystis jiroveci pneumonia To use the sharing features on this page, please enable JavaScript. Pneumocystis jiroveci pneumonia is a fungal infection of the lungs. The ...

  4. NEW VIRULENCE FACTORS OF STREPTOCOCCUS PNEUMONIAE

    NARCIS (Netherlands)

    Hermans, Peter Wilhelmus Maria; Bootsma, Jeanette Hester; Burghout, Pieter Jan; Kuipers, Oscar; Bijlsma, Johanna Jacoba Elisabeth; Kloosterman, Tomas Gerrit; Andersen, Christian O.

    2011-01-01

    The present invention provides proteins/genes, which are essential for survival, and consequently, for virulence of Streptococcus pneumoniae in vivo, and thus are ideal vaccine candidates for a vaccine preparation against pneumococcal infection. Further, also antibodies against said protein(s) are i

  5. NEW VIRULENCE FACTORS OF STREPTOCOCCUS PNEUMONIAE.

    NARCIS (Netherlands)

    Bootsma, Jeanette Hester; Burghout, Pieter Jan; Hermans, Peter Wilhelmus Maria; Bijlsma, Johanna; Kuipers, Oscar; Kloosterman, Tomas Gerrit

    2012-01-01

    The present invention provides proteins/genes, which are essential for survival, and consequently, for virulence of Streptococcus pneumoniae in vivo, and thus are ideal vaccine candidates for a vaccine preparation against pneumococcal infection. Further, also antibodies against said protein(s) are i

  6. Radiology of bacterial pneumonia

    Energy Technology Data Exchange (ETDEWEB)

    Vilar, Jose E-mail: vilar_jlu@gva.es; Domingo, Maria Luisa; Soto, Cristina; Cogollos, Jonathan

    2004-08-01

    Bacterial pneumonia is commonly encountered in clinical practice. Radiology plays a prominent role in the evaluation of pneumonia. Chest radiography is the most commonly used imaging tool in pneumonias due to its availability and excellent cost benefit ratio. CT should be used in unresolved cases or when complications of pneumonia are suspected. The main applications of radiology in pneumonia are oriented to detection, characterisation and follow-up, especially regarding complications. The classical classification of pneumonias into lobar and bronchial pneumonia has been abandoned for a more clinical classification. Thus, bacterial pneumonias are typified into three main groups: Community acquired pneumonia (CAD), Aspiration pneumonia and Nosocomial pneumonia (NP).The usual pattern of CAD is that of the previously called lobar pneumonia; an air-space consolidation limited to one lobe or segment. Nevertheless, the radiographic patterns of CAD may be variable and are often related to the causative agent. Aspiration pneumonia generally involves the lower lobes with bilateral multicentric opacities. Nosocomial Pneumonia (NP) occurs in hospitalised patients. The importance of NP is related to its high mortality and, thus, the need to obtain a prompt diagnosis. The role of imaging in NP is limited but decisive. The most valuable information is when the chest radiographs are negative and rule out pneumonia. The radiographic patterns of NP are very variable, most commonly showing diffuse multifocal involvement and pleural effusion. Imaging plays also an important role in the detection and evaluation of complications of bacterial pneumonias. In many of these cases, especially in hospitalised patients, chest CT must be obtained in order to better depict these associate findings.

  7. Effects of Microgravity on Streptoccoccus Pneumonia

    Science.gov (United States)

    2003-01-01

    These gels were obtained by two-dimensional (2D) electrophoresis, in which proteins move different substances through a polyacrylamide gel matrix based on their molecular weight and total charge in an electric field. The gels illustrate principal investigator David Niesel's findings that exposure to modeled microgravity results in some Streptoccoccus Pneumonia's proteins being upregulated and others being downregulated. In 2D protein profiles of whole cell lysates of Streptoccoccus Pneumonia, 6,304 cultured under normal gravity (left), appear to be expressed at higher levels indicated with black circles. Red circles (right) indicate proteins that were grown under modeled microgravity in a high aspect ratio vessel HARV).

  8. Pneumonia (For Parents)

    Science.gov (United States)

    ... Old Feeding Your 1- to 2-Year-Old Pneumonia KidsHealth > For Parents > Pneumonia A A A What's in this article? Signs ... Doctor Professional Treatment Home Care en español Neumonía Pneumonia is a general term for lung infections that ...

  9. Pneumocystis Pneumonia (For Parents)

    Science.gov (United States)

    ... Feeding Your 1- to 2-Year-Old Pneumocystis Pneumonia KidsHealth > For Parents > Pneumocystis Pneumonia Print A A A What's in this article? About PCP Diagnosing PCP Treating PCP Pneumocystis pneumonia (PCP) is an infection caused by Pneumocystis jiroveci , ...

  10. Transcriptional Analysis of 10 Selected Genes in a Model of Penicillin G Induced Persistence of Chlamydophila psittaci in HeLa Cells.

    Science.gov (United States)

    Hu, Yanqun; Chen, Lili; Wang, Chuan; Xie, Yafeng; Chen, Zhixi; Liu, Liangzhuan; Su, Zehong; Wu, Yimou

    2015-08-01

    Chlamydophila psittaci is an important intracellular pathogen. Persistent infection is an important state of the host-parasite interaction in this chlamydial infection, which plays a significant role in spreading the organism within animal populations and in causing chronic chlamydiosis and serious sequelae. In this study, a C. psittaci persistent infection cell model was induced by penicillin G, and real-time quantitative PCR was used to study the transcriptional levels of 10 C. psittaci genes (dnaA, dnaK, ftsW, ftsY, grpE, rpsD, incC, omcB, CPSIT_0846, and CPSIT_0042) in acute and penicillin-G-induced persistent infection cultures. Compared with the acute cultures, the penicillin-G-treated cultures showed a reduced chlamydial inclusion size and a significantly decreased number of elementary body particles. Additionally, some enlarged aberrant reticulate body particles were present in the penicillin- G-treated cultures but not the acute ones. The expression levels of genes encoding products for cell division (FtsW, FtsY) and outer membrane protein E encoding gene (CPSIT_0042) were downregulated (p 0.05). These results provide new insight regarding molecular activities that accompany persistence of C. psittaci, which may play important roles in the pathogenesis of C. psittaci infection.

  11. Diversity of Pneumolysin and Pneumococcal Histidine Triad Protein D of Streptococcus pneumoniae Isolated from Invasive Diseases in Korean Children.

    Science.gov (United States)

    Yun, Ki Wook; Lee, Hyunju; Choi, Eun Hwa; Lee, Hoan Jong

    2015-01-01

    Pneumolysin (Ply) and pneumococcal histidine triad protein D (PhtD) are candidate proteins for a next-generation pneumococcal vaccine. We aimed to analyze the genetic diversity and antigenic heterogeneity of Ply and PhtD for 173 pneumococci isolated from invasive diseases in Korean children. Allele was designated based on the variation of amino acid sequence. Antigenicity was predicted by the amino acid hydrophobicity of the region. There were seven and 39 allele types for the ply and phtD genes, respectively. The nucleotide sequence identity was 97.2%-99.9% for ply and 91.4%-98.0% for phtD gene. Only minor variations in hydrophobicity were noted among the antigenicity plots of Ply and PhtD. Overall, the allele types of the ply and phtD genes were remarkably homogeneous, and the antigenic diversity of the corresponding proteins was very limited. The Ply and PhtD could be useful antigens for universal pneumococcal vaccines.

  12. Clinical characteristics of Q fever and etiology of community-acquired pneumonia in a tropical region of southern Taiwan: a prospective observational study.

    Directory of Open Access Journals (Sweden)

    Chung-Hsu Lai

    Full Text Available The clinical characteristics of Q fever are poorly identified in the tropics. Fever with pneumonia or hepatitis are the dominant presentations of acute Q fever, which exhibits geographic variability. In southern Taiwan, which is located in a tropical region, the role of Q fever in community-acquired pneumonia (CAP has never been investigated.During the study period, May 2012 to April 2013, 166 cases of adult CAP and 15 cases of acute Q fever were prospectively investigated. Cultures of clinical specimens, urine antigen tests for Streptococcus pneumoniae and Legionella pneumophila, and paired serologic assessments for Mycoplasma pneumoniae, Chlamydophila pneumoniae, and Q fever (Coxiella burnetii were used for identifying pathogens associated with CAP. From April 2004 to April 2013 (the pre-study period, 122 cases of acute Q fever were also included retrospectively for analysis. The geographic distribution of Q fever and CAP cases was similar. Q fever cases were identified in warmer seasons and younger ages than CAP. Based on multivariate analysis, male gender, chills, thrombocytopenia, and elevated liver enzymes were independent characteristics associated with Q fever. In patients with Q fever, 95% and 13.5% of cases presented with hepatitis and pneumonia, respectively. Twelve (7.2% cases of CAP were seropositive for C. burnetii antibodies, but none of them had acute Q fever. Among CAP cases, 22.9% had a CURB-65 score ≧2, and 45.8% had identifiable pathogens. Haemophilus parainfluenzae (14.5%, S. pneumoniae (6.6%, Pseudomonas aeruginosa (4.8%, and Klebsiella pneumoniae (3.0% were the most common pathogens identified by cultures or urine antigen tests. Moreover, M. pneumoniae, C. pneumoniae, and co-infection with 2 pathogens accounted for 9.0%, 7.8%, and 1.8%, respectively.In southern Taiwan, Q fever is an endemic disease with hepatitis as the major presentation and is not a common etiology of CAP.

  13. Polymorphic membrane protein (PMP) 20 and PMP 21 of Chlamydia pneumoniae induce proinflammatory mediators in human endothelial cells in vitro by activation of the nuclear factor-kappaB pathway.

    Science.gov (United States)

    Niessner, Alexander; Kaun, Christoph; Zorn, Gerlinde; Speidl, Walter; Türel, Zeynep; Christiansen, Gunna; Pedersen, Anna-Sofie; Birkelund, Svend; Simon, Susan; Georgopoulos, Apostolos; Graninger, Wolfgang; de Martin, Rainer; Lipp, Joachim; Binder, Bernd R; Maurer, Gerald; Huber, Kurt; Wojta, Johann

    2003-07-01

    We tested whether polymorphic membrane proteins (PMPs) of Chlamydia pneumoniae might play a role in triggering an inflammatory response in human endothelial cells. Of 15 purified, recombinant chlamydial PMPs tested, 2 (PMP 20 and PMP 21) dose-dependently increased the production of the inflammatory mediators interleukin (IL)-6 and monocyte chemoattractant protein-1 (MCP-1), in cultured human endothelial cells; production of IL-8 was also increased. When endothelial cells were infected by live C. pneumoniae, an increase in the production of IL-6, IL-8, and MCP-1 was seen. We used adenovirus-induced overexpression of IkappaBalpha-an inhibitor of nuclear factor (NF)-kappaB-to demonstrate that PMP 20 and PMP 21 increase the production of IL-6 and MCP-1 in human endothelial cells by activation of the NF-kappaB pathway, because, in cells overexpressing IkappaBalpha, treatment with the respective PMP did not result in increased production of IL-6 and MCP-1. Thus, C. pneumoniae could, by interactions of its PMPs with the endothelium, contribute to the process of vascular injury during the development and progression of atherosclerotic lesions.

  14. Role of Serum Mycoplasma pneumoniae IgA, IgM, and IgG in the Diagnosis of Mycoplasma pneumoniae-Related Pneumonia in School-Age Children and Adolescents.

    Science.gov (United States)

    Lee, Wei-Ju; Huang, Eng-Yen; Tsai, Chih-Min; Kuo, Kuang-Che; Huang, Yi-Chuan; Hsieh, Kai-Sheng; Niu, Chen-Kuang; Yu, Hong-Ren

    2017-01-01

    Mycoplasma pneumoniae is an important causative pathogen of community-acquired pneumonia in children. Rapid and reliable laboratory diagnosis of M. pneumoniae infection is important so that appropriate antibiotic treatment can be initiated to reduce the misuse of drugs and resistance rates. Anti-M. pneumoniae immunoglobulin M (IgM) is an indicator of recent primary infection but can persist for several months after initial infection. It has been suggested that anti-M. pneumoniae immunoglobulin A (IgA) can be a reliable indicator for recent M. pneumoniae infection in adults. We investigated the clinical diagnostic value of M. pneumoniae IgA in school-age children and adolescents with M. pneumoniae-related pneumonia. Eighty children with pneumonia and seropositive for M. pneumoniae IgM or with a 4-fold increase of anti-M. pneumoniae immunoglobulin G (IgG) were enrolled from May 2015 to March 2016. The titers of M. pneumoniae IgA, IgM, and IgG, the clinical features, and laboratory examinations of blood, C-reactive protein, and liver enzymes were analyzed. The initial positivity rates for M. pneumoniae IgM and IgA upon admission to the hospital were 63.6 and 33.8%, respectively. One week after admission, the cumulative positivity rates for M. pneumoniae IgM and IgA increased to 97.5 and 56.3%, respectively. Detection of M. pneumoniae IgM was more sensitive than detection of M. pneumoniae IgA for the diagnosis of M. pneumoniae-related pneumonia in school-age children and adolescents; however, paired sera are necessary for a more accurate diagnosis.

  15. A novel initiation mechanism of death in Streptococcus pneumoniae induced by the human milk protein-lipid complex HAMLET and activated during physiological death.

    Science.gov (United States)

    Clementi, Emily A; Marks, Laura R; Duffey, Michael E; Hakansson, Anders P

    2012-08-03

    To cause colonization or infection, most bacteria grow in biofilms where differentiation and death of subpopulations is critical for optimal survival of the whole population. However, little is known about initiation of bacterial death under physiological conditions. Membrane depolarization has been suggested, but never shown to be involved, due to the difficulty of performing such studies in bacteria and the paucity of information that exists regarding ion transport mechanisms in prokaryotes. In this study, we performed the first extensive investigation of ion transport and membrane depolarization in a bacterial system. We found that HAMLET, a human milk protein-lipid complex, kills Streptococcus pneumoniae (the pneumococcus) in a manner that shares features with activation of physiological death from starvation. Addition of HAMLET to pneumococci dissipated membrane polarity, but depolarization per se was not enough to trigger death. Rather, both HAMLET- and starvation-induced death of pneumococci specifically required a sodium-dependent calcium influx, as shown using calcium and sodium transport inhibitors. This mechanism was verified under low sodium conditions, and in the presence of ionomycin or monensin, which enhanced pneumococcal sensitivity to HAMLET- and starvation-induced death. Pneumococcal death was also inhibited by kinase inhibitors, and indicated the involvement of Ser/Thr kinases in these processes. The importance of this activation mechanism was made evident, as dysregulation and manipulation of physiological death was detrimental to biofilm formation, a hallmark of bacterial colonization. Overall, our findings provide novel information on the role of ion transport during bacterial death, with the potential to uncover future antimicrobial targets.

  16. A Novel Initiation Mechanism of Death in Streptococcus pneumoniae Induced by the Human Milk Protein-Lipid Complex HAMLET and Activated during Physiological Death*

    Science.gov (United States)

    Clementi, Emily A.; Marks, Laura R.; Duffey, Michael E.; Hakansson, Anders P.

    2012-01-01

    To cause colonization or infection, most bacteria grow in biofilms where differentiation and death of subpopulations is critical for optimal survival of the whole population. However, little is known about initiation of bacterial death under physiological conditions. Membrane depolarization has been suggested, but never shown to be involved, due to the difficulty of performing such studies in bacteria and the paucity of information that exists regarding ion transport mechanisms in prokaryotes. In this study, we performed the first extensive investigation of ion transport and membrane depolarization in a bacterial system. We found that HAMLET, a human milk protein-lipid complex, kills Streptococcus pneumoniae (the pneumococcus) in a manner that shares features with activation of physiological death from starvation. Addition of HAMLET to pneumococci dissipated membrane polarity, but depolarization per se was not enough to trigger death. Rather, both HAMLET- and starvation-induced death of pneumococci specifically required a sodium-dependent calcium influx, as shown using calcium and sodium transport inhibitors. This mechanism was verified under low sodium conditions, and in the presence of ionomycin or monensin, which enhanced pneumococcal sensitivity to HAMLET- and starvation-induced death. Pneumococcal death was also inhibited by kinase inhibitors, and indicated the involvement of Ser/Thr kinases in these processes. The importance of this activation mechanism was made evident, as dysregulation and manipulation of physiological death was detrimental to biofilm formation, a hallmark of bacterial colonization. Overall, our findings provide novel information on the role of ion transport during bacterial death, with the potential to uncover future antimicrobial targets. PMID:22700972

  17. Chlamydia pneumoniae and Atherosclerosis: The End?

    Directory of Open Access Journals (Sweden)

    LE Nicolle

    2005-01-01

    Full Text Available In this issue of the Journal, Patrick et al (pages 298-300 report on the results of a pilot study testing the hypothesis that seropositivity to Chlamydia pneumoniae together with a specific bacteriophage protein is associated with first-episode myocardial infarction or unstable angina. The study evolved from an earlier report suggesting that C pneumoniae with phage seropositivity was strongly associated with the presence of abdominal aortic aneurysm. The phage association suggested a potential explanation for some of the variability in previous studies exploring C pneumoniae as a cause for atherosclerosis (ie, only selected strains of C pneumoniae were pathogenic. Patrick et al found no significant association or trend, and the authors concluded that the negative findings in their pilot study did not support further studies to address this potential association.

  18. Penicillin-Binding Protein Transpeptidase Signatures for Tracking and Predicting β-Lactam Resistance Levels in Streptococcus pneumoniae

    Directory of Open Access Journals (Sweden)

    Yuan Li

    2016-06-01

    Full Text Available β-Lactam antibiotics are the drugs of choice to treat pneumococcal infections. The spread of β-lactam-resistant pneumococci is a major concern in choosing an effective therapy for patients. Systematically tracking β-lactam resistance could benefit disease surveillance. Here we developed a classification system in which a pneumococcal isolate is assigned to a “PBP type” based on sequence signatures in the transpeptidase domains (TPDs of the three critical penicillin-binding proteins (PBPs, PBP1a, PBP2b, and PBP2x. We identified 307 unique PBP types from 2,528 invasive pneumococcal isolates, which had known MICs to six β-lactams based on broth microdilution. We found that increased β-lactam MICs strongly correlated with PBP types containing divergent TPD sequences. The PBP type explained 94 to 99% of variation in MICs both before and after accounting for genomic backgrounds defined by multilocus sequence typing, indicating that genomic backgrounds made little independent contribution to β-lactam MICs at the population level. We further developed and evaluated predictive models of MICs based on PBP type. Compared to microdilution MICs, MICs predicted by PBP type showed essential agreement (MICs agree within 1 dilution of >98%, category agreement (interpretive results agree of >94%, a major discrepancy (sensitive isolate predicted as resistant rate of <3%, and a very major discrepancy (resistant isolate predicted as sensitive rate of <2% for all six β-lactams. Thus, the PBP transpeptidase signatures are robust indicators of MICs to different β-lactam antibiotics in clinical pneumococcal isolates and serve as an accurate alternative to phenotypic susceptibility testing.

  19. A novel method for rapid detection of Streptococcus pneumoniae antigens in blood.

    Science.gov (United States)

    Fukushima, Kiyoyasu; Kubo, Toru; Ehara, Naomi; Nakano, Reiji; Matsutake, Toyoshi; Ishimatu, Yuji; Tanaka, Yumi; Akamatsu, Suguru; Izumikawa, Koichi; Kohno, Shigeru

    2016-03-01

    In this study, we used "RAPIRUN(®)Streptococcus pneumoniae HS (otitis media/sinusitis) (RAPIRUN-HS)," a rapid S. pneumoniae antigen detection kit, to investigate methods for detecting S. pneumoniae antigens in blood of 32 bacterial pneumonia patients. We simultaneously performed PCR to detect S. pneumoniae in blood samples. The results of these tests were compared based on pneumonia severity, determined using the Pneumonia Severity Index (PSI) score classification. Four S. pneumoniae PCR-positive patients of the six severe pneumococcal pneumonia patients (PSI risk class IV/V) also tested positive using RAPIRUN-HS. Twenty-four mild to moderate pneumonia patients (PSI risk class I-III) were S. pneumoniae PCR-negative; of these, 21 tested negative using RAPIRUN-HS. The pneumococcal pneumonia patients testing positive using RAPIRUN-HS had low leukocyte counts and elevated C-reactive protein and procalcitonin levels, indicating that RAPIRUN-HS results were correlated with pneumonia severity. The time course evaluations of the laboratory tests for severe pneumococcal pneumonia patients showed that RAPIRUN-HS and S. pneumoniae PCR yielded positive results earlier than the changes in procalcitonin and IL-6. Thus, concomitant pneumococcal bacteremia was strongly suspected in patients testing positive using RAPIRUN-HS. In conclusion, RAPIRUN-HS may be useful for determining whether to admit patients into hospitals and selecting the appropriate antimicrobial agents.

  20. Psychosis following mycoplasma pneumonia.

    Science.gov (United States)

    Banerjee, Bonita; Petersen, Kyle

    2009-09-01

    Extrapulmonary manifestations of Mycoplasma pneumoniae are well described, including a subset of central nervous system (CNS)-associated syndromes. In pediatric populations, frequencies of CNS sequelae occur in 0.1% to 7% of patients. Neurologic illness associated with M. pneumoniae, such as meningitis, encephalitis, polyradiculitis, Guillain-Barre, and stroke have been reported; however, the incidence of M. pneumoniae-associated organic brain syndrome is rare. We present the case of a 20-year-old midshipman with acute psychosis following resolution of M. pneumoniae pneumonia and review 6 other adult cases found in the literature. M. pneumoniae remains one of the most common causes of respiratory illnesses in the military recruit setting and therefore should always be suspected as an organic cause of mental status changes in young persons such as recruits, cadets, and midshipmen particularly with antecedent respiratory illnesses.

  1. Bactericidal activities of parenteral antibiotics and genotype of penicillin-binding protein in Streptococcus pneumoniae and Haemophilus influenzae isolated from children's blood.

    Science.gov (United States)

    Sakata, Hiroshi

    2006-10-01

    A total of 16 isolates of Streptococcus pneumoniae and 18 isolates of Haemophilus influenzae were obtained from the blood of children admitted to the pediatric wards of hospitals in Hokkaido Kamikawa subprefecture between January 2003 and December 2005. The ages of the patients with S. pneumoniae or H. influenzae infection ranged from 2 months to 9 years and from 1 month to 4 years, respectively. The diagnoses of S. pneumoniae infection were as follows: pneumonia in 8 patients, occult bacteremia in 5 patients, and meningitis in 3 patients. The diagnoses of H. influenzae were: meningitis in 6 patients, pneumonia in 4 patients, occult bacteremia in 4 patients, epiglotitis in 2 patients, and facial cellulitis in 2 patients. Out of 16 S. pneumoniae isolates, penicillin-resistant strains with a mutation of 3 genes were observed in 7 children, and penicillin intermediate-resistant strains with a mutation of 1 or 2 genes were observed in 8 children. Out of 18 H. influenzae isolates, the beta-lactamase-negative ampicillin-resistant strain with a substitution of 2 points in the ftsI gene was revealed in 2 children, the beta-lactamase-negative ampicillin-resistant strain with a substitution of 1 point in the ftsI gene was observed in 4 children, the beta-lactamase-positive amoxicillin/clavulanic acid-resistant strain with blaTEM-1 and ftsI with 2 substitutions in the ftsI gene was observed in 3 children, and the beta-lactamase-positive ampicillin-resistant strain with blaTEM-1was not observed. The MBC90s of ampicillin, ceftriaxone, cefotaxime, meropenem, panipenem, and vancomycin against S. pneumoniae were 8 microg/ml, 1 microg/ml, 1 microg/ml 1 microg/ml, 0.25 microg/ml, and 0.5 microg/ml, respectively. Those of ampicillin, piperacillin, ceftriaxone, cefotaxime, meropenem, and panipenem against H. influenzae were >128 microg/ml, >128 microg/ml, 0.25 microg/mL, 1 microg/ml, 0.12 microg/ml, and 0.5 g/ml, respectively. It is suggest that the minimum bactricidal concentration

  2. Cytotoxicity of Mycoplasma pneumoniae Membranes

    Science.gov (United States)

    Gabridge, Michael G.; Johnson, Cynthia K.; Cameron, Alexander M.

    1974-01-01

    Organ cultures of adult hamster trachea were used to evaluate the cytotoxic potential of cell fractions of Mycoplasma pneumoniae. Cytoplasm was essentially devoid of activity, whereas viable cells and membrane preparations, at a level of 25 μg of protein per ml, induced necrosis. Damage, as revealed by light and electron microscopy, included ciliostasis, vacuolization, loss of ciliated respiratory epithelial cells, disorganization, and a loss of polarity. Dose response data indicated that the speed and degree of cytotoxicity was directly related to the concentration of membranes. Doses of 30 to 60 μg of protein per ml could reduce relative ciliary activity to 20% of the control level within 4 days. Membranes prepared after freeze-thaw lysis of cells were almost twice as active as those isolated after a combination of osmotic and sonic shock. Membranes of M. fermentans were inactive, though both the FH and M129 strains of M. pneumoniae were toxic. These data indicate that the toxic factor responsible for M. pneumoniae may be located in the cell membrane. Images PMID:16558100

  3. Detection and identification of Chlamydophila psittaci in asymptomatic parrots in Poland

    Directory of Open Access Journals (Sweden)

    Piasecki Tomasz

    2012-12-01

    Full Text Available Abstract Background Psittacosis, an avian disease caused by Chlamydophila psittaci, can manifest as an acute, protracted, or chronic illness, but can also be asymptomatic. C. psittaci can persist in the host for months to years, often without causing obvious illness, and therefore poses a threat for zoonotic outbreak. We investigated the prevalence of C. psittaci from 156 tracheal swab samples from 34 different species of parrots in Poland, and determined the genotype of strains from the positive samples. Results An overall prevalence of 10.3% was observed using two different PCR assays, both providing similar results. Thirteen of the PCR-positive samples were genotype A, two were genotype B, and one could not be classified. Conclusions These results indicate widespread dissemination of C. psittaci in Polish psittacine populations, without any clinical signs of chlamydiosis, and hence could pose a zoonotic hazard. PCR screening provided a definitive diagnosis of psittacosis, and subsequent ompA gene analysis could be helpful for better understanding the epidemiology of the C. psittaci genotypes. To the best of our knowledge, this is the first report of the incidence of C. psittaci in parrots in Poland.

  4. Evaluation of bioaerosol sampling techniques for the detection of Chlamydophila psittaci in contaminated air.

    Science.gov (United States)

    Van Droogenbroeck, Caroline; Van Risseghem, Marleen; Braeckman, Lutgart; Vanrompay, Daisy

    2009-03-16

    Chlamydophila (C.) psittaci, a category B bioterrorism agent, causes respiratory disease in birds and psittacosis or parrot fever in man. The disease spreads aerogenically and no vaccines are available for either birds or man. Highly sensitive C. psittaci bioaerosol monitoring methods are unavailable. We evaluated: (1) dry filtration for collecting C. psittaci from contaminated air using different samplers and membrane filters, (2) impingement into different liquid collection media by use of the AGI-30 impinger and the BioSampler and (3) impaction into newly designed C. psittaci media utilizing the MAS-100 aerosol impactor. For personal bioaerosol sampling, we recommend the use of a gelatin filter in combination with the IOM inhalable dust sampler at an airflow rate of 2L/min. This allowed the detection of 10 organisms of C. psittaci by both PCR and culture. For stationary bioaerosol monitoring, sampling 1000L of air in 10min with the MAS-100 impactor and ChlamyTrap 1 impaction medium was most efficient and made it possible to detect 1 and 10 C. psittaci organisms by PCR and culture, respectively. ChlamyTrap 1 in combination with the MAS-100 impactor might also be applicable for bioaerosol monitoring of viruses.

  5. Ovine trophoblast is a primary source of TNFalpha during Chlamydophila abortus infection.

    Science.gov (United States)

    Wheelhouse, Nick; Wattegedera, Sean; Stanton, James; Maley, Stephen; Watson, Donna; Jepson, Catherine; Deane, David; Buxton, David; Longbottom, David; Baszler, Tim; Entrican, Gary

    2009-06-01

    Chlamydophila abortus is a Gram-negative obligate intracellular bacterium that causes infectious abortion in sheep (ovine enzootic abortion, OEA) and humans. Infected placentas recovered from sheep that experience OEA have thickened membranes, contain dense inflammatory cellular infiltrates and show evidence of intravascular thrombosis. Despite widespread inflammation, chlamydial multiplication is restricted to the chorionic trophoblast cells. To investigate the potential role of trophoblast in the initiation and propagation of placental inflammation during OEA, the AH-1 ovine trophoblast cell line was experimentally infected with C. abortus and analysed for the release of pro-inflammatory mediators. C. abortus was found to induce the release of both tumour necrosis factor-alpha (TNFalpha) and CXCL8 (interleukin-8) from AH-1 cells in a dose- and time-dependent manner. Ultra-violet (UV)-killed organisms did not elicit this profile, indicating that intracellular multiplication of C. abortus was required for release of these pro-inflammatory mediators. Exposure of AH-1 cells to recombinant ovine TNFalpha alone resulted in the release of CXCL8, suggestive of a self-propagating inflammatory cytokine and chemokine cascade. These data indicate a primary role for trophoblast in the initiation and propagation of placental inflammation during chlamydial abortion.

