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Sample records for sccmec types iv

  1. A new multiplex PCR for easy screening of methicillin-resistant Staphylococcus aureus SCCmec types I-V

    DEFF Research Database (Denmark)

    Boye, Kit; Bartels, Mette Damkjær; Andersen, Ina S

    2007-01-01

    A multiplex PCR with four primer-pairs was designed to identify the five main known SCCmec types. A clear and easily discriminated band pattern was obtained for all five types. The SCCmec type was identified for 98% of 312 clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA......). SCCmec type IV was by far the most common SCCmec type among both hospital- and community-acquired MRSA isolates in Denmark....

  2. Characteristics of SCCmec IV and V Methicillin-Resistant Staphylococcus aureus (MRSA) in Israel.

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    Shitrit, Pnina; Openhaim, Michal; Reisfeld, Sharon; Paitan, Yossi; Regev-Yochay, Gili; Carmeli, Yehuda; Chowers, Michal

    2015-08-01

    Isolation of methicillin-resistant Staphylococcus aureus (MRSA) in healthy individuals is not common in Israel. In our hospital, about 30% of MRSA isolates were SCCmec types IV and V. To identify the demographic and clinical characteristics of patients carrying MRSA SCCmec type IV or V, and to compare them with each other and with those of patients with SCCmec types I-III. We conducted a case-control study that included 501 patients from whom MRSA was isolated: 254 with SCCmec type I, II, or III, and 243 isolates from SCCmec types IV or V. MRSA was isolated from surveillance cultures in 75% of patients and from a clinical site in 25%. The majority of our study population was elderly, from nursing homes, and with extensive exposure to health care. First, we compared characteristics of patients identified through screening. Statistically significant predictors of SCCmec V vs. IV were Arab ethnicity (OR 7.44, 95% CI 1.5-37.9) and hospitalization in the year prior to study inclusion (OR 5.7, 95% CI 1.9-16.9). No differences were found between patients with SCCmec types I-III and patients with SCCmec type IV or V. Analysis of the subset of patients who had clinical cultures yielded similar results. SCCmec types IV and V were common in the hospital setting although rare in the community. It seems that in Israel, SCCmec IV and V are predominantly health care-associated MRSA.

  3. Predominance of Three Closely Related Methicillin-Resistant Staphylococcus aureus Clones Carrying a Unique ccrC-Positive SCCmec type III and the Emergence of spa t304 and t690 SCCmec type IV pvl+MRSA Isolates in Kinta Valley, Malaysia.

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    Ho, Wai-Yew; Choo, Quok-Cheong; Chew, Choy-Hoong

    2017-03-01

    We investigated the epidemiology and clonality of 175 nonrepetitive methicillin-resistant Staphylococcus aureus (MRSA) isolates from clinical specimens collected between 2011 and 2012 in Kinta Valley in Malaysia. Molecular tools such as polymerase chain reaction, pulsed-field gel electrophoresis, and staphylococcal protein A (spa) typing were used. Our study revealed the predominance of three closely related ermA + SCCmec type III pulsotypes belonging to spa type t037 (Brazilian-Hungarian clone), which were deficient in the locus F, but positive for the ccrC gene in majority (65.7%) of the MRSA infections in this region. The first evidence of SCCmec type II MRSA in the country, belonging to spa type t2460, was also noted. Although the carriage of pvl gene was uncommon (8.6%) and mostly confined to either SCCmec type IV or SCCmec type V isolates, most of these isolates belonged to spa types t345 or t657, which are associated with the Bengal-Bay CA-MRSA clone. Interestingly, spa t304 and t690 SCCmec type IV pvl + were also detected among the MRSA isolates. Data from this study show the rise of uncommon clones among MRSA isolates in Malaysia.

  4. Community-acquired methicillin-resistant Staphylococcus aureus carrying SCCmec type IV and V isolated from healthy children attending public daycares in northeastern Brazil.

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    Carvalho, Suzi P de; Almeida, Jéssica B de; Andrade, Yasmin M F S; Silva, Lucas S C da; Oliveira, Arianne C de; Nascimento, Flávia S; Campos, Guilherme B; Oliveira, Márcio V; Timenetsky, Jorge; Marques, Lucas M

    Nasal colonization with methicillin-resistant Staphylococcus aureus (MRSA) have increasingly been reported in healthy communities. This study aimed to assess the rate of S. aureus in general and MRSA in particular from nasal secretion of children in daycare centers in Vitória da Conquista, Brazil. The isolates were identified based on morphology, biochemical tests and by PCR. Detection of virulence genes, biofilm production, and susceptibility test by disk diffusion agar were performed. MRSA isolates were characterized by spa, SCCmec, and multilocus sequence typing (MLST). S. aureus were recovered from 70 (47.3%) of 148 children. Among the 11 MRSA strains (15.7%), two SCCmec types (IV and V) were detected. MLST identified four STs related to three clonal complexes (CC): 5, 45, and 398. Four spa types were found circulating in this setting. Resistance of S. aureus isolates to ampicillin, erythromycin, ciprofloxacin, clindamycin, and tetracycline was 80%, 32.8%, 7.1%, 7.1% and 4.3%, respectively. One isolate presented intermediate resistance to vancomycin detected by Etest methodology. All strains were biofilm producers. The virulence genes seb, sec, spa, and pvl were detected in some isolates. This study revealed a high rate of children carrying MRSA among healthy attendees in daycare centers in Vitória da Conquista, Brazil. Copyright © 2017 Sociedade Brasileira de Infectologia. Published by Elsevier Editora Ltda. All rights reserved.

  5. Community-acquired methicillin-resistant Staphylococcus aureus carrying SCCmec type IV and V isolated from healthy children attending public daycares in northeastern Brazil

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    Suzi P. de Carvalho

    2017-07-01

    Full Text Available Nasal colonization with methicillin-resistant Staphylococcus aureus (MRSA have increasingly been reported in healthy communities. This study aimed to assess the rate of S. aureus in general and MRSA in particular from nasal secretion of children in daycare centers in Vitória da Conquista, Brazil. The isolates were identified based on morphology, biochemical tests and by PCR. Detection of virulence genes, biofilm production, and susceptibility test by disk diffusion agar were performed. MRSA isolates were characterized by spa, SCCmec, and multilocus sequence typing (MLST. S. aureus were recovered from 70 (47.3% of 148 children. Among the 11 MRSA strains (15.7%, two SCCmec types (IV and V were detected. MLST identified four STs related to three clonal complexes (CC: 5, 45, and 398. Four spa types were found circulating in this setting. Resistance of S. aureus isolates to ampicillin, erythromycin, ciprofloxacin, clindamycin, and tetracycline was 80%, 32.8%, 7.1%, 7.1% and 4.3%, respectively. One isolate presented intermediate resistance to vancomycin detected by Etest methodology. All strains were biofilm producers. The virulence genes seb, sec, spa, and pvl were detected in some isolates. This study revealed a high rate of children carrying MRSA among healthy attendees in daycare centers in Vitória da Conquista, Brazil.

  6. Associations between dru Types and SCCmec Cassettes

    DEFF Research Database (Denmark)

    Bartels, Mette D; Boye, Kit; Oliveira, Duarte C

    2013-01-01

    isolates with dru type variants indicating that dru typing is not useful as a first line epidemiological typing tool. However, MRSA isolates cultured from a single patient over a three year period exhibited a single dru type. The finding of dt10a in most SCCmec types suggests that dru and mecA originate......Molecular typing is an important tool in the investigation of methicillin resistant Staphylococcus aureus (MRSA) outbreaks and in following the evolution of MRSA. The staphylococcal cassette chromosome mec (SCCmec) contains a hypervariable region with a variable number of 40 bp repeats named direct...... repeat units (dru). The dru region has been suggested as a supplementary typing method for MRSA and an international nomenclature exists. The purpose of this study was to investigate the diversity and variability of the dru region in a diverse collection of MRSA. We studied 302 MRSA isolates harbouring...

  7. [Differentiation of spa types and staphylococcal cassette chromosome mec (SCCmec) in clinical methicillin-resistant Staphylococcus aureus isolated in medical sites of Gdańsk region].

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    Kasprzyk, Joanna; Piechowicz, Lidia; Wiśniewska, Katarzyna; Dziewit, Łukasz; Bronk, Marek; Świeć, Krystyna

    2015-01-01

    Methicillin-resistant Staphylococcus aureus bacteria are one of the key etiological factors of hospital-acquired and community-acquired infections. MRSA strains have an ability of causing a broad spectrum infections: from a relatively mild skin infections to severe life-threatening systemic infections. They are characterized by multi-drug resistance, virulence of a number of factors, may clonally spread within the hospitals and between hospitals. The study embraced a number of 75 isolates of MRSA isolated from patients of 7 medical sites of the Gdansk region within the period of six months (June to December 2013). Strains have derived from various clinical materials, both of hospitalized patients (n=59) and outpatient (n=16). The isolates were tested for the susceptibility to antimicrobial agents accordance with the guidelines EUCAST. To estimate of the variability of occurrence of S. aureus clones used were standard spa gene, consisting in the amplified polymorphic region of the X gene encoding the protein A gene (spa). After receiving the results, a spa types were identified using international database Ridom Spa Server (www.spaserver.ridom.de). To determine the polymorphism cassette carrying the inecA gene from MRSA strains, used typing five major chromosomal cassette SCCmec (I-V) by multiplex PCR. MRSA population genetic analysis carried out on the basis of typing SCCmec cassettes and spa gene has showed a predominance of strains with SCCmec type II casette (46.7%) and SCCmec IV casette (38.7%). Less frequently detected were strains containing SCCmec I cassette (12.0%) and SCCmec III cassette (2.6%). Spa typing revealed the presence of 13 gene types in MRSA. The most frequently observed spa types were: t151 (24.0%), t003 (16.0%) in strains of the SCCmec II cassette and t437 (16.0%) and t008 (14.8%) in the isolates with SCCmec cassette IV, whereas staphylococcus with the type of spa t011 (12.0%) had SCCmec cassette I. In our population most frequent strains

  8. The dissemination of ST80-SCCmec-IV community-associated methicillin resistant Staphylococcus aureus clone in Kuwait hospitals

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    Sarkhoo Eiman

    2010-11-01

    Full Text Available Abstract Background Community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA is a global healthcare problem. The purpose of this study was to characterize CA-MRSA clones and their distribution in Kuwait hospitals. Methods In total, 135 CA-MRSA isolates, carrying the SCCmec IV or V genetic elements, isolated in eight hospitals were characterized using antibiogram, pulsed-field gel electrophoresis, multilocus sequence typing, and carriage of genes for Panton-Valentine Leukocidin (PVL, capsular polysaccharides types (cap 5 and 8, accessory genes regulators (agr, Staphylococcal enterotoxins (SE and toxic shock syndrome toxin 1 (tst. Results They were susceptible to vancomycin, teicoplanin and linezolid but resistant to kanamycin (62%, fusidic acid (42.2%, tetracycline (39.3%, erythromycin and clindamycin (21.5%, gentamicin (5.9%, streptomycin (6.7%, trimethoprim (5.9%, mupirocin (6.6% and cadmium acetate (82.2%. They consisted of 10 pulsotypes with the majority belonging to PFGE type I (51.1%, type II (22.2%, type IV (13.3% and type III (3.7%. They belonged to 10 sequence types (ST comprising ST80 (51.1%, ST30 (22.2%, ST5 (14.1%, ST1 (4.45, ST6 (3.7%, ST88 (1.5%, ST834 (1.5%, ST8 (0.7%, ST46 (0.7% and ST950 (0.7%. Genes for PVL, cap 8, cap 5 and agr III, agr I and agr II were detected in 61.5%, 77.3%, 20.7% and 62.2%, 17% and 8.1% of the isolates respectively. Nine (6.7% isolates contained tst while 103 isolates were positive for SE genes with sei (63.0%, seg (41.5% and sed (29.6% as the common SE genes. Conclusions ST80-SCCmecIV was the most common CA-MRSA clone in Kuwait hospitals presenting new challenges for infection control.

  9. Complete Circular Genome Sequence of Successful ST8/SCCmecIV Community-Associated Methicillin-Resistant Staphylococcus aureus (OC8) in Russia: One-Megabase Genomic Inversion, IS256's Spread, and Evolution of Russia ST8-IV.

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    Wan, Tsai-Wen; Khokhlova, Olga E; Iwao, Yasuhisa; Higuchi, Wataru; Hung, Wei-Chun; Reva, Ivan V; Singur, Olga A; Gostev, Vladimir V; Sidorenko, Sergey V; Peryanova, Olga V; Salmina, Alla B; Reva, Galina V; Teng, Lee-Jene; Yamamoto, Tatsuo

    2016-01-01

    ST8/SCCmecIV community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) has been a common threat, with large USA300 epidemics in the United States. The global geographical structure of ST8/SCCmecIV has not yet been fully elucidated. We herein determined the complete circular genome sequence of ST8/SCCmecIVc strain OC8 from Siberian Russia. We found that 36.0% of the genome was inverted relative to USA300. Two IS256, oppositely oriented, at IS256-enriched hot spots were implicated with the one-megabase genomic inversion (MbIN) and vSaβ split. The behavior of IS256 was flexible: its insertion site (att) sequences on the genome and junction sequences of extrachromosomal circular DNA were all divergent, albeit with fixed sizes. A similar multi-IS256 system was detected, even in prevalent ST239 healthcare-associated MRSA in Russia, suggesting IS256's strong transmission potential and advantage in evolution. Regarding epidemiology, all ST8/SCCmecIVc strains from European, Siberian, and Far Eastern Russia, examined had MbIN, and geographical expansion accompanied divergent spa types and resistance to fluoroquinolones, chloramphenicol, and often rifampicin. Russia ST8/SCCmecIVc has been associated with life-threatening infections such as pneumonia and sepsis in both community and hospital settings. Regarding virulence, the OC8 genome carried a series of toxin and immune evasion genes, a truncated giant surface protein gene, and IS256 insertion adjacent to a pan-regulatory gene. These results suggest that unique single ST8/spa1(t008)/SCCmecIVc CA-MRSA (clade, Russia ST8-IVc) emerged in Russia, and this was followed by large geographical expansion, with MbIN as an epidemiological marker, and fluoroquinolone resistance, multiple virulence factors, and possibly a multi-IS256 system as selective advantages.

  10. Characterization of SCCmec and spa types of methicillin-resistant Staphylococcus aureus isolates from health-care and community-acquired infections in Kerman, Iran.

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    Fasihi, Yaser; Kiaei, Somayeh; Kalantar-Neyestanaki, Davood

    2017-12-01

    Spread of methicillin-resistant Staphylococcus aureus (MRSA) isolates is a worldwide problem. Molecular typing is a useful tool to understand MRSA epidemiology. Herein, we determined vancomycin-resistant, SCCmec and spa types among MRSA isolates recovered from healthcare and community-acquired infections in Kerman, Iran. A total of 170 S. aureus isolates were collected from different patients who were admitted to affiliated hospitals of Kerman University of Medical science. MRSA and vancomycin-resistant S. aureus (VRSA) isolates were detected by phenotypic methods. Polymerase chain reaction (PCR) technique was used for detection of mecA, vanA and vanB genes. Staphylococcal cassette chromosomemec (SCCmec) and spa typing were used for molecular typing of among MRSA isolates. Overall, 53% of isolates were considered as MRSA. Two MRSA isolates were resistant to vancomycin and vanA was detected in only one of VRSA isolates. SCCmec type III belonged to spa types t030 and t459 which they were the dominant spa types among community-associated MRSA (CA-MRSA) and healthcare-acquired MRSA (HA-MRSA) isolates. Our findings showed that the SCCmec type I and III spread from hospital settings to community, although the SCCmec type IV spread from community to healthcare systems. We have also reported VRSA isolates from hospitalized patients, therefore, appropriate policies should be enforced in order to prevent the spread of antibiotic resistance isolates in hospitals settings. Copyright © 2017 Ministry of Health, Saudi Arabia. Published by Elsevier Ltd. All rights reserved.

  11. Predominant Dissemination of PVL-Negative CC89 MRSA with SCCmec Type II in Children with Impetigo in Japan

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    H. Kikuta

    2011-01-01

    Full Text Available Background. The ratio of CA-MRSA in children with impetigo has been increasing in Japan. Methods. Antimicrobial susceptibilities of 136 S. aureus isolates from children with impetigo were studied. Furthermore, molecular epidemiological analysis and virulence gene analysis were performed. Results. Of the 136 S. aureus isolates, 122 (89.7% were MSSA and 14 (10.3% were MRSA. Of the 14 MRSA strains, 11 belonged to CC89 (ST89, ST91, and ST2117 and carried diverse types of SCCmec: type II (IIb: 3 strains; unknown subtype: 4 strains, type IVa (2 strains, and unknown type (2 strains. The remaining three strains exhibited CC8 (ST-8-SCCmec type VIa, CC121 (ST121-SCCmec type V, and CC5 (ST5-nontypeable SCCmec element, respectively. None were lukS-PV-lukF-PV gene positive. Gentamicin- and clarithromycin-resistant strains were frequently found in both MRSA and MSSA. Conclusions. PVL-negative CC89-SCCmec type II strains are the most predominant strains among the CA-MRSA strains circulating in the community in Japan.

  12. Complete Circular Genome Sequence of Successful ST8/SCCmecIV Community-Associated Methicillin-Resistant Staphylococcus aureus (OC8) in Russia: One-Megabase Genomic Inversion, IS256’s Spread, and Evolution of Russia ST8-IV

    Science.gov (United States)

    Wan, Tsai-Wen; Higuchi, Wataru; Hung, Wei-Chun; Reva, Ivan V.; Singur, Olga A.; Gostev, Vladimir V.; Sidorenko, Sergey V.; Peryanova, Olga V.; Salmina, Alla B.; Reva, Galina V.; Teng, Lee-Jene; Yamamoto, Tatsuo

    2016-01-01

    ST8/SCCmecIV community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) has been a common threat, with large USA300 epidemics in the United States. The global geographical structure of ST8/SCCmecIV has not yet been fully elucidated. We herein determined the complete circular genome sequence of ST8/SCCmecIVc strain OC8 from Siberian Russia. We found that 36.0% of the genome was inverted relative to USA300. Two IS256, oppositely oriented, at IS256-enriched hot spots were implicated with the one-megabase genomic inversion (MbIN) and vSaβ split. The behavior of IS256 was flexible: its insertion site (att) sequences on the genome and junction sequences of extrachromosomal circular DNA were all divergent, albeit with fixed sizes. A similar multi-IS256 system was detected, even in prevalent ST239 healthcare-associated MRSA in Russia, suggesting IS256’s strong transmission potential and advantage in evolution. Regarding epidemiology, all ST8/SCCmecIVc strains from European, Siberian, and Far Eastern Russia, examined had MbIN, and geographical expansion accompanied divergent spa types and resistance to fluoroquinolones, chloramphenicol, and often rifampicin. Russia ST8/SCCmecIVc has been associated with life-threatening infections such as pneumonia and sepsis in both community and hospital settings. Regarding virulence, the OC8 genome carried a series of toxin and immune evasion genes, a truncated giant surface protein gene, and IS256 insertion adjacent to a pan-regulatory gene. These results suggest that unique single ST8/spa1(t008)/SCCmecIVc CA-MRSA (clade, Russia ST8-IVc) emerged in Russia, and this was followed by large geographical expansion, with MbIN as an epidemiological marker, and fluoroquinolone resistance, multiple virulence factors, and possibly a multi-IS256 system as selective advantages. PMID:27741255

  13. Molecular characteristics of methicillin-resistant Staphylococcus aureus (MRSA) strains isolated from patients with bacteremia based on MLST, SCCmec, spa, and agr locus types analysis.

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    Goudarzi, Mehdi; Seyedjavadi, Sima Sadat; Nasiri, Mohammad Javad; Goudarzi, Hossein; Sajadi Nia, Raheleh; Dabiri, Hossein

    2017-03-01

    The widespread emergence of methicillin resistant Staphylococcus aureus, as a common cause of nosocomial infections, is becoming a serious concern in global public health. The objective of the present study was to investigate antimicrobial susceptibility pattern, frequency of virulence genes and molecular characteristics of methicillin-resistant Staphylococcus aureus strains isolated from patients with bacteremia. A total of 128 methicillin-resistant Staphylococcus aureus isolates were collected during February 2015 to January 2016. In vitro antimicrobial susceptibility of the isolates was assessed using the disk diffusion method. Conventional PCR was performed for the detection of adhesion (can, bbp, ebp, fnbB, fnbA, clfB, clfA) and toxin (etb, eta, pvl, tst) encoding genes, determining the agr type, SCCmec, MLST and spa typing of the isolates. All the methicillin-resistant Staphylococcus aureus isolates were found to be sensitive to linezolid, teicoplanin, and vancomycin. Resistance to the tested antibiotics varied from 97.7% for penicillin to 24.2% for mupirocin. The rate of multi drug resistance (MDR) in the present study was 97.7%. The most commonly detected toxin and adhesion genes were tst (58.6%), and clfB (100%), respectively. The majority of SCCmec III isolates were found in agr group I while SCCmec IV and II isolates were distributed among agr group III. Multilocus Sequence Typing (MLST) of the MRSA isolates showed five different sequence types: ST239 (43%), ST22 (39.8%), ST585 (10.9%), ST45 (3.9%) and ST240 (2.3%). All of the pvl positive strains belonged to ST22-SCCmec IV/t790 clone and were MDR. Among different 7 spa types, the most common were t790 (27.3%), t037 (21.9%), and t030 (14.1%). spa types t016, t924 and spa type t383 were reported for the first time from Asia and Iran, respectively. It was shown that spa types circulating in the studied hospitals varied which support the need to perform future surveillance studies in order to understand

  14. DNA microarray profiling of a diverse collection of nosocomial methicillin-resistant staphylococcus aureus isolates assigns the majority to the correct sequence type and staphylococcal cassette chromosome mec (SCCmec) type and results in the subsequent identification and characterization of novel SCCmec-SCCM1 composite islands.

    LENUS (Irish Health Repository)

    Shore, Anna C

    2012-10-01

    One hundred seventy-five isolates representative of methicillin-resistant Staphylococcus aureus (MRSA) clones that predominated in Irish hospitals between 1971 and 2004 and that previously underwent multilocus sequence typing (MLST) and staphylococcal cassette chromosome mec (SCCmec) typing were characterized by spa typing (175 isolates) and DNA microarray profiling (107 isolates). The isolates belonged to 26 sequence type (ST)-SCCmec types and subtypes and 35 spa types. The array assigned all isolates to the correct MLST clonal complex (CC), and 94% (100\\/107) were assigned an ST, with 98% (98\\/100) correlating with MLST. The array assigned all isolates to the correct SCCmec type, but subtyping of only some SCCmec elements was possible. Additional SCCmec\\/SCC genes or DNA sequence variation not detected by SCCmec typing was detected by array profiling, including the SCC-fusidic acid resistance determinant Q6GD50\\/fusC. Novel SCCmec\\/SCC composite islands (CIs) were detected among CC8 isolates and comprised SCCmec IIA-IIE, IVE, IVF, or IVg and a ccrAB4-SCC element with 99% DNA sequence identity to SCC(M1) from ST8\\/t024-MRSA, SCCmec VIII, and SCC-CI in Staphylococcus epidermidis. The array showed that the majority of isolates harbored one or more superantigen (94%; 100\\/107) and immune evasion cluster (91%; 97\\/107) genes. Apart from fusidic acid and trimethoprim resistance, the correlation between isolate antimicrobial resistance phenotype and the presence of specific resistance genes was ≥97%. Array profiling allowed high-throughput, accurate assignment of MRSA to CCs\\/STs and SCCmec types and provided further evidence of the diversity of SCCmec\\/SCC. In most cases, array profiling can accurately predict the resistance phenotype of an isolate.

  15. Update to the multiplex PCR strategy for assignment of mec element types in Staphylococcus aureus.

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    Milheiriço, Catarina; Oliveira, Duarte C; de Lencastre, Hermínia

    2007-09-01

    Staphylococcal cassette chromosome mec (SCCmec) typing is important for the identification and definition of methicillin-resistant Staphylococcus aureus clones, and for routine purposes, multiplex PCR assays are the most adequate for SCCmec typing. Here, we describe an update to the multiplex PCR strategy for SCCmec typing that we described in 2002 so that SCCmec types IV and V may be properly identified.

  16. First report on methicillin-resistant Staphylococcus aureus of Spa type T037, Sequence type 239, SCCmec type III/IIIA in Malaysia

    NARCIS (Netherlands)

    V. Neela (Vasanthakumari); H. Ghasemzadeh Moghaddam (Hamed); A.F. van Belkum (Alex); D. Horst-Kreft (Deborah); N.S. Mariana (Nor Shamsudin); E. Ghaznavi Rad (Ehsanollah)

    2010-01-01

    textabstractMethicillin-resistant Staphylococcus aureus (MRSA) from Malaysia were shown to possess staphylococcal cassette chromosome mec (SCCmec)-III and IIIA. Spa sequencing and multi-locus sequence typing (MLST) documented t037 and ST 239 (CC8) for 83.3% of the isolates. This confirms

  17. Whole-genome comparison of meticillin-resistant Staphylococcus aureus CC22 SCCmecIV from people and their in-contact pets.

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    Loeffler, Anette; McCarthy, Alex; Lloyd, David H; Musilová, Eva; Pfeiffer, Dirk U; Lindsay, Jodi A

    2013-10-01

    Meticillin-resistant Staphylococcus aureus (MRSA) infections remain important medical and veterinary challenges. The MRSA isolated from dogs and cats typically belong to dominant hospital-associated clones, in the UK mostly EMRSA-15 (CC22 SCCmecIV), suggesting original human-to-animal transmission. Nevertheless, little is known about host-specific genetic variation within the same S. aureus lineage. To identify host-specific variation amongst MRSA CC22 SCCmecIV by comparing isolates from pets with those from in-contact humans using whole-genome microarray. Six pairs of MRSA CC22 SCCmecIV from human carriers (owners and veterinary staff) and their respective infected in-contact pets were compared using a 62-strain whole-genome S. aureus microarray (SAM-62). The presence of putative host-specific genes was subsequently determined in a larger number of human (n = 47) and pet isolates (n = 93) by PCR screening. Variation in mobile genetic elements (MGEs) occurred frequently and appeared largely independent of host and in-contact pair. A plasmid (SAP078A) encoding heavy-metal resistance genes (arsR, arsA, cadA, cadC, mco and copB) was found in three of six human and none of six animal isolates. However, only two of four resistance genes were associated with human hosts (P = 0.015 for arsA and cadA). The variation found amongst MGEs highlights that genetic adaptation in MRSA continues. However, host-specific MGEs were not detected, which supports the hypothesis that pets may not be natural hosts of MRSA CC22 and emphasizes that rigorous hygiene measures are critical to prevent contamination and infection of dogs and cats. The host specificity of individual heavy-metal resistance genes warrants further investigation into different selection pressures in humans and animals. © 2013 ESVD and ACVD.

  18. SCCmec types and pvl gene in methicillin-resistant Staphylococcus aureus strains from children hospitalized in a tertiary care hospital in Mexico.

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    Borbón-Esquer, Eunice Mireya; Villaseñor-Sierra, Alberto; Martínez-López, Erika; Jáuregui-Lomeli, Juan José; Villaseñor-Martínez, Rosa; Ruiz-Briseño, Mariana Del Rocío

    2014-07-01

    The aim of this study was to determine the prevalence, SCCmec types, presence of the Panton-Valentine leukocidin (PVL) gene, and susceptibility to antibiotics of methicillin-resistant Staphylococcus aureus (MRSA) strains isolated from hospitalized children. From August 2009 to September 2011, 291 S. aureus strains were isolated from normally sterile body sites, of which 190 (65%) were MRSA. One hundred and two of the MRSA strains were genetically evaluated. SCCmec genotypes were identified by M-PCR and the PVL gene (pvl) by end-point PCR. Resistance to erythromycin, rifampicin, clindamycin, and trimethoprim-sulfamethoxazole (SXT) was assessed by Kirby-Bauer disk diffusion method in accordance with the Clinical and Laboratory Standards Institute guidelines of 2012. Of the 102 strains evaluated, 97 (95%) were SCCmec type II, 5 (5%) were SCCmec type IVa, and all (100%) were pvl-negative. Resistance to erythromycin, clindamycin, rifampicin, and SXT was 97%, 95%, 0%, and 0%, respectively. The prevalence of hospital-acquired MRSA was high. SCCmec type II was predominant and the pvl gene appeared not to play any role in the virulence of the MRSA strains from hospitalized children.

  19. Comparison of spa types, SCCmec types and antimicrobial resistance profiles of MRSA isolated from turkeys at farm, slaughter and from retail meat indicates transmission along the production chain.

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    Birgit Vossenkuhl

    Full Text Available The prevalence of MRSA in the turkey meat production chain in Germany was estimated within the national monitoring for zoonotic agents in 2010. In total 22/112 (19.6% dust samples from turkey farms, 235/359 (65.5% swabs from turkey carcasses after slaughter and 147/460 (32.0% turkey meat samples at retail were tested positive for MRSA. The specific distributions of spa types, SCCmec types and antimicrobial resistance profiles of MRSA isolated from these three different origins were compared using chi square statistics and the proportional similarity index (Czekanowski index. No significant differences between spa types, SCCmec types and antimicrobial resistance profiles of MRSA from different steps of the German turkey meat production chain were observed using Chi-Square test statistics. The Czekanowski index which can obtain values between 0 (no similarity and 1 (perfect agreement was consistently high (0.79-0.86 for the distribution of spa types and SCCmec types between the different processing stages indicating high degrees of similarity. The comparison of antimicrobial resistance profiles between the different process steps revealed the lowest Czekanowski index values (0.42-0.56. However, the Czekanowski index values were substantially higher than the index when isolates from the turkey meat production chain were compared to isolates from wild boar meat (0.13-0.19, an example of a separated population of MRSA used as control group. This result indicates that the proposed statistical method is valid to detect existing differences in the distribution of the tested characteristics of MRSA. The degree of similarity in the distribution of spa types, SCCmec types and antimicrobial resistance profiles between MRSA isolates from different process stages of turkey meat production may reflect MRSA transmission along the chain.

  20. High frequency of methicillin-resistant Staphylococcus aureus (MRSA) with SCCmec type III and spa type t030 in Karaj's teaching hospitals, Iran.

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    Bayat, Bahareh; Zade, Masoumeh Hallaj; Mansouri, Samaneh; Kalantar, Enayat; Kabir, Kourosh; Zahmatkesh, Ehsan; Sepehr, Mohammad Noori; Naseri, Mohammmad Hassan; Darban-Sarokhalil, Davood

    2017-09-01

    Methicillin-resistant Staphylococcus aureus (MRSA) has been one of the most important antibiotic-resistant pathogen in many parts of the world over the past decades. This cross-sectional study was conducted to investigate MRSA isolated between July 2013 and July 2014 in Karaj, Iran. All tested isolates were collected in teaching hospitals from personnel, patients, and surfaces and each MRSA was analyzed by SCCmec and spa typing. Antibiotic susceptibility testing was accomplished by disk diffusion method. Out of 49 MRSA isolates from the Karaj's teaching hospitals, 82%, 10%, and 6% of the isolates were SCCmec types III, II, and I, respectively. The main spa type in this study was spa t030 with frequency as high as 75.5% from intensive care unit (ICU) of the hospitals and high rate of resistance to rifampicin (53%) was found in MRSA isolates. In conclusion, high frequency of spa t030 with SCCmec type III and MRSA phenotype illustrated circulating of one of the antibiotic-resistant strains in ICU of Karaj's teaching hospitals and emphasizes the need for ongoing molecular surveillance, antibiotic susceptibility monitoring, and infection control.

  1. Comparison of spa Types, SCCmec Types and Antimicrobial Resistance Profiles of MRSA Isolated from Turkeys at Farm, Slaughter and from Retail Meat Indicates Transmission along the Production Chain

    OpenAIRE

    Vossenkuhl, Birgit; Brandt, Jörgen; Fetsch, Alexandra; Käsbohrer, Annemarie; Kraushaar, Britta; Alt, Katja; Tenhagen, Bernd-Alois

    2014-01-01

    The prevalence of MRSA in the turkey meat production chain in Germany was estimated within the national monitoring for zoonotic agents in 2010. In total 22/112 (19.6%) dust samples from turkey farms, 235/359 (65.5%) swabs from turkey carcasses after slaughter and 147/460 (32.0%) turkey meat samples at retail were tested positive for MRSA. The specific distributions of spa types, SCCmec types and antimicrobial resistance profiles of MRSA isolated from these three different origins were compare...

  2. Emergence of sequence type 779 methicillin-resistant Staphylococcus aureus harboring a novel pseudo staphylococcal cassette chromosome mec (SCCmec)-SCC-SCCCRISPR composite element in Irish hospitals.

    LENUS (Irish Health Repository)

    Kinnevey, Peter M

    2013-01-01

    Methicillin-resistant Staphylococcus aureus (MRSA) has been a major cause of nosocomial infection in Irish hospitals for 4 decades, and replacement of predominant MRSA clones has occurred several times. An MRSA isolate recovered in 2006 as part of a larger study of sporadic MRSA exhibited a rare spa (t878) and multilocus sequence (ST779) type and was nontypeable by PCR- and DNA microarray-based staphylococcal cassette chromosome mec (SCCmec) element typing. Whole-genome sequencing revealed the presence of a novel 51-kb composite island (CI) element with three distinct domains, each flanked by direct repeat and inverted repeat sequences, including (i) a pseudo SCCmec element (16.3 kb) carrying mecA with a novel mec class region, a fusidic acid resistance gene (fusC), and two copper resistance genes (copB and copC) but lacking ccr genes; (ii) an SCC element (17.5 kb) carrying a novel ccrAB4 allele; and (iii) an SCC element (17.4 kb) carrying a novel ccrC allele and a clustered regularly interspaced short palindromic repeat (CRISPR) region. The novel CI was subsequently identified by PCR in an additional 13 t878\\/ST779 MRSA isolates, six from bloodstream infections, recovered between 2006 and 2011 in 11 hospitals. Analysis of open reading frames (ORFs) carried by the CI showed amino acid sequence similarity of 44 to 100% to ORFs from S. aureus and coagulase-negative staphylococci (CoNS). These findings provide further evidence of genetic transfer between S. aureus and CoNS and show how this contributes to the emergence of novel SCCmec elements and MRSA strains. Ongoing surveillance of this MRSA strain is warranted and will require updating of currently used SCCmec typing methods.

  3. SCCmec Type IX Element in Methicillin Resistant Staphylococcusaureusspa Type t337 (CC9) Isolated from Pigs and Pork in Thailand

    DEFF Research Database (Denmark)

    Vestergaard, Martin; Cavaco, Lina M; Sirichote, Pantip

    2012-01-01

    blaZ, aac-aphD, vga(A), tetM, and a tet efflux marker, in all strains and additionally ermB and aadD, cat and fex(A) in the pork isolates. None of the isolates were found PVL-positive, but enterotoxins were identified in all isolates. To our knowledge, only a few descriptions of MRSA in livestock...... and food products in Thailand have been observed but this is the first observation of MRSA CC9 associated with SCCmec IX in pork. This study indicates a likely widespread distribution of MRSA in pig and pork in Thailand and further investigation on the prevalence and importance of livestock associated MRSA...

  4. Genetic diversity of methicillin resistant Staphylococcus aureus strains isolated from burn patients in Iran: ST239-SCCmec III/t037 emerges as the major clone.

    Science.gov (United States)

    Goudarzi, Mehdi; Bahramian, Mahnaz; Satarzadeh Tabrizi, Mahboobeh; Udo, Edet E; Figueiredo, Agnes Marie Sá; Fazeli, Maryam; Goudarzi, Hossein

    2017-04-01

    Methicillin-resistant Staphylococcus aureus (MRSA) as a major cause of infection in health care, hospital and community settings is a global health concern. The purpose of this study was to determine the antibiotic susceptibility pattern and distribution of circulating molecular types of MRSA in a burn hospital in Tehran, the capital of Iran. During a 10-month study period, 106 Staphylococcus aureus isolates were assessed. Isolates were subjected to susceptibility testing using the disk diffusion method and Polymerase Chain Reaction (PCR) for detection of mecA, fem and nuc genes. The presence of PVL and tst encoding genes were determined by PCR method. All the MRSA isolates were genotyped by multilocus sequence typing (MLST), spa typing, SCCmec typing and agr typing. The presence of mecA gene was confirmed in all the Staphylococcus aureus isolates. Antimicrobial susceptibility testing revealed a high resistance rate (90.6%) to ampicillin, tetracycline, and erythromycin. The rates of resistance to remaining antibiotics tested varied between 18.9% and 84.9%. The high- level of resistance to mupirocin was confirmed in 19.8% of MRSA strains isolated from burn patients. Multi-drug resistance was observed in 90.6% of isolates. Sixteen of the 106 MRSA isolates (15.1%) harbored PVL-encoding genes. The majority of our MRSA strains carried SCCmec III (71.7%). ST239-SCCmec III/t037 (34%) was the most common genotype followed by ST239-SCCmec III/t030 (24.5%), ST15-SCCmec IV/t084 (15.1%), ST22-SCCmec IV/t790 (13.2%), and ST239-SCCmec III/t631 (13.2%). Mupirocin resistant MRSA isolates belonged to ST15-SCCmec IV/t084 (40%), ST22-SCCmec IV/t790 (23.3%), ST239-SCCmec III/t631 (20%), and ST239-SCCmec III/t030 (16.7%) clones. The results showed that genetically diverse strains of MRSA are circulating in our burn hospitals with relatively high prevalence of ST239-SCCmec III/t037 clone. The findings support the need for regular surveillance of MRSA to determine the distribution of

  5. Emergence of SCCmec type III with variable antimicrobial resistance profiles and spa types among methicillin-resistant Staphylococcus aureus isolated from healthcare- and community-acquired infections in the west of Iran.

    Science.gov (United States)

    Mohammadi, Sattar; Sekawi, Zamberi; Monjezi, Azam; Maleki, Mohammad-Hossein; Soroush, Setareh; Sadeghifard, Nourkhoda; Pakzad, Iraj; Azizi-Jalilian, Farid; Emaneini, Mohammad; Asadollahi, Khairollah; Pourahmad, Fazel; Zarrilli, Raffaele; Taherikalani, Morovat

    2014-08-01

    Methicillin-resistant Staphylococcus aureus (MRSA) is a pathogen of public health importance. The prevalence of MRSA and its antimicrobial resistance pattern, as well as SCCmec and spa types, remain unclear both in the community and in the hospitals of the western region of Iran. One hundred MRSA isolates were collected from different hospitals in the west of Iran during 2010-2011. Antimicrobial susceptibility testing to 15 antimicrobial agents was carried out by disk agar diffusion (DAD) method in accordance with the Clinical and Laboratory Standards Institute guidelines. Vancomycin minimum inhibitory concentrations (MICs) were evaluated by a broth microdilution method. The Etest was used for the detection of highly gentamicin-resistant MRSA. A combination of single and multiplex PCR was used for the detection of different resistance genes, including beta-lactamase, aminoglycoside modifying enzymes (AMEs), and macrolide-lincosamine, and for SCCmec typing of MRSA isolates. Genotyping of MRSA isolates was performed via spa typing. All tested isolates were susceptible to quinupristin-dalfopristin, linezolid, and vancomycin, but were resistant to penicillin (100%), erythromycin (50%), clindamycin (27%), and gentamicin (18%). MIC50 and MIC90 was 256 μg/ml among gentamicin-resistant MRSA. The most prevalent AME genes among aminoglycoside-resistant isolates were aac(6')-1e-aph(2")-1a (77.8%), aph(3')-IIIa (38.9%), and ant(4')-1a (27.8%). Nearly all tetracycline- and erythromycin-resistant MRSA had ermA and/or ermC but not ermB. Five SCCmec types and subtypes, 13 spa types, and four BURP groups (A-D) were identified. SCCmec types III (45%) and IVc (24%), spa type t701 (30%), and new spa type t12311 (15%) were the most prevalent among MRSA isolates. This study showed the emergence of MRSA with SCCmec type III and with spa types t12311, t10740, t1234, t1991, and t2651 with different phenotypic and genotypic antimicrobial resistance in the west of Iran. We found different

  6. Characterization of Staphylococcus aureus from distinct geographic locations in China: an increasing prevalence of spa-t030 and SCCmec type III.

    Science.gov (United States)

    Chen, Yong; Liu, Zhengxiang; Duo, Libo; Xiong, Jie; Gong, Yanwen; Yang, Jiyong; Wang, Zhanke; Wu, Xuqin; Lu, Zhongyi; Meng, Xiangzhao; Zhao, Jingya; Zhang, Changjian; Wang, Fang; Zhang, Yulong; Zhang, Mengqiang; Han, Li

    2014-01-01

    Staphylococcus aureus belongs to one of the most common bacteria causing healthcare and community associated infections in China, but their molecular characterization has not been well studied. From May 2011 to June 2012, a total of 322 non-duplicate S. aureus isolates were consecutively collected from seven tertiary care hospitals in seven cities with distinct geographical locations in China, including 171 methicillin sensitive S. aureus (MSSA) and 151 MRSA isolates. All isolates were characterized by spa typing. The presence of virulence genes was tested by PCR. MRSA were further characterized by SCCmec typing. Seventy four and 16 spa types were identified among 168 MSSA and 150 MRSA, respectively. One spa type t030 accounted for 80.1% of all MRSA isolates, which was higher than previously reported, while spa-t037 accounted for only 4.0% of all MRSA isolates. The first six spa types (t309, t189, t034, t377, t078 and t091) accounted for about one third of all MSSA isolates. 121 of 151 MRSA isolates (80.1%) were identified as SCCmec type III. pvl gene was found in 32 MSSA (18.7%) and 5 MRSA (3.3%) isolates, with ST22-MSSA-t309 as the most commonly identified strain. Compared with non-epidemic MRSA clones, epidemic MRSA clones (corresponding to ST239) exhibited a lower susceptibility to rifampin, ciprofloxacin, gentamicin and trimethoprim-sulfamethoxazole, a higher prevalence of sea gene and a lower prevalence of seb, sec, seg, sei and tst genes. The increasing prevalence of multidrug resistant spa-t030 MRSA represents a major public health problem in China.

  7. Characterization of Staphylococcus aureus from distinct geographic locations in China: an increasing prevalence of spa-t030 and SCCmec type III.

    Directory of Open Access Journals (Sweden)

    Yong Chen

    Full Text Available Staphylococcus aureus belongs to one of the most common bacteria causing healthcare and community associated infections in China, but their molecular characterization has not been well studied. From May 2011 to June 2012, a total of 322 non-duplicate S. aureus isolates were consecutively collected from seven tertiary care hospitals in seven cities with distinct geographical locations in China, including 171 methicillin sensitive S. aureus (MSSA and 151 MRSA isolates. All isolates were characterized by spa typing. The presence of virulence genes was tested by PCR. MRSA were further characterized by SCCmec typing. Seventy four and 16 spa types were identified among 168 MSSA and 150 MRSA, respectively. One spa type t030 accounted for 80.1% of all MRSA isolates, which was higher than previously reported, while spa-t037 accounted for only 4.0% of all MRSA isolates. The first six spa types (t309, t189, t034, t377, t078 and t091 accounted for about one third of all MSSA isolates. 121 of 151 MRSA isolates (80.1% were identified as SCCmec type III. pvl gene was found in 32 MSSA (18.7% and 5 MRSA (3.3% isolates, with ST22-MSSA-t309 as the most commonly identified strain. Compared with non-epidemic MRSA clones, epidemic MRSA clones (corresponding to ST239 exhibited a lower susceptibility to rifampin, ciprofloxacin, gentamicin and trimethoprim-sulfamethoxazole, a higher prevalence of sea gene and a lower prevalence of seb, sec, seg, sei and tst genes. The increasing prevalence of multidrug resistant spa-t030 MRSA represents a major public health problem in China.

  8. Characterization of a Novel Arginine Catabolic Mobile Element (ACME) and Staphylococcal Chromosomal Cassette mec Composite Island with Significant Homology to Staphylococcus epidermidis ACME type II in Methicillin-Resistant Staphylococcus aureus Genotype ST22-MRSA-IV.

    LENUS (Irish Health Repository)

    Shore, Anna C

    2011-02-22

    The arginine catabolic mobile element (ACME) is prevalent among ST8-MRSA-IVa (USA300) isolates and evidence suggests that ACME enhances the ability of ST8-MRSA-IVa to grow and survive on its host. ACME has been identified in a small number of isolates belonging to other MRSA clones but is widespread among coagulase-negative staphylococci (CoNS). This study reports the first description of ACME in two distinct strains of the pandemic ST22-MRSA-IV clone. A total of 238 MRSA isolates recovered in Ireland between 1971 and 2008 were investigated for ACME using a DNA microarray. Twenty-three isolates (9.7%) were ACME-positive, all were either MRSA genotype ST8-MRSA-IVa (7\\/23, 30%) or ST22-MRSA-IV (16\\/23, 70%). Whole-genome sequencing and comprehensive molecular characterization revealed the presence of a novel 46-kb ACME and SCCmec composite island (ACME\\/SCCmec-CI) in ST22-MRSA-IVh isolates (n = 15). This ACME\\/SCCmec-CI consists of a 12-kb DNA region previously identified in ACME type II in S. epidermidis ATCC 12228, a truncated copy of the J1 region of SCCmec I and a complete SCCmec IVh element. The composite island has a novel genetic organization with ACME located within orfX and SCCmec located downstream of ACME. One pvl-positive ST22-MRSA-IVa isolate carried ACME located downstream of SCCmec IVa as previously described in ST8-MRSA-IVa. These results suggest that ACME has been acquired by ST22-MRSA-IV on two independent occasions. At least one of these instances may have involved horizontal transfer and recombination events between MRSA and CoNS. The presence of ACME may enhance dissemination of ST22-MRSA-IV, an already successful MRSA clone.

  9. Genetics Home Reference: mucopolysaccharidosis type IV

    Science.gov (United States)

    ... types of mucopolysaccharidosis, MPS IV does not affect intelligence. The life expectancy of individuals with MPS IV ... GM1-gangliosidosis and Morquio B patients: possible common origin for the prevalent p.R59H mutation among gypsies. ...

  10. Antimicrobial susceptibility pattern and SCCmec types of methicillin-resistant coagulase-negative staphylococci from subclinical bovine mastitis in Hatay, Turkey

    Directory of Open Access Journals (Sweden)

    Aslantaş Özkan

    2014-12-01

    Full Text Available Eighty-nine isolates of coagulase-negative staphylococci (CoNS of eight species from subclinical bovine mastitis were screened for the phenotypic and genotypic methicilline-resistance. In addition, all methicillin-resistant (MR isolates indicating the mecA gene were examined by PCR for the antimicrobial susceptibility patterns, and staphylococcal cassette chromosome mec (SCCmec types were also determined by multiplex PCR. A total of 21 (23.6% CoNS isolates were found to be resistant to oxacillin in broth microdilution assay. All isolates phenotypically resistant to oxacillin did not have the mecA gene, which was only found in 14.6% (13 of the isolates. Most MR-CoNS isolates were highly resistant to erythromycin (92.3%, fusidic acid (84.6%, penicillin (76.9%, and rifampycin (61.5%, and susceptible to mupirocin (100%, tetracycline (100%, vancomycin (100%, clindamycin (92.3%, and sulfamethoxazole-trimethoprim (69.2%. In conclusion, a high rate of antimicrobial resistance among MR-CoNS isolated from food producing animals emphasises the need for periodic surveillance of their resistance.

  11. Determining the prevalence of SCCmec polymorphism, virulence and antibiotic resistance genes among methicillin-resistant Staphylococcus aureus (MRSA) isolates collected from selected hospitals in west of Iran.

    Science.gov (United States)

    Taherikalani, Morovat; Mohammadzad, Mohammad Reza; Soroush, Setareh; Maleki, Mohammad Hossein; Azizi-Jalilian, Farid; Pakzad, Iraj; Sadeghifard, Nourkhoda; Asadollahi, Parisa; Emaneini, Mohammad; Monjezi, Aazam; Alikhani, Mohammad Yousef

    2016-04-01

    Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most important pathogens worldwide and compared to other staphylococcal species that are associated with higher mortality rate. A total of 500 Staphylococcus spp. was collected from selected hospitals in Ilam, Kermanshah, Khorram Abad and Hamadan cities and, via phenotypic and genotypic methods, was assessed to find MRSA. The presence or absence of prevalent antibiotic resistance genes and virulence genes was evaluated among MRSA isolates, using polymerase chain reaction (PCR) method, and then the SCCmec typing of these isolates was assayed by multiplex PCR. A total of 372 (74.4%) Stapylococcus spp. isolates were identified as S. aureus, among which 200 (53.8%) possessed the mecA gene and were distinguished as MRSA. All of MRSA isolates contained blaZ gene. The frequency of ermA and ermC genes among erythromycin-resistant MRSA isolates was 21.6% and 66.7%, respectively. The frequency of the virulence genes eta, hla and sea among MRSA isolates was 10%, 80.5% and 100%, respectively. SCCmec type IV accounted for 30.6% of the MRSA isolates and SCCmec type III, SCCmec type II and SCCmec type I accounted for 30%, 22% and 17.5% of the isolates, respectively. The antibiotic resistance genes and the virulence genes of blaZ, hla, sea, eta and ermC had high frequencies among the MRSA isolates. This study showed that the antibiotic resistance genes had higher frequencies among SCCmec types I and IV, which confirms the previous reports in this field.

  12. Characterization of a novel arginine catabolic mobile element (ACME) and staphylococcal chromosomal cassette mec composite island with significant homology to Staphylococcus epidermidis ACME type II in methicillin-resistant Staphylococcus aureus genotype ST22-MRSA-IV.