  6. Molecular detection of Chlamydophila abortus in post-abortion sheep at oestrus and subsequent lambing.

    Science.gov (United States)

    Livingstone, Morag; Wheelhouse, Nicholas; Maley, Stephen W; Longbottom, David

    2009-03-16

    Enzootic abortion of ewes (EAE), resulting from infection with the bacterium Chlamydophila abortus (C. abortus), is a major cause of lamb loss in Europe. The purpose of this study was to assess the potential impact of the shedding of organisms in post-abortion ewes at oestrus and subsequent lambing on the epidemiology of EAE. Using a newly developed C. abortus specific real-time PCR assay, few chlamydial genomes could be detected in vaginal swabs taken from post-abortion ewes at oestrus. At subsequent parturition, all ewes lambed normally with no macroscopic or microbiological evidence of infection. Real-time PCR analysis of placental samples identified very few or no chlamydial genomes, which contrasted significantly with samples taken at the time of abortion, where an average of 2.7x10(7) chlamydial genomes per microgram of total tissue DNA was detected. Few genomes could also be detected from vaginal and cervical tissue samples and lymph nodes taken post-mortem. The results, although not discounting the possibility of a chronic low level persistent infection in post-abortion ewes, suggest that the low levels of chlamydial DNA detected during the periovulation period and at lambing do not significantly impact on the epidemiology of EAE. In terms of flock management, the products of abortion should be considered the major and principal source of infection for transmission to naïve ewes.

  7. Epidemiology of Chlamydophila caviae-like Chlamydia Isolated from Urethra and Uterine Cervix

    Directory of Open Access Journals (Sweden)

    Kariyama,Reiko

    2010-02-01

    Full Text Available In 2000, chlamydial strains OK133 and OK135 were isolated from 2 female patients with cervicitis. These strains were unresponsive to commercially available PCR and LCR test kits for the diagnosis of Chlamydia trachomatis infection, and their phenotypic characteristics were very similar. The OK135 nucleotide sequence in MOMP-VD2 gene closely resembled that of Chlamydophila caviae GPIC. A similar strain was isolated in 2003 from a male patient OKM2 with urethritis, from which the strain SC10-6 was cloned by the plaque purification method. The nucleotide sequence of the entire MOMP gene of SC10-6 was exactly the same as that of OK135. Thus, the strains OK135 and SC10-6, together with OK133, have been called C. caviae-like Chlamydia. We designed primers for nested PCR assay, the product of which showed a single-band 311-bp fragment, to detect C. caviae-like Chlamydia. Of swab specimens obtained from 202 patients from 2003 to 2006 (119 male and 83 female patients, 18 specimens (8.9% from 14 male and 4 female patients were positive, suggesting that C. caviae-like Chlamydia infection is rather common. Thus far, it has not been determined whether C. caviae-like Chlamydia is pathogenic for humans.

  8. Prevalence of antibodies against Chlamydophila abortus and Coxiella burnetii in goat herds in Poland.

    Science.gov (United States)

    Czopowicz, M; Kaba, J; Szaluś-Jordanow, O; Nowicki, M; Witkowski, L; Nowicka, D; Frymus, T

    2010-01-01

    An epidemiological study was carried out to determine the herd prevalence of Chlamydophila abortus and Coxiella burnetii antibodies in goats covered by a milk recording program in Poland. The survey took place in 2007 and 48 herds located in different parts of the country were involved. A representative sample from each herd was taken by a simple random sampling allowing to detect seropositivity of a herd on a 95% level of confidence. In total 918 goats were tested for specific antibodies against both germs with the use of enzyme-linked immunosorbent assays. In addition, history of reproductive failures was recorded in these herds. The survey revealed that the herd prevalence of C. abortus was 4.2% (2 herds) while no C. burnetii antibodies were found. Abortions were reported to be a problem in 80% of herds while repeating estrus was encountered in 46% of herds. Reproductive failure concerned two seropositive herds as well. Since the germ is present in the population, it has to be taken into consideration in diagnostic process. Nevertheless, the results of the present study indicate that C. abortus infection occurs infrequently in Polish goats. As no antibodies against C. burnetii were detected in the screened sample the risk of goat-to-human transmission of both bacteria in Poland seems to be very low.

  9. Inflammatory cytokine responses in a pregnant mouse model of Chlamydophila abortus infection.

    Science.gov (United States)

    Kerr, Karen; Wheelhouse, Nicholas; Livingstone, Morag; Anderson, Ian E; Entrican, Gary; McKeever, Declan; Longbottom, David

    2010-08-26

    Chlamydophila abortus (C. abortus) is the aetiological agent of ovine enzootic abortion (OEA). The highly elevated expression of the pro-inflammatory cytokine tumour necrosis factor-alpha (TNFalpha) and low-level expression of interferon-gamma (IFNgamma) that are detected in C. abortus-infected placentas have been implicated in the pathogenesis of OEA. Late-term abortions similar to those occurring in sheep have also been observed in mouse models of C. abortus infection. Since mouse studies have contributed significantly to our understanding of the immunological responses to chlamydial infections and serve as a good model for rapidly assessing candidate vaccines for OEA, we investigated local expression of TNFalpha and IFNgamma in infected mice. At various time points over the course of infection mice were sacrificed, serum samples obtained for serum antibody and cytokine analyses, and livers and placental tissues were removed and fixed to determine C. abortus colonisation and cytokine expression. Immunostaining for C. abortus was significantly greater in placenta compared to liver (P<0.001), whereas local IFNgamma expression was lower and TNFalpha expression was absent in the placenta compared with the liver across all time points. Serum concentrations of both IFNgamma and TNFalpha increased throughout pregnancy in infected mice. These data suggest that a protective systemic inflammatory immune response controls maternal C. abortus infection but not placental/fetal infection in mice. In contrast to sheep, murine placental TNFalpha expression does not correlate with C. abortus infection, suggesting that the immunopathogenesis of chlamydial abortion differs in these species.

  10. Epidemiology of Chlamydophila caviae-like Chlamydia isolated from urethra and uterine cervix.

    Science.gov (United States)

    Murao, Wataru; Wada, Koichiro; Matsumoto, Akira; Fujiwara, Michihisa; Fukushi, Hideto; Kishimoto, Toshio; Monden, Koichi; Kariyama, Reiko; Kumon, Hiromi

    2010-02-01

    In 2000, chlamydial strains OK133 and OK135 were isolated from 2 female patients with cervicitis. These strains were unresponsive to commercially available PCR and LCR test kits for the diagnosis of Chlamydia trachomatis infection, and their phenotypic characteristics were very similar. The OK135 nucleotide sequence in MOMP-VD2 gene closely resembled that of Chlamydophila caviae GPIC. A similar strain was isolated in 2003 from a male patient OKM2 with urethritis, from which the strain SC10-6 was cloned by the plaque purification method. The nucleotide sequence of the entire MOMP gene of SC10-6 was exactly the same as that of OK135. Thus, the strains OK135 and SC10-6, together with OK133, have been called C. caviae-like Chlamydia. We designed primers for nested PCR assay, the product of which showed a single-band 311-bp fragment, to detect C. caviae-like Chlamydia. Of swab specimens obtained from 202 patients from 2003 to 2006 (119 male and 83 female patients), 18 specimens (8.9%) from 14 male and 4 female patients were positive, suggesting that C. caviae-like Chlamydia infection is rather common. Thus far, it has not been determined whether C. caviae-like Chlamydia is pathogenic for humans.

  11. Seroprevalence of Chlamydophila abortus infection in yaks (Bos grunniens) in Qinghai, China.

    Science.gov (United States)

    Chen, Qiwei; Gong, Xiaowei; Zheng, Fuying; Cao, Xiaoan; Li, Zhaocai; Zhou, Jizhang

    2014-03-01

    Chlamydophila abortus is an important amphixenosis which in a wide range of animals, associated with reproductive disorders in yaks. In order to assess the prevalence of this infection in yaks in Qinghai, China, a cross-sectional study was carried out, and a total of 674 serum samples were collected from June to October 2012 in six counties, and antibodies to C. abortus were examined by indirect hemagglutination (IHA) test. The overall seroprevalence of C. abortus in yaks was 17.66 % (119/674), and the seroprevalence of antibodies to C. abortus in yaks ranged from 11.82 to 28.43 % among the six different areas, and the difference was statistically significant (P  0.05). The seroprevalence of C. abortus infection in male yak (16.8 %) was slightly lower than that in females (17.85 %), and the difference was not statistically significant (P > 0.05). So far, this is the first systematic and comprehensive investigation of C. abortus infectionin in yaks in this area.

  12. Evidence of Chlamydophila abortus vaccine strain 1B as a possible cause of ovine enzootic abortion.

    Science.gov (United States)

    Wheelhouse, Nicholas; Aitchison, Kevin; Laroucau, Karine; Thomson, Jill; Longbottom, David

    2010-08-09

    Chlamydophila abortus, the agent of ovine enzootic abortion (OEA), is a major cause of lamb mortality worldwide. Disease can be controlled through the use of vaccines based on the 1B temperature-sensitive mutant strain of C. abortus. This study investigated suspected OEA cases across Scotland for the presence of the 1B strain by analysis of recently identified unique point mutations (9). Thirty-five cases were C. abortus-positive and 14 came from vaccinated flocks. Analysis of single nucleotide polymorphisms by PCR-RFLP and sequence analysis revealed the presence of point mutations consistent with the presence of the 1B vaccine strain in 5 of these 14 samples. Quantitative real-time PCR revealed comparable numbers of genome copies of the 1B strain in infected placentas to those present following wild-type infection. This study is the first to demonstrate the presence of the 1B vaccine strain in the placentas of OEA cases and suggests a probable causal role in the disease.

  13. Ocorrência de Chlamydophila psittaci em pombos (Columba livia na cidade de Salvador, Bahia

    Directory of Open Access Journals (Sweden)

    D.C. Leal

    2015-06-01

    Full Text Available A existência de populações numerosas de pombos (Columba livia em centros urbanos, em quase todo o mundo, tem se tornado um risco à saúde pública em vista das zoonoses que podem transmitir. A infecção por Chlamydophila psittaci foi avaliada em pombos que frequentam áreas públicas, como praças, ruas e pontos turísticos na cidade de Salvador, Bahia, por meio da PCR em amostras de fezes frescas, suabes cloacais e orofaríngeos. O estudo revelou uma frequência de infecção por C. psittaci de 11,7% (16/137 dos pombos, e oito dos 10 locais pesquisados apresentavam aves infectadas. A detecção de C. psittaci em amostras de cloaca e orofaringe foi maior (15,8% - 3/19 que em amostras de fezes frescas (11% - 13/118. Os resultados demonstram a ocorrência de infecção por C. psittaci em pombos que habitam as áreas públicas da cidade de Salvador e apontam para a necessária elaboração de medidas de controle e monitoramento das populações de pombos urbanos, bem como de ações voltadas à conscientização da sociedade sobre os riscos à saúde pública.

  14. Detection of antibodies against Chlamydophila abortus in Costa Rican sheep flocks

    Directory of Open Access Journals (Sweden)

    R. Villagra-Blanco

    2015-09-01

    Full Text Available A total of 359 sheep samples from 15 flocks were analyzed for the presence of antibodies against Chlamydophila abortus using a commercial Enzyme linked Immunosorbent Assay (ELISA. Antibodies were detected in 19 (5.29% sheep from 12 (80% flocks. Seropositive animals were found in all analyzed regions (Central, Chorotega, Atlantic Huetar, North Huetar and Central Pacific determining prevalence between 0.28% and 4.4%, and intra-flock positivity between 3.7% and 25.0%. The survey revealed two risk factors associated with seropositivity; introducing animals (males and females, embryos, or semen from other farms or from abroad without any sanitary certification, and flocks not having quarantine areas or separated boxes for diseased animals. No clinical signs of disease were observed in positive seroreactors. C. abortus seems to be present in Costa Rica in a very low prevalence in sheep flocks. Further studies, to isolate the bacteria are required. Finally, implementation of control measures to prevent the spread of C. abortus is recommended.

  15. Identification of Chlamydophila abortus and the development of lesions in placental tissues of experimentally infected sheep.

    Science.gov (United States)

    Maley, S W; Livingstone, M; Rodger, S M; Longbottom, D; Buxton, D

    2009-03-16

    Chlamydophila (C.) abortus is a major cause of infectious abortion in sheep in many countries. Twenty-one pregnant sheep were experimentally infected intranasally with C. abortus at 70 days of gestation (dg). Thereafter, a number of animals were killed at weekly intervals and a post-mortem examination was carried out. Evidence of chlamydial infection in the placenta was determined by isolation of the bacterium by tissue culture and detection of C. abortus DNA by real-time polymerase chain reaction (real-time PCR). In addition, histopathological changes in the placenta were assessed, as was the detection of chlamydial antigen by immunohistochemistry (IHC). Evidence of placental infection was observed as early as 2 weeks after inoculation, and while only relatively low numbers of bacteria were isolated by culture and/or detected by real-time PCR prior to 113-114dg, at 119-121dg, it was more numerous. This study, using the four criteria for assessment of infection, showed that while C. abortus gained access to the placenta as early as 85dg, characteristic histopathological changes were not apparent until 119/121dg. While the chronology of when the bacterium arrived in the placenta and subsequent lesion development is remarkable for its consistency this paper provides more reliable data on the former which in turn now allows study of the factors that permit its access to this tissue and govern its multiplication and the ensuing triggering of damage.

  16. Key role of Chlamydophila psittaci on Belgian turkey farms in association with other respiratory pathogens.

    Science.gov (United States)

    Van Loock, M; Geens, T; De Smit, L; Nauwynck, H; Van Empel, P; Naylor, C; Hafez, H M; Goddeeris, B M; Vanrompay, D

    2005-04-25

    Two hundred turkey sera from eight Belgian and two French farms were tested for the presence of antibodies against avian pneumovirus (APV), Ornithobacterium rhinotracheale (ORT), Mycoplasma gallisepticum, Mycoplasma meleagridis and Chlamydophila psittaci. At slaughter, C. psittaci, APV and ORT antibodies were detected in 94, 34 and 6.5% of the turkeys, respectively. No antibodies against M. gallisepticum or M. meleagridis were present. Additionally, turkeys on three Belgian farms were examined from production onset until slaughter using both serology and antigen or gene detection. All farms experienced two C. psittaci infection waves, at 3-6 and 8-12 weeks of age. Each first infection wave was closely followed by an ORT infection starting at the age of 6-8 weeks, which was still detectable when the second C. psittaci infection waves started. Animals on farm A were not vaccinated against APV leading to an APV subtype B outbreak accompanying the first C. psittaci infection wave. Despite subtype A APV vaccination on farms B and C, the second C. psittaci infection waves were accompanied (farm B) or followed (farm C) by a subtype B APV infection. On all farms respiratory signs always appeared together with a proven C. psittaci, APV and/or ORT infection. This study suggests an association between C. psittaci, APV and ORT, and indicates the multi-factorial aetiology of respiratory infections in commercial turkeys. All three pathogens should be considered when developing prevention strategies for respiratory disease.

  17. Update on interstitial pneumonia.

    Science.gov (United States)

    Wilkins, Pamela A; Lascola, Kara M

    2015-04-01

    Interstitial pneumonias encompass a wide variety of acute and chronic respiratory diseases and include the specific diseases equine multinodular pulmonary fibrosis and acute lung injury and acute respiratory distress. These diseases have been diagnosed in all age groups of horses, and numerous agents have been identified as potential causes of interstitial pneumonia. Despite the varied causes, interstitial pneumonia is uniformly recognized by the severity of respiratory disease and often poor clinical outcome. This article reviews the causal agents that have been associated with the development of interstitial pneumonia in horses. Pathophysiology, clinical diagnosis, and treatment options are discussed. Copyright © 2015 Elsevier Inc. All rights reserved.

  18. Klebsiella pneumoniae Siderophores Induce Inflammation, Bacterial Dissemination, and HIF-1α Stabilization during Pneumonia

    Science.gov (United States)

    Holden, Victoria I.; Breen, Paul; Houle, Sébastien; Dozois, Charles M.

    2016-01-01

    ABSTRACT Klebsiella pneumoniae is a Gram-negative pathogen responsible for a wide range of infections, including pneumonia and bacteremia, and is rapidly acquiring antibiotic resistance. K. pneumoniae requires secretion of siderophores, low-molecular-weight, high-affinity iron chelators, for bacterial replication and full virulence. The specific combination of siderophores secreted by K. pneumoniae during infection can impact tissue localization, systemic dissemination, and host survival. However, the effect of these potent iron chelators on the host during infection is unknown. In vitro, siderophores deplete epithelial cell iron, induce cytokine secretion, and activate the master transcription factor hypoxia inducible factor-1α (HIF-1α) protein that controls vascular permeability and inflammatory gene expression. Therefore, we hypothesized that siderophore secretion by K. pneumoniae directly contributes to inflammation and bacterial dissemination during pneumonia. To examine the effects of siderophore secretion independently of bacterial growth, we performed infections with tonB mutants that persist in vivo but are deficient in siderophore import. Using a murine model of pneumonia, we found that siderophore secretion by K. pneumoniae induces the secretion of interleukin-6 (IL-6), CXCL1, and CXCL2, as well as bacterial dissemination to the spleen, compared to siderophore-negative mutants at an equivalent bacterial number. Furthermore, we determined that siderophore-secreting K. pneumoniae stabilized HIF-1α in vivo and that bacterial dissemination to the spleen required alveolar epithelial HIF-1α. Our results indicate that siderophores act directly on the host to induce inflammatory cytokines and bacterial dissemination and that HIF-1α is a susceptibility factor for bacterial invasion during pneumonia. PMID:27624128

  19. Klebsiella pneumoniae Siderophores Induce Inflammation, Bacterial Dissemination, and HIF-1α Stabilization during Pneumonia

    Directory of Open Access Journals (Sweden)

    Victoria I. Holden

    2016-09-01

    Full Text Available Klebsiella pneumoniae is a Gram-negative pathogen responsible for a wide range of infections, including pneumonia and bacteremia, and is rapidly acquiring antibiotic resistance. K. pneumoniae requires secretion of siderophores, low-molecular-weight, high-affinity iron chelators, for bacterial replication and full virulence. The specific combination of siderophores secreted by K. pneumoniae during infection can impact tissue localization, systemic dissemination, and host survival. However, the effect of these potent iron chelators on the host during infection is unknown. In vitro, siderophores deplete epithelial cell iron, induce cytokine secretion, and activate the master transcription factor hypoxia inducible factor-1α (HIF-1α protein that controls vascular permeability and inflammatory gene expression. Therefore, we hypothesized that siderophore secretion by K. pneumoniae directly contributes to inflammation and bacterial dissemination during pneumonia. To examine the effects of siderophore secretion independently of bacterial growth, we performed infections with tonB mutants that persist in vivo but are deficient in siderophore import. Using a murine model of pneumonia, we found that siderophore secretion by K. pneumoniae induces the secretion of interleukin-6 (IL-6, CXCL1, and CXCL2, as well as bacterial dissemination to the spleen, compared to siderophore-negative mutants at an equivalent bacterial number. Furthermore, we determined that siderophore-secreting K. pneumoniae stabilized HIF-1α in vivo and that bacterial dissemination to the spleen required alveolar epithelial HIF-1α. Our results indicate that siderophores act directly on the host to induce inflammatory cytokines and bacterial dissemination and that HIF-1α is a susceptibility factor for bacterial invasion during pneumonia.

  20. Utility of serum ferritin and lactate dehydrogenase as surrogate markers for steroid therapy for Mycoplasma pneumoniae pneumonia.

    Science.gov (United States)

    Kawamata, Ryou; Yokoyama, Koji; Sato, Miori; Goto, Masahide; Nozaki, Yasuyuki; Takagi, Takeshi; Kumagai, Hideki; Yamagata, Takanori

    2015-11-01

    Patients with severe mycoplasma pneumonia having very high serum interleukin-18 levels may require systemic corticosteroid treatment. However, we know of no laboratory markers that have been identified to assess the precise severity of Mycoplasma pneumoniae pneumonia. Thus, we investigated the usefulness of four clinical laboratory tests as severity indicators and surrogate markers for initiation of steroid therapy in these patients. For 22 Japanese children (including 3 patients who needed systemic corticosteroid therapy) diagnosed with Mycoplasma pneumoniae pneumonia, white blood cell counts and serum concentrations of interleukin-18, C-reactive protein, lactate dehydrogenase, and ferritin were determined in the acute and recovery phases. In total, 8 and 14 patients were classified as moderate and mild pneumonia, respectively, according to clinical manifestations. The serum interleukin-18 level in the acute phase of the pneumonia group was significantly higher than that of age-matched controls. Furthermore, serum interleukin-18, lactate dehydrogenase and ferritin levels in the acute phase increased in parallel with the severity of the pneumonia. The serum ferritin level was also higher in the acute phase than in the recovery phase. Positive correlations between the levels of serum interleukin-18, lactate dehydrogenase and ferritin were observed in the acute phase. Serum lactate dehydrogenase and ferritin levels may be useful as indicators of the severity of pediatric Mycoplasma pneumoniae pneumonia for initiation of corticosteroid therapy. Copyright © 2015 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  1. Rhodococcus equi foal pneumonia.

    Science.gov (United States)

    Cohen, Noah D

    2014-12-01

    Pneumonia caused by Rhodococcus equi is an important cause of disease and death in foals. This article reviews current knowledge of the epidemiology, clinical signs, diagnosis, treatment, prevention, and control of R equi pneumonia in foals. Copyright © 2014 Elsevier Inc. All rights reserved.

  2. Bronchitis and Pneumonia

    Science.gov (United States)

    ... by a health care provider. How serious are bronchitis and pneumonia? Both conditions are more serious if a child has a chronic health condition or if the condition is caused by a bacteria, in which case antibiotics are the treatment of choice. When pneumonia is caused by bacteria, ...

  3. Insight into behavior of epithelial cells of the feline conjunctiva in chronic conjunctivitis as a possible limitation in detection of Chlamydophila spp.

    Science.gov (United States)

    Kiełbowicz, Z; Płoneczka-Janeczko, K; Kuropka, P

    2014-01-01

    The aims of this work was documentation of the reactivity of feline conjunctival epithelial cells in chronic conjunctivitis and the investigation of a possible correlation of histological findings in conjunctiva with a limitation in detection of the pathogen. In this observational study, conjunctival swab samples collected from six cats suffering from chronic conjunctivitis were monitored for Chlamydophila spp. infection for one month, every ten days. Chlamydophilosis was diagnosed by conventional PCR, and confirmed by sequencing analysis. A lack of coherence with results in subsequent studies using PCR did not allow an accurate diagnosis. Additional bioptat samples of conjunctiva were collected for diagnostic purposes and stained in haematoxylin and eosin following the Giemsa method for light microscopic analysis. Additionally the samples were incubated for 15 min with IMAGEN Chlamydia conjugate (IMAGEN Chlamydia reagent kit, Dako, UK), allowing immunofluorescence detection of Chlamydophila spp. Within the epithelium an increased number of goblet cells, as well as general enlargement of the epithelium and a reduced number of normal epithelial cells, was observed. Only in areas of low epithelium could structures similar to the elementary bodies of Chlamydophila spp. be distinguished. The presented data document a possible limitation in molecular evidence for chlamydophila infection in some naturally infected cats, taking into account histological conditions in conjunctiva at the same time.

  4. Contract Support and Facilitation of Epidemic Outbreak Surveillance (EOS) Program Final Operating Capability (FOC)

    Science.gov (United States)

    2007-08-01

    Chlamydophila pneumoniae MOMP Gene 232 bp Legionella pneumophila Macrophage Inf. Potentiator 160 bp Legionella micdadei Macrophage Inf. Potentiator 240 bp...developed and successfully demonstrated on the carbon electrode array for a set of pathogens including Mycoplasma pneumoniae, Chlamydophila pneumoniae

  5. Computational studies on the resistance of penicillin-binding protein 2B (PBP2B) of wild-type and mutant strains of Streptococcus pneumoniae against β-lactam antibiotics.

    Science.gov (United States)

    Ramalingam, Jothi; Vennila, Jannet; Subbiah, Parthasarathy

    2013-09-01

    Mutations within transpeptidase domain of penicillin-binding protein 2B of the strains of Streptococcus pneumoniae leads to resistance against β-lactam antibiotics. To uncover the important residues responsible for sensitivity and resistance, the recently determined three dimensional structures of penicillin-binding protein 2B of both wild-type R6 (sensitive) and mutant 5204 (resistant) strains along with the predicted structures of other mutant strains G54, Hungary19A-6 and SP195 were considered for the interaction study with β-lactam antibiotics using induced-fit docking of Schrödinger. Associated binding energies of the complexes and their intermolecular interactions in the binding site clearly show that the wild-type R6 as sensitive, mutant strains 5204 and G54 as highly resistant, and the mutant strains Hungary19A-6 and SP195 as intermediate resistant. The study also reveals that the mutant strains Hungary19A-6 and SP195 exhibit intermediate resistant because of the existence of mutations till the intermediate 538th and 516th positions, respectively, and not till the end of the C-terminus. Furthermore, our investigations show that if the mutations are extended till the end of the C terminus, then the antibiotic resistance of induced-mutated strains increases from intermediate to high as in the strains 5204 and G54. The binding patterns obtained in the study are useful in designing potential inhibitors against multidrug resistant S. pneumoniae.

  6. Chlamydophila psittaci genotype E/B transmission from African grey parrots to humans.

    Science.gov (United States)

    Harkinezhad, Taher; Verminnen, Kristel; Van Droogenbroeck, Caroline; Vanrompay, Daisy

    2007-08-01

    Thirty-six birds from a parrot relief and breeding centre, as well as the manager, were examined for the presence of Chlamydophila psittaci. In the relief unit, 5 of 20 African grey parrots showed depression, ruffled feathers, loss of weight and mild dyspnoea. The birds received no antibiotic treatment. Birds of the breeding unit, 14 blue and gold macaws and 2 green-winged macaws, were healthy. They received doxycycline at the start of each breeding season. The manager complained of shortness of breath but took no medication. Using a nested PCR enzyme immunoassay (EIA), Cp. psittaci was detected in the faeces of all five sick birds, as well as in a nasal and pharyngeal swab from the manager. The veterinarian and her assistant became infected while sampling the parrots, as pharyngeal and nasal swabs from both were positive by nested PCR/EIA after visiting the parrot relief and breeding centre, but they showed no clinical signs of infection. Bacteria could be isolated from three of five nested PCR/EIA-positive birds, the manager and the veterinarian, but not from the veterinary assistant. Using an ompA genotype-specific real-time PCR, Cp. psittaci genotype E/B was identified as the transmitted strain. All breeding birds tested negative for Cp. psittaci. This is believed to be the first report on Cp. psittaci genotype E/B transmission from parrots to humans. In contradiction to genotype A strains, which are thought to be highly virulent to both birds and men, the currently described genotype E/B strain apparently caused no severe clinical symptoms in either parrots or humans.

  7. 鹦鹉热嗜衣原体PCR检测体系的建立%Development of Polymerase Chain Reaction to Detect Chlamydophila psittaci

    Institute of Scientific and Technical Information of China (English)

    柳陈坚; 龚福明; 蔡燕; 李海燕; 张忠华

    2009-01-01

    鹦鹉热是由鹦鹉热嗜衣原体(Chlamydophila psittaci,C. psittaci)引起,并可由带病鸟类传染给人类的一种人畜共患传染病.虽然有巢式PCR和多重PCR被开发用于C.psittaci的检测,但是这些方法较为繁琐.本研究应用了32株衣原体和嗜衣原体菌株,24株来自鸟类及人的肠道与呼吸道的细菌菌株,此外还使用606个禽鸟粪便、泄殖腔棉拭子及人体气管分泌物的临床样本.笔者以C.psittaci的MOMP基因(major outer membrane protein gene)为靶基因而开发了PCR检测方法.该最低能检测到100fgDNA(相当于10个衣原体)相当的鹦鹉热嗜衣原体.该靶基因区域涵盖了除C.pneumoniae、C.pecorum、C. trachomatis、C.suis和C.muridarum以外的鹦鹉热嗜衣原体所有的菌株及其他部分嗜衣原体菌株.606例临床样本中有29例鸟类粪便样本、25例泄殖腔棉拭子样本和2例人源样长检测为阳性.结果表明,该PCR检测法是检测鸟源鹦鹉热嗜衣原体的快速、敏感、有效的方法.

  8. Preoperative concentrations of suPAR and MBL proteins are associated with the development of pneumonia after elective surgery for colorectal cancer

    DEFF Research Database (Denmark)

    Svendsen, Mads N.; Ytting, Henriette; Brünner, Nils

    2006-01-01

    of postoperative infectious complications. The aim of this retrospective study was to determine the association between the soluble uPA receptor (suPAR) and MBL concentrations and the development of postoperative bacterial infectious complications. METHODS: Blood samples were drawn preoperatively from 544 patients...... or both, intra-abdominal abscess, anastomotic leakage, pneumonia, and blood stream infection. Data on perioperative blood transfusions in addition to clinical baseline characteristics were included as well. RESULTS: The numbers of surgical site infections, intra-abdominal abscesses, anastomotic leakages...... transfusion (p = 0.006; OR 1.5; 95% CI 1.1, 2.0) were associated with the development of pneumonia. Significant associations with other bacterial infections could not be demonstrated. Multivariate analysis including disease stage, sex, and age showed that suPAR, MBL, and perioperative blood transfusion were...