    LENUS (Irish Health Repository)

    Shore, Anna C

    2011-05-01

    The arginine catabolic mobile element (ACME) is prevalent among methicillin-resistant Staphylococcus aureus (MRSA) isolates of sequence type 8 (ST8) and staphylococcal chromosomal cassette mec (SCCmec) type IVa (USA300) (ST8-MRSA-IVa isolates), and evidence suggests that ACME enhances the ability of ST8-MRSA-IVa to grow and survive on its host. ACME has been identified in a small number of isolates belonging to other MRSA clones but is widespread among coagulase-negative staphylococci (CoNS). This study reports the first description of ACME in two distinct strains of the pandemic ST22-MRSA-IV clone. A total of 238 MRSA isolates recovered in Ireland between 1971 and 2008 were investigated for ACME using a DNA microarray. Twenty-three isolates (9.7%) were ACME positive, and all were either MRSA genotype ST8-MRSA-IVa (7\\/23, 30%) or MRSA genotype ST22-MRSA-IV (16\\/23, 70%). Whole-genome sequencing and comprehensive molecular characterization revealed the presence of a novel 46-kb ACME and staphylococcal chromosomal cassette mec (SCCmec) composite island (ACME\\/SCCmec-CI) in ST22-MRSA-IVh isolates (n=15). This ACME\\/SCCmec-CI consists of a 12-kb DNA region previously identified in ACME type II in S. epidermidis ATCC 12228, a truncated copy of the J1 region of SCCmec type I, and a complete SCCmec type IVh element. The composite island has a novel genetic organization, with ACME located within orfX and SCCmec located downstream of ACME. One PVL locus-positive ST22-MRSA-IVa isolate carried ACME located downstream of SCCmec type IVa, as previously described in ST8-MRSA-IVa. These results suggest that ACME has been acquired by ST22-MRSA-IV on two independent occasions. At least one of these instances may have involved horizontal transfer and recombination events between MRSA and CoNS. The presence of ACME may enhance dissemination of ST22-MRSA-IV, an already successful MRSA clone.

  13. Differences between "classical" risk factors for infections caused by methicillin-resistant Staphylococcus aureus (MRSA) and risk factors for nosocomial bloodstream infections caused by multiple clones of the staphylococcal cassette chromosome mec type IV MRSA strain.

    Science.gov (United States)

    Vidal, Paula M; Trindade, Priscila A; Garcia, Tamara O; Pacheco, Renata L; Costa, Silvia F; Reinert, Cristina; Hiramatsu, Keiichi; Mamizuka, Elsa M; Garcia, Cilmara P; Levin, Anna S

    2009-02-01

    To identify risk factors associated with nosocomial bloodstream infections caused by multiple clones of the staphylococcal cassette chromosome mec (SCCmec) type IV strain of methicillin-resistant Staphylococcus aureus (MRSA). An unmatched case-control study (at a ratio of 1:2) performed during the period from October 2002 through September 2003. A 2,000-bed tertiary care teaching hospital affiliated with the University of São Paulo in São Paulo, Brazil. Case patients (n=30) were defined either as patients who had a bloodstream infection due to SCCmec type IV MRSA diagnosed at least 48 hours after hospital admission or as neonates with the infection who were born in the hospital. Control patients (n=60) were defined as patients with SCCmec type III MRSA infection diagnosed at least 48 hours after hospital admission. Genes encoding virulence factors were studied in the isolates recovered from case patients, and molecular typing of the SCCmec type IV MRSA isolates was also done by pulsed-field gel electrophoresis and multilocus sequence typing. In multivariate analysis, the following 3 variables were significantly associated with having a nosocomial bloodstream infection caused by SCCmec type IV strains of MRSA: an age of less than 1 year, less frequent use of a central venous catheter (odds ratio [OR], 0.07 [95% confidence interval {CI}, 0.02-0.28]; p= .025), and female sex. A second analysis was performed that excluded the case and control patients from the neonatal unit, and, in multivariate analysis, the following variables were significantly associated with having a nosocomial bloodstream infection caused by SCCmec type IV strains of MRSA: less frequent use of a central venous catheter (OR, 0.12 [95% CI, 0.03-0.55]; p= .007), lower Acute Physiology and Chronic Health Evaluation II score on admission (OR, 0.14 [95% CI, 0.03-0.61]; p= .009), less frequent surgery (OR, 0.21 [95% CI, 0.06-0.83]; p= .025), and female sex (OR, 5.70 [95% CI, 1.32-24.66]; p= .020). Of

  14. Enhanced discrimination of highly clonal ST22-methicillin-resistant Staphylococcus aureus IV isolates achieved by combining spa, dru, and pulsed-field gel electrophoresis typing data.

    LENUS (Irish Health Repository)

    Shore, Anna C

    2010-05-01

    ST22-methicillin-resistant Staphylococcus aureus type IV (ST22-MRSA-IV) is endemic in Irish hospitals and is designated antibiogram-resistogram type-pulsed-field group (AR-PFG) 06-01. Isolates of this highly clonal strain exhibit limited numbers of pulsed-field gel electrophoresis (PFGE) patterns and spa types. This study investigated whether combining PFGE and spa typing with DNA sequencing of the staphylococcal cassette chromosome mec element (SCCmec)-associated direct repeat unit (dru typing) would improve isolate discrimination. A total of 173 MRSA isolates recovered in one Irish hospital during periods in 2007 and 2008 were investigated using antibiogram-resistogram (AR), PFGE, spa, dru, and SCCmec typing. Isolates representative of each of the 17 pulsed-field group 01 (PFG-01) spa types identified underwent multilocus sequence typing, and all isolates were ST22. Ninety-seven percent of isolates (168 of 173) exhibited AR-PFG 06-01 or closely related AR patterns, and 163 of these isolates harbored SCCmec type IVh. The combination of PFGE, spa, and dru typing methods significantly improved discrimination of the 168 PFG-01 isolates, yielding 65 type combinations with a Simpson\\'s index of diversity (SID) of 96.53, compared to (i) pairwise combinations of spa and dru typing, spa and PFGE typing, and dru and PFGE typing, which yielded 37, 44, and 43 type combinations with SIDs of 90.84, 91.00, and 93.57, respectively, or (ii) individual spa, dru, and PFGE typing methods, which yielded 17, 17, and 21 types with SIDs of 66.9, 77.83, and 81.34, respectively. Analysis of epidemiological information for a subset of PFG-01 isolates validated the relationships inferred using combined PFGE, spa, and dru typing data. This approach significantly enhances discrimination of ST22-MRSA-IV isolates and could be applied to epidemiological investigations of other highly clonal MRSA strains.

  15. Mirizzi Syndrome Type IV: A challenging diagnosis

    Directory of Open Access Journals (Sweden)

    Daniel Navarini

    2016-05-01

    Full Text Available Mirizzi Syndrome type IV is an extremely rare condition, which is confused with the diagnosis of cholangiocarcinoma in many cases. This report describes a case of a forty-three-year old patient, who was forwarded to our department of general surgery with a high suspicion of a choledochal neoplasic lesion. During the hospitalization he was diagnosed with Mirizzi Syndrome type IV. We concisely describe the case and the literature review about this pathology.

  16. Observational properties of decameter type IV bursts

    Science.gov (United States)

    Melnik, Valentin; Brazhenko, Anatoly; Rucker, Helmut; Konovalenko, Alexander; Briand, Carine; Dorovskyy, Vladimir; Zarka, Philippe; Frantzusenko, Anatoly; Panchenko, Michael; Poedts, Stefan; Zaqarashvili, Teimuraz; Shergelashvili, Bidzina

    2013-04-01

    Oscillations of decameter type IV bursts were registered during observations of solar radio emission by UTR-2, URAN-2 and NDA in 2011-2012. Large majority of these bursts were accompanied by coronal mass ejections (CMEs), which were observed by SOHO and STEREO in the visible light. Only in some cases decameter type IV bursts were not associated with CMEs. The largest periods of oscillations P were some tens of minutes. There were some modes of long periods of oscillations simultaneously. Periods of oscillations in flux and in polarization profiles were close. Detailed properties of oscillations at different frequencies were analyzed on the example of two type IV bursts. One of them was observed on April 7, 2011 when a CME happened. Another one (August 1, 2011) was registered without any CME. The 7 April type IV burst had two periods in the frames 75-85 and 35-85 minutes. Interesting feature of these oscillations is decreasing periods with time. The observed decreasing rates dP/dt equaled 0.03-0.07. Concerning type IV burst observed on August 1, 2011 the period of its oscillations increases from 17 min. at 30 MHz to 44 min. at 10 MHz. Connection of type IV burst oscillations with oscillations of magnetic arches and CMEs at corresponding altitudes are discussed. The work is fulfilled in the frame of FP7 project "SOLSPANET".

  17. Type IV Wind Turbine Model

    DEFF Research Database (Denmark)

    Hansen, Anca Daniela; Margaris, Ioannis D.

    project to be incorporated in the wind power plant level. This document describes the Type 4 wind turbine simulation model, implemented in the EaseWind project. The implemented wind turbine model is one of the initial necessary steps toward integrating new control services in the wind power plant level....... In the project, this wind turbine model will be further incorporated in a wind power plant model together with the implementation in the wind power control level of the new control functionalities (inertial response, synchronising power and power system damping). For this purpose an aggregate wind power plant...... (WPP) will be considered. The aggregate WPP model, which will be based on the upscaling of the individual wind turbine model on the electrical part, will make use of an equivalent wind speed. The implemented model follows the basic structure of the generic standard Type 4 wind turbine model proposed...

  18. Muscle Function in Osteogenesis Imperfecta Type IV.

    Science.gov (United States)

    Veilleux, Louis-Nicolas; Darsaklis, Vasiliki B; Montpetit, Kathleen; Glorieux, Francis H; Rauch, Frank

    2017-05-04

    Results of previous studies suggest that children and adolescents with osteogenesis imperfecta (OI) type IV have muscle force deficits. However, muscle function remains to be objectively quantified in this population. This study aimed to assess upper and lower extremity muscle function in patients with OI type IV. It was carried out in the outpatient department of a pediatric orthopedic hospital; 27 individuals with OI type IV (7-21 years; 13 males), 27 age- and sex-matched individuals with OI type I, and 27 age- and sex-matched controls. Upper extremity muscle force was assessed with hydraulic hand dynamometry, and lower extremity muscle function (peak force per body weight and peak power per body mass) was measured by mechanography through five tests: multiple two-legged hopping, multiple one-legged hopping, single two-legged jump, chair-rise test, and heel-rise test. Upper-limb grip force was normal for patients with OI type IV when compared to height and sex reference data (average z-score = 0.17 ± 1.30; P = 0.88). Compared to age- and sex-matched controls, patients with OI type IV had approximately 30% lower-limb peak force and 50% peak power deficits (P values <0.05). At the lower-limb level, they had a 50% lower peak power than age- and sex-matched patients with OI type I (P < 0.05). Patients with OI type IV have normal upper-limb muscle force but a muscle function deficit at the lower-limb level. These results suggest that lower-limb muscle weakness may contribute to functional deficits in these individuals.

  19. Comparative Genotypes, Staphylococcal Cassette Chromosome mec (SCCmec) Genes and Antimicrobial Resistance amongst Staphylococcus epidermidis and Staphylococcus haemolyticus Isolates from Infections in Humans and Companion Animals.

    Science.gov (United States)

    McManus, Brenda A; Coleman, David C; Deasy, Emily C; Brennan, Gráinne I; O' Connell, Brian; Monecke, Stefan; Ehricht, Ralf; Leggett, Bernadette; Leonard, Nola; Shore, Anna C

    2015-01-01

    This study compares the characteristics of Staphylococcus epidermidis (SE) and Staphylococcus haemolyticus (SH) isolates from epidemiologically unrelated infections in humans (Hu) (28 SE-Hu; 8 SH-Hu) and companion animals (CpA) (12 SE-CpA; 13 SH-CpA). All isolates underwent antimicrobial susceptibility testing, multilocus sequence typing and DNA microarray profiling to detect antimicrobial resistance and SCCmec-associated genes. All methicillin-resistant (MR) isolates (33/40 SE, 20/21 SH) underwent dru and mecA allele typing. Isolates were predominantly assigned to sequence types (STs) within a single clonal complex (CC2, SE, 84.8%; CC1, SH, 95.2%). SCCmec IV predominated among MRSE with ST2-MRSE-IVc common to both Hu (40.9%) and CpA (54.5%). Identical mecA alleles and nontypeable dru types (dts) were identified in one ST2-MRSE-IVc Hu and CpA isolate, however, all mecA alleles and 2/4 dts detected among 18 ST2-MRSE-IVc isolates were closely related, sharing >96.5% DNA sequence homology. Although only one ST-SCCmec type combination (ST1 with a non-typeable [NT] SCCmec NT9 [class C mec and ccrB4]) was common to four MRSH-Hu and one MRSH-CpA, all MRSH isolates were closely related based on similar STs, SCCmec genes (V/VT or components thereof), mecA alleles and dts. Overall, 39.6% of MR isolates harbored NT SCCmec elements, and ACME was more common amongst MRSE and CpA isolates. Multidrug resistance (MDR) was detected among 96.7% of isolates but they differed in the prevalence of specific macrolide, aminoglycoside and trimethoprim resistance genes amongst SE and SH isolates. Ciprofloxacin, rifampicin, chloramphenicol [fexA, cat-pC221], tetracycline [tet(K)], aminoglycosides [aadD, aphA3] and fusidic acid [fusB] resistance was significantly more common amongst CpA isolates. SE and SH isolates causing infections in Hu and CpA hosts belong predominantly to STs within a single lineage, harboring similar but variable SCCmec genes, mecA alleles and dts. Host and

  20. Ehlers-Danlos syndrome type IV

    Directory of Open Access Journals (Sweden)

    Germain Dominique P

    2007-07-01

    Full Text Available Abstract Ehlers-Danlos syndrome type IV, the vascular type of Ehlers-Danlos syndromes (EDS, is an inherited connective tissue disorder defined by characteristic facial features (acrogeria in most patients, translucent skin with highly visible subcutaneous vessels on the trunk and lower back, easy bruising, and severe arterial, digestive and uterine complications, which are rarely, if at all, observed in the other forms of EDS. The estimated prevalence for all EDS varies between 1/10,000 and 1/25,000, EDS type IV representing approximately 5 to 10% of cases. The vascular complications may affect all anatomical areas, with a tendency toward arteries of large and medium diameter. Dissections of the vertebral arteries and the carotids in their extra- and intra-cranial segments (carotid-cavernous fistulae are typical. There is a high risk of recurrent colonic perforations. Pregnancy increases the likelihood of a uterine or vascular rupture. EDS type IV is inherited as an autosomal dominant trait that is caused by mutations in the COL3A1 gene coding for type III procollagen. Diagnosis is based on clinical signs, non-invasive imaging, and the identification of a mutation of the COL3A1 gene. In childhood, coagulation disorders and Silverman's syndrome are the main differential diagnoses; in adulthood, the differential diagnosis includes other Ehlers-Danlos syndromes, Marfan syndrome and Loeys-Dietz syndrome. Prenatal diagnosis can be considered in families where the mutation is known. Choriocentesis or amniocentesis, however, may entail risk for the pregnant woman. In the absence of specific treatment for EDS type IV, medical intervention should be focused on symptomatic treatment and prophylactic measures. Arterial, digestive or uterine complications require immediate hospitalisation, observation in an intensive care unit. Invasive imaging techniques are contraindicated. Conservative approach is usually recommended when caring for a vascular

  1. Ehlers-Danlos syndrome type IV

    Science.gov (United States)

    Germain, Dominique P

    2007-01-01

    Ehlers-Danlos syndrome type IV, the vascular type of Ehlers-Danlos syndromes (EDS), is an inherited connective tissue disorder defined by characteristic facial features (acrogeria) in most patients, translucent skin with highly visible subcutaneous vessels on the trunk and lower back, easy bruising, and severe arterial, digestive and uterine complications, which are rarely, if at all, observed in the other forms of EDS. The estimated prevalence for all EDS varies between 1/10,000 and 1/25,000, EDS type IV representing approximately 5 to 10% of cases. The vascular complications may affect all anatomical areas, with a tendency toward arteries of large and medium diameter. Dissections of the vertebral arteries and the carotids in their extra- and intra-cranial segments (carotid-cavernous fistulae) are typical. There is a high risk of recurrent colonic perforations. Pregnancy increases the likelihood of a uterine or vascular rupture. EDS type IV is inherited as an autosomal dominant trait that is caused by mutations in the COL3A1 gene coding for type III procollagen. Diagnosis is based on clinical signs, non-invasive imaging, and the identification of a mutation of the COL3A1 gene. In childhood, coagulation disorders and Silverman's syndrome are the main differential diagnoses; in adulthood, the differential diagnosis includes other Ehlers-Danlos syndromes, Marfan syndrome and Loeys-Dietz syndrome. Prenatal diagnosis can be considered in families where the mutation is known. Choriocentesis or amniocentesis, however, may entail risk for the pregnant woman. In the absence of specific treatment for EDS type IV, medical intervention should be focused on symptomatic treatment and prophylactic measures. Arterial, digestive or uterine complications require immediate hospitalisation, observation in an intensive care unit. Invasive imaging techniques are contraindicated. Conservative approach is usually recommended when caring for a vascular complication in a patient suffering

  2. Reconstruction for Type IV Radial Polydactyly.

    Science.gov (United States)

    Wall, Lindley B; Goldfarb, Charles A

    2015-09-01

    Type IV radial polydactyly represents a thumb with an extra proximal and distal phalanx. Assessment of the thumb for surgical reconstruction includes observing thumb function, evaluating thumb size and stability, and assessing the first web space. Reconstruction includes excision of the smaller thumb, typically the radial thumb, and re-creating thumb stability and alignment by addressing tendon insertion and joint orientation. Although surgical results are satisfying and complications are uncommon, additional surgical intervention may be required over time owing to thumb malalignment or instability. Copyright © 2015 American Society for Surgery of the Hand. Published by Elsevier Inc. All rights reserved.

  3. Genetics Home Reference: mucolipidosis type IV

    Science.gov (United States)

    ... it does result in unusually high levels of gastrin in the blood. Gastrin is a hormone that regulates the production of ... Resources (2 links) GeneReview: Mucolipidosis IV MedlinePlus Encyclopedia: Gastrin General Information from MedlinePlus (5 links) Diagnostic Tests ...

  4. On the Bivariate Kummer-Beta Type IV Distribution

    NARCIS (Netherlands)

    Jacobs, R.; Bekker, A.; Human, S.W.

    2012-01-01

    In this article, the non central bivariate Kummer-beta Type IV distribution is introduced and derived via the Laplace transform of the non central bivariate beta distribution by Gupta et al. (2011 ). We focus on and discuss the central bivariate Kummer-beta Type IV distribution; this distribution is

  5. Hepatocellular carcinoma in glycogen storage disease type IV

    OpenAIRE

    de Moor, R A; Schweizer, J.; Van Hoek, B.; Wasser, M.; Vink, R.; Maaswinkel-Mooy, P.

    2000-01-01

    A 13 year old patient with juvenile type IV glycogen storage disease died of the complications of hepatocellular carcinoma. To our knowledge this is the first reported case of hepatocellular carcinoma in association with type IV glycogen storage disease.



  6. Genetics Home Reference: glycogen storage disease type IV

    Science.gov (United States)

    ... hepatic type of GSD IV often die of liver failure in early childhood. The non-progressive hepatic type ... and Advocacy Resources (7 links) American Heart Association: Pediatric Cardiomyopathies American Liver Foundation Association for Glycogen Storage Disease CLIMB: Children ...

  7. Simultaneous expression of 70 kilodalton type IV collagenase and type IV collagen alpha 1 (IV) chain genes by cells of early human placenta and gestational endometrium.

    Science.gov (United States)

    Autio-Harmainen, H; Hurskainen, T; Niskasaari, K; Höyhtyä, M; Tryggvason, K

    1992-08-01

    In this study we used in situ hybridization to investigate the expression of the genes 70 kilodalton (kd) collagenase and the alpha 1(IV) collagen chain of type IV collagen in cells of early human placenta and gestational endometrium. The aim was to study the spatial distribution of these gene expressions within a developing tissue which possesses physiologic invasive potential. The results obtained for the 70 kd type IV collagenase mRNA expression were also compared with the immunohistochemical distribution of the corresponding antigen. Expression of mRNAs for these proteins was found in cells of trophoblastic columns, stromal cells of villi and in cells of decidua and endometrial stroma. The only differences between the expressions was the lower level of signals for 70 kd type IV collagenase in fibroblastic stromal cells and endothelial cells of villi and in the pericytic cells of spiral arteries. Otherwise the results for both types of mRNA were comparable. We also studied the immunohistochemical distribution of the 70 kd type IV collagenase using specific monoclonal antibodies against the enzyme. Immunohistochemistry supported well the findings obtained by in situ hybridization. The results indicate that the genes for the 70 kd type IV collagenase and for the alpha 1(IV) collagen chain are simultaneously active in cells of placenta and gestational endometrium and the same cells which produce type IV collagen also can produce the cleaving enzyme, the 70 kd type IV collagenase. The results also show that the cytotrophoblastic cells, which during early pregnancy invade the extracellular matrix and spiral arteries of uterine wall contain significant amount of mRNA for the 70 kd type IV collagenase. This finding supports the concept that the 70 kd type IV collagenase would be important for invasion, and in the case of this study, also for the physiologic invasion of placental cytotrophoblasts.

  8. A case of hereditary sensory autonomic neuropathy type IV

    Directory of Open Access Journals (Sweden)

    G P Prashanth

    2012-01-01

    Full Text Available Hereditary sensory autonomic neuropathy type IV (HSAN -IV, also known as congenital insensitivity to pain with anhidrosis, is a very rare condition that presents in infancy with anhidrosis, absence of pain sensation and self -mutilation. Developmental delay and mental retardation are usually present. Ultrastructural study of the peripheral nerves demonstrates loss of the unmyelinated and small myelinated fibers. We here report a 8 year -old boy with HSAN IV with typical clinical features where the diagnosis was supported by nerve biopsy findings. However, our case was unusual since mental development was normal.

  9. CASE REPORT Dual (type IV) left anterior descending artery

    African Journals Online (AJOL)

    [1] Type IV can be differentiated from the other types by the long LAD originating from the RCA. Case report. A 52-year-old man was referred to our hospital for chest pain during exercise. The physical examination and laboratory findings were unremarkable. The patient underwent coronary CT angiography (CTA).

  10. Distribution of SCCmec-associated phenol-soluble modulin in staphylococci.

    LENUS (Irish Health Repository)

    Monecke, Stefan

    2012-04-01

    The recently described phenol-soluble modulin PSM-mec was detected in Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus fleuretti, Staphylococcus hominis, Staphylococcus pseudintermedius, Staphylococcus saprophyticus, Staphylococcus simulans and Staphylococcus vitulinus from different hosts (humans, goats, dogs, cats, pigs, cattle and turkeys). It was identified in isolates harbouring SCCmec types II, IIA, IIB, IID, III, VIII and in some irregular or truncated elements.

  11. Evaluation of double locus (clfB and spa) sequence typing for studying molecular epidemiology of methicillin-resistant Staphylococcus aureus in Taiwan.

    Science.gov (United States)

    Huang, Chen-Cheng; Ho, Cheng-Mao; Chen, Hui-Chen; Li, Chi-Yuan; Tien, Ni; Fan, Hsiu-Mei; Ge, Mao-Cheng; Lu, Jang-Jih

    2017-10-01

    Pulsed-field gel electrophoresis (PFGE) is the "gold standard" for epidemiological investigation of methicillin-resistant Staphylococcus aureus (MRSA), but several DNA sequence-based methods have been developed in MRSA typing because of the unambiguous results. Ninety-one MRSA isolates were collected from the blood cultures of different patients from July 2008 to December 2008 in central Taiwan. The molecular characteristics of each isolate, including double locus sequence typing (DLST; spa and clfB typing), Staphylococcus cassette chromosome mec (SCCmec), and PFGE were determined for comparison. Five major clfB types (types A-E), 18 spa types, 33 DLST genotypes, five SCCmec types, 17 pulsotypes have been observed. Three major DLST genotypes (A1-t002, C0-t037, and B1-t437) and two major pulsotypes (6 and 8) were identified. Most clfB type A isolates (97.1%) were SCCmec type II and all clfB type C isolates (100%) were SCCmec type III. Most clfB type B isolates (88.9%) were SCCmec type IV (59.3%) and VT (29.6%). All (100%) clfB subtypes A1, A2, and C isolates and 70.4% of clfB type B isolates belonged to healthcare-associated-MRSA. The average congruence was 57.7% between DLST and PFGE, and 96.6% between clfB and SCCmec type. The index of discrimination of SCCmec, clfB, spa, PFGE, and DLST was 0.72, 0.79, 0.80, 0.81, and 0.87, respectively. ClfB type has high congruence with SCCmec type. The DLST method in this study yielded a higher discriminatory power than PFGE in local investigation of molecular epidemiology of MRSA and a promising alternative to PFGE. Copyright © 2015. Published by Elsevier B.V.

  12. Osteogenesis imperfecta Type IV: a newly identified variant at ...

    African Journals Online (AJOL)

    Osteogenesis imperfecta is a clinically heterogenous disease caused by defective collagen syntesis associated with a mutation in the COL1A1 or COL1A2 genes. In this report, we present a case of osteogenesis imperfecta (OI) type IV, seen in a female fetus with incurved femurs at 18 weeks of gestation. Molecular analysis ...

  13. Biofilm formation by Pseudomonas aeruginosa wild type, flagella and type IV pili mutants

    DEFF Research Database (Denmark)

    Klausen, M.; Heydorn, Arne; Ragas, Paula Cornelia

    2003-01-01

    Biofilm formation by Gfp-tagged Pseudomonas aeruginosa PAO1 wild type, flagella and type IV pili mutants in flow chambers irrigated with citrate minimal medium was characterized by the use of confocal laser scanning microscopy and comstat image analysis. Flagella and type IV pili were not necessary...... for P. aeruginosa initial attachment or biofilm formation, but the cell appendages had roles in biofilm development, as wild type, flagella and type IV pili mutants formed biofilms with different structures. Dynamics and selection during biofilm formation were investigated by tagging the wild type...... and flagella/type IV mutants with Yfp and Cfp and performing time-lapse confocal laser scanning microscopy in mixed colour biofilms. The initial microcolony formation occurred by clonal growth, after which wild-type P. aeruginosa bacteria spread over the substratum by means of twitching motility. The wild-type...

  14. Outcome of surgical treatment of type IV capitellum frac-

    Directory of Open Access Journals (Sweden)

    Singh Ajay Pal

    2012-08-01

    Full Text Available 【Abstract】 Objective: Fractures of the capitellum and trochlea constitute less than 1% of all elbow fractures and a shear fracture involving the capitellum and extending medially into most of the trochlea is rarely reported. Type IV capitellum fracture is still controversial in regard to its ra-diographic appearance, surgical approach and osteosynthesis. We report 10 cases of type IV capitellum fracture with a view to elucidating its clinical features and treatment outcome. Methods: We treated 10 patients of type IV capitellum fracture with a mean age of 32 years. A uniform surgical approach and postoperative rehabilitation were followed. Results: Nine patients presented to us after a mean of 4 days of injury and one patient was nonunion after 6 months of injury who had been treated conservatively by a bone setter. Double arc sign was absent in 6 cases. Intraopera-tively 6 capitellotrochlear fragments were devoid of soft Chin J Traumatol 2012;15(4:201-205 DOI: 10.3760/cma.j.issn.1008-1275.2012.04.002 Punjab Civil Medical Services-I Mukerian, Punjab, In-dia (Singh AP Department of Orthopaedics, UCMS & GTB Hospital, Delhi, India (Dhammi IK and Garg V Swami Premanand Hospital, Mukerian, Punjab, India (Singh AP *Corresponding author: Tel: 98-72069734, Email: docajaypal@gmail.com C oronal shear fractures of distal end of humeral articular surface involve the capitellum and trochlea. These fractures are difficult to assess accurately on plain radiographs and the limited amount of subchondral bone available for stable internal fixation makes the operative treatment equally difficult. 1 Joint stiffness, instability and osteoarthrosis are complications resulting from treatment failures. 2 The major classification systems are proposed by Bryan and Dubberley et al. 3,4 Type IV fracture is a fracture involving the capitellum and extends to more than lateral half of the trochlea. 1 We report an analysis of ten cases of type IV capi-tissue attachments. By

  15. Spinal type IV arteriovenous malformations (perimedullary fistulas) in children.

    Science.gov (United States)

    Sure, U; Wakat, J P; Gatscher, S; Becker, R; Bien, S; Bertalanffy, H

    2000-08-01

    Intradural perimedullary spinal arteriovenous fistula (AVF) is a synonym for type IV spinal arteriovenous malformation (AVM). It is an important clinical differential diagnosis in all patients with slowly progressive or acute spinal symptoms. Perimedullary AVFs are rare in the paediatric age group. We report the treatment regimen and the clinico-radiological findings for these malformations at our institution. Of four paediatric patients, three individuals with persistent fistulas after endovascular treatment were operated on. The malformations were obliterated completely in all patients without any morbidity. In three patients previously documented neurological symptoms resolved during follow-up. If slowly progressive or acute radicular or medullary symptoms arise in children, a spinal arteriovenous malformation should be ruled out by MRI. A combined endovascular and surgical treatment of paediatric spinal AVM type IV (perimedullary AVFs) carries a low risk of morbidity and is reasonably effective.

  16. Sequence Domain Harmonic Modeling of Type-IV Wind Turbines

    DEFF Research Database (Denmark)

    Guest, Emerson; Jensen, Kim Høj; Rasmussen, Tonny Wederberg

    2017-01-01

    A sequence domain (SD) harmonic model of a gridconnected voltage-source converter is developed for decoupling converter generated voltage harmonics from voltage harmonics in the external grid. The modeling procedure includes a derivation of the baseband frequency response for regular......-sampled pulsewidth modulation and an analysis of converter generated voltage harmonics due to compensated dead-time. The decoupling capabilities of the proposed the SD harmonic model are verified through a power quality (PQ) assessment of a 3MW Type-IV wind turbine. The assessment shows that the magnitude and phase...... of low-order odd converter generated voltage harmonics are dependent on the converter operating point and the phase of the fundamental component of converter current respectively. The SD harmonic model can be used to make PQ assessments of Type-IV wind turbines or incorporated into harmonic load flows...

  17. Genotyping of methicillin-resistant Staphylococcus aureus in the Sultan Qaboos University Hospital, Oman reveals the dominance of Panton–Valentine leucocidin-negative ST6-IV/t304 clone

    Directory of Open Access Journals (Sweden)

    E.E. Udo

    2014-07-01

    Full Text Available The objective of this study was to determine the prevalence and distribution of methicillin-resistant Staphylococcus aureus (MRSA genotypes circulating at a tertiary hospital in the Sultanate of Oman. A total of 79 MRSA isolates were obtained from different clinical samples and investigated using antibiogram, pulsed-field gel electrophoresis (PFGE, staphylococcal chromosome cassette mec (SCCmec, Spa typing and multilocus sequence typing (MLST. The isolates were susceptible to linezolid, vancomycin, teicoplanin, tigecycline and mupirocin but were resistant to tetracycline (30.4%, erythromycin (26.6%, clindamycin (24.1%, trimethoprim (19.0%, ciprofloxacin (17.7%, fusidic acid (15.2% and gentamicin (12.7%. Molecular typing revealed 19 PFGE patterns, 26 Spa types and 21 sequence types. SCCmec-IV (86.0% was the dominant SCCmec type, followed by SCCmec-V (10.1%. SCCmec-III (2.5% and SCCmec-II (1.3% were less common. ST6-IV/t304 (n = 30 and ST1295-IV/t690 (n = 12 were the dominant genotypes followed by ST772-V/t657 (n = 5, ST30-IV/t019/t021 (n = 5, ST22-IV/t852 (n = 4, ST80-IV/t044 (n = 3 and 18 single genotypes that were isolated sporadically. On the basis of SCCmec typing and MLST, 91.2% of the isolates were classified as community-associated MRSA and 8.8% of the isolates (consisting of four ST22-IV/t852, one ST239-III/t632, one ST5-III/t311 and one ST5-II/t003 were classified as healthcare-associated MRSA. The study has revealed the dominance of a Panton–Valentine leucocidin-negative ST6-IV/t304 clone and provided insights into the distribution of antibiotic resistance in MRSA at the tertiary hospital in Oman. It also highlights the importance of surveillance in detecting the emergence of new MRSA clones in a healthcare facility.

  18. Audiological findings in children with mucopolysaccharidoses type i-iv.

    Science.gov (United States)

    Vargas-Gamarra, María F; de Paula-Vernetta, Carlos; Vitoria Miñana, Isidro; Ibañez-Alcañiz, Isabel; Cavallé-Garrido, Laura; Alamar-Velazquez, Agustín

    The aim of our study is to reflect hearing impairment of 23children diagnosed with mucopolysaccharidosis (MPS) typeI, II, III and IV. Retrospective study of the clinical, audiological and treatment (medical vs surgical) findings of 23children diagnosed with MPS typeI, II, III or IV followed at a Tertiary Referral Hospital between 1997 and 2015. Six cases of MPSI, 8 of MPSII, 4 of MPSIII and 5 of MPSIV were reviewed. 71.2% of patients had secretory otitis media (SOM) and 54% of patients had some type of hearing loss (HL). The behaviour of hearing loss was variable in each of the subgroups of MPS, finding greater involvement and variability in typesI and II. Children with MPS have a high risk of hearing loss. A significant percentage of transmissive HL progressing to mixed or sensorineural HL was observed. This was more common in typesI and II. Periodic follow up of these patients is mandatory because of hearing impairment and consequences for their development and quality of life. Copyright © 2016 Elsevier España, S.L.U. and Sociedad Española de Otorrinolaringología y Cirugía de Cabeza y Cuello. All rights reserved.

  19. Association of systemic collagen type IV formation with survival among patients undergoing hemodialysis

    DEFF Research Database (Denmark)

    Leeming, Diana Julie; Karsdal, Morten A; Rasmussen, Lars M

    2013-01-01

    The 7S domain of collagen type IV (P4NP_7S) assessed in plasma represents systemic collagen type IV formation. The objective of the study was to investigate the association of systemic collagen type IV formation with survival among patients undergoing hemodialysis.......The 7S domain of collagen type IV (P4NP_7S) assessed in plasma represents systemic collagen type IV formation. The objective of the study was to investigate the association of systemic collagen type IV formation with survival among patients undergoing hemodialysis....

  20. Palmar spiny keratoderma associated with type IV hyperlipoproteinemia.

    Science.gov (United States)

    Urbani, C E; Moneghini, L

    1998-05-01

    We report the case of a 35-year-old man with acquired palmar spiny keratoderma (SK) in association with type IV hyperlipoproteinemia. Cutaneous lesions appeared 11 years before with no history of any previous skin conditions, topical medication or systemic treatment with Vitamin A-derived drugs, x-irradiation, arsenic poisoning or prolonged sun exposure. Family aggregation for SK or other disorders of keratinization was ruled out. Blood chemistries demonstrated hypertriglyceridemia (422 mg/dl) and elevation in plasma concentration of the pre-beta fraction (VLDL) of lipoproteins pattern compatible with a type IV hyperlipoproteinemia. Family aggregation for this metabolic defect was then confirmed. The histologic hallmark of the 'spine' lesion was a compact column of hyperparakeratotic cells (columnar hyperparakeratosis) continuous with a hypogranular layer, without further evidence of dyskeratotic or vacuolated keratinocytes and inflammatory cells in the corresponding dermis. SK represents a well-definite entity which fulfills precise clinico-histologic criteria. However, three main questions are related to SK: differential diagnosis with porokeratosis on histologic examination (columnar hyperparakeratosis or hyperorthokeratosis alone in SK vs. cornoid lamella accompanied by remarkable dermoepidermal changes in porokeratosis); classificative scheme (proper nomenclature to avoid misleading and confounding names); and nosological arrangement (probably two subsets exist: hereditary or benign; and acquired, or idiopathic, which may be paraneoplastic in about 50% of the patients). To the best of our knowledge this is the first case reporting the association between SK and a metabolic impairment although the real connections linking these conditions are still unclear.

  1. Named entity recognition for bacterial Type IV secretion systems.

    Directory of Open Access Journals (Sweden)

    Sophia Ananiadou

    2011-03-01

    Full Text Available Research on specialized biological systems is often hampered by a lack of consistent terminology, especially across species. In bacterial Type IV secretion systems genes within one set of orthologs may have over a dozen different names. Classifying research publications based on biological processes, cellular components, molecular functions, and microorganism species should improve the precision and recall of literature searches allowing researchers to keep up with the exponentially growing literature, through resources such as the Pathosystems Resource Integration Center (PATRIC, patricbrc.org. We developed named entity recognition (NER tools for four entities related to Type IV secretion systems: 1 bacteria names, 2 biological processes, 3 molecular functions, and 4 cellular components. These four entities are important to pathogenesis and virulence research but have received less attention than other entities, e.g., genes and proteins. Based on an annotated corpus, large domain terminological resources, and machine learning techniques, we developed recognizers for these entities. High accuracy rates (>80% are achieved for bacteria, biological processes, and molecular function. Contrastive experiments highlighted the effectiveness of alternate recognition strategies; results of term extraction on contrasting document sets demonstrated the utility of these classes for identifying T4SS-related documents.

  2. Identification of Anaplasma marginale type IV secretion system effector proteins.

    Directory of Open Access Journals (Sweden)

    Svetlana Lockwood

    Full Text Available Anaplasma marginale, an obligate intracellular alphaproteobacterium in the order Rickettsiales, is a tick-borne pathogen and the leading cause of anaplasmosis in cattle worldwide. Complete genome sequencing of A. marginale revealed that it has a type IV secretion system (T4SS. The T4SS is one of seven known types of secretion systems utilized by bacteria, with the type III and IV secretion systems particularly prevalent among pathogenic Gram-negative bacteria. The T4SS is predicted to play an important role in the invasion and pathogenesis of A. marginale by translocating effector proteins across its membrane into eukaryotic target cells. However, T4SS effector proteins have not been identified and tested in the laboratory until now.By combining computational methods with phylogenetic analysis and sequence identity searches, we identified a subset of potential T4SS effectors in A. marginale strain St. Maries and chose six for laboratory testing. Four (AM185, AM470, AM705 [AnkA], and AM1141 of these six proteins were translocated in a T4SS-dependent manner using Legionella pneumophila as a reporter system.The algorithm employed to find T4SS effector proteins in A. marginale identified four such proteins that were verified by laboratory testing. L. pneumophila was shown to work as a model system for A. marginale and thus can be used as a screening tool for A. marginale effector proteins. The first T4SS effector proteins for A. marginale have been identified in this work.

  3. Severe type IV hypersensitivity to 'black henna' tattoo.

    Science.gov (United States)

    Vasilakis, Vasileios; Knight, Bernice; Lidder, Satnam; Frankton, Sarah

    2010-10-06

    A 16-year-old Bangladeshi girl presented with a 9-day history of an extensive pruritic, erythematous, papulovesicular skin eruption to both forearms. Appearance was 5 days following application of a home-made henna preparation. Examination revealed ulceration and scabbing along the whole henna pattern and early keloid formation. A diagnosis of type IV delayed hypersensitivity reaction superimposed by infection was initially made. As in this case, home-made henna preparations commonly combine commercial henna with black hair dye, paraphenylenediamine (PPD). PPD, widely known as 'black henna', darkens the pigment and precipitates the drying process. PPD is a potent contact allergen associated with a high incidence of hypersensitivity reactions. Despite treatment the patient was left with extensive keloid scarring in the pattern of the henna tattoo.

  4. Severe type IV hypersensitivity to ‘black henna’ tattoo

    Science.gov (United States)

    Vasilakis, Vasileios; Knight, Bernice; Lidder, Satnam; Frankton, Sarah

    2010-01-01

    A 16-year-old Bangladeshi girl presented with a 9-day history of an extensive pruritic, erythematous, papulovesicular skin eruption to both forearms. Appearance was 5 days following application of a home-made henna preparation. Examination revealed ulceration and scabbing along the whole henna pattern and early keloid formation. A diagnosis of type IV delayed hypersensitivity reaction superimposed by infection was initially made. As in this case, home-made henna preparations commonly combine commercial henna with black hair dye, paraphenylenediamine (PPD). PPD, widely known as ‘black henna’, darkens the pigment and precipitates the drying process. PPD is a potent contact allergen associated with a high incidence of hypersensitivity reactions. Despite treatment the patient was left with extensive keloid scarring in the pattern of the henna tattoo. PMID:22778139

  5. Domain and basement membrane specificity of a monoclonal antibody against chicken type IV collagen

    OpenAIRE

    1982-01-01

    A monoclonal antibody, IV-IA8, generated against chicken type IV collagen has been characterized and shown to bind specifically to a conformational-dependent site within a major, triple helical domain of the type IV molecule. Immunohistochemical localization of the antigenic determinant with IV-IA8 revealed that the basement membranes of a variety of chick tissues were stained but that the basement membrane of the corneal epithelium showed little, if any, staining. Thus, basement membranes ma...

  6. Shape and assembly of type IV procollagen obtained from cell culture.

    OpenAIRE

    Oberbäumer, I; Wiedemann, H.; Timpl, R; Kühn, K

    1982-01-01

    Type IV procollagen was isolated from the culture medium of the teratocarcinoma cell line PYS-2 by affinity chromatography on heparin-Sepharose. Immunological studies showed that type IV procollagen is composed of pro-alpha 1(IV) and pro-alpha 2(IV) chains and contains two potential cross-linking sites which are located in the short triple-helical 7S domain and the globular domain NC1 . The 7S domain was also identified as the heparin binding site. Rotary shadowing visualized type IV procolla...

  7. Factors affecting surgical results of Wassel type IV thumb duplications.

    Science.gov (United States)

    Patel, A U C; Tonkin, M A; Smith, B J; Alshehri, A H; Lawson, R D

    2014-11-01

    The aim of this study is to review the outcomes of Wassel type IV thumb duplications with a minimum of one year follow-up, and to identify any factors that may compromise the quality of results. Forty one patients (42 thumbs) returned for assessment of thumb alignment, metacarpophalangeal joint and interphalangeal joint stability and motion; carpometacarpal joint motion; pinch and grip strengths; and thumb size. The subjective assessment considered thumb shape and contour, scarring, nail deformity and examiner and patient/parent satisfaction. The mean age at surgery was 16 months and the mean follow-up time was 79 months. Metacarpophalangeal joint and interphalangeal joint mal-alignment was present in 56% and 38% of cases, respectively. Interphalangeal ulnar collateral ligament laxity was significant. Metacarpophalangeal joint and interphalangeal joint motion was significantly decreased. Pinch and grip strength measurements were normal. A novel and comprehensive assessment scale is introduced, which revealed results of one (2.5%) excellent, 23 (59%) good, 14 (36%) fair and one (2.5%) poor. We consider that greater attention should be directed to the avoidance of mal-alignment and instability to improve these results. © The Author(s) 2013.

  8. Methicillin-resistant Staphylococcus aureus Screening PCR adapted to locally emerging variants-Evaluation of novel SCCmec primers.

    Science.gov (United States)

    Herma, Miriam; Petersdorf, Sabine; Henrich, Birgit

    2017-06-01

    Infections with multi-resistant bacteria, such as Methicillin-resistant Staphylococcus aureus (MRSA), represent a world-wide health-care problem. The original MRSA Screening TaqMan PCR was based on the detection of the SCCmec-orfX-junction as described by the group of Huletsky in 2004. In the recent years, this assay increasingly failed to detect new MRSA variants in swab specimens. In this work, we analyzed the usefulness of 17 additional SCCmec primers to increase PCR sensitivity by testing 290 collected samples with negative PCR results and positive MRSA culture in a retrospective analysis, and 380 samples of the daily routine diagnostics. Sequencing of the PCR products revealed that locally new MRSA variants became detectable by nine of these forward primers. Four primers were solely responsible for the detection of 85.4% (117/123) of the PCR products: F13 (n=76), F11 (n=6), F14 (n=15) and F25 (n=8). These four primers were integrated in the Screening PCR and the novel primer collection was validated by testing 71 MRSA isolates, which covered SCCmec types I to VI, 50 MSSA isolates and 100 swab specimens. The sensitivity of MRSA Screening PCR increased from 93% to 98.6% without affecting the detection of the common MRSA strains. Phylogenetic analysis of the PCR products suggests that the adapted MRSA Screening PCR is able to detect SCCmec types I-X, including CA- and LA-MRSA variants by the SCCmec primers F11 and F25. Copyright © 2017 Elsevier GmbH. All rights reserved.

  9. Bacterial type IV secretion: conjugation systems adapted to deliver effector molecules to host cells

    OpenAIRE

    Christie, Peter J.; Vogel, Joseph P.

    2000-01-01

    Several bacterial pathogens utilize conjugation machines to export effector molecules during infection. Such systems are members of the type IV or ‘adapted conjugation’ secretion family. The prototypical type IV system is the Agrobacterium tumefaciens T-DNA transfer machine, which delivers oncogenic nucleoprotein particles to plant cells. Other pathogens, including Bordetella pertussis, Legionella pneumophila, Brucella spp. and Helicobacter pylori, use type IV machines to export effector prot...

  10. Evolution of conjugation and type IV secretion systems.

    Science.gov (United States)

    Guglielmini, Julien; de la Cruz, Fernando; Rocha, Eduardo P C

    2013-02-01

    Genetic exchange by conjugation is responsible for the spread of resistance, virulence, and social traits among prokaryotes. Recent works unraveled the functioning of the underlying type IV secretion systems (T4SS) and its distribution and recruitment for other biological processes (exaptation), notably pathogenesis. We analyzed the phylogeny of key conjugation proteins to infer the evolutionary history of conjugation and T4SS. We show that single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA) conjugation, while both based on a key AAA(+) ATPase, diverged before the last common ancestor of bacteria. The two key ATPases of ssDNA conjugation are monophyletic, having diverged at an early stage from dsDNA translocases. Our data suggest that ssDNA conjugation arose first in diderm bacteria, possibly Proteobacteria, and then spread to other bacterial phyla, including bacterial monoderms and Archaea. Identifiable T4SS fall within the eight monophyletic groups, determined by both taxonomy and structure of the cell envelope. Transfer to monoderms might have occurred only once, but followed diverse adaptive paths. Remarkably, some Firmicutes developed a new conjugation system based on an atypical relaxase and an ATPase derived from a dsDNA translocase. The observed evolutionary rates and patterns of presence/absence of specific T4SS proteins show that conjugation systems are often and independently exapted for other functions. This work brings a natural basis for the classification of all kinds of conjugative systems, thus tackling a problem that is growing as fast as genomic databases. Our analysis provides the first global picture of the evolution of conjugation and shows how a self-transferrable complex multiprotein system has adapted to different taxa and often been recruited by the host. As conjugation systems became specific to certain clades and cell envelopes, they may have biased the rate and direction of gene transfer by conjugation within prokaryotes.

  11. Structural Characterization of Novel Pseudomonas aeruginosa Type IV Pilins

    Energy Technology Data Exchange (ETDEWEB)

    Nguyen, Y.; Jackson, S; Aidoo, F; Junop, M; Burrows, L

    2010-01-01

    Pseudomonas aeruginosa type IV pili, composed of PilA subunits, are used for attachment and twitching motility on surfaces. P. aeruginosa strains express one of five phylogenetically distinct PilA proteins, four of which are associated with accessory proteins that are involved either in pilin posttranslational modification or in modulation of pilus retraction dynamics. Full understanding of pilin diversity is crucial for the development of a broadly protective pilus-based vaccine. Here, we report the 1.6-{angstrom} X-ray crystal structure of an N-terminally truncated form of the novel PilA from strain Pa110594 (group V), which represents the first non-group II pilin structure solved. Although it maintains the typical T4a pilin fold, with a long N-terminal {alpha}-helix and four-stranded antiparallel {beta}-sheet connected to the C-terminus by a disulfide-bonded loop, the presence of an extra helix in the {alpha}{beta}-loop and a disulfide-bonded loop with helical character gives the structure T4b pilin characteristics. Despite the presence of T4b features, the structure of PilA from strain Pa110594 is most similar to the Neisseria gonorrhoeae pilin and is also predicted to assemble into a fiber similar to the GC pilus, based on our comparative pilus modeling. Interactions between surface-exposed areas of the pilin are suggested to contribute to pilus fiber stability. The non-synonymous sequence changes between group III and V pilins are clustered in the same surface-exposed areas, possibly having an effect on accessory protein interactions. However, based on our high-confidence model of group III PilA{sub PA14}, compensatory changes allow for maintenance of a similar shape.

  12. Expression of 72 kilodalton type IV collagenase (gelatinase A) in benign and malignant ovarian tumors.

    Science.gov (United States)

    Autio-Harmainen, H; Karttunen, T; Hurskainen, T; Höyhtyä, M; Kauppila, A; Tryggvason, K

    1993-09-01

    72 Kilodalton (kd) type IV collagenase is a matrix metalloproteinase that specifically cleaves type IV collagen molecules. The enzyme has been postulated to have an important role in the invasion and spread of malignant tumors. In situ hybridization was used to study the expression of the 72 kd type IV collagenase mRNA in 24 benign, 2 semimalignant, and 15 malignant ovarian tumors and in 5 metastases of ovarian serous adenocarcinomas. The results were correlated with the expression of the mRNA for the alpha 1(IV) chain of type IV collagen and with the corresponding immunohistochemical distribution of the enzyme. The results showed that the more malignant an ovarian tumor was, the more clearly mRNA expressions for both 72 kd type IV collagenase and the alpha 1(IV) chain could be detected in tumor cells. The expression of both types of mRNAs was localized within the cells of tumor stroma and occurred mainly in fibroblasts and vascular endothelial cells. Epithelial tumor cells only rarely expressed these mRNAs. Immunohistochemical stainings localized the 72 kd collagenase as well to the stromal cells as to the epithelial cells of both benign and malignant tumors. The findings indicate that genes for the 72 kd type IV collagenase and for its substrate are simultaneously active in the same cells of the tumor stroma. The difference in the in situ hybridization and immunohistochemical findings could be explained by a possible variation in the metabolic balance between synthesis and accumulation of the protein in different cell types. It can also be proposed that the activity of the 72 kd type IV collagenase would be mediated through a receptor-like mechanism present on epithelial cells which could bind the 72 kd type IV collagenase synthesized elsewhere. There is also a possibility that the gelatinolytic activity of the mesenchymally synthesized 72 kd type IV collagenase would be consumed to degrade extracellular matrix proteins other than basement membranes.

  13. Type IV Neonatal Early-Onset Group B Streptococcal Disease in a United States Hospital▿

    Science.gov (United States)

    Puopolo, Karen M.; Madoff, Lawrence C.

    2007-01-01

    Group B streptococcus (GBS) serotypes causing neonatal disease vary by geographic region. Surveillance at the Brigham and Women's Hospital in Boston, Massachusetts, revealed a case of neonatal early-onset sepsis caused by type IV GBS. Neonatal type IV disease occurs in the Middle East but has not recently been described in U.S. infants. PMID:17267636

  14. Type IV neonatal early-onset group B streptococcal disease in a United States hospital.

    Science.gov (United States)

    Puopolo, Karen M; Madoff, Lawrence C

    2007-04-01

    Group B streptococcus (GBS) serotypes causing neonatal disease vary by geographic region. Surveillance at the Brigham and Women's Hospital in Boston, Massachusetts, revealed a case of neonatal early-onset sepsis caused by type IV GBS. Neonatal type IV disease occurs in the Middle East but has not recently been described in U.S. infants.

  15. Evolutionary Origin of the Staphylococcal Cassette Chromosome mec (SCCmec)

    DEFF Research Database (Denmark)

    Rolo, Joana; Worning, Peder; Nielsen, Jesper Boye

    2017-01-01

    Several lines of evidence indicate that the most primitive staphylococcal species, those of the Staphylococcus sciuri group, were involved in the first stages of evolution of the staphylococcal cassette chromosome mec (SCCmec), the genetic element carrying the β-lactam resistance gene mecA. Howev....... aureus clones) originated in S. sciuri possibly by a recombination event in a human host or a human-created environment and later was transferred to S. aureus....

  16. [Distribution of collagen types III and IV in human placental villi].

    Science.gov (United States)

    Nanaev, A K; Rukosuev, V S; Milovanov, A P; Fokin, E I; Shirinskiĭ, V P

    1989-02-01

    Immunofluorescent examination showed more significant accumulation of interstitial collagen type III in the stroma of mature placenta compared with immature one. Localization of membrane collagen type IV was found neither in basal membranes of epithelium and villous vessels of mature term placenta, nor in their stroma. The described patterns of distribution of collagen types III and IV in human placenta villi were proved by immunoelectronmicroscopic method.

  17. Distribution of Basement Membrane Molecules, Laminin and Collagen Type IV, in Normal and Degenerated Cartilage Tissues

    Science.gov (United States)

    Toh, Wei Seong; Gomoll, Andreas H.; Olsen, Bjørn Reino; Spector, Myron

    2014-01-01

    Objective: The objective of the present study was to investigate the presence and distribution of 2 basement membrane (BM) molecules, laminin and collagen type IV, in healthy and degenerative cartilage tissues. Design: Normal and degenerated tissues were obtained from goats and humans, including articular knee cartilage, the intervertebral disc, and meniscus. Normal tissue was also obtained from patella-tibial enthesis in goats. Immunohistochemical analysis was performed using anti-laminin and anti–collagen type IV antibodies. Human and goat skin were used as positive controls. The percentage of cells displaying the pericellular presence of the protein was graded semiquantitatively. Results: When present, laminin and collagen type IV were exclusively found in the pericellular matrix, and in a discrete layer on the articulating surface of normal articular cartilage. In normal articular (hyaline) cartilage in the human and goat, the proteins were found co-localized pericellularly. In contrast, in human osteoarthritic articular cartilage, collagen type IV but not laminin was found in the pericellular region. Nonpathological fibrocartilaginous tissues from the goat, including the menisci and the enthesis, were also positive for both laminin and collagen type IV pericellularly. In degenerated fibrocartilage, including intervertebral disc, as in degenerated hyaline cartilage only collagen type IV was found pericellularly around chondrocytes but with less intense staining than in non-degenerated tissue. In calcified cartilage, some cells were positive for laminin but not type IV collagen. Conclusions: We report differences in expression of the BM molecules, laminin and collagen type IV, in normal and degenerative cartilaginous tissues from adult humans and goats. In degenerative tissues laminin is depleted from the pericellular matrix before collagen type IV. The findings may inform future studies of the processes underlying cartilage degeneration and the functional

  18. Influence of shelf life on the setting time of type IV gypsum

    Science.gov (United States)

    Hapsari, M. L.; Irawan, B.; Damiyanti, M.

    2017-08-01

    Although expired materials can exhibit a deterioration in their properties, expired type IV gypsum can still be found on the market. In order to evaluate the influence of the shelf life on its setting time, two groups of type IV gypsum (GC Fuji rock EP) with different expiration dates were used in this research. The setting time tests were done in a mold using a Vicat Needle apparatus. The results of the statistical analysis showed a significant difference (p<0.05) between the two different expiration date groups. Therefore, the shelf life did influence the setting time of the type IV gypsum.

  19. Prognostic significance of the expression of MUC1 and collagen type IV in advanced gastric carcinoma.

    Science.gov (United States)

    Ando, H; Aihara, R; Ohno, T; Ogata, K; Mochiki, E; Kuwano, H

    2009-08-01

    Scirrhous gastric carcinoma is characterized by excessive deposition of collagen in the stroma. However, the clinical significance of this fibrosis of the stomach has not been clarified. The aim of this study was to examine the fibrotic mechanism in several histological types of gastric carcinoma, and the combination of MUC1 and collagen type IV as a possible predictor of patient survival. One hundred and two paraffin-embedded specimens of gastric carcinoma were examined by immunohistochemical staining using monoclonal antibodies against collagen type IV and MUC1. Collagen type IV-positive expression was significantly associated with depth of wall penetration (P = 0.025) and stage (P = 0.023). There was a significant relationship between MUC1-positive expression and interstitial collagen type IV-positive expression (P = 0.035). Survival was shorter for patients with the combination of MUC1-positive expression and interstitial collagen type IV-negative expression than for those with other expression patterns. In patients with differentiated-type advanced gastric carcinoma, the combination of MUC1-positive and interstitial collagen type IV-negative expression may be a marker of unfavourable prognosis. Copyright 2009 British Journal of Surgery Society Ltd. Published by John Wiley & Sons, Ltd.

  20. Structural Characterization of Outer Membrane Components of the Type IV Pili System in Pathogenic Neisseria

    NARCIS (Netherlands)

    Jain, Samta; Moscicka, Katarzyna B.; Bos, Martine P.; Pachulec, Emilia; Stuart, Marc C. A.; Keegstra, Wilko; Boekema, Egbert J.; van der Does, Chris; B. Mościcka, K.; Ahmed, Niyaz

    2011-01-01

    Structures of the type IV pili secretin complexes from Neisseria gonorrhoeae and Neisseria meningitidis, embedded in outer membranes were investigated by transmission electron microscopy. Single particle averaging revealed additional domains not observed previously. Secretin complexes of N.

  1. The distribution of collagen types I, III, and IV in normal and malignant colorectal mucosa.

    Science.gov (United States)

    Hilska, M; Collan, Y; Peltonen, J; Gullichsen, R; Paajanen, H; Laato, M

    1998-06-01

    To compare the distribution of interstitial collagens (type I and III) and basement membrane collagen (type IV) in cancerous and normal colon. Retrospective study. University hospital, Finland. 13 patients with colorectal cancer of different stages and grades. Indirect immunofluorescence labelling for type I, III, and IV collagens of fresh frozen tissue samples, both normal and cancerous, cut into serial sections 6 microm thick. In normal mucosa, the epithelial basement membrane showed an intense immunoreaction for type IV collagen. Type I and III collagens were localised to the interstitial stroma underlying it. The membrane in cancer samples was characterised by discontinuities and thinning as estimated by immunolabelling for type IV collagen. Furthermore, immediately adjacent to the membrane type I and III collagen positivity was fragmented. The cancerous stroma showed a strong positive immunosignal for type I and III collagens. Both the epithelial basement membrane and the collagenous matrix immediately beneath it are degraded in malignant tissue. This may suggest the simultaneous activation of several degradative enzymes (as type I and III collagens are at least in part degraded by different enzymes from type IV collagen) or alterations in the expression of collagen subtypes in normal compared with malignant tissue.

  2. An interesting case of type IV hyperlipoproteinemia in a pregnant woman with gestational diabetes mellitus

    OpenAIRE

    S V Shilpa; Prashant Vishwanath; K B Suma; Ujwal B Upadya

    2013-01-01

    We report here a case of asymptomatic type IV hyperlipoproteinemia (HLP) in a 28-year-old pregnant woman with gestational diabetes mellitus (GDM) in the third trimester of pregnancy that was diagnosed incidentally due to the presence of milky white serum. Her total cholesterol and triglyceride values were highly elevated, which led to the provisional diagnosis of hyperlipidemia, and, on further evaluating the patient with lipoprotein electrophoresis, a diagnosis of type IV HLP was made. Her t...

  3. Evaluation of the Various Drying Methods on Surface Hardness of Type IV Dental Stone

    OpenAIRE

    Sudhakar, A.; Srivatsa, G.; Shetty, Rohit; Rajeswari, C L; Manvi, Supriya

    2015-01-01

    Background: Studies regarding the effect of various methods to increase the surface hardness of Type IV dental stone are not conclusive. Therefore, this study was carried out to evaluate the effect of air drying, micro oven drying and die hardener on surface hardness of Type IV dental stone. Materials and Methods: A standard metal die was fabricated; polyvinyl siloxane impression material was used to make the molds of metal die. A total of 120 specimens were obtained from two different die st...

  4. Tacrolimus-Induced Type IV Renal Tubular Acidosis following Liver Transplantation

    Directory of Open Access Journals (Sweden)

    Christopher Schmoyer

    2017-01-01

    Full Text Available Calcineurin inhibitors remain an integral component of immunosuppressive therapy regimens following solid organ transplantation. Although nephrotoxicity associated with these agents is well documented, type IV renal tubular acidosis is a rare and potentially underreported complication following liver transplantation. Hepatologists must be able to recognize this adverse effect as it can lead to fatal hyperkalemia. We describe a case of tacrolimus-induced hyperkalemic type IV renal tubular acidosis in a patient following an orthotopic liver transplant for alcoholic cirrhosis.

  5. Autoantibodies against basement membrane collagen type IV are associated with myocardial infarction

    OpenAIRE

    McLeod, Olga; Dunér, Pontus; Samnegård, Ann; Tornvall, Per; Nilsson, Jan; Hamsten, Anders; Bengtsson, Eva

    2014-01-01

    Background: Collagen type IV is the major constituent of basement membranes underlying endothelial cells and is important for endothelial cell attachment and function. Autoantibodies against native collagen type IV have been found in various autoimmune diseases. Oxidation of LDL in the vascular wall results in the formation of reactive aldehydes, which could modify surrounding matrix proteins. Like oxidized LDL, these modified matrix proteins are likely to induce immune responses. We examined...

  6. Struggling with a Gastric Volvulus Secondary to a Type IV Hiatal Hernia

    Directory of Open Access Journals (Sweden)

    Dafnomilis George

    2010-01-01

    Full Text Available Type IV hiatal hernias are characterized by herniation of the stomach along with associated viscera such as the spleen, colon, small bowel, and pancreas through the esophageal hiatus. They are relatively rare, representing only about 5%–7% of all hernias, and can be associated with severe complications. We report a 71-year-old veteran wrestler who presented to our department with a type IV paraesophageal hernia containing a gastric volvulus and treated successfully with emergency operation.

  7. Increased expression of the type IV secretion system in piliated Neisseria gonorrhoeae variants.

    Science.gov (United States)

    Salgado-Pabón, Wilmara; Du, Ying; Hackett, Kathleen T; Lyons, Katelynn M; Arvidson, Cindy Grove; Dillard, Joseph P

    2010-04-01

    Neisseria gonorrhoeae produces a type IV secretion system that secretes chromosomal DNA. The secreted DNA is active in the transformation of other gonococci in the population and may act to transfer antibiotic resistance genes and variant alleles for surface antigens, as well as other genes. We observed that gonococcal variants that produced type IV pili secreted more DNA than variants that were nonpiliated, suggesting that the process may be regulated. Using microarray analysis, we found that a piliated strain showed increased expression of the gene for the putative type IV secretion coupling protein TraD, whereas a nonpiliated variant showed increased expression of genes for transcriptional and translational machinery, consistent with its higher growth rate compared to that of the piliated strain. These results suggested that type IV secretion might be controlled by either traD expression or growth rate. A mutant with a deletion in traD was found to be deficient in DNA secretion. Further mutation and complementation analysis indicated that traD is transcriptionally and translationally coupled to traI, which encodes the type IV secretion relaxase. We were able to increase DNA secretion in a nonpiliated strain by inserting a gene cassette with a strong promoter to drive the expression of the putative operon containing traI and traD. Together, these data suggest a model in which the type IV secretion system apparatus is made constitutively, while its activity is controlled through regulation of traD and traI.

  8. Increased Expression of the Type IV Secretion System in Piliated Neisseria gonorrhoeae Variants▿

    Science.gov (United States)

    Salgado-Pabón, Wilmara; Du, Ying; Hackett, Kathleen T.; Lyons, Katelynn M.; Arvidson, Cindy Grove; Dillard, Joseph P.

    2010-01-01

    Neisseria gonorrhoeae produces a type IV secretion system that secretes chromosomal DNA. The secreted DNA is active in the transformation of other gonococci in the population and may act to transfer antibiotic resistance genes and variant alleles for surface antigens, as well as other genes. We observed that gonococcal variants that produced type IV pili secreted more DNA than variants that were nonpiliated, suggesting that the process may be regulated. Using microarray analysis, we found that a piliated strain showed increased expression of the gene for the putative type IV secretion coupling protein TraD, whereas a nonpiliated variant showed increased expression of genes for transcriptional and translational machinery, consistent with its higher growth rate compared to that of the piliated strain. These results suggested that type IV secretion might be controlled by either traD expression or growth rate. A mutant with a deletion in traD was found to be deficient in DNA secretion. Further mutation and complementation analysis indicated that traD is transcriptionally and translationally coupled to traI, which encodes the type IV secretion relaxase. We were able to increase DNA secretion in a nonpiliated strain by inserting a gene cassette with a strong promoter to drive the expression of the putative operon containing traI and traD. Together, these data suggest a model in which the type IV secretion system apparatus is made constitutively, while its activity is controlled through regulation of traD and traI. PMID:20139191

  9. Laparoscopic treatment of type III and IV hiatal hernia – authors’ experience

    Science.gov (United States)

    Grzesiak-Kuik, Agata; Pędziwiatr, Michał; Budzyński, Andrzej

    2014-01-01

    Introduction There are four types of hiatal hernias, and diagnosis is established on the basis of gastroscopy in the majority of cases. Type III represents a mixed type in which the abdominal esophagus as well as the gastric cardia and fundus protrude into the thorax through the pathologically widened esophageal hiatus. Type IV, the so-called upside down stomach, can be considered an evolutionary form of type III, and refers to herniation of nearly the whole stomach (except for the cardia and pylorus) into the thorax. Types III and IV of hiatal hernias represent a group of rare diaphragmatic defects; thus, most centers do not possess considerable experience in their treatment. Frequently, laparoscopic treatment is implemented, although, according to some authors, conversion to laparotomy, thoracotomy, or thoracolaparotomy is necessary in selected cases. Aim To analyze the outcomes of laparoscopic treatment of the largest hiatal hernias, i.e. type III and IV hernias. Material and methods A total of 25 patients diagnosed with type III and IV hiatal hernia were included in further analysis. Results As many as 19 out of 25 patients (76%) assessed the outcome of the surgery as evidently positive and reported marked improvement in the quality of life. Conclusions The laparoscopic technique constitutes an excellent and safe method of repair of even the most complex defects in the esophageal hiatus. Therefore, the minimally invasive technique combined with an anti-reflux procedure should be the method of choice in patients with type III and IV hernia. PMID:25097681

  10. Distribution of types I, II, III, IV and V collagen in normal and keratoconus corneas.

    Science.gov (United States)

    Nakayasu, K; Tanaka, M; Konomi, H; Hayashi, T

    1986-01-01

    By using type-specific antibodies to types I, II, III, IV and V collagens, distribution of distinct types of collagen in normal human cornea as well as keratoconus cornea were examined by indirect immunofluorescence microscopy. In normal human cornea, immunohistochemical evidence supported the previous biochemical finding that type I collagen was the major type of collagen in human corneal stroma. No reaction was observed to anti-type II collagen antibody in the whole cornea. Anti-type III collagen antibody reacted with the corneal stroma in a similar fashion as that of anti-type I collagen antibody. Type IV collagen was observed in the basement membrane of the corneal epithelium and in Descemet's membrane. Anti-type V collagen antibody also reacted with the corneal stroma diffusely. Bowman's membrane was strongly stained only with he anti-type V collagen antibody. For further details of the distribution of type I, type III and V collagens in human corneal stroma, immunoelectron microscopic study was undertaken. The positive reaction products of anti-type I and anti-type III collagen antibodies were located on the collagen fibrils, while that of anti-type V collagen antibody was either on or close to collagen fibrils. In keratoconus cornea, no difference was observed in terms of the distribution of type I, III and V collagens, while the disruptive and excrescent distribution of type IV collagen was noted in the basement membrane of the corneal epithelium.

  11. Cloning, expression, and activity of type IV antifreeze protein from cultured subtropical olive flounder (Paralichthys olivaceus

    Directory of Open Access Journals (Sweden)

    Jong Kyu Lee

    2016-10-01

    Full Text Available Abstract Antifreeze proteins (AFPs lower the freezing point but not the melting point of aqueous solutions by inhibiting the growth of ice crystals via an adsorption-inhibition mechanism. However, the function of type IV AFP (AFP IV is questionable, as its antifreeze activity is on the verge of detectable limits, its physiological concentration in adult fish blood is too low to function as a biological antifreeze, and its homologues are present even in fish from tropic oceans as well as freshwater. Therefore, we speculated that AFP IV may have gained antifreeze activity not by selective pressure but by chance. To test this hypothesis, we cloned, expressed, and assayed AFP IV from cultured subtropical olive flounder (Paralichthys olivaceus, which do not require antifreeze protein for survival. Among the identified expressed sequence tags of the flounder liver sample, a 5′-deleted complementary DNA (cDNA sequence similar to the afp4 gene of the longhorn sculpin was identified, and its full-length cDNA and genome structure were examined. The deduced amino acid sequence of flounder AFP IV shared 55, 53, 52, and 49 % identity with those of Pleuragramma antarcticum, Myoxocephalus octodecemspinosus, Myoxocephalus scorpius, and Notothenia coriiceps, respectively. Furthermore, the genomic structure of this gene was conserved with those of other known AFP IVs. Notably, the recombinant AFP IV showed a weak but distinct thermal hysteresis of 0.07 ± 0.01 °C at the concentration of 0.5 mg/mL, and ice crystals in an AFP IV solution grew star-shaped, which are very similar to those obtained from other polar AFP IVs. Taken together, our results do not support the hypothesis of evolution of AFP IV by selective pressure, suggesting that the antifreeze activity of AFP IV may have been gained by chance.

  12. Type IV Ehlers-Danlos Syndrome: A Surgical Emergency? A Case of Massive Retroperitoneal Hemorrhage

    Science.gov (United States)

    Chun, Stephen G; Pedro, Patrick; Yu, Mihae; Takanishi, Danny M

    2011-01-01

    Retroperitoneal hemorrhagic bleeding is a known manifestation of Type-IV Ehlers-Danlos Syndrome that is caused by loss-of-function mutations of the pro-alpha-1 chains of type III pro-collagen (COL3A1) resulting in vascular fragility. A number of previous reports describe futile surgical intervention for retroperitoneal bleeding in Type-IV Ehlers-Danlos Syndrome with high post-operative mortality, although the rarity of retroperitoneal bleeding associated with Type-IV Ehlers-Danlos Syndrome precludes an evidence-based approach to clinical management. We report a 23-year-old male with history of Type-IV Ehlers-Danlos Syndrome who presented with severe abdominal pain and tachycardia following an episode of vomiting. Further work-up of his abdominal pain revealed massive retroperitoneal bleeding by CT-scan of the abdomen. Given numerous cases of catastrophic injury caused by surgical intervention in Type-IV Ehlers-Danlos Syndrome, the patient was treated non-operatively, and the patient made a full recovery. This case suggests that even in cases of large retroperitoneal hemorrhages associated with Ehlers-Danlos Syndrome, it may not truly represent a surgical emergency. PMID:21966332

  13. Cost-Effectiveness and Efficacy of spa, SCCmec, and PVL Genotyping of Methicillin-Resistant Staphylococcus aureus as Compared to Pulsed-Field Gel Electrophoresis

    Science.gov (United States)

    Li, Vincent; Chui, Linda; Louie, Lisa; Simor, Andrew; Golding, George R.; Louie, Marie

    2013-01-01

    Pulsed-field gel electrophoresis (PFGE) is a valuable molecular typing assay used for methicillin-resistant Staphylococcus aureus (MRSA) surveillance and genotyping. However, there are several limitations associated with PFGE. In Alberta, Canada, the significant increase in the number of MRSA isolates submitted to the Provincial Laboratory for Public Health (ProvLab) for PFGE typing led to the need for an alternative genotyping method. In this study, we describe the transition from PFGE to Staphylococcus protein A (spa), Staphylococcal cassette chromosome (SCCmec), and Panton-Valentine leukocidin (PVL) typing. A total of 1915 clinical MRSA isolates collected from 2005 to 2009 were used to develop and validate an algorithm for assigning PFGE epidemic types using spa, SCCmec, and PVL typing and the resulting data was used to populate a new Alberta MRSA typing database. An additional 12620 clinical MRSA isolates collected from 2010 to 2012 as part of ongoing routine molecular testing at ProvLab were characterized using the new typing algorithm and the Alberta MRSA typing database. Switching to spa, SCCmec, and PVL from PFGE typing substantially reduced hands-on and turn-around times while maintaining historical PFGE epidemic type designations. This led to an approximate $77,000 reduction in costs from 2010 to 2012. PFGE typing is still required for a small subset of MRSA isolates that have spa types that are rare, novel, or associated with more than one PFGE epidemic type. PMID:24244440

  14. Dual (type IV) left anterior descending artery | Baskan | SA Journal of ...

    African Journals Online (AJOL)

    Congenital coronary artery anomalies are uncommon. Dual left anterior descending coronary artery (LAD) is defined as the presence of two LADs within the anterior interventricular sulcus (AIVS), and is classified into four types. Type IV is a rarely reported subtype and differs from the others, with a long LAD originating from ...

  15. Abnormal distribution of collagen type IV in extrahepatic bile duct carcinoma.

    Science.gov (United States)

    Chen, Y; Sasatomi, E; Satoh, T; Miyazaki, K; Tokunaga, O

    2000-11-01

    The present study investigated the pathogenesis of desmoplastic stroma formation, which is characteristic of most bile duct carcinomas and other scirrhous carcinomas. Using immunohistochemical analysis, the expression of collagen types I and IV, laminin and TGF-beta1 was examined in human extrahepatic bile duct carcinoma and compared with gastric and colon carcinoma. In addition to delineating the basement membranes of carcinoma nests and blood vessels, collagen type IV was present along the thick bundles of collagenous fibers in the stroma of extrahepatic bile duct carcinoma and scirrhous gastric carcinoma. The immunoreactivity of collagen type IV was strong in the adjacent or surrounding interstitium of tumor cell nests, but was absent or weak in older, more central portions of the tumor that contained sclerotic collagen. In situ hybridization demonstrated active expression of collagen alpha1(IV) mRNA in extrahepatic bile duct carcinoma and scirrhous gastric carcinoma cells. These results suggest that, although collagen type IV is typically a component of the basement membrane, it is expressed in the interstitial stroma of extrahepatic bile duct carcinoma and scirrhous gastric carcinoma where it may play a role in desmoplastic stroma formation.

  16. The Role of Type IV Collagen in Developing Lens in Mouse Fetuses

    Directory of Open Access Journals (Sweden)

    Mehdi Jalali

    2009-09-01

    Full Text Available Objective(sExtracellular matrix (ECM and basement membrane (BM play important roles in many developmental processes during development and after birth. Among the components of the BM, collagen fibers specially type IV are the most important parts. The aim of this study was to determine the time when collagen type IV appears in the BM of lens structure during mouse embryonic development.Materials and MethodsIn this experimental study, 22 female Balb/C mice were randomly selected and were kept under normal condition, finding vaginal plug was assumed as day zero of pregnancy. From embryonic day 10 to 20, all specimens were sacrificed by cervical dislocation and their heads were fixed, serially sectioned and immunohistochemistry study for tracing collagen type IV in lens were carried out.ResultsOur data revealed that collagen type IV appeared at the early stage of gestation day 12 in BM of anterior epithelial lens cells and the amount of this protein gradually increased until days 15-17 in ECM and posterior capsule epithelium. After this period, severe reaction was not observed in any part of the lens.ConclusionThese findings establish the important role of collagen IV in developing optic cup and any changes during critical period of pregnancy may be result in severe visual system defect

  17. Spontaneous Carotid-Cavernous Fistula in the Type IV Ehlers-Danlos Syndrome

    Science.gov (United States)

    Kim, Jeong Gyun; Cho, Won-Sang; Kim, Jeong Eun

    2014-01-01

    Ehlers-Danlos syndrome (EDS) is a rare inherited connective disease. Among several subgroups, type IV EDS is frequently associated with spontaneous catastrophic bleeding from a vascular fragility. We report on a case of carotid-cavernous fistula (CCF) in a patient with type IV EDS. A 46-year-old female presented with an ophthalmoplegia and chemosis in the right eye. Subsequently, seizure and cerebral infarction with micro-bleeds occurred. CCF was completely occluded with transvenous coil embolization without complications. Thereafter, the patient was completely recovered. Transvenous coil embolization can be a good treatment of choice for spontaneous CCF with type IV EDS. However, every caution should be kept during invasive procedure. PMID:24653803

  18. Clinical application of antenatal genetic diagnosis of osteogenesis imperfecta type IV.

    Science.gov (United States)

    Yuan, Jing; Li, Song; Xu, YeYe; Cong, Lin

    2015-04-02

    Clinical analysis and genetic testing of a family with osteogenesis imperfecta type IV were conducted, aiming to discuss antenatal genetic diagnosis of osteogenesis imperfecta type IV. Preliminary genotyping was performed based on clinical characteristics of the family members and then high-throughput sequencing was applied to rapidly and accurately detect the changes in candidate genes. Genetic testing of the III5 fetus and other family members revealed missense mutation in c.2746G>A, pGly916Arg in COL1A2 gene coding region and missense and synonymous mutation in COL1A1 gene coding region. Application of antenatal genetic diagnosis provides fast and accurate genetic counseling and eugenics suggestions for patients with osteogenesis imperfecta type IV and their families.

  19. Oxyhydroxy Silicate Colloids: A New Type of Waterborne Actinide(IV) Colloids

    Science.gov (United States)

    Weiss, Stephan; Hennig, Christoph; Brendler, Vinzenz; Ikeda‐Ohno, Atsushi

    2016-01-01

    Abstract At the near‐neutral and reducing aquatic conditions expected in undisturbed ore deposits or in closed nuclear waste repositories, the actinides Th, U, Np, and Pu are primarily tetravalent. These tetravalent actinides (AnIV) are sparingly soluble in aquatic systems and, hence, are often assumed to be immobile. However, AnIV could become mobile if they occur as colloids. This review focuses on a new type of AnIV colloids, oxyhydroxy silicate colloids. We herein discuss the chemical characteristics of these colloids and the potential implication for their environmental behavior. The binary oxyhydroxy silicate colloids of AnIV could be potentially more mobile as a waterborne species than the well‐known mono‐component oxyhydroxide colloids. PMID:27957406

  20. Epidemiology of hereditary sensory and autonomic neuropathy type IV and V in Japan.

    Science.gov (United States)

    Haga, Nobuhiko; Kubota, Masaya; Miwa, Zenzo

    2013-04-01

    Hereditary sensory and autonomic neuropathy (HASN) refers to a group of rare congenital disorders characterized by loss of pain sensation and other sensory or autonomic abnormalities. Among them, a relatively large proportion of patients with HSAN type IV, which is accompanied by anhidrosis and intellectual disability, are reported from Israel and Japan. HSAN type V, with normal sweating and mental development, is rarely reported in Japan. In 2009, we founded a research group for congenital insensitivity to pain and performed the first epidemiological survey of HSAN types IV and V in Japan. Questionnaires were sent to a total of 3,488 certified training institutions of five nationwide medical societies comprising pediatricians, neurologists, orthopedic surgeons, and dentists. Answers were obtained from 1,610 institutions, and 192 HSAN patients (152 with type IV and 28 with type V) were reported from 105 institutions. After excluding duplicated patients, we identified a total of 62 current, 36 past, and five deceased patients for HSAN-IV, and a total of 14 current, 13 past, and 0 deceased patients for HSAN-V. Using these figures, we estimated that the number of Japanese patients with HSAN types IV and V as 130-210 and 30-60 patients, respectively. We identified no gender differences, and patients with a family history of the disorder were limited to affected siblings in both conditions. Most patients with HSAN-IV were 5-40 years of age, whereas half of the patients with HSAN-V were 40 years or older. Copyright © 2013 Wiley Periodicals, Inc.

  1. Cardiac arrest after anesthetic management in a patient with hereditary sensory autonomic neuropathy type IV

    Directory of Open Access Journals (Sweden)

    Ergül Yakup

    2011-01-01

    Full Text Available Hereditary sensory autonomic neuropathy type IV is a rare disorder with an autosomal recessive transmission and characterized by self-mutilation due to a lack in pain and heat sensation. Recurrent hyperpyrexia and anhydrosis are seen in patients as a result of a lack of sweat gland innervation. Self-mutilation and insensitivity to pain result in orthopedic complications and patients undergone recurrent surgical interventions with anesthesia. However, these patients are prone to perioperative complications such as hyperthermia, hypothermia, and cardiac complications like bradycardia and hypotension. We report a 5-year-old boy with hereditary sensory autonomic neuropathy type IV, developing hyperpyrexia and cardiac arrest after anesthesia.

  2. Dipeptidyl peptidase IV inhibitors: a promising new therapeutic approach for the management of type 2 diabetes

    DEFF Research Database (Denmark)

    Deacon, Carolyn F; Holst, Jens J

    2005-01-01

    of appetite. Glucagon-like peptide-1 is, however, extremely rapidly inactivated by the serine peptidase, dipeptidyl peptidase IV, so that the native peptide is not useful clinically. A new approach to utilise the beneficial effects of glucagon-like peptide-1 in the treatment of type 2 diabetes has been...... the development of orally active dipeptidyl peptidase IV inhibitors. Preclinical studies have demonstrated that this approach is effective in enhancing endogenous levels of glucagon-like peptide-1, resulting in improved glucose tolerance in glucose-intolerant and diabetic animal models. In recent studies of 3......-12 months duration in patients with type 2 diabetes, dipeptidyl peptidase IV inhibitors have proved efficacious, both as monotherapy and when given in combination with metformin. Fasting and postprandial glucose concentrations were reduced, leading to reductions in glycosylated haemoglobin levels, while...

  3. Stromal assemblies containing collagen types IV and VI and fibronectin in the developing embryonic avian cornea.

    Science.gov (United States)

    Fitch, J M; Birk, D E; Linsenmayer, C; Linsenmayer, T F

    1991-04-01

    The morphogenesis of type IV collagen-containing structures in the stromal matrix of the developing avian cornea was investigated using immunofluorescence and immunoelectron microscopic histochemistry. Two forms of type IV collagen-containing structures were seen; these differed in their probable origin, structure, molecular composition, and developmental fate. The major form of stromal type IV collagen-containing material, termed "strings," was observed only after swelling of the primary stroma and the onset of mesenchymal invasion. These strings are presumed to be products of the stromal cells. In immunofluorescence histochemistry they appeared as linear segments of type IV collagen-specific immunoreactivity. In immunoelectron microscopy, they appeared initially as electron-dense sausages of variable length and orientation. They frequently were associated with cell surfaces and, in fortuitous sections, appeared to connect adjacent cells. The strings also contained type VI collagen and fibronectin, but very little, if any, of the basement membrane components laminin and heparin sulfate proteoglycan (HSPG). As the stroma continued to expand in thickness, more of these structures were observed in a radial orientation, becoming quite long and less tortuous. Later in development, as stromal condensation proceeded, they disappeared. We suggest that the strings function to stabilize the stromal matrix, and perhaps to limit the rate and/or extent of stromal expansion, during a phase of rapid swelling and matrix deposition. The other form of type IV collagen-containing stromal material appeared as irregularly shaped plaques of basement membrane-like material identical to those previously described in mature corneas. These are likely derived from the corneal endothelial cells. They contained other basement membrane-associated components (laminin, HSPG) and fibronectin, but not type VI collagen. This material persists in mature corneas as sparse irregular stromal plaques

  4. Sensing of Bacterial Type IV Secretion via the Unfolded Protein Response

    NARCIS (Netherlands)

    de Jong, Maarten F.; Starr, Tregei; Winter, Maria G.; den Hartigh, Andreas B.; Child, Robert; Knodler, Leigh A.; van Dijl, Jan Maarten; Celli, Jean; Tsolis, Renee M.

    2013-01-01

    Host cytokine responses to Brucella abortus infection are elicited predominantly by the deployment of a type IV secretion system (T4SS). However, the mechanism by which the T4SS elicits inflammation remains unknown. Here we show that translocation of the T4SS substrate VceC into host cells induces

  5. Dipeptidyl-peptidase-IV inhibition augments postprandial lipid mobilization and oxidation in type 2 diabetic patients.

    NARCIS (Netherlands)

    Boschmann, M.; Engeli, S.; Dobberstein, K.; Budziarek, P.; Strauss, A.; Boehnke, J.; Sweep, F.C.; Luft, F.C.; He, Y.; Foley, J.E.; Jordan, J.

    2009-01-01

    CONTEXT: Dipeptidyl-peptidase-IV (DPP-4) inhibition increases endogenous GLP-1 activity, resulting in improved glycemic control in patients with type 2 diabetes mellitus. The metabolic response may be explained in part by extrapancreatic mechanisms. OBJECTIVE: We tested the hypothesis that DPP-4

  6. Tibial lengthening for unilateral Crowe type-IV developmental dysplasia of the hip

    Directory of Open Access Journals (Sweden)

    Jun Wan

    2014-01-01

    Conclusions: Tibial lengthening may effectively correct gait and satisfactorily improve body image in young patients with unilateral Crowe type-IV DDH. Mono-lateral external fixator allows for accelerated postoperative rehabilitation and optimal preservation of ankle movements. Lengthening along with intramedullary nails may significantly reduce the external fixation time and the risk of fixator-related complications.

  7. Liver transplantation for glycogen storage disease types I, III, and IV

    NARCIS (Netherlands)

    Matern, D; Starzl, TE; Arnaout, W; Barnard, J; Bynon, JS; Dhawan, A; Emond, J; Haagsma, EB; Hug, G; Lachaux, A; Smit, GPA; Chen, YT

    1999-01-01

    Glycogen storage disease (GSD) types I, III, and IV can be associated with severe liver disease. The possible development of hepatocellular carcinoma and/or hepatic failure make these GSDs potential candidates for liver transplantation. Early diagnosis and initiation of effective dietary therapy

  8. Host Cell Contact-Induced Transcription of the Type IV Fimbria Gene Cluster of Actinobacillus pleuropneumoniae

    NARCIS (Netherlands)

    Boekema, B.K.H.L.; Putten, J.P.M.; Stockhofe-Zurwieden, N.; Smith, H.E.

    2004-01-01

    Type IV pili (Tfp) of gram-negative species share many characteristics, including a common architecture and conserved biogenesis pathway. Much less is known about the regulation of Tfp expression in response to changing environmental conditions. We investigated the diversity of Tfp regulatory

  9. Elective reconstruction of thoracoabdominal aortic aneurysm type IV by transabdominal approach

    Directory of Open Access Journals (Sweden)

    Marjanović Ivan

    2012-01-01

    Full Text Available Introduction. Thoracoabdominal aortic aneurysm (TAAA type IV represents an aortic dilatation from the level of the diaphragmatic hiatus to the iliac arteries branches, including visceral branches of the aorta. In the traditional procedure of TAAA type IV repair, the body is opened using thoractomy and laparotomy in order to provide adequate exposure of the descending thoracic and abdominal aorta for safe aortic reconstruction. Case report. We reported a 71-yearold man with elective reconstruction of the TAAA type IV performed by transabdominal approach. Computed tomography scans angiography revealed a TAAA type IV with diameter of 62 mm in the region of celiac trunk and superior mesenteric artery branching, and the largest diameter of 75 mm in the infrarenal aortic level. The patient comorbidity included a chronic obstructive pulmonary disease and hypertension, therefore he was treated for a prolonged period. In preparation for the planned aortic reconstruction asymptomatic carotid disease (occlusion of the left internal carotid artery and subtotal stenosis of the right internal carotid artery was diagnosed. Within the same intervention percutaneous transluminal angioplasty with stent placement in right internal carotid artery was made. In general, under endotracheal anesthesia and epidural analgesia, with transabdominal approach performed aortic reconstruction with tubular dakron graft 24 mm were, and reimplantation of visceral aortic branches into the graft performed. Postoperative course was uneventful, and the patient was discharged on the postoperative day 17. Control computed tomography scan angiography performed three months after the operation showed vascular state of the patient to be in order. Conclusion. Complete transabdominal approach to TAAA type IV represents an appropriate substitute for thoracoabdominal approach, without compromising safety of the patient. This approach is less traumatic, especially in patients with impaired

  10. Peripheral arthropathy in hereditary sensory and autonomic neuropathy types III and IV.

    Science.gov (United States)

    Feldman, David S; Ruchelsman, David E; Spencer, Daniel B; Straight, Joseph J; Schweitzer, Mark E; Axelrod, Felicia B

    2009-01-01

    To determine the features of the underlying destructive arthropathy in the peripheral joints of children with hereditary sensory and autonomic neuropathy (HSAN) type III and to compare and contrast this to the arthropathy noted in HSAN type IV, as both groups experience decreased pain perception. From a database of 547 patients with HSAN type III and 32 patients with HSAN type IV, we performed a retrospective chart review and radiographic analysis of all patients who presented with joint swelling and deformity. Underlying joint pathology was classified as either osteonecrosis or Charcot arthropathy. In the HSAN type III population, 44 (8%; 22 males and 22 females) of the 547 patients had clinical evidence of arthropathy. In 42 patients, 48 joints demonstrated radiographic evidence of osteonecrosis; 45 (94%) of the 48 joints with osteonecrosis occurred in the lower extremity. In each case of osteonecrosis of the knee (n = 19), isolated involvement of the lateral distal femoral condyle was seen consisting of varying sizes of posterolateral osteochondral fragmentation. In the 32 patients comprising the HSAN type IV population, 18 (56%) were found to have radiographic findings consistent with Charcot arthropathy in a total of 30 affected joints. One patient demonstrated Charcot arthropathy of the spine and subsequent progressive spondylolisthesis. Nine patients (12 joints) also demonstrated osteomyelitis. In patients with HSAN type III, osteonecrosis is the initial lesion preceding destructive arthropathy. Osteonecrosis and osteochondral fragmentation were always isolated at the lateral distal femoral condyle in the knee. This pathology may be amenable to surgical reconstruction and fixation to stabilize the knee and prevent further degeneration. Hereditary sensory and autonomic neuropathy type IV was most commonly associated with Charcot arthropathy or joint subluxation and dislocation. Late secondary changes at the articular surface may make radiographic distinction

  11. Type-IV Antifreeze Proteins are Essential for Epiboly and Convergence in Gastrulation of Zebrafish Embryos

    OpenAIRE

    Xiao, Qing; Xia, Jian-Hong; Zhang, Xiao-Juan; Li, Zhi; Wang, Yang; Zhou, Li; Gui, Jian-Fang

    2014-01-01

    Many organisms in extremely cold environments such as the Antarctic Pole have evolved antifreeze molecules to prevent ice formation. There are four types of antifreeze proteins (AFPs). Type-IV antifreeze proteins (AFP4s) are present also in certain temperate and even tropical fish, which has raised a question as to whether these AFP4s have important functions in addition to antifreeze activity. Here we report the identification and functional analyses of AFP4s in cyprinid fish. Two genes, nam...

  12. Salter-Harris Type III and Type IV Combined Fracture of the Distal Femoral Epiphysis: A Case Report

    Directory of Open Access Journals (Sweden)

    Ali Aydin

    2012-01-01

    Full Text Available Distal femoral physeal fractures are not common but have a high rate of complications. They generally follow one of the patterns described in the Salter-Harris classification. We present a case of combination of Salter-Harris type III and type IV injury. Our case was a 15-year-old boy who had a motor vehicle accident. There was swelling, ecchymosis, severe pain, and valgus deformity, because of medial proximal fracture fragment, on the left knee. We deemed that Salter-Harris type III and type IV combination fracture in our case has not been previously reported. We prepared this paper in consideration of its contribution to the literature.

  13. Influence of temprature moisture and time on dimensional change of stone type IV

    Directory of Open Access Journals (Sweden)

    Sahebi.SM.

    2004-07-01

    Full Text Available Statement of Problem: Different temperature and moisture around Iran and wide usage of stone type IV in dental laboratories in our country were the reasons for us for doing this study. Purpose: In this survey the effect of temperature, moisture and time on dimensional change of dental stone type IV (Velmix were investigated. Materials and Methods: In this experimental study cube shape Metal models (10×10×10mm was made for Taking impression. We used special tray, which has ten holes (15×20×25mm and made impression with condensational silicone in two-step procedure, and poured it with stone type IV. stone cubes were divided to different groups. 3 groups holed in fix moisture and variable tempraure and 5 groups in fix temperature and variable moisture. After 2 hours, 24 hours and one-week dimension of stone cubes were measured. Then analysis was done with ANOVA and dauncan. Results: The following conclusions were achieved: 1-Time has no effect on dimensional changes. 2-Tempreture and moisture has some effect on dimensional changes in stone IV. Increasing of temperature result in stone contraction and increasing in moisture result in stone expansion. Conclusion: The best temperature for least dimensional change is 20C and the best moisture for a least dimensional change is 30 %.

  14. Type IV kerogens as analogues for organic macromolecular materials in aqueously altered carbonaceous chondrites.

    Science.gov (United States)

    Matthewman, Richard; Martins, Zita; Sephton, Mark A

    2013-04-01

    Understanding the processes involved in the evolution of organic matter in the early Solar System requires extensive experimental work. The scientifically valuable carbonaceous chondrites are principal targets for organic analyses, but these meteorites are rare. Meteoritic analog materials available in larger quantities, on which experiments can be performed, would be highly beneficial. The bulk of the organic inventory of carbonaceous chondrites is made up of solvent-insoluble macromolecular material. This high-molecular-weight entity provides a record of thermal and aqueous parent-body alteration of precursor organic structures present at the birth of the Solar System. To identify an effective analogue for this macromolecular material, we analyzed a series of terrestrial kerogens by pyrolysis-gas chromatography-mass spectrometry. Type I and II kerogens are unsuitable analogues owing to their highly aliphatic nature. Type III kerogens show some similarities to meteoritic macromolecular materials but display a substantial biological heritage. Type IV kerogens, in this study derived from Mesozoic paleosols and produced by the reworking and oxidation of organic matter, represent an effective analogue. Some isomeric differences exist between meteoritic macromolecular materials and type IV kerogens, and stepped pyrolysis indicates variations in thermal stability. In addition to being a suitable material for novel experimentation, type IV kerogens also have the potential to aid in the optimization of instruments for deployment on Mars.

  15. Leverage application on Gartland type IV supracondylar humeral fracture in children.

    Science.gov (United States)

    Pei, Xinhong; Mo, Yueqiang; Huang, Peng

    2016-11-01

    Although the most complex management of Gartland type IV supracondylar humeral fracture (SCHF) due to instability, the gold standard of initial treatment remains closed reduction and percutaneous pinning. However, open reduction was inevitable in most published studies. This study reports the outcome of treatment by leverage-assisted closed reduction. Twenty-seven patients were diagnosed as Gartland type IV SCHF during surgery in 214 preoperative Gartland type III fractures. Leverage-assisted reduction with percutaneous lateral pinning was done in these patients after failure of close reduction. Evaluations were performed with radiographic examination, clinical assessment and Flynn's criteria by interview and physical examination. All 27 patients obtained acceptable reduction by leverage-assisted close reduction and percutaneous pinning. The average follow-up was 23.6 (18-30 months). There was no neurovascular complication, infection, nonunion, myositis ossificans or Volkmann's contracture. Evaluation of Baumann's angle was towards varus (74-74.2°). There was no significant difference (p = 0.1876). Flynn's criteria were excellent in 22 (81.5 %) patients, good in four (14.8 %) and fair in one (3.7 %). The rate of excellent and good outcome was 96.3 % and of satisfactory 100 %. We recommend leverage-assisted closed reduction as an option before open reduction in type IV SCHF, not only for gold standard management but also because of satisfactory outcomes and the low incidence of major complications.

  16. Effect of shelf life on compressive strength of type iv gypsum

    Science.gov (United States)

    Kusumastuti, K. S.; Irawan, B.; Damiyanti, M.

    2017-08-01

    Type IV gypsum, as a dental material for an indirect restoration’s working model, should have strength and abrasive-resistant properties. These properties depend on the product’s shelf life and its proper storage, which sometimes are easily missed by sellers. The aim of this research was to observe the effect of shelf life on the compressive strength of type IV gypsum with different production dates. Twenty cylindrical specimens were separated into two groups with different production dates and tested with a universal testing with the crosshead speed of 1 mm per minute and a load of 2,500 kgf. The data were analyzed with independent t-tests. There was a significant difference (p<0.05) in the compressive strength between the two groups with an increase in compressive strength seen in the gypsum that was stored longer.

  17. Type IV traffic ATPase TrwD as molecular target to inhibit bacterial conjugation

    OpenAIRE

    Ripoll-Rozada, Jorge; Garc��a-Cazorla, Yolanda; Getino, Mar��a; Mach��n, Cristina; Sanabria-R��os, David; de la Cruz, Fernando; Cabez��n, Elena; Arechaga, Ignacio

    2016-01-01

    Bacterial conjugation is the main mechanism responsible for the dissemination of antibiotic resistance genes. Hence, the search for specific conjugation inhibitors is paramount in the fight against the spread of these genes. In this pursuit, unsaturated fatty acids have been found to specifically inhibit bacterial conjugation. Despite the growing interest on these compounds, their mode of action and their specific target remain unknown. Here, we identified TrwD, a Type IV secretion traffic AT...

  18. Irradiation Alters MMP-2/TIMP-2 System and Collagen Type IV Degradation in Brain

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Won Hee [School of Biomedical Engineering and Sciences, Virginia Polytechnic Institute and State University, Blacksburg, Virginia (United States); Warrington, Junie P.; Sonntag, William E. [Reynolds Oklahoma Center on Aging, Department of Geriatric Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma (United States); Lee, Yong Woo, E-mail: ywlee@vt.edu [School of Biomedical Engineering and Sciences, Virginia Polytechnic Institute and State University, Blacksburg, Virginia (United States); Department of Biomedical Sciences and Pathobiology, Virginia Polytechnic Institute and State University, Blacksburg, Virginia (United States)

    2012-04-01

    Purpose: Blood-brain barrier (BBB) disruption is one of the major consequences of radiation-induced normal tissue injury in the central nervous system. We examined the effects of whole-brain irradiation on matrix metalloproteinases (MMPs)/tissue inhibitors of metalloproteinases (TIMPs) and extracellular matrix (ECM) degradation in the brain. Methods and Materials: Animals received either whole-brain irradiation (a single dose of 10 Gy {gamma}-rays or a fractionated dose of 40 Gy {gamma}-rays, total) or sham-irradiation and were maintained for 4, 8, and 24 h following irradiation. mRNA expression levels of MMPs and TIMPs in the brain were analyzed by real-time reverse transcriptase-polymerase chain reaction (PCR). The functional activity of MMPs was measured by in situ zymography, and degradation of ECM was visualized by collagen type IV immunofluorescent staining. Results: A significant increase in mRNA expression levels of MMP-2, MMP-9, and TIMP-1 was observed in irradiated brains compared to that in sham-irradiated controls. In situ zymography revealed a strong gelatinolytic activity in the brain 24 h postirradiation, and the enhanced gelatinolytic activity mediated by irradiation was significantly attenuated in the presence of anti-MMP-2 antibody. A significant reduction in collagen type IV immunoreactivity was also detected in the brain at 24 h after irradiation. In contrast, the levels of collagen type IV were not significantly changed at 4 and 8 h after irradiation compared with the sham-irradiated controls. Conclusions: The present study demonstrates for the first time that radiation induces an imbalance between MMP-2 and TIMP-2 levels and suggests that degradation of collagen type IV, a major ECM component of BBB basement membrane, may have a role in the pathogenesis of brain injury.