  9. Streptococcus pneumoniae fructose-1,6-bisphosphate aldolase, a protein vaccine candidate, elicits Th1/Th2/Th17-type cytokine responses in mice.

    Science.gov (United States)

    Elhaik Goldman, Shirin; Dotan, Shahar; Talias, Amir; Lilo, Amit; Azriel, Shalhevet; Malka, Itay; Portnoi, Maxim; Ohayon, Ariel; Kafka, Daniel; Ellis, Ronald; Elkabets, Moshe; Porgador, Angel; Levin, Ditza; Azhari, Rosa; Swiatlo, Edwin; Ling, Eduard; Feldman, Galia; Tal, Michael; Dagan, Ron; Mizrachi Nebenzahl, Yaffa

    2016-04-01

    Streptococcus pneumoniae (S. pneumoniae) is a major pathogen worldwide. The currently available polysaccharide-based vaccines significantly reduce morbidity and mortality. However, the inherent disadvantages of the currently available polysaccharide-based vaccines have motivated the search for other bacterial immunogens capable of eliciting a protective immune response against S. pneumoniae. Fructose-1,6-bisphosphate aldolase (FBA) is a glycolytic enzyme, which was found to localize to the bacterial surface, where it functions as an adhesin. Previously, immunizing mice with recombinant FBA (rFBA) in the presence of alum elicited a protective immune response against a lethal challenge with S. pneumoniae. Thus, the aim of the present study was to determine the cytokine responses that are indicative of protective immunity following immunization with rFBA. The protective effects against pneumococcal challenge in mice immunized with rFBA with complete Freund's adjuvant (CFA) in the initial immunization and with incomplete Freund's adjuvant (IFA) in booster immunizations surpassed the protective effects observed following immunization with either rFBA + alum or pVACfba. CD4+ T-cells obtained from the rFBA/CFA/IFA/IFA-immunized mice co-cultured with rFBA-pulsed antigen-presenting cells (APCs), exhibited a significantly greater proliferative ability than CD4+ T-cells obtained from the adjuvant-immunized mice co-cultured with rFBA‑pulsed APCs. The levels of the Th1-type cytokines, interferon (IFN)-γ, interleukin (IL)-2, tumor necrosis factor (TNF)-α and IL-12, the Th2-type cytokines, IL-4, IL-5 and IL-10, and the Th17-type cytokine, IL-17A, significantly increased within 72 h of the initiation of co-culture with CD4+ T-cells obtained from the rFBA‑immunized mice, in comparison with the co-cultures with CD4+ T-cells obtained from the adjuvant-immunized mice. Immunizing mice with rFBA resulted in an IgG1/IgG2 ratio of 41, indicating a Th2 response with substantial Th1

  10. Monoclonal Idiotope Vaccine against Streptococcus pneumoniae Infection

    Science.gov (United States)

    McNamara, Mary K.; Ward, Ronald E.; Kohler, Heinz

    1984-12-01

    A monoclonal anti-idiotope antibody coupled to a carrier protein was used to immunize BALB/c mice against a lethal Streptococcus pneumoniae infection. Vaccinated mice developed a high titer of antibody to phosphorylcholine, which is known to protect against infection with Streptococcus pneumoniae. Measurement of the median lethal dose of the bacteria indicated that anti-idiotope immunization significantly increased the resistance of BALB/c mice to the bacterial challenge. Antibody to an idiotope can thus be used as an antigen substitute for the induction of protective immunity.

  11. Prevalência de anticorpos anti-Chlamydophila spp. em propriedades rurais com histórico de aborto bovino no estado do Paraná Prevalence of antibodies against Chlamydophila spp. in herds with bovine abortion of Paraná state, Brazil

    Directory of Open Access Journals (Sweden)

    Francielle G. Silva-Zacarias

    2009-03-01

    Full Text Available Chlamydophila abortus é o agente etiológico do aborto epizoótico bovino, cujas manifestações clínicas mais freqüentes são aborto, nascimento de bezerros prematuros e de animais fracos, natimorto e repetição de cio em intervalos irregulares. O objetivo deste trabalho foi estimar a prevalência de anticorpos anti-Chlamydophila spp. em fêmeas bovinas de propriedades rurais com histórico de aborto, selecionadas dentro do delineamento amostral do Plano Nacional de Controle e Erradicação da Brucelose e Tuberculose no estado do Paraná. Foram testadas pela prova de fixação de complemento 3.102 amostras de soro de fêmeas bovinas (idade > 24 meses, provenientes de 373 propriedades. Ao total, 44 (1,42% animais foram positivos com títulos > 32. A prevalência de focos foi de 8,82% (6,15%-12,17%. Animais confinados ou semi-confinados (OR=3.339, P=0.004, propriedade com menos de 35 matrizes (OR=3.339, P=0.017, presença de produtos do aborto na pastagem (OR=2.372, P=0.037 e aluguel de pasto (OR=3.398, P=0.006 foram considerados fatores de risco para Chlamydophila spp. A infecção por Chlamydophila spp. acometeu um número pequeno de animais, oriundos de propriedades com histórico de aborto. A importância deste agente como causa de aborto em bovinos no estado do Paraná, se existir, é muito pequena.Chlamydophila abortus is a recognized cause of bovine epizootic abortion. Abortion, premature birth and weak lamb/calf, stillbirth and repeat breeding in irregular intervals are the most frequent disease manifestations. The complement fixation test is the recommended by the World Organization of Animal Health (OIE for Chlamydophila spp. serologic diagnosis. The aim of this study was estimate the prevalence of antibodies against Chlamydophila spp. in cattle herds with abortion, selected inside the sampling design of National Program of Control and Erradication of Brucellosis in Paraná state. Serum samples of 3,102 cows (age > 24 months from

  12. Hospital-acquired pneumonia

    Science.gov (United States)

    ... tends to be more serious than other lung infections because: People in the hospital are often very sick and cannot fight off ... prevent pneumonia. Most hospitals have programs to prevent hospital-acquired infections.

  13. Pneumonia (For Parents)

    Science.gov (United States)

    ... Development Infections Diseases & Conditions Pregnancy & Baby Nutrition & Fitness Emotions & Behavior School & Family Life First Aid & Safety Doctors & ... to dehydration in extreme cases, bluish or gray color of the lips and fingernails If the pneumonia ...

  14. Purification of a Mycoplasma pneumoniae adhesin by monoclonal antibody affinity chromatography.

    OpenAIRE

    Leith, D K; Baseman, J B

    1984-01-01

    A 165,000-dalton surface protein of Mycoplasma pneumoniae, designated protein P1, appears to be the major attachment ligand of the pathogen. We employed monoclonal antibody affinity chromatography to obtain purified protein P1.

  15. 肺炎嗜衣原体MOMP和人IL-2融合基因DNA疫苗的免疫原性研究%Immunogenicity of the DNA vaccine with monogene and fusion gene of the major outer membrane protein in Chlamydia pneumoniae and human IL-2

    Institute of Scientific and Technical Information of China (English)

    谢长青; 吴移谋; 曾焱华; 陈虹亮; 游晓星; 周洲

    2009-01-01

    目的 构建肺炎嗜衣原体(Chlamydophila pneumoniae,Cpn)主要外膜蛋白(MOMP)单基因及momp和人IL-2融合基因的真核表达载体并比较其免疫反应性,为研制Cpn核酸疫苗提供理论和实验依据. 方法 扩增Cpn momp及人IL-2基因并将二者融合,将momp和momp-IL-2分别克隆至pcDNA3.1(+)真核表达载体;制备纳米粒DNA疫苗免疫BALB/c小鼠,免疫荧光组化法检测重组质粒在小鼠组织内的稳定表达;ELISA法检测小鼠血清中的特异性抗体和小鼠脾细胞培养上清中IFN-γ水平;MTT法测定脾淋巴细胞特异性增殖反应. 结果 成功制备了pcDNA3.1(+)-momp和pcDNA3.1(+)-momp-IL-2的纳米核酸疫苗;Cpn momp单基因和momp-IL-2融合基因核酸疫苗均能刺激小鼠产生特异性抗体,且pcDNA3.1(+)-momp-IL-2能诱导小鼠产生更高效价的特异性抗体(P<0.05);pcDNA3.1(+)-momp-IL-2诱导小鼠脾细胞产生IFN-γ的量及对特异抗原的刺激指数明显高于pcDNA3.1(+)-momp.结论 Cpn momp单基因核酸疫苗和momp-IL-2融合基因核酸疫苗均能刺激小鼠产生较强的体液免疫和细胞免疫应答;pcDNA3.1(+)-momp-IL-2的免疫效果强于pcDNA3.1(+)-momp.

  16. Influence of Klebsiella pneumoniae CRP protein on bacterial adhesion and virulence in vitro%肺炎克雷伯菌转录调控子CRP对菌株粘附能力及细胞活性的影响

    Institute of Scientific and Technical Information of China (English)

    谭斌; 白群华; 罗美; 杨世亚; 薛健; 周锡鹏; 李迎丽; 邱景富

    2014-01-01

    Objective To analyze the adhesion and cell virulence of Klebsiella pneumonia wild type (WT) strain,complemented strain c-Δcrp (cAMP receptor protein) and mutant strain Δcrp,in order to investigate crp gene on the adhesion and cell toxicity of Klebsiella pneumonia.Methods After infection of A549 cells by Klebsiella pneumonia WT strain,c-Δcrp strain and Δcrp strain,the cells were lysed and the bacteria were quantified by plating appropriate dilutions on Luria-Bertani agar plates.LDH release was detected to estimate cell activity.Infection time and MOI were optimized.Results The adhesion ability of Klebsiella pneumonia WT (logCFU =5.145) and c-Δcrp strain (logCFU =4.915) was higher than that of Δcrp strain (logCFU =4.122) (P =0.004).The optimal conditions to determinate the LDH release included infected cells incubation for 8 h at 37 ℃,the developing time for 10 min in dark,and 1:10 dilution of the supernatant for test.The virulence of WT strain (70.69%) was significantly higher than that of Δcrp strain (19.54%) (P=0.001).Conclusion Knocking-out of crp gene causes obvious decrease of cellular toxicity and adhesion,comparing with the WT strain and c-Δcrp strain.Klebsiella pneumonia CRP protein positively regulates bacterial virulence and adhesion.%目的 分析肺炎克雷伯菌临床分离株WT(wild type)、回补株(c-Δcrp)和突变株(Δcrp)对人肺癌上皮细胞A549细胞的粘附能力及细胞活性的影响.方法 肺炎克雷伯菌WT株、c-Δcrp株和Δcrp株感染人肺癌上皮细胞A549,经裂解液裂解后平板计数计算粘附的菌量.LDH释放法检测细菌对细胞的毒性,优化感染时间和感染指数.结果 WT株及c-Δcrp株粘附的菌量分别为logCFU=5.145和logCFU=4.915,均高于Δcrp株(logCFU=4.122),差异有统计学意义(F=8.366,P=0.004).以MOI=1 000(细菌∶细胞=1000)的菌量感染靶细胞,37℃孵育8h,加底物液避光显色10 min,离心所得上清稀释10倍进行测定为最佳反应条件.WT

  17. Rapid detection of Chlamydia/Chlamydophila group in samples collected from swine herds with and without reproductive disorders.

    Science.gov (United States)

    Rypula, K; Kumala, A; Lis, P; Niemczuk, K; Płoneczka-Janeczko, K; Pejsak, Z

    2014-01-01

    The study was carried out in seven reproductive herds of pigs. In three of them reproductive disorders were observed. Three herds consisted of 10-50 and four consisted of 120-500 adult sows and they were called small and medium, respectively. Fifty-seven adult sows were randomly selected from herds. Serum samples were tested using the complement fixation test and swabs from both eyes and from the vaginal vestibule were examined using real-time PCR. All serum samples were negative. Infected sows were present in each of the study herds. In total, there were 28 positive samples (53%, 28/48) in real-time PCR in sows with reproductive disorders and 35 (53%, 35/66) in sows selected from herds without problems in reproduction. One isolate proved to be Chlamydophila pecorum, whereas all the remaining were Chamydia suis.

  18. Detection of all Chlamydophila and Chlamydia spp. of veterinary interest using species-specific real-time PCR assays.

    Science.gov (United States)

    Pantchev, Alexandra; Sting, Reinhard; Bauerfeind, Rolf; Tyczka, Judith; Sachse, Konrad

    2010-12-01

    The aim of the present study was to analyse the occurrence of chlamydiae in several mammalian host species. Clinical samples that previously tested positive in a Chlamydiaceae-specific real-time PCR were retested using six species-specific real-time PCR assays to identify the chlamydial species involved. Chlamydophila (Cp.) abortus was the agent most frequently found in cattle, sheep, horses, goats, and pigs. Detection in cattle of Cp. psittaci (11% of samples) and Chlamydia (C.) suis (9%), as well as Cp. psittaci in a goat sample was somewhat unexpected. DNA of two different chlamydiae was identified in 56 (12.7%) of 440 samples tested. Cp. felis was the predominant species found in cats, while in guinea pigs and rabbits only Cp. caviae was detected. Interestingly, the latter two pathogens were also identified in samples from dogs. The data show that mixed chlamydial infections are not rare and suggest an extended host range of individual species.

  19. Real-time detection and identification of Chlamydophila species in veterinary specimens by using SYBR green-based PCR assays.

    Science.gov (United States)

    Nordentoft, Steen; Kabell, Susanne; Pedersen, Karl

    2011-09-01

    Infections caused by members of the Chlamydiaceae family have long been underestimated due to the requirement of special laboratory facilities for the detection of this group of intracellular pathogens. Furthermore, new studies of this group of intracellular pathogens have revealed that host specificity of different species is not as clear as recently believed. As most members of the genus Chlamydophila have shown to be transmissible from animals to humans, sensitive and fast detection methods are required. In this study, SYBR green-based real-time assays were developed that detect all members of Chlamydiaceae and differentiate the most prevalent veterinary Chlamydophila species: Cp. psittaci, Cp. abortus, Cp. felis, and Cp. caviae. By adding bovine serum albumin to the master mixes, target DNA could be detected directly in crude lysates of enzymatically digested conjunctival or pharyngeal swabs or tissue specimens from heart, liver, and spleen without further purification. The assays were evaluated on veterinary specimens where all samples were screened using a family-specific PCR, and positive samples were further tested using species-specific PCRs. Cp. psittaci was detected in 47 birds, Cp. felis was found in 10 cats, Cp. caviae was found in one guinea pig, and Cp. abortus was detected in one sheep. The screening assay appeared more sensitive than traditional microscopical examination of stained tissue smears. By combining a fast, robust, and cost-effective method for sample preparation with a highly sensitive family-specific PCR, we were able to screen for Chlamydiaceae in veterinary specimens and confirm the species in positive samples with additional PCR assays.

  20. How Can Pneumonia Be Prevented?

    Science.gov (United States)

    ... t last as long Fewer serious complications Pneumococcal pneumonia vaccines Two vaccines are available to prevent pneumococcal ... Vaccination Web page. Other ways to help prevent pneumonia You also can take the following steps to ...

  1. Factors associated with upper respiratory tract disease caused by feline herpesvirus, feline calicivirus, Chlamydophila felis and Bordetella bronchiseptica in cats: experience from 218 European catteries

    OpenAIRE

    Helps, C. R.; Lait, P.; Damhuis, A.; Björnehammar, U.; Bolta, D.; Brovida, C.; Chabanne, L.; Egberink, H; Ferrand, G.; Fontbonne, A.; Pennisi, M G; Gruffydd-Jones, T.; Gunn-Moore, D.; Hartmann, K.; Lutz, H

    2005-01-01

    A full history of the management practices and the prevalence of upper respiratory tract disease (URTD) at 218 rescue shelters, breeding establishments and private households with five or more cats was recorded. Oropharyngeal and conjunctival swabs and blood samples were taken from 1748 cats. The prevalences of feline herpesvirus (FHV), feline calicivirus (FCV), Chlamydophila felis and Bordetella bronchiseptica were determined by PCR on swab samples. An ELISA was applied to determine the prev...

  2. 降钙素原与 C-反应蛋白联合检测在成人细菌性肺炎中的临床评价%Clinical evaluation of combined detection of serum procalcitonin and C-reactive protein for bacterial pneumonia in adults

    Institute of Scientific and Technical Information of China (English)

    曹校校; 李强; 陈荣

    2014-01-01

    OBJECTIVE To evaluate clinical effect of combined detection of serum procalcitonin (PCT ) and C-reactive protein (CRP) on diagnosis and treatment of bacterial pneumonia in adults in order to facilitate diagnosis and treatment for diseases . METHODS Patients with bacterial pneumonia , viral pneumonia and mycoplasma pneumonia of each 60 cases treated from Jan .2013 to Jun .2013 in our hospital were randomly selected and respectively received combined PCT and CRP detection .The 60 patients with bacterial pneumonia were grouped according to the severity of illness .PCT and CRP contents in patients of each group were summarized and the data were processed by statistical software SPSS17 .0 .RESULTS PTC of pneumonic patients was (9 .31 ± 4 .76)μg/L in the bacterial pneumonia group ,(1 .30 ± 0 .68)μg/L in the viral pneumonia group and (1 .45 ± 0 .87)μg/L in the mycoplasma pneumonia group .CRP was (64 .21 ± 16 .97) mmol/L in the bacterial pneumonia group ,(30 .05 ± 10 .02) mmol/L in the mycoplasma pneumonia group and (4 .79 ± 1 .22) mmol/L in the viral pneumonia group . The difference between the three groups was significant (P<0 .05) .The level of PCT and CRP increased along with the severity of bacterial pneumonia .CONCLUSION PCT and CRP levels are closely related with bacterial pneumonia ,the combined detection of the two indexes can provide good guidance for diagnosis and treatment as well as prognosis for bacterial pneumonia in adults .%目的:探讨血清降钙素原(PCT)和C-反应蛋白(CRP)联合检测在成人细菌性肺炎诊断与治疗中的临床评价,以利于疾病的诊断治疗。方法随机选取2013年1-6月进行治疗的细菌性肺炎、病毒性肺炎、支原体肺炎患者各60例,分别对其进行PCT和CRP联合的检测,将60例细菌性肺炎患者按病情轻重程度分组,统计各组PCT和CRP含量,采用SPSS17.0统计软件对数据进行处理。结果肺炎患者PCT水平细菌性肺炎组为(9.31±4

  3. Secretion of Cpn0796 from Chlamydia pneumoniae into the host cell cytoplasm by an autotransporter mechanism

    DEFF Research Database (Denmark)

    Vandahl, Brian B S; Stensballe, Allan; Roepstorff, Peter

    2005-01-01

    By comparison of proteome profiles of purified Chlamydia pneumoniae and whole lysates of C. pneumoniae infected HEp-2 cells, an N-terminal fragment of the previously uncharacterized chlamydial protein Cpn0796 was identified as a secreted protein. A 38 kDa cleavage product of Cpn0796 was present i...

  4. IL-17A is proatherogenic in high-fat diet-induced and Chlamydia pneumoniae infection-accelerated atherosclerosis in mice.

    Science.gov (United States)

    Chen, Shuang; Shimada, Kenichi; Zhang, Wenxuan; Huang, Ganghua; Crother, Timothy R; Arditi, Moshe

    2010-11-01

    The role of IL-17 in atherogenesis remains controversial. We previously reported that the TLR/MyD88 signaling pathway plays an important role in high-fat diet as well as Chlamydophila pneumoniae infection-mediated acceleration of atherosclerosis in apolipoprotein E-deficient mice. In this study, we investigated the role of the IL-17A in high-fat diet (HFD)- and C. pneumoniae-induced acceleration of atherosclerosis. The aortic sinus plaque and aortic lesion size and lipid composition as well as macrophage accumulation in the lesions were significantly diminished in IL-17A(-/-) mice fed an HFD compared with wild-type (WT) C57BL/6 control mice. As expected, C. pneumoniae infection led to a significant increase in size and lipid content of the atherosclerotic lesions in WT mice. However, IL-17A(-/-) mice developed significantly less acceleration of lesion size following C. pneumoniae infection compared with WT control despite similar levels of blood cholesterol levels. Furthermore, C. pneumoniae infection in WT but not in IL-17A(-/-) mice was associated with significant increases in serum concentrations of IL-12p40, CCL2, IFN-γ, and numbers of macrophages in their plaques. Additionally, in vitro studies suggest that IL-17A activates vascular endothelial cells, which secrete cytokines that in turn enhance foam cell formation in macrophages. Taken together, our data suggest that IL-17A is proatherogenic and that it plays an important role in both diet-induced atherosclerotic lesion development, and C. pneumoniae infection-mediated acceleration of atherosclerotic lesions in the presence of HFD.

  5. Lung Dendritic Cells Facilitate Extrapulmonary Bacterial Dissemination during Pneumococcal Pneumonia

    Directory of Open Access Journals (Sweden)

    Alva eRosendahl

    2013-06-01

    Full Text Available Streptococcus pneumoniae is a leading cause of bacterial pneumonia worldwide. Given the critical role of dendritic cells (DCs in regulating and modulating the immune response to pathogens, we investigated here the role of DCs in S. pneumoniae lung infections. Using a well-established transgenic mouse line which allows the conditional transient depletion of DCs, we showed that ablation of DCs resulted in enhanced resistance to intranasal challenge with S. pneumoniae. DC-depleted mice exhibited delayed bacterial systemic dissemination, significantly reduced bacterial loads in the infected organs and lower levels of serum inflammatory mediators than non-depleted animals. The increased resistance of DC-depleted mice to S. pneumoniae was associated with a better capacity to restrict pneumococci extrapulmonary dissemination. Furthermore, we demonstrated that S. pneumoniae disseminated from the lungs into the regional lymph nodes in a cell-independent manner and that this direct way of dissemination was much more efficient in the presence of DCs. We also provide evidence that S. pneumoniae induces expression and activation of matrix metalloproteinase-9 (MMP-9 in cultured bone marrow-derived DCs. MMP-9 is a protease involved in the breakdown of extracellular matrix proteins and is critical for DC trafficking across extracellular matrix and basement membranes during the migration from the periphery to the lymph nodes. MMP-9 was also significantly up-regulated in the lungs of mice after intranasal infection with S. pneumoniae. Notably, the expression levels of MMP-9 in the infected lungs were significantly decreased after depletion of DCs suggesting the involvement of DCs in MMP-9 production during pneumococcal pneumonia. Thus, we propose that S. pneumoniae can exploit the DC-derived proteolysis to open tissue barriers thereby facilitating its own dissemination from the local site of infection.

  6. Bactericidal effect of bovine lactoferrin and synthetic peptide lactoferrin chimera in Streptococcus pneumoniae and the decrease in luxS gene expression by lactoferrin

    NARCIS (Netherlands)

    N. León-Sicairos; U.A. Angulo-Zamudio; J.E. Vidal; C.A. López-Torres; J.G.M. Bolscher; K. Nazmi; R. Reyes-Cortes; M. Reyes-López; M. de la Garza; A. Canizalez-Román

    2014-01-01

    Streptococcus pneumoniae (pneumococcus) is responsible for nearly one million child deaths annually. Pneumococcus causes infections such as pneumonia, otitis media, meningitis, and sepsis. The human immune system includes antibacterial peptides and proteins such as lactoferrin (LF), but its activity

  7. Hypervirulent Klebsiella pneumoniae

    OpenAIRE

    Patel, Payal K.; Russo, Thomas A.; Karchmer, Adolf W.

    2014-01-01

    Hypervirulent strains of Klebsiella pneumoniae are associated with abscess formation, commonly hepatic, and metastatic spread, even in healthy patients. We describe a case of this clinical syndrome, genotypic and phenotypic features of the isolate, and briefly review epidemiology, clinical manifestations, and pathogenesis of this underappreciated syndrome.

  8. Bacteremia with Streptococcus pneumoniae

    DEFF Research Database (Denmark)

    Christensen, J S; Jensen, T G; Kolmos, H J

    2012-01-01

    We conducted a hospital-based cohort study among adult patients with first-time Streptococcus pneumoniae bacteremia (SPB) from 2000 through 2008. Patients were identified in a population-based bacteremia database and followed up for mortality through the Danish Civil Registration System (CRS...

  9. Perfil clínico, epidemiológico e etiológico de pacientes com pneumonia adquirida na comunidade internados em um hospital geral da microrregião de Sumaré, SP Clinical, epidemiological, and etiological profile of inpatients with community-acquired pneumonia at a general hospital in the Sumaré microregion of Brazil

    Directory of Open Access Journals (Sweden)

    Maria Rita Donalisio

    2011-04-01

    Full Text Available OBJETIVO: Analisar aspectos clínicos, etiológicos e epidemiológicos das pneumonias adquiridas na comunidade (PAC em indivíduos internados. MÉTODOS: Foram estudados prospectivamente 66 pacientes com PAC maiores de 14 anos no Hospital Estadual Sumaré, localizado na cidade de Sumaré (SP, entre outubro de 2005 e setembro de 2007. Coletamos dados sobre história clínica, exame clínico, escore pneumonia severity index (PSI e exames laboratoriais (hemocultura, bacterioscopia/cultura de escarro, sorologias para Chlamydophila pneumoniae, Mycoplasma pneumoniae e Legionella pneumophila, além de antígenos urinários de Legionella sp. e Streptococcus pneumoniae. RESULTADOS: A idade média dos pacientes foi de 53 anos, a maioria tinha baixa escolaridade, e 55,7% apresentavam pelo menos uma comorbidade no momento da internação. O percentual de idosos vacinados contra influenza entre os internados foi significativamente menor que os da comunidade dos municípios da microrregião de Sumaré (52,6% vs. > 70%. A febre foi menos frequente entre os idosos (p OBJECTIVE: To analyze the clinical, etiological, and epidemiological aspects of community-acquired pneumonia (CAP in hospitalized individuals. METHODS: We prospectively studied 66 patients (> 14 years of age with CAP admitted to the Hospital Estadual Sumaré, located in the Sumaré microregion of Brazil, between October of 2005 and September of 2007. We collected data related to clinical history, physical examination, pneumonia severity index (PSI scores, and laboratory tests (blood culture; sputum smear microscopy and culture; serology for Chlamydophila pneumoniae, Mycoplasma pneumoniae, and Legionella pneumophila; and detection of Legionella sp. and Streptococcus pneumoniae antigens in urine. RESULTS: The mean age of patients was 53 years. Most had a low level of education, and 55.7% presented with at least one comorbidity at the time of hospitalization. The proportion of elderly people vaccinated

  10. Nasopharyngeal carriage of Streptococcus pneumonia in pneumonia-prone age groups in Semarang, Java Island, Indonesia

    NARCIS (Netherlands)

    H. Farida (Helmia); J.A. Severin (Juliëtte); M.H. Gasem; M. Keuter (Monique); H. Wahyono (Hendro); P. van den Broek (Peterhans); P.W.M. Hermans (Peter); H.A. Verbrugh (Henri)

    2014-01-01

    textabstractIntroduction: Streptococcus pneumoniae is a worldwide occurring pathogen Nasopharyngeal carriage of Streptococcus pneumoniae precedes pneumonia and other pneumococcal diseases in the community. Little is known about S. pneumoniae carriage in Indonesia, complicating strategies to control

  11. Nasopharyngeal Carriage of Streptococcus pneumonia in Pneumonia-Prone Age Groups in Semarang, Java Island, Indonesia

    NARCIS (Netherlands)

    Farida, H.; Severin, J.A.; Gasem, M.H.; Keuter, M.; Wahyono, H.; Broek, P van den; Hermans, P.W.M.; Verbrugh, H.A.

    2014-01-01

    INTRODUCTION: Streptococcus pneumoniae is a worldwide occurring pathogen Nasopharyngeal carriage of Streptococcus pneumoniae precedes pneumonia and other pneumococcal diseases in the community. Little is known about S. pneumoniae carriage in Indonesia, complicating strategies to control pneumococcal

  12. Immune response to Mycoplasma pneumoniae P1 and P116 in patients with atypical pneumonia analyzed by ELISA

    Directory of Open Access Journals (Sweden)

    Birkelund Svend

    2004-02-01

    Full Text Available Abstract Background Serology is often used for the diagnosis of Mycoplasma pneumoniae. It is important to identify specific antigens that can distinguish between the presence or absence of antibodies against M. pneumoniae. The two proteins, P116 and P1, are found to be immunogenic. By using these in ELISA it is possible to identify an immune response against M. pneumoniae in serum samples. Results A recombinant protein derived from the P116 protein and one from the P1 protein were used in two ELISA tests, rP116-ELISA and rP1-ELISA. Human serum samples from patients with atypical pneumonia were tested and compared to the results of the complement fixation test. There was a good agreement between the two tests but the rP1-ELISA showed the best discrimination between positive and negative samples. Conclusion Two ELISA tests based on recombinant proteins have been analysed and compared to the complement fixation test results. The two ELISA tests were found suitable for use in serodiagnostics of M. pneumoniae infections. The use of specific antigens eliminates the risk of cross reaction to an immune response against other bacteria.