  19. Tensile strength of type IV dental stones dried in a microwave oven.

    Science.gov (United States)

    Hersek, Nur; Canay, Senay; Akça, Kivanç; Ciftçi, Yalçin

    2002-05-01

    It is known that drying dental stones in a microwave oven can save time, but the strength of the material may be affected by different drying methods. This study evaluated the diametral tensile strength (DTS) of 5 type IV gypsum products at different time intervals using microwave and air-drying methods. . A total of 300 cylinder specimens were prepared from 5 type IV dental stones (Moldano, Amberok, Herastone, Shera-Sockel, and Fujirock; n = 60 per stone) in accordance with the manufacturers' recommendations. Half of the specimens of each stone (n = 30) were dried in open air within a temperature range of 20 +/- 2 degrees C; the other half (n = 30) underwent initial setting in a silicone rubber mold in open air for 10 minutes and then were dried in a microwave oven for 10 minutes. Within these groups, 10 specimens were tested under diametral compression at each of the following time periods: 30, 60, and 120 minutes after drying. Three-way analysis of variance and Scheffe's post hoc test were performed for statistical comparisons at a significance level of Pmicrowave oven (mean 2.99 MPa) were significantly higher (Pmicrowave oven drying had a positive effect on the diametral tensile strength of 5 type IV dental stones.

  20. Alogliptin, a potent and selective dipeptidyl peptidase-IV inhibitor for the treatment of type 2 diabetes

    DEFF Research Database (Denmark)

    Deacon, Carolyn F

    2008-01-01

    Takeda San Diego Inc is developing alogliptin, a small-molecule, orally available dipeptidyl peptidase IV (DPP IV) inhibitor, for the potential treatment of type 2 diabetes. In January 2008, Takeda announced that an NDA for alogliptin had been submitted to the FDA.......Takeda San Diego Inc is developing alogliptin, a small-molecule, orally available dipeptidyl peptidase IV (DPP IV) inhibitor, for the potential treatment of type 2 diabetes. In January 2008, Takeda announced that an NDA for alogliptin had been submitted to the FDA....

  1. Confocal and conventional immunofluorescent and immunogold electron microscopic localization of collagen types III and IV in human placenta.

    Science.gov (United States)

    Nanaev, A K; Rukosuev, V S; Shirinsky, V P; Milovanov, A P; Domogatsky, S P; Duance, V C; Bradbury, F M; Yarrow, P; Gardiner, L; d'Lacey, C

    1991-01-01

    Confocal and conventional indirect immunofluorescence and immunogold electron microscopic methods were applied to examine the distribution of extracellular matrix constituents (collagens types III and IV) in the villi of immature and term human placentae. The immunofluorescence study revealed that collagen type III is more distinct in the villous stroma of term placenta as compared with that of the first trimester. Collagen type IV was detected mainly in endothelial and epithelial basement membranes and interestingly also to a certain extent in the stroma. Results obtained using immunoelectron microscopy support the proposal that collagen types III and IV are characteristic of stromal and basement membranes, respectively. Stromal collagen type IV is apparently localized in association with the interstitial types of collagen (I and III), in the villous stroma of term placenta.

  2. 10 CFR Appendix IV to Part 960 - Types of Information for the Nomination of Sites as Suitable for Characterization

    Science.gov (United States)

    2010-01-01

    ... 10 Energy 4 2010-01-01 2010-01-01 false Types of Information for the Nomination of Sites as Suitable for Characterization IV Appendix IV to Part 960 Energy DEPARTMENT OF ENERGY GENERAL GUIDELINES FOR... hydraulic properties of aquifers, confining units, and aquitards. • Potential areas and modes of recharge...

  3. Immunofluorescent localization of collagen types I, III, IV, V, fibronectin, laminin, entactin, and heparan sulphate proteoglycan in human immature placenta.

    Science.gov (United States)

    Rukosuev, V S

    1992-03-15

    The distribution of eight components of the extracellular matrix in immature human placenta was studied by an indirect immunofluorescence method with monospecific antibodies. In the stroma of the term chorionic villi, collagen types I, III, IV, V, and fibronectin formed a mesh of fibers and conglomerates. Heparan sulphate proteoglycan formed multiple conglomerates, whereas laminin comprised small, scanty, discrete granules. Collagen type IV, laminin, entactin, and heparan sulphate proteoglycan were confined to the basement membrane of the trophoblast. Sometimes, only collagen type IV was identified in fetal vascular basement membrane.

  4. The gonococcal genetic island and type IV secretion in the pathogenic Neisseria

    Directory of Open Access Journals (Sweden)

    Meghan E Ramsey

    2011-04-01

    Full Text Available Eighty percent of Neisseria gonorrhoeae strains and some Neisseria meningitidis strains encode a 57 kb gonococcal genetic island (GGI. The GGI was horizontally acquired and is inserted in the chromosome at the replication terminus. The GGI is flanked by direct repeats, and site-specific recombination at these sites results in excision of the GGI and may be responsible for its original acquisition. Although the role of the GGI in N. meningitidis is unclear, the GGI in N. gonorrhoeae encodes a type IV secretion system (T4SS. Type IV secretion systems are versatile multi-protein complexes and include both conjugation systems as well as effector systems that translocate either proteins or DNA-protein complexes. In N. gonorrhoeae, the T4SS secretes single-stranded chromosomal DNA into the extracellular milieu in a contact-independent manner. Importantly, the DNA secreted through the T4SS is effective in natural transformation and therefore contributes to the spread of genetic information through Neisseria populations. Mutagenesis experiments have identified genes for DNA secretion including those encoding putative structural components of the apparatus, peptidoglycanases which may act in assembly, and relaxosome components for processing the DNA and delivering it to the apparatus. The T4SS may also play a role in infection by N. gonorrhoeae. During intracellular infection, N. gonorrhoeae requires the Ton complex for iron acquisition and survival. However, N. gonorrhoeae strains that do not express the Ton complex can survive intracellularly if they express structural components of the T4SS. These data provide evidence that the T4SS is expressed during intracellular infection and suggest that the T4SS may provide an advantage for intracellular survival. Here we review our current understanding of how the GGI and type IV secretion affect natural transformation and pathogenesis in N. gonorrhoeae and N. meningitidis.

  5. Aeromonas spp. possess at least two distinct type IV pilus families.

    Science.gov (United States)

    Barnett, T C; Kirov, S M; Strom, M S; Sanderson, K

    1997-10-01

    Type IV pili have been purified from strains of most of the Aeromonas species associated with gastroenteritis (A. veronii biovar sobria, A. hydrophila, A. trota and A. caviae). They appear to be a related family (molecular mass of pilin 19 to 23 kDa) with a tendency to bundle-formation. Hence, we have designated them 'bundle-forming pili' (Bfp). A type IV pilus biogenesis gene cluster (tapABCD) recently cloned from a strain of A. hydrophila, however, encoded a 17 kDa pilin which differed significantly in its N-terminal amino acid sequence from the Bfp pilins. This paper describes the cloning of part (tapA and approximately 20% of tapB) of a homologous pilin gene cluster from a Bfp-positive strain of A. veronii biovar sobria, and presents evidence that the entire pilin gene cluster (tapABCD) is present in this strain. The predicted N-terminal amino acid sequence of the pilin encoded by the A. veronii biovar sobria tapA differed markedly from the corresponding sequence of its Bfp pilin, and those of the Bfp purified from other Aeromonas strains and species. Probing with tapA and tapD genes showed that these Bfp-positive Aeromonas strains also possessed the Tap gene cluster. TapA proteins of A. veronii biovar sobria and A. hydrophila shared 53% identity and 63% homology. We conclude that Aeromonas species are potentially able to express at least two distinct families of type IV pili (Bfp and Tap). Copyright 1997 Academic Press Limited.

  6. Pathological personality traits can capture DSM-IV personality disorder types.

    Science.gov (United States)

    Miller, Joshua D; Few, Lauren R; Lynam, Donald R; MacKillop, James

    2015-01-01

    The Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition (DSM-5) includes an alternative diagnostic approach to the assessment of personality disorders (PDs) in Section III with the aim of stimulating further research. Diagnosis of a PD using this approach is predicated on the presence of personality impairment and pathological personality traits. The types of traits present (e.g., callousness vs. emotional lability) are used to derive DSM-IV PD scores. Concerns have been raised, however, that such a trait-based approach will yield PD constructs that differ substantially from those generated using the approaches articulated in previous iterations of the DSM. We empirically examined this issue in a sample of 109 adults who were currently receiving mental health treatment. More specifically, we examined the correlations between interview-based PD scores derived from DSM-IV to DSM-5 PD trait counts, and tested them in relation to the 30 specific facets of the five-factor model, as well as internalizing and externalizing symptoms. Overall, the DSM-IV PD scores and DSM-5 PD trait counts correlated significantly with one another (Mr = .63), demonstrated similar patterns of interrelations among the PDs, and manifested highly similar patterns of correlations with general personality traits and symptoms of psychopathology. These results indicate that the DSM-5 PD trait counts specified in the alternative DSM-5 PD diagnostic approach capture the same constructs as those measured using the more traditional DSM-IV diagnostic system. (PsycINFO Database Record (c) 2015 APA, all rights reserved).

  7. Two types of lanthanide selenidostannates(IV) first prepared under the same solvothermal conditions.

    Science.gov (United States)

    Zhou, Jian; Xiao, Hong; Xiao, Hong-Ping; Yang, Tao; Zou, Hua-Hong; Liu, Xing; Zhao, Rong-Qing; Tang, Qiuling

    2015-01-21

    Two types of lanthanide selenidostannates(iv) [Ln2(tepa)2(μ-OH)2Sn2Se6] {Ln = Y(), Pr (), Dy (), Er (), Tm (); tepa = tetraethylenepentamine} and [Ln2(tepa)2(μ2-OH)2Cl2]2[Sn4Se10]·4H2O {Ln = Y (), Dy (), Er (), Tm ()} have been synthesized under identical solvothermal conditions and characterized structurally. Type I (, , , and ) displays 1-D neutral chains [Ln2(tepa)2(μ-OH)2Sn2Se6]n, while type II (, , and ) contains discrete adamantane-like [Sn4Se10](4-) ions with binuclear lanthanide complex [Ln2(tepa)2(μ-OH)2Cl2](2+) ions as counterions. Although the solvothermal synthetic methods could result in the formation of various transition-metal chalcogenidometalates, such identical experimental conditions usually result in the only stable phases of lanthanide chalcogenidometalates. Hence, two different lanthanide selenidostannates(iv), obtained under same solvothermal conditions and starting materials, have been first observed in this work. The optical properties of all the compounds have been investigated by UV-vis spectra.

  8. [Vascular and/or cardiac manifestations of type IV Ehlers-Danlos syndrome. 9 cases].

    Science.gov (United States)

    de Wazières, B; Coppere, B; Durieu, I; Fest, T; Ninet, J; Levrat, R; Vuitton, D A; Dupond, J L

    1995-10-14

    Type IV Ehlers-Danlos syndrome, a rare disease caused by abnormal synthesis of type III collagen, often leads to vascular fragility. We report 9 cases (6 men and 3 women, mean age 35 years). For 7 of the patients, the inaugural signs were arterial complications including haemoperitoneum in 2 patients with multiple aneurysmal dystrophy of the abdominal arteries, one case of ruptured subclavian artery, two dissections of the renal artery, one case with rupture of a cerebral aneurysm, one rupture of the mesenteric artery and a haematoma after arterial puncture. Other vascular manifestations were acrosyndrome (n = 4), varicose veins (n = 3), and prolapsus of the mitral valve (n = 2). In addition, 8 of the 9 patients presented extravascular signs. There was a history of familial disease in 5 cases. Pregnancy was completed to term in three patients: a cesarean section was required in one case and intra-uterine growth retardation was seen in 2. Morbidity was important with hemiparesia, blindness and paraparesis sequellae. One patient died from haemorrhage. This series of patients with type IV Ehlers-Danlos syndrome illustrates the severity of this disease whose prevalence is often underestimated. The disease is transmitted by autosomal dominant inheritance, underscoring the importance of familial testing for early diagnosis. Clinicians should be aware of the vascular manifestations and avoid invasive punctures or operations except in exceptional indications.

  9. Structural characterization of outer membrane components of the type IV pili system in pathogenic Neisseria.

    Directory of Open Access Journals (Sweden)

    Samta Jain

    Full Text Available Structures of the type IV pili secretin complexes from Neisseria gonorrhoeae and Neisseria meningitidis, embedded in outer membranes were investigated by transmission electron microscopy. Single particle averaging revealed additional domains not observed previously. Secretin complexes of N. gonorrhoeae showed a double ring structure with a 14-15-fold symmetry in the central ring, and a 14-fold symmetry of the peripheral ring with 7 spikes protruding. In secretin complexes of N. meningitidis, the spikes were absent and the peripheral ring was partly or completely lacking. When present, it had a 19-fold symmetry. The structures of the complexes in several pil mutants were determined. Structures obtained from the pilC1/C2 adhesin and the pilW minor pilin deletion strains were similar to wild-type, whereas deletion of the homologue of N. meningitidis PilW resulted in the absence of secretin structures. Remarkably, the pilE pilin subunit and pilP lipoprotein deletion mutants showed a change in the symmetry of the peripheral ring from 14 to 19 and loss of spikes. The pilF ATPase mutant also lost the spikes, but maintained 14-fold symmetry. These results show that secretin complexes contain previously unidentified large and flexible extra domains with a probable role in stabilization or assembly of type IV pili.

  10. A widespread bacteriophage abortive infection system functions through a Type IV toxin–antitoxin mechanism

    OpenAIRE

    Dy, Ron L.; Przybilski, Rita; Semeijn, Koen; Salmond, George P. C.; Fineran, Peter C.

    2014-01-01

    Bacterial abortive infection (Abi) systems are ‘altruistic’ cell death systems that are activated by phage infection and limit viral replication, thereby providing protection to the bacterial population. Here, we have used a novel approach of screening Abi systems as a tool to identify and characterize toxin–antitoxin (TA)-acting Abi systems. We show that AbiE systems are encoded by bicistronic operons and function via a non-interacting (Type IV) bacteriostatic TA mechanism. The abiE operon w...

  11. Systemic Correction of Murine Glycogen Storage Disease Type IV by an AAV-Mediated Gene Therapy.

    Science.gov (United States)

    Yi, Haiqing; Zhang, Quan; Brooks, Elizabeth D; Yang, Chunyu; Thurberg, Beth L; Kishnani, Priya S; Sun, Baodong

    2017-03-01

    Deficiency of glycogen branching enzyme (GBE) causes glycogen storage disease type IV (GSD IV), which is characterized by the accumulation of a less branched, poorly soluble form of glycogen called polyglucosan (PG) in multiple tissues. This study evaluates the efficacy of gene therapy with an adeno-associated viral (AAV) vector in a mouse model of adult form of GSD IV (Gbe1ys/ys). An AAV serotype 9 (AAV9) vector containing a human GBE expression cassette (AAV-GBE) was intravenously injected into 14-day-old Gbe1ys/ys mice at a dose of 5 × 1011 vector genomes per mouse. Mice were euthanized at 3 and 9 months of age. In the AAV-treated mice at 3 months of age, GBE enzyme activity was highly elevated in heart, which is consistent with the high copy number of the viral vector genome detected. GBE activity also increased significantly in skeletal muscles and the brain, but not in the liver. The glycogen content was reduced to wild-type levels in muscles and significantly reduced in the liver and brain. At 9 months of age, though GBE activity was only significantly elevated in the heart, glycogen levels were significantly reduced in the liver, brain, and skeletal muscles of the AAV-treated mice. In addition, the AAV treatment resulted in an overall decrease in plasma activities of alanine transaminase, aspartate transaminase, and creatine kinase, and a significant increase in fasting plasma glucose concentration at 9 months of age. This suggests an alleviation of damage and improvement of function in the liver and muscles by the AAV treatment. This study demonstrated a long-term benefit of a systemic injection of an AAV-GBE vector in Gbe1ys/ys mice.

  12. Internal carotid artery dissection in a patient with Ehlers-Danlos syndrome type IV: diagnosis and management

    Directory of Open Access Journals (Sweden)

    Michel Nasser

    2013-06-01

    Full Text Available Ehlers-Danlos syndrome (EDS type IV, also known as vascular EDS, is an inherited connective tissue disorder with an estimated prevalence of 1/100,000 to 1/250,000. In EDS type IV, vascular complications may affect all anatomical areas, with a preference for large- and medium-sized arteries. Dissections of the vertebral and carotid arteries in their extra- and intra-cranial segments are typical. The authors report the case of a patient with EDS type IV for whom the diagnosis was established based on clinical signs and who developed internal carotid artery dissection at the age of 44 years. In the absence of a specific treatment for EDS type IV, medical interventions should focus on symptomatic relief, prophylactic measures, and genetic counseling. Invasive imaging techniques are contraindicated, and a conservative approach to vascular complications is usually recommended.

  13. Structural features of Cas2 from Thermococcus onnurineus in CRISPR-cas system type IV.

    Science.gov (United States)

    Jung, Tae-Yang; Park, Kwang-Hyun; An, Yan; Schulga, Alexy; Deyev, Sergey; Jung, Jong-Hyun; Woo, Eui-Jeon

    2016-10-01

    CRISPR-Cas is RNA-based prokaryotic immune systems that defend against exogenous genetic elements such as plasmids and viruses. Cas1 and Cas2 are highly conserved components that play an essential part in the adaptation stage of all CRISPR-Cas systems. Characterization of CRISPR-Cas genes in Thermococcus onnurineus reveals the association of the Cas2 gene with the putative type IV system that lacks Cas1 or its homologous genes. Here, we present a crystal structure of T. onnurineus Cas2 (Ton_Cas2) that exhibits a deep and wide cleft at an interface lined with positive residues (Arg16, Lys18, Lys19, Arg22, and Arg23). The obvious DNA recognizing loops in Cas2 from E. coli (Eco_Cas2) are absent in Ton_Cas2 and have significantly different shapes and electrostatic potential distributions around the putative nucleotide binding region. Furthermore, Ton_Cas2 lacks the hairpin motif at the C-terminus that is responsible for Cas1 binding in Eco_Cas2. These structural features could be a unique signature and indicate an altered functional mechanism in the adaptation stage of Cas2 in type IV CRISPR-Cas systems. © 2016 The Protein Society.

  14. Biomechanical study of modular hemipelvic endoprosthesis for Type I-IV defect of pelvic tumor.

    Science.gov (United States)

    Dong, Yang; Hu, Hai; Zhang, Chang-Qing

    2014-08-01

    The modular hemipelvic prosthesis has been used in patient of Type I-IV pelvic tumor with good outcomes, but how to keep the stability between the prosthesis and the residual sacrum is a problem. An additional screw-rod system seems to solve it, but its biomechanical characters are still not well understood, which need experimental evaluation. Six pelvic specimens were prepared in three conditions (normal intact pelvis, "normal"; the pelvis of left Type I-IV defect and implanted with prosthesis without/with additional screw-rod system, "rod-" and "rod+"). Compressing biomechanical experiments (50-500N) were performed in these three conditions, respectively. The loadings during the experiments are in accordance with the linear elastic control mode. Under the increasing loading, the implanted pelvises displaced asymmetrically, unlike normal intact pelvis. The vertical displacement of "rod+" changed significantly, whereas "rod-" did not. For both implanted pelvis, right side displaced less than left side (P values <0.05). The implanted pelvis showed asymmetric displacement under loading, where healthy side displaced more. The implanted pelvis plus screw-rod system showed less displacement at implanted side but more at contralateral side in comparison with those without screw-rod system.

  15. Distinct activities of Bartonella henselae type IV secretion effector proteins modulate capillary-like sprout formation.

    Science.gov (United States)

    Scheidegger, F; Ellner, Y; Guye, P; Rhomberg, T A; Weber, H; Augustin, H G; Dehio, C

    2009-07-01

    The zoonotic pathogen Bartonella henselae (Bh) can lead to vasoproliferative tumour lesions in the skin and inner organs known as bacillary angiomatosis and bacillary peliosis. The knowledge on the molecular and cellular mechanisms involved in this pathogen-triggered angiogenic process is confined by the lack of a suitable animal model and a physiologically relevant cell culture model of angiogenesis. Here we employed a three-dimensional in vitro angiogenesis assay of collagen gel-embedded endothelial cell (EC) spheroids to study the angiogenic properties of Bh. Spheroids generated from Bh-infected ECs displayed a high capacity to form sprouts, which represent capillary-like projections into the collagen gel. The VirB/VirD4 type IV secretion system and a subset of its translocated Bartonella effector proteins (Beps) were found to profoundly modulate this Bh-induced sprouting activity. BepA, known to protect ECs from apoptosis, strongly promoted sprout formation. In contrast, BepG, triggering cytoskeletal rearrangements, potently inhibited sprouting. Hence, the here established in vitro model of Bartonella- induced angiogenesis revealed distinct and opposing activities of type IV secretion system effector proteins, which together with a VirB/VirD4-independent effect may control the angiogenic activity of Bh during chronic infection of the vasculature.

  16. Open angle glaucoma in a case of Type IV Ehler Danlos syndrome: A rarely reported association

    Directory of Open Access Journals (Sweden)

    Arijit Mitra

    2014-01-01

    Full Text Available A 26-year-old male presented to us with defective vision in the left eye. He had best corrected visual acuity (BCVA of hand movement (HM in right eye and 6/9 in left eye. He had ptosis with ectropion in both eyes and relative afferent pupillary defect (RAPD in right eye. Intraocular pressure (IOP was 46 and 44 mmHg in right and left eye, respectively. Fundus showed glaucomatous optic atrophy (GOA in right eye and cup disc ratio (CDR of 0.75 with bipolar rim thinning in left eye. Systemic examination showed hyperextensible skin and joints, acrogeria, hypodontia, high arched palate, and varicose veins. He gave history of easy bruising and tendency to fall and history of intestinal rupture 5 years ago for which he had undergone surgery. He was diagnosed as a case of Type IV Ehler-Danlos syndrome (EDS with open angle glaucoma. He underwent trabeculectomy in both eyes. This is a rare case that shows glaucoma in a patient of EDS Type IV. Very few such cases have been reported in literature.

  17. Molecular recognition determinants for type IV secretion of diverse families of conjugative relaxases.

    Science.gov (United States)

    Lang, Silvia; Gruber, Karl; Mihajlovic, Sanja; Arnold, Roland; Gruber, Christian J; Steinlechner, Sonja; Jehl, Marc-Andre; Rattei, Thomas; Fröhlich, Kai-Uwe; Zechner, Ellen L

    2010-12-01

    In preparation for transfer conjugative type IV secretion systems (T4SS) produce a nucleoprotein adduct containing a relaxase enzyme covalently linked to the 5' end of single-stranded plasmid DNA. The bound relaxase is expected to present features necessary for selective recognition by the type IV coupling protein (T4CP), which controls substrate entry to the envelope spanning secretion machinery. We prove that the IncF plasmid R1 relaxase TraI is translocated to the recipient cells. Using a Cre recombinase assay (CRAfT) we mapped two internally positioned translocation signals (TS) on F-like TraI proteins that independently mediate efficient recognition and secretion. Tertiary structure predictions for the TS matched best helicase RecD2 from Deinococcus radiodurans. The TS is widely conserved in MOB(F) and MOB(Q) families of relaxases. Structure/function relationships within the TS were identified by mutation. A key residue in specific recognition by T4CP TraD was revealed by a fidelity switch phenotype for an F to plasmid R1 exchange L626H mutation. Finally, we show that physical linkage of the relaxase catalytic domain to a TraI TS is necessary for efficient conjugative transfer. © 2010 Blackwell Publishing Ltd.

  18. Earthworm symbiont Verminephrobacter eiseniae mediates natural transformation within the host egg capsules using type IV pili

    Directory of Open Access Journals (Sweden)

    SEANA Kelyn DAVIDSON

    2014-10-01

    Full Text Available The dense microbial communities commonly associated with plants and animals should offer many opportunities for horizontal gene transfer (HGT through described mechanisms of DNA exchange including natural transformation. However, studies of the significance of natural transformation have focused primarily on pathogens. The study presented here demonstrates highly efficient DNA exchange by natural transformation in a common symbiont of earthworms. The obligate bacterial symbiont Verminephrobacter eiseniae is a member of a microbial consortium of the earthworm Eisenia fetida that is transmitted into the egg capsules to colonize the embryonic worms. In the study presented here, by testing for transformants under different conditions in culture, we demonstrate that V. eiseniae can incorporate free DNA from the environment, that competency is regulated by environmental factors, and that it is sequence specific. Mutations in the type IV pili of V. eiseniae resulted in loss of DNA uptake, implicating the type IV pilus (TFP apparatus in DNA uptake. Furthermore, injection of DNA carrying antibiotic-resistance genes into egg capsules resulted in transformants within the capsule, demonstrating the relevance of DNA uptake within the earthworm system. The ability to take up species-specific DNA from the environment may explain the maintenance of the relatively large, intact genome of this long-associated obligate symbiont, and provides a mechanism for acquisition of foreign genes within the earthworm system.

  19. Systematic screens for proteins that interact with the mucolipidosis type IV protein TRPML1.

    Directory of Open Access Journals (Sweden)

    Ellen Spooner

    Full Text Available Mucolipidosis type IV is a lysosomal storage disorder resulting from mutations in the MCOLN1 gene, which encodes the endosomal/lysosomal Transient Receptor Potential channel protein mucolipin-1/TRPML1. Cells isolated from Mucolipidosis type IV patients and grown in vitro and in in vivo models of this disease both show several lysosome-associated defects. However, it is still unclear how TRPML1 regulates the transport steps implicated by these defects. Identifying proteins that associate with TRPML1 will facilitate the elucidation of its cellular and biochemical functions. We report here two saturation screens for proteins that interact with TRPML1: one that is based on immunoprecipitation/mass spectrometry and the other using a genetic yeast two-hybrid approach. From these screens, we identified largely non-overlapping proteins, which represent potential TRPML1-interactors., Using additional interaction assays on some of the potential interactors from each screen, we validated some proteins as candidate TRPML1 interactors In addition, our analysis indicates that each of the two screens not only identified some false-positive interactors, as expected from any screen, but also failed to uncover potential TRPML1 interactors. Future studies on the true interactors, first identified in these screens, will help elucidate the structure and function of protein complexes containing TRPML1.

  20. Methods to Improve Osseointegration of Dental Implants in Low Quality (Type-IV Bone: An Overview

    Directory of Open Access Journals (Sweden)

    Hamdan S. Alghamdi

    2018-01-01

    Full Text Available Nowadays, dental implants have become more common treatment for replacing missing teeth and aim to improve chewing efficiency, physical health, and esthetics. The favorable clinical performance of dental implants has been attributed to their firm osseointegration, as introduced by Brånemark in 1965. Although the survival rate of dental implants over a 10-year observation has been reported to be higher than 90% in totally edentulous jaws, the clinical outcome of implant treatment is challenged in compromised (bone conditions, as are frequently present in elderly people. The biomechanical characteristics of bone in aged patients do not offer proper stability to implants, being similar to type-IV bone (Lekholm & Zarb classification, in which a decreased clinical fixation of implants has been clearly demonstrated. However, the search for improved osseointegration has continued forward for the new evolution of modern dental implants. This represents a continuum of developments spanning more than 20 years of research on implant related-factors including surgical techniques, implant design, and surface properties. The methods to enhance osseointegration of dental implants in low quality (type-IV bone are described in a general manner in this review.

  1. Methods to Improve Osseointegration of Dental Implants in Low Quality (Type-IV) Bone: An Overview.

    Science.gov (United States)

    Alghamdi, Hamdan S

    2018-01-13

    Nowadays, dental implants have become more common treatment for replacing missing teeth and aim to improve chewing efficiency, physical health, and esthetics. The favorable clinical performance of dental implants has been attributed to their firm osseointegration, as introduced by Brånemark in 1965. Although the survival rate of dental implants over a 10-year observation has been reported to be higher than 90% in totally edentulous jaws, the clinical outcome of implant treatment is challenged in compromised (bone) conditions, as are frequently present in elderly people. The biomechanical characteristics of bone in aged patients do not offer proper stability to implants, being similar to type-IV bone (Lekholm & Zarb classification), in which a decreased clinical fixation of implants has been clearly demonstrated. However, the search for improved osseointegration has continued forward for the new evolution of modern dental implants. This represents a continuum of developments spanning more than 20 years of research on implant related-factors including surgical techniques, implant design, and surface properties. The methods to enhance osseointegration of dental implants in low quality (type-IV) bone are described in a general manner in this review.

  2. Identification of a Novel Conjugative Plasmid in Mycobacteria That Requires Both Type IV and Type VII Secretion

    KAUST Repository

    Ummels, R.

    2014-09-23

    Conjugative plasmids have been identified in a wide variety of different bacteria, ranging from proteobacteria to firmicutes, and conjugation is one of the most efficient routes for horizontal gene transfer. The most widespread mechanism of plasmid conjugation relies on different variants of the type IV secretion pathway. Here, we describe the identification of a novel type of conjugative plasmid that seems to be unique for mycobacteria. Interestingly, while this plasmid is efficiently exchanged between different species of slow-growing mycobacteria, including Mycobacterium tuberculosis, it could not be transferred to any of the fast-growing mycobacteria tested. Genetic analysis of the conjugative plasmid showed the presence of a locus containing homologues of three type IV secretion system components and a relaxase. In addition, a new type VII secretion locus was present. Using transposon insertion mutagenesis, we show that in fact both these secretion systems are essential for conjugation, indicating that this plasmid represents a new class of conjugative plasmids requiring two secretion machineries. This plasmid could form a useful new tool to exchange or introduce DNA in slow-growing mycobacteria. IMPORTANCE: Conjugative plasmids play an important role in horizontal gene transfer between different bacteria and, as such, in their adaptation and evolution. This effect is most obvious in the spread of antibiotic resistance genes. Thus far, conjugation of natural plasmids has been described only rarely for mycobacterial species. In fact, it is generally accepted that M. tuberculosis does not show any recent sign of horizontal gene transfer. In this study, we describe the identification of a new widespread conjugative plasmid that can also be efficiently transferred to M. tuberculosis. This plasmid therefore poses both a threat and an opportunity. The threat is that, through the acquisition of antibiotic resistance markers, this plasmid could start a rapid spread of

  3. Attenuation of the Type IV Pilus Retraction Motor Influences Neisseria gonorrhoeae Social and Infection Behavior.

    Science.gov (United States)

    Hockenberry, Alyson M; Hutchens, Danielle M; Agellon, Al; So, Magdalene

    2016-12-06

    Retraction of the type IV pilus (Tfp) mediates DNA uptake, motility, and social and infection behavior in a wide variety of prokaryotes. To date, investigations into Tfp retraction-dependent activities have used a mutant deleted of PilT, the ATPase motor protein that causes the pilus fiber to retract. ΔpilT cells are nontransformable, nonmotile, and cannot aggregate into microcolonies. We tested the hypothesis that these retraction-dependent activities are sensitive to the strength of PilT enzymatic activity by using the pathogen Neisseria gonorrhoeae as a model. We constructed an N. gonorrhoeae mutant with an amino acid substitution in the PilT Walker B box (a substitution of cysteine for leucine at position 201, encoded by pilTL201C). Purified PilTL201C forms a native hexamer, but mutant hexamers hydrolyze ATP at half the maximal rate. N. gonorrhoeae pilTL201C cells produce Tfp fibers, crawl at the same speed as the wild-type (wt) parent, and are equally transformable. However, the social behavior of pilTL201C cells is intermediate between the behaviors of wt and ΔpilT cells. The infection behavior of pilTL201C is also defective, due to its failure to activate the epidermal growth factor receptor (EGFR)-heparin-binding EGF-like growth factor (HB-EGF) pathway. Our study indicates that pilus retraction, per se, is not sufficient for N. gonorrhoeae microcolony formation or infectivity; rather, these activities are sensitive to the strength of PilT enzymatic activity. We discuss the implications of these findings for Neisseria pathogenesis in the context of mechanobiology. Type IV pili are fibers expressed on the surface of many bacteria. Neisseria gonorrhoeae cells crawl, take up DNA, and communicate with each other and with human cells by retracting these fibers. Here, we show that an N. gonorrhoeae mutant expressing an enzymatically weakened type IV pilus retraction motor still crawls and takes up DNA normally. However, mutant cells exhibit abnormal social

  4. Systematic analysis and prediction of type IV secreted effector proteins by machine learning approaches.

    Science.gov (United States)

    Wang, Jiawei; Yang, Bingjiao; An, Yi; Marquez-Lago, Tatiana; Leier, André; Wilksch, Jonathan; Hong, Qingyang; Zhang, Yang; Hayashida, Morihiro; Akutsu, Tatsuya; Webb, Geoffrey I; Strugnell, Richard A; Song, Jiangning; Lithgow, Trevor

    2017-11-27

    In the course of infecting their hosts, pathogenic bacteria secrete numerous effectors, namely, bacterial proteins that pervert host cell biology. Many Gram-negative bacteria, including context-dependent human pathogens, use a type IV secretion system (T4SS) to translocate effectors directly into the cytosol of host cells. Various type IV secreted effectors (T4SEs) have been experimentally validated to play crucial roles in virulence by manipulating host cell gene expression and other processes. Consequently, the identification of novel effector proteins is an important step in increasing our understanding of host-pathogen interactions and bacterial pathogenesis. Here, we train and compare six machine learning models, namely, Naïve Bayes (NB), K-nearest neighbor (KNN), logistic regression (LR), random forest (RF), support vector machines (SVMs) and multilayer perceptron (MLP), for the identification of T4SEs using 10 types of selected features and 5-fold cross-validation. Our study shows that: (1) including different but complementary features generally enhance the predictive performance of T4SEs; (2) ensemble models, obtained by integrating individual single-feature models, exhibit a significantly improved predictive performance and (3) the 'majority voting strategy' led to a more stable and accurate classification performance when applied to predicting an ensemble learning model with distinct single features. We further developed a new method to effectively predict T4SEs, Bastion4 (Bacterial secretion effector predictor for T4SS), and we show our ensemble classifier clearly outperforms two recent prediction tools. In summary, we developed a state-of-the-art T4SE predictor by conducting a comprehensive performance evaluation of different machine learning algorithms along with a detailed analysis of single- and multi-feature selections. © The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  5. A widespread bacteriophage abortive infection system functions through a Type IV toxin-antitoxin mechanism.

    Science.gov (United States)

    Dy, Ron L; Przybilski, Rita; Semeijn, Koen; Salmond, George P C; Fineran, Peter C

    2014-04-01

    Bacterial abortive infection (Abi) systems are 'altruistic' cell death systems that are activated by phage infection and limit viral replication, thereby providing protection to the bacterial population. Here, we have used a novel approach of screening Abi systems as a tool to identify and characterize toxin-antitoxin (TA)-acting Abi systems. We show that AbiE systems are encoded by bicistronic operons and function via a non-interacting (Type IV) bacteriostatic TA mechanism. The abiE operon was negatively autoregulated by the antitoxin, AbiEi, a member of a widespread family of putative transcriptional regulators. AbiEi has an N-terminal winged-helix-turn-helix domain that is required for repression of abiE transcription, and an uncharacterized bi-functional C-terminal domain, which is necessary for transcriptional repression and sufficient for toxin neutralization. The cognate toxin, AbiEii, is a predicted nucleotidyltransferase (NTase) and member of the DNA polymerase β family. AbiEii specifically bound GTP, and mutations in conserved NTase motifs (I-III) and a newly identified motif (IV), abolished GTP binding and subsequent toxicity. The AbiE systems can provide phage resistance and enable stabilization of mobile genetic elements, such as plasmids. Our study reveals molecular insights into the regulation and function of the widespread bi-functional AbiE Abi-TA systems and the biochemical properties of both toxin and antitoxin proteins.

  6. A widespread bacteriophage abortive infection system functions through a Type IV toxin–antitoxin mechanism

    Science.gov (United States)

    Dy, Ron L.; Przybilski, Rita; Semeijn, Koen; Salmond, George P.C.; Fineran, Peter C.

    2014-01-01

    Bacterial abortive infection (Abi) systems are ‘altruistic’ cell death systems that are activated by phage infection and limit viral replication, thereby providing protection to the bacterial population. Here, we have used a novel approach of screening Abi systems as a tool to identify and characterize toxin–antitoxin (TA)-acting Abi systems. We show that AbiE systems are encoded by bicistronic operons and function via a non-interacting (Type IV) bacteriostatic TA mechanism. The abiE operon was negatively autoregulated by the antitoxin, AbiEi, a member of a widespread family of putative transcriptional regulators. AbiEi has an N-terminal winged-helix-turn-helix domain that is required for repression of abiE transcription, and an uncharacterized bi-functional C-terminal domain, which is necessary for transcriptional repression and sufficient for toxin neutralization. The cognate toxin, AbiEii, is a predicted nucleotidyltransferase (NTase) and member of the DNA polymerase β family. AbiEii specifically bound GTP, and mutations in conserved NTase motifs (I-III) and a newly identified motif (IV), abolished GTP binding and subsequent toxicity. The AbiE systems can provide phage resistance and enable stabilization of mobile genetic elements, such as plasmids. Our study reveals molecular insights into the regulation and function of the widespread bi-functional AbiE Abi-TA systems and the biochemical properties of both toxin and antitoxin proteins. PMID:24465005

  7. Participation of collagen types I, III, IV, V, and fibronectin in the formation of villi fibrosis in human term placenta.

    Science.gov (United States)

    Rukosuev, V S; Nanaev, A K; Milovanov, A P

    1990-01-01

    The indirect immunofluorescence method was used to study the human term placenta in pathological pregnancy for the distribution of collagen types I, III, IV, V, and fibronectin in fibrosis stromatis villi. All collagen types and fibronectin were shown to participate in fibrosis villorum formation. Fibronectin was also detected in the fibrinoid that surrounded villi at stroma. The presence of free cytotrophoblast cells in the fibrinoid was accompanied by a noticeable increase in fibronectin fluorescence. A significant amount of collagen types IV and V and a less amount of collagen types I and III were identified.

  8. Longevity of dental implants in type IV bone: a systematic review.

    Science.gov (United States)

    Goiato, M C; dos Santos, D M; Santiago, J F; Moreno, A; Pellizzer, E P

    2014-09-01

    Bone quality and quantity are important factors with regard to the survival rate of dental implants. The aim of this study was to conduct a systematic review of dental implants inserted in low-density bone and to determine the survival rate of dental implants with surface treatments over time. A systematic review of the literature was undertaken by two independent individuals; the Medline/PubMed database was searched for the period July 1975 to March 2013. Relevant reports on bone quality and osseointegration of dental implants were selected. The search retrieved 1018 references, and after inclusion and exclusion criteria were applied, 19 studies were selected for review. A total of 3937 patients, who had received a total of 12,465 dental implants, were analyzed. The survival rates of dental implants according to the bone density were: type I, 97.6%; type II, 96.2%; type III, 96.5%; and type IV, 88.8%. The survival rate of treated surface implants inserted in low-density bone was higher (97.1%) than that of machined surface implants (91.6%). Surface-treated dental implants inserted in low-density bone have a high survival rate and may be indicated for oral rehabilitation. However, more randomized studies are required to better evaluate this issue. Copyright © 2014 International Association of Oral and Maxillofacial Surgeons. Published by Elsevier Ltd. All rights reserved.

  9. A type IV pilus mediates DNA binding during natural transformation in Streptococcus pneumoniae.

    Directory of Open Access Journals (Sweden)

    Raphaël Laurenceau

    Full Text Available Natural genetic transformation is widely distributed in bacteria and generally occurs during a genetically programmed differentiated state called competence. This process promotes genome plasticity and adaptability in Gram-negative and Gram-positive bacteria. Transformation requires the binding and internalization of exogenous DNA, the mechanisms of which are unclear. Here, we report the discovery of a transformation pilus at the surface of competent Streptococcus pneumoniae cells. This Type IV-like pilus, which is primarily composed of the ComGC pilin, is required for transformation. We provide evidence that it directly binds DNA and propose that the transformation pilus is the primary DNA receptor on the bacterial cell during transformation in S. pneumoniae. Being a central component of the transformation apparatus, the transformation pilus enables S. pneumoniae, a major Gram-positive human pathogen, to acquire resistance to antibiotics and to escape vaccines through the binding and incorporation of new genetic material.

  10. Reconstitution of a minimal machinery capable of assembling periplasmic type IV pili.

    Science.gov (United States)

    Goosens, Vivianne J; Busch, Andreas; Georgiadou, Michaella; Castagnini, Marta; Forest, Katrina T; Waksman, Gabriel; Pelicic, Vladimir

    2017-06-20

    Type IV pili (Tfp), which are key virulence factors in many bacterial pathogens, define a large group of multipurpose filamentous nanomachines widespread in Bacteria and Archaea. Tfp biogenesis is a complex multistep process, which relies on macromolecular assemblies composed of 15 conserved proteins in model gram-negative species. To improve our limited understanding of the molecular mechanisms of filament assembly, we have used a synthetic biology approach to reconstitute, in a nonnative heterologous host, a minimal machinery capable of building Tfp. Here we show that eight synthetic genes are sufficient to promote filament assembly and that the corresponding proteins form a macromolecular complex at the cytoplasmic membrane, which we have purified and characterized biochemically. Our results contribute to a better mechanistic understanding of the assembly of remarkable dynamic filaments nearly ubiquitous in prokaryotes.

  11. Type IV hiatal hernia post laparoscopic Nissen fundoplication: report of a case.

    LENUS (Irish Health Repository)

    Awad, Z T

    2001-01-01

    A postoperative hiatal hernia is a rare but serious complication of fundoplication. We report herein a 62-year-old female who presented with abdominal pain and vomiting 2 years following laparoscopic Nissen fundoplication. At laparotomy, the stomach and the transverse colon were intrathoracic (type IV hiatal hernia); the esophageal hiatus was markedly dilated with no evidence that they had been approximated. At 18 months follow-up, she is doing very well apart from occasional heartburn. A high index of suspicion is needed to diagnose postoperative hiatal hernias. A routine closure of the crura with nonabsorbable suture material and an avoidance of iatrogenic pneumothorax may help to reduce the occurrence of this problem.

  12. A model to predict the permeation of type IV hydrogen tanks

    Energy Technology Data Exchange (ETDEWEB)

    Bayle, Julien; Perreux, Dominique; Chapelle, David; Thiebaud, Frederic [MaHyTec, Dole (France); Nardin, Philippe [Franche Comte Univ. (France)

    2010-07-01

    In the frame of the certification process of the type IV hydrogen storage tanks MaHyTec aims to manufacture, this innovative SME is developing a numerical model dedicated to the study of permeation issues. Such an approach aims at avoiding complicated, time-consuming and expensive testing. Experimental results obtained under real conditions can moreover be significantly influenced by the scattering of material properties and liner dimensions. From simple testing on small-size flat membranes, the model allows to predict the gas diffusion flow through the whole structure by means of numerous parameters. On every step, theory can be compared with the results obtained from the samples. This document presents a brief review of the mathematical theory describing gas diffusion and the different aspects of the study for better understanding the proposed approach. (orig.)

  13. Type IV secretion systems and genomic islands-mediated horizontal gene transfer in Pseudomonas and Haemophilus.

    Science.gov (United States)

    Juhas, Mario

    2015-01-01

    Bacterial secretion systems, such as type IV secretion systems (T4SSs) are multi-subunit machines transferring macromolecules across membranes. Besides proteins, T4SSs also transfer nucleoprotein complexes, thus having a significant impact on the evolution of bacterial species. By T4SS-mediated horizontal gene transfer bacteria can acquire a broad spectrum of fitness genes allowing them to thrive in the wide variety of environments. Furthermore, acquisition of antibiotic-resistance and virulence genes can lead to the emergence of novel 'superbugs'. This review provides an update on the investigation of T4SSs. It highlights the role T4SSs play in the horizontal gene transfer, particularly in the evolution of catabolic pathways, antibiotic-resistance and virulence in Haemophilus and Pseudomonas. Copyright © 2014 Elsevier GmbH. All rights reserved.

  14. A type IV pilus mediates DNA binding during natural transformation in Streptococcus pneumoniae.

    Science.gov (United States)

    Laurenceau, Raphaël; Péhau-Arnaudet, Gérard; Baconnais, Sonia; Gault, Joseph; Malosse, Christian; Dujeancourt, Annick; Campo, Nathalie; Chamot-Rooke, Julia; Le Cam, Eric; Claverys, Jean-Pierre; Fronzes, Rémi

    2013-01-01

    Natural genetic transformation is widely distributed in bacteria and generally occurs during a genetically programmed differentiated state called competence. This process promotes genome plasticity and adaptability in Gram-negative and Gram-positive bacteria. Transformation requires the binding and internalization of exogenous DNA, the mechanisms of which are unclear. Here, we report the discovery of a transformation pilus at the surface of competent Streptococcus pneumoniae cells. This Type IV-like pilus, which is primarily composed of the ComGC pilin, is required for transformation. We provide evidence that it directly binds DNA and propose that the transformation pilus is the primary DNA receptor on the bacterial cell during transformation in S. pneumoniae. Being a central component of the transformation apparatus, the transformation pilus enables S. pneumoniae, a major Gram-positive human pathogen, to acquire resistance to antibiotics and to escape vaccines through the binding and incorporation of new genetic material.

  15. Evaluation of the Various Drying Methods on Surface Hardness of Type IV Dental Stone

    Science.gov (United States)

    Sudhakar, A; Srivatsa, G; Shetty, Rohit; Rajeswari, C L; Manvi, Supriya

    2015-01-01

    Background: Studies regarding the effect of various methods to increase the surface hardness of Type IV dental stone are not conclusive. Therefore, this study was carried out to evaluate the effect of air drying, micro oven drying and die hardener on surface hardness of Type IV dental stone. Materials and Methods: A standard metal die was fabricated; polyvinyl siloxane impression material was used to make the molds of metal die. A total of 120 specimens were obtained from two different die stones and were grouped as Group A (kalrock) and Group B (pearl stone), and were subjected to air drying for 24 h, micro oven drying and application of die hardener. These models were then subjected to surface hardness testing using the knoop hardness instrument. The obtained data were subjected to statistical analysis. Results: The hardness of Group A specimens was 64 ± 0.54 Knoop hardness number (KHN) after application of die hardener, 60.47 ± 0.41 KHN after 24 h air drying, 58.2 ± 0.88 after microwave oven drying and 24.6 ± 0.4 after 1 h air drying. The hardness of Group B specimens was 45.59 ± 0.63 KHN after application of die hardener, 40.2 ± 0.63 KHN after 24 h air drying, 38.28 ± 0.55 KHN after microwave oven drying and 19.91 ± 0.64 KHN after 1 h air drying. Conclusion: Group A showed better results than Group B at all times. Application of the die hardener showed highest hardness values followed in the order by 24 h air drying, microwave oven drying and 1 h air drying in both groups. The study showed that air drying the dies for 24 h followed by application of a single layer of the die hardener produced the best surface hardness and is recommended to be followed in practice. PMID:26124610

  16. Evaluation of the Various Drying Methods on Surface Hardness of Type IV Dental Stone.

    Science.gov (United States)

    Sudhakar, A; Srivatsa, G; Shetty, Rohit; Rajeswari, C L; Manvi, Supriya

    2015-06-01

    Studies regarding the effect of various methods to increase the surface hardness of Type IV dental stone are not conclusive. Therefore, this study was carried out to evaluate the effect of air drying, micro oven drying and die hardener on surface hardness of Type IV dental stone. A standard metal die was fabricated; polyvinyl siloxane impression material was used to make the molds of metal die. A total of 120 specimens were obtained from two different die stones and were grouped as Group A (kalrock) and Group B (pearl stone), and were subjected to air drying for 24 h, micro oven drying and application of die hardener. These models were then subjected to surface hardness testing using the knoop hardness instrument. The obtained data were subjected to statistical analysis. The hardness of Group A specimens was 64 ± 0.54 Knoop hardness number (KHN) after application of die hardener, 60.47 ± 0.41 KHN after 24 h air drying, 58.2 ± 0.88 after microwave oven drying and 24.6 ± 0.4 after 1 h air drying. The hardness of Group B specimens was 45.59 ± 0.63 KHN after application of die hardener, 40.2 ± 0.63 KHN after 24 h air drying, 38.28 ± 0.55 KHN after microwave oven drying and 19.91 ± 0.64 KHN after 1 h air drying. Group A showed better results than Group B at all times. Application of the die hardener showed highest hardness values followed in the order by 24 h air drying, microwave oven drying and 1 h air drying in both groups. The study showed that air drying the dies for 24 h followed by application of a single layer of the die hardener produced the best surface hardness and is recommended to be followed in practice.

  17. Reassessing the role of DotF in the Legionella pneumophila type IV secretion system.

    Directory of Open Access Journals (Sweden)

    Molly C Sutherland

    Full Text Available Legionella pneumophila, the causative agent of a severe pneumonia termed Legionnaires' Disease, survives and replicates within both protozoan hosts and human alveolar macrophages. Intracellular survival is dependent upon secretion of a plethora of protein effectors that function to form a replicative vacuole, evade the endocytic pathway and subvert host immune defenses. Export of these factors requires a type IV secretion system (T4SS called Dot/Icm that is composed of twenty-seven proteins. This report focuses on the DotF protein, which was previously postulated to have several different functions, one of which centered on binding Dot/Icm substrates. In this report, we examined if DotF functions as the T4SS inner membrane receptor for Dot/Icm substrates. Although we were able to recapitulate the previously published bacterial two-hybrid interaction between DotF and several substrates, the interaction was not dependent on the Dot/Icm substrates' signal sequences as predicted for a substrate:receptor interaction. In addition, binding did not require the cytoplasmic domain of DotF, which was anticipated to be involved in recognizing substrates in the cytoplasm. Finally, inactivation of dotF did not abolish intracellular growth of L. pneumophila or translocation of substrates, two phenotypes dependent on the T4SS receptor. These data strongly suggest that DotF does not act as the major receptor for Dot/Icm substrates and therefore likely performs an accessory function within the core-transmembrane subcomplex of the L. pneumophila Dot/Icm type IV secretion system.

  18. Methanosphaera stadtmanae induces a type IV hypersensitivity response in a mouse model of airway inflammation.

    Science.gov (United States)

    Bernatchez, Emilie; Gold, Matthew J; Langlois, Anick; Blais-Lecours, Pascale; Boucher, Magali; Duchaine, Caroline; Marsolais, David; McNagny, Kelly M; Blanchet, Marie-Renée

    2017-04-01

    Despite improved awareness of work-related diseases and preventive measures, many workers are still at high risk of developing occupational hypersensitivity airway diseases. This stems from a lack of knowledge of bioaerosol composition and their potential effects on human health. Recently, archaea species were identified in bioaerosols, raising the possibility that they play a major role in exposure-related pathology. Specifically, Methanosphaera stadtmanae (MSS) and Methanobrevibacter smithii (MBS) are found in high concentrations in agricultural environments and respiratory exposure to crude extract demonstrates immunomodulatory activity in mice. Nevertheless, our knowledge of the specific impact of methanogens exposure on airway immunity and their potential to induce airway hypersensitivity responses in workers remains scant. Analysis of the lung mucosal response to methanogen crude extracts in mice demonstrated that MSS and MBS predominantly induced TH17 airway inflammation, typical of a type IV hypersensitivity response. Furthermore, the response to MSS was associated with antigen-specific IgG1 and IgG2a production. However, despite the presence of eosinophils after MSS exposure, only a weak TH2 response and no airway hyperresponsiveness were observed. Finally, using eosinophil and mast cell-deficient mice, we confirmed that these cells are dispensable for the TH17 response to MSS, although eosinophils likely contribute to the exacerbation of inflammatory processes induced by MSS crude extract exposure. We conclude that, as MSS induces a clear type IV hypersensitivity lung response, it has the potential to be harmful to workers frequently exposed to this methanogen, and that preventive measures should be taken to avoid chronic hypersensitivity disease development in workers. © 2017 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of The Physiological Society and the American Physiological Society.

  19. Treatment of type 2 diabetes mellitus with agonists of the GLP-1 receptor or DPP-IV inhibitors

    DEFF Research Database (Denmark)

    Holst, Jens Juul

    2004-01-01

    in the treatment of Type 2 diabetes, causing marked improvements in glycaemic profile, insulin sensitivity and beta-cell performance, as well as weight reduction. The hormone is metabolised rapidly by the enzyme dipeptidyl peptidase IV (DPP-IV) and, therefore, cannot be easily used clinically. Instead, resistant...... with exendin have been carried out for > 6 months and have indicated efficacy in patients inadequately treated with oral antidiabetic agents. Orally active DPP-IV inhibitors, suitable for once-daily administration, have demonstrated similar efficacy. Diabetes therapy, based on GLP-1 receptor activation...

  20. AtlasT4SS: a curated database for type IV secretion systems.

    Science.gov (United States)

    Souza, Rangel C; del Rosario Quispe Saji, Guadalupe; Costa, Maiana O C; Netto, Diogo S; Lima, Nicholas C B; Klein, Cecília C; Vasconcelos, Ana Tereza R; Nicolás, Marisa F

    2012-08-09

    The type IV secretion system (T4SS) can be classified as a large family of macromolecule transporter systems, divided into three recognized sub-families, according to the well-known functions. The major sub-family is the conjugation system, which allows transfer of genetic material, such as a nucleoprotein, via cell contact among bacteria. Also, the conjugation system can transfer genetic material from bacteria to eukaryotic cells; such is the case with the T-DNA transfer of Agrobacterium tumefaciens to host plant cells. The system of effector protein transport constitutes the second sub-family, and the third one corresponds to the DNA uptake/release system. Genome analyses have revealed numerous T4SS in Bacteria and Archaea. The purpose of this work was to organize, classify, and integrate the T4SS data into a single database, called AtlasT4SS - the first public database devoted exclusively to this prokaryotic secretion system. The AtlasT4SS is a manual curated database that describes a large number of proteins related to the type IV secretion system reported so far in Gram-negative and Gram-positive bacteria, as well as in Archaea. The database was created using the RDBMS MySQL and the Catalyst Framework based in the Perl programming language and using the Model-View-Controller (MVC) design pattern for Web. The current version holds a comprehensive collection of 1,617 T4SS proteins from 58 Bacteria (49 Gram-negative and 9 Gram-Positive), one Archaea and 11 plasmids. By applying the bi-directional best hit (BBH) relationship in pairwise genome comparison, it was possible to obtain a core set of 134 clusters of orthologous genes encoding T4SS proteins. In our database we present one way of classifying orthologous groups of T4SSs in a hierarchical classification scheme with three levels. The first level comprises four classes that are based on the organization of genetic determinants, shared homologies, and evolutionary relationships: (i) F-T4SS, (ii) P-T4SS, (iii

  1. AtlasT4SS: A curated database for type IV secretion systems

    Directory of Open Access Journals (Sweden)

    Souza Rangel C

    2012-08-01

    Full Text Available Abstract Background The type IV secretion system (T4SS can be classified as a large family of macromolecule transporter systems, divided into three recognized sub-families, according to the well-known functions. The major sub-family is the conjugation system, which allows transfer of genetic material, such as a nucleoprotein, via cell contact among bacteria. Also, the conjugation system can transfer genetic material from bacteria to eukaryotic cells; such is the case with the T-DNA transfer of Agrobacterium tumefaciens to host plant cells. The system of effector protein transport constitutes the second sub-family, and the third one corresponds to the DNA uptake/release system. Genome analyses have revealed numerous T4SS in Bacteria and Archaea. The purpose of this work was to organize, classify, and integrate the T4SS data into a single database, called AtlasT4SS - the first public database devoted exclusively to this prokaryotic secretion system. Description The AtlasT4SS is a manual curated database that describes a large number of proteins related to the type IV secretion system reported so far in Gram-negative and Gram-positive bacteria, as well as in Archaea. The database was created using the RDBMS MySQL and the Catalyst Framework based in the Perl programming language and using the Model-View-Controller (MVC design pattern for Web. The current version holds a comprehensive collection of 1,617 T4SS proteins from 58 Bacteria (49 Gram-negative and 9 Gram-Positive, one Archaea and 11 plasmids. By applying the bi-directional best hit (BBH relationship in pairwise genome comparison, it was possible to obtain a core set of 134 clusters of orthologous genes encoding T4SS proteins. Conclusions In our database we present one way of classifying orthologous groups of T4SSs in a hierarchical classification scheme with three levels. The first level comprises four classes that are based on the organization of genetic determinants, shared homologies, and

  2. Abnormal deposition of laminin and type IV collagen at corneal epithelial basement membrane during wound healing in diabetic rats.

    Science.gov (United States)

    Sato, N; Nakamura, M; Chikama, T; Nishida, T

    1999-01-01

    To understand the pathophysiology of the corneal basement membrane in diabetes, we compared the localization of laminin and type IV collagen in the epithelial basement membrane during corneal epithelial wound healing in diabetic and nondiabetic rats. Streptozotocin was used to induce diabetes in half the rats. Two weeks later, the whole corneal epithelium was debrided. Diabetic and healthy rats (3-5 per group) were sacrificed before debridement and 1, 3, and 7 days and 1 month afterwards. The localization of laminin and type IV collagen was observed in cryosections by epifluorescence microscopy. In unwounded corneas of both diabetic and normal rats, laminin and type IV collagen were localized in the corneal epithelial basement. The intensity of fluorescence, however, was clearly stronger in the diabetic rats. In normal rats, wounding initially removed laminin and type IV collagen, but during healing these two proteins reappeared beneath the resurfacing corneal epithelium. Although similar results were observed in diabetic rats, the expression of laminin and type IV collagen was delayed, and their deposition was fragmented and irregular. These results suggest that delayed corneal epithelial wound healing in diabetes might involve delayed reappearance and abnormal reformation of epithelial basement membrane proteins.

  3. Plumbagin Alleviates Capillarization of Hepatic Sinusoids In Vitro by Downregulating ET-1, VEGF, LN, and Type IV Collagen

    Directory of Open Access Journals (Sweden)

    Guiyu Li

    2017-01-01

    Full Text Available Critical roles for liver sinusoidal endothelial cells (LSECs in liver fibrosis have been demonstrated, while little is known regarding the underlying molecular mechanisms of drugs delivered to the LSECs. Our previous study revealed that plumbagin plays an antifibrotic role in liver fibrosis. In this study, we investigated whether plumbagin alleviates capillarization of hepatic sinusoids by downregulating endothelin-1 (ET-1, vascular endothelial growth factor (VEGF, laminin (LN, and type IV collagen on leptin-stimulated LSECs. We found that normal LSECs had mostly open fenestrae and no organized basement membrane. Leptin-stimulated LSECs showed the formation of a continuous basement membrane with few open fenestrae, which were the features of capillarization. Expression of ET-1, VEGF, LN, and type IV collagen was enhanced in leptin-stimulated LSECs. Plumbagin was used to treat leptin-stimulated LSECs. The sizes and numbers of open fenestrae were markedly decreased, and no basement membrane production was found after plumbagin administration. Plumbagin decreased the levels of ET-1, VEGF, LN, and type IV collagen in leptin-stimulated LSECs. Plumbagin promoted downregulation of ET-1, VEGF, LN, and type IV collagen mRNA. Altogether, our data reveal that plumbagin reverses capillarization of hepatic sinusoids by downregulation of ET-1, VEGF, LN, and type IV collagen.

  4. Distinctive tomographic abnormalities of the craniocervical region in a patient with osteogenesis imperfecta type IV B

    Energy Technology Data Exchange (ETDEWEB)

    Kaissi, Ali Al; Klaushofer, Klaus, E-mail: ali.alkaissi@osteologie.a [Ludwig Boltzmann Institute of Osteology, Vienna (Austria); Grill, Franz [Orthopaedic Hospital of Speising, Vienna (Austria). Paediatric Dept.

    2010-07-01

    Osteogenesis imperfecta is a clinically and genetically heterogeneous group of heritable disorders of connective tissue characterized by reduced bone mass (osteopenia) with associated bone fragility. The resulting skeletal manifestations are due to a generalized deficiency in the development of both membranous and endochondral bone and include markedly thin calvarium with delayed closure of the fontanelles and the sutures and excessive Wormian bone formation. Sillence et al. developed a classification system of OI subtypes: OI type I, which is characterised by blue sclerae; perinatal lethal OI type II, also known as congenital OI; OI type III, a progressively deforming subtype with normal sclera; and OI type IV, which is characterized by a normal sclera. Levin et al. have suggested that OI subtypes could be further divided into type A and B based on the absence or presence of dentinogenesis imperfecta. Basilar impression involves the upward (vertical) migration of the odontoid process into the foramen magnum with a depression in the cranium. Basilar impression is a developmental defect and refers to the infolding of the occipital condyles, an elevation of the clivus, and the posterior cranial fossa of the skull. The soft bones of the skull base allow for progressive infolding of the dysplastic clivus and translocation of the odontoid into the posterior fossa. The combination of platybasia and basilar impression can lead to severe distortion of the spinal cord and the anterior brain stem. The specific structures that can be involved include the upper cervical cord, medulla, pons, mid-brain, cerebellum, as well as the vertebrobasilar system. (author)

  5. [Pavlik harness for the treatment of congenital hip dysplasia types D III and IV].

    Science.gov (United States)

    Zajonz, D; Strobel, S; Wojan, M; von der Höh, N; Brandmaier, P; Josten, C; Schumann, E; Heyde, C-E

    2016-01-01

    Up to 4% of all neonates in Central Europe are born with congenital hip dysplasia (CHD), the most common congenital disease of the musculoskeletal system. However, in this retrospective analysis the outcomes of infants with CHD (type D, III or IV according to Graf) have been considered, with Pavlik therapy starting within the first 12 weeks of life. Connections between the start of therapy or the first finding according to Graf`s classification and the ultrasound result achieved, as well as the X-rays taken after 1 and 2 years, were evaluated. No repositioning under Pavlik treatment or side effects and their relevance have been evaluated, especially with regard to avascular necrosis (AVN) of the femoral head. All infants treated using Pavlik treatment for CHD between 2010 and 2012 in our clinic were determined. A total of 62 patients with 79 pathological hips were included. The infants were classified into three groups to evaluate the influence of the start of therapy on the result: group I with the first investigation and start of treatment within the first 10 days of life, group II between the 11th day and the end of week 3, group III within preventive general examinations (U3) after the 4th week. Clinical examinations and the usual ultrasound scans were performed at an average of 1, 3, and 6 months. Furthermore, after 1 and 2 years clinical and radiological investigations were carried out, as well as further examinations depending on the findings. A failure of repositioning of the Pavlik treatment occurred in group I in 1 case (2.2%), in group II in 1 case (7.1%), and in group III in 2 cases (10%). This occurs in hips type D and type III in 1 case each (3.3%) and type IV in 2 cases (10.5%). Maturation disorders of the hips were found in 1 case (2.2%) in group I, 1 case (7.1%) in group II, and 3 cases (15%) in group III. Avascular necrosis of the femoral head was proven in 2 cases (4.4%) in group I, 0% in group II, and in 1 case (5%) in group III. All patients

  6. AN ASSESSMENT OF THE SERVICE HISTORY AND CORROSION SUSCEPTIBILITY OF TYPE IV WASTE TANKS

    Energy Technology Data Exchange (ETDEWEB)

    Wiersma, B

    2008-09-18

    Type IV waste tanks were designed and built to store waste that does not require auxiliary cooling. Each Type IV tank is a single-shell tank constructed of a steel-lined pre-stressed concrete tank in the form of a vertical cylinder with a concrete domed roof. There are four such tanks in F-area, Tanks 17-20F, and four in H-Area, Tanks 21-24H. Leak sites were discovered in the liners for Tanks 19 and 20F in the 1980's. Although these leaks were visually observed, the investigation to determine the mechanism by which the leaks had occurred was not completed at that time. Therefore, a concern was raised that the same mechanism which caused the leak sites in the Tanks in F-area may also be operable in the H-Area tanks. Data from the construction of the tanks (i.e., certified mill test reports for the steel, no stress-relief), the service history (i.e., waste sample data, temperature data), laboratory tests on actual wastes and simulants (i.e., electrochemical testing), and the results of the visual inspections were reviewed. The following observations and conclusions were made: (1) Comparison of the compositional and microstructural features indicate that the A212 material utilized for construction of the H-Area tanks are far more resistant to SCC than the A285 materials used for construction of the F-Area tanks. (2) A review of the materials of construction, temperature history, service histories concluded that F-Area tanks likely failed by caustic stress corrosion cracking. (3) The environment in the F-Area tanks was more aggressive than that experienced by the H-Area tanks. (4) Based on a review of the service history, the H-Area tanks have not been exposed to an environment that would render the tanks susceptible to either nitrate stress corrosion cracking (i.e., the cause of failures in the Type I and II tanks) or caustic stress corrosion cracking. (5) Due to the very dilute and uninhibited solutions that have been stored in Tank 23H, vapor space corrosion

  7. c-Type cytochrome-dependent formation of U(IV nanoparticles by Shewanella oneidensis.

    Directory of Open Access Journals (Sweden)

    Matthew J Marshall

    2006-09-01

    Full Text Available Modern approaches for bioremediation of radionuclide contaminated environments are based on the ability of microorganisms to effectively catalyze changes in the oxidation states of metals that in turn influence their solubility. Although microbial metal reduction has been identified as an effective means for immobilizing highly-soluble uranium(VI complexes in situ, the biomolecular mechanisms of U(VI reduction are not well understood. Here, we show that c-type cytochromes of a dissimilatory metal-reducing bacterium, Shewanella oneidensis MR-1, are essential for the reduction of U(VI and formation of extracellular UO(2 nanoparticles. In particular, the outer membrane (OM decaheme cytochrome MtrC (metal reduction, previously implicated in Mn(IV and Fe(III reduction, directly transferred electrons to U(VI. Additionally, deletions of mtrC and/or omcA significantly affected the in vivo U(VI reduction rate relative to wild-type MR-1. Similar to the wild-type, the mutants accumulated UO(2 nanoparticles extracellularly to high densities in association with an extracellular polymeric substance (EPS. In wild-type cells, this UO(2-EPS matrix exhibited glycocalyx-like properties and contained multiple elements of the OM, polysaccharide, and heme-containing proteins. Using a novel combination of methods including synchrotron-based X-ray fluorescence microscopy and high-resolution immune-electron microscopy, we demonstrate a close association of the extracellular UO(2 nanoparticles with MtrC and OmcA (outer membrane cytochrome. This is the first study to our knowledge to directly localize the OM-associated cytochromes with EPS, which contains biogenic UO(2 nanoparticles. In the environment, such association of UO(2 nanoparticles with biopolymers may exert a strong influence on subsequent behavior including susceptibility to oxidation by O(2 or transport in soils and sediments.

  8. Altered stress fibers and integrin expression in the Malpighian epithelium of Drosophila type IV collagen mutants

    Directory of Open Access Journals (Sweden)

    András A. Kiss

    2016-06-01

    Full Text Available Basement membranes (BMs are highly specialized extracellular matrices (ECMs that provide support and polarization cues for epithelial cells. Proper adhesion to the BM is pivotal in epithelial cell function and survival. Type IV collagens are the predominant components of all types of BMs, that form an irregular, polygonal lattice and serve as a scaffold for numerous other BM components and BM-associated cells. Mutations in the ubiquitous human BM components COL4A1 and COL4A2 cause a multisystem disorder involving nephropathy. Affected patients develop renal dysfunction and chronic kidney failure with or without hematuria. Mouse Col4a1 and Col4a2 mutants recapitulate the human symptoms. In vertebrates, excretion is accomplished by the kidneys and by the Malpighian tubules in insects, including the fruit fly Drosophila. Our present results with dominant, temperature-sensitive mutation of the Drosophila col4a1 gene demonstrate altered integrin expression and amplified effects of mechanical stress on the Malpighian epithelial cytoskeleton.

  9. Xanthomonas citri subsp. citri type IV Pilus is required for twitching motility, biofilm development, and adherence.

    Science.gov (United States)

    Dunger, German; Guzzo, Cristiane R; Andrade, Maxuel O; Jones, Jeffrey B; Farah, Chuck S

    2014-10-01

    Bacterial type IV pili (T4P) are long, flexible surface filaments that consist of helical polymers of mostly pilin subunits. Cycles of polymerization, attachment, and depolymerization mediate several pilus-dependent bacterial behaviors, including twitching motility, surface adhesion, pathogenicity, natural transformation, escape from immune system defense mechanisms, and biofilm formation. The Xanthomonas citri subsp. citri strain 306 genome codes for a large set of genes involved in T4P biogenesis and regulation and includes several pilin homologs. We show that X. citri subsp. citri can exhibit twitching motility in a manner similar to that observed in other bacteria such as Pseudomonas aeruginosa and Xylella fastidiosa and that this motility is abolished in Xanthomonas citri subsp. citri knockout strains in the genes coding for the major pilin subunit PilAXAC3241, the ATPases PilBXAC3239 and PilTXAC2924, and the T4P biogenesis regulators PilZXAC1133 and FimXXAC2398. Microscopy analyses were performed to compare patterns of bacterial migration in the wild-type and knockout strains and we observed that the formation of mushroom-like structures in X. citri subsp. citri biofilm requires a functional T4P. Finally, infection of X. citri subsp. citri cells by the bacteriophage (ΦXacm4-11 is T4P dependent. The results of this study improve our understanding of how T4P influence Xanthomonas motility, biofilm formation, and susceptibility to phage infection.

  10. Type IV Hypersensitivity to Gold Weight Upper-Eyelid Implant: Case Report and Review of the Literature.

    Science.gov (United States)

    Kilduff, Caroline L S; Casswell, Edward J; Imonikhe, Richard; Marjanovic, Branka

    2017-05-04

    Complications associated with gold-weight insertion for lagophthalmos are uncommon, recent reports have provided evidence to suggest that type IV hypersensitivity to gold can cause a persistent inflammatory reaction. We present a case of a 46-year-old man who experienced persistent post-operative inflammation, and summarize previously documented cases. This patient underwent uncomplicated insertion of an upper eyelid gold weight for right-sided facial nerve palsy. He had no allergies or implanted metalwork. Post-operatively erythema was noted at seven-weeks and did not resolve. The weight was removed after six-months. The histopathological findings were in keeping with type IV hypersensitivity and similar to previous cases. Although infrequent, this complication has poor outcomes. The definitive management is removal of the weight. Information regarding implanted gold, and previous reactions should be elicited pre-operatively. Type IV hypersensitivity should be considered in patients with persistent inflammation that do not respond to antibiotic or steroid therapy.

  11. Delayed type IV hypersensitivity reaction to porcine acellular dermal matrix masquerading as infection resulting in multiple debridements.

    Science.gov (United States)

    Vedak, P; St John, J; Watson, A; Garibyan, L; Mihm, M C; Nazarian, R M; Levins, P C; Cetrulo, C L; Schalock, P; Kroshinsky, D

    2017-06-01

    Delayed type IV hypersensitivity reactions are well established in the surgical setting with respect to external exposure via topical antibiotics and internal exposure via synthetic materials. In contrast, biologic matrix is derived from decellularized human or animal tissues and is consequently believed to elicit a minimal host inflammatory response. We report a case of delayed type IV hypersensitivity reaction secondary to a biologic comprised of porcine-derived acellular dermal matrix, [Strattice™]. While biologic matrix is often preferred over synthetic mesh due to its decreased risk for infection, this case emphasizes that potential for hypersensitivity to the material persists. Type IV hypersensitivity reactions should be included in the differential diagnosis for suspected post-operative infections.

  12. Genetic analysis of a Chinese family with members affected with Usher syndrome type II and Waardenburg syndrome type IV.

    Science.gov (United States)

    Wang, Xueling; Lin, Xiao-Jiang; Tang, Xiangrong; Chai, Yong-Chuan; Yu, De-Hong; Chen, Dong-Ye; Wu, Hao

    2017-11-01

    The purpose of this study was to identify the genetic causes of a family presenting with multiple symptoms overlapping Usher syndrome type II (USH2) and Waardenburg syndrome type IV (WS4). Targeted next-generation sequencing including the exon and flanking intron sequences of 79 deafness genes was performed on the proband. Co-segregation of the disease phenotype and the detected variants were confirmed in all family members by PCR amplification and Sanger sequencing. The affected members of this family had two different recessive disorders, USH2 and WS4. By targeted next-generation sequencing, we identified that USH2 was caused by a novel missense mutation, p.V4907D in GPR98; whereas WS4 due to p.V185M in EDNRB. This is the first report of homozygous p.V185M mutation in EDNRB in patient with WS4. This study reported a Chinese family with multiple independent and overlapping phenotypes. In condition, molecular level analysis was efficient to identify the causative variant p.V4907D in GPR98 and p.V185M in EDNRB, also was helpful to confirm the clinical diagnosis of USH2 and WS4. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Thickness and multi-axial stress creep rupture criteria of the Type IV component of a ferritic steel weld

    OpenAIRE

    Hayhurst, R James; Vakili-Tahami, Farid; Mustata, Radu; Hayhurst, David

    2004-01-01

    The paper summarises previously derived constitutive parameters for the temperatures 575, 590, 600, 620 and 640oC. Values of the multi-axial stress rupture parameter are reviewed and recorded. This constitutive parameter set is used to determine the thickness of the Type IV material zone to be 0.7mm. Next, values of Type IV multi-axial stress rupture parameter are determined for a wide range of butt-welded pipe and crosswelded uni-axial specimens, and an interpolation equation has been derive...

  14. Human corneal basement membrane heterogeneity: topographical differences in the expression of type IV collagen and laminin isoforms.

    Science.gov (United States)

    Ljubimov, A V; Burgeson, R E; Butkowski, R J; Michael, A F; Sun, T T; Kenney, M C

    1995-04-01

    The corneal epithelium converges at the peripheral zone (limbus) with the conjunctival epithelium, forming a continuous sheet with phenotypically distinct regions--central, limbal, and conjunctival. The epithelial basement membrane (EBM) is important for corneal functions and cell adhesion, but its regional composition is poorly understood. Current literature is controversial as to the occurrence of type IV collagen in the cornea. The aim of this study was to investigate in detail corneal basement membrane (BM) composition and correlate it with the differentiation state of contributing cells. Adult human corneas (N = 8) were cryosectioned and analyzed by immunofluorescence with antibodies to 15 BM components and to keratin 3, a marker of corneal epithelial differentiation. A novel type of spatial heterogeneity ("horizontal") in the EBM composition was found between the central cornea, limbus, and conjunctiva. Central EBM had type IV collagen alpha 3-alpha 5 chains, whereas limbal and conjunctival EBM contained alpha 1-alpha 2 chains and also laminin alpha 2 and beta 2 chains. Limbal EBM in addition had alpha 5(IV) chain. Laminin-1 (alpha 1 beta 1 gamma 1), laminin-5 (alpha 3 beta 3 gamma 2), perlecan, fibronectin, entactin/nidogen, and type VII collagen were seen in the entire EBM. Another novel type of BM heterogeneity ("vertical") was typical for the corneal Descemet's membrane: its stromal face had alpha 1(IV) and alpha 2(IV) chains and fibronectin, whereas alpha 3(IV)-alpha 5(IV) chains, entactin/nidogen, laminin-1, and perlecan were present on the endothelial face. Type IV collagen controversy is the result of the shifts of isoforms in the limbus and conjunctiva. These shifts and the appearance of additional laminins in the limbus may be related to the differentiation state of corneal cells contributing to the EBM formation. Novel types of BM heterogeneity in the human cornea are described: regional (horizontal) in the EBM and vertical in the Descemet

  15. Stabilization technique for columella using trimmed autologous temporal fascia in type III and IV tympanoplasty--Muffler method.

    Science.gov (United States)

    Kanemaru, Shin-Ichi; Ito, Juichi; Tsuji, Jun; Fujino, Kiyohiro; Hiraumi, Harukazu; Omori, Koichi

    2007-02-01

    High success rates of recovery of hearing level in type III and IV tympanoplasty could be achieved by this stabilization technique for columella using trimmed autologous temporal fascia. The aim of this study was to evaluate a new stabilization technique for columella using trimmed autologous temporal fascia in type III and IV tympanoplasty. A total of 55 patients (21 male, 34 female, aged 4-85 years) with chronic otitis media (n=16) and cholesteatoma (n=39) underwent tympanoplasty using this new stabilization technique for columella. Thirty-one patients underwent type III tympanoplasty and 24 patients underwent type IV tympanoplasty. Forty-two patients underwent a staged operation and 13 patients underwent a single operation. The observation period was 3.5 years from 6 months after the last operation. The overall success rates in type III and IV tympanoplasty were 87.1% (27/31) and 83.3% (20/24), respectively. Two of eight patients for whom the procedure was unsuccessful underwent reoperation and they acquired good hearing.

  16. Protein distribution and gene expression of collagen type IV in the neonatal rat ovary during follicle formation.

    Science.gov (United States)

    Rajah, R; Sundaram, G S

    1994-09-01

    Protein distribution and mRNA expression of basement membrane collagen (type IV) during follicle formation were studied using serial sections from 24, 48 and 72 hrs. old rat ovaries. Collagen type IV, a protein found only in the basal lamina of the basement membrane, was localized under light microscope using a polyclonal antibody. During the first 24 hrs. postpartum, immunostaining was found as thin septa separating the clusters from the stroma. By 72 hrs. postpartum, immunostaining was found around each newly formed primordial follicle. The cell types involved in collagen type IV synthesis were determined by in situ hybridization using a biotinylated riboprobe. Before the follicles had been formed, the stromal cells showed intense staining while the epithelial presumptive granulosa cells showed a pale staining. However, after a follicle had been formed, some of the granulosa cells enclosed within the follicular basement membrane showed strong staining for the message. The presumptive granulosa cells are presumed to be the progenitors of granulosa cells. If so, these observations suggest that the expression of the message coding for collagen type IV by the granulosa cells may be a marker for commitment of the undifferentiated cell to the granulosa cell lineage.

  17. Pseudoaneurysm Accompanied by Crowe Type IV Developmental Dysplasia of the Hip: A Case Report

    Directory of Open Access Journals (Sweden)

    Hirotake Yo

    2012-01-01

    Full Text Available We report the case of a 72-year-old woman whose pseudoaneurysm was difficult to diagnose and treat. The patient had a history of congenital dislocated hip and was undergoing anticoagulation therapy with warfarin due to the mitral valve replacement. Her chief complaint was pain and enlargement of the left buttock, and the laboratory tests revealed severe anemia. However, her elderly depression confused her chief complaint, and she was transferred to a psychiatric hospital. Two months after the onset of the symptoms, she was finally diagnosed with a pseudoaneurysm by contrast-enhanced CT and angiography. IDC coils were used for embolization. A plain CT showed hemostasis as well as a reduced hematoma at 2 months after the embolization. The possible contributing factors for the pseudoaneurysm included bleeding due to warfarin combined with an intramuscular hematoma accompanied by Crowe type IV developmental dysplasia of the hip that led to an arterial rupture by impingement between pelvis and femoral head. Since the warfarin treatment could not be halted due to the valve replacement, embolization was chosen for her treatment, and the treatment outcome was favorable.

  18. Structure of the outer membrane complex of a type IV secretion system

    Science.gov (United States)

    Chandran, Vidya; Fronzes, Rémi; Duquerroy, Stéphane; Cronin, Nora; Navaza, Jorge; Waksman, Gabriel

    2009-01-01

    Type IV secretion systems are secretion nanomachines spanning the two membranes of Gram-negative bacteria. Three proteins, VirB7, VirB9, and VirB10 assemble into a 1.05 MDa core spanning the inner and outer membranes. This core consists of 14 copies of each of the proteins and forms two layers, the I and O layers, inserting in the inner and outer membrane, respectively. Here we present the crystal structure of a ~0.6 MDa outer membrane complex containing the entire O-layer. This structure is the largest determined for an outer membrane channel and is also unprecedented in being composed of three proteins. Unexpectedly, this structure identifies VirB10 as the outer membrane channel with a unique hydrophobic double helical trans-membrane region. This structure establishes VirB10 as the only known protein crossing both membranes of Gram-negative bacteria. Comparison of the cryo-EM and crystallographic structures point to conformational changes regulating channel opening and closing. PMID:19946264

  19. Type IV pilus proteins form an integrated structure extending from the cytoplasm to the outer membrane.

    Directory of Open Access Journals (Sweden)

    Chengyun Li

    Full Text Available The bacterial type IV pilus (T4P is the strongest biological motor known to date as its retraction can generate forces well over 100 pN. Myxococcus xanthus, a δ-proteobacterium, provides a good model for T4P investigations because its social (S gliding motility is powered by T4P. In this study, the interactions among M. xanthus T4P proteins were investigated using genetics and the yeast two-hybrid (Y2H system. Our genetic analysis suggests that there is an integrated T4P structure that crosses the inner membrane (IM, periplasm and the outer membrane (OM. Moreover, this structure exists in the absence of the pilus filament. A systematic Y2H survey provided evidence for direct interactions among IM and OM proteins exposed to the periplasm. For example, the IM lipoprotein PilP interacted with its cognate OM protein PilQ. In addition, interactions among T4P proteins from the thermophile Thermus thermophilus were investigated by Y2H. The results indicated similar protein-protein interactions in the T4P system of this non-proteobacterium despite significant sequence divergence between T4P proteins in T. thermophilus and M. xanthus. The observations here support the model of an integrated T4P structure in the absence of a pilus in diverse bacterial species.

  20. Type IV traffic ATPase TrwD as molecular target to inhibit bacterial conjugation.

    Science.gov (United States)

    Ripoll-Rozada, Jorge; García-Cazorla, Yolanda; Getino, María; Machón, Cristina; Sanabria-Ríos, David; de la Cruz, Fernando; Cabezón, Elena; Arechaga, Ignacio

    2016-06-01

    Bacterial conjugation is the main mechanism responsible for the dissemination of antibiotic resistance genes. Hence, the search for specific conjugation inhibitors is paramount in the fight against the spread of these genes. In this pursuit, unsaturated fatty acids have been found to specifically inhibit bacterial conjugation. Despite the growing interest on these compounds, their mode of action and their specific target remain unknown. Here, we identified TrwD, a Type IV secretion traffic ATPase, as the molecular target for fatty acid-mediated inhibition of conjugation. Moreover, 2-alkynoic fatty acids, which are also potent inhibitors of bacterial conjugation, are also powerful inhibitors of the ATPase activity of TrwD. Characterization of the kinetic parameters of ATPase inhibition has led us to identify the catalytic mechanism by which fatty acids exert their activity. These results open a new avenue for the rational design of inhibitors of bacterial conjugation in the fight against the dissemination of antibiotic resistance genes. © 2016 John Wiley & Sons Ltd.

  1. Structure of a translocation signal domain mediating conjugative transfer by type IV secretion systems.

    Science.gov (United States)

    Redzej, Adam; Ilangovan, Aravindan; Lang, Silvia; Gruber, Christian J; Topf, Maya; Zangger, Klaus; Zechner, Ellen L; Waksman, Gabriel

    2013-07-01

    Relaxases are proteins responsible for the transfer of plasmid and chromosomal DNA from one bacterium to another during conjugation. They covalently react with a specific phosphodiester bond within DNA origin of transfer sequences, forming a nucleo-protein complex which is subsequently recruited for transport by a plasmid-encoded type IV secretion system. In previous work we identified the targeting translocation signals presented by the conjugative relaxase TraI of plasmid R1. Here we report the structure of TraI translocation signal TSA. In contrast to known translocation signals we show that TSA is an independent folding unit and thus forms a bona fide structural domain. This domain can be further divided into three subdomains with striking structural homology with helicase subdomains of the SF1B family. We also show that TSA is part of a larger vestigial helicase domain which has lost its helicase activity but not its single-stranded DNA binding capability. Finally, we further delineate the binding site responsible for translocation activity of TSA by targeting single residues for mutations. Overall, this study provides the first evidence that translocation signals can be part of larger structural scaffolds, overlapping with translocation-independent activities. © 2013 The Authors. Molecular Microbiology published by John Wiley & Sons Ltd.

  2. Type IV pili of Acidithiobacillus ferrooxidans can transfer electrons from extracellular electron donors.

    Science.gov (United States)

    Li, Yongquan; Li, Hongyu

    2014-03-01

    Studies on Acidithiobacillus ferrooxidans accepting electrons from Fe(II) have previously focused on cytochrome c. However, we have discovered that, besides cytochrome c, type IV pili (Tfp) can transfer electrons. Here, we report conduction by Tfp of A. ferrooxidans analyzed with a conducting-probe atomic force microscope (AFM). The results indicate that the Tfp of A. ferrooxidans are highly conductive. The genome sequence of A. ferrooxidans ATCC 23270 contains two genes, pilV and pilW, which code for pilin domain proteins with the conserved amino acids characteristic of Tfp. Multiple alignment analysis of the PilV and PilW (pilin) proteins indicated that pilV is the adhesin gene while pilW codes for the major protein element of Tfp. The likely function of Tfp is to complete the circuit between the cell surface and Fe(II) oxides. These results indicate that Tfp of A. ferrooxidans might serve as biological nanowires transferring electrons from the surface of Fe(II) oxides to the cell surface. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Polarization Characteristics of Zebra Patterns in Type IV Solar Radio Bursts

    Science.gov (United States)

    Kaneda, K.; Misawa, H.; Iwai, K.; Tsuchiya, F.; Obara, T.; Katoh, Y.; Masuda, S.

    2017-06-01

    The polarization characteristics of zebra patterns (ZPs) in type IV solar bursts were studied. We analyzed 21 ZP events observed by the Assembly of Metric-band Aperture Telescope and Real-time Analysis System between 2010 and 2015 and identified the following characteristics: a degree of circular polarization (DCP) in the range of 0%-70%, a temporal delay of 0-70 ms between the two circularly polarized components (i.e., the right- and left-handed components), and dominant ordinary-mode emission in about 81% of the events. For most events, the relation between the dominant and delayed components could be interpreted in the framework of fundamental plasma emission and depolarization during propagation, though the values of DCP and delay were distributed across wide ranges. Furthermore, it was found that the DCP and delay were positively correlated (rank correlation coefficient R = 0.62). As a possible interpretation of this relationship, we considered a model based on depolarization due to reflections at sharp density boundaries assuming fundamental plasma emission. The model calculations of depolarization including multiple reflections and group delay during propagation in the inhomogeneous corona showed that the DCP and delay decreased as the number of reflections increased, which is consistent with the observational results. The dispersive polarization characteristics could be explained by the different numbers of reflections causing depolarization.

  4. Proximal spinal muscular atrophy types I-IV: Specific features of molecular genetic diagnosis

    Directory of Open Access Journals (Sweden)

    V. V. Zabnenkova

    2013-01-01

    Full Text Available Proximal spinal muscular atrophy (SMA types I-IV is the most common autosomal recessive neuromuscular disease caused by mutations in the SMN1 gene encoding the survival motor neuron protein. It is characterized by progressive muscle weakness due to injury of the motor neurons of the anterior horns of the spinal cord. The classification of the disease is based on the time of its onset, severity, and survival. The detection of the major mutation of exon 7 and/or 8 deletion in the SMN1 gene is a qualitative reliable and sensitive diagnostic test. The SMN1 gene has the almost complete homolog SMN2 gene, which hampers the analysis of heterozygous carriage of the disease. So the determination of the carriage status is based on the quantitative analysis of the number of SMN1 gene copies. The paper covers problems and new possibilities in themolecular genetic diagnosis of proximal SMA.

  5. Inhibition of inflammasome activation by Coxiella burnetii type IV secretion system effector IcaA

    Science.gov (United States)

    Cunha, Larissa D.; Ribeiro, Juliana M.; Fernandes, Talita D.; Massis, Liliana M.; Khoo, Chen Ai; Moffatt, Jennifer H.; Newton, Hayley J.; Roy, Craig R.; Zamboni, Dario S.

    2015-01-01

    Coxiella burnetii is a highly infectious bacterium that promotes its own replication in macrophages by inhibiting several host cell responses. Here, we show that C. burnetii inhibits caspase-1 activation in primary mouse macrophages. By using co-infection experiments, we determine that the infection of macrophages with C. burnetii inhibits the caspase-11-mediated non-canonical activation of the NLRP3 inflammasome induced by subsequent infection with Escherichia coli or Legionella pneumophila. Genetic screening using flagellin mutants of L. pneumophila as a surrogate host, reveals a novel C. burnetii gene (IcaA) involved in the inhibition of caspase activation. Expression of IcaA in L. pneumophila inhibited the caspase-11 activation in macrophages. Moreover, icaA- mutants of C. burnetii failed to suppress the caspase-11-mediated inflammasome activation induced by L. pneumophila. Our data reveal IcaA as a novel C. burnetii effector protein that is secreted by the Dot/Icm type IV secretion system and interferes with the caspase-11-induced, non-canonical activation of the inflammasome. PMID:26687278

  6. Laparoscopic anterior gastropexy for type III/IV hiatal hernia in elderly patients.

    Science.gov (United States)

    Higashi, Shigeyoshi; Nakajima, Kiyokazu; Tanaka, Koji; Miyazaki, Yasuhiro; Makino, Tomoki; Takahashi, Tsuyoshi; Kurokawa, Yukinori; Yamasaki, Makoto; Takiguchi, Shuji; Mori, Masaki; Doki, Yuichiro

    2017-12-01

    Large esophageal hiatal hernias occur most commonly in elderly patients with comorbidities, in whom even an elective surgery cannot be performed without high risks. Although fundoplication is recommended for esophageal hiatal hernia repair, we prefer not to limit our options to fundoplication, as obstruction is a frequent main complaint. We favor an anterior gastropexy approach instead to perform anti-reflux surgery and prevent recurrent protrusion and torsion of the incarcerated organ with minimal risk. The aim was to evaluate the safety and effectiveness of anterior gastropexy for large hiatal hernia in elderly patients with comorbidities. We retrospectively evaluated 8 patients who underwent laparoscopic anterior gastropexy for large hiatal hernia (type III or IV) since 2006. All patients were women with a median age of 82 years (range, 74-87 years). The major complaint was obstruction in all patients, with relatively mild reflux symptoms. They underwent successful laparoscopic surgery with no conversion to laparotomy. Fundoplication was performed in 4 cases. No perioperative complications occurred, and the main complaint resumed rapidly in all patients, without recurrence during postoperative follow-up of median 48 months (range, 5-77 months). Laparoscopic anterior gastropexy is safe and effective and can be considered as one of the practical surgical options for large hiatal hernias in elderly patients, whom surgical intervention should be minimized due to their comorbidities.

  7. Immunogold fine structural localization of extracellular matrix components in aged human cornea. I. Types I-IV collagen and laminin.

    Science.gov (United States)

    Marshall, G E; Konstas, A G; Lee, W R

    1991-01-01

    Using the immunogold technique combined with cryoultramicrotomy and London Resin white (LR white) embedding, we studied the fine structural distribution of types I-IV collagen and laminin in corneal tissue from seven enucleated human eyes (age range, 63-78 years). Type II collagen was not identified in any corneal layer. Type I and type III collagen were distributed in a similar fashion in striated collagen fibrils in Bowman's layer and in the stroma. Type IV collagen was located only in the posterior non-banded region of Descemet's membrane. Laminin was identified in subepithelial anchoring plaques and the sub-endothelial region of Descemet's membrane in accordance with its recognized adhesive function.

  8. A novel basolateral type IV secretion model for the CagA oncoprotein of Helicobacter pylori

    Directory of Open Access Journals (Sweden)

    Silja Wessler

    2017-12-01

    Full Text Available Intercellular junctions are crucial structural elements for the formation and maintenance of epithelial barrier functions to control homeostasis or protect against intruding pathogens in humans. Alterations in these complexes represent key events in the development and progression of numerous cancers as well as multiple infectious diseases. Many bacterial pathogens harbor type IV secretion systems (T4SSs, which translocate virulence factors into host cells to hijack cellular processes. The pathology of the gastric pathogen and type-I carcinogen Helicobacter pylori strongly depends on a T4SS encoded by the cag pathogenicity island (cagPAI. This T4SS forms a needle-like pilus and its activity is accomplished by the pilus-associated factors CagL, CagI and CagY which target the host integrin-β1 receptor followed by injection of the CagA oncoprotein into non-polarized AGS gastric epithelial cells. The finding of a T4SS receptor, however, suggested the presence of a sophisticated control mechanism for the injection of CagA. In fact, integrins constitute a group of basolateral receptors, which are normally absent at apical surfaces of the polarized epithelium in vivo. Our new results demonstrate that T4SS-pilus formation during H. pylori infection of polarized epithelial cells occurs preferentially at basolateral sites, and not at apical membranes (Tegtmeyer et al., 2017. We propose a stepwise process how H. pylori interacts with components of intercellular tight junctions (TJs and adherens junctions (AJs, followed by contacting integrin-based focal adhesions to disrupt and transform the epithelial cell layer in the human stomach. The possible impact of this novel signaling cascade on pathogenesis during infection is reviewed.

  9. Topical prostaglandin F2alpha treatment reduces collagen types I, III, and IV in the monkey uveoscleral outflow pathway.

    Science.gov (United States)

    Sagara, T; Gaton, D D; Lindsey, J D; Gabelt, B T; Kaufman, P L; Weinreb, R N

    1999-06-01

    Topical prostaglandin F2alpha isopropyl ester increases uveoscleral outflow in monkeys and humans. To investigate the effects of prostaglandin F2alpha isopropyl ester with topical administration on collagen types I, III, and IV within the anterior segment tissue of monkey eyes. Eight eyes of 4 cynomolgus monkeys were evaluated. One eye of each monkey was treated with 2 microg of prostaglandin F2alpha isopropyl ester twice daily for 5 days, and intraocular pressure reduction was confirmed. These eyes were fixed in methacarn, and paraffin sections were immunostained using antibodies to collagen types I, II, or IV. To measure staining intensity, optical density (OD) was determined using 2-dimensional imaging densitometry. Mean OD scores along line segments placed over the ciliary muscle were determined. Mean+/-SD OD scores for collagen types I, III, and IV were less in the ciliary muscle of prostaglandin-treated eyes than in vehicle-treated eyes by 52%+/-7%, 45%+/-6%, and 45%+/-5%, respectively. In the sclera adjacent to the ciliary body, mean OD scores for collagen types I and III were less in prostaglandin-treated eyes, by 43%+/-32% and 45%+/-13%, respectively. The scleral stroma was minimally immunoreactive for collagen type IV. All differences were significant by the paired Student t test (Pcollagen types I, III, and IV immunoreactivity in the ciliary muscle and adjacent sclera following topical prostaglandin F2alpha isopropyl ester treatment. These reductions may contribute to the increased uveoscleral outflow observed with topical prostaglandin treatment. The cellular mechanism by which certain prostaglandins lower intraocular pressure is not known. The present study provides immunohistochemical data demonstrating that intraocular pressure reduction that occurs with topical prostaglandin F2alpha is associated with a reduction of collagens within the uveoscleral outflow pathway.

  10. Ehlers-Danlos syndrome type IV is associated with a novel G984R COL3A1 mutation.

    Science.gov (United States)

    Deng, Yao; Wei, Shijie; Hu, Shijun; Chen, Jinlan; Tan, Zhiping; Yang, Yifeng

    2015-07-01

    Ehlers-Danlos syndrome type IV is an autosomal dominant connective tissue disease. Mutations in COL3A1 have been identified to underlie this disease; however, to the best of our knowledge, no COL3A1 mutations have been reported in Ehlers-Danlos syndrome type IV patients with an ascending aortic aneurysm. In order to develop further understanding of COL3A1 mutations, an Ehlers-Danlos syndrome type IV patient diagnosed with an ascending aortic aneurysm and a familial history of sudden mortality was analyzed. Genomic DNA was isolated from the peripheral blood of the patient and his family members. All coding exons of eight aneurysm-related genes (FBN1, TGFBR1, TGFBR 2, MYH11, ACTA2, SLC2A10, NOTCH1 and COL3A1) were amplified using polymerase chain reaction (PCR). The PCR products were sequenced with the ABI 3100 Genetic Analyzer, and a mutation was predicted and identified using Polyphen-2, SIFT and Mutation Taster. The novel mutation was identified as c.2950G>A in COL3A1, which results in p.G984R. All three programs predicted this mutation to be deleterous to the protein function. The novel mutation identified in this study is potentially responsible for Ehlers-Danlos syndrome type IV in this patient, and expands the spectrum of COL3A1 mutations.

  11. A novel relaxase homologue is involved in chromosomal DNA processing for type IV secretion in Neisseria gonorrhoeae

    NARCIS (Netherlands)

    Salgado-Pabon, Wilmara; Jain, Samta; Turner, Nicholas; van der Does, Chris; Dillard, Joseph P.

    2007-01-01

    The Neisseria gonorrhoeae type IV secretion system secretes chromosomal DNA that acts in natural transformation. To examine the mechanism of DNA processing for secretion, we made mutations in the putative relaxase gene traI and used nucleases to characterize the secreted DNA. The nuclease

  12. The specific type IV phosphodiesterase inhibitor rolipram differentially regulates the proinflammatory mediators TNF-alpha and nitric oxide

    NARCIS (Netherlands)

    Greten, T F; Eigler, A; Sinha, B; Moeller, J; Endres, S

    We compared the effect of the specific type IV phosphodiesterase inhibitor rolipram on intracellular cAMP concentration, nitric oxide (NO) and tumour necrosis factor-alpha (TNF) formation in the murine macrophage cell line RAW 264.7. We found a dose-dependent increase of nitrite accumulation in

  13. BIOLOGY OF BACTERIUM LEPISEPTICUM : IV. VIRULENCE OF DIFFUSE AND MUCOID TYPES AND THEIR VARIANTS.

    Science.gov (United States)

    Webster, L T; Burn, C G

    1926-08-31

    bipolar granules and certain quite definite fermentative reactions is placed in this group and is further designated by prefixing the name of the animal from which it was obtained. Differentiation on the basis of serological studies has been extremely difficult and as yet not sufficiently detailed and complete to be of use. We have, therefore, called our strains "Bact. lepisepticum," because they belong to the Pasteurella group and were obtained from rabbits, but at the same time have recognized the possibility that similar forms may also be present in other animal hosts. The "D" and mucoid types may or may not be mutually related. Each has its distinctive growth characteristics and antigenic properties, but these differences are not sufficient to exclude the possibility that the two types may have resulted from the action of bacteriophage or so called "mutation" phenomena upon a single "parent" strain. We have attempted to explain the difference in natural prevalence of "D" and mucoid types in terms of virulence and vegetative capacity (1, a, d). Thus we find that "D" types are at present rarely encountered in rabbits, while the mucoid forms are widespread. The former prove to be highly virulent and slightly vegetative; the latter, less virulent and readily vegetative. The same inverse relationship between virulence and prevalence obtains in human diphtheria (7) and pneumococcus infections (8);-virulent Type I and II pneurnococci and diphtheria bacteria are relatively uncommon in carriers, while the less pathogenic diphtheria forms and Type III and IV pneumococci are widespread. It is possible, therefore, that in these respiratory diseases the virulent types of bacteria are transients, unable to survive in a community except at epidemic times, while the less pathogenic types are more adapted to a parasitic existence in the tissues of a considerable percentage of a population (9). Just what properties of bacteria are responsible for their ability to kill or to survive

  14. Streptocollin, a type IV lanthipeptide produced by Streptomyces collinus Tü 365

    DEFF Research Database (Denmark)

    Iftime, Dumitrita; Jasyk, Martin; Kulik, Andreas

    2015-01-01

    Lanthipeptides are ribosomally synthesized and posttranslationally modified microbial secondary metabolites. Here, we report the identification and isolation of streptocollin from Streptomyces collinus Tü 365, a new member of the class IV lanthipeptides. Insertion of the constitutive ermE* promoter...

  15. [Reactivity of antibodies to collagen types I to IV and antibodies to chondroitin sulfate in the spleen].