  13. 肺炎链球菌Tpx蛋白的原核表达、纯化及活性分析%Prokaryotic expression and purification of the Tpx protein from Streptococcus pneumoniae

    Institute of Scientific and Technical Information of China (English)

    王剑; 赵兰华; 李冉辉

    2017-01-01

    目的 原核表达、纯化肺炎链球菌(Streptococcus pneumoniae)Tpx蛋白并测定其活性. 方法 利用生物信息学方法分析Tpx蛋白的生物学特性.设计特异性引物,PCR扩增Tpx基因并对进行双酶切,酶切片段连接到表达质粒pET28a.将重组质粒转化大肠埃希菌BL21(DE3),利用IPTG诱导重组Tpx蛋白表达,使用Ni2+亲和层析进行纯化,采用氧化铁二甲酚橙法(FOX)测定其活性. 结果 生物信息学分析Tpx蛋白无信号肽,无跨膜结构域.α-螺旋、延伸主链、β-转角和无规卷曲的含量分别为30%、27.6%、11.6和30.7%.晶体结构在蛋白结构数据库(PDB)中的登陆号为1psq.1.A.PCR扩增Tpx基因片段大小为492 bp,连接到pET28a后获得重组质粒pET28a-Tpx.重组质粒转化DE3后经IPTG诱导表达分子质量单位约为22 ku的重组蛋白,经亲和层析得到高纯度的重组Tpx蛋白,该蛋白能分解H2O2和叔丁基过氧化氢(t-BHP). 结论 成功构建pET28a-Tpx质粒,表达的重组Tpx蛋白具有氧化酶活性,为研究肺炎链球菌的抗氧化机制奠定了实验基础.%Objectives To express the thiol peroxidase (Tpx) protein of Streptococcus pneumoniae in a prokaryotic system and to purify that protein and to measure its peroxidase activity.Methods The biological characteristics of the Tpx protein were analyzed bioinformatically.The Tpx gene was amplified with PCR using specific primers,which were double-digested,and then the gene was ligated into a pET-28.a vector.The recombinant plasmid was transformed into E.coli BL21 (DE3).Expression of the recombinant protein was induced with IPTG,and the protein was purified using Ni2 +-affinity chromatography.The peroxidase activity of Tpx was measured using a ferrous oxidation-xylenol orange assay.Results Bioinformatic results indicated that the Tpx protein has no signal peptides and that it lacks a transmembrane domain.Alpha helices accounted for 30% of its secondary structure,extended strands accounted

  14. Identification of two novel genes encoding 97- to 99-kilodalton outer membrane proteins of Chlamydia pneumoniae.Infect Immun. 1999 Jan;67(1):375-83

    DEFF Research Database (Denmark)

    Knudsen, K; Madsen, AS; Mygind, P

    1999-01-01

    of putative outer membrane proteins encoded by the Chlamydia psittaci and Chlamydia trachomatis gene families. By use of a monospecific polyclonal antibody against purified recombinant Omp4, it was shown that without heating, the protein migrated at 65 to 75 kDa in sodium dodecyl sulfate-polyacrylamide gel...

  15. Osteopontin promotes host defense during Klebsiella pneumoniae-induced pneumonia.

    Science.gov (United States)

    van der Windt, G J W; Hoogerwerf, J J; de Vos, A F; Florquin, S; van der Poll, T

    2010-12-01

    Klebsiella pneumoniae is a common cause of nosocomial pneumonia. Osteopontin (OPN) is a phosphorylated glycoprotein involved in inflammatory processes, some of which is mediated by CD44. The aim of this study was to determine the role of OPN during K. pneumoniae-induced pneumonia. Wild-type (WT) and OPN knockout (KO) mice were intranasally infected with 10⁴ colony forming units of K. pneumoniae, or administered Klebsiella lipopolysaccharides (LPS). In addition, recombinant OPN (rOPN) was intranasally administered to WT and CD44 KO mice. During Klebsiella pneumonia, WT mice displayed elevated pulmonary and plasma OPN levels. OPN KO and WT mice showed similar pulmonary bacterial loads 6 h after infection; thereafter, Klebsiella loads were higher in lungs of OPN KO mice and the mortality rate in this group was higher than in WT mice. Early neutrophil recruitment into the bronchoalveolar space was impaired in the absence of OPN after intrapulmonary delivery of either Klebsiella bacteria or Klebsiella LPS. Moreover, rOPN induced neutrophil migration into the bronchoalveolar space, independent from CD44. In vitro, OPN did not affect K. pneumoniae growth or neutrophil function. In conclusion, OPN levels were rapidly increased in the bronchoalveolar space during K. pneumoniae pneumonia, where OPN serves a chemotactic function towards neutrophils, thereby facilitating an effective innate immune response.

  16. Pneumocystis jirovecii pneumonia

    DEFF Research Database (Denmark)

    Cordonnier, Catherine; Cesaro, Simone; Maschmeyer, Georg

    2016-01-01

    Pneumocystis jirovecii can cause life-threatening pneumonia following treatment for haematological malignancies or after HSCT. The mortality rate of P. jirovecii pneumonia (PCP) in these patients is 30%-60%, especially after HSCT. The clinical presentation of PCP in haematology differs from...... and monoclonal antibodies in various haematological diseases, justifies constant vigilance in order to identify new at-risk populations and give prophylaxis accordingly. The fifth and sixth European Conferences on Infections in Leukaemia (ECIL-5 and ECIL-6) aimed to review risk factors for PCP in haematology...... patients and to establish evidence-based recommendations for PCP diagnosis, prophylaxis and treatment. This article focuses on the magnitude of the problem, the main differences in clinical presentation between haematology patients and other immunocompromised populations, especially HIV-infected patients...

  17. Electrocardiogram in pneumonia.

    Science.gov (United States)

    Stein, Paul D; Matta, Fadi; Ekkah, Maan; Saleh, Tarek; Janjua, Muhammad; Patel, Yash R; Khadra, Helmi

    2012-12-15

    Findings on electrocardiogram may hint that pulmonary embolism (PE) is present when interpreted in the proper context and lead to definitive imaging tests. However, it would be useful to know if electrocardiographic (ECG) abnormalities also occur in patients with pneumonia and whether these are similar to ECG changes with PE. The purpose of this investigation was to determine ECG findings in patients with pneumonia. We retrospectively evaluated 62 adults discharged with a diagnosis of pneumonia who had no previous cardiopulmonary disease and had electrocardiogram obtained during hospitalization. The most prevalent ECG abnormality, other than sinus tachycardia, was minor nonspecific ST-segment or T-wave changes occurring in 13 of 62 (21%). Right atrial enlargement occurred in 4 of 62 (6.5%). QRS abnormalities were observed in 24 of 62 (39%). Right-axis deviation and S(1)S(2)S(3) were the most prevalent QRS abnormalities, which occurred in 6 of 62 (9.7%). Complete right bundle branch block and S(1)Q(3)T(3) pattern occurred in 3 of 62 (4.8%). ECG abnormalities that were not present within 1 month previously or abnormalities that disappeared within 1 month included left-axis deviation, right-axis deviation, right atrial enlargement, right ventricular hypertrophy, S(1)S(2)S(3), S(1)Q(3)T(3), low-voltage QRS complexes, and nonspecific ST-segment or T-wave abnormalities. In conclusion, electrocardiogram in patients with pneumonia often shows QRS abnormalities or nonspecific ST-segment or T-wave changes. ECG findings are similar to ECG abnormalities in PE and electrocardiogram cannot assist in the differential diagnosis.

  18. Pediatric Round Pneumonia

    Directory of Open Access Journals (Sweden)

    Yen-Lin Liu

    2014-12-01

    Full Text Available “Round pneumonia” or “spherical pneumonia” is a well-characterized clinical entity that seems to be less addressed by pediatricians in Taiwan. We herein report the case of a 7-year-old boy who presented with prolonged fever, cough, and chest X-rays showing a well-demarcated round mass measuring 5.9 × 5.6 × 4.3 cm in the left lower lung field, findings which were typical for round pneumonia. The urinary pneumococcal antigen test was positive, and serum anti-Mycoplasma pneumoniae antibody titer measurement using a microparticle agglutination method was 1:160 (+. After oral administration of antibiotics including azithromycin and amoxicillin/clavulanate, which was subsequently replaced by ceftibuten due to moderate diarrhea, the fever subsided 2 days later and the round patch had completely resolved on the 18th day after the diagnosis. Recent evidence suggests treating classical round pneumonia with antibiotics first and waiving unwarranted advanced imaging studies, while alternative etiologies such as abscesses, tuberculosis, nonbacterial infections, congenital malformations, or neoplasms should still be considered in patients with atypical features or poor treatment response.

  19. Bronchiolitis Obliterans with Organizing Pneumonia (BOOP)

    Science.gov (United States)

    ... What can you tell me about cryptogenic organizing pneumonia? Answers from Teng Moua, M.D. Previously called bronchiolitis obliterans with organizing pneumonia, cryptogenic organizing pneumonia (COP) is a rare lung ...

  20. Pneumonia bacteriana adquirida na comunidade

    OpenAIRE

    Machado, Lais Del Prá Netto

    2015-01-01

    Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro de Ciências da Saúde, Programa de Pós-Graduação em Farmácia, Florianópolis, 2015. A pneumonia pode ser causada por diversos microrganismos e classificada de forma abrangente, havendo poucos e frágeis estudos clínicos e epidemiológicos sobre pneumonias adquiridas na comunidade (PACs). Os patógenos mais frequentes nas PACs são Streptococcus pneumoniae e Haemophilus influenzae (em pneumonias típicas) e Mycoplasma pneumoni...

  1. Burden of Severe Pneumonia, Pneumococcal Pneumonia and Pneumonia Deaths in Indian States: Modelling Based Estimates.

    Science.gov (United States)

    Farooqui, Habib; Jit, Mark; Heymann, David L; Zodpey, Sanjay

    2015-01-01

    The burden of severe pneumonia in terms of morbidity and mortality is unknown in India especially at sub-national level. In this context, we aimed to estimate the number of severe pneumonia episodes, pneumococcal pneumonia episodes and pneumonia deaths in children younger than 5 years in 2010. We adapted and parameterized a mathematical model based on the epidemiological concept of potential impact fraction developed CHERG for this analysis. The key parameters that determine the distribution of severe pneumonia episode across Indian states were state-specific under-5 population, state-specific prevalence of selected definite pneumonia risk factors and meta-estimates of relative risks for each of these risk factors. We applied the incidence estimates and attributable fraction of risk factors to population estimates for 2010 of each Indian state. We then estimated the number of pneumococcal pneumonia cases by applying the vaccine probe methodology to an existing trial. We estimated mortality due to severe pneumonia and pneumococcal pneumonia by combining incidence estimates with case fatality ratios from multi-centric hospital-based studies. Our results suggest that in 2010, 3.6 million (3.3-3.9 million) episodes of severe pneumonia and 0.35 million (0.31-0.40 million) all cause pneumonia deaths occurred in children younger than 5 years in India. The states that merit special mention include Uttar Pradesh where 18.1% children reside but contribute 24% of pneumonia cases and 26% pneumonia deaths, Bihar (11.3% children, 16% cases, 22% deaths) Madhya Pradesh (6.6% children, 9% cases, 12% deaths), and Rajasthan (6.6% children, 8% cases, 11% deaths). Further, we estimated that 0.56 million (0.49-0.64 million) severe episodes of pneumococcal pneumonia and 105 thousand (92-119 thousand) pneumococcal deaths occurred in India. The top contributors to India's pneumococcal pneumonia burden were Uttar Pradesh, Bihar, Madhya Pradesh and Rajasthan in that order. Our results

  2. Klebsiella pneumoniae secretes outer membrane vesicles that induce the innate immune response.

    Science.gov (United States)

    Lee, Je Chul; Lee, Eun Jeoung; Lee, Jung Hwa; Jun, So Hyun; Choi, Chi Won; Kim, Seung Il; Kang, Sang Sun; Hyun, Sunghee

    2012-06-01

    Outer membrane vesicles (OMVs) derived from pathogenic Gram-negative bacteria are an important vehicle for delivery of effector molecules to host cells, but the production of OMVs from Klebsiella pneumoniae, an opportunistic pathogen of both nosocomial and community-acquired infections, and their role in bacterial pathogenesis have not yet been determined. In the present study, we examined the production of OMVs from K. pneumoniae and determined the induction of the innate immune response against K. pneumoniae OMVs. Klebsiella pneumoniae ATCC 13883 produced and secreted OMVs during in vitro culture. Proteomic analysis revealed that 159 different proteins were associated with K. pneumoniae OMVs. Klebsiella pneumoniae OMVs did not inhibit cell growth or induce cell death. However, these vesicles induced expression of proinflammatory cytokine genes such as interleukin (IL)-1β and IL-8 in epithelial cells. An intratracheal challenge of K. pneumoniae OMVs in neutropenic mice resulted in severe lung pathology similar to K. pneumoniae infection. In conclusion, K. pneumoniae produces OMVs like other pathogenic Gram-negative bacteria and K. pneumoniae OMVs are a molecular complex that induces the innate immune response.

  3. Seroprevalence survey of Chlamydophila abortus infection in breeding goats on commercial farms in the Otavi Veterinary District, northern Namibia

    Directory of Open Access Journals (Sweden)

    Alaster Samkange

    2010-08-01

    Full Text Available A total of 1 076 sera from breeding goats were randomly collected from 24 different farms and tested with CHEKIT®-ELISA (IDEXX Laboratories B.V., 1 119 NE Schiphol-Rijk, Nederland for antibodies against Chlamydophila abortus. The farms were divided into two categories of twelve farms each,based on their previous history of observed abortions over the previous 12 months: those with low (< 5% levels of abortion and those with high (≥ 5% levels of abortion. The farmers were also interviewed on their level of awareness about chlamydophilosis, its zoonotic importance and vaccination measures against the disease. The study detected overall seroprevalence levels of 25% for the farms and 8% for the individual animals (at 95% confidence. A total of six out of twentyfour farms (25% had at least one positive breeding animal. Only five out of the twenty-four (20.8%farmers interviewed were aware of chlamydophilosis and its zoonotic dangers. None of the 24 farmers interviewed practised any vaccination against chlamydophilosis. There was a significantly higher number of seropositive animals from farms with high levels of abortion, compared to those animals from farms with low levels of abortion (p = 0.0001. This study underscores the need for a higher level of farmer awareness and training on chlamydophilosis and its zoonotic dangers.

  4. Seroprevalence and risk factors associated with Chlamydophila abortus infection in dairy goats in the Northeast of Brazil

    Directory of Open Access Journals (Sweden)

    Carolina S.A.B. Santos

    2012-11-01

    Full Text Available Few data are available on the prevalence and risk factors of Chlamydophila abortus infection in goats in Brazil. A cross-sectional study was carried out to determine the flock-level prevalence of C. abortus infection in goats from the semiarid region of the Paraíba State, Northeast region of Brazil, as well as to identify risk factors associated with the infection. Flocks were randomly selected and a pre-established number of female goats > 12 mo old were sampled in each of these flocks. A total of 975 serum samples from 110 flocks were collected, and structured questionnaire focusing on risk factors for C. abortus infection was given to each farmer at the time of blood collection. For the serological diagnosis the complement fixation test (CFT using C. abortus S26/3 strain as antigen was performed. The flock-level factors for C. abortus prevalence were tested using multivariate logistic regression model. Fifty-five flocks out of 110 presented at least one seropositive animal with an overall prevalence of 50.0% (95%; CI: 40.3%, 59.7%. Ninety-one out of 975 dairy goats examined were seropositive with titers >32, resulting in a frequency of 9.3%. Lend buck for breeding (odds ratio = 2.35; 95% CI: 1.04-5.33 and history of abortions (odds ratio = 3.06; 95% CI: 1.37-6.80 were associated with increased flock prevalence.

  5. Differential identification of Chlamydophila abortus live vaccine strain 1B and C. abortus field isolates by PCR-RFLP.

    Science.gov (United States)

    Laroucau, Karine; Vorimore, Fabien; Sachse, Konrad; Vretou, Evangelia; Siarkou, Victoria I; Willems, Hermann; Magnino, Simone; Rodolakis, Annie; Bavoil, Patrik M

    2010-08-09

    Comparative genomic analysis of a wild-type strain of the ovine pathogen Chlamydophila abortus and its nitrosoguanidine-induced, temperature-sensitive and virulence-attenuated live vaccine derivative identified point mutations unique to the mutant (Burall et al. [1]). Here, we evaluate the capacity of some of these mutations to either create or eliminate restriction sites using the wild-type strain C. abortus S26/3 as a reference. Three of eight genomic sites with confirmed point mutations (CAB153, CAB636 and CAB648) were retained for analysis as each resulted in the loss of a restriction site in the genome sequence of the vaccine strain. PCR-restriction fragment length polymorphism analysis using restriction enzymes chosen to specifically target the three genomic sites was then applied to a large number of C. abortus field isolates and reference strains. Our results indicate that the three mutations are uniquely present in the vaccine strain, and as such provide easy-to-use markers for the differential identification of the vaccine strain and wild-type isolates.

  6. Seroprevalence survey of Chlamydophila abortus infection in breeding goats on commercial farms in the Otavi Veterinary District, northern Namibia.

    Science.gov (United States)

    Samkange, Alaster; Katsande, Tendai C; Tjipura-Zaire, Georgina; Crafford, Jan E

    2010-08-20

    A total of 1 076 sera from breeding goats were randomly collected from 24 different farms and tested with CHEKIT®-ELISA (IDEXX Laboratories B.V., 1 119 NE Schiphol-Rijk, Nederland) for antibodies against Chlamydophila abortus. The farms were divided into two categories of twelve farms each,based on their previous history of observed abortions over the previous 12 months: those with low (< 5%) levels of abortion and those with high (≥ 5%) levels of abortion. The farmers were also interviewed on their level of awareness about chlamydophilosis, its zoonotic importance and vaccination measures against the disease. The study detected overall seroprevalence levels of 25% for the farms and 8% for the individual animals (at 95% confidence). A total of six out of twentyfour farms (25%) had at least one positive breeding animal. Only five out of the twenty-four (20.8%)farmers interviewed were aware of chlamydophilosis and its zoonotic dangers. None of the 24 farmers interviewed practised any vaccination against chlamydophilosis. There was a significantly higher number of seropositive animals from farms with high levels of abortion, compared to those animals from farms with low levels of abortion (p = 0.0001). This study underscores the need for a higher level of farmer awareness and training on chlamydophilosis and its zoonotic dangers.

  7. Detection of Chlamydophila caviae and Streptococcus equi subsp. zooepidemicus in horses with signs of rhinitis and conjunctivitis.

    Science.gov (United States)

    Gaede, Wolfgang; Reckling, Karl-Friedrich; Schliephake, Anette; Missal, Dirk; Hotzel, Helmut; Sachse, Konrad

    2010-05-19

    At a stud farm of Trakehner horses, all 33 foals of a birth cohort developed conjunctivitis and serous to muco-purulent rhinitis, and 7 older horses showed recurrent signs of conjunctivitis. Examination of nasal and conjunctival swabs by bacterial and cell culture, as well as real-time PCR, ArrayTube microarray analysis and DNA sequencing led to the identification of Chlamydophila (C.) caviae (first description in horses) and Streptococcus (S.) equi subsp. zooepidemicus. We presume a synergistic effect associated with these two agents by hypothesising that primary lesions were set by C. caviae and subsequently aggravated by Streptococcus equi subsp. zooepidemicus. Indications supporting this assumption include (i) the conjunctivitis caused by mono-infection with C. caviae, (ii) recurrent clinical symptoms in the affected animals, and (iii) the absence of a sustained clinical effect of antibiotic therapy with trimethoprim-sulfonamide, enrofloxacin and amoxicillin. The detection of C. caviae in horses raises questions about the significance and natural host range of this agent.

  8. 血清降钙素原和C反应蛋白水平改变在支原体肺炎患儿中的临床意义%Changes of serum procalcitonin and C - reactive protein levels in children with mycoplasma pneumonia

    Institute of Scientific and Technical Information of China (English)

    张月明

    2014-01-01

    目的:探讨血清降钙素原和 C 反应蛋白水平改变在支原体肺炎患儿中的临床意义。方法选择支原体肺炎、细菌性肺炎患儿各67例为支原体肺炎组和细菌性肺炎组,选择健康体检儿童67例为健康对照组。检测各组血清降钙素原(PCT)和 C 反应蛋白(CRP)水平并分析。结果支原体肺炎组急性期血清 PCT 水平显著高于健康对照组,但显著低于细菌性肺炎组( P 均<0.05)。支原体肺炎组、细菌性肺炎组血清 CRP 水平显著高于健康对照组( P <0.05),但两者 CRP 水平比较无显著差异。治疗后发现恢复期支原体肺炎组血清 PCT、CRP 水平显著下降,与健康对照组无统计学差异( P 均﹥0.05)。结论早期联合检测呼吸道感染患儿 PCT 与 CRP 水平可帮助早期鉴别支原体感染肺炎和细菌性感染肺炎,并可作为疗效观察的参考依据。%Objective To investigate the changes of serum procalcitonin(PCT)and C - reactive protein(CRP)levels in children with mycoplasma pneumonia. Methods Serum PCT and CRP levels in each group of children with mycoplasma pneumonia(n = 67)or bacterial pneu-monia(n = 67),and healthy children(n = 67)were detected and analyzed. Results In acute phase,serum PCT level was significantly higher in mycoplasma pneumonia group than healthy control group,but significantly lower than bacterial pneumonia group(all P < 0. 05). Serum CRP level in mycoplasma pneumonia group and bacterial pneumonia group was significantly higher than healthy control group( P < 0. 05),but no sig-nificant difference between mycoplasma pneumonia group and bacterial pneumonia group. After treatment,serum PCT and CRP levels were signifi-cantly decreased in children with mycoplasma pneumonia,but not significantly different from those of control group(all P ﹥ 0. 05). Conclusion Combined early detection of serum PCT and CRP for children with respiratory infection may help early identification

  9. Klebsiella pneumoniae FimK Promotes Virulence in Murine Pneumonia.

    Science.gov (United States)

    Rosen, David A; Hilliard, Julia K; Tiemann, Kristin M; Todd, Elizabeth M; Morley, S Celeste; Hunstad, David A

    2016-02-15

    Klebsiella pneumoniae, a chief cause of nosocomial pneumonia, is a versatile and commonly multidrug-resistant human pathogen for which further insight into pathogenesis is needed. We show that the pilus regulatory gene fimK promotes the virulence of K. pneumoniae strain TOP52 in murine pneumonia. This contrasts with the attenuating effect of fimK on urinary tract virulence, illustrating that a single factor may exert opposing effects on pathogenesis in distinct host niches. Loss of fimK in TOP52 pneumonia was associated with diminished lung bacterial burden, limited innate responses within the lung, and improved host survival. FimK expression was shown to promote serum resistance, capsule production, and protection from phagocytosis by host immune cells. Finally, while the widely used K. pneumoniae model strain 43816 produces rapid dissemination and death in mice, TOP52 caused largely localized pneumonia with limited lethality, thereby providing an alternative tool for studying K. pneumoniae pathogenesis and control within the lung.

  10. Bacteremic pneumonia caused by extensively drug-resistant Streptococcus pneumoniae.

    Science.gov (United States)

    Kang, Cheol-In; Baek, Jin Yang; Jeon, Kyeongman; Kim, So Hyun; Chung, Doo Ryeon; Peck, Kyong Ran; Lee, Nam Yong; Song, Jae-Hoon

    2012-12-01

    The emergence of antimicrobial resistance threatens the successful treatment of pneumococcal infections. Here we report a case of bacteremic pneumonia caused by an extremely drug-resistant strain of Streptococcus pneumoniae, nonsusceptible to at least one agent in all classes but vancomycin and linezolid, posing an important new public health threat in our region.

  11. Interleukin 6, lipopolysaccharide-binding protein and interleukin 10 in the prediction of risk and etiologic patterns in patients with community-acquired pneumonia: results from the German competence network CAPNETZ

    Directory of Open Access Journals (Sweden)

    Zobel Katrin

    2012-02-01

    Full Text Available Abstract Background The aim of our study was to investigate the predictive value of the biomarkers interleukin 6 (IL-6, interleukin 10 (IL-10 and lipopolysaccharide-binding protein (LBP compared with clinical CRB and CRB-65 severity scores in patients with community-acquired pneumonia (CAP. Methods Samples and data were obtained from patients enrolled into the German CAPNETZ study group. Samples (blood, sputum and urine were collected within 24 h of first presentation and inclusion in the CAPNETZ study, and CRB and CRB-65 scores were determined for all patients at the time of enrollment. The combined end point representative of a severe course of CAP was defined as mechanical ventilation, intensive care unit treatment and/or death within 30 days. Overall, a total of 1,000 patients were enrolled in the study. A severe course of CAP was observed in 105 (10.5% patients. Results The highest IL-6, IL-10 and LBP concentrations were found in patients with CRB-65 scores of 3-4 or CRB scores of 2-3. IL-6 and LBP levels on enrollment in the study were significantly higher for patients with a severe course of CAP than for those who did not have severe CAP. In receiver operating characteristic analyses, the area under the curve values for of IL-6 (0.689, IL-10 (0.665 and LPB (0.624 in a severe course of CAP were lower than that of CRB-65 (0.764 and similar to that of CRB (0.69. The accuracy of both CRB and CRB-65 was increased significantly by including IL-6 measurements. In addition, higher cytokine concentrations were found in patients with typical bacterial infections compared with patients with atypical or viral infections and those with infection of unknown etiology. LBP showed the highest discriminatory power with respect to the etiology of infection. Conclusions IL-6, IL-10 and LBP concentrations were increased in patients with a CRB-65 score of 3-4 and a severe course of CAP. The concentrations of IL-6 and IL-10 reflected the severity of disease in

  12. A thrombomodulin mutation that impairs active protein C generation is detrimental in severe pneumonia-derived gram-negative sepsis (melioidosis.

    Directory of Open Access Journals (Sweden)

    Liesbeth M Kager

    2014-04-01

    Full Text Available BACKGROUND: During severe (pneumosepsis inflammatory and coagulation pathways become activated as part of the host immune response. Thrombomodulin (TM is involved in a range of host defense mechanisms during infection and plays a pivotal role in activation of protein C (PC into active protein C (APC. APC has both anticoagulant and anti-inflammatory properties. In this study we investigated the effects of impaired TM-mediated APC generation during melioidosis, a common form of community-acquired Gram-negative (pneumosepsis in South-East Asia caused by Burkholderia (B. pseudomallei. METHODOLOGY/PRINCIPAL FINDINGS: (WT mice and mice with an impaired capacity to activate protein C due to a point mutation in their Thbd gene (TMpro/pro mice were intranasally infected with B. pseudomallei and sacrificed after 24, 48 or 72 hours for analyses. Additionally, survival studies were performed. When compared to WT mice, TMpro/pro mice displayed a worse survival upon infection with B. pseudomallei, accompanied by increased coagulation activation, enhanced lung neutrophil influx and bronchoalveolar inflammation at late time points, together with increased hepatocellular injury. The TMpro/pro mutation had limited if any impact on bacterial growth and dissemination. CONCLUSION/SIGNIFICANCE: TM-mediated protein C activation contributes to protective immunity after infection with B. pseudomallei. These results add to a better understanding of the regulation of the inflammatory and procoagulant response during severe Gram-negative (pneumosepsis.

  13. Detecção e classificação molecular de Chlamydophila psittaci em amostras fecais de aves assintomáticas

    Directory of Open Access Journals (Sweden)

    M.A. Braz

    2014-02-01

    Full Text Available Chlamydophila psittaci é uma bactéria que causa doença respiratória ou sistêmica em aves e em seres humanos. Em vista do risco de transmissão para humanos, o objetivo deste estudo foi detectar a presença de Chlamydophila spp. em amostras de fezes ou suabes cloacais de aves assintomáticas. Foram colhidas 403 amostras fecais ou suabes cloacais, provenientes de aves domésticas, selvagens ou exóticas. As amostras foram submetidas à PCR em tempo real para C. psittaci, para amplificação de fragmento parcial do gene da subunidade 16S do rRNA, utilizando o SsoFastTM EvaGreen® Supermix (Bio-Rad e análise da curva de dissociação. Para determinação do genótipo de C. psittaci, foi usada a hemi-nested PCR visando à amplificação de fragmento parcial do gene OMP-A, realizada nas amostras positivas pela PCR em tempo real, seguida de sequenciamento dos fragmentos amplificados. A PCR em tempo real revelou positividade em 17 (4,21% amostras. A hemi-nested foi positiva em 2 amostras positivas pela PCR em tempo real. O genótipo A de C. psittaci foi identificado pelo sequenciamento de uma amostra amplificada pela hemi-nested PCR. Os resultados deste experimento demonstram que a PCR em tempo real, visando à amplificação de fragmento parcial da subunidade 16S do rRNA, seguida da análise da curva de dissociação, pode ser utilizada para detecção de DNA de Chlamydophila sp. em amostras fecais de aves assintomáticas. A classificação da espécie de Chlamydophila e do genótipo de C. psittaci deve ser realizada por meio de PCR tendo como alvo o gene ompA e sequenciamento dos fragmentos amplificados.

  14. Enteral Tube Feeding and Pneumonia

    Science.gov (United States)

    Gray, David Sheridan; Kimmel, David

    2006-01-01

    To determine the effects of enteral tube feeding on the incidence of pneumonia, we performed a retrospective review of all clients at our institution who had gastrostomy or jejunostomy tubes placed over a 10-year period. Ninety-three subjects had a history of pneumonia before feeding tube insertion. Eighty had gastrostomy and 13, jejunostomy…

  15. Lymphocytic Interstitial Pneumonia.

    Science.gov (United States)

    Panchabhai, Tanmay S; Farver, Carol; Highland, Kristin B

    2016-09-01

    Lymphocytic interstitial pneumonia (LIP) is a rare lung disease on the spectrum of benign pulmonary lymphoproliferative disorders. LIP is frequently associated with connective tissue diseases or infections. Idiopathic LIP is rare; every attempt must be made to diagnose underlying conditions when LIP is diagnosed. Computed tomography of the chest in patients with LIP may reveal ground-glass opacities, centrilobular and subpleural nodules, and randomly distributed thin-walled cysts. Demonstrating polyclonality with immunohistochemistry is the key to differentiating LIP from lymphoma. The 5-year mortality remains between 33% and 50% and is likely to vary based on the underlying disease process.