    Science.gov (United States)

    Galbavý, S; Ruzicková, M; Surmíková, E; Danihel, L; Porubský, J; Papincák, J; Holesa, S; Trnka, J

    1996-02-01

    Antibodies to collagen type I and III reacted negatively, antibodies to collagen type IV positively with reticulin, trabeculae and circumferent reticulum of lymphatic sheaths, poorly positively with capsula, strongly positively with subcapsular zone. Antibodies to collagen type II reacted positively with capsula, poorly with subcapsular zone, strongly with sinus wall and poorly with trabeculae. They did not react with circumferent reticulum of periarterial lymphoid sheaths. Antibodies to collagen type II and IV reacted positively with central arteries. Antibodies to chondroitinsulphate C reacted poorly and antibodies to chondroitinsulphate B strongly positively with sinus walls and oval cells spread in the white and red pulpa. Antibodies to chondroitin sulphate A reacted similarly as antibodies to chondroitinsulphate B.

  16. Effect of Dermal Thickness on Scars in Women with Type III-IV Fitzpatrick Skin.

    Science.gov (United States)

    Ince, Bilsev; Dadaci, Mehmet; Oltulu, Pembe; Altuntas, Zeynep; Bilgen, Fatma

    2015-06-01

    type was Fitzpatrick type IV (33 patients). The average total scar score was 14 (range 8-25). The total scar score was not significantly associated with dermal thickness. The scar width in patients with a dermal thickness of thickness of ≥0.5 cm. Scar vascularity and noticeability were observed less often in patients with Fitzpatrick skin type III, regardless of scar width. The study findings show that increased dermal thickness is a risk factor for wide scar formation. This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

  17. Functional analysis of an unusual type IV pilus in the Gram‐positive Streptococcus sanguinis

    Science.gov (United States)

    Gurung, Ishwori; Spielman, Ingrid; Davies, Mark R.; Lala, Rajan; Gaustad, Peter; Biais, Nicolas

    2015-01-01

    Summary Type IV pili (Tfp), which have been studied extensively in a few Gram‐negative species, are the paradigm of a group of widespread and functionally versatile nano‐machines. Here, we performed the most detailed molecular characterisation of Tfp in a Gram‐positive bacterium. We demonstrate that the naturally competent S treptococcus sanguinis produces retractable Tfp, which like their Gram‐negative counterparts can generate hundreds of piconewton of tensile force and promote intense surface‐associated motility. Tfp power ‘train‐like’ directional motion parallel to the long axis of chains of cells, leading to spreading zones around bacteria grown on plates. However, S . sanguinis  Tfp are not involved in DNA uptake, which is mediated by a related but distinct nano‐machine, and are unusual because they are composed of two pilins in comparable amounts, rather than one as normally seen. Whole genome sequencing identified a locus encoding all the genes involved in Tfp biology in S . sanguinis. A systematic mutational analysis revealed that Tfp biogenesis in S . sanguinis relies on a more basic machinery (only 10 components) than in Gram‐negative species and that a small subset of four proteins dispensable for pilus biogenesis are essential for motility. Intriguingly, one of the piliated mutants that does not exhibit spreading retains microscopic motility but moves sideways, which suggests that the corresponding protein controls motion directionality. Besides establishing S . sanguinis as a useful new model for studying Tfp biology, these findings have important implications for our understanding of these widespread filamentous nano‐machines. PMID:26435398

  18. Functional analysis of an unusual type IV pilus in the Gram-positive Streptococcus sanguinis.

    Science.gov (United States)

    Gurung, Ishwori; Spielman, Ingrid; Davies, Mark R; Lala, Rajan; Gaustad, Peter; Biais, Nicolas; Pelicic, Vladimir

    2016-01-01

    Type IV pili (Tfp), which have been studied extensively in a few Gram-negative species, are the paradigm of a group of widespread and functionally versatile nano-machines. Here, we performed the most detailed molecular characterisation of Tfp in a Gram-positive bacterium. We demonstrate that the naturally competent Streptococcus sanguinis produces retractable Tfp, which like their Gram-negative counterparts can generate hundreds of piconewton of tensile force and promote intense surface-associated motility. Tfp power 'train-like' directional motion parallel to the long axis of chains of cells, leading to spreading zones around bacteria grown on plates. However, S. sanguinis Tfp are not involved in DNA uptake, which is mediated by a related but distinct nano-machine, and are unusual because they are composed of two pilins in comparable amounts, rather than one as normally seen. Whole genome sequencing identified a locus encoding all the genes involved in Tfp biology in S. sanguinis. A systematic mutational analysis revealed that Tfp biogenesis in S. sanguinis relies on a more basic machinery (only 10 components) than in Gram-negative species and that a small subset of four proteins dispensable for pilus biogenesis are essential for motility. Intriguingly, one of the piliated mutants that does not exhibit spreading retains microscopic motility but moves sideways, which suggests that the corresponding protein controls motion directionality. Besides establishing S. sanguinis as a useful new model for studying Tfp biology, these findings have important implications for our understanding of these widespread filamentous nano-machines. © 2015 The Authors. Molecular Microbiology published by John Wiley & Sons Ltd.

  19. Refining the plasmid-encoded type IV secretion system substrate repertoire of Coxiella burnetii.

    Science.gov (United States)

    Maturana, Pauline; Graham, Joseph G; Sharma, Uma M; Voth, Daniel E

    2013-07-01

    The intracellular bacterial agent of Q fever, Coxiella burnetii, translocates effector proteins into its host cell cytosol via a Dot/Icm type IV secretion system (T4SS). The T4SS is essential for parasitophorous vacuole formation, intracellular replication, and inhibition of host cell death, but the effectors mediating these events remain largely undefined. Six Dot/Icm substrate-encoding genes were recently discovered on the C. burnetii cryptic QpH1 plasmid, three of which are conserved among all C. burnetii isolates, suggesting that they are critical for conserved pathogen functions. However, the remaining hypothetical proteins encoded by plasmid genes have not been assessed for their potential as T4SS substrates. In the current study, we further defined the T4SS effector repertoire encoded by the C. burnetii QpH1, QpRS, and QpDG plasmids that were originally isolated from acute-disease, chronic-disease, and severely attenuated isolates, respectively. Hypothetical proteins, including those specific to QpRS or QpDG, were screened for translocation using the well-established Legionella pneumophila T4SS secretion model. In total, six novel plasmid-encoded proteins were translocated into macrophage-like cells by the Dot/Icm T4SS. Four newly identified effectors are encoded by genes present only on the QpDG plasmid from severely attenuated Dugway isolates, suggesting that the presence of specific effectors correlates with decreased virulence. These results further support the idea of a critical role for extrachromosomal elements in C. burnetii pathogenesis.

  20. Brucella Modulates Secretory Trafficking via Multiple Type IV Secretion Effector Proteins

    Science.gov (United States)

    Myeni, Sebenzile; Child, Robert; Ng, Tony W.; Kupko, John J.; Wehrly, Tara D.; Porcella, Stephen F.; Knodler, Leigh A.; Celli, Jean

    2013-01-01

    The intracellular pathogenic bacterium Brucella generates a replicative vacuole (rBCV) derived from the endoplasmic reticulum via subversion of the host cell secretory pathway. rBCV biogenesis requires the expression of the Type IV secretion system (T4SS) VirB, which is thought to translocate effector proteins that modulate membrane trafficking along the endocytic and secretory pathways. To date, only a few T4SS substrates have been identified, whose molecular functions remain unknown. Here, we used an in silico screen to identify putative T4SS effector candidate proteins using criteria such as limited homology in other bacterial genera, the presence of features similar to known VirB T4SS effectors, GC content and presence of eukaryotic-like motifs. Using β-lactamase and CyaA adenylate cyclase reporter assays, we identified eleven proteins translocated into host cells by Brucella, five in a VirB T4SS-dependent manner, namely BAB1_0678 (BspA), BAB1_0712 (BspB), BAB1_0847 (BspC), BAB1_1671 (BspE) and BAB1_1948 (BspF). A subset of the translocated proteins targeted secretory pathway compartments when ectopically expressed in HeLa cells, and the VirB effectors BspA, BspB and BspF inhibited protein secretion. Brucella infection also impaired host protein secretion in a process requiring BspA, BspB and BspF. Single or combined deletions of bspA, bspB and bspF affected Brucella ability to replicate in macrophages and persist in the liver of infected mice. Taken together, these findings demonstrate that Brucella modulates secretory trafficking via multiple T4SS effector proteins that likely act coordinately to promote Brucella pathogenesis. PMID:23950720

  1. Cooperative retraction of bundled type IV pili enables nanonewton force generation.

    Directory of Open Access Journals (Sweden)

    Nicolas Biais

    2008-04-01

    Full Text Available The causative agent of gonorrhea, Neisseria gonorrhoeae, bears retractable filamentous appendages called type IV pili (Tfp. Tfp are used by many pathogenic and nonpathogenic bacteria to carry out a number of vital functions, including DNA uptake, twitching motility (crawling over surfaces, and attachment to host cells. In N. gonorrhoeae, Tfp binding to epithelial cells and the mechanical forces associated with this binding stimulate signaling cascades and gene expression that enhance infection. Retraction of a single Tfp filament generates forces of 50-100 piconewtons, but nothing is known, thus far, on the retraction force ability of multiple Tfp filaments, even though each bacterium expresses multiple Tfp and multiple bacteria interact during infection. We designed a micropillar assay system to measure Tfp retraction forces. This system consists of an array of force sensors made of elastic pillars that allow quantification of retraction forces from adherent N. gonorrhoeae bacteria. Electron microscopy and fluorescence microscopy were used in combination with this novel assay to assess the structures of Tfp. We show that Tfp can form bundles, which contain up to 8-10 Tfp filaments, that act as coordinated retractable units with forces up to 10 times greater than single filament retraction forces. Furthermore, single filament retraction forces are transient, whereas bundled filaments produce retraction forces that can be sustained. Alterations of noncovalent protein-protein interactions between Tfp can inhibit both bundle formation and high-amplitude retraction forces. Retraction forces build over time through the recruitment and bundling of multiple Tfp that pull cooperatively to generate forces in the nanonewton range. We propose that Tfp retraction can be synchronized through bundling, that Tfp bundle retraction can generate forces in the nanonewton range in vivo, and that such high forces could affect infection.

  2. Peptidomimetic Small Molecules Disrupt Type IV Secretion System Activity in Diverse Bacterial Pathogens

    Directory of Open Access Journals (Sweden)

    Carrie L. Shaffer

    2016-04-01

    Full Text Available Bacteria utilize complex type IV secretion systems (T4SSs to translocate diverse effector proteins or DNA into target cells. Despite the importance of T4SSs in bacterial pathogenesis, the mechanism by which these translocation machineries deliver cargo across the bacterial envelope remains poorly understood, and very few studies have investigated the use of synthetic molecules to disrupt T4SS-mediated transport. Here, we describe two synthetic small molecules (C10 and KSK85 that disrupt T4SS-dependent processes in multiple bacterial pathogens. Helicobacter pylori exploits a pilus appendage associated with the cag T4SS to inject an oncogenic effector protein (CagA and peptidoglycan into gastric epithelial cells. In H. pylori, KSK85 impedes biogenesis of the pilus appendage associated with the cag T4SS, while C10 disrupts cag T4SS activity without perturbing pilus assembly. In addition to the effects in H. pylori, we demonstrate that these compounds disrupt interbacterial DNA transfer by conjugative T4SSs in Escherichia coli and impede vir T4SS-mediated DNA delivery by Agrobacterium tumefaciens in a plant model of infection. Of note, C10 effectively disarmed dissemination of a derepressed IncF plasmid into a recipient bacterial population, thus demonstrating the potential of these compounds in mitigating the spread of antibiotic resistance determinants driven by conjugation. To our knowledge, this study is the first report of synthetic small molecules that impair delivery of both effector protein and DNA cargos by diverse T4SSs.

  3. Carboxyl terminus heterogeneity of type IV fimbrial subunit protein of Pasteurella multocida isolates.

    Science.gov (United States)

    Shivachandra, Sathish Bhadravati; Kumar, Abhinendra; Yogisharadhya, Revanaiah; Ramakrishnan, M A; Viswas, K N

    2013-12-01

    Pasteurella multocida, a Gram-negative bacterial pathogen, known to affect a wide range of domestic as well as wild animal and avian species throughout the world by causing either systemic or localized infections termed as 'pasteurellosis'. P. multocida isolates are known to possess type IV fimbriae (pili) as one of the major virulence factors based on their role in adhesion to host surfaces and subsequent pathogenesis. In the present study, ptfA gene of Indian P. multocida isolates (n = 8) originated from different animal (buffalo, sheep, goat, pig) and avian host species (chicken, turkey, duck, quail) were amplified, cloned, sequenced and compared with available ptfA/fimbrial protein sequences in GenBank/publications (n = 22) to understand its variability with respect to geography/host/serogroup/disease specific patterns. Multiple sequence alignment revealed highly conserved N-terminus α-1 helix region and heterogeneous C-terminus (68-137 aa) comprised of β-strand regions (β1, β2, β3, β4) with conserved two pairs of cysteine residues. Interestingly, an existence of absolute homogeneity among the P. multocida isolates that caused haemorrhagic septicaemia in bovines and septicaemic pasteurellosis in sheep and goats was noticed. Pig isolates had 99.3% homogeneity. On contrary, more diversity (35.8%) was observed among isolates that caused fowl cholera in avians irrespective of identical capsular/somatic serogroup and similar host species. Phylogenetic analysis based on nucleotide sequences of ptfA gene revealed formation of mixed clusters with isolates representing different disease conditions as well as serogroups irrespective of country of origin which indicated the possible role of cross-species transmission among different animal/avian species. The study indicated highly conserved and host specific fimbriae among animal species than relatively divergent fimbriae among avian species.

  4. Properties of type IV plaster considering variation in the mold/model position during setting stage

    Directory of Open Access Journals (Sweden)

    Tarcisio José de Arruda Paes Junior

    2010-04-01

    Full Text Available Objective: To assess the influence of the position of the mold during the setting stage of type IV stone plaster Durone (Dentsply Ind. Com., Rio de Janeiro, Brazil, on the following properties: surface hardness and roughness. Methods: For the roughness test, two groups (n=6 in the form of pellets were prepared. In the first group, the surface of the base of the device was turned down during the plaster setting stage (N, in the second group this position was inverted, which has been described as an act of capsize it (E. For analysis, a roughness meter with reading precision of 0.01 μm was used. With regard to the hardness analysis, two groups with conical-shaped samples were obtained. The plasters were left to set under the same conditions of the mold/model position described for the previous experiment. Hardness measurement was performed in a durometer with a spherical penetrating tip for Rockwell readout. Three measurements were performed for each test specimen in both tests. Results: The hardness (N - 39.8, standard deviation = 3.3, E - 30.8, standard deviation = 5.6 and roughness data (N - 0.67, standard deviation = 0.17, E - 0.74, standard deviation = 0.13 submitted to the Student’s-t test (5% showed no statistically significant differences for the roughness test (0.489, but showed statistically significant differences for the hardness test (0.014. Conclusion: The variation in the mold/model position influenced the final characteristics of the specimens in terms of hardness, since those obtained with the capsize technique showed lower surface hardness, whereas for roughness these differences were not statistically significant.

  5. The Trw type IV secretion system of Bartonella mediates host-specific adhesion to erythrocytes.

    Directory of Open Access Journals (Sweden)

    Muriel Vayssier-Taussat

    2010-06-01

    Full Text Available Bacterial pathogens typically infect only a limited range of hosts; however, the genetic mechanisms governing host-specificity are poorly understood. The alpha-proteobacterial genus Bartonella comprises 21 species that cause host-specific intraerythrocytic bacteremia as hallmark of infection in their respective mammalian reservoirs, including the human-specific pathogens Bartonella quintana and Bartonella bacilliformis that cause trench fever and Oroya fever, respectively. Here, we have identified bacterial factors that mediate host-specific erythrocyte colonization in the mammalian reservoirs. Using mouse-specific Bartonella birtlesii, human-specific Bartonella quintana, cat-specific Bartonella henselae and rat-specific Bartonella tribocorum, we established in vitro adhesion and invasion assays with isolated erythrocytes that fully reproduce the host-specificity of erythrocyte infection as observed in vivo. By signature-tagged mutagenesis of B. birtlesii and mutant selection in a mouse infection model we identified mutants impaired in establishing intraerythrocytic bacteremia. Among 45 abacteremic mutants, five failed to adhere to and invade mouse erythrocytes in vitro. The corresponding genes encode components of the type IV secretion system (T4SS Trw, demonstrating that this virulence factor laterally acquired by the Bartonella lineage is directly involved in adherence to erythrocytes. Strikingly, ectopic expression of Trw of rat-specific B. tribocorum in cat-specific B. henselae or human-specific B. quintana expanded their host range for erythrocyte infection to rat, demonstrating that Trw mediates host-specific erythrocyte infection. A molecular evolutionary analysis of the trw locus further indicated that the variable, surface-located TrwL and TrwJ might represent the T4SS components that determine host-specificity of erythrocyte parasitism. In conclusion, we show that the laterally acquired Trw T4SS diversified in the Bartonella lineage

  6. Colonoscopic perforation leading to a diagnosis of Ehlers Danlos syndrome type IV: a case report and review of the literature

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    Wolfe John

    2011-06-01

    Full Text Available Abstract Introduction Colonoscopic perforation is a rare but serious complication of colonoscopy. Factors known to increase the risk of perforation include colonic strictures, extensive diverticulosis, and friable tissues. We describe the case of a man who was found to have perforation of the sigmoid colon secondary to an undiagnosed connective tissue disorder (Ehlers-Danlos syndrome type IV while undergoing surveillance for hereditary non-polyposis colorectal cancer. Case presentation A 33-year-old Caucasian man presented to our hospital with an acute abdomen following a colonoscopy five days earlier as part of hereditary non-polyposis colorectal cancer screening. His medical history included bilateral clubfoot. His physical examination findings suggested left iliac fossa peritonitis. A computed tomographic scan revealed perforation of the sigmoid colon and incidentally a right common iliac artery aneurysm as well. Hartmann's procedure was performed during laparotomy. The patient recovered well post-operatively and was discharged. Reversal of the Hartmann's procedure was performed six months later. This procedure was challenging because of dense adhesions and friable bowel. The histology of bowel specimens from this surgery revealed thinning and fibrosis of the muscularis externa. The patient was subsequently noted to have transparency of truncal skin with easily visible vessels. An underlying collagen vascular disorder was suspected, and genetic testing revealed a mutation in the collagen type III, α1 (COL3A1 gene, which is consistent with a diagnosis of Ehlers-Danlos syndrome type IV. Conclusions Ehlers-Danlos syndrome type IV, the vascular type, is a rare disorder caused by mutations in the COL3A1 gene on chromosome 2q31. It is characterized by translucent skin, clubfoot, and the potentially fatal complications of spontaneous large vessel rupture, although spontaneous uterine and colonic perforations have also been reported in the

  7. Crescentic glomerulonephritis and subepidermal blisters with autoantibodies to alpha5 and alpha6 chains of type IV collagen.

    Science.gov (United States)

    Ghohestani, Reza F; Rotunda, Sherry L; Hudson, Billy; Gaughan, William J; Farber, John L; Webster, Guy; Uitto, Jouni

    2003-05-01

    We describe a novel autoimmune disease characterized by severe subepidermal bullous eruption and crescentic glomerulonephritis with autoantibodies directed against the noncollagenous domain of the alpha5 and alpha6 chains of type IV collagen. Biopsy of perilesional skin revealed a subepidermal blister with marked polymorphonuclear infiltrate with linear deposits of IgA and C3. Light microscopy of a kidney biopsy specimen revealed a crescentic glomerulonephritis, and immunofluorescence microscopy showed linear basement membrane staining for IgA (3+), C3 (1+), and IgG (1+). No electron-dense deposits were observed by transmission electron microscopy. The patient's autoantibodies reacted with normal human skin and kidney: IgA (3+) and IgG (1+) antibodies stained the basement membrane zones of skin, renal glomerulus, and some tubules. The identity of the target antigen was determined by immunochemical analyses of candidate antigens using the patient's autoantibodies. The patient's IgA and IgG autoantibodies reacted with a 185- to 190-kDa antigen from a human dermal extract that was distinguished from the other dermal or epidermal antigens, including the 145- to 290-kDa (type VII collagen) epidermolysis bullosa acquisita antigen, the 165- to 200-kDa alpha3 laminin mucous membrane cicatricial pemphigoid antigen, and the 230-kDa and the 180-kDa bullous pemphigoid antigens. Patient's IgA and IgG autoantibodies further reacted with the alpha5(IV) and weakly with the alpha6(IV) chains of type IV collagen by Western blot and ELISA. This report expands the repertoire of bullous skin disorders and provides an explanation for the association of anti-type IV collagen autoantibodies and glomerulonephritis with subepidermal blisters.

  8. Spine deformities in patients with Ehlers-Danlos syndrome, type IV - late results of surgical treatment

    Directory of Open Access Journals (Sweden)

    Tesiorowski Maciej

    2010-11-01

    Full Text Available Abstract Background Spinal deformities in Ehlers-Danlos syndrome are usually progressive and may require operative treatment. There is limited number of studies describing late results of surgery in this disease. Methods This is a retrospective study of the records of 11 patients with Ehlers-Danlos syndrome type IV, treated surgically between 1990 and 2007. All patients underwent surgical treatment for spinal deformity. Duration of operation, type of instrumentation, intraoperative blood loss, complications and number of additional surgeries were noted. Radiographic measurement was performed on standing AP and lateral radiographs acquired before surgery, just after and at final follow up. Results The mean follow up period was 5.5 ± 2.9 years (range 1-10 years. The mean preoperative thoracic and lumbar curve were 109.5 ± 19.9° (range 83° - 142° and 75.6 ± 26.7° (range 40° - 108° respectively. Posterior spine fusion alone was performed on 6 patients and combined anterior and posterior fusion (one- or two stage on 5 cases. Posterior segmental spinal instrumentation was applied with use of hooks, screws and wires. The mean postoperative thoracic and lumbar curve improved to 79.3 ± 16.1° (range 56° - 105° and 58.5 ± 27.7° (range 10° - 95° respectively, with a slight loss of correction during follow up. The average thoracic and lumbar correction was 26.4 ± 14.9% (range 5.3 - 50.4% and 26.3 ± 21.2% (range 7.9 - 75%. Postoperatively, the mean kyphosis was 79.5 ± 40.3° (range 21° -170°, and lordosis was 50.8 ± 18.6° (range 20° -79°. Hyperkyphosis increased during follow up while lordosis remained stable. Mean Th12-L2 angle was -3.5 ±9.9° (range -19° - 15° postoperatively and did not change significantly during follow up. Conclusions Huge spinal deformities in patients with Ehlers-Danlos syndrome require complex and extensive surgery. There is a big risk of sagittal imbalance in this group.

  9. Essential components of the Ti plasmid trb system, a type IV macromolecular transporter.

    Science.gov (United States)

    Li, P L; Hwang, I; Miyagi, H; True, H; Farrand, S K

    1999-08-01

    The trb operon from pTiC58 is one of three loci that are required for conjugal transfer of this Ti plasmid. The operon, which probably codes for the mating bridge responsible for pair formation and DNA transfer, contains 12 genes, 11 of which are related to genes from other members of the type IV secretion system family. The 12th gene, traI, codes for production of Agrobacterium autoinducer (AAI). Insertion mutations were constructed in each of the 12 genes, contained on a full-length clone of the trb region, using antibiotic resistance cassettes or a newly constructed transposon. This transposon, called mini-Tn5Ptrb, was designed to express genes downstream of the insertion site from a promoter regulated by TraR and AAI. Each mutation could trans complement downstream Tn3HoHo1 insertions in the trb operon of full-sized Ti plasmids. When marker-exchanged into the transfer-constitutive Ti plasmid pTiC58DeltaaccR mutations in trbB, -C, -D, -E, -L, -F, -G, and -H abolished conjugal transfer from strain UIA5, which lacks the 450-kb catabolic plasmid pAtC58. However, these mutants retained residual conjugal transfer activity when tested in strain NT1, which contains this large plasmid. The trbJ mutant failed to transfer at a detectable frequency from either strain, while the trbI mutant transferred at very low but detectable levels from both donors. Only the trbK mutant was unaffected in conjugal transfer from either donor. Transfer of each of the marker-exchange mutants was restored by a clone expressing only the wild-type allele of the corresponding mutant trb gene. An insertion mutation in traI abolished the production of AAI and also conjugal transfer. This defect was restored by culturing the mutant donor in the presence of AAI. We conclude that all of the trb genes except trbI and trbK are essential for conjugal transfer of pTiC58. We also conclude that mutations in any one of the trb genes except traI and trbJ can be complemented by functions coded for by pAtC58.

  10. Spine deformities in patients with Ehlers-Danlos syndrome, type IV - late results of surgical treatment

    Science.gov (United States)

    2010-01-01

    Background Spinal deformities in Ehlers-Danlos syndrome are usually progressive and may require operative treatment. There is limited number of studies describing late results of surgery in this disease. Methods This is a retrospective study of the records of 11 patients with Ehlers-Danlos syndrome type IV, treated surgically between 1990 and 2007. All patients underwent surgical treatment for spinal deformity. Duration of operation, type of instrumentation, intraoperative blood loss, complications and number of additional surgeries were noted. Radiographic measurement was performed on standing AP and lateral radiographs acquired before surgery, just after and at final follow up. Results The mean follow up period was 5.5 ± 2.9 years (range 1-10 years). The mean preoperative thoracic and lumbar curve were 109.5 ± 19.9° (range 83° - 142°) and 75.6 ± 26.7° (range 40° - 108°) respectively. Posterior spine fusion alone was performed on 6 patients and combined anterior and posterior fusion (one- or two stage) on 5 cases. Posterior segmental spinal instrumentation was applied with use of hooks, screws and wires. The mean postoperative thoracic and lumbar curve improved to 79.3 ± 16.1° (range 56° - 105°) and 58.5 ± 27.7° (range 10° - 95°) respectively, with a slight loss of correction during follow up. The average thoracic and lumbar correction was 26.4 ± 14.9% (range 5.3 - 50.4%) and 26.3 ± 21.2% (range 7.9 - 75%). Postoperatively, the mean kyphosis was 79.5 ± 40.3° (range 21° -170°), and lordosis was 50.8 ± 18.6° (range 20° -79°). Hyperkyphosis increased during follow up while lordosis remained stable. Mean Th12-L2 angle was -3.5 ±9.9° (range -19° - 15°) postoperatively and did not change significantly during follow up. Conclusions Huge spinal deformities in patients with Ehlers-Danlos syndrome require complex and extensive surgery. There is a big risk of sagittal imbalance in this group. PMID:21108838

  11. Quantitative analysis of immunogold labellings of collagen types I, III, IV and VI in healthy and pathological human corneas.

    Science.gov (United States)

    Delaigue, O; Arbeille, B; Rossazza, C; Lemesle, M; Roingeard, P

    1995-06-01

    We studied the distribution of collagen types I, III, IV and VI in one healthy human cornea and in seven pathological human corneas, in which the disorders were three cases of pseudophakic bullous keratopathy (two severe, one moderate) and one case each of stage IV keratoconus, chronic ulcer, vascularized cornea and disciform keratitis. Transmission electron microscopy examinations were performed on post-embedding immunogold-labelled sections. The staining was evaluated by gold particle count in the different tissues. The presence or absence of a given antigen was determined by statistical analysis, using a d-value test. Our results on healthy corneal tissues corroborate the data available from previous studies, except for collagen type VI, which we found to be absent in Bowman's layer. In pathological corneas with a collagenous layer posterior to Descemet's membrane, collagen types I, III and especially IV were detected in this collagenous layer. Collagen types I, III and VI were detected in the anterior healed stroma of other pathological corneas, except for the keratoconus cornea, in which intense collagen III staining was observed. The presence of collagen types I and III in the posterior collagenous layer of our pseudophakic bullous keratopathy corneas suggests that this layer corresponds to scar tissue secreted by stimulated endothelial cells.

  12. Novel frameshift and splice site mutations in the neurotrophic tyrosine kinase receptor type 1 gene (NTRK1) associated with hereditary sensory neuropathy type IV.

    Science.gov (United States)

    Verpoorten, Nathalie; Claeys, Kristl G; Deprez, Liesbet; Jacobs, An; Van Gerwen, Veerle; Lagae, Lieven; Arts, Willem Frans; De Meirleir, Linda; Keymolen, Kathelijn; Ceuterick-de Groote, Chantal; De Jonghe, Peter; Timmerman, Vincent; Nelis, Eva

    2006-01-01

    Congenital insensitivity to pain with anhidrosis or hereditary sensory and autonomic neuropathy type IV (HSAN IV) is the first human genetic disorder implicated in the neurotrophin signal transduction pathway. HSAN IV is characterized by absence of reaction to noxious stimuli, recurrent episodes of fever, anhidrosis, self-mutilating behavior and often mental retardation. Mutations in the neurotrophic tyrosine kinase, receptor, type 1 (NTRK1) are associated with this disorder. Here we report four homozygous mutations, two frameshift (p.Gln626fsX6 and p.Gly181fsX58), one missense (p.Arg761Trp) and one splice site (c.359+5G>T) mutation in four HSAN IV patients. The splice site mutation caused skipping of exons 2 and 3 in patient's mRNA resulting in an in-frame deletion of the second leucine-rich motif. NTRK1 mutations are only rarely reported in the European population. This report extends the spectrum of NTRK1 mutations observed in patients diagnosed with HSAN IV.

  13. Comparative Genomics of the Zoonotic Pathogen Ehrlichia chaffeensis Reveals Candidate Type IV Effectors and Putative Host Cell Targets.

    Science.gov (United States)

    Noroy, Christophe; Meyer, Damien F

    2016-01-01

    During infection, some intracellular pathogenic bacteria use a dedicated multiprotein complex known as the type IV secretion system to deliver type IV effector (T4E) proteins inside the host cell. These T4Es allow the bacteria to evade host defenses and to subvert host cell processes to their own advantage. Ehrlichia chaffeensis is a tick-transmitted obligate intracellular pathogenic bacterium, which causes human monocytic ehrlichiosis. Using comparative whole genome analysis, we identified the relationship between eight available E. chaffeensis genomes isolated from humans and show that these genomes are highly conserved. We identified the candidate core type IV effectome of E. chaffeensis and some conserved intracellular adaptive strategies. We assigned the West Paces strain to genetic group II and predicted the repertoires of T4Es encoded by E. chaffeensis genomes, as well as some putative host cell targets. We demonstrated that predicted T4Es are preferentially distributed in gene sparse regions of the genome. In addition to the identification of the two known type IV effectors of Anaplasmataceae, we identified two novel candidates T4Es, ECHLIB_RS02720 and ECHLIB_RS04640, which are not present in all E. chaffeensis strains and could explain some variations in inter-strain virulence. We also identified another novel candidate T4E, ECHLIB_RS02720, a hypothetical protein exhibiting EPIYA, and NLS domains as well as a classical type IV secretion signal, suggesting an important role inside the host cell. Overall, our results agree with current knowledge of Ehrlichia molecular pathogenesis, and reveal novel candidate T4Es that require experimental validation. This work demonstrates that comparative effectomics enables identification of important host pathways targeted by the bacterial pathogen. Our study, which focuses on the type IV effector repertoires among several strains of E. chaffeensis species, is an original approach and provides rational putative targets

  14. Implementation of GLP-1 based therapy of type 2 diabetes mellitus using DPP-IV inhibitors

    DEFF Research Database (Denmark)

    Holst, Jens Juul

    2003-01-01

    GLP-1 is a peptide hormone from the intestinal mucosa. It is secreted in response to meal ingestion and normally functions in the so-called ileal brake i. e. inhibition of upper gastrointestinal motility and secretion when nutrients are present in the distal small intestine. It also induces satie......, it is as yet uncertain wether DDP-IV inhibitors will affect gastrointestinal motility, appetite and food intake. Even the effects of GLP-1 effects on the pancreatic islets may be partly neurally mediated and therefore uninfluenced by DPP-IV inhibition....

  15. An anomalous type IV secretion system in Rickettsia is evolutionarily conserved.

    Directory of Open Access Journals (Sweden)

    Joseph J Gillespie

    Full Text Available BACKGROUND: Bacterial type IV secretion systems (T4SSs comprise a diverse transporter family functioning in conjugation, competence, and effector molecule (DNA and/or protein translocation. Thirteen genome sequences from Rickettsia, obligate intracellular symbionts/pathogens of a wide range of eukaryotes, have revealed a reduced T4SS relative to the Agrobacterium tumefaciens archetype (vir. However, the Rickettsia T4SS has not been functionally characterized for its role in symbiosis/virulence, and none of its substrates are known. RESULTS: Superimposition of T4SS structural/functional information over previously identified Rickettsia components implicate a functional Rickettsia T4SS. virB4, virB8 and virB9 are duplicated, yet only one copy of each has the conserved features of similar genes in other T4SSs. An extraordinarily duplicated VirB6 gene encodes five hydrophobic proteins conserved only in a short region known to be involved in DNA transfer in A. tumefaciens. virB1, virB2 and virB7 are newly identified, revealing a Rickettsia T4SS lacking only virB5 relative to the vir archetype. Phylogeny estimation suggests vertical inheritance of all components, despite gene rearrangements into an archipelago of five islets. Similarities of Rickettsia VirB7/VirB9 to ComB7/ComB9 proteins of epsilon-proteobacteria, as well as phylogenetic affinities to the Legionella lvh T4SS, imply the Rickettsiales ancestor acquired a vir-like locus from distantly related bacteria, perhaps while residing in a protozoan host. Modern modifications of these systems likely reflect diversification with various eukaryotic host cells. CONCLUSION: We present the rvh (Rickettsiales vir homolog T4SS, an evolutionary conserved transporter with an unknown role in rickettsial biology. This work lays the foundation for future laboratory characterization of this system, and also identifies the Legionella lvh T4SS as a suitable genetic model.

  16. Antimicrobial peptide inhibition of fungalysin proteases that target plant type 19 Family IV defense chitinases

    Science.gov (United States)

    Cereal crops and other plants produce secreted seed chitinases that reduce pathogenic infection, most likely by targeting the fungal chitinous cell wall. We have shown that corn (Zea mays) produces three GH family 19, plant class IV chitinases, that help in protecting the plant against Fusarium and ...

  17. Familial Amyloid Polyneuropathy Type IV (FINNISH with Rapid Clinical Progression in an Iranian Woman: A Case Report

    Directory of Open Access Journals (Sweden)

    Arash Babaei-Ghazani

    2016-05-01

    Full Text Available Familial amyloid polyneuropathy (FAP type IV (FINNISH is a rare clinical entity with challenging neuropathy and cosmetic deficits. Amyloidosis can affect peripheral sensory, motor, or autonomic nerves. Nerve lesions are induced by deposits of amyloid fibrils and treatment approaches for neuropathy are challenging. Involvement of cranial nerves and atrophy in facial muscles is a real concern in daily life of such patients. Currently, diagnosis of neuropathy can be made by electrodiagnostic studies and diagnosis of amyloidosis can be made by genetic testing or by detection of amyloid deposition in abdominal fat pad, rectal, or nerve biopsies. It is preferable to consider FAP as one of the differential diagnosis of a case presented with multiple cranial nerves symptoms. The authors present a case of familial amyloid polyneuropathy (FAP type IV with severe involvement of multiple cranial nerves, peripheral limb neuropathy, and orthostatic hypotension.

  18. Steered molecular dynamics simulations of a type IV pilus probe initial stages of a force-induced conformational transition.

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    Joseph L Baker

    2013-04-01

    Full Text Available Type IV pili are long, protein filaments built from a repeating subunit that protrudes from the surface of a wide variety of infectious bacteria. They are implicated in a vast array of functions, ranging from bacterial motility to microcolony formation to infection. One of the most well-studied type IV filaments is the gonococcal type IV pilus (GC-T4P from Neisseria gonorrhoeae, the causative agent of gonorrhea. Cryo-electron microscopy has been used to construct a model of this filament, offering insights into the structure of type IV pili. In addition, experiments have demonstrated that GC-T4P can withstand very large tension forces, and transition to a force-induced conformation. However, the details of force-generation, and the atomic-level characteristics of the force-induced conformation, are unknown. Here, steered molecular dynamics (SMD simulation was used to exert a force in silico on an 18 subunit segment of GC-T4P to address questions regarding the nature of the interactions that lead to the extraordinary strength of bacterial pili. SMD simulations revealed that the buried pilin α1 domains maintain hydrophobic contacts with one another within the core of the filament, leading to GC-T4P's structural stability. At the filament surface, gaps between pilin globular head domains in both the native and pulled states provide water accessible routes between the external environment and the interior of the filament, allowing water to access the pilin α1 domains as reported for VC-T4P in deuterium exchange experiments. Results were also compared to the experimentally observed force-induced conformation. In particular, an exposed amino acid sequence in the experimentally stretched filament was also found to become exposed during the SMD simulations, suggesting that initial stages of the force induced transition are well captured. Furthermore, a second sequence was shown to be initially hidden in the native filament and became exposed upon

  19. Two-stage hybrid open-endovascular repair of a Crawford type IV aortic aneurysm: therapeutic challenge

    Directory of Open Access Journals (Sweden)

    Abdo Farret Neto

    2014-04-01

    Full Text Available We present a case of a patient with Crawford type IV aortic thoracoabdominal aneurysm. The patient underwent hybrid repair in two stages. Initially a Dacron(r graft was implanted surgically with revascularization of all visceral branches from the left external iliac artery (debranching. On a later date, the second stage of treatment was with an endovascular technique with bi aorto-iliac endoprosthesis. After 2 years the patient remains asymptomatic and in full working activity.

  20. Polyvinylidene fluoride for proliferation and preservation of bovine corneal endothelial cells by enhancing type IV collagen production and deposition.

    Science.gov (United States)

    Wang, Tsung-Jen; Wang, I-Jong; Chen, Yi-Hsin; Lu, Jui-Nan; Young, Tai-Horng

    2012-01-01

    In this study, biomaterials with different hydrophobic properties including polyvinyl alcohol (PVA), poly(ethylene-co-vinyl alcohol) (EVAL), tissue culture polystyrene (TCPS), and polyvinylidene fluoride (PVDF) were examined in the bovine corneal endothelial cells (BCECs) culture system to elucidate their possible impact on clinical demand and scientific interest. It was found that BCECs were inhibited to attach onto the PVA surface. Conversely, relatively more hydrophobic biomaterials EVAL, TCPS, and PVDF successfully initiate BCEC adhesion. Compared to EVAL, cultured BCECs on TCPS and PVDF exhibited higher viability. Furthermore, fibroblastic transformation on EVAL and TCPS was observed at day 17, but BCECs maintained typical hexagonal shape on the PVDF surface at day 21. This phenomenon can be rescued by previously coating type IV collagen on TCPS but not on EVAL. In addition, when BCECs were cultured on PVDF, the expressions of gap junction connexin-43, differentiation marker N-cadherin, and tight junction ZO-1 were well-developed, resembling the physiological phenotypes. After examining the type IV collagen expression by Western blot analysis and protein absorption test, a possible explanation for the better proliferation and preservation of BCECs on the PVDF substrate is that PVDF is a bioactive substratum which enables BCECs to synthesize and reserve more extracellular matrix type IV collagen, paving an important way to provide a more preferential environment for BCEC cultures. Accordingly, promoting CEC growth effects after cell-biomaterial association may be applied to the tissue engineering of corneal endothelium. Copyright © 2011 Wiley Periodicals, Inc.

  1. DNA Delivery and Genomic Integration into Mammalian Target Cells through Type IV A and B Secretion Systems of Human Pathogens.

    Science.gov (United States)

    Guzmán-Herrador, Dolores L; Steiner, Samuel; Alperi, Anabel; González-Prieto, Coral; Roy, Craig R; Llosa, Matxalen

    2017-01-01

    We explore the potential of bacterial secretion systems as tools for genomic modification of human cells. We previously showed that foreign DNA can be introduced into human cells through the Type IV A secretion system of the human pathogen Bartonella henselae. Moreover, the DNA is delivered covalently attached to the conjugative relaxase TrwC, which promotes its integration into the recipient genome. In this work, we report that this tool can be adapted to other target cells by using different relaxases and secretion systems. The promiscuous relaxase MobA from plasmid RSF1010 can be used to deliver DNA into human cells with higher efficiency than TrwC. MobA also promotes DNA integration, albeit at lower rates than TrwC. Notably, we report that DNA transfer to human cells can also take place through the Type IV secretion system of two intracellular human pathogens, Legionella pneumophila and Coxiella burnetii, which code for a distantly related Dot/Icm Type IV B secretion system. This suggests that DNA transfer could be an intrinsic ability of this family of secretion systems, expanding the range of target human cells. Further analysis of the DNA transfer process showed that recruitment of MobA by Dot/Icm was dependent on the IcmSW chaperone, which may explain the higher DNA transfer rates obtained. Finally, we observed that the presence of MobA negatively affected the intracellular replication of C. burnetii, suggesting an interference with Dot/Icm translocation of virulence factors.

  2. Type IV secretion-dependent activation of host MAP kinases induces an increased proinflammatory cytokine response to Legionella pneumophila.

    Directory of Open Access Journals (Sweden)

    Sunny Shin

    2008-11-01

    Full Text Available The immune system must discriminate between pathogenic and nonpathogenic microbes in order to initiate an appropriate response. Toll-like receptors (TLRs detect microbial components common to both pathogenic and nonpathogenic bacteria, whereas Nod-like receptors (NLRs sense microbial components introduced into the host cytosol by the specialized secretion systems or pore-forming toxins of bacterial pathogens. The host signaling pathways that respond to bacterial secretion systems remain poorly understood. Infection with the pathogen Legionella pneumophila, which utilizes a type IV secretion system (T4SS, induced an increased proinflammatory cytokine response compared to avirulent bacteria in which the T4SS was inactivated. This enhanced response involved NF-kappaB activation by TLR signaling as well as Nod1 and Nod2 detection of type IV secretion. Furthermore, a TLR- and RIP2-independent pathway leading to p38 and SAPK/JNK MAPK activation was found to play an equally important role in the host response to virulent L. pneumophila. Activation of this MAPK pathway was T4SS-dependent and coordinated with TLR signaling to mount a robust proinflammatory cytokine response to virulent L. pneumophila. These findings define a previously uncharacterized host response to bacterial type IV secretion that activates MAPK signaling and demonstrate that coincident detection of multiple bacterial components enables immune discrimination between virulent and avirulent bacteria.

  3. Genomic and Gene-Expression Comparisons among Phage-Resistant Type-IV Pilus Mutants of Pseudomonas syringae pathovar phaseolicola.

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    Mark Sistrom

    Full Text Available Pseudomonas syringae pv. phaseolicola (Pph is a significant bacterial pathogen of agricultural crops, and phage Φ6 and other members of the dsRNA virus family Cystoviridae undergo lytic (virulent infection of Pph, using the type IV pilus as the initial site of cellular attachment. Despite the popularity of Pph/phage Φ6 as a model system in evolutionary biology, Pph resistance to phage Φ6 remains poorly characterized. To investigate differences between phage Φ6 resistant Pph strains, we examined genomic and gene expression variation among three bacterial genotypes that differ in the number of type IV pili expressed per cell: ordinary (wild-type, non-piliated, and super-piliated. Genome sequencing of non-piliated and super-piliated Pph identified few mutations that separate these genotypes from wild type Pph--and none present in genes known to be directly involved in type IV pilus expression. Expression analysis revealed that 81.1% of gene ontology (GO terms up-regulated in the non-piliated strain were down-regulated in the super-piliated strain. This differential expression is particularly prevalent in genes associated with respiration--specifically genes in the tricarboxylic acid cycle (TCA cycle, aerobic respiration, and acetyl-CoA metabolism. The expression patterns of the TCA pathway appear to be generally up and down-regulated, in non-piliated and super-piliated Pph respectively. As pilus retraction is mediated by an ATP motor, loss of retraction ability might lead to a lower energy draw on the bacterial cell, leading to a different energy balance than wild type. The lower metabolic rate of the super-piliated strain is potentially a result of its loss of ability to retract.

  4. A novel arctigenin-containing latex glove prevents latex allergy by inhibiting type I/IV allergic reactions.

    Science.gov (United States)

    Wang, Yong-Xin; Xue, Dan-Ting; Liu, Meng; Zhou, Zheng-Min; Shang, Jing

    2016-03-01

    The present study aimed at developing a natural compound with anti-allergic effect and stability under latex glove manufacturing conditions and investigating whether its anti-allergic effect is maintained after its addition into the latex. The effects of nine natural compounds on growth of the RBL-2H3 cells and mouse primary spleen lymphocytes were determined using MTT assay. The compounds included glycyrrhizin, osthole, tetrandrine, tea polyphenol, catechin, arctigenin, oleanolic acid, baicalin and oxymatrine. An ELISA assay was used for the in vitro anti-type I/IV allergy screening; in this process β-hexosaminidase, histamine, and IL-4 released from RBL-2H3 cell lines and IFN-γ and IL-2 released from mouse primary spleen lymphocytes were taken as screening indices. The physical stability of eight natural compounds and the dissolubility of arctigenin, selected based on the in vitro pharnacodynamaic screening and the stability evaluation, were detected by HPLC. The in vivo pharmacodynamic confirmation of arctigenin and final latex product was evaluated with a passive cutaneous anaphylaxis (PCA) model and an allergen-specific skin response model. Nine natural compounds showed minor growth inhibition on RBL-2H3 cells and mouse primary spleen lymphocytes. Baicalin and arctigenin had the best anti-type I and IV allergic effects among the natural compounds based on the in vitro pharmacodynamic screening. Arctigenin and catechin had the best physical stability under different manufacturing conditions. Arctigenin was the selected for further evaluation and proven to have anti-type I and IV allergic effects in vivo in a dose-dependent manner. The final product of the arctigenin-containing latex glove had anti-type I and IV allergic effects in vivo which were mainly attributed to arctigenin as proved from the dissolubility results. Arctigenin showed anti-type I and IV allergic effects in vitro and in vivo, with a good stability under latex glove manufacturing conditions

  5. Mapping Type IV Secretion Signals on the Primase Encoded by the Broad-Host-Range Plasmid R1162 (RSF1010).

    Science.gov (United States)

    Meyer, Richard

    2015-10-01

    The plasmid R1162 (RSF1010) encodes a primase essential for its replication. This primase makes up the C-terminal part of MobA, a multifunctional protein with the relaxase as a separate N-terminal domain. The primase is also translated separately as the protein RepB'. Here, we map two signals for type IV secretion onto the recently solved structure of RepB'. One signal is located internally within RepB' and consists of a long α-helix and an adjacent disordered region rich in arginines. The second signal is made up of the same α-helix and a second, arginine-rich region at the C-terminal end of the protein. Successive arginine-to-alanine substitutions revealed that either signal can be utilized by the type IV secretion complex of the plasmid R751. The internal signal also enables conjugal transfer when linked to the relaxase part of MobA. Both signals are similar to those previously identified for type IV secretion substrates in the Vir system of Agrobacterium tumefaciens. Moreover, the C-terminal arginine-rich segment of RepB' has been shown to be secreted by Vir. However, with R751, the signals require MobB, an R1162-encoded accessory protein active in conjugal transfer. The results of two-hybrid assays revealed that MobB interacts, via its membrane-associated domain, with the R751 plasmid coupling protein TraG. In addition, MobB interacts with a region of MobA just outside the RepB' domain. Therefore, MobB is likely an adaptor that is essential for recognition of the primase-associated signals by the R751 secretion machinery. For most plasmids, type IV secretion is an intrinsic part of the mechanism for conjugal transfer. Protein relaxases, bound to the 5' end of the transferring strand, are mobilized into recipient cells by the type IV pathway. In this work, we identify and characterize two signals for secretion in the primase domain of MobA, the relaxase of the IncQ plasmid R1162 (RSF1010). We also show that the adaptor protein MobB is required for engagement

  6. Endovascular treatment for a spontaneous rupture of the posterior tibial artery in a patient with Ehlers-Danlos syndrome Type IV: report of a case.

    Science.gov (United States)

    Matsushima, Kazuhide; Takara, Hiroaki

    2009-01-01

    This report presents the case of a 27-year-old woman with a history of Ehlers-Danlos syndrome Type IV (EDS-IV) who suffered from acute compartment syndrome caused by the spontaneous rupture of the posterior tibial artery. Bleeding control was achieved by endovascular embolization. A fasciotomy was successfully performed without any trouble with hemorrhaging. Endovascular treatment could therefore represent the ideal option for treating infrapopliteal lesions, particularly for EDS-IV patients.

  7. The use of spa and phage typing for characterization of clinical isolates of methicillin-resistant Staphylococcus aureus in the University Clinical Center in Gdańsk, Poland.

    Science.gov (United States)

    Wiśniewska, Katarzyna; Szewczyk, Anna; Piechowicz, Lidia; Bronk, Marek; Samet, Alfred; Swieć, Krystyna

    2012-05-01

    The emergence of spa types and spa-clonal complexes (CC) among clinical methicillin-resistant Staphylococcus aureus isolates collected from the University Clinical Center in Gdańsk between 2008 and 2009 were investigated. Phage typing was used as the initial screening in the study. The basic set of phages and the additional set of phages were used. Most of the isolates (56 %) belonged to the phage group III. With the additional set of phages, eight types were found, with predominant one MR8 (50 %). Sixteen distinct spa types were observed. The most frequent were t003 (22 %), t151 (16 %), and t008 (12 %). The spa types were clustered into two spa-CC and eight singletons. The predominant CC010 (50 %) consisted of six types, with the most common t003 (36.7 %) and t151(26.7 %), and in 80 % was identified as staphylococcal chromosomal casette mec (SCCmec) type II. The second cluster has no founder (12 %) with only two spa types: t037 belonging to SCCmec type III and t029. In the most frequent singleton, spa type t008 alone was clustered in 12 % of the isolates. All singletons correspond to SCCmec type IV. The CC010 was distributed in most of the hospital wards, corresponded to Multilocus sequence typing type ST5/ST225 and was constantly present throughout the observed period. The isolates of CC010 generally belonged to the phage group III, and most of them (53.3 %) were resistant to erythromycin, clindamycin, and ciprofloxacin. The concordance between spa-clone and phage type was very high, but the same phage type MR8 was observed within different spa types of the predominant clone.