  16. Caracterização epidemiológica e fatores de risco associados à infecção por Chlamydophila abortus em ovinos deslanados do semiárido brasileiro Epidemiological characterization and risk factors associated with Chlamydophila abortus infection in sheep in Brazilian semiarid

    Directory of Open Access Journals (Sweden)

    Areano E.M. Farias

    2013-03-01

    Full Text Available O objetivo do presente trabalho foi determinar as prevalências de propriedades positivas e animais soropositivos para Chlamydophila abortus em ovinos deslanados da região semiárida do Nordeste do Brasil, bem como identificar fatores de risco. Foram colhidas amostras de sangue de 476 ovinos procedentes de 72 propriedades em 14 municípios na mesorregião do Sertão, Estado da Paraíba. Para o diagnóstico sorológico da infecção por Chlamydophila abortus foi utilizada a reação de fixação de complemento (RFC. Uma propriedade foi considerada positiva quando apresentou pelo menos um animal soropositivo. Das 72 propriedades usadas 38 (52,8% apresentaram pelo menos um animal soropositivo, e dos 476 animais 94 (19,7% foram soropositivos. Participação em exposições (odds ratio =4,31; IC 95% =1,80-10,35; p=0,011 foi identificada como fator de risco. Sugere-se que a infecção por Chlamydophila abortus encontra-se disseminada em ovinos da região, e baseando-se na análise de fatores de risco, recomenda-se o controle sanitário nas exposições de animais.The aim of this investigation was to determine the flock-level and animal-level prevalences of Chlamydophila abortus in sheep from the semiarid region of Northeastern Brazil, as well as to identify risk factors. Blood samples were collected from 476 sheep of 72 flocks in 14 counties in the Sertão mesoregion, state of Paraíba. For the serological diagnosis of Chlamydophila abortus infection the complement fixation test (FC was carried out. A flock was positive when presented at least one seropositive animal. From the 72 flocks, 38 (52.8% presented at least one seropositive sheep, and 94 (19.7% of the 476 animals were seropositive. Participation in animal expositions (odds ratio= 4.31; 95% CI= 1.80-10.35; p=0.011 was identified as risk factor. It is suggested that C. abortus infection is spread in sheep of the region, and based on the risk factor analysis sanitary control in animal

  17. Protective contributions against invasive Streptococcus pneumoniae pneumonia of antibody and Th17-cell responses to nasopharyngeal colonisation.

    Science.gov (United States)

    Cohen, Jonathan M; Khandavilli, Suneeta; Camberlein, Emilie; Hyams, Catherine; Baxendale, Helen E; Brown, Jeremy S

    2011-01-01

    The nasopharyngeal commensal bacteria Streptococcus pneumoniae is also a frequent cause of serious infections. Nasopharyngeal colonisation with S. pneumoniae inhibits subsequent re-colonisation by inducing Th17-cell adaptive responses, whereas vaccination prevents invasive infections by inducing antibodies to S. pneumoniae capsular polysaccharides. In contrast, protection against invasive infection after nasopharyngeal colonisation with mutant S. pneumoniae strains was associated with antibody responses to protein antigens. The role of colonisation-induced Th17-cell responses during subsequent invasive infections is unknown. Using mouse models, we show that previous colonisation with S. pneumoniae protects against subsequent lethal pneumonia mainly by preventing bacteraemia with a more modest effect on local control of infection within the lung. Previous colonisation resulted in CD4-dependent increased levels of Th17-cell cytokines during subsequent infectious challenge. However, mice depleted of CD4 cells prior to challenge remained protected against bacteraemia, whereas no protection was seen in antibody deficient mice and similar protection could be achieved through passive transfer of serum. Serum from colonised mice but not antibody deficient mice promoted phagocytosis of S. pneumoniae, and previously colonised mice were able to rapidly clear S. pneumoniae from the blood after intravenous inoculation. Thus, despite priming for a Th17-cell response during subsequent infection, the protective effects of prior colonisation in this model was not dependent on CD4 cells but on rapid clearance of bacteria from the blood by antibody-mediated phagocytosis. These data suggest that whilst nasopharyngeal colonisation induces a range of immune responses, the effective protective responses depend upon the site of subsequent infection.

  18. The significance of myocardial enzyme,C-reactive protein and platelet count in children with bronchial pneumonia%血清心肌酶谱、C-反应蛋白和血小板计数变化在儿童支气管肺炎中的意义

    Institute of Scientific and Technical Information of China (English)

    王燕; 蒋昌科

    2014-01-01

    目的:探讨血清心肌酶谱、C-反应蛋白(CRP)和血小板计数与儿童支气管肺炎的关系。方法选择2010年8月至2012年12月该院确诊为支气管肺炎的1000例患儿为研究对象(支气管肺炎组),根据患儿的临床症状严重程度分为轻症肺炎组和重症肺炎组;并以1000例健康体检儿童为对照(对照组),分别测定其血液心肌酶谱、CRP和血小板的变化情况。结果支气管肺炎组心肌酶谱指标:天门冬氨酸氨基转移酶(AST)、乳酸脱氢酶(LDH)、a-羟丁酸脱氢酶(α-HBDH)、血清肌酸激酶(CK)、CK同工酶(CK-MB),血小板计数值和CRP水平均显著高于对照组,差异有统计学意义(P<0.05);而在重症肺炎组中,AST、LDH、α-HBDH、CK、CK-MB、血小板计数和CRP的升高比例亦显著显著高于轻症肺炎组,差异有统计学意义(P<0.05)。结论心肌酶谱、CRP和血小板联合检测对支气管肺炎的鉴别和诊断有较高特异度,尤其提高了对重症支气管肺炎鉴别的灵敏度。%Objective To explore the relationship between bronchial pneumonia in children and the level of myocardial enzyme , C-reactive protein (CRP) and platelet count .Methods 1 000 cases of children with bronchial pneumonia from August 2010 to De-cember 2010 were retrospectively analyzed (bronchial pneumonia group) ,According to the clinical symptom severity ,the children wwer divided into mild bronchial pneumonia group and severe bronchial pneumonia .1 000 healthy children were recruited in control group(control group) ,and the changes of myocardial enzyme ,platelet count and CRP were tested .Results The myocardial enzyme , including aminotransferase (AST ) ,lactate dehydrogenase (LDH ) ,a- hydroxybutyric acid dehydrogenase (alpha HBDH ) ,serum creatine kinase (CK) ,CK isoenzyme (CK-MB) ,platelet count and CRP in bronchial pneumonia group were all higher than that of control group (P<0 .05);In

  19. Proteomic analysis of the copper resistance of Streptococcus pneumoniae.

    Science.gov (United States)

    Guo, Zhong; Han, Junlong; Yang, Xiao-Yan; Cao, Kun; He, Ke; Du, Gaofei; Zeng, Guandi; Zhang, Liang; Yu, Guangchuang; Sun, Zhenghua; He, Qing-Yu; Sun, Xuesong

    2015-03-01

    Streptococcus pneumoniae is a Gram-positive bacterial pathogen causing a variety of diseases, including otitis media, bacteraemia and meningitis. Although copper is an essential trace metal for bacterial growth, high intracellular levels of free-copper are toxic. Copper resistance has emerged as an important virulence determinant of microbial pathogens. In this study, we determined the minimum inhibition concentration of copper for the growth inhibition of S. pneumoniae. Two-dimensional-electrophoresis coupled with mass spectrometry was applied to identify proteins involved in copper resistance of S. pneumoniae. In total, forty-four proteins with more than 1.5-fold alteration in expression (p < 0.05) were identified. Quantitative reverse transcription PCR was used to confirm the proteomic results. Bioinformatics analysis showed that the differentially expressed proteins were mainly involved in the cell wall biosynthesis, protein biosynthesis, purine biosynthesis, pyrimidine biosynthesis, primary metabolic process, and the nitrogen compound metabolic process. Many up-regulated proteins in response to the copper treatment directly or indirectly participated in the cell wall biosynthesis, indicating that the cell wall is a critical determinant in copper resistance of S. pneumoniae.

  20. Granzyme A impairs host defense during Streptococcus pneumoniae pneumonia.

    Science.gov (United States)

    van den Boogaard, Florry E; van Gisbergen, Klaas P J M; Vernooy, Juanita H; Medema, Jan P; Roelofs, Joris J T H; van Zoelen, Marieke A D; Endeman, Henrik; Biesma, Douwe H; Boon, Louis; Van't Veer, Cornelis; de Vos, Alex F; van der Poll, Tom

    2016-08-01

    Streptococcus pneumoniae is the most common causative pathogen in community-acquired pneumonia (CAP). Granzyme A (GzmA) is a serine protease produced by a variety of cell types involved in the immune response. We sought to determine the role of GzmA on the host response during pneumococcal pneumonia. GzmA was measured in bronchoalveolar lavage fluid (BALF) harvested from CAP patients from the infected and contralateral uninfected side and in lung tissue slides from CAP patients and controls. In CAP patients, GzmA levels were increased in BALF obtained from the infected lung. Human lungs showed constitutive GzmA expression by both parenchymal and nonparenchymal cells. In an experimental setting, pneumonia was induced in wild-type (WT) and GzmA-deficient (GzmA(-/-)) mice by intranasal inoculation of S. pneumoniae In separate experiments, WT and GzmA(-/-) mice were treated with natural killer (NK) cell depleting antibodies. Upon infection with S. pneumoniae, GzmA(-/-) mice showed a better survival and lower bacterial counts in BALF and distant body sites compared with WT mice. Although NK cells showed strong GzmA expression, NK cell depletion did not influence bacterial loads in either WT or GzmA(-/-) mice. These results implicate that GzmA plays an unfavorable role in host defense during pneumococcal pneumonia by a mechanism that does not depend on NK cells.

  1. Antimicrobial Resistant Streptococcus pneumoniae

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    B Khanal

    2010-09-01

    Full Text Available Introduction: Pneumococcal infections are important cause of morbidity and mortality. Knowledge of antimicrobial susceptibility patterns plays important role in the selection of appropriate therapy. Present study was undertaken to analyze the susceptibility patterns of pneumococcal isolates against commonly used antimicrobials with special reference to determination of minimum inhibitory concentration (MIC of penicillin in a tertiary care hospital in eastern Nepal. Methods: Twenty-six strains of S. pneumoniae isolated from various clinical specimens submitted to microbiology laboratory were evaluated. All isolates were tested for antimicrobial susceptibility by disk diffusion method. MIC of penicillin was tested by broth dilution method. Results: Of the total isolates 19 (73% were from invasive infections. Seven isolates were resistant to cotrimoxazole. No resistance to penicillin was seen in disk diffusion testing. Less susceptibility to penicillin (MIC 0.1-1.0 mg/L was observed in five (17% isolates. High level resistance to penicillin was not detected. One isolate was multidrug resistant. Conclusions: S. pneumoniaeisolates with intermediate resistance to penicillin prevail in Tertiary Care Hospital in eastern Nepal, causing invasive and noninvasive infections. As intermediate resistance is not detected in routine susceptibility testing, determination of MIC is important. It helps not only in the effective management of life threatening infections but is also essential in continuous monitoring and early detection of resistance. In addition, further study on pneumococcal infections, its antimicrobial resistance profile and correlation with clinical and epidemiological features including serotypes and group prevalence is recommended in future. Keywords: antimicrobial susceptibility pattern, penicillin, Streptococcus pneumoniae.

  2. Recombinant expression of Streptococcus pneumoniae capsular polysaccharides in Escherichia coli.

    Science.gov (United States)

    Kay, Emily J; Yates, Laura E; Terra, Vanessa S; Cuccui, Jon; Wren, Brendan W

    2016-04-01

    Currently, Streptococcus pneumoniae is responsible for over 14 million cases of pneumonia worldwide annually, and over 1 million deaths, the majority of them children. The major determinant for pathogenesis is a polysaccharide capsule that is variable and is used to distinguish strains based on their serotype. The capsule forms the basis of the pneumococcal polysaccharide vaccine (PPV23) that contains purified capsular polysaccharide from 23 serotypes, and the pneumococcal conjugate vaccine (PCV13), containing 13 common serotypes conjugated to CRM197 (mutant diphtheria toxin). Purified capsule from S. pneumoniae is required for pneumococcal conjugate vaccine production, and costs can be prohibitively high, limiting accessibility of the vaccine in low-income countries. In this study, we demonstrate the recombinant expression of the capsule-encoding locus from four different serotypes of S. pneumoniae within Escherichia coli. Furthermore, we attempt to identify the minimum set of genes necessary to reliably and efficiently express these capsules heterologously. These E. coli strains could be used to produce a supply of S. pneumoniae serotype-specific capsules without the need to culture pathogenic bacteria. Additionally, these strains could be applied to synthetic glycobiological applications: recombinant vaccine production using E. coli outer membrane vesicles or coupling to proteins using protein glycan coupling technology.

  3. Streptococcus pneumoniae serine protease HtrA, but not SFP or PrtA, is a major virulence factor in pneumonia.

    Science.gov (United States)

    de Stoppelaar, Sacha F; Bootsma, Hester J; Zomer, Aldert; Roelofs, Joris J T H; Hermans, Peter W M; van 't Veer, Cornelis; van der Poll, Tom

    2013-01-01

    Streptococcus (S.) pneumoniae is a common causative pathogen in pneumonia. Serine protease orthologs expressed by a variety of bacteria have been found of importance for virulence. Previous studies have identified two serine proteases in S. pneumoniae, HtrA (high-temperature requirement A) and PrtA (cell wall-associated serine protease A), that contributed to virulence in models of pneumonia and intraperitoneal infection respectively. We here sought to identify additional S. pneumoniae serine proteases and determine their role in virulence. The S. pneumoniae D39 genome contains five putative serine proteases, of which HtrA, Subtilase Family Protein (SFP) and PrtA were selected for insertional mutagenesis because they are predicted to be secreted and surface exposed. Mutant D39 strains lacking serine proteases were constructed by in-frame insertion deletion mutagenesis. Pneumonia was induced by intranasal infection of mice with wild-type or mutant D39. After high dose infection, only D39ΔhtrA showed reduced virulence, as reflected by strongly reduced bacterial loads, diminished dissemination and decreased lung inflammation. D39ΔprtA induced significantly less lung inflammation together with smaller infiltrated lung surface, but without influencing bacterial loads. After low dose infection, D39ΔhtrA again showed strongly reduced bacterial loads; notably, pneumococcal burdens were also modestly lower in lungs after infection with D39Δsfp. These data confirm the important role for HtrA in S. pneumoniae virulence. PrtA contributes to lung damage in high dose pneumonia; it does not however contribute to bacterial outgrowth in pneumococcal pneumonia. SFP may facilitate S. pneumoniae growth after low dose infection.

  4. J-UNIO protocol used for NMR structure determination of the 206-residue protein NP-346487.1 from Streptococcus pneumoniae TIGR4

    Energy Technology Data Exchange (ETDEWEB)

    Jaudzems, Kristaps [Latvian Institute of Organic Synthesis (Latvia); Pedrini, Bill [Paul Scherrer Institute (PSI), SwissFEL Project (Switzerland); Geralt, Michael; Serrano, Pedro; Wüthrich, Kurt, E-mail: wuthrich@scripps.edu [The Scripps Research Institute, Department of Integrative Structural and Computational Biology (United States)

    2015-01-15

    The NMR structure of the 206-residue protein NP-346487.1 was determined with the J-UNIO protocol, which includes extensive automation of the structure determination. With input from three APSY-NMR experiments, UNIO-MATCH automatically yielded 77 % of the backbone assignments, which were interactively validated and extended to 97 %. With an input of the near-complete backbone assignments and three 3D heteronuclear-resolved [{sup 1}H,{sup 1}H]-NOESY spectra, automated side chain assignment with UNIO-ATNOS/ASCAN resulted in 77 % of the expected assignments, which was extended interactively to about 90 %. Automated NOE assignment and structure calculation with UNIO-ATNOS/CANDID in combination with CYANA was used for the structure determination of this two-domain protein. The individual domains in the NMR structure coincide closely with the crystal structure, and the NMR studies further imply that the two domains undergo restricted hinge motions relative to each other in solution. NP-346487.1 is so far the largest polypeptide chain to which the J-UNIO structure determination protocol has successfully been applied.

  5. Ovarian hydrobursitis in female camels (Camelus dromedarius): the role of Chlamydophila abortus and a trial for medical treatment.

    Science.gov (United States)

    Ali, A; Al-Sobayil, F A; Hassanein, K M; Al-Hawas, A

    2012-06-01

    The occurrence of Chlamydophila abortus in female camels affected with ovarian hydrobursitis and a trial for medical treatment were studied. A total of 111 cases were included in two experiments. In Experiment 1, sera from 51 affected cases were tested for C. abortus antibody using enzyme-linked immunosorbent assay (ELISA). In Experiment 2, 60 female camels affected with bilateral ovarian hydrobursitis were divided into treated and control groups (n = 30 each). Based on the bursal diameter, females of both groups were subdivided into those having small ( 7 cm) bursae. Treated group received 20 mg/kg body weight oxytetracycline intramuscular, 4% lotagen intrauterine, and 500 μg cloprostenol intramuscular. Controls did not receive any treatment. All females were observed for 90 days non-return rate (NRR) and calving rate (CR). Antibodies against C. abortus were observed in 44/51 (86.3%) of the affected females. The 90 days NRR of the treated and control groups were 13/30 (43.3%) and 0/30 (0.0%), respectively, (P = 0.001), while the CR were 10/30 (33.3%) and 0/30 (0.0%), respectively, (P = 0.01). Based on bursal size, the 90 days NRR were 11/15 (73.3%), 2/7 (28.6%) and 0/8 (0.0%) for treated females having small, medium and large bursa, while the CR were 9/15 (60%), 1/7 (14.3%), and 0/8 (0.0%), respectively, (P = 0.01). In conclusion, it seems that C. abortus may be responsible for the spreading of the ovarian hydrobursitis syndrome in dromedaries. Small sized bursa could be medically treated.

  6. Genotyping of Chlamydophila psittaci using a new DNA microarray assay based on sequence analysis of ompA genes

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    Schubert Evelyn

    2008-04-01

    Full Text Available Abstract Background The currently used genotyping system for the avian zoonotic pathogen Chlamydophila (C. psittaci has evolved from serology and is based on ompA sequence variations. It includes seven avian and two non-avian genotypes. Restriction enzyme cleavage of the amplified ompA gene and, less frequently, ompA sequencing are being used for examination, but, beside methodological limitations, an increasing number of recently tested strains could not be assigned to any established genotype. Results Comprehensive analysis of all available ompA gene sequences has revealed a remarkable genetic diversity within the species C. psittaci, which is only partially covered by the present genotyping scheme. We suggest adjustments and extensions to the present scheme, which include the introduction of subgroups to the more heterogeneous genotypes A, E/B and D, as well as six provisional genotypes representing so far untypable strains. The findings of sequence analysis have been incorporated in the design of a new DNA microarray. The ArrayTube™ microarray-based ompA genotyping assay has been shown to discriminate among established genotypes and identify so far untyped strains. Its high specificity, which allows detection of single-nucleotide polymorphisms, is due to the parallel approach consisting in the use of 35 hybridization probes derived from variable domains 2 and 4 of the ompA gene. Conclusion The traditional genotyping system does not adequately reflect the extent of intra-species heterogeneity in ompA sequences of C. psittaci. The newly developed DNA microarray-based assay represents a promising diagnostic tool for tracing epidemiological chains, exploring the dissemination of genotypes and identifying non-typical representatives of C. psittaci.

  7. Prevalence of Chlamydophila psittaci infections in the eyes of cattle, buffaloes, sheep and goats in contact with a human population.

    Science.gov (United States)

    Osman, K M; Ali, H A; ElJakee, J A; Galal, H M

    2013-06-01

    This work is an example of cooperation between veterinary and human medicine being fully complementary and at the same time, indispensable to improve our knowledge on animal chlamydiosis. This study investigated the existence of ocular chlamydiae and determined the prevalence of its presence, chlamydiosis, in asymptomatic and diseased farm animals and adjacent humans. Data were obtained by the omp2 gene family Chlamydiaceae-specific PCR. Two hundred cattle, buffaloes, sheep and goats and 44 human specimens were also examined. Conjunctival swabs from both the eyes were collected from all animals and humans using cotton swabs. Samples were tested for chlamydiae by Vero cells tissue culture, chicken embryo, modified Gimenez staining, direct fluorescein-conjugated monoclonal antibody staining (FA), immunoperoxidase, CFT and PCR. The PCR-RFLP revealed that Chlamydophila psittaci demonstrated in the conjunctival samples of cattle (68% asymptomatic and 88% diseased), of buffalo (68% asymptomatic and 72% diseased), of sheep (68% asymptomatic and 80% diseased), of goat (76% asymptomatic and 92% diseased) and of humans (77% asymptomatic and 82% diseased). The Cp. psittaci was the only chlamydiae demonstrated in all of the ocular conjunctival samples, which confirms the prevalence of Cp. psittaci in this population of animals and adjacent humans. Statistically, the animal species factor was calculated and was found to be of no significance. Yet, there appeared to be a significant difference in the percentage of animal that tested positive using the different methods. Detection of Cp. psittaci in most samples confirms the prevalence of Cp. psittaci in this population of animals and adjacent humans.

  8. C-reactive protein as a tool of ventilator-associated pneumonia resolution%C反应蛋白检测在治疗呼吸机相关性肺炎中的作用

    Institute of Scientific and Technical Information of China (English)

    王泽宇; 吴允孚

    2013-01-01

    Objective To evaluate the role of C-reactive protein (CRP) , and to guide the antibiotic therapy in patients with ventilator-associated pneumonia resolution ( VAP). Methods Sixty patients with VAP in the department of ICU from Feb 2011 to Feb 2012 were randomly divided into the CRP group(30 patients) and the routine therapy group (30 patients). In the CRP group, the serum CRP levels were referenced to the quantity of antibiotic to administer; in the routine therapy group, antibiotics were administered according to regular treatment standers. Metrics compared between the two groups were the duration and cost of using antibiotic, overall duration of mechanical, the length of staying in ICU, the hospital mortality and the difference of inflammatory indexes(total white blood ceils, precent of neutrophil). Results The inflammatory indexes in the two groups all declined with no significant difference, the duration and cost of using antibiotic, o-verall duration of mechanical, the length of staying in ICU of PCT group were lower than those in the routine therapy group, the hospital mortality had no difference between the two groups. Conclusions CRP levels is important as a marker of ventilator-associated pneumonia resolution.%目的 评价C反应蛋白(C-reactive protein,CEP)对临床抗生素治疗呼吸机相关性肺炎(VAP)的应用价值. 方法 2011年2月至2012年2月在我院ICU入院VAP患者60例,随机分成CEP组(n=30)和常规组(n=30),常规治疗组按常规治疗方法使用抗生素进行治疗,CRP组通过血清CEP测定来指导抗生素使用,比较2组抗生素使用时间、机械通气时间、患者抗生素总费用、住院时间、病死率及抗生素治疗结束后2组患者炎性指标(白细胞总数,中性粒细胞百分比)之间的差异.结果 2组治疗后患者白细胞总数、中性粒细胞百分比差异无统计学意义,CRP组抗生素使用时间、机械通气时间,抗生素费用及住院时间均比常规组降

  9. Molecular epidemiology of penicillin-nonsusceptible Streptococcus pneumoniae among children in Greece

    NARCIS (Netherlands)

    D. Bogaert (Debby); G.A. Syrogiannopoulos; I.N. Grivea; R. de Groot (Ronald); N.G. Beratis; P.W.M. Hermans (Peter)

    2000-01-01

    textabstractA total of 145 penicillin-nonsusceptible Streptococcus pneumoniae strains were isolated from young carriers in Greece and analyzed by antibiotic susceptibility testing, serotyping, restriction fragment end labeling (RFEL), and penicillin-binding protein (PBP

  10. Molecular epidemiology of penicillin-nonsusceptible Streptococcus pneumoniae among children in Greece

    NARCIS (Netherlands)

    D. Bogaert (Debby); G.A. Syrogiannopoulos; I.N. Grivea; R. de Groot (Ronald); N.G. Beratis; P.W.M. Hermans (Peter)

    2000-01-01

    textabstractA total of 145 penicillin-nonsusceptible Streptococcus pneumoniae strains were isolated from young carriers in Greece and analyzed by antibiotic susceptibility testing, serotyping, restriction fragment end labeling (RFEL), and penicillin-binding protein

  11. Application of Whole Blood C-reactive Protein and White Blood Cell Count in Children With Pneumonia%全血C-反应蛋白与白细胞计数联合检测在小儿肺炎中的应用

    Institute of Scientific and Technical Information of China (English)

    温朝辉

    2016-01-01

    Objective To analyze the application of the detection of whole blood C-reactive protein and white blood cell count in infantile pneumonia.Methods 120 cases of children with pneumonia in our hospital from August 2013 to July 2015 were selected, there were 70 cases of bacterial bronchopneumonia and 50 cases of viral pneumonia among them. To selected another 60 healthy cases, to compare the whole blood C-reactive protein and white blood cell count about two groups.ResultsThe whole blood C-reactive protein and white blood cell count of the children with bacterial bronchopneumonia was higher than the children with viral pneumonia and the healthy cases, the difference was statistically significant (P<0.05). The whole blood C-reactive protein and white blood cell count of the children with the viral pneumonia was signiifcantly was higher than the healthy cases, the difference was statistically signiifcant (P<0.05).Conclusion The combination of whole blood C- reactive protein and white blood cell count can identify the type of pneumonia in children with pneumonia, and can detect and conifrm the bacterial bronchopneumonia faster.%目的:分析全血C-反应蛋白与白细胞计数联合检测在小儿肺炎中的应用。方法选取2013年8月~至2015年7月本院收治的肺炎患儿120例,其中细菌性支气管肺炎70例,病毒性肺炎50例,另再选取60例健康儿童,观察比较三组患儿的全血C-反应蛋白与白细胞计数。结果细菌性支气管肺炎患儿的全血C-反应蛋白和白细胞计数显著高于病毒性肺炎及健康儿童,差异具有统计学意义(P<0.05);病毒性肺炎患儿的全血C-反应蛋白与白细胞计数高于健康儿童,差异有统计学意义(P<0.05)。结论全血C-反应蛋白与白细胞计数联合检测在小儿肺炎中可以鉴别肺炎类型,能更快的检测及确诊出细菌性支气管肺炎。

  12. What Are the Signs and Symptoms of Pneumonia?

    Science.gov (United States)

    ... Home » Health Information for the Public » Health Topics » Pneumonia » Signs, Symptoms, and Complications Explore Pneumonia Pneumonia Causes Risk Factors Signs, Symptoms, and Complications ...

  13. [Idiopathic interstitial pneumonias in 2016].

    Science.gov (United States)

    Debray, M-P; Borie, R; Danel, C; Khalil, A; Majlath, M; Crestani, B

    2017-02-01

    Idiopathic interstitial pneumonias comprise 8 clinicopathological entities, most of them with a chronic course and various prognosis. Idiopathic pulmonary fibrosis is the most frequent and most severe of these. Computed tomography has an important role for its diagnosis. It can identify the corresponding pathological pattern of usual interstitial pneumonia in about 50 percent of cases. It can suggest differential diagnosis in other cases, most frequently fibrosing nonspecific interstitial pneumonia and chronic hypersensitivity pneumonitis. Imaging features should be integrated to clinical and available pathologic data during multidisciplinary team meetings involving physicians with a good knowledge of interstitial diseases. Some cases may be unclassifiable, but these could later be reclassified as new data may occur or imaging features may change. Surgical lung biopsy is being less frequently performed and an emerging less invasive technique, lung cryobiopsy, is under evaluation. Pleuroparenchymal fibroelastosis is a distinct entity only recently described, with uncertain prevalence and prognosis that seems being quite often associated to another pattern of interstitial pneumonia.

  14. CNS Complications of Mycoplasma Pneumoniae

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    J Gordon Millichap

    2004-08-01

    Full Text Available Three cases of acute central nervous system disease occurring subsequent to infection with M pneumoniae are reported from University College, Institute of Child Health, and Great Ormond Street Hospital, London, UK.

  15. [Pneumonia and its social representations].

    Science.gov (United States)

    Hamui-Sutton, Alicia; Nellen-Hummel, Haiko; Fernández-Ortega, Miguel Angel; Halabe-Cherem, José

    2009-01-01

    To correlate the sociostructural variables with the knowledge about pneumonia and to explore the social representations about the etiology, prevention, development and treatment in poor communities. A survey in 848 adults from seven Rural Health Centers affiliated to IMSS-Oportunidades Program in four States, was carried out. One-third of the sample did not understand the term pneumonia; 35 % of the patients with risk factors did not know its etiology; 43 % did not know about associated complications but 85 % considered that it causes death. The use of antibiotics was recognized as a therapeutic measure by 78 % and 20 % did not know how to prevent pneumonia. The findings showed a positive attitude to immunization but inadequate information about respiratory diseases. In neighborhoods with insufficient public services (purified water, electricity and paved roads) the ignorance about pneumonia tended to increase.

  16. Genotoxic Klebsiella pneumoniae in Taiwan.

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    Yi-Chyi Lai

    Full Text Available BACKGROUND: Colibactin is a nonribosomal peptide-polyketide synthesized by multi-enzyme complexes encoded by the pks gene cluster. Colibactin-producing Escherichia coli have been demonstrated to induce host DNA damage and promote colorectal cancer (CRC development. In Taiwan, the occurrence of pyogenic liver abscess (PLA has been suggested to correlate with an increasing risk of CRC, and Klebsiella pneumoniae is the predominant PLA pathogen in Taiwan. METHODOLOGY/PRINCIPAL FINDINGS: At the asn tRNA loci of the newly sequenced K. pneumoniae 1084 genome, we identified a 208-kb genomic island, KPHPI208, of which a module identical to the E. coli pks colibactin gene cluster was recognized. KPHPI208 consists of eight modules, including the colibactin module and the modules predicted to be involved in integration, conjugation, yersiniabactin production, microcin production, and unknown functions. Transient infection of BALB/c normal liver cells with K. pneumoniae 1084 increased the phosphorylation of histone H2AX, indicating the induction of host DNA damage. Colibactin was required for the genotoxicity of K. pneumoniae 1084, as it was diminished by deletion of clbA gene and restored to the wild type level by trans-complementation with a clbA coding plasmid. Besides, BALB/c mice infected with K. pneumoniae 1084 exhibited enhanced DNA damage in the liver parenchymal cells when compared to the isogenic clbA deletion mutant. By PCR detection, the prevalence of pks-positive K. pneumoniae in Taiwan is 25.6%, which is higher than that reported in Europe (3.5%, and is significantly correlated with K1 type, which predominantly accounted for PLA in Taiwan. CONCLUSIONS: Our knowledge regarding how bacteria contribute to carcinogenesis has just begun. The identification of genotoxic K. pneumoniae and its genetic components will facilitate future studies to elucidate the molecular basis underlying the link between K. pneumoniae, PLA, and CRC.

  17. Sinais clínicos e ocorrência de anticorpos anti-Chlamydophila abortus em ovinos de São Paulo e Minas Gerais Clinical signs and occurrence of antibodies anti-Chlamydophila abortus in ovines of São Paulo and Minas Gerais

    Directory of Open Access Journals (Sweden)

    Rodolfo Santos Rossi

    2012-11-01

    Full Text Available A Chlamydophila abortusi, anteriormente conhecida como Chlamydia psittaci sovovar 1, é uma bactéria Gram negativa, intracelular obrigatória. Esse micro-organismo é frequentemente encontrado em distúrbios reprodutivos em ovinos, bovinos e caprinos, sendo o aborto epizoótico dos bovinos e o aborto enzoótico dos ovinos e caprinos as manifestações mais importantes. Considerando-se o pouco material literário a respeito da clamidofilose no Brasil, a pesquisa teve como objetivo determinar a presença de anticorpos fixadores de complemento anti-Chlamydophila abortusi, correlacionando os resultados obtidos com achados no exame clínico e histórico dos animais, além de alterações nos índices zootécnicos, em especial na esfera reprodutiva, tais como alto índice de repetição de cio, número elevado de abortamentos, elevado número de natimortos, entre outros. Foram testadas para prova de fixação do complemento 220 amostras de soro de ovinos, de 26 propriedades, distribuídas em 19 municípios, com relato de manifestação reprodutiva, obtendo-se 19,55% (43/220 de testes positivos para Chlamydophila abortusi, com ocorrência de foco constatada de 61,53%. No geral, a titulação de anticorpos encontrada foi baixa, com título não superior a 64. A frequência de manifestação reprodutiva mais observada foi o aborto, representando 65,12% (28/43 do número total de animais soropositivos, seguido de repetição de cio juntamente com nascimento de cordeiro fraco, com frequência de 6,98% (3/43 e, por fim, morte neonatal com 4,65% (2/43, sendo que não houve associação significativa entre animais que foram positivos ao teste e a esses fatores.The Chlamydophila abortusi was previously known as Chlamydia psittaci sorovar 1, it is a Gram negative and obligate intracellular bacteria. This microorganism is frequently related with reproductive manifestation in ovines, goats and bovines. The major manifestation are Enzootic abortion in bovines and

  18. CRYSTAL STRUCTURE ANALYSIS OF A PUTATIVE OXIDOREDUCTASE FROM KLEBSIELLA PNEUMONIAE

    Energy Technology Data Exchange (ETDEWEB)

    Baig, M.; Brown, A.; Eswaramoorthy, S.; Swaminathan, S.

    2009-01-01

    Klebsiella pneumoniae, a gram-negative enteric bacterium, is found in nosocomial infections which are acquired during hospital stays for about 10% of hospital patients in the United States. The crystal structure of a putative oxidoreductase from K. pneumoniae has been determined. The structural information of this K. pneumoniae protein was used to understand its function. Crystals of the putative oxidoreductase enzyme were obtained by the sitting drop vapor diffusion method using Polyethylene glycol (PEG) 3350, Bis-Tris buffer, pH 5.5 as precipitant. These crystals were used to collect X-ray data at beam line X12C of the National Synchrotron Light Source (NSLS) at Brookhaven National Laboratory (BNL). The crystal structure was determined using the SHELX program and refi ned with CNS 1.1. This protein, which is involved in the catalysis of an oxidation-reduction (redox) reaction, has an alpha/beta structure. It utilizes nicotinamide adenine dinucleotide phosphate (NADP) or nicotine adenine dinucleotide (NAD) to perform its function. This structure could be used to determine the active and co-factor binding sites of the protein, information that could help pharmaceutical companies in drug design and in determining the protein’s relationship to disease treatment such as that for pneumonia and other related pathologies.

  19. The roentgenological study of measles pneumonia

    Energy Technology Data Exchange (ETDEWEB)

    Shin, U.; Song, C. H.; Lee, H. Y.; Chung, H. K.; Joo, K. B. [Han Gang Sacred Heart Hospital, Hallym College, Seoul (Korea, Republic of)

    1983-03-15

    Measles is important infectious disease of pediatrics and pneumonia is the most commonest complication of measles. We have experienced 20 cases of pneumonia among 31 cases of measles in infant nursing home of Chae Chun during of December. 1981. The results a are as follows; 1. The incidence of measles pneumonia is 64.5%. 2. The patterns of pneumonic infiltration is : The pneumonia may have a bronchopneumonia (60%), Lobar pneumonia (15%), or combined form (35%). 3. Both lungs are involved by measles pneumonia: Right lung only (30%), Left lung only (5%), or Bilateral (65%). 4. Hilar lymphadenopathy (51.6%). Hilar lymphadenopathy with pneumonia (82.2%) and hilar lymphadenopathy without pneumonia (17.8%). 5. There is no pulmonary nodule which is noted frequently in atypical measles pneumonia as a seguale.

  20. A Novel Metallo-β-Lactamase Involved in the Ampicillin Resistance of Streptococcus pneumoniae ATCC 49136 Strain.

    Directory of Open Access Journals (Sweden)

    Chia-Yu Chang

    Full Text Available Streptococcus pneumoniae, a penicillin-sensitive bacterium, is recognized as a major cause of pneumonia and is treated clinically with penicillin-based antibiotics. The rapid increase in resistance to penicillin and other antibiotics affects 450 million people globally and results in 4 million deaths every year. To unveil the mechanism of resistance of S. pneumoniae is thus an important issue to treat streptococcal disease that might consequently save millions of lives around the world. In this work, we isolated a streptococci-conserved L-ascorbate 6-phosphate lactonase, from S. pneumoniae ATCC 49136. This protein reveals a metallo-β-lactamase activity in vitro, which is able to deactivate an ampicillin-based antibiotic by hydrolyzing the amide bond of the β-lactam ring. The Michaelis parameter (Km = 25 μM and turnover number (kcat = 2 s-1 were obtained when nitrocefin was utilized as an optically measurable substrate. Through confocal images and western blot analyses with a specific antibody, the indigenous protein was recognized in S. pneumoniae ATCC 49136. The protein-overexpressed S. pneumonia exhibits a high ampicillin-tolerance ability in vivo. In contrast, the protein-knockout S. pneumonia reveals the ampicillin-sensitive feature relative to the wild type strain. Based on these results, we propose that this protein is a membrane-associated metallo-β-lactamase (MBL involved in the antibiotic-resistant property of S. pneumoniae.

  1. 肺炎衣原体、支原体在哮喘致病机制中的作用%Role of Chlamydophila pneumoniae and Mycoplasma pneumoniae in asthma pathogenesis

    Institute of Scientific and Technical Information of China (English)

    赵睿; 戚好文

    2007-01-01

    肺炎衣原体、支原体在哮喘致病机制中的作用引起了国际学者极大的兴趣.随着对肺炎衣原体、支原体与哮喘相关性认识程度的加深,结合众多学者所做的富有成效的研究成果,现将肺炎衣原体、支原体感染在哮喘致病机制中作用的研究进展作一综述.

  2. Evolutionary relationships among Chlamydophila abortus variant strains inferred by rRNA secondary structure-based phylogeny.

    Science.gov (United States)

    Siarkou, Victoria I; Stamatakis, Alexandros; Kappas, Ilias; Hadweh, Paul; Laroucau, Karine

    2011-01-01

    The evolutionary relationships among known Chlamydophila abortus variant strains including the LLG and POS, previously identified as being highly distinct, were investigated based on rRNA secondary structure information. PCR-amplified overlapping fragments of the 16S, 16S-23S intergenic spacer (IS), and 23S domain I rRNAs were subjected to cloning and sequencing. Secondary structure analysis revealed the presence of transitional single nucleotide variations (SNVs), two of which occurred in loops, while seven in stem regions that did not result in compensatory substitutions. Notably, only two SNVs, in 16S and 23S, occurred within evolutionary variable regions. Maximum likelihood and Bayesian phylogeny reconstructions revealed that C. abortus strains could be regarded as representing two distinct lineages, one including the "classical" C. abortus strains and the other the "LLG/POS variant", with the type strain B577(T) possibly representing an intermediate of the two lineages. The two C. abortus lineages shared three unique (apomorphic) characters in the 23S domain I and 16S-23S IS, but interestingly lacked synapomorphies in the 16S rRNA. The two lineages could be distinguished on the basis of eight positions; four of these comprised residues that appeared to be signature or unique for the "classical" lineage, while three were unique for the "LLG/POS variant". The U277 (E. coli numbering) signature character, corresponding to a highly conserved residue of the 16S molecule, and the unique G681 residue, conserved in a functionally strategic region also of 16S, are the most pronounced attributes (autapomorphies) of the "classical" and the "LLG/POS variant" lineages, respectively. Both lineages were found to be descendants of a common ancestor with the Prk/Daruma C. psittaci variant. Compared with the "classical", the "LLG/POS variant" lineage has retained more ancestral features. The current rRNA secondary structure-based analysis and phylogenetic inference reveal new

  3. Evolutionary relationships among Chlamydophila abortus variant strains inferred by rRNA secondary structure-based phylogeny.

    Directory of Open Access Journals (Sweden)

    Victoria I Siarkou

    Full Text Available The evolutionary relationships among known Chlamydophila abortus variant strains including the LLG and POS, previously identified as being highly distinct, were investigated based on rRNA secondary structure information. PCR-amplified overlapping fragments of the 16S, 16S-23S intergenic spacer (IS, and 23S domain I rRNAs were subjected to cloning and sequencing. Secondary structure analysis revealed the presence of transitional single nucleotide variations (SNVs, two of which occurred in loops, while seven in stem regions that did not result in compensatory substitutions. Notably, only two SNVs, in 16S and 23S, occurred within evolutionary variable regions. Maximum likelihood and Bayesian phylogeny reconstructions revealed that C. abortus strains could be regarded as representing two distinct lineages, one including the "classical" C. abortus strains and the other the "LLG/POS variant", with the type strain B577(T possibly representing an intermediate of the two lineages. The two C. abortus lineages shared three unique (apomorphic characters in the 23S domain I and 16S-23S IS, but interestingly lacked synapomorphies in the 16S rRNA. The two lineages could be distinguished on the basis of eight positions; four of these comprised residues that appeared to be signature or unique for the "classical" lineage, while three were unique for the "LLG/POS variant". The U277 (E. coli numbering signature character, corresponding to a highly conserved residue of the 16S molecule, and the unique G681 residue, conserved in a functionally strategic region also of 16S, are the most pronounced attributes (autapomorphies of the "classical" and the "LLG/POS variant" lineages, respectively. Both lineages were found to be descendants of a common ancestor with the Prk/Daruma C. psittaci variant. Compared with the "classical", the "LLG/POS variant" lineage has retained more ancestral features. The current rRNA secondary structure-based analysis and phylogenetic

  4. [Interstitial Pneumonia and Emphysema].

    Science.gov (United States)

    Sawa, Teiji; Kato, Yuko; Ishii, Sachiyo

    2015-09-01

    Interstitial pneumonia (IP) and chronic obstructive pulmonary disease (COPD) are representative diseases of restrictive pulmonary dysfunction and obstructive pulmonary dysfunction, respectively. In the preoperative anesthesia clinic, anesthesiologists are frequently asked to assess the anesthesia management of patients with these diseases. In respiratory function tests, IP is detected as a decrease in % vital capacity (< 80%), and COPD as a decrease in % FEV1.0 (< 70%). Other key factors which affect the assessment are; 1) severity assessment that affects the safety of anesthesia management, 2) prognostic evaluation including the acute exacerbation in the postoperative period, and 3) patient-related factors (age, life degree of autonomy, other comorbidities, surgery-related factors, and anesthesia method). In the patients in the disease stage I or II, anesthesia management is relatively safe. On the other hand, the patients in the disease stage IV have no surgical indication except life-saving emergent situation. In another words, anesthesiologists are required to make the judgment for the anesthesia management of the patient in the disease stage III, based on the assessment of patient-related factors, surgery-related factors, and prognosis.

  5. Endogenous tissue factor pathway inhibitor has a limited effect on host defence in murine pneumococcal pneumonia.

    Science.gov (United States)

    van den Boogaard, Florry E; van 't Veer, Cornelis; Roelofs, Joris J T H; Meijers, Joost C M; Schultz, Marcus J; Broze, George J; van der Poll, Tom

    2015-07-01

    Streptococcus (S.) pneumoniae is the most common causative pathogen in community-acquired pneumonia. Coagulation and inflammation interact in the host response to infection. Tissue factor pathway inhibitor (TFPI) is a natural anticoagulant protein that inhibits tissue factor (TF), the main activator of inflammation-induced coagulation. It was the objective of this study to investigate the effect of endogenous TFPI levels on coagulation, inflammation and bacterial growth during S. pneumoniae pneumonia in mice. The effect of low endogenous TFPI levels was studied by administration of a neutralising anti-TFPI antibody to wild-type mice, and by using genetically modified mice expressing low levels of TFPI, due to a genetic deletion of the first Kunitz domain of TFPI (TFPIK1(-/-)) rescued with a human TFPI transgene. Pneumonia was induced by intranasal inoculation with S. pneumoniae and samples were obtained at 6, 24 and 48 hours after infection. Anti-TFPI reduced TFPI activity by ~50 %. Homozygous lowTFPI mice and heterozygous controls had ~10 % and ~50 % of normal TFPI activity, respectively. TFPI levels did not influence bacterial growth or dissemination. Whereas lung pathology was unaffected in all groups, mice with ~10 % (but not with ~50 %) of TFPI levels displayed elevated lung cytokine and chemokine concentrations 24 hours after infection. None of the groups with low TFPI levels showed an altered procoagulant response in lungs or plasma during pneumonia. These data argue against an important role for endogenous TFPI in the antibacterial, inflammatory and procoagulant response during pneumococcal pneumonia.

  6. The Significance of Total IgE and C-reactive Protein of Children with Mycoplasma Pneumoniae Pneumonia%肺炎支原体肺炎患儿总IgE、C反应蛋白的变化及意义

    Institute of Scientific and Technical Information of China (English)

    刘建华

    2015-01-01

    Objective:To investigate the variation of total IgE and CRP of children with mycoplasma pneumoniae pneumonia in Shunyi District. Method:All subjects were selected in our hospital from Jan 2013 to Jan 2014,including MPP(n=80) and bacterial pneumonia(n=65) and healthy children(n=40) from January 2013 to January 2014.All children were drawed vein blood and analyzed MPP-IgM,total IgE and CRP.Result:Total IgE and CRP of MPP were higher than those of healthy children(P<0.001),and total IgE of MPP were higher than that of bacterial pneumonia(P<0.001).Conclusion:We should monitored CRP levels in early period of MPP.Enhancing immune function can improve MPP prognosis meanwhile regular anti-infection treatments.%目的:了解本地区肺炎支原体肺炎(mycoplasma pneumoniae pneumonia,MPP)患儿总IgE、C反应蛋白(C reactive protein,CRP)的水平变化,为临床治疗肺炎支原体肺炎提供指导。方法:选取2013年1月-2014年1月笔者所在医院收治的80例MPP患儿、65例细菌性肺炎患儿和40例健康体检的儿童,进行MPP抗体、总IgE和C反应蛋白测定,并进行比较分析。结果:MPP患儿总IgE、CRP高于健康体检儿童(P<0.001), MPP患儿总IgE高于细菌性肺炎患儿(P<0.001)。结论:MPP患儿早期应监测CRP变化,抗感染同时提高患儿免疫功能会改善患儿的预后。

  7. DAS181 Treatment of Severe Parainfluenza Virus 3 Pneumonia in Allogeneic Hematopoietic Stem Cell Transplant Recipients Requiring Mechanical Ventilation

    Directory of Open Access Journals (Sweden)

    B. Dhakal

    2016-01-01

    Full Text Available Parainfluenza virus (PIV may cause life-threatening pneumonia in allogeneic hematopoietic stem cell transplant (HSCT recipients. Currently, there are no proven effective therapies. We report the use of inhaled DAS181, a novel sialidase fusion protein, for treatment of PIV type 3 pneumonia in two allogeneic hematopoietic SCT recipients with respiratory failure.

  8. Impact of the factor V Leiden mutation on the outcome of pneumococcal pneumonia: a controlled laboratory study

    NARCIS (Netherlands)

    Schouten, M.; van 't Veer, C.; Roelofs, J.J.; Levi, M.; van der Poll, T.

    2010-01-01

    Introduction: Streptococcus (S.) pneumoniae is the most common cause of community-acquired pneumonia. The factor V Leiden (FVL) mutation results in resistance of activated FV to inactivation by activated protein C and thereby in a prothrombotic phenotype. Human heterozygous FVL carriers have been re

  9. [Fatal pneumonia caused by carbapenem resistant Klebsiella pneumoniae].

    Science.gov (United States)

    van Apeldoorn, Marjan; Lettinga, Kamilla; Bernards, Alexandra; Paltansing, Sunita; alNaiemi, Nashwan; Kalpoe, Jayant

    2010-01-01

    A 63-year-old Dutch man became colonized with a carbapenem resistant Klebsiella pneumoniae during a period of hospitalization in India. His recovery in the Netherlands was complicated by pneumonia due to this difficult-to-control multiresistant bacteria to which he eventually succumbed. Carbapenem resistance in Enterobacteriaceae, such as K. pneumoniae, is usually caused by carbapenemase (a betalactamase) production. Carbapenemase producing Enterobacteriaceae (CPE) are spreading throughout the world and cause difficult-to-treat infections that are associated with high mortality. This case report illustrates the clinical challenges associated with infection with these multiresistant Enterobacteriaceae. In the Netherlands, there are no guidelines for detection of CPE and carbapenemase production can frequently go undetected in clinical microbiology laboratories. As a consequence, adequate treatment of CPE infections and infection control measures to prevent the spread of CPE can be delayed. Expeditious development and implementation of existing Dutch draft guidelines for detection methods of CPE is therefore warranted.

  10. The clinical characteristics,treatment and outcome of macrolide-resistant Mycoplasma pneumoniae pneumonia in children

    Institute of Scientific and Technical Information of China (English)

    鲍芳

    2013-01-01

    Objective To investigate the drug resistance of My-coplasma pneumoniae among children with community-acquired pneumonia (CAP) ,and to explore the clinical and radiological characteristics of and the role of azithromycin in the treatment of of macrolide-resistant (MR) Mycoplasma pneumoniae pneumonia.Methods Cases of CAP in children (n=179) were prospectively enrolled in

  11. Investigation of the possible role of Chlamydophila abortus in reproductive failures in nrazilian herds of domestic ruminants

    Directory of Open Access Journals (Sweden)

    Francielle Gibson da Silva-Zacarias

    2009-11-01

    Full Text Available Chlamydophila abortus (C. abortus infection is related to reproductive failure in domestic ruminants. Although it has not been well characterized worldwide, this pathogen has already been identified in some European countries and in the USA. In Brazil, preliminary studies have shown serological evidence of C. abortus infection in herds with low antibody prevalence. Until now, the identification of C. abortus in biological samples from females presenting reproductive failures has not been described in Brazilian herds of domestic ruminants. The aim of this study was to evaluate the presence of the C. abortus in a collection of abortions from cattle (n=85, sheep (n=12, and goats (n=8, in samples of vaginal mucus from cows (n=13, sheep (n=90, and goats (n­=20, and in semen from sheep (n=10 and goats (n=5. The specimens (n=243 were evaluated using a PCR assay developed to amplify the 16S-23S rRNA intergenic space of C. abortus. A PCR assay with an internal control, which amplifies a fragment from the ND5 gene of bovine mitochondrial DNA, was used in order to evaluate the efficiency of the DNA extraction and of the PCR reaction. All biological samples (n=243 included in this study were negative for C. abortus in the PCR assay. The internal control enabled the amplification of a product from the bovine mitochondrial ND5 gene in all cattle abortion samples (n=85. Given the serological evidence indicating the presence of C. abortus infection in Brazilian herds of domestic ruminants, and considering the wide sampling evaluated, the failure to identify C. abortus in this survey suggests that the frequency of clinical signs in infected animals may be low or even absent.A infecção pela Chlamydophila abortus (C. abortus em ruminantes domésticos está relacionada com distúrbios reprodutivos. Apesar de ainda pouco estudada em todo o mundo, a infecção já foi identificada em alguns países europeus e também nos EUA. No Brasil, estudos preliminares

  12. Recent advances in the understanding of Chlamydophila pecorum infections, sixteen years after it was named as the fourth species of the Chlamydiaceae family.

    Science.gov (United States)

    Mohamad, Khalil Yousef; Rodolakis, Annie

    2010-01-01

    Chlamydophila pecorum found in the intestine and vaginal mucus of asymptomatic ruminants has also been associated with different pathological conditions in ruminants, swine and koalas. Some endangered species such as water buffalos and bandicoots have also been found to be infected by C. pecorum. The persistence of C. pecorum strains in the intestine and vaginal mucus of ruminants could cause long-term sub-clinical infection affecting the animal's health. C. pecorum strains present many genetic and antigenic variations, but coding tandem repeats have recently been found in some C. pecorum genes, allowing C. pecorum strains isolated from sick animals to be differentiated from those isolated from asymptomatic animals. This review provides an update on C. pecorum infections in different animal hosts and the implications for animal health. The taxonomy, typing and genetic aspects of C. pecorum are also reviewed.

  13. Kinase Activity Profiling of Pneumococcal Pneumonia

    NARCIS (Netherlands)

    Hoogendijk, Arie J.; Diks, Sander H.; van der Poll, Tom; Peppelenbosch, Maikel P.; Wieland, Catharina W.

    2011-01-01

    Background: Pneumonia represents a major health burden. Previous work demonstrated that although the induction of inflammation is important for adequate host defense against pneumonia, an inability to regulate the host's inflammatory response within the lung later during infection can be

  14. Bacterial pneumonia following acute ischemic stroke

    Directory of Open Access Journals (Sweden)

    Li-Fu Chen

    2013-02-01

    Conclusion: Pneumonia after acute ischemic stroke is a severe complication. Once stroke-related pneumonia develops, neurologic assessment, CURB-65 score, and shock can be used to predict the ultimate prognosis.

  15. Pneumonia Can Be Prevented -- Vaccines Can Help

    Science.gov (United States)

    ... What's this? Submit Button Past Emails CDC Features Pneumonia Can Be Prevented—Vaccines Can Help Language: English ... treatment (like antibiotics and antivirals). Save the Date: Pneumonia Twitter Chat on November 15 CDC experts will ...

  16. Symptoms, Diagnosis and Treatment of Pneumonia

    Science.gov (United States)

    ... Pneumonia Awareness Campaign, on World Pneumonia Day Blog: Yoga, Tai Chi and Your Lungs: The Benefits of ... number of items"); $("#local_list_xml").quickPagination(); }, error: function() { console.log("An error occurred while processing XML ...

  17. Outbreak of Pneumonia in the Setting of Fatal Pneumococcal Meningitis among US Army Trainees: Potential Role of Chlamydia pneumoniae Infection

    Science.gov (United States)

    2011-06-02

    physical stress may contribute to an increased risk for infections with Streptococcus pneumoniae , Streptococcus pyogenes, Mycoplasma pneumoniae ...Chlamydia pneumoniae , Mycoplasma pneumoniae , Streptococcus pneumoniae , Bordetella pertussis, and Legionella pneumophila[10] in addition to undergoing...postexposure chemoprophylaxis. Mil Med 2003;168:1-6 7. Balicer RD, Zarka S, Levine H, et al. Control of Streptococcus pneumoniae serotype 5 epidemic of

  18. Nursing home-acquired pneumonia.

    Science.gov (United States)

    El Solh, Ali A

    2009-02-01

    Nursing home-acquired pneumonia (NHAP) was first described in 1978. Since then there has been much written regarding NHAP and its management despite the lack of well-designed studies in this patient population. The most characteristic features of patients with NHAP are the atypical presentation, which may lead to delay in diagnosis and therapy. The microbial etiology of pneumonia encompasses a wide spectrum that spans microbes recovered from patients with community-acquired pneumonia to organisms considered specific only to nosocomial settings. Decision to transfer a nursing home patient to an acute care facility depends on a host of factors, which include the level of staffing available at the nursing home, patients' advance directives, and complexity of treatment. The presence of risk factors for multidrug-resistant pathogens dictates approach to therapy. Prevention remains the cornerstone of reducing the incidence of disease. Despite the advance in medical services, mortality from NHAP remains high.

  19. Cisplatin-Induced Eosinophilic Pneumonia

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    Hideharu Ideguchi

    2014-01-01

    Full Text Available A 67-year-old man suffering from esophageal cancer was admitted to our hospital complaining of dyspnea and hypoxemia. He had been treated with cisplatin, docetaxel, and fluorouracil combined with radiotherapy. Chest computed tomography revealed bilateral ground-glass opacity, and bronchoalveolar lavage fluid showed increased eosinophils. Two episodes of transient eosinophilia in peripheral blood were observed after serial administration of anticancer drugs before the admission, and drug-induced lymphocyte stimulation test to cisplatin was positive. Thus cisplatin-induced eosinophilic pneumonia was suspected, and corticosteroid was effectively administered. To our knowledge, this is the first reported case of cisplatin-induced eosinophilic pneumonia.

  20. Pneumonia acquired in the Community

    Directory of Open Access Journals (Sweden)

    María Caridad Fragoso Marchante

    2007-06-01

    Full Text Available A bibliographical revision of the main aspects in the diagnosis and treatment of the patients suffering from pneumonia acquired in the community is carried out. Microorganisms responsible for this type of pneumonia are mention in this paper as well as the available diagnostic methods for germs isolation. Different guidelines for diagnosis and treatment of this disease published by several medical societies and scientific institutions are analyzed by means of a review of the stratification index of the patients used in each of them. Aspects related to the duration of the treatment and the possible causes associated with the unfavorable evolution are stated.

  1. Enterobacter Asburiae Pneumonia with Cavitation

    Energy Technology Data Exchange (ETDEWEB)

    Cha, Seung Woo; Heo, Jeong Nam; Park, Choong Ki [Dept. of Radiology, Hanyang University College of Medicine, Guri Hospital, Guri (Korea, Republic of); Choi, Yo Won; Jeon, Seok Chol [Dept. of Radiology, Hanyang University College of Medicine, Seoul Hospital, Seoul (Korea, Republic of)

    2013-03-15

    Enterobacter species have increasingly been identified as pathogens over the past several decades. These bacterial species have become more important because most are resistant to cephalothin and cefoxitin, and can produce extended-spectrum {beta}-lactamase. Enterobacter asburiae (E. asburiae) is a gram-negative rod of the family Enterobacteriaceae, named in 1986. Since then, there has been only one clinical report of E. asburiae pneumonia. We report a case of E. asburiae pneumonia with cavitation and compare it with the previous case.