  8. Early venous manifestation of Ehlers-Danlos syndrome Type IV through a novel mutation in COL3A1.

    Science.gov (United States)

    Wendorff, Heiko; Pelisek, Jaroslav; Zimmermann, Alexander; Mayer, Karin; Seidel, Heide; Weirich, Gregor; Hausser, Ingrid; Siegel, Corinna; Zernecke, Alma; Eckstein, Hans-Henning

    2013-01-01

    Ehlers-Danlos syndrome (EDS) leads to abnormalities in the synthesis of collagen and complications involving arterial vessels. We describe here a mutation in the intron 14 of the COL3A1 gene leading to EDS Type IV (EDS IV) associated with venous manifestations only. The patient, an 18-year-old male, suffered from truncal varicosity of the long saphenous vein on both sides. Conventional stripping surgery of the left saphenous vein revealed an extremely vulnerable ectatic superficial femoral vein. An inserted vein graft occluded, and venous thrombectomy was unsuccessful. A conservative anticoagulant and compression therapy finally succeeded. This is the first report describing EDS IV due to a mutation in intron 14 of the COL3A1 gene leading to venous manifestations without affecting arterial vessels at clinical presentation. Our findings imply that molecular genetic analysis should be considered in patients with unusual clinical presentation and that conservative therapy should be applied until a suspected clinical diagnosis has been secured. Copyright © 2013 Elsevier Inc. All rights reserved.

  9. Glycogen Storage Disease Type IV and Early Implantation Defect: Early Trophoblastic Involvement Associated with a New GBE1 Mutation.

    Science.gov (United States)

    Dainese, Linda; Adam, Nicolas; Boudjemaa, Sabah; Hadid, Kamel; Rosenblatt, Jonathan; Jouannic, Jean-Marie; Heron, Delphine; Froissart, Roseline; Coulomb, Aurore

    A 29-year-old primigravida presented with a spontaneous miscarriage at 8 weeks of gestation. There was no consanguinity in the family. Aspiration was performed. Pathological examination showed immature villi with numerous slightly yellow intracytoplasmic inclusions within the early implantation stage cytotrophoblastic cells. Inclusions were periodic acid-Schiff and Alcian blue positive and partially positive with periodic acid-Schiff with amylase. Diagnosis of Glycogen storage disease type IV (GSD IV) was made. Genetic analysis of glycogen branching enzyme 1 gene (GBE1) was performed in parents and showed a novel deletion of 1 nucleotide, c.1937delT, affecting the mother and a mutation affecting a consensus splice site, c.691+2T>C, in the father. At time of subsequent pregnancy, genetic counseling with GBE1 gene analysis was performed on throphoblastic biopsy and showed a mutated allele, c.1937delT, inherited from the mother. The mother gave birth to a healthy, unaffected female newborn. Our findings demonstrate that GSD IV may affect early pregnancies, leading to trophoblastic damage and early fetal loss. Diagnosis can accurately be made on pathological examination and should be further documented by genetic analysis.

  10. T346Hunter: a novel web-based tool for the prediction of type III, type IV and type VI secretion systems in bacterial genomes.

    Directory of Open Access Journals (Sweden)

    Pedro Manuel Martínez-García

    Full Text Available T346Hunter (Type Three, Four and Six secretion system Hunter is a web-based tool for the identification and localisation of type III, type IV and type VI secretion systems (T3SS, T4SS and T6SS, respectively clusters in bacterial genomes. Non-flagellar T3SS (NF-T3SS and T6SS are complex molecular machines that deliver effector proteins from bacterial cells into the environment or into other eukaryotic or prokaryotic cells, with significant implications for pathogenesis of the strains encoding them. Meanwhile, T4SS is a more functionally diverse system, which is involved in not only effector translocation but also conjugation and DNA uptake/release. Development of control strategies against bacterial-mediated diseases requires genomic identification of the virulence arsenal of pathogenic bacteria, with T3SS, T4SS and T6SS being major determinants in this regard. Therefore, computational methods for systematic identification of these specialised machines are of particular interest. With the aim of facilitating this task, T346Hunter provides a user-friendly web-based tool for the prediction of T3SS, T4SS and T6SS clusters in newly sequenced bacterial genomes. After inspection of the available scientific literature, we constructed a database of hidden Markov model (HMM protein profiles and sequences representing the various components of T3SS, T4SS and T6SS. T346Hunter performs searches of such a database against user-supplied bacterial sequences and localises enriched regions in any of these three types of secretion systems. Moreover, through the T346Hunter server, users can visualise the predicted clusters obtained for approximately 1700 bacterial chromosomes and plasmids. T346Hunter offers great help to researchers in advancing their understanding of the biological mechanisms in which these sophisticated molecular machines are involved. T346Hunter is freely available at http://bacterial-virulence-factors.cbgp.upm.es/T346Hunter.

  11. DNA Delivery and Genomic Integration into Mammalian Target Cells through Type IV A and B Secretion Systems of Human Pathogens

    Directory of Open Access Journals (Sweden)

    Dolores L. Guzmán-Herrador

    2017-08-01

    Full Text Available We explore the potential of bacterial secretion systems as tools for genomic modification of human cells. We previously showed that foreign DNA can be introduced into human cells through the Type IV A secretion system of the human pathogen Bartonella henselae. Moreover, the DNA is delivered covalently attached to the conjugative relaxase TrwC, which promotes its integration into the recipient genome. In this work, we report that this tool can be adapted to other target cells by using different relaxases and secretion systems. The promiscuous relaxase MobA from plasmid RSF1010 can be used to deliver DNA into human cells with higher efficiency than TrwC. MobA also promotes DNA integration, albeit at lower rates than TrwC. Notably, we report that DNA transfer to human cells can also take place through the Type IV secretion system of two intracellular human pathogens, Legionella pneumophila and Coxiella burnetii, which code for a distantly related Dot/Icm Type IV B secretion system. This suggests that DNA transfer could be an intrinsic ability of this family of secretion systems, expanding the range of target human cells. Further analysis of the DNA transfer process showed that recruitment of MobA by Dot/Icm was dependent on the IcmSW chaperone, which may explain the higher DNA transfer rates obtained. Finally, we observed that the presence of MobA negatively affected the intracellular replication of C. burnetii, suggesting an interference with Dot/Icm translocation of virulence factors.

  12. Identification of a PVL-negative SCCmec-IVa sub-lineage of the methicillin-resistant Staphylococcus aureus CC80 lineage

    DEFF Research Database (Denmark)

    Edslev, Sofie Marie; Westh, Henrik Torkil; Andersen, Paal Skytt

    2018-01-01

    of the CC80 S. aureus lineage was conducted from whole-genome sequences of 217 isolates (23 MSSA and 194 MRSA) from 22 countries. All isolates were further genetically characterized in regard to resistance determinants and PVL carriage, and epidemiological data was obtained for selected isolates. RESULTS......OBJECTIVES: Community-acquired MRSA (CA-MRSA) isolates belonging to clonal complex 80 (CC80) are recognized as the European CA-MRSA. The prevailing European CA-MRSA clone carries a type IVc staphylococcal cassette chromosome mec (SCCmec) and expresses Panton-Valentine leukocidin (PVL). Recently......, a significant increase of PVL-negative CC80 MRSA has been observed in Denmark. The aim of this study was to examine the genetics and the epidemiology of these, and to compare them to the European CA-MRSA clone in order to understand the emergence of PVL-negative CC80 MRSA. METHODS: Phylogenetic analysis...

  13. Production of basement membrane laminin and type IV collagen by tumors of striated muscle: an immunohistochemical study of rhabdomyosarcomas of different histologic types and a benign vaginal rhabdomyoma.

    Science.gov (United States)

    Autio-Harmainen, H; Apaja-Sarkkinen, M; Martikainen, J; Taipale, A; Rapola, J

    1986-12-01

    Immunohistochemical methods were used to demonstrate the distribution of basement membrane laminin and type IV collagen in eight tumors derived from striated muscle (three botryoid, two alveolar, and two adult-type rhabdomyosarcomas; one benign vaginal rhabdomyoma). All of the tumors produced significant amounts of both basement membrane components. Stainings clearly revealed the alveolar nature of the rhabdomyosarcomas, with the alveolar spaces surrounded by distinct basement membranes. Different stages of cellular development were identified in the botryoid sarcomas, with the most immature cells of the cambium layer devoid of external basement membrane around the tumor cells, although the stroma contained finely dispersed basement membrane material and some cells contained intracytoplasmic laminin or type IV collagen, indicative of the synthesis of these proteins. The more mature cells, which had abundant granular cytoplasm, were enveloped by distinct basement membranes and seemed to have coalesced, forming structures resembling myotubes. The adult-type rhabdomyosarcomas were composed of large pleomorphic cells that were surrounded by basement membranes, either individually or in small groups. Some giant cells contained intracytoplasmic laminin. The vaginal rhabdomyoma was composed of round rhabdoblastic cells or elongated strap cells with cross-striations. Cells of both of these types were surrounded by thin but distinct basement membranes. The results suggest that demonstration of basement membranes would be helpful in the diagnosis of tumors derived from striated muscle. The findings concerning different stages of maturation of tumor cells are in accordance with previous in vitro observations of myoblastic cells.

  14. Spread of epidemic MRSA-ST5-IV clone encoding PVL as a major cause of community onset staphylococcal infections in Argentinean children.

    Directory of Open Access Journals (Sweden)

    Claudia Sola

    associated with a SCCmec type closely related to SCCmecIV(2B&5. CONCLUSIONS/SIGNIFICANCE: The dissemination of epidemic MRSA clone, ST5-IV-PVL(+ was the main cause of increasing staphylococcal community-onset infections in Argentinean children (2003-2008, conversely to other countries. The predominance of this clone, which has capacity to express the h-VISA phenotype, in healthcare-associated community-onset cases suggests that it has infiltrated into hospital-settings.

  15. Spread of Epidemic MRSA-ST5-IV Clone Encoding PVL as a Major Cause of Community Onset Staphylococcal Infections in Argentinean Children

    Science.gov (United States)

    Sola, Claudia; Egea, Ana L.; Moyano, Alejandro J.; Garnero, Analia; Kevric, Ines; Culasso, Catalina; Vindel, Ana; Lopardo, Horacio; Bocco, José L.

    2012-01-01

    and was associated with a SCCmec type closely related to SCCmecIV(2B&5). Conclusions/Significance The dissemination of epidemic MRSA clone, ST5-IV-PVL+ was the main cause of increasing staphylococcal community-onset infections in Argentinean children (2003–2008), conversely to other countries. The predominance of this clone, which has capacity to express the h-VISA phenotype, in healthcare-associated community-onset cases suggests that it has infiltrated into hospital-settings. PMID:22291965

  16. The extracellular matrix of Gadus morhua muscle contains types III, V, VI and IV collagens in addition to type I

    DEFF Research Database (Denmark)

    Brüggemann, Dagmar Adeline; Lawson, M.A.

    2005-01-01

    Confocal microscopy and immuno‐histochemistry were used to examine collagens in the extracellular matrix of cod Gadus morhua swimming muscle. In addition to the well known presence of type I fibrous collagen, types III and VI were also found in the myocommata and the endomysium. The beaded collagen...

  17. Loss of Col3a1, the gene for Ehlers-Danlos syndrome type IV, results in neocortical dyslamination.

    Directory of Open Access Journals (Sweden)

    Sung-Jin Jeong

    Full Text Available It has recently been discovered that Collagen III, the encoded protein of the type IV Ehlers-Danlos Syndrome (EDS gene, is one of the major constituents of the pial basement membrane (BM and serves as the ligand for GPR56. Mutations in GPR56 cause a severe human brain malformation called bilateral frontoparietal polymicrogyria, in which neurons transmigrate through the BM causing severe mental retardation and frequent seizures. To further characterize the brain phenotype of Col3a1 knockout mice, we performed a detailed histological analysis. We observed a cobblestone-like cortical malformation, with BM breakdown and marginal zone heterotopias in Col3a1⁻/⁻ mouse brains. Surprisingly, the pial BM appeared intact at early stages of development but starting as early as embryonic day (E 11.5, prominent BM defects were observed and accompanied by neuronal overmigration. Although collagen III is expressed in meningeal fibroblasts (MFs, Col3a1⁻/⁻ MFs present no obvious defects. Furthermore, the expression and posttranslational modification of α-dystroglycan was undisturbed in Col3a1⁻/⁻ mice. Based on the previous finding that mutations in COL3A1 cause type IV EDS, our study indicates a possible common pathological pathway linking connective tissue diseases and brain malformations.

  18. Herpes-simplex virus encephalitis is characterized by an early MMP-9 increase and collagen type IV degradation.

    Science.gov (United States)

    Sellner, Johann; Simon, Franziska; Meyding-Lamade, Uta; Leib, Stephen L

    2006-12-13

    Cerebrovascular complications including cerebral edema, raised intracranial pressure and hemorrhage contribute to the high mortality and morbidity of herpes-simplex virus encephalitis (HSE). We examined changes of collagen type IV, the major constituent of the neurovascular matrix, together with expression and localization of matrix-degrading enzymes during the development of acute HSE. In an experimental model of focal HSE, we found that early, symptomatic HSE (3 days after infection) and acute, fully developed HSE (7 days after infection) are associated with significantly raised levels of matrix-metalloproteinase-9 (MMP-9) (both PHSE and further expanded towards the perivascular space and adjacent tissue in acute HSE. Around the cerebral vasculature, we observed that MMP-9 activity was insufficiently counterbalanced by its endogenous tissue inhibitor of MMP (TIMP) TIMP-1, resulting in loss of collagen type IV. Our findings suggest that MMP-9 is involved in the evolution of HSE by causing damage to the cerebral vasculature. The degradation of the neurovascular matrix in HSE facilitates the development of cerebrovascular complications and may represent a target for novel adjuvant treatment strategies.

  19. A Comparative Structure/Function Analysis of Two Type IV Pilin DNA Receptors Defines a Novel Mode of DNA Binding.

    Science.gov (United States)

    Berry, Jamie-Lee; Xu, Yingqi; Ward, Philip N; Lea, Susan M; Matthews, Stephen J; Pelicic, Vladimir

    2016-06-07

    DNA transformation is a widespread process allowing bacteria to capture free DNA by using filamentous nano-machines composed of type IV pilins. These proteins can act as DNA receptors as demonstrated by the finding that Neisseria meningitidis ComP minor pilin has intrinsic DNA-binding ability. ComP binds DNA better when it contains the DNA-uptake sequence (DUS) motif abundant in this species genome, playing a role in its trademark ability to selectively take up its own DNA. Here, we report high-resolution structures for meningococcal ComP and Neisseria subflava ComPsub, which recognize different DUS motifs. We show that they are structurally identical type IV pilins that pack readily into filament models and display a unique DD region delimited by two disulfide bonds. Functional analysis of ComPsub defines a new mode of DNA binding involving the DD region, adapted for exported DNA receptors. Copyright © 2016 The Author(s). Published by Elsevier Ltd.. All rights reserved.

  20. Thermal Simulation Analysis of Microstructure and Hardness for CrMoV with PWHT in Type IV Region

    Science.gov (United States)

    Xu, Bin; Xue, Song; Cai, Yong; Niu, Jitai; Li, Liangchao

    2017-12-01

    Welded components of CrMoV steam pipe exhibit a pernicious form of type IV cracking after long-term service at elevated temperature. To investigate the cracking mechanism, the type IV microstructure and hardness were characterized after thermal simulation of post-weld heat treatment. Below 1098 K (825 °C), loss of carbon from the pearlite region was apparent, and the work zone exhibited a slightly lower hardness than the parent material because of a minor amount of austenite transformation. In addition, for peak temperatures above 1133 K (860 °C), additional transformation into austenite occurred and was followed by retransformation into ferrite upon further increasing the temperature. The pearlite formed at 1173 K to 1223 K (900 °C to 950 °C) resulted in an increase of the volume fraction of pearlite and microstructural refinement, which yielded a remarkable increase of hardness in the work zone. For the peak temperature of 1573 K (1300 °C), previous austenite grains were coarsened and alloy carbides were dissolved in the austenite, which significantly hardened the work zone.

  1. Porphyromonas gingivalis Induced Fragmentation of Type IV Collagen Through Macrophage-Activated MMP-9: (In Vitro Study of Collagenolytic Mechanism in Pathogenesis of Atherosclerotic Plaque Rupture

    Directory of Open Access Journals (Sweden)

    Siti Nurul Mubarokah

    2009-12-01

    Full Text Available BACKGROUND: Periodontitis is caused mostly by Porphyromonas gingivalis (P.gingivalis and it is related to acute coronary syndrome. P.gingivalis  readily invades blood circulation and potentially induces collagenolytic activity of inflammatory cells that results in collagen vascular degradation leading to atherosclerotic plague rupture (APR. APR is responsible for the occurence of fatal cardiovascular events such as acute myocardial infraction (AMI. AIMS: To show that P.gingivalis potentially induces fragmentation of the type IV vascular collagen due to macrophage-activated MMP-9. METHODS: The ability of P.gingivalis to induce the type IV collagen fragmentation, shown by digesting type IV collagen with the supernatant of monocyte-derived macrophage activated by exposure to P.gingivalis suspension for 18 hours, 37oC, 5% CO2. The type IV collagen fragments were analyzed by SDS-PAGE and confirmed by Western-blotting. Antibody of type IV collagen produced and confirmed by dot-blotting prior to its being used as primary antibody of Western-blotting. The existence of MMP-9 was detected by Dot-blot and Western-blot technique, while the MMP-9 activity was assessed by SDS-PAGE and zymograms. RESULTS: Our data showed that P.gingivalis induced macrophage to produce MMP-9 as one of collagenolytic components, and interaction with P.gingivalis proteases enhanced the proteolytic activity and resulted in degradation of type IV collagen with molecular weight of 88 kDa into two smaller fragments with molecular weight of 80 kDa and 60 kDa. CONCLUSIONS: P.gingivalis induced macrophage to activate its MMP-9 that led to fragmentation of vascular type IV collagen in the pathogenesis of atherosclerotic plaque rupture. KEYWORDS: P.gingivalis, macrophage, type IV collagen fragmentation, atherosclerotic plaque rupture, AMI.

  2. Homologies between the non-collagenous C-terminal (NC1) globular domains of the alpha 1 and alpha 2 subunits of type-IV collagen.

    Science.gov (United States)

    Kaytes, P S; Theriault, N Y; Vogeli, G

    1987-01-01

    The non-collagenous C-terminal globular domain (NC1) of type-IV collagen has the dual role of initiating triple-helix formation among the subunits and of crosslinking two collagen molecules during basement-membrane meshwork formation. By cloning a cDNA for the NC1 domain of the alpha 2(IV) collagen chain, we have found a high degree of homology (63% for nucleotides, 66% for amino acids) between the NC1 of the alpha 2 and alpha 1 chains of type-IV collagen. All cysteine residues are conserved. This high degree of homology is not found within the helical portion where the homology is 41% for amino acids (only 14% if the obligatory glycine is not used for this analysis). We propose that this high degree of homology within the non-collagenous domain indicates a close evolutionary relationship maintained by functional restraints between the two chains of type IV collagen.

  3. Diagnosing Mucopolysaccharidosis type IV a by the fluorometric assay of N-Acetylgalactosamine-6-sulfate sulfatase activity.

    Science.gov (United States)

    Shams, Sedigheh; Barazandeh Tehrani, Maliheh; Civallero, Gabriel; Minookherad, Koosha; Giugliani, Roberto; Setoodeh, Aria; Haghi Ashtiani, Mohammad Taghi

    2017-01-01

    Mucopolysaccharidosis type IVA, also known as Morquio A or MPS IV A, is an autosomal recessive disease caused by the deficiency of the lysosomal enzyme N-acetylgalactosamine-6-sulfate sulfatase (GALNS). The loss of GALNS activity leads to the impaired breakdown of glycosaminoglycans (GAGs) keratan sulfate and chondroitin-6-sulfate. The accumulation of GAGs results in multiple organ damage. The accurate and early diagnosis of this disorder helps enhance the effectiveness of the treatment. The present study uses a pre-designed protocol for testing GALNS activity in the leukocytes of Iranian patients with MPS IV A and their parents and compares it with healthy controls. Patients with MPS IVA previously diagnosed through the measurement of enzyme activity or genetic analysis entered the study. Leukocytes were obtained from the heparinized blood of the participants. The GALNS activity was measured by a fluorometric method using 4-methylumbelliferyl-β-D-galactoside-6-sulfate (4MU-G6S) as the substrate and proper buffer solutions and calibrators. The GALNS activity (nmol/17 h/mg protein) was reported as 0-7.4 in the MPSIV A patients, as 19.85-93.7 in their parents and as 38.4-164 in the healthy controls. Statistically significant differences were observed between the three groups in terms of enzyme activity. There were no significant differences in enzyme activity by age. The female subjects in both the patient and parents groups showed lower enzyme activity compared to the male subjects. The fluorometric method was validated for the measurement of GALNS activity in leukocyte samples and identifying Iranian patients with MPS IV A.

  4. Calcitonin Gene-Related Peptide Regulates Type IV Hypersensitivity through Dendritic Cell Functions: e86367

    National Research Council Canada - National Science Library

    Norihisa Mikami; Kaori Sueda; Yusuke Ogitani; Ippei Otani; Miku Takatsuji; Yasuko Wada; Keiko Watanabe; Rintaro Yoshikawa; Satoshi Nishioka; Nagisa Hashimoto; Yayoi Miyagi; So-ichiro Fukada; Hiroshi Yamamoto; Kazutake Tsujikawa

    2014-01-01

    .... In this study, we show that CGRP suppressed Th1 cell differentiation via inhibition of IL-12 production in DCs using an in vitro co-culture system and an in vivo ovalbumin-induced delayed-type hypersensitivity (DTH) model...

  5. Tissue-engineered cartilaginous constructs for the treatment of caprine cartilage defects, including distribution of laminin and type IV collagen.

    Science.gov (United States)

    Jeng, Lily; Hsu, Hu-Ping; Spector, Myron

    2013-10-01

    The purpose of this study was the immunohistochemical evaluation of (1) cartilage tissue-engineered constructs; and (2) the tissue filling cartilage defects in a goat model into which the constructs were implanted, particularly for the presence of the basement membrane molecules, laminin and type IV collagen. Basement membrane molecules are localized to the pericellular matrix in normal adult articular cartilage, but have not been examined in tissue-engineered constructs cultured in vitro or in tissue filling cartilage defects into which the constructs were implanted. Cartilaginous constructs were engineered in vitro using caprine chondrocyte-seeded type II collagen scaffolds. Autologous constructs were implanted into 4-mm-diameter defects created to the tidemark in the trochlear groove in the knee joints of skeletally mature goats. Eight weeks after implantation, the animals were sacrificed. Constructs underwent immunohistochemical and histomorphometric evaluation. Widespread staining for the two basement membrane molecules was observed throughout the extracellular matrix of in vitro and in vivo samples in a distribution unlike that previously reported for cartilage. At sacrifice, 70% of the defect site was filled with reparative tissue, which consisted largely of fibrous tissue and some fibrocartilage, with over 70% of the reparative tissue bonded to the adjacent host tissue. A novel finding of this study was the observation of laminin and type IV collagen in in vitro engineered cartilaginous constructs and in vivo cartilage repair samples from defects into which the constructs were implanted, as well as in normal caprine articular cartilage. Future work is needed to elucidate the role of basement membrane molecules during cartilage repair and regeneration.

  6. MANAGEMENT OF A GIANT TYPE IV A CHOLEDOCHAL CYST: CASE REPORT

    Directory of Open Access Journals (Sweden)

    Narendranath Nagoti

    2015-02-01

    Full Text Available Choledochal cysts are uncommon congenital anomalies of the biliary tree defined as abnormal, disproportionate, cystic dilatations of the biliary duct . (1 We report a 20 month old female child who presented with obstructive jaundice with recurrent episodes of cholangitis. She was diagnosed with a giant type IVA choledochal cyst with huge cystic dilatation of common bile duct (CBD and multiple cystic dilatations of the intrahepatic biliary channels. The patient experienced complete recovery after the cyst was excised using LIL L Y technique and biliary reconstruction was done using Roux - en - Y hepatico jejunostomy at the level of the hilum. Here we discuss the clinical features and management of this giant type IVA choledochal cyst

  7. Type IV hypersensitivity reactions following Dermabond adhesive utilization in knee surgery: A Report of Three Cases.

    Science.gov (United States)

    Yagnatovsky, Michelle; Pham, Hien; Rokito, Andrew; Jazrawi, Laith; Strauss, Eric

    2017-05-01

    ​We retrospectively reviewed the records of 3 patients (3 knees) with a delayed type hypersensitivity reaction following Dermabond exposure after an orthopaedic knee procedure. Delayed hypersensitivity reactions are mediated by CD4+ helper T cells. The use of skin adhesives in place of traditional sutures is increasing in popularity given Dermabond's potential benefits of decreased wound infection rate and better wound approximation. However, hypersensitivity reactions to the cyanoacrylate material in Dermabond have been described. Differentiating hypersensitivity reactions from post-operative infections is important as septic arthritis is a potentially devastating complication. This case series presents the challenge of properly diagnosing and managing hypersensitivity reactions. Consultation with allergists and dermatologists may be appropriate for ascertaining the nature of the surgical site complication and proper management. The recommended management of hypersensitivity-type reactions is a course of topical steroids and infection work up if needed.

  8. Treatment of Medial Malleolus or Pure Deltoid Ligament Injury in Patients with Supination-External Rotation Type IV Ankle Fractures.

    Science.gov (United States)

    Wang, Xu; Zhang, Chao; Yin, Jian-Wen; Wang, Chen; Huang, Jia-Zhang; Ma, Xin; Wang, Cheng-Wei; Wang, Xue

    2017-02-01

    To investigate the effect of internal fixation on postoperative ankle function in patients with supination-external rotation type IV ankle fractures, including medial malleolus fractures and deltoid ligament injury. Between January 2012 and June 2014, patients with medial structure injuries were enrolled in this study and assigned to the medial malleolus fracture group or the deltoid ligament group. The surgical procedures for the two groups were documented. The follow-up endpoint was the time point when the steel plate or screw was removed from the lateral ankle. The Olerud-Molander ankle scoring system was used to assess ankle function. A total of 84 patients with supination-external rotation type IV ankle fractures had complete medical records and were included in this study. The average age of the patients was 44.16 years (range, 15-75). The patient sample included 39 males and 45 females. Overall, 49 patients (19 males and 30 females) suffered a medial malleolus fracture. The average age of these patients was 40.20 years (range, 15-75). Patients with a posterior malleolar fracture fragment >25% of the articular surface accounted for 81.6% (40 patients) of these patients. Overall, 35 patients (20 males and 15 females) experienced a deltoid ligament injury. The average age of these patients was 44.21 years (range, 17-73). Patients with a posterior malleolar fracture fragment >25% of the articular surface accounted for 11.5% (four patients) of these patients. Open reduction was performed in patients with medial malleolus fractures, and two 4.0-mm cannulated screws were used to fixate the posterior malleolus and the medial malleolus. The suture-anchor technique was used to repair the ligaments in patients with deltoid ligament injuries. The follow-up endpoint was the time point when the steel plate and screws were removed from the lateral ankle in patients. The average follow-up period was 13.4 months (range, 11-17). The Olerud-Molander ankle scoring system was

  9. Identification of Major Sequence Types among Multidrug-Resistant Staphylococcus epidermidis Strains Isolated from Infected Eyes and Healthy Conjunctiva

    Science.gov (United States)

    Jena, Smrutiti; Panda, Sasmita; Nayak, Kinshuk C.; Singh, Durg V.

    2017-01-01

    We examined the presence of virulence and antibiotic resistance genes, SCCmec types and determined the genomic diversity among ocular S. epidermidis isolates (patients-23, healthy controls-29). PCR determined the presence of antibiotic resistance genes, virulence genes and SCCmec types among all isolates. MLST and PFGE determined the genomic relatedness among them. All isolates of S. epidermidis showed resistance to at least one class of antibiotics of which 48 isolates were multidrug resistant and carried ARGs. Thirty-five isolates were methicillin resistant and carried mecA gene. Majority of the isolates were resistant to fluoroquinolones and showed mutation in gyrA, parC, and parE genes, however, few isolates showed additional novel mutations in parC gene. Of the MRSE strains, 17 strains carried SCCmec type IV, four type V, two type II, and two UT4. Seven strains carried novel combination of ccr complex and SCCmercury element, not reported earlier. All the S. epidermidis strains harbored icaA and icaD genes, 47 carried ACME operon, and 50 contained IS256. A noteworthy finding was the presence of ST179 among 43% of infected eye isolates an observation rarely reported among S. epidermidis. PFGE and MLST analysis showed genomic diversity among them. Statistical analysis suggests that few healthy conjunctiva isolates had characteristics similar to infected eye isolates. S. epidermidis strains carrying mecA gene are multidrug resistant, virulent and diverse irrespective of sources of isolation. IS256 cannot be used as marker to differentiate isolates of infected eye from healthy conjunctiva. PMID:28824564

  10. Pseudomonas aeruginosa d-Arabinofuranose Biosynthetic Pathway and Its Role in Type IV Pilus Assembly*

    OpenAIRE

    Harvey, Hanjeong; Kus, Julianne V.; Tessier, Luc; Kelly, John; Burrows, Lori L.

    2011-01-01

    Pseudomonas aeruginosa strains PA7 and Pa5196 glycosylate their type IVa pilins with α1,5-linked d-arabinofuranose (d-Araf), a rare sugar configuration identical to that found in cell wall polymers of the Corynebacterineae. Despite this chemical identity, the pathway for biosynthesis of α1,5-d-Araf in Gram-negative bacteria is unknown. Bioinformatics analyses pointed to a cluster of seven P. aeruginosa genes, including homologues of the Mycobacterium tuberculosis genes Rv3806c, Rv3790, and Rv...

  11. Calcitonin gene-related peptide regulates type IV hypersensitivity through dendritic cell functions.

    Directory of Open Access Journals (Sweden)

    Norihisa Mikami

    Full Text Available Dendritic cells (DCs play essential roles in both innate and adaptive immune responses. In addition, mutual regulation of the nervous system and immune system is well studied. One of neuropeptides, calcitonin gene-related peptide (CGRP, is a potent regulator in immune responses; in particular, it has anti-inflammatory effects in innate immunity. For instance, a deficiency of the CGRP receptor component RAMP 1 (receptor activity-modifying protein 1 results in higher cytokine production in response to LPS (lipopolysaccharide. On the other hand, how CGRP affects DCs in adaptive immunity is largely unknown. In this study, we show that CGRP suppressed Th1 cell differentiation via inhibition of IL-12 production in DCs using an in vitro co-culture system and an in vivo ovalbumin-induced delayed-type hypersensitivity (DTH model. CGRP also down-regulated the expressions of chemokine receptor CCR2 and its ligands CCL2 and CCL12 in DCs. Intriguingly, the frequency of migrating CCR2(+ DCs in draining lymph nodes of RAMP1-deficient mice was higher after DTH immunization. Moreover, these CCR2(+ DCs highly expressed IL-12 and CD80, resulting in more effective induction of Th1 differentiation compared with CCR2(- DCs. These results indicate that CGRP regulates Th1 type reactions by regulating expression of cytokines, chemokines, and chemokine receptors in DCs.

  12. Intimal aortic sarcoma mimicking ruptured thoracoabdominal type IV aneurysm. a rare case report and review of the literature

    Directory of Open Access Journals (Sweden)

    Dedeilias Panagiotis

    2011-12-01

    Full Text Available Abstract Primary intimal aortic sarcoma represents a very rare and highly lethal medical entity. Diagnosis is made either by embolic events caused by the tumor or by surrounding tissue symptoms such as pain. Herein we report an extremely rare case of a 51-year-old man previously operated for ascending aortic aneurysm, who presented with clinical and radiological findings suggestive of a ruptured thoracoabdominal type IV aneurysm. The patient underwent radical resection of the aorta and surrounding tissue with placement of a composite 4-branched graft. The diagnosis was made by frozen section and regular histopathologic examination of the specimen and the patient received adjuvant chemotherapy. Nine months after surgery the patient is still alive and has no signs of recurrence. We review the literature and discuss the option of postoperative chemotherapy.

  13. Biphenyl derived oxovanadium(IV) and copper(II) salen-type complexes--structure and redox tuning.

    Science.gov (United States)

    Plitt, Patrick; Pritzkow, Hans; Oeser, Thomas; Kraemer, Roland

    2005-05-01

    A series of vanadyl(IV) salen (N,N'-bis(salicylidene)ethylenediaminato)-type complexes (1-4) bearing phenyl or 2-hydroxyphenyl moieties have been prepared and characterized by means of mass spectrometry, infra-red, electron paramagnetic resonance (EPR), UV/Vis spectroscopy, cyclovoltammetry and X-ray crystallography. Their structures have been compared to their copper(II) analogs 5-8. Hydrogen intralinkages have been observed in the crystal structure of 5. The pendant hydroxy groups fine-tune the redox properties of the complexes. The catalytic activity in the oxygenation of ethyl phenyl sulfide to the corresponding sulfoxide was investigated. Results indicate that complex 1 bearing hydroxyphenyl subunits and a phenylene bridge is the most selective under these reaction conditions, with the smallest amount of the over-oxidized product, sulfone.

  14. [Total hip arthroplasty for crowe type IV developmental dysplasia of the hip with S-ROM prosthesis].

    Science.gov (United States)

    Sun, Qi-Cai; Wang, Xiang-Hua; Song, Bai-Shan; Zhu, Fang-Bing; Yan, Shi-gui

    2013-02-01

    To develop the techniques of total hip arthroplasty(THA) for Crowe type IV developmental dysplasia of the hip (DDH) with S-ROM prosthesis,and to assess its clinical results. From October 2000 to October 2011,30 patients (36 hips) with Crowe type IV DDH underwent THA,including 6 patients with bilateral hip involved and 24 patients with unilateral. S-ROM prosthesis was adopted together with subtrochanteric transverse osteotomy. All the cementless acetabular cups were placed at the original anatomic location. The threaded cups were put in or near the level of the true acetabulum in all patients. Full coating stems were used in femoral side. All the patients were evaluated by using the Modified Harris Hip Score. Radiographic evaluations were made preoperatively and during follow-up. Two patients lost of follow-up. Twenty-seven patients with 32 hips were followed up,and the average duration was 48 months (ranging from 7 to 84 months). There was 1 patient with bilateral THA died from hemorrhagic shock. Two patients could walk freely with the visible fracture lines at 12th and 18th months postoperatively. There were no complications such as infection or nerve injuries. Modified Harris Hip Score improved from preoperative 41.7+/-3.7 to postoperative 89.1+/-2.9. There was no acetabular or femoral component revision because of mal-position or loosening of the prostheses in all patients. Postoperative X-ray showed that all the prostheses in place,good integration between acetabular cups,femoral prosthesis and host bone without loosening. All bone grafts were integrated. All the hips acquired union of osteotomy and bone in-growth. None of the patients had radiographic evidence of aseptic loosening of prosthesis. For the complex DDH, follow methods should be used to improve therapeutic effects:good exposure of the true acetabulum,deepen acetabulum, femoral shortening, oblique osteotomy, using the S-ROM prosthesis.

  15. Photon storage in ¿-type optically dense atomic media. IV. Optimal control using gradient ascent

    DEFF Research Database (Denmark)

    Gorshkov, Alexey V.; Calarco, Tomasso; Lukin, Mikhail D.

    2008-01-01

    We use the numerical gradient ascent method from optimal control theory to extend efficient photon storage in -type media to previouslyinaccessible regimes and to provide simple intuitive explanations for our optimization techniques. In particular, by using gradient ascent to shape classical....... We also demonstrate that the often discussed connection between time reversal andoptimality in photon storage follows naturally from gradient ascent. Finally, we discuss the optimization of controlled reversible inhomogeneous broadening....... control pulses used to mediate photon storage, we open up the possibility of high efficiency photon storage in thenonadiabatic limit, in which analytical solutions to the equations of motion do not exist. This control shaping technique enables an order-of-magnitude increase in the bandwidth of the memory...

  16. Genotype–Phenotype Correlations in Pathology Caused by Collagen Type IV alpha 1 and 2 Mutations

    Science.gov (United States)

    Jeanne, Marion; Gould, Douglas B

    2017-01-01

    COL4A1 and COL4A2 are extracellular matrix proteins that form heterotrimers and are present in nearly all basement membranes in every organ. In the past decade, COL4A1 and COL4A2 mutations have been identified to cause a multi-system disorder for which penetrance and severity of constituent phenotypes can greatly vary. Here, we compare the outcomes of more than 100 mutations identified in patients and data from a murine allelic series to explore the presence of genotype-phenotype correlations – many of which are shared among other types of collagen. We find that there is a frequency bias for COL4A1 over COL4A2 mutations and that glycine (Gly) substitutions within the triple helical domain are the most common class of mutations. Glycine is most often replaced by a charged amino acid, however the position of the mutation, and not the properties of the substituting amino acid, appear to have a greater influence on disease severity. Moreover, the impact of position is not straightforward. Observations from a murine allelic series suggest that mutations in the NC1 domain may result in relatively mild phenotypes via a ‘quantitative’ mechanism similar to other types of collagens, however, this effect was not apparent in human reports. Importantly, other position-dependent effects had differential impacts depending on the phenotype of interest. For example, the severity of cerebrovascular disease correlated with an amino-to-carboxy severity gradient for triple-helical glycine substitutions whereas the penetrance and severity of myopathy and nephropathy appear to involve a functional sub-domain(s). Greater understanding of genotype-phenotype correlations and the interaction of consequences of different mutations will be important for patient prognosis and care and for developing mechanism-based therapeutics to treat individual components of this emerging syndrome. PMID:27794444

  17. Surgical pitfalls in patients with Ehlers–Danlos type IV: A case of spontaneous sigmoid perforation in a 17-year-old male

    Directory of Open Access Journals (Sweden)

    Kai Lyn Ng

    2011-07-01

    Full Text Available Ehlers–Danlos syndrome (EDS is a group of well described connective tissue disorders in which collagen production is impaired. The surgical management of affected individuals remains challenging, with no general consensus. We report a case of spontaneous sigmoid perforation in a 17-year-old Eurasian male, in whom we subsequently established the diagnosis of EDS type IV (EDS-IV. We review the literature to discuss the clinical features and diagnosis, and the recommended therapeutic management.

  18. Modified Phemister procedure for the surgical treatment of Rockwood types III, IV, V acute acromioclavicular joint dislocation.

    Science.gov (United States)

    Verdano, M A; Pellegrini, A; Zanelli, M; Paterlini, M; Ceccarelli, F

    2012-12-01

    Acromioclavicular (AC) joint dislocations are common in young, active patients. In case of surgical indications, Rockwood type IV, type V and selected type III dislocation, we use modified Phemister procedure. At mid-term follow-up, by an average of 35.1 months, we reassessed the records of 14 patients surgically treated for acute AC dislocation from February 2007 to November 2010. In patients with a diagnosis of grade III lesion, indication for surgery was given on the basis of the patient's functional demand. Full recovery was obtained on average 3 months after surgery. Constant Score accounted for 92.7 points, mean Disabilities of the Arm, Shoulder and Hand Score for 3.2 points, and mean Simple Shoulder Test Score for 11.4 points. X-ray findings were partial loss of reduction (9 cases), subclinic re-dislocation (2 cases), calcification (5 cases) and post-traumatic arthritis (2 cases). Modified Phemister is a reliable technique, technically easy, a low learning curve procedure and cheap with low hardware's costs.

  19. SVBR-100 module-type fast reactor of the IV generation for regional power industry

    Science.gov (United States)

    Zrodnikov, A. V.; Toshinsky, G. I.; Komlev, O. G.; Stepanov, V. S.; Klimov, N. N.

    2011-08-01

    In the report the following is presented: basic conceptual provisions of the innovative nuclear power technology (NPT) based on modular fast reactors (FR) SVBR-100, summarized results of calculations of the reactor, analysis of the opportunities of multi-purpose application of such reactor facilities (RF) including export potentials with due account of nonproliferation requirements. The most important features of the proposed NPT analyzed in the report are as follows: (1) integral (monoblock) arrangement of the primary circuit equipment with entire elimination of the primary circuit pipelines and valves that considerably reduces the construction and assembly works period and coupling with high boiling point of lead-bismuth coolant (LBC) deterministically eliminates accidents of the LOCA type, (2) option for 100 MWe power and dimensions of the reactor provide: on the one hand, an opportunity to transport the reactor monoblock in factory-readiness by railway as well as other kinds of transport, on the other hand, core breeding ratio (CBR) exceeds 1 while MOX-fuel is used. The preferable area of application of RF SVBR-100 is regional and small power requiring power-units of electric power in a range of (100-600) MW, which could be used for cogeneration-based district heating while locating them nearby cities as well as for generation of electric power in a mode of load tracking in the regions with low network systems.

  20. Experimental holographic movie IV: the projection-type display system using a retro-directive screen

    Science.gov (United States)

    Higuchi, Kazuhito; Ishii, Ken'ichiro; Ishikawa, Jun; Hiyama, Shigeo

    1995-04-01

    Holographic movies can be seen as a tool to estimate the picture quality of moving holographic images as a step towards holographic television. The authors have previously developed three versions of an experimental holographic movie system, and this paper is a report on an improved version 4 of the system. The new version features a newly-developed projection-type display with a retro-directive beaded-screen, and an automatic film driver unit which moves perforated 35 mm holographic film intermittently with a shutter. A twin diamond-shaped hologram format, which was developed in the earlier version 2, is adopted for the films. The films comprise a series of reconstructed moving holographic images with minimal blurring. The optical arrangement and structure of the version 4 system enable the viewers to watch the film images in an open space, which in turn relieves them of the psychological pressure they felt with the previous three versions, when they had to squint into a narrow window built into a wall on the side of the device.

  1. [Non reported spa types in our country in Staphylococcus aureus from adult patients of a school hospital, Santa Fe - Argentina].

    Science.gov (United States)

    Tomatis, Carla; Baroni, María R; Mendosa, María A; Nagel, Alicia; Mollerach, Analia; Alvarez, Claudia; Zurbriggen, Maria Laura; Cristobal, Sabrina; Segovia, Glenda; Méndez, Emilce de Los A

    2018-01-11

    Staphylococcus aureus is a pathogen associated a different kind of infection. Molecular markers are useful tools to study microbial epidemiology. Twenty two methicillin-resistant S. aureus (MRSA) and 23 methicillin-susceptible S. aureus (MSSA) were studied by mecA gene, SCCmec cassette, Panton Valentine leucocidin (PVL) and spa polymorphism. The clinical data patients were analyzed. MSSA was prevalent in samples different from skin and soft tissue (SST) and in hospitalized patients, whereas MRSA in SST. SCCmec type IV was predominant, followed by type I. Low presence of PVL was found. In MRSA 11 different types of spa were detected, t019 was the most frequent and associated with outpatient, 17 types were found in MSSA and t189 was prevalent. spa t002 was present in MSSA and MRSA. We found 11 types of spa not reported in our country. Copyright © 2017 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  2. [Becker V-shaped lateral rotation osteotomy in total hip arthroplasty for Crowe type IV development dislocation of hip].

    Science.gov (United States)

    Li, Yongwang; Ma, Wenhai; Sun, Junying; Song, Xingjian; An, Ming; Zhang, Qian

    2013-09-01

    To summarize the surgical technique and the effectiveness of Becker V-shaped lateral rotation osteotomy in total hip arthroplasty (THA) for Crowe type IV development dislocation of the hip (DDH). Between January 2000 and December 2009, 18 patients (22 hips) with Crowe type IV DDH underwent THA. There were 3 males and 15 females with an average age of 54 years (range, 41-75 years). The unilateral hip was involved in 14 cases and bilateral hips in 4 cases. All patients had over anteversion of the femoral neck, with the acetabular anteversion angle of (21.28 +/- 4.87) degrees, the femoral neck anteversion angle of (59.06 +/- 1.44) degrees, and combined anteversion angle of (80.33 +/- 1.55) degrees. All the patients had limb-length discrepancy, ranged from 1.0 to 3.5 cm (mean, 2.5 cm). Before operation, gluteus medius muscle strength was grade 2 in 17 hips and grade 3 in 5 hips; severe or moderate claudication was observed in 13 and 5 patients, respectively. Trendelenburg sign was positive in all patients. Preoperative Harris score was 30.00 +/- 6.32. Cementless prosthesis was used. Becker V-shaped lateral rotation osteotomy and subtrochanteric shortening with overlapping femoral resection were performed, and proximal femoral shaft splitting was performed on 21 hips having narrow bone marrow cavity. All the cases achieved primary healing of incision. No complication of anterior dislocation, deep infection, nerve traction injury, or femoral uncontrolled fracture occurred. All the cases were followed up 3-12 years (mean, 8 years). Postoperative X-ray films showed that the initial fixation result of femoral prosthesis was excellent in 18 hips and good in 4 hips. Bone healing of osteotomy stump was obtained at 3-6 months (mean, 5 months) after operation. Affected limb prolonged for 2.5-3.5 cm (mean, 3.0 cm) at 1 year after operation; limb-length discrepancy was 0.5-1.5 cm (mean, 1.0 cm). The gluteus medius muscle strength was restored to grade 4 in 5 hips and grade 5 in 17

  3. Isolation of coagulase-positive staphylococci from bitches' colostrum and milk and genetic typing of methicillin-resistant Staphylococcus pseudintermedius strains.

    Science.gov (United States)

    Rota, Ada; Corrò, Michela; Drigo, Ilenia; Bortolami, Alessio; Börjesson, Stefan

    2015-07-23

    Among the coagulase-positive, potentially pathogenic staphylococci, Staphylococcus pseudintermedius has been frequently isolated from bitches' milk. This organism colonizes the mammary gland or causes infection, while S. aureus has been only occasionally reported. The objective of this study was to investigate the occurrence and persistence of coagulase-positive staphylococci in the colostrum and milk of postpartum bitches, either treated or untreated with antimicrobials, and to assess the incidence, antibiotic resistance profile and genetic type of the methicillin-resistant strains. On postpartum D1, D7 and D15, drops of secretion were collected from the mammary glands of 27 postpartum bitches, nine of which were treated with antimicrobials. Coagulase-positive staphylococci were identified, antimicrobial susceptibility and the presence of mecA were tested and the genetic profile of methicillin-resistant strains was assessed. Staphylococcus pseudintermedius was the only coagulase-positive staphylococcus isolated, and its presence was detected in 21 out of 27 bitches and in 66 out of 145 swabs. In a single bitch, it caused puerperal mastitis. In untreated bitches, the frequency of isolation was lower in colostrum than in milk. All of the isolates except one were resistant to at least three antimicrobial classes, while 14 out of 66 S. pseudintermedius strains were methicillin-resistant mecA positive (MRSP) and were isolated from eight bitches housed in the same breeding kennel. A significant association was found between antimicrobial treatment and the presence of MRSP. Six of the 12 typed isolates belonged to spa-type t02 carrying SCCmec II/III, and another six were non-typeable with spa carrying SCCmec IV. The t02-SCCmec II/III isolates were sequence type (ST) 71; four NT-SCCmec IV isolates were ST258 and two were ST369. PFGE showed that isolates from the same dog had identical band patterns, while isolates from different dogs had unique band patterns. MRSP strains

  4. Arctigenin, a phenylpropanoid dibenzylbutyrolactone lignan, inhibits type I-IV allergic inflammation and pro-inflammatory enzymes.

    Science.gov (United States)

    Lee, Ji Yun; Kim, Chang Jong

    2010-06-01

    We previously reported that arctigenin, a phenylpropanoid dibenzylbutyrolactone lignan isolated from Forsythia koreana, exhibits anti-inflammatory, antioxidant, and analgesic effects in animal models. In addition, arctigenin inhibited eosinophil peroxidase and activated myeloperoxidase in inflamed tissues. In this study, we tested the effects of arctigenin on type I-IV allergic inflammation and pro-inflammatory enzymes in vitro and in vivo. Arctigenin significantly inhibited the heterologous passive cutaneous anaphylaxis induced by ovalbumin in mice at 15 mg/kg, p.o., and compound 48/80-induced histamine release from rat peritoneal mast cells at 10 microM. Arctigenin (15 mg/kg, p.o.) significantly inhibited reversed cutaneous anaphylaxis. Further, arctigenin (15 mg/kg, p.o.) significantly inhibited the Arthus reaction to sheep's red blood cells, decreasing the hemolysis titer, the hemagglutination titer, and the plaque-forming cell number for SRBCs. In addition, arctigenin significantly inhibited delayed type hypersensitivity at 15 mg/kg, p.o. and the formation of rosette-forming cells at 45 mg/kg, p.o. Contact dermatitis induced by picrylchloride and dinitrofluorobenzene was significantly (p arctigenin (0.3 mg/ear). Furthermore, arctigenin dose-dependently inhibited pro-inflammatory enzymes, such as cyclooxygenase-1 and 2, 5-lipoxygenase, phospholipase A2, and phosphodiesterase. Our results show that arctigenin significantly inhibited B- and T-cell mediated allergic inflammation as well as pro-inflammatory enzymes.

  5. Fusion of lysosomes with secretory organelles leads to uncontrolled exocytosis in the lysosomal storage disease mucolipidosis type IV.