  2. Defective binding of the third component of complement (C3) to Streptococcus pneumoniae in multiple myeloma.

    Science.gov (United States)

    Cheson, B D; Walker, H S; Heath, M E; Gobel, R J; Janatova, J

    1984-04-01

    Patients with multiple myeloma (MM) are at an increased risk for infections with bacteria that require opsonization with complement. Because Streptococcus pneumoniae is the most frequently encountered pathogen in these patients, we investigated the ability of serum from patients with MM to mediate the binding of C3b, the major opsonin of the complement system, to S. pneumoniae. S. pneumoniae types 3, 14, and 25 were chosen for study, since S. pneumoniae type 3 activates primarily the classical complement pathway (CCP), type 25 primarily the alternative complement pathway (ACP), and type 14 both pathways. S. pneumoniae were treated with normal serum or serum from 17 patients with MM, and the bound C3b was quantified with fluorescein-conjugated anti-C3 in a spectrophotofluorometric assay. Despite normal or elevated serum concentrations of C3, total hemolytic complement, and C-reactive protein in all of the MM sera, factor B in 16/17 such sera, and C4 in 14/17 MM sera studied, all 17 sera demonstrated a defect in C3b binding to type 3 (32.7% +/- 6% of normal). In addition, serum from 15/17 patients bound decreased amounts of C3b to types 14 (39.6% +/- 8%) and 25 (52.2% +/- 8%). Mixing normal serum with MM serum restored MM C3b binding activity to all three S. pneumoniae types, suggesting that the defect was related to a deficiency rather than an inhibitor of C3 activation. Although MM patients are unable to produce specific antibodies to bacterial antigens, the addition of anti-S. pneumoniae antibodies to MM serum did not enhance C3b binding to any of the S. pneumoniae types. However, when S. pneumoniae were opsonized in a mixture of MM serum and C3-depleted normal serum, C3b binding was restored to all three S. pneumoniae types, demonstrating that MM C3 functions normally in the presence of other normal serum factors. In the present studies, the MM C3b binding defect appeared to correlate with the incidence of S. pneumoniae infections. Serum from patients with a

  3. A prospective study of sheep and goat abortion using real-time polymerase chain reaction and cut point estimation shows Coxiella burnetii and Chlamydophila abortus infection concurrently with other major pathogens.

    Science.gov (United States)

    Hazlett, Murray J; McDowall, Rebeccah; DeLay, Josepha; Stalker, Margaret; McEwen, Beverly; van Dreumel, Tony; Spinato, Maria; Binnington, Brian; Slavic, Durda; Carman, Susy; Cai, Hugh Y

    2013-05-01

    From 2009 to 2011, 163 sheep and 96 goat abortion submissions were received at the Animal Health Laboratory, University of Guelph, Ontario, Canada, for gross and histologic examination, as well as real-time polymerase chain reaction (PCR) testing for Chlamydophila abortus and/or Coxiella burnetii. Additional testing included immunohistochemistry for Toxoplasma gondii and Chlamydophila spp., routine bacterial culture and selective culture for Campylobacter spp., examination of modified acid-fast-stained placenta smears, enzyme-linked immunosorbent assay testing for Chlamydophila spp., and virus isolation. The final diagnosis made for each case by individual pathologists, based on gross and histologic lesions, as well as ancillary testing, was used as a standard to determine the significance of C. abortus and C. burnetii infection. Coxiella burnetii was identified by real-time PCR in 113 of 163 (69.0%) and 72 of 96 (75%) sheep and goat abortion submissions, respectively, but was considered to be significant in causing abortion in only 11 of 113 (10%) sheep and 15 out of 72 (21%) goat submissions that tested positive. Chlamydophila abortus was identified by real-time PCR in 42 of 162 (26%) and 54 of 92 (59%) sheep and goat submissions, respectively, but was considered the cause of the abortion in 16 of 42 (38%) sheep and 34 of 54 (63%) goat submissions that tested positive. Optimal sensitivity and specificity cut points for the real-time PCR copy number for C. abortus and C. burnetii were determined using the final pathology diagnosis as the reference test.

  4. Seropositivity for Chlamydia Pneumoniae and Mycoplasma Pneumoniae in Asthmatic Children

    Directory of Open Access Journals (Sweden)

    Murat Tutanc

    2014-03-01

    Full Text Available Acute respiratory tract infections may trigger acute asthma attacks and may be held responsible for etiopathogenesis in children with asthma. Although bacterial infections attract a limited amount of attention, recently Chlamydia pneumoniae (CP and Mycoplasma pneumoniae (MP, in particular, are reported to be the possible factors. IgM and IgG seroprevalence was investigated in 66 children patients with bronchial asthma (between the ages of 3 and 14 for CP and Mycoplasma pneumoniae. In a total of 66 cases, 18 (27.2% patients were detected with IgG positivity for CP whereas 27 of them (40.9% were detected with IgG positivity for MP. IgG positivity was determined in 6 patients (13.0% in the control group for CP, and in 6 patients (10.8% in the control group for MP. The rate of the asthma patients with IgG seropositivity for MP was 4 times higher than that of the control group. It was seen that IgG antibody seropositivity for CP was higher in those with more frequent attacks. No such difference was observed in terms of IgG antibody seropositivity for M. pneumoniae. There are many studies indicating that CP and MP infections take an importance place in the etiology of bronchial asthma and asthma attacks in children. The results obtained reveal the effect of both microorganisms on the etiopathogenesis of the bronchial asthma and the increased number of asthma attacks.

  5. Determination of penicillin susceptibility of Streptococcus pneumoniae using the polymerase chain reaction.

    OpenAIRE

    Jalal, H; Organji, S.; Reynolds, J.; Bennett, D; O'Mason, E.; Millar, M R

    1997-01-01

    AIM: To develop a polymerase chain reaction (PCR) based method to detect penicillin susceptibility in isolates of Streptococcus pneumoniae (SP). METHOD: PCR primers were designed to amplify differential nucleotide sequences of the penicillin-binding protein (PBP) genes 2b, 2x, and 1a in penicillin susceptible and resistant strains of SP. Primers derived from the PBP 2x and 2b genes were designed to amplify products from penicillin susceptible S pneumoniae (PSSP), whereas primers derived from ...

  6. Caracterização epidemiológica e fatores de risco associados à infecção por Chlamydophila abortus em ovinos deslanados do semiárido brasileiro

    Directory of Open Access Journals (Sweden)

    Areano E.M. Farias

    2013-03-01

    Full Text Available O objetivo do presente trabalho foi determinar as prevalências de propriedades positivas e animais soropositivos para Chlamydophila abortus em ovinos deslanados da região semiárida do Nordeste do Brasil, bem como identificar fatores de risco. Foram colhidas amostras de sangue de 476 ovinos procedentes de 72 propriedades em 14 municípios na mesorregião do Sertão, Estado da Paraíba. Para o diagnóstico sorológico da infecção por Chlamydophila abortus foi utilizada a reação de fixação de complemento (RFC. Uma propriedade foi considerada positiva quando apresentou pelo menos um animal soropositivo. Das 72 propriedades usadas 38 (52,8% apresentaram pelo menos um animal soropositivo, e dos 476 animais 94 (19,7% foram soropositivos. Participação em exposições (odds ratio =4,31; IC 95% =1,80-10,35; p=0,011 foi identificada como fator de risco. Sugere-se que a infecção por Chlamydophila abortus encontra-se disseminada em ovinos da região, e baseando-se na análise de fatores de risco, recomenda-se o controle sanitário nas exposições de animais.

  7. 肠脂肪酸结合蛋白在肺炎合并胃肠功能损伤患儿血清中的变化及意义%Change in serum intestinal fatty acid binding protein and its significance in children with pneumonia and gastrointestinal injury

    Institute of Scientific and Technical Information of China (English)

    范晓蕾; 李海英; 陈晓昕; 谢垒; 王怀立

    2016-01-01

    目的:探讨肠脂肪酸结合蛋白(IFABP)在社区获得性肺炎患儿血清中的变化及与胃肠功能损伤的相关性。方法选择2015年1月至10月的社区获得性肺炎患儿82例(轻症34例,重症48例),根据小儿危重病例评分(PCIS)将重症肺炎患儿分为非危重组(25例)和危重组(23例);另选取体检的健康儿童30例作为对照组。采用酶联免疫吸附法检测血清 IFABP 浓度,并对重症肺炎患儿进行急性胃肠损伤(AGI)分级。比较各组间血清 IFABP 浓度的差异,并对 IFABP 与 AGI 分级和 PCIS 进行相关性分析。结果重症肺炎血清 IFABP 浓度均高于对照组和轻症肺炎组(均 P<0.01),轻度肺炎组血清 IFABP 浓度亦明显高于对照组(P<0.01)。危重组血清 IFABP 浓度高于非危重组(P<0.01)。AGI 1~4级组血清 IFABP 均高于对照组(P<0.01),而且随着 AGI 级别的增高,血清 IFABP 浓度也逐渐增高,差异有统计学意义(P<0.01)。IFABP 与 AGI 分级呈正相关(P<0.01);与 PCIS 呈负相关(P<0.01)。结论肺炎患儿血清 IFABP 均有所增高,血清 IFABP 可以作为肺炎患儿合并胃肠损伤早期诊断和病情评估的敏感指标。%Objective To study the change in serum intestinal fatty acid binding protein (IFABP) in children with pneumonia and its correlation with gastrointestinal injury. Methods A total of 82 children with community-acquired pneumonia who were treated from January to October, 2015 were enrolled, among whom 34 had mild pneumonia and 48 had severe pneumonia. According to pediatric critical illness score (PCIS), the children with severe pneumonia were further divided into non-critical group (25 patients) and critical group (23 patients). Thirty healthy children who underwent physical examination at outpatient service were enrolled as the control group. ELISA was used to measure serum IFABP level, and the acute gastrointestinal injury (AGI

  8. Bacterial Pneumonia in Older Adults.

    Science.gov (United States)

    Marrie, Thomas J; File, Thomas M

    2016-08-01

    Community-acquired pneumonia is common in the elderly person; its presentation in this population is often confounded by multiple comorbid illnesses, including those that result in confusion. Although severity-of-illness scoring systems might aid decision-making, clinical judgment following a careful assessment is key in deciding on the site of care and appropriate therapy.

  9. Childhood Pneumonia Screener: a concept

    Directory of Open Access Journals (Sweden)

    Jukka Räsänen

    2014-06-01

    Full Text Available Childhood pneumonia continues to be the number one cause of death in children under five years of age in developing countries. In addition to mortality, pneumonia constitutes an enormous economic and social burden because late diagnosis is associated with high cost of treatment and often leads to chronic health problems. There are several bottlenecks in developing countries in the case flow of a child with lung infection: 1 recognising the symptoms as a reason to seek care, 2 getting the patient to a first-tier health facility, 3 scarcity of trained healthcare personnel who can diagnose the condition and its severity, 4 access to a second-tier facility in severe cases. These factors are commonly present in rural areas but even in more urban settings, access to a physician is often delayed. The Childhood Pneumonia Screener project aims at bridging the diagnostic gap using emerging technology. Mobile “smart” phone communication with several inexpensive dedicated sensors is proposed as a rapid data-collection and transmission unit that is connected to a central location where trained personnel assisted by sophisticated signal processing algorithms, evaluate the data and determine if the child is likely to have pneumonia and what the level and urgency of care should be.

  10. Clusters of Pneumocystis carinii pneumonia

    DEFF Research Database (Denmark)

    Helweg-Larsen, J; Tsolaki, A G; Miller, Raymonde

    1998-01-01

    types of P. carinii sp. f. hominis were identified in the samples from the patients with haematological malignancies, suggesting that this cluster of cases of P. carinii pneumonia was unlikely to have resulted from nosocomial transmission. A common ITS sequence type was observed in two of the patients...

  11. THE RELATION BETWEEN MINIMAL INHIBITORY CONCENTRATION OF PENICILLIN AND PENICILLIN-BINDING PRO-TEIN MUTATIONS OF STREPTOCOCCUS PNEUMONIAE%肺炎链球菌青霉素MIC与PBP变异的关系

    Institute of Scientific and Technical Information of China (English)

    黄卫青; 赵自云; 邓华

    2011-01-01

    Objective To explore the relationship between minimal inhibitory concentrations (MIC) of penicillin against streptococcus pneumoniae (Sp) and mutation of penicillin-binding proteins (PBP) 1A, 2B and 2X. Methods Fifty-one Sp strains were tested for penicillin sensitivity by MIC method, and DNA was extracted from each Sp strain, and PBP of PBP1A and PBP2B were amplified by polymerase chain reaction (PCR) followed by DNA sequencing. Amino acid sequences of PBP1A and PBD2B in conserved motifs were deduced by Clustalx software program. Results In all MIC≤1 mg/L of Sp, PBP1A was not found any mutation; while in MIC>1 mg/L, mutations appeared in most strains in several different sites, such as Thr574-(Asn), Ser575Thr, Gln576-Gly, and (Phe) 577-(Tyr) four consecutive residues,and the replacement of Thr371-Ser; any mutations of PBP2B were not found in PSSP when MIC≤1 mg/L; when MIC was greater than 1 mg/L and 2 mg/L, both 445Thr-Ala and Glu338-Gly appeared. Conclusion The mutation of PBP1A and PBP2B gene can lead to structural abnormalities of PBP1A-and-PBP2B-expressed PBP1A and PBP2B, decreasing the affinity with penicillin and other β-lactam antibiotics, and thus resistant to penicillin.%目的 探讨肺炎链球菌(Sp)青霉素抗生素最小抑菌质量浓度(MIC)与青霉素结合蛋白(PBP)1A、2B和2X变异的关系.方法 用MIC法对51株Sp进行青霉素药敏试验,并提取每株Sp的DNA,分别对其PBPIA、PBP2B中编码青霉素结合区域进行PCR扩增然后测序,并用软件进行序列比对,分析PBP1A、2B保守序列的氨基酸置换.结果 在所有MIC ≤1mg/L的青霉素敏感Sp(PSSP)中均未发现PBP1A突变,而在MIC>1mg/L时多数菌株出现Thr574-(Asn)、Ser575-Thr、Gln576-Gly、(Phe)577-(Tyr)4个连续残基的置换变异和Thr371-Ser 的置换.PBP2B 在所有MIC≤1 mg/L的PSSP中均未发生任何突变;而当MIC分别大于1 mg/L和2 mg/L时,均出现445Thr-Ala和G1u338-Gly.结论 PBP1A、PBP2B 基因突

  12. Complete Genome Sequence of Klebsiella pneumoniae YH43

    Science.gov (United States)

    Ogura, Yoshitoshi; Hayashi, Tetsuya; Mizunoe, Yoshimitsu

    2016-01-01

    We report here the complete genome sequence of Klebsiella pneumoniae strain YH43, isolated from sweet potato. The genome consists of a single circular chromosome of 5,520,319 bp in length. It carries 8 copies of rRNA operons, 86 tRNA genes, 5,154 protein-coding genes, and the nif gene cluster for nitrogen fixation. PMID:27081127

  13. Evaluation of Serum D-dimer Levels in Children with Pneumonia

    Directory of Open Access Journals (Sweden)

    Nilgün Selçuk Duru

    2016-03-01

    Full Text Available Aim: The aim of this study was to investigate the relationship of plasma D-dimer levels with duration of hospitalization and radiological and laboratory findings in patients with pneumonia. Methods: Forty-seven patients with pneumonia (31 boys and 16 girls, mean age: 4.2±4.7 years were included in the study. The patients were divided into two groups according to duration of hospitalization and three groups according to radiological findings. D-dimer and other laboratory findings were compared between the groups. Results: The mean serum D-dimer level was 1333.5±1364.4 ng/L. There was no statistically significant difference in D-dimer, leukocyte, erythrocyte sedimentation rate (ESR and C-reactive protein (CRP between the groups divided according to duration of hospitalization. In addition, there was no statistically difference in D-dimer levels between the groups divided according to radiological findings. Age, percentage of neutrophils, ESR and fibrinogen levels were higher in patients with lobar pneumonia when compared with the other groups and CRP level was higher in lobar pneumonia group when compared to interstitial pneumonia group. D-dimer levels were negatively correlated with age and positively correlated with ESR, CRP, and fibrinogen. Conclusion: In our study, D-dimer levels were high in patient with pneumonia. Further studies with a larger number of patients are necessary to determine the role of D-dimer levels as an acutephase reactant in patients with pneumonia

  14. Evaluation of different nucleic acid amplification techniques for the detection of M. pneumoniae, C. pneumoniae and Legionella spp. in respiratory specimens from patients with community-acquired pneumonia

    NARCIS (Netherlands)

    Loens, K; Beck, T; Ursi, D; Overdijk, M; Sillekens, P; Goossens, H; Ieven, M; Niesters, Bert

    2008-01-01

    The number of pathogens involved in community-acquired pneumonia, with varying susceptibilities to antimicrobials, is numerous constituting an enormous challenge for diagnostic microbiology. Differentiation of infections due to Streptococcus pneumoniae and those due to Mycoplasma pneumoniae, Chlamyd

  15. Phosphoproteomics of Klebsiella pneumoniae NTUH-K2044 Reveals a Tight Link between Tyrosine Phosphorylation and Virulence

    National Research Council Canada - National Science Library

    Miao-Hsia Lin; Tung-Li Hsu; Shu-Yu Lin; Yi-Jiun Pan; Jia-Tsrong Jan; Jin-Town Wang; Kay-Hooi Khoo; Shih-Hsiung Wu

    2009-01-01

    .... The capsular polysaccharide on K. pneumoniae surface was determined as the key to virulence. Although the regulation of capsular polysaccharide biosynthesis is largely unclear, it was found that protein-tyrosine kinases and phosphatases are involved...

  16. Mycoplasma pneumoniae associated organising pneumonia in a 10 year old boy.

    Science.gov (United States)

    Wachowski, O; Demirakça, S; Müller, K-M; Scheurlen, W

    2003-03-01

    We describe a 10 year old boy with organising pneumonia associated with acute Mycoplasma pneumoniae infection. The diagnosis of organising pneumonia was made by open lung biopsy and the M pneumoniae infection was proven serologically. Antibiotic and long term corticosteroid treatment resulted in steadily improving pulmonary function monitored by spirometry. The introduction of anti-inflammatory treatment with NSAIDs/immunosuppressive agents in order to spare steroids was well tolerated and resulted in further improvement of the pulmonary function. To our knowledge this is the first documented case of Mycoplasma pneumoniae associated organising pneumonia to be reported in a child.

  17. Toll-like receptor 4 agonistic antibody promotes innate immunity against severe pneumonia induced by coinfection with influenza virus and Streptococcus pneumoniae.

    Science.gov (United States)

    Tanaka, Akitaka; Nakamura, Shigeki; Seki, Masafumi; Fukudome, Kenji; Iwanaga, Naoki; Imamura, Yoshifumi; Miyazaki, Taiga; Izumikawa, Koichi; Kakeya, Hiroshi; Yanagihara, Katsunori; Kohno, Shigeru

    2013-07-01

    Coinfection with bacteria is a major cause of mortality during influenza epidemics. Recently, Toll-like receptor (TLR) agonists were shown to have immunomodulatory functions. In the present study, we investigated the effectiveness and mechanisms of the new TLR4 agonistic monoclonal antibody UT12 against secondary pneumococcal pneumonia induced by coinfection with influenza virus in a mouse model. Mice were intranasally inoculated with Streptococcus pneumoniae 2 days after influenza virus inoculation. UT12 was intraperitoneally administered 2 h before each inoculation. Survival rates were significantly increased and body weight loss was significantly decreased by UT12 administration. Additionally, the production of inflammatory mediators was significantly suppressed by the administration of UT12. In a histopathological study, pneumonia in UT12-treated mice was very mild compared to that in control mice. UT12 increased antimicrobial defense through the acceleration of macrophage recruitment into the lower respiratory tract induced by c-Jun N-terminal kinase (JNK) and nuclear factor kappaB (NF-κB) pathway-dependent monocyte chemoattractant protein 1 (MCP-1) production. Collectively, these findings indicate that UT12 promoted pulmonary innate immunity and may reduce the severity of severe pneumonia induced by coinfection with influenza virus and S. pneumoniae. This immunomodulatory effect of UT12 improves the prognosis of secondary pneumococcal pneumonia and makes UT12 an attractive candidate for treating severe infectious diseases.

  18. Clinical implications and treatment of multiresistant Streptococcus pneumoniae pneumonia.

    Science.gov (United States)

    File, T M

    2006-05-01

    Streptococcus pneumoniae is the leading bacterial cause of community-acquired respiratory tract infections. Prior to the 1970s this pathogen was uniformly susceptible to penicillin and most other antimicrobials. However, since the 1990s there has been a significant increase in drug-resistant Streptococcus pneumoniae (DRSP) due, in large part, to increased use of antimicrobials. The clinical significance of this resistance is not definitely established, but appears to be most relevant to specific MICs for specific antimicrobials. Certain beta-lactams (amoxicillin, cefotaxime, ceftriaxone), the respiratory fluoroquinolones, and telithromycin are among several agents that remain effective against DRSP. Continued surveillance studies, appropriate antimicrobial usage campaigns, stratification of patients based on known risk factors for resistance, and vaccination programmes are needed to appropriately manage DRSP and limit its spread.

  19. Community-onset Klebsiella pneumoniae pneumonia in Taiwan: clinical features of the disease and associated microbiological characteristics of isolates from pneumonia and nasopharynx.

    Science.gov (United States)

    Lin, Yi-Tsung; Wang, Yu-Ping; Wang, Fu-Der; Fung, Chang-Phone

    2015-01-01

    Klebsiella pneumoniae is an important cause of community-onset pneumonia in Asian countries and South Africa. We investigated the clinical characteristics of K. pneumoniae causing community-onset pneumonia, and the associated microbiological features between K. pneumoniae isolates from pneumonia and those from the nasopharynx in Taiwan. This study was conducted at the Taipei Veterans General Hospital during July, 2012 to February, 2014. The clinical characteristics in patients with community-onset K. pneumoniae pneumonia were analyzed. K. pneumoniae isolates from the nasopharynx of adults attending otorhinolaryngology outpatient clinics were collected to compare their microbiological features with those from pneumonia. Capsular genotypes, antimicrobial susceptibility, and multilocus sequence type (MLST) were determined among these strains. Ninety-one patients with community-onset K. pneumoniae pneumonia were enrolled. We found a high mortality (29.7%) among these patients. Capsular types K1, K2, K5, K20, K54, and K57 accounted for ∼70% of the K. pneumoniae isolates causing pneumonia, and ∼70% of all the K. pneumoniae strains isolated from the nasopharynx of patients in outpatient clinics. The MLST profiles further demonstrated the genetic relatedness between most pneumonia isolates and those from the nasopharynx. In conclusion, our results show that community-onset pneumonia caused by K. pneumoniae was associated with high mortality and could have a reservoir in the nasopharynx. To tackle this high-mortality disease, the distribution of capsular types in the nasopharynx might have implications for future vaccine development.

  20. Clinical case review: A method to improve identification of true clinical and radiographic pneumonia in children meeting the World Health Organization definition for pneumonia

    Directory of Open Access Journals (Sweden)

    Ruutu Petri

    2008-07-01

    Full Text Available Abstract Background The World Health Organization's (WHO case definition for childhood pneumonia, composed of simple clinical signs of cough, difficult breathing and fast breathing, is widely used in resource poor settings to guide management of acute respiratory infections. The definition is also commonly used as an entry criteria or endpoint in different intervention and disease burden studies. Methods A group of paediatricians conducted a retrospective review of clinical and laboratory data including C-reactive protein concentration and chest radiograph findings among Filipino children hospitalised in the Bohol Regional Hospital who were enrolled in a pneumococcal vaccine efficacy study and had an episode of respiratory disease fulfilling the WHO case definition for clinical pneumonia. Our aim was to evaluate which disease entities the WHO definition actually captures and what is the probable aetiology of respiratory infections among these episodes diagnosed in this population. Results Among the 12,194 children enrolled to the vaccine study we recorded 1,195 disease episodes leading to hospitalisation which fulfilled the WHO criteria for pneumonia. In total, 34% of these episodes showed radiographic evidence of pneumonia and 11% were classified as definitive or probable bacterial pneumonia. Over 95% of episodes of WHO-defined severe pneumonia (with chest indrawing had an acute lower respiratory infection as final diagnosis whereas 34% of those with non-severe clinical pneumonia had gastroenteritis or other non-respiratory infection as main cause of hospitalisation. Conclusion The WHO definition for severe pneumonia shows high specificity for acute lower respiratory infection and provides a tool to compare the total burden of lower respiratory infections in different settings. Trial registration ISRCTN62323832

  1. Streptococcus pneumoniae Drugs Resistance in Acute Rhinosinusitis

    Directory of Open Access Journals (Sweden)

    Chong Jie Hao

    2016-03-01

    Full Text Available Background: Acute rhinosinusitis that usually caused by Streptococcus pneumoniae becomes the reason why patients seek for medical care. Drugs resistance in Streptococcus pneumoniae is increasing worldwide. This study was conducted to determine drugs resistance of Streptococcus pneumonia from acute rhinosinusitis in Dr. Hasan Sadikin General Hospital. Methods: A descriptive laboratory study was conducted in June–October 2014 at the Laboratory of Microbiology Faculty of Medicine Universitas Padjadjaran. The sample was taken using nasopharyngeal swabbing from 100 acute rhinosinusitis patients in Dr. Hasan Sadikin General Hospital and planted on tryptic soy agar containing 5% sheep blood and 5 μg/ml of gentamicin sulphate and then incubated in 5% CO2 incubator at 37°C for 24 hours. The identification of Streptococcus pneumonia was performed by optochin test. The susceptibility test against Streptococcus pneumoniae was done using disk diffusion method.The antibiotic disks were trimethoprim-sulfamethoxazole, oxacillin, levofloxacin, azithromycin, and doxycycline. Results: Out of 100 samples, 8 of them were tested positive for Streptococcus pneumoniae. Three of Streptococcus pneumoniae isolates died with unknown reason after it were stored at -80 .The drugs resistance test showed the resistance of Streptococcus pneumonia to oxacillin, azithromycin and trimethoprim were 6, whereas levofloxacin and doxycycline are 4. Conclusions: Streptococcus pneumonia drugs resistance in acute rhinosinusitis shows the resistance of Streptococcus pneumoniae to oxacillin, azithromycin and trimethoprim are 6, whereas the resistance to levofloxacin and doxycycline are 4.

  2. An evaluation of emerging vaccines for childhood pneumococcal pneumonia

    Directory of Open Access Journals (Sweden)

    Zhang Jian Shayne F

    2011-04-01

    Full Text Available Abstract Background Pneumonia is the leading cause of child mortality worldwide. Streptococcus pneumoniae (SP or pneumococcus is estimated to cause 821,000 child deaths each year. It has over 90 serotypes, of which 7 to 13 serotypes are included in current formulations of pneumococcal conjugate vaccines that are efficacious in young children. To further reduce the burden from SP pneumonia, a vaccine is required that could protect children from a greater diversity of serotypes. Two different types of vaccines against pneumococcal pneumonia are currently at varying stages of development: a multivalent pneumococcal conjugate vaccine covering additional SP serotypes; and a conserved common pneumococcal protein antigen (PPA vaccine offering protection for all serotypes. Methods We used a modified CHNRI methodology for setting priorities in health research investments. This was done in two stages. In Stage I, we systematically reviewed the literature related to emerging SP vaccines relevant to several criteria of interest: answerability; efficacy and effectiveness; cost of development, production and implementation; deliverability, affordability and sustainability; maximum potential for disease burden reduction; acceptability to the end users and health workers; and effect on equity. In Stage II, we conducted an expert opinion exercise by inviting 20 experts (leading basic scientists, international public health researchers, international policy makers and representatives of pharmaceutical companies. The policy makers and industry representatives accepted our invitation on the condition of anonymity, due to sensitive nature of their involvement in such exercises. They answered questions from CHNRI framework and their “collective optimism” towards each criterion was documented on a scale from 0 to 100%. Results The experts expressed very high level of optimism (over 80% that low-cost polysaccharide conjugate SP vaccines would satisfy each of the 9

  3. Mutations in Streptococcus pneumoniae penicillin-binding protein 2x: importance of the C-terminal penicillin-binding protein and serine/threonine kinase-associated domains for beta-lactam binding.

    Science.gov (United States)

    Maurer, Patrick; Todorova, Katya; Sauerbier, Julia; Hakenbeck, Regine

    2012-06-01

    Penicillin-binding protein 2x (PBP2x) mutations that occur during the selection with beta-lactams are located within the central penicillin-binding/transpeptidase (TP) domain, and are believed to mediate resistance by interfering with the formation of a covalent complex of the active site serine with the antibiotic. We now investigated the effect of two point mutations found in two independently obtained laboratory mutants that are located at the surface of the TP domain with their side chains facing outside (G422D respectively R426C). They have no significant effect on resistance to cefotaxime in vivo or on binding to Bocillin™FL to the active site in vitro using purified PBP2x derivatives, thus apparently do not affect the active site directly. In contrast, in silico modeling revealed that they affect van der Waal's interactions with the PASTA1 (PBP and serine/threonine kinase associated) domain of the C-terminal extension and a noncovalent cefuroxime molecule found in the X-ray structure of an acylated PBP2x, suggesting some effect of the mutations on the interaction of the TP domain with PASTA1 and/or with the antibiotic associated with PASTA1. The effect of the PASTA domains on covalent binding of PBP2x to Bocillin FL was then investigated using a series of soluble truncated PBP2x derivatives. Deletion of 127 C-terminal residues, that is, of both PASTA domains, decreased binding dramatically by ∼90%. Surprisingly, deletion of only 40 amino acids resulted in the same phenotype, whereas the absence of 30 amino acids affected binding marginally by 10%, documenting a crucial role of the C-terminal domain for beta-lactam binding.

  4. Contribution of β-lactamases and porin proteins OmpK35 and OmpK36 to carbapenem resistance in clinical isolates of KPC-2-producing Klebsiella pneumoniae.