    Science.gov (United States)

    Park, Soonhong; Ahuja, Malini; Kim, Min Seuk; Brailoiu, G Cristina; Jha, Archana; Zeng, Mei; Baydyuk, Maryna; Wu, Ling-Gang; Wassif, Christopher A; Porter, Forbes D; Zerfas, Patricia M; Eckhaus, Michael A; Brailoiu, Eugen; Shin, Dong Min; Muallem, Shmuel

    2016-02-01

    Mutations in TRPML1 cause the lysosomal storage disease mucolipidosis type IV (MLIV). The role of TRPML1 in cell function and how the mutations cause the disease are not well understood. Most studies focus on the role of TRPML1 in constitutive membrane trafficking to and from the lysosomes. However, this cannot explain impaired neuromuscular and secretory cells' functions that mediate regulated exocytosis. Here, we analyzed several forms of regulated exocytosis in a mouse model of MLIV and, opposite to expectations, we found enhanced exocytosis in secretory glands due to enlargement of secretory granules in part due to fusion with lysosomes. Preliminary exploration of synaptic vesicle size, spontaneous mEPSCs, and glutamate secretion in neurons provided further evidence for enhanced exocytosis that was rescued by re-expression of TRPML1 in neurons. These features were not observed in Niemann-Pick type C1. These findings suggest that TRPML1 may guard against pathological fusion of lysosomes with secretory organelles and suggest a new approach toward developing treatment for MLIV. © 2015 The Authors.

  6. Caught in the act: the dialogue between bacteriophage R17 and the type IV secretion machine of plasmid R1.

    Science.gov (United States)

    Berry, Trista M; Christie, Peter J

    2011-12-01

    Bacteria communicate with each other through contact-independent and -dependent signalling mechanisms. Sensory perception of both types of signals is needed for conjugative transfer of mobile DNA elements via type IV secretion systems (T4SSs) to bacterial or eukaryotic target cells. While the regulatory circuitries coupling extracellular quorum and environmental signals to transcription of T4SS genes are increasingly understood, it remains fundamentally unknown how a potential recipient cell stimulates donor conjugative DNA transfer upon contact. In this issue, Lang et al. (2011) report use of the male-specific bacteriophage R17, a phage that binds conjugative pili elaborated by IncF plasmid R1, to define requirements for phage-contact-mediated T4SS activation and phage penetration. They report that R17 penetrates only through T4SS channels engaged for delivery of their plasmid cargo to recipient cells. Engagement requires docking of catalytically active relaxase TraI bound at oriT with the TraD substrate receptor (also termed the T4CP). The data, together with recent ultrastructural and biochemical findings, support an intriguing new model that the T4CP cumulatively senses an intracellular signal (substrate docking) and an extracellular signal (pilus bound by phage or a recipient cell) to co-ordinate a late stage morphogenetic or gating reaction that enables bidirectional transmission of nucleoprotein substrates through the T4SS. © 2011 Blackwell Publishing Ltd.

  7. A novel relaxase homologue is involved in chromosomal DNA processing for type IV secretion in Neisseria gonorrhoeae.

    Science.gov (United States)

    Salgado-Pabón, Wilmara; Jain, Samta; Turner, Nicholas; van der Does, Chris; Dillard, Joseph P

    2007-11-01

    The Neisseria gonorrhoeae type IV secretion system secretes chromosomal DNA that acts in natural transformation. To examine the mechanism of DNA processing for secretion, we made mutations in the putative relaxase gene traI and used nucleases to characterize the secreted DNA. The nuclease experiments demonstrated that the secreted DNA is single-stranded and blocked at the 5' end. Mutation of traI identified Tyr93 as required for DNA secretion, while substitution of Tyr201 resulted in intermediate levels of DNA secretion. TraI exhibits features of relaxases, but also has features that are absent in previously characterized relaxases, including an HD phosphohydrolase domain and an N-terminal hydrophobic region. The HD domain residue Asp120 was required for wild-type levels of DNA secretion. Subcellular localization studies demonstrated that the TraI N-terminal region promotes membrane interaction. We propose that Tyr93 initiates DNA processing and Tyr201 is required for termination or acts in DNA binding. Disruption of an inverted-repeat sequence eliminated DNA secretion, suggesting that this sequence may serve as the origin of transfer for chromosomal DNA secretion. The TraI domain architecture, although not previously described, is present in 53 uncharacterized proteins, suggesting that the mechanism of TraI function is a widespread process for DNA donation.

  8. Type IV collagen α1-chain noncollagenous domain blocks MMP-2 activation both in-vitro and in-vivo.

    Science.gov (United States)

    Sudhakar, Yakkanti Akul; Verma, Raj Kumar; Pawar, Smita C

    2014-03-26

    α1(IV)NC1 inhibits angiogenesis by regulating MAPK activation, this biological function was partly attributed α1(IV)NC1 binding to α1β1-integrin. However, its potent antiangiogenic activity and the molecular targets of α1(IV)NC1 has not been investigated. In the present study, the regulation of MMP-2 activation by α1(IV)NC1 was evaluated. α1β1-integrin which is required for inhibition of angiogenesis is not playing a role in cellular invasion and inhibition of MMP-2 activation by α1(IV)NC1. We found that α1(IV)NC1 binds the CBD of MMP-2 and forming a stable complex that prevents activation of MMP-2. The antiangiogenic activity of α1(IV)NC1 is mediated, in part, by this binding activity. In addition, up-regulation of TIMP-2 by α1(IV)NC1 led to saturation of MT1-MMP binding sites, which in turn led to inhibition of MMP-2 activation. In-vivo studies using α1-integrin null-mice treated with higher doses of α1(IV)NC1 showed integrin independent inhibition of tumor growth and active-MMP-2, without affecting MMP-9, MMP-7 and angiostatin.

  9. Lack of association of the HMGA1 IVS5-13insC variant with type 2 diabetes in an ethnically diverse hypertensive case control cohort

    Directory of Open Access Journals (Sweden)

    Karnes Jason H

    2013-01-01

    Full Text Available Abstract Background Recently, the high-mobility group A1 gene (HMGA1 variant IVS5-13insC has been associated with type 2 diabetes, but reported associations are inconsistent and data are lacking in Hispanic and African American populations. We sought to investigate the HMGA1-diabetes association and to characterize IVS5-13insC allele frequencies and linkage disequilibrium (LD in 3,070 Caucasian, Hispanic, and African American patients from the INternational VErapamil SR-Trandolapril STudy (INVEST. Methods INVEST was a randomized, multicenter trial comparing two antihypertensive treatment strategies in an ethnically diverse cohort of hypertensive, coronary artery disease patients. Controls, who were diabetes-free throughout the study, and type 2 diabetes cases, either prevalent or incident, were genotyped for IVS5-13insC using Taqman®, confirmed with Pyrosequencing and Sanger sequencing. For LD analysis, genotyping for eight additional HMGA1 single nucleotide polymorphisms (SNPs was performed using the Illumina® HumanCVD BeadChip. We used logistic regression to test association of the HMGA1 IVS5-13insC and diabetes, adjusted for age, gender, body mass index, and percentage European, African, and Native American ancestry. Results We observed IVS5-13insC minor allele frequencies consistent with previous literature in Caucasians and African Americans (0.03 in cases and 0.04 in controls for both race/ethnic groups, and higher frequencies in Hispanics (0.07 in cases and 0.07 in controls. The IVS5-13insC was not associated with type 2 diabetes overall (odds ratio 0.98 [0.76-1.26], p=0.88 or in any race/ethnic group. Pairwise LD (r2 of IVS5-13insC and rs9394200, a SNP previously used as a tag SNP for IVS5-13insC, was low (r2=0.47 in Caucasians, r2=0.25 in Hispanics, and r2=0.06 in African Americans. Furthermore, in silico analysis suggested a lack of functional consequences for the IVS5-13insC variant. Conclusions Our results suggest that IVS5-13ins

  10. A simplified multiplex PCR assay for fast and easy discrimination of globally distributed staphylococcal cassette chromosome mec types in meticillin-resistant Staphylococcus aureus.

    Science.gov (United States)

    Ghaznavi-Rad, Ehsanollah; Nor Shamsudin, Mariana; Sekawi, Zamberi; van Belkum, Alex; Neela, Vasanthakumari

    2010-10-01

    A multiplex PCR assay was developed for the identification of major types and subtypes of staphylococcal cassette chromosome mec (SCCmec) in meticillin-resistant Staphylococcus aureus (MRSA) strains. The method uses a novel 9 valent multiplex PCR plus two primer pairs for S. aureus identification and detection of meticillin resistance. All 389 clinical MRSA isolates from Malaysia and 18 European isolates from the Harmony collection harbouring different SCCmec types that we tested were correctly characterized by our PCR assay. SCCmec type III and V were by far the most common types among both hospital- and community-acquired Malaysian MRSA isolates, with an apparent emergence of MRSA harbouring the IVh type.

  11. Immunohistochemical localization of collagenous components in healthy periodontal tissues of the rat and marmoset (Callithrix jacchus). II. Distribution of collagen types IV, V and VI.

    Science.gov (United States)

    Romanos, G E; Schröter-Kermani, C; Hinz, N; Wachtel, H C; Bernimoulin, J P

    1991-07-01

    The immunohistochemical distribution of collagen types IV, V and VI has been demonstrated in healthy periodontal tissues of rats and marmosets following decalcification of the maxillae and mandibulae in 0.2 N HCl. An intense fluorescence with anti-collagen type IV antibodies was demonstrated in the basement membranes of the epithelium and of the blood vessels and nerves. In the alveolar bone stroma and in the periodontal ligament (PL) collagen type IV was present only in the basal membranes of the blood vessels and nerves. In comparison, collagen type V was observed in a fibrillar pattern in the gingival connective tissue, as well as the PL. In the PL, type V collagenous fibers demonstrated a parallel distribution with stronger fluorescence near the cementum surface. Collagen type VI could be demonstrated in fine fibers present in the gingival connective tissue and the PL. Blood vessels and nerves were not stained in the marmoset, but were in the rat, where a localization of collagen type VI was demonstrated in these areas. Alveolar bone and cementum, as well as the Sharpey's fibers embedded in these tissues, were not stained with antibodies against collagen type V and type VI, but a pericellular localization of these collagenous components could be observed. Collectively, these results provide basic information on the relative distribution of different collagen types in normal tissues of rats and marmosets that will be required for future studies on the effects of pathological, reparative and regenerative processes.

  12. A selective stepwise heme oxygenase model system: an iron(IV)-oxo porphyrin π-cation radical leads to a verdoheme-type compound via an isoporphyrin intermediate.

    Science.gov (United States)

    Garcia-Bosch, Isaac; Sharma, Savita K; Karlin, Kenneth D

    2013-11-06

    The selective oxidation of the α-position of two heme-Fe(III) tetraarylporphryinate complexes occurs when water(hydroxide) attacks their oxidized Cmpd I-type equivalents, high-valent Fe(IV)═O π-cation radical species ((P(+•))Fe(IV)═O). Stepwise intermediate formation occurs, as detected by UV-vis spectroscopic monitoring or mass spectrometric interrogation, being iron(III) isoporphyrins, iron(III) benzoyl-biliverdins, and the final verdoheme-like products. Heme oxygenase (HO) enzymes could proceed through heterolytic cleavage of an iron(III)-hydroperoxo intermediate to form a transient Cmpd I-type species.

  13. Y-configured metallic stent combined with 125 I seed strands cavity brachytherapy for a patient with type IV Klatskin tumor

    Directory of Open Access Journals (Sweden)

    Jiao Dechao

    2016-08-01

    Full Text Available We report a case in an inoperable patient with type IV Klatskin tumor treated by the use of a novel, two piece, Y-configured self-expandable metallic stent (SEMS combined with two 125 I seed strands via bilateral approach. The placement of the Y-shaped SEMS was successful and resulted in adequate biliary drainage. After 2 months of intraluminal brachytherapy (ILBT, both 125 I seed strands and temporary drainage catheter were removed after patency of the expanded stents was confirmed by the cholangiogram. This technique was feasible and could be considered for the treatment of patients with Bismuth type IV Klatskin tumors.

  14. Type-IVC Secretion System: A Novel Subclass of Type IV Secretion System (T4SS) Common Existing in Gram-Positive Genus Streptococcus

    Science.gov (United States)

    Chen, Chen; Gao, George F.

    2012-01-01

    A growing number of pathogens are being found to possess specialized secretion systems which they use in various ways to subvert host defenses. Type IV secretion system (T4SS) is one of versatile secretion systems essential for the virulence and even survival of some bacteria species, and they enable the secretion of protein and DNA substrates across the cell envelope. T4SS was once believed to be present only in Gram-negative bacteria. In this study, we present evidence of a new subclass of T4SS, Type-IVC secretion system and indicate its common existence in the Gram-positive bacterial genus Streptococcus. We further identified that VirB1, VirB4, VirB6 and VirD4 are the minimal key components of this system. Using genome comparisons and evolutionary relationship analysis, we proposed that Type-IVC secretion system is movable via transposon factors and mediates the conjugative transfer of DNA, enhances bacterial pathogenicity, and could cause large-scale outbreaks of infections in humans. PMID:23056296

  15. Pea Broth Enhances the Biocontrol Efficacy of Lysobacter capsici AZ78 by Triggering Cell Motility Associated with Biogenesis of Type IV Pilus.

    Science.gov (United States)

    Tomada, Selena; Puopolo, Gerardo; Perazzolli, Michele; Musetti, Rita; Loi, Nazia; Pertot, Ilaria

    2016-01-01

    Bacterial cells can display different types of motility, due to the presence of external appendages such as flagella and type IV pili. To date, little information on the mechanisms involved in the motility of the Lysobacter species has been available. Recently, L. capsici AZ78, a biocontrol agent of phytopathogenic oomycetes, showed the ability to move on jellified pea broth. Pea broth medium improved also the biocontrol activity of L. capsici AZ78 against Plasmopara viticola under greenhouse conditions. Noteworthy, the quantity of pea residues remaining on grapevine leaves fostered cell motility in L. capsici AZ78. Based on these results, this unusual motility related to the composition of the growth medium was investigated in bacterial strains belonging to several Lysobacter species. The six L. capsici strains tested developed dendrite-like colonies when grown on jellified pea broth, while the development of dendrite-like colonies was not recorded in the media commonly used in motility assays. To determine the presence of genes responsible for biogenesis of the flagellum and type IV pili, the genome of L. capsici AZ78 was mined. Genes encoding structural components and regulatory factors of type IV pili were upregulated in L. capsici AZ78 cells grown on the above-mentioned medium, as compared with the other tested media. These results provide new insight into the motility mechanism of L. capsici members and the role of type IV pili and pea compounds on the epiphytic fitness and biocontrol features of L. capsici AZ78.

  16. Pea broth enhances the biocontrol efficacy of Lysobacter capsici AZ78 by triggering cell motility associated with biogenesis of type IV pilus

    Directory of Open Access Journals (Sweden)

    Selena Tomada

    2016-07-01

    Full Text Available Bacterial cells can display different types of motility, due to the presence of external appendages such as flagella and type IV pili. To date, little information on the mechanisms involved in the motility of the Lysobacter species has been available. Recently, L. capsici AZ78, a biocontrol agent of phytopathogenic oomycetes, showed the ability to move on jellified pea broth. Pea broth medium improved also the biocontrol activity of L. capsici AZ78 against Plasmopara viticola under greenhouse conditions. Noteworthy, the quantity of pea residues remaining on grapevine leaves fostered cell motility in L. capsici AZ78. Based on these results, this unusual motility related to the composition of the growth medium was investigated in bacterial strains belonging to several Lysobacter species. The six L. capsici strains tested developed dendrite-like colonies when grown on jellified pea broth, while the development of dendrite-like colonies was not recorded in the media commonly used in motility assays. To determine the presence of genes responsible for biogenesis of the flagellum and type IV pili, the genome of L. capsici AZ78 was mined. Genes encoding structural components an d regulatory factors of type IV pili were upregulated in L. capsici AZ78 cells grown on the above-mentioned medium, as compared with the other tested media. These results provide new insight into the motility mechanism of L. capsici members and the role of type IV pili and pea compounds on the epiphytic fitness and biocontrol features of L. capsici AZ78.

  17. An immunoperoxidase study of laminin and type IV collagen distribution in carcinoma of the cervix and vulva.

    Science.gov (United States)

    Ehrmann, R L; Dwyer, I M; Yavner, D; Hancock, W W

    1988-08-01

    Basement membrane immunostaining was performed on pepsin-digested, paraffin-embedded blocks of 29 squamous cell carcinomas of the cervix (invasive and in situ) and 13 of the vulva, using polyclonal rabbit antibodies to human laminin and type IV collagen, both staining identically. Laminin with varying defectiveness surrounded invasive foci, whereas adjacent carcinoma in situ or normal epithelium had intact laminin. The amount of laminin usually reflected the degree of tumor differentiation. Absence of laminin around totally keratinized or necrotic tumor nests indicated its dependency on viable cells. New buds from established invasive tumor nests were often more laminin-defective than the parent nest and suggested a cyclic invasive process, with laminin loss during a growth surge followed by laminin reformation during quiescence. In cases of questionable early stromal invasion, deficient laminin could sway the decision toward making a positive diagnosis. The tendency of laminin gaps and tumor buds to contain large malignant cells with pleomorphic nuclei supports the concept of a change in tumor cell metabolism during active invasion. Laminin also appeared around metastatic tumor within lymph nodes. The relationship of inflammation to tumor laminin defectiveness varied.

  18. Molecular and Genetic Analysis of Collagen Type IV Mutant Mouse Models of Spontaneous Intracerebral Hemorrhage Identify Mechanisms for Stroke Prevention

    Science.gov (United States)

    Jeanne, Marion; Jorgensen, Jeff; Gould, Douglas B.

    2015-01-01

    Background Collagen type IV alpha 1 (COL4A1) and alpha 2 (COL4A2) form heterotrimers critical for vascular basement membrane stability and function. Patients with COL4A1 or COL4A2 mutations suffer from diverse cerebrovascular diseases including cerebral microbleeds, porencephaly and fatal intracerebral hemorrhage (ICH). However, the pathogenic mechanisms remain unknown and there is a lack of effective treatment. Methods and Results Using Col4a1 and Col4a2 mutant mouse models, we investigated the genetic complexity and cellular mechanisms underlying the disease. We found that Col4a1 mutations cause abnormal vascular development, which triggers small vessel disease, recurrent hemorrhagic strokes and age-related macro-angiopathy. We showed that allelic heterogeneity, genetic context and environmental factors, such as acute exercise or anticoagulant medication, modulated disease severity and contributed to phenotypic heterogeneity. We found that intracellular accumulation of mutant collagen in vascular endothelial cells and pericytes was a key triggering factor of ICH. Finally, we showed that treatment of mutant mice with a FDA-approved chemical chaperone resulted in a decreased collagen intracellular accumulation and a significant reduction of ICH severity. Conclusions Our data are the first to show therapeutic prevention in vivo of ICH due to Col4a1 mutation, and imply that a mechanism-based therapy promoting protein folding might also prevent ICH in patients with COL4A1 and COL4A2 mutations. PMID:25753534

  19. Vitamin D deficiency and rickets in children and adolescents with ichthyosiform erythroderma in type IV and V skin.

    Science.gov (United States)

    Chouhan, K; Sethuraman, G; Gupta, N; Sharma, V K; Kabra, M; Khaitan, B K; Sreenivas, V; Ramam, M; Kusumakar, S; Thulkar, S; Paller, A S

    2012-03-01

    Ichthyosiform erythroderma due to keratinizing disorders may suppress cutaneous vitamin D synthesis, leading to vitamin D deficiency and rickets. To determine the prevalence of vitamin D deficiency and rickets in children and adolescents with congenital ichthyosis and other keratinizing disorders with erythroderma and scaling. In this cross-sectional study, 45 children and adolescents with ichthyosiform erythroderma due to keratinizing disorders, and 66 controls (group 1: age and sex matched, with skin diseases other than keratinizing disorders; group 2: age and sex matched, healthy volunteers) were included. Evidence of rickets was determined clinically (physical examination and radiographs) and biochemically {serum calcium, phosphorus, alkaline phosphatase, 25-hydroxy vitamin D [25(OH)D] and parathyroid hormone (PTH)}. All patients in the disease group had clinical, radiological or biochemical evidence of rickets [25(OH)D65pgmL(-1) ) was also significantly higher in the disease group than in controls (PChildren and adolescents with various forms of ichthyosiform erythroderma, especially those with pigmented skin (types IV-VI), are at increased risk of developing vitamin D deficiency and clinical rickets. © 2011 The Authors. BJD © 2011 British Association of Dermatologists.

  20. The BID Domain of Type IV Secretion Substrates Forms a Conserved Four-Helix Bundle Topped with a Hook.

    Science.gov (United States)

    Stanger, Frédéric V; de Beer, Tjaart A P; Dranow, David M; Schirmer, Tilman; Phan, Isabelle; Dehio, Christoph

    2017-01-03

    The BID (Bep intracellular delivery) domain functions as secretion signal in a subfamily of protein substrates of bacterial type IV secretion (T4S) systems. It mediates transfer of (1) relaxases and the attached DNA during bacterial conjugation, and (2) numerous Bartonella effector proteins (Beps) during protein transfer into host cells infected by pathogenic Bartonella species. Furthermore, BID domains of Beps have often evolved secondary effector functions within host cells. Here, we provide crystal structures for three representative BID domains and describe a novel conserved fold characterized by a compact, antiparallel four-helix bundle topped with a hook. The conserved hydrophobic core provides a rigid scaffold to a surface that, despite a few conserved exposed residues and similarities in charge distribution, displays significant variability. We propose that the genuine function of BID domains as T4S signal may primarily depend on their rigid structure, while the plasticity of their surface may facilitate adaptation to secondary effector functions. Copyright © 2016 Elsevier Ltd. All rights reserved.

  1. Key components of the eight classes of type IV secretion systems involved in bacterial conjugation or protein secretion.

    Science.gov (United States)

    Guglielmini, Julien; Néron, Bertrand; Abby, Sophie S; Garcillán-Barcia, María Pilar; de la Cruz, Fernando; Rocha, Eduardo P C

    2014-05-01

    Conjugation of DNA through a type IV secretion system (T4SS) drives horizontal gene transfer. Yet little is known on the diversity of these nanomachines. We previously found that T4SS can be divided in eight classes based on the phylogeny of the only ubiquitous protein of T4SS (VirB4). Here, we use an ab initio approach to identify protein families systematically and specifically associated with VirB4 in each class. We built profiles for these proteins and used them to scan 2262 genomes for the presence of T4SS. Our analysis led to the identification of thousands of occurrences of 116 protein families for a total of 1623 T4SS. Importantly, we could identify almost always in our profiles the essential genes of well-studied T4SS. This allowed us to build a database with the largest number of T4SS described to date. Using profile-profile alignments, we reveal many new cases of homology between components of distant classes of T4SS. We mapped these similarities on the T4SS phylogenetic tree and thus obtained the patterns of acquisition and loss of these protein families in the history of T4SS. The identification of the key VirB4-associated proteins paves the way toward experimental analysis of poorly characterized T4SS classes. © The Author(s) 2014. Published by Oxford University Press.

  2. Common requirement for the relaxosome of plasmid R1 in multiple activities of the conjugative type IV secretion system.

    Science.gov (United States)

    Lang, Silvia; Gruber, Christian J; Raffl, Sandra; Reisner, Andreas; Zechner, Ellen L

    2014-06-01

    Macromolecular transport by bacterial type IV secretion systems involves regulated uptake of (nucleo)protein complexes by the cell envelope-spanning transport channel. A coupling protein receptor is believed to recognize the specific proteins destined for transfer, but the steps initiating their translocation remain unknown. Here, we investigate the contribution of a complex of transfer initiation proteins, the relaxosome, of plasmid R1 to translocation of competing transferable substrates from mobilizable plasmids ColE1 and CloDF13 or the bacteriophage R17. We found that not only does the R1 translocation machinery engage the R1 relaxosome during conjugative self-transfer and during infection by R17 phage but it is also activated by its cognate relaxosome to mediate the export of an alternative plasmid. Transporter activity was optimized by the R1 relaxosome even when this complex itself could not be transferred, i.e., when the N-terminal activation domain (amino acids 1 to 992 [N1-992]) of TraI was present without the C-terminal conjugative helicase domain. We propose that the functional dependence of the transfer machinery on the R1 relaxosome for initiating translocation ensures that dissemination of heterologous plasmids does not occur at the expense of self-transfer.

  3. Twitching motility of bacteria with type-IV pili: Fractal walks, first passage time, and their consequences on microcolonies

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    Bisht, Konark; Klumpp, Stefan; Banerjee, Varsha; Marathe, Rahul

    2017-11-01

    A human pathogen, Neisseria gonorrhoeae (NG), moves on surfaces by attaching and retracting polymeric structures called Type IV pili. The tug-of-war between the pili results in a two-dimensional stochastic motion called twitching motility. In this paper, with the help of real-time NG trajectories, we develop coarse-grained models for their description. The fractal properties of these trajectories are determined and their influence on first passage time and formation of bacterial microcolonies is studied. Our main observations are as follows: (i) NG performs a fast ballistic walk on small time scales and a slow diffusive walk over long time scales with a long crossover region; (ii) there exists a characteristic persistent length lp*, which yields the fastest growth of bacterial aggregates or biofilms. Our simulations reveal that lp*˜L0.6 , where L ×L is the surface on which the bacteria move; (iii) the morphologies have distinct fractal characteristics as a consequence of the ballistic and diffusive motion of the constituting bacteria.

  4. Cleavage of phosphorothioated DNA and methylated DNA by the type IV restriction endonuclease ScoMcrA.

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    Guang Liu

    2010-12-01

    Full Text Available Many taxonomically diverse prokaryotes enzymatically modify their DNA by replacing a non-bridging oxygen with a sulfur atom at specific sequences. The biological implications of this DNA S-modification (phosphorothioation were unknown. We observed that simultaneous expression of the dndA-E gene cluster from Streptomyces lividans 66, which is responsible for the DNA S-modification, and the putative Streptomyces coelicolor A(32 Type IV methyl-dependent restriction endonuclease ScoA3McrA (Sco4631 leads to cell death in the same host. A His-tagged derivative of ScoA3McrA cleaved S-modified DNA and also Dcm-methylated DNA in vitro near the respective modification sites. Double-strand cleavage occurred 16-28 nucleotides away from the phosphorothioate links. DNase I footprinting demonstrated binding of ScoA3McrA to the Dcm methylation site, but no clear binding could be detected at the S-modified site under cleavage conditions. This is the first report of in vitro endonuclease activity of a McrA homologue and also the first demonstration of an enzyme that specifically cleaves S-modified DNA.

  5. Aging is associated with increased collagen type IV accumulation in the basal lamina of human cerebral microvessels

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    Danek Adrian

    2004-09-01

    Full Text Available Abstract Background Microvascular alterations contribute to the development of stroke and vascular dementia. The goal of this study was to evaluate age and hypertension related changes of the basal lamina in cerebral microvessels of individuals, who died from non-cerebral causes. Results We examined 27 human brains: 11 young and 16 old patients. Old patients were divided into two subgroups, those with hypertension (n = 8 and those without hypertension (n = 8. Basal lamina changes of the cerebral microvessels were determined in the putamen using antibodies against collagen type IV and by quantitative analysis of vessel number, total stained area of collagen, thickness of the vessel wall and lumen, and relative staining intensity using immunofluorescence. The total number of collagen positive vessels per microscopic field was reduced in old compared to young subjects (12.0+/-0.6 vs. 15.1+/-1.2, p = 0.02. The relative collagen content per vessel (1.01+/-0.06 vs. 0.76+/-0.05, p = 0.01 and the relative collagen intensity (233.1+/-4.5 vs. 167.8+/-10.6, p Conclusions The present data show age-related changes of the cerebral microvessels in sections of human putamen for the first time. Due to the accumulation of collagen, microvessels thicken and show a reduction in their lumen. Besides this, the number of vessels decreases. These findings might represent a precondition for the development of vascular cognitive impairment. However, hypertension was not proven to modulate these changes.

  6. Aging is associated with increased collagen type IV accumulation in the basal lamina of human cerebral microvessels.

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    Uspenskaia, Olga; Liebetrau, Martin; Herms, Jochen; Danek, Adrian; Hamann, Gerhard F

    2004-09-24

    Microvascular alterations contribute to the development of stroke and vascular dementia. The goal of this study was to evaluate age and hypertension related changes of the basal lamina in cerebral microvessels of individuals, who died from non-cerebral causes. We examined 27 human brains: 11 young and 16 old patients. Old patients were divided into two subgroups, those with hypertension (n = 8) and those without hypertension (n = 8). Basal lamina changes of the cerebral microvessels were determined in the putamen using antibodies against collagen type IV and by quantitative analysis of vessel number, total stained area of collagen, thickness of the vessel wall and lumen, and relative staining intensity using immunofluorescence. The total number of collagen positive vessels per microscopic field was reduced in old compared to young subjects (12.0+/-0.6 vs. 15.1+/-1.2, p = 0.02). The relative collagen content per vessel (1.01+/-0.06 vs. 0.76+/-0.05, p = 0.01) and the relative collagen intensity (233.1+/-4.5 vs. 167.8+/-10.6, p hypertensive and non-hypertensive patients. The present data show age-related changes of the cerebral microvessels in sections of human putamen for the first time. Due to the accumulation of collagen, microvessels thicken and show a reduction in their lumen. Besides this, the number of vessels decreases. These findings might represent a precondition for the development of vascular cognitive impairment. However, hypertension was not proven to modulate these changes.

  7. Computational prediction of secretion systems and secretomes of Brucella: identification of novel type IV effectors and their interaction with the host.

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    Sankarasubramanian, Jagadesan; Vishnu, Udayakumar S; Dinakaran, Vasudevan; Sridhar, Jayavel; Gunasekaran, Paramasamy; Rajendhran, Jeyaprakash

    2016-01-01

    Brucella spp. are facultative intracellular pathogens that cause brucellosis in various mammals including humans. Brucella survive inside the host cells by forming vacuoles and subverting host defence systems. This study was aimed to predict the secretion systems and the secretomes of Brucella spp. from 39 complete genome sequences available in the databases. Furthermore, an attempt was made to identify the type IV secretion effectors and their interactions with host proteins. We predicted the secretion systems of Brucella by the KEGG pathway and SecReT4. Brucella secretomes and type IV effectors (T4SEs) were predicted through genome-wide screening using JVirGel and S4TE, respectively. Protein-protein interactions of Brucella T4SEs with their hosts were analyzed by HPIDB 2.0. Genes coding for Sec and Tat pathways of secretion and type I (T1SS), type IV (T4SS) and type V (T5SS) secretion systems were identified and they are conserved in all the species of Brucella. In addition to the well-known VirB operon coding for the type IV secretion system (T4SS), we have identified the presence of additional genes showing homology with T4SS of other organisms. On the whole, 10.26 to 14.94% of total proteomes were found to be either secreted (secretome) or membrane associated (membrane proteome). Approximately, 1.7 to 3.0% of total proteomes were identified as type IV secretion effectors (T4SEs). Prediction of protein-protein interactions showed 29 and 36 host-pathogen specific interactions between Bos taurus (cattle)-B. abortus and Ovis aries (sheep)-B. melitensis, respectively. Functional characterization of the predicted T4SEs and their interactions with their respective hosts may reveal the secrets of host specificity of Brucella.

  8. Parallel evolution of a type IV secretion system in radiating lineages of the host-restricted bacterial pathogen Bartonella.

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    Engel, Philipp; Salzburger, Walter; Liesch, Marius; Chang, Chao-Chin; Maruyama, Soichi; Lanz, Christa; Calteau, Alexandra; Lajus, Aurélie; Médigue, Claudine; Schuster, Stephan C; Dehio, Christoph

    2011-02-10

    Adaptive radiation is the rapid origination of multiple species from a single ancestor as the result of concurrent adaptation to disparate environments. This fundamental evolutionary process is considered to be responsible for the genesis of a great portion of the diversity of life. Bacteria have evolved enormous biological diversity by exploiting an exceptional range of environments, yet diversification of bacteria via adaptive radiation has been documented in a few cases only and the underlying molecular mechanisms are largely unknown. Here we show a compelling example of adaptive radiation in pathogenic bacteria and reveal their genetic basis. Our evolutionary genomic analyses of the α-proteobacterial genus Bartonella uncover two parallel adaptive radiations within these host-restricted mammalian pathogens. We identify a horizontally-acquired protein secretion system, which has evolved to target specific bacterial effector proteins into host cells as the evolutionary key innovation triggering these parallel adaptive radiations. We show that the functional versatility and adaptive potential of the VirB type IV secretion system (T4SS), and thereby translocated Bartonella effector proteins (Beps), evolved in parallel in the two lineages prior to their radiations. Independent chromosomal fixation of the virB operon and consecutive rounds of lineage-specific bep gene duplications followed by their functional diversification characterize these parallel evolutionary trajectories. Whereas most Beps maintained their ancestral domain constitution, strikingly, a novel type of effector protein emerged convergently in both lineages. This resulted in similar arrays of host cell-targeted effector proteins in the two lineages of Bartonella as the basis of their independent radiation. The parallel molecular evolution of the VirB/Bep system displays a striking example of a key innovation involved in independent adaptive processes and the emergence of bacterial pathogens

  9. Parallel evolution of a type IV secretion system in radiating lineages of the host-restricted bacterial pathogen Bartonella.

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    Philipp Engel

    2011-02-01

    Full Text Available Adaptive radiation is the rapid origination of multiple species from a single ancestor as the result of concurrent adaptation to disparate environments. This fundamental evolutionary process is considered to be responsible for the genesis of a great portion of the diversity of life. Bacteria have evolved enormous biological diversity by exploiting an exceptional range of environments, yet diversification of bacteria via adaptive radiation has been documented in a few cases only and the underlying molecular mechanisms are largely unknown. Here we show a compelling example of adaptive radiation in pathogenic bacteria and reveal their genetic basis. Our evolutionary genomic analyses of the α-proteobacterial genus Bartonella uncover two parallel adaptive radiations within these host-restricted mammalian pathogens. We identify a horizontally-acquired protein secretion system, which has evolved to target specific bacterial effector proteins into host cells as the evolutionary key innovation triggering these parallel adaptive radiations. We show that the functional versatility and adaptive potential of the VirB type IV secretion system (T4SS, and thereby translocated Bartonella effector proteins (Beps, evolved in parallel in the two lineages prior to their radiations. Independent chromosomal fixation of the virB operon and consecutive rounds of lineage-specific bep gene duplications followed by their functional diversification characterize these parallel evolutionary trajectories. Whereas most Beps maintained their ancestral domain constitution, strikingly, a novel type of effector protein emerged convergently in both lineages. This resulted in similar arrays of host cell-targeted effector proteins in the two lineages of Bartonella as the basis of their independent radiation. The parallel molecular evolution of the VirB/Bep system displays a striking example of a key innovation involved in independent adaptive processes and the emergence of bacterial

  10. Discovery of JANUVIA (Sitagliptin), a selective dipeptidyl peptidase IV inhibitor for the treatment of type 2 diabetes.

    Science.gov (United States)

    Thornberry, Nancy A; Weber, Ann E

    2007-01-01

    The emergence of glucagon-like peptide 1 (GLP-1) as a well validated approach to the treatment of type 2 diabetes and preclinical validation of dipeptidyl peptidase IV (DPP-4) inhibition as an alternate, oral approach to GLP-1 therapy prompted the initiation of a DPP-4 inhibitor program at Merck in 1999. DPP-4 inhibitors threo- and allo-isoleucyl thiazolidide were in-licensed to jump start the program; however, development was discontinued due to profound toxicity in rat and dog safety studies. The observation that both compounds inhibit the related proline peptidases DPP8 and DPP9 led to the hypothesis that inhibition of DPP8 and/or DPP9 could evoke severe toxicities in preclinical species. Indeed, the observed toxicities were recapitulated with a selective dual DPP8/9 inhibitor but not with an inhibitor selective for DPP-4. Thus, medicinal chemistry efforts focused on identifying a highly selective DPP-4 inibitor for clinical development. Initial work in an alpha-amino acid series related to isoleucyl thiazolidide was discontinued due to lack of selectivity; however, SAR studies on two screening leads led to the identification of a highly selective beta-amino acid piperazine series. In an effort to stabilize the piperazine moiety, which was extensively metabolized in vivo, a series of bicyclic derivatives were prepared, culminating in the identification of a potent and selective triazolopiperazine series. Unlike their monocyclic counterparts, these analogs typically showed excellent pharmacokinetic properties in preclinical species. Optimization of this series led to the discovery of JANUVIA (sitagliptin), a highly selective DPP-4 inhibitor for the treatment of type 2 diabetes.

  11. SCCmec-associated psm-mec mRNA promotes Staphylococcus epidermidis biofilm formation.

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    Yang, Yongchang; Zhang, Xuemei; Huang, Wenfang; Yin, Yibing

    2016-10-01

    Biofilm formation is considered the major pathogenic mechanism of Staphylococcus epidermidis-associated nosocomial infections. Reports have shown that SCCmec-associated psm-mec regulated methicillin-resistant Staphylococcus aureus virulence and biofilm formation. However, the role of psm-mec in S. epidermidis remains unclear. To this purpose, we analysed 165 clinical isolates of S. epidermidis to study the distribution, mutation and expression of psm-mec and the relationship between this gene and biofilm formation. Next, we constructed three psm-mec deletion mutants, one psm-mec transgene expression strain (p221) and two psm-mec point mutant strains (pM, pAG) to explore its effects on S. epidermidis biofilm formation. Then, the amount of biofilm formation, extracellular DNA (eDNA) and Triton X-100-induced autolysis of the constructed strains was measured. Results of psm-mec deletion and transgene expression showed that the gene regulated S. epidermidis biofilm formation. Compared with the control strains, the ability to form biofilm, Triton X-100-induced autolysis and the amount of eDNA increased in the p221 strain and the two psm-mec mutants pM and pAG expressed psm-mec mRNA without its protein, whereas no differences were observed among the three constructed strains, illustrating that psm-mec mRNA promoted S. epidermidis biofilm formation through up-regulation of bacterial autolysis and the release of eDNA. Our results reveal that acquisition of psm-mec promotes S. epidermidis biofilm formation.

  12. ANTI-COLLAGEN TYPE IV ANTIBODIES AND THE DEVELOPMENT OF MICROVASCULAR COMPLICATIONS IN DIABETIC PATIENTS WITH ARTERIAL HYPERTENSION

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    Asparuh G. Nikolov

    2012-11-01

    Full Text Available Background and Aims: Thickening of basement membrane in capillaries and small vessels is a well-known finding and important in the progression of diabetic microangiopathy. Patients with diabetes mellitus and arterial hypertension are at higher risk of vascular disease. Material and methods: To monitor the metabolism of the basement membrane protein collagen type IV (CIV in type 2 diabetes mellitus (T2DM, serum levels of antibodies to CIV (ACIV IgG, IgM and IgA were measured using an ELISA method in 93 patients with type 2 diabetes mellitus and arterial hypertension (AH (mean age 61,4±11,3 years, diabetes duration 9,88±3,12 years; hypertension duration 9,28±4,98. These values were compared to serum antibodies to CIV in 42 age and sex matched controls. Diabetics were divided in two groups according to presence- Group 1 (n=67 or absence- Group 2 (n=26 of microangiopathy. Results: Patients with T2DM and AH showed statistically significant higher levels of ACIV IgG in comparison to healthy controls (0.30±0.12 vs. 0.21±0.08 (p=0.0001. Group 1 showed significantly hihger levels of ACIV IgG than Group 2 (0.32±0.13vs. 0.24±0.08 (p=0.009 and healthy controls (0.32±0.13vs. 0.21±0.08 (p=0.0001. ACIV IgG are statistically significant higher in diabetics with retinopathy than this without (0.33±0.10 vs. 0.26±0.13 (р=0.04. ACIV IgG correlates with diabetes duration (r=0.49; (p=0.0004, retinopathy (r=0.20; (p=0.05 and BMI (r=-0.24; (p=0.05. Serum ACIV IgM and IgA levels in patients with T2DM and AH were lower than these in controls, but the differences are not statistically significant.Conclusion: Our study showed a relationship between elevation of serum levels of ACIV IgG in diabetics and development of microangiopathy.

  13. Phylogenomics and sequence-structure-function relationships in the GmrSD family of Type IV restriction enzymes.

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    Machnicka, Magdalena A; Kaminska, Katarzyna H; Dunin-Horkawicz, Stanislaw; Bujnicki, Janusz M

    2015-10-23

    GmrSD is a modification-dependent restriction endonuclease that specifically targets and cleaves glucosylated hydroxymethylcytosine (glc-HMC) modified DNA. It is encoded either as two separate single-domain GmrS and GmrD proteins or as a single protein carrying both domains. Previous studies suggested that GmrS acts as endonuclease and NTPase whereas GmrD binds DNA. In this work we applied homology detection, sequence conservation analysis, fold recognition and homology modeling methods to study sequence-structure-function relationships in the GmrSD restriction endonucleases family. We also analyzed the phylogeny and genomic context of the family members. Results of our comparative genomics study show that GmrS exhibits similarity to proteins from the ParB/Srx fold which can have both NTPase and nuclease activity. In contrast to the previous studies though, we attribute the nuclease activity also to GmrD as we found it to contain the HNH endonuclease motif. We revealed residues potentially important for structure and function in both domains. Moreover, we found that GmrSD systems exist predominantly as a fused, double-domain form rather than as a heterodimer and that their homologs are often encoded in regions enriched in defense and gene mobility-related elements. Finally, phylogenetic reconstructions of GmrS and GmrD domains revealed that they coevolved and only few GmrSD systems appear to be assembled from distantly related GmrS and GmrD components. Our study provides insight into sequence-structure-function relationships in the yet poorly characterized family of Type IV restriction enzymes. Comparative genomics allowed to propose possible role of GmrD domain in the function of the GmrSD enzyme and possible active sites of both GmrS and GmrD domains. Presented results can guide further experimental characterization of these enzymes.

  14. Camel milk attenuates the biochemical and morphological features of diabetic nephropathy: inhibition of Smad1 and collagen type IV synthesis.

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    Korish, Aida A; Abdel Gader, Abdel Galil; Korashy, Hesham M; Al-Drees, Abdul Majeed; Alhaider, Abdulqader A; Arafah, Maha M

    2015-03-05

    Diabetic nephropathy (DN) is a common microvascular complication of diabetes mellitus (DM) that worsens its morbidity and mortality. There is evidence that camel milk (CM) improves the glycemic control in DM but its effect on the renal complications especially the DN remains unclear. Thus the current study aimed to characterize the effects of CM treatment on streptozotocin (STZ)-induced DN. Using STZ-induced diabetes, we investigated the effect of CM treatment on kidney function, proteinuria, renal Smad1, collagen type IV (Col4), blood glucose, insulin resistance (IR), lipid peroxidation, the antioxidant superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH). In addition renal morphology was also examined. The current results showed that rats with untreated diabetes exhibited marked hyperglycemia, IR, high serum urea and creatinine levels, excessive proteinuria, increased renal Smad1 and Col4, glomerular expansion, and extracellular matrix deposition. There was also increased lipid peroxidation products, decreased antioxidant enzyme activity and GSH levels. Camel milk treatment decreased blood glucose, IR, and lipid peroxidation. Superoxide dismutase and CAT expression, CAT activity, and GSH levels were increased. The renoprotective effects of CM were demonstrated by the decreased serum urea and creatinine, proteinuria, Smad1, Col4, and preserved normal tubulo-glomerular morphology. In conclusion, beside its hypoglycemic action, CM attenuates the early changes of DN, decreased renal Smad1 and Col4. This could be attributed to a primary action on the glomerular mesangial cells, or secondarily to the hypoglycemic and antioxidant effects of CM. The protective effects of CM against DN support its use as an adjuvant anti-diabetes therapy. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  15. Peptide Nucleic Acid Knockdown and Intra-host Cell Complementation of Ehrlichia Type IV Secretion System Effector.

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    Sharma, Pratibha; Teymournejad, Omid; Rikihisa, Yasuko

    2017-01-01

    Survival of Ehrlichia chaffeensis depends on obligatory intracellular infection. One of the barriers to E. chaffeensis research progress has been the inability, using conventional techniques, to generate knock-out mutants for genes essential for intracellular infection. This study examined the use of Peptide Nucleic Acids (PNAs) technology to interrupt type IV secretion system (T4SS) effector protein expression in E. chaffeensis followed by intracellular complementation of the effector to determine its requirement for infection. Successful E. chaffeensis infection depends on the E. chaffeensis-specific T4SS protein effector, ehrlichial translocated factor-1 (Etf-1), which induces Rab5-regulated autophagy to provide host cytosolic nutrients required for E. chaffeensis proliferation. Etf-1 is also imported by host cell mitochondria where it inhibits host cell apoptosis to prolong its infection. We designed a PNA specific to Etf-1 and showed that the PNA bound to the target region of single-stranded Etf-1 RNA using a competitive binding assay. Electroporation of E. chaffeensis with this PNA significantly reduced Etf-1 mRNA and protein, and the bacteria's ability to induce host cell autophagy and infect host cells. Etf-1 PNA-mediated inhibition of ehrlichial Etf-1 expression and E. chaffeensis infection could be intracellularly trans-complemented by ectopic expression of Etf-1-GFP in host cells. These data affirmed the critical role of bacterial T4SS effector in host cell autophagy and E. chaffeensis infection, and demonstrated the use of PNA to analyze the gene functions of obligate intracellular bacteria.

  16. Identification of ElpA, a Coxiella burnetii pathotype-specific Dot/Icm type IV secretion system substrate.

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    Graham, Joseph G; Winchell, Caylin G; Sharma, Uma M; Voth, Daniel E

    2015-03-01

    Coxiella burnetii causes human Q fever, a zoonotic disease that presents with acute flu-like symptoms and can result in chronic life-threatening endocarditis. In human alveolar macrophages, C. burnetii uses a Dot/Icm type IV secretion system (T4SS) to generate a phagolysosome-like parasitophorous vacuole (PV) in which to replicate. The T4SS translocates effector proteins, or substrates, into the host cytosol, where they mediate critical cellular events, including interaction with autophagosomes, PV formation, and prevention of apoptosis. Over 100 C. burnetii Dot/Icm substrates have been identified, but the function of most remains undefined. Here, we identified a novel Dot/Icm substrate-encoding open reading frame (CbuD1884) present in all C. burnetii isolates except the Nine Mile reference isolate, where the gene is disrupted by a frameshift mutation, resulting in a pseudogene. The CbuD1884 protein contains two transmembrane helices (TMHs) and a coiled-coil domain predicted to mediate protein-protein interactions. The C-terminal region of the protein contains a predicted Dot/Icm translocation signal and was secreted by the T4SS, while the N-terminal portion of the protein was not secreted. When ectopically expressed in eukaryotic cells, the TMH-containing N-terminal region of the CbuD1884 protein trafficked to the endoplasmic reticulum (ER), with the C terminus dispersed nonspecifically in the host cytoplasm. This new Dot/Icm substrate is now termed ElpA (ER-localizing protein A). Full-length ElpA triggered substantial disruption of ER structure and host cell secretory transport. These results suggest that ElpA is a pathotype-specific T4SS effector that influences ER function during C. burnetii infection. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  17. The Legionella pneumophila IcmSW complex interacts with multiple Dot/Icm effectors to facilitate type IV translocation.

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    Eric D Cambronne

    2007-12-01

    Full Text Available Many gram-negative pathogens use a type IV secretion system (T4SS to deliver effector proteins into eukaryotic host cells. The fidelity of protein translocation depends on the efficient recognition of effector proteins by the T4SS. Legionella pneumophila delivers a large number of effector proteins into eukaryotic cells using the Dot/Icm T4SS. How the Dot/Icm system is able to recognize and control the delivery of effectors is poorly understood. Recent studies suggest that the IcmS and IcmW proteins interact to form a stable complex that facilitates translocation of effector proteins by the Dot/Icm system by an unknown mechanism. Here we demonstrate that the IcmSW complex is necessary for the productive translocation of multiple Dot/Icm effector proteins. Effector proteins that were able to bind IcmSW in vitro required icmS and icmW for efficient translocation into eukaryotic cells during L. pneumophila infection. We identified regions in the effector protein SidG involved in icmSW-dependent translocation. Although the full-length SidG protein was translocated by an icmSW-dependent mechanism, deletion of amino terminal regions in the SidG protein resulted in icmSW-independent translocation, indicating that the IcmSW complex is not contributing directly to recognition of effector proteins by the Dot/Icm system. Biochemical and genetic studies showed that the IcmSW complex interacts with a central region of the SidG protein. The IcmSW interaction resulted in a conformational change in the SidG protein as determined by differences in protease sensitivity in vitro. These data suggest that IcmSW binding to effectors could enhance effector protein delivery by mediating a conformational change that facilitates T4SS recognition of a translocation domain located in the carboxyl region of the effector protein.

  18. Evidence of transfer by conjugation of type IV secretion system genes between Bartonella species and Rhizobium radiobacter in amoeba.

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    Watcharee Saisongkorh

    Full Text Available BACKGROUND: Bartonella species cospeciate with mammals and live within erythrocytes. Even in these specific niches, it has been recently suggested by bioinformatic analysis of full genome sequences that Lateral Gene Transfer (LGT may occur but this has never been demonstrated biologically. Here we describe the sequence of the B. rattaustraliani (AUST/NH4(T circular plasmid (pNH4 that encodes the tra cluster of the Type IV secretion system (T4SS and we eventually provide evidence that Bartonella species may conjugate and exchange this plasmid inside amoeba. PRINCIPAL FINDINGS: The T4SS of pNH4 is critical for intracellular viability of bacterial pathogens, exhibits bioinformatic evidence of LGT among bacteria living in phagocytic protists. For instance, 3 out of 4 T4SS encoding genes from pNH4 appear to be closely related to Rhizobiales, suggesting that gene exchange occurs between intracellular bacteria from mammals (bartonellae and plants (Rhizobiales. We show that B. rattaustraliani and Rhizobium radiobacter both survived within the amoeba Acanthamoeba polyphaga and can conjugate together. Our findings further support the hypothesis that tra genes might also move into and out of bacterial communities by conjugation, which might be the primary means of genomic evolution for intracellular adaptation by cross-talk of interchangeable genes between Bartonella species and plant pathogens. CONCLUSIONS: Based on this, we speculate that amoeba favor the transfer of genes as phagocytic protists, which allows for intraphagocytic survival and, as a consequence, promotes the creation of potential pathogenic organisms.

  19. Characterization of type IV antifreeze gene