    Science.gov (United States)

    Zhang, Ying; Jiang, Xiaofei; Wang, Yanyan; Li, Gang; Tian, Yueru; Liu, Hong; Ai, Fuqi; Ma, Yiming; Wang, Bei; Ruan, Feiyi; Rajakumar, Kumar

    2014-01-01

    Fifty-seven carbapenem-resistant Klebsiella pneumoniae isolates belonging to ST11 (50 isolates), ST423 (5 isolates), and two other sequence types were studied. All were positive for blaKPC-2, blaTEM-1, and blaCTX-M-14. SDS-PAGE analysis of six representative isolates demonstrated varied porin expression. Nevertheless, when blaKPC-2 was deleted, carbapenem resistance was markedly reduced. Additionally, SHV-12, DHA-1, and/or VIM-1 appeared to contribute to accessory carbapenemase activity. In contrast, OmpK35 and/or OmpK36 deficiency seemed to serve only as a minor cooperative factor.

  5. Role of bacterial surface structures on the interaction of Klebsiella pneumoniae with phagocytes.

    Directory of Open Access Journals (Sweden)

    Catalina March

    Full Text Available Phagocytosis is a key process of the immune system. The human pathogen Klebsiella pneumoniae is a well known example of a pathogen highly resistant to phagocytosis. A wealth of evidence demonstrates that the capsule polysaccharide (CPS plays a crucial role in resistance to phagocytosis. The amoeba Dictyostelium discoideum shares with mammalian macrophages the ability to phagocytose and kill bacteria. The fact that K. pneumoniae is ubiquitous in nature and, therefore, should avoid predation by amoebae, poses the question whether K. pneumoniae employs similar means to counteract amoebae and mammalian phagocytes. Here we developed an assay to evaluate K. pneumoniae-D. discoideum interaction. The richness of the growth medium affected the threshold at which the cps mutant was permissive for Dictyostelium and only at lower nutrient concentrations the cps mutant was susceptible to predation by amoebae. Given the critical role of bacterial surface elements on host-pathogen interactions, we explored the possible contribution of the lipopolysaccharide (LPS and outer membrane proteins (OMPs to combat phagoyctosis by D. discoideum. We uncover that, in addition to the CPS, the LPS O-polysaccharide and the first core sugar participate in Klebsiella resistance to predation by D. discoideum. K. pneumoniae LPS lipid A decorations are also necessary to avoid predation by amoebae although PagP-dependent palmitoylation plays a more important role than the lipid A modification with aminoarabinose. Mutants lacking OMPs OmpA or OmpK36 were also permissive for D. discoideium growth. Except the LPS O-polysaccharide mutants, all mutants were more susceptible to phagocytosis by mouse alveolar macrophages. Finally, we found a correlation between virulence, using the pneumonia mouse model, and resistance to phagocytosis. Altogether, this work reveals novel K. pneumoniae determinants involved in resistance to phagocytosis and supports the notion that Dictyostelium amoebae

  6. [Ability of procalcitonin to predict bacteremia in patients with community acquired pneumonia].

    Science.gov (United States)

    Julián-Jiménez, Agustín; Timón Zapata, Jesús; Laserna Mendieta, Emilio José; Parejo Miguez, Raquel; Flores Chacartegui, Manuel; Gallardo Schall, Pablo

    2014-04-07

    To analyze the usefulness and ability of procalcitonin (PCT) to predict the presence of bacteremia in patients with community-acquired pneumonia (CAP) caused by Streptococcus pneumoniae (S. pneumoniae) or other bacteria. This is an observational, prospective and descriptive study involving patients who were diagnosed with CAP in our Emergency Department. Data collected included socio-demographic and comorbidity variables, Charlson index, stage in the Pneumonia Severity Index and criteria of severe NAC, microbiologic studies and biomarker determinations (PCT and C reactive protein). The follow-up was carried out during 30 days to calculate the predictive power and the diagnostic performance for bacteremia caused or not by S. pneumoniae. Four hundred and seventy-four patients were finally included in the study. Blood cultures were positive in 85 individuals (17.9%) and S. pneumoniae was identified as the responsible pathogen in 75 of them (88.4%) (in 5 cases together with another agent). The area under the Receiver Operating Characteristic curve for PCT to predict bacteremia (caused by S. pneumoniae or not) was 0.988 (95% confidence interval 0.908-0.995; P98% and>10, respectively. The most frequently isolated serotypes of S. pneumoniae were 19A, 7F, 1 and 3. The highest mean levels of PCT were found in serotypes 7F, 19A, 3 and 1, which showed statistically significant differences with regard to the others serotypes considered (P=.008). Serotypes associated with the highest percentage of severe sepsis-septic shock, 30-days mortality and multi-lobe or bilateral affection were 3, 1 and 19A; 1, 3 and 19A; and 3, 19A and 6A, respectively. PCT had a remarkable diagnostic ability to discard or suspect bacteremia and to guide the etiology of CAP caused by S. pneumoniae. Serotypes 1, 3, 19A and 7F showed greater frequency, systemic inflammatory response and clinical severity. Copyright © 2013 Elsevier España, S.L. All rights reserved.

  7. PERCH in Perspective: What Can It Teach Us About Pneumonia Etiology in Children?

    Science.gov (United States)

    Klugman, Keith P; Rodgers, Gail L

    2017-06-15

    The pneumonia team at the Bill & Melinda Gates Foundation congratulates the Pneumonia Etiology Research for Child Health (PERCH) study on delivering on their grant to collect high-quality data from thousands of children with World Health Organization-defined severe and very severe pneumonia and from controls in 9 diverse sites in 7 low- and middle-income countries. This supplement sets the foundation to understanding this complex study by providing an in-depth description of the study methodology, including discussion of key aspects such as antibiotic pretreatment, chest radiograph interpretation, utility of induced sputum in children, measurement of pathogen density, and use of C-reactive protein, and how these affect pneumonia etiology. © The Author 2017. Published by Oxford University Press for the Infectious Diseases Society of America.

  8. Protein

    Science.gov (United States)

    ... Food Service Resources Additional Resources About FAQ Contact Protein Protein is found throughout the body—in muscle, ... the heart and respiratory system, and death. All Protein Isn’t Alike Protein is built from building ...

  9. Persistent Pneumonia in an Infant.

    Science.gov (United States)

    Padilla, Kristen; Logan, Latania; Codispoti, Christopher; Jones, Carolyn; Van Opstal, Elizabeth

    2015-07-01

    A 4-month-old boy with past medical history of eczema presented with fever and cough; a chest radiograph showed lung consolidation, and he was initially treated with amoxicillin for presumed community-acquired pneumonia. After several days, his fever persisted. He was also profoundly anemic. Antibiotic coverage was broadened because of the concern for resistant organisms; he began to improve and was discharged from the hospital. However, at 5 months of age, his fever returned, and he continued to demonstrate lung consolidation on chest radiograph. Additionally, he had lost weight and continued to be anemic. Splenic cysts were noted on abdominal ultrasound. He was diagnosed with an unusual etiology for his pneumonia and improved with the appropriate therapy. An underlying immunodeficiency was suspected, but initial testing was nondiagnostic. At 12 months of age, he presented with another infection, and the final diagnosis was made.

  10. No causal association between inflammation and Chlamydia pneumoniae in patients with chronic ischemic arterial disease.

    Science.gov (United States)

    Altman, Raúl; Rouvier, Jorge; Scazziota, Alejandra; Gonzalez, Claudio

    2002-02-01

    The C-reactive protein, Chlamydia-specific IgG antibody, and fibrinogen were assayed in the serum of 159 patients with arterial disease (the arterial group) and 203 patients with heart valve prostheses (the valvular group) and no demonstrable coronary disease. In the arterial group, the Chlamydia pneumoniae antibody was > or = 1:32 for 67.3% (107/159) of the patients, the C-reactive protein was elevated in 41.5% (66/159), and the fibrinogen was elevated in 27.7% (44/159). In the valvular group, the C. pneumoniae antibody was > or = 1:32 for 59.1% (120/203) of the patients; the C-reactive protein was elevated in 34.0% (69/203), and the fibrinogen was elevated in 17.2% (35/203). Of 107 patients in the arterial group with C. pneumoniae titers > or = 1:32, only 26 (24.3%) had elevated fibrinogen (426 +/- 29 mg/dL) and 44 (41.1%) had elevated C-reactive protein (1.06 +/- 0.52 mg/dL). Similarly, of the 120 patients in the valvular group with C. pneumoniae titers > or = 1:32, 17 (14.2%) had elevated fibrinogen (409 +/- 29 mg/dL) and 34 had elevated C-reactive protein (0.99 +/- 1.1 mg/dL). Correlated poorly was C. pneumoniae with C-reactive protein and fibrinogen levels. Only the fibrinogen level could be discriminated between the arterial and the valvular group. These results suggest that no causal association exists between inflammation and C. pneumoniae. A highly significant correlation between C-reactive protein and fibrinogen levels was found.

  11. Biomarkers of Host Response Predict Primary End-Point Radiological Pneumonia in Tanzanian Children with Clinical Pneumonia: A Prospective Cohort Study.

    Directory of Open Access Journals (Sweden)

    Laura K Erdman

    Full Text Available Diagnosing pediatric pneumonia is challenging in low-resource settings. The World Health Organization (WHO has defined primary end-point radiological pneumonia for use in epidemiological and vaccine studies. However, radiography requires expertise and is often inaccessible. We hypothesized that plasma biomarkers of inflammation and endothelial activation may be useful surrogates for end-point pneumonia, and may provide insight into its biological significance.We studied children with WHO-defined clinical pneumonia (n = 155 within a prospective cohort of 1,005 consecutive febrile children presenting to Tanzanian outpatient clinics. Based on x-ray findings, participants were categorized as primary end-point pneumonia (n = 30, other infiltrates (n = 31, or normal chest x-ray (n = 94. Plasma levels of 7 host response biomarkers at presentation were measured by ELISA. Associations between biomarker levels and radiological findings were assessed by Kruskal-Wallis test and multivariable logistic regression. Biomarker ability to predict radiological findings was evaluated using receiver operating characteristic curve analysis and Classification and Regression Tree analysis.Compared to children with normal x-ray, children with end-point pneumonia had significantly higher C-reactive protein, procalcitonin and Chitinase 3-like-1, while those with other infiltrates had elevated procalcitonin and von Willebrand Factor and decreased soluble Tie-2 and endoglin. Clinical variables were not predictive of radiological findings. Classification and Regression Tree analysis generated multi-marker models with improved performance over single markers for discriminating between groups. A model based on C-reactive protein and Chitinase 3-like-1 discriminated between end-point pneumonia and non-end-point pneumonia with 93.3% sensitivity (95% confidence interval 76.5-98.8, 80.8% specificity (72.6-87.1, positive likelihood ratio 4.9 (3.4-7.1, negative likelihood ratio 0

  12. [Specificities of pneumonia in geriatrics].

    Science.gov (United States)

    Pepersack, T

    2014-09-01

    Pneumonia is a major cause of morbidity and mortality leading to a high rate of hospitalization especially in theelderly. It is often a sign of frailty and is associated with a poor prognosis. However, taking into account the geriatric specificities (risk factors, atypical clinical presentations with "geriatric syndromes", ethical debate) using an interdisciplinary and a comprehensive geriatric approach remains an important responsibility of the general practitionner. This article summarizes these specificities and offers interventions targeted on the characteristics of elderly patients.

  13. Severe community-acquired pneumonia caused by Mycoplasma pneumoniae in young female patient

    Directory of Open Access Journals (Sweden)

    Milačić Nena

    2015-07-01

    Full Text Available Mycoplasma pneumonia is common agent causing community acquired pneumonia in younger population. However, the course of illness is usually benign and is rarely associated with pulmonary complications. We report a 27 years old female patient with unilateral pneumonia followed by pleural effusion and adhesions on the same side. This potential source of infection should be considered in young patients where resolution of symptoms from pneumonia is delayed.

  14. Reflections on pneumonia in the tropics

    OpenAIRE

    Alpers, Michael P.

    2014-01-01

    This review of pneumonia in the tropics is based on experience with respiratory infections in Papua New Guinea since the 1970s. It discusses ideas, principles, historical aspects of pneumonia research and the need to work with people in the community. In order to understand pneumonia in a tropical setting and evaluate new interventions it is essential to study the ecosystem of the causative infections, within the host and the community and between interacting microorganisms. Vaccines are much...

  15. Heteroresistance to penicillin in Streptococcus pneumoniae.

    Science.gov (United States)

    Morand, Brigitte; Mühlemann, Kathrin

    2007-08-28

    Heteroresistance to beta-lactam antibiotics has been mainly described for staphylococci, for which it complicates diagnostic procedures and therapeutic success. This study investigated whether heteroresistance to penicillin exists in Streptococcus pneumoniae. Population analysis profile (PAP) showed the presence of subpopulations with higher penicillin resistance in four of nine clinical pneumococcal strains obtained from a local surveillance program (representing the multiresistant clones ST179, ST276, and ST344) and in seven of 16 reference strains (representing the international clones Spain(23F)-1, Spain(9V)-3, Spain(14)-5, Hungary(19A)-6, South Africa(19A)-13, Taiwan(23F)-15, and Finland(6B)-12). Heteroresistant strains had penicillin minimal inhibitory concentrations (MICs) (for the majority of cells) in the intermediate- to high-level range (0.19-2.0 mug/ml). PAP curves suggested the presence of subpopulations also for the highly penicillin-resistant strains Taiwan(19F)-14, Poland(23F)-16, CSR(19A)-11, and CSR(14)-10. PAP of bacterial subpopulations with higher penicillin resistance showed a shift toward higher penicillin-resistance levels, which reverted upon multiple passages on antibiotic-free media. Convergence to a homotypic resistance phenotype did not occur. Comparison of two strains of clone ST179 showed a correlation between the heteroresistant phenotype and a higher-penicillin MIC and a greater number of altered penicillin-binding proteins (PBP1a, -2b, and -2x), respectively. Therefore, heteroresistance to penicillin occurs in international multiresistant clones of S. pneumoniae. Pneumococci may use heteroresistance to penicillin as a tool during their evolution to high penicillin resistance, because it gives the bacteria an opportunity to explore growth in the presence of antibiotics before acquisition of resistance genes.

  16. Receptor for Advanced Glycation End Products (RAGE) Serves a Protective Role during Klebsiella pneumoniae - Induced Pneumonia.

    Science.gov (United States)

    Achouiti, Ahmed; de Vos, Alex F; van 't Veer, Cornelis; Florquin, Sandrine; Tanck, Michael W; Nawroth, Peter P; Bierhaus, Angelika; van der Poll, Tom; van Zoelen, Marieke A D

    2016-01-01

    Klebsiella species is the second most commonly isolated gram-negative organism in sepsis and a frequent causative pathogen in pneumonia. The receptor for advanced glycation end products (RAGE) is expressed on different cell types and plays a key role in diverse inflammatory responses. We here aimed to investigate the role of RAGE in the host response to Klebsiella (K.) pneumoniae pneumonia and intransally inoculated rage gene deficient (RAGE-/-) and normal wild-type (Wt) mice with K. pneumoniae. Klebsiella pneumonia resulted in an increased pulmonary expression of RAGE. Furthermore, the high-affinity RAGE ligand high mobility group box-1 was upregulated during K. pneumoniae pneumonia. RAGE deficiency impaired host defense as reflected by a worsened survival, increased bacterial outgrowth and dissemination in RAGE-/- mice. RAGE-/- neutrophils showed a diminished phagocytosing capacity of live K. pneumoniae in vitro. Relative to Wt mice, RAGE-/- mice demonstrated similar lung inflammation, and slightly elevated-if any-cytokine and chemokine levels and unchanged hepatocellular injury. In addition, RAGE-/- mice displayed an unaltered response to intranasally instilled Klebsiella lipopolysaccharide (LPS) with respect to pulmonary cell recruitment and local release of cytokines and chemokines. These data suggest that (endogenous) RAGE protects against K. pneumoniae pneumonia. Also, they demonstrate that RAGE contributes to an effective antibacterial defense during K. pneumoniae pneumonia, at least partly via its participation in the phagocytic properties of professional granulocytes. Additionally, our results indicate that RAGE is not essential for the induction of a local and systemic inflammatory response to either intact Klebsiella or Klebsiella LPS.

  17. Frequently Asked Questions about Ventilator-Associated Pneumonia

    Science.gov (United States)

    ... Submit Button Frequently Asked Questions about Ventilator-associated Pneumonia Recommend on Facebook Tweet Share Compartir What is a Ventilator-associated Pneumonia (VAP)? Ventilator-associated pneumonia (VAP) is a lung ...

  18. Klebsiella pneumoniae inoculants for enhancing plant growth

    Energy Technology Data Exchange (ETDEWEB)

    Triplett, Eric W. (Middleton, WI); Kaeppler, Shawn M. (Oregon, WI); Chelius, Marisa K. (Greeley, CO)

    2008-07-01

    A biological inoculant for enhancing the growth of plants is disclosed. The inoculant includes the bacterial strains Herbaspirillum seropedicae 2A, Pantoea agglomerans P101, Pantoea agglomerans P102, Klebsiella pneumoniae 342, Klebsiella pneumoniae zmvsy, Herbaspirillum seropedicae Z152, Gluconacetobacter diazotrophicus PA15, with or without a carrier. The inoculant also includes strains of the bacterium Pantoea agglomerans and K. pneumoniae which are able to enhance the growth of cereal grasses. Also disclosed are the novel bacterial strains Herbaspirillum seropedicae 2A, Pantoea agglomerans P101 and P102, and Klebsiella pneumoniae 342 and zmvsy.

  19. Klebsiella pneumoniae inoculants for enhancing plant growth

    Science.gov (United States)

    Triplett, Eric W [Middleton, WI; Kaeppler, Shawn M [Oregon, WI; Chelius, Marisa K [Greeley, CO

    2008-07-01

    A biological inoculant for enhancing the growth of plants is disclosed. The inoculant includes the bacterial strains Herbaspirillum seropedicae 2A, Pantoea agglomerans P101, Pantoea agglomerans P102, Klebsiella pneumoniae 342, Klebsiella pneumoniae zmvsy, Herbaspirillum seropedicae Z152, Gluconacetobacter diazotrophicus PA15, with or without a carrier. The inoculant also includes strains of the bacterium Pantoea agglomerans and K. pneumoniae which are able to enhance the growth of cereal grasses. Also disclosed are the novel bacterial strains Herbaspirillum seropedicae 2A, Pantoea agglomerans P101 and P102, and Klebsiella pneumoniae 342 and zmvsy.

  20. 青霉素结合蛋白PBP1a的克隆表达和对肺炎链球菌耐药性的影响%Cloning and expression of penicillin-binding protein PBP1a and its effect on the penicillin resistance of Streptococcus pneumonia

    Institute of Scientific and Technical Information of China (English)

    潘玲; 周侠; 杨致邦; 邓西川

    2012-01-01

    Objective To determine the effect of PBPla on the resistance of Streptococcus pneumonia to penicillin. The pbpla gene coding for a penicillin-binding protein in S. pneumoniae was cloned and expressed. Methods The pbpla fragment containing the STMK region from S. pneumoniae was chemically synthesized and amplified with PCR. The amplified products were identified by 1. 5% agarose electrophoretic analysis. The amplified pbpla was ligated with a pUC19 vector. The recombinant pUC19-pbpla plasmid was converted and translated into DH5α. The pbpla gene in DH5α was amplified and identified by 1. 5% agarose electrophoretic analysis again and then subjected to DNA sequencing analysis. Then the pUC19-pbpla was transformed into S. pneumoniae with sensitivity, intermediate sensitivity, and resistance to penicillin and with various mutations of pbpla and pbp2b. The expression of PBPla was analyzed by SDS-PAGE and the MIC of penicillin for the transformed S. pneumoniae was detected. Results A band of about 330 bp was observed in agarose electrophoretic analysis after pbpla was amplified with PCR, and this size coincided with expectations. The recombinant plasmid was 360 bp in length, and the sequences of the recombinant plasmid lacked mutations such as missing or inserted bases. The MIC of penicillin for strains with sensitivity and intermediate sensitivity to penicillin and with no pbpla mutation did not change. The MIC of penicillin for strains with intermediate sensitivity and pbpla mutation decreased slightly. The MIC for strains with resistance to penicillin and pbpla mutation decreased markedly. There were significant differences between transformed and untransformed S. pneumoniae(P<0. 01). Conclusion The penicillin-binding protein PBPla of S. pneumoniae was successfully expressed and markedly decreased the resistance of S. pneumoniae to penicillin.%目的 克隆并表达肺炎链球菌青霉素结合蛋白PBP1a,探讨其对青霉素耐药的影响. 方法 化学合

  1. [Pneumonia: The urgent problem of 21st century medicine].

    Science.gov (United States)

    Chuchalin, A G

    2016-01-01

    The paper analyzes the systematic reviews and meta-analyses on the strategic issues of pneumonia, which have been published in the past 3 years. It discusses the prevalence and mortality rates of acquired pneumonia, hospital-acquired (nosocomial) pneumonia, healthcare-associated pneumonia, ventilator-associated pneumonia, and Mycoplasma pneumonia, and the specific features of their etiology, diagnosis, and treatment. A large number of investigations emphasize the relevance of this problem in current clinical practice.

  2. Growth of the Pittsburgh Pneumonia Agent in Animal Cell Cultures

    OpenAIRE

    Rinaldo, Charles R.; Pasculle, A. William; Myerowitz, Richard L.; Gress, Francis M.; Dowling, John N.

    1981-01-01

    Pittsburgh pneumonia agent (Legionella micdadei) grew in monkey, chicken, and human cell cultures. Pittsburgh pneumonia agent grew predominantly in the cytoplasm, resulting in a nonfocal, mild cytopathic effect.

  3. The Type III Secretion System (T3SS of Chlamydophila psittaci Is Involved in the Host Inflammatory Response by Activating the JNK/ERK Signaling Pathway

    Directory of Open Access Journals (Sweden)

    Qing-zhi He

    2015-01-01

    Full Text Available Chlamydophila psittaci (C. psittaci is a human zoonotic pathogen, which could result in severe respiratory disease. In the present study, we investigated the role and mechanism of the type III secretion system (T3SS of C. psittaci in regulating the inflammatory response in host cells. C. psittaci-infected THP-1 cells were incubated with the specific T3SS inhibitor INP0007, inhibitors of ERK, p38, or JNK, and the levels of inflammatory cytokines were analyzed using Q-PCR and ELISA. The levels of ERK, p38, and JNK phosphorylation were analyzed by Western blot. Our results verified that INP0007 inhibited chlamydial growth in vitro, but the coaddition of exogenous iron completely reversed the growth deficit. INP0007 inhibited the growth of C. psittaci and decreased the levels of IL-8, IL-6, TNF-α, and IL-1β. Exogenous iron restored the chlamydial growth but not the production of inflammatory cytokines. These results demonstrated that the expression of inflammatory cytokines during infection was associated with the T3SS which reduced by incubation with ERK and JNK inhibitors, but not with p38 inhibitor. We concluded that the T3SS elicited inflammatory responses by activating the JNK or ERK signaling pathways in the infection of C. psittaci.

  4. Chlamydia trachomatis ompA genotypes in male patients with urethritis in Greece: conservation of the serovar distribution and evidence for mixed infections with Chlamydophila abortus.

    Science.gov (United States)

    Psarrakos, Panagiotis; Papadogeorgakis, Eleni; Sachse, Konrad; Vretou, Evangelia

    2011-08-01

    PCR amplification and nucleotide sequencing of the ompA gene of Chlamydia trachomatis were used to determine the prevalence and distribution of genotypes in 51 urine and urethral specimens from Greek male patients with urethritis, that were positive by the COBAS Amplicor test. A single C. trachomatis serovar was identified in 43 of the 51 amplified samples. Serovars F and E were the most prevalent (both 12, 28%), followed by D (9, 21%), G (4, 9%), B and K (both 2, 5%) and H and J (both 1, 2%). Over one third of the samples bared a variant ompA genotype that had been previously identified in other areas worldwide. Two results in this study, both observed for the first time, were of particular interest. First, the emergence of the unique variant genotype D/Ep6 (X77364.2) identified in 3 urethral samples. Second, the ompA genotype OCLH196 of the animal pathogen Chlamydophila abortus as well as a 23S rRNA gene fragment of this species detected by the assay ArrayTube™ was found in 7 urethral samples. The implications resulting from this observation for the health of the general population are discussed.

  5. Within-population diversity of koala Chlamydophila pecorum at ompA VD1-VD3 and the ORF663 hypothetical gene.

    Science.gov (United States)

    Higgins, D P; Beninati, T; Meek, M; Irish, J; Griffith, J E

    2012-05-04

    Infection of koalas by Chlamydophila pecorum is very common and causes significant morbidity, infertility and mortality. Fundamental to management of the disease is an understanding of the importance of multi-serotype infection or pathogen virulence in pathogenesis; these may need consideration in plans involving koala movement, vaccination, or disease risk assessment. Here we describe diversity of ompA VD1-3, and ORF663 hypothetical gene tandem repeat regions, in a single population of koalas with diverse disease outcomes. We PCR amplified and sequenced 72 partial ompA segments and amplified 25 tandem repeat segments (ORF663 hypothetical gene) from C. pecorum obtained from 62 koalas. Although several ompA genotypes were identified nationally, only one ompA genotype existed within the population studied, indicating that severe chlamydial disease occurs commonly in free-ranging koalas in the absence of infection by multiple MOMP serotypes of C. pecorum. In contrast, variation in tandem repeats within the ORF663 hypothetical gene was very high, approaching the entire range reported for pathogenic and non-pathogenic C. pecorum of European ruminants; providing an impetus for further investigation of this as a potential virulence trait.

  6. Soluble Endothelial Selectin in Acute Lung Injury Complicated by Severe Pneumonia

    Directory of Open Access Journals (Sweden)

    Daisuke Osaka, Yoko Shibata, Kazunori Kanouchi, Michiko Nishiwaki, Tomomi Kimura, Hiroyuki Kishi, Shuichi Abe, Sumito Inoue, Yoshikane Tokairin, Akira Igarashi, Keiko Yamauchi, Yasuko Aida, Takako Nemoto, Keiko Nunomiya, Koji Fukuzaki, Isao Kubota

    2011-01-01

    Full Text Available Background: Pneumonia is still one of the most frequent causes of death in the elderly. Complication of acute lung injury (ALI/acute respiratory distress syndrome (ARDS by pneumonia makes patients very ill due to severe respiratory failure. Biomarkers that can discriminate the presence of complicating ALI/ARDS are required for early detection. The aim of this research was to investigate whether soluble endothelial selectin (sES could be a biomarker for ALI.Methods: Serum sES levels were measured in 27 pneumonia patients, who were enrolled between April 2006 and September 2007. Among these patients, six had ALI or a condition that was clinically comparable to ALI (cALI. All patients who were enrolled were successfully treated and survived.Results: Circulating sES levels were elevated in pneumonia patients with ALI/cALI, and sES levels decreased following treatment of their pneumonia. Univariate and multivariate logistic regression analyses showed that sES was the only significant factor for identifying complicating ALI/cALI, independently of C-reactive protein (CRP and lactate dehydrogenase (LDH. By receiver operating characteristic (ROC curve analysis, the cut-off value for sES was 40.1 ng/mL, with a sensitivity of 0.8 and a specificity of 0.8.Conclusion: sES may be a useful biomarker for discriminating complicating ALI/cALI in patients with severe pneumonia.

  7. Inhibition of Phosphodiesterase-4 during Pneumococcal Pneumonia Reduces Inflammation and Lung Injury in Mice.

    Science.gov (United States)

    Tavares, Luciana P; Garcia, Cristiana C; Vago, Juliana P; Queiroz-Junior, Celso M; Galvão, Izabela; David, Bruna A; Rachid, Milene A; Silva, Patrícia M R; Russo, Remo C; Teixeira, Mauro M; Sousa, Lirlândia P

    2016-07-01

    Pneumococcal pneumonia is a leading cause of mortality worldwide. The inflammatory response to bacteria is necessary to control infection, but it may also contribute to tissue damage. Phosphodiesterase-4 inhibitors, such as rolipram (ROL), effectively reduce inflammation. Here, we examined the impact of ROL in a pneumococcal pneumonia murine model. Mice were infected intranasally with 10(5)-10(6) CFU of Streptococcus pneumoniae, treated with ROL in a prophylactic or therapeutic schedule in combination, or not, with the antibiotic ceftriaxone. Inflammation and bacteria counts were assessed, and ex vivo phagocytosis assays were performed. ROL treatment during S. pneumoniae infection decreased neutrophil recruitment into lungs and airways and reduced lung injury. Prophylactic ROL treatment also decreased cytokine levels in the airways. Although modulation of inflammation by ROL ameliorated pneumonia, bacteria burden was not reduced. On the other hand, antibiotic therapy reduced bacteria without reducing neutrophil infiltration, cytokine level, or lung injury. Combined ROL and ceftriaxone treatment decreased lethality rates and was more efficient in reducing inflammation, by increasing proresolving protein annexin A1 (AnxA1) expression, and bacterial burden by enhancing phagocytosis. Lack of AnxA1 increased inflammation and lethality induced by pneumococcal infection. These data show that immunomodulatory effects of phosphodiesterase-4 inhibitors are useful during severe pneumococcal pneumonia and suggest their potential benefit as adjunctive therapy during infectious diseases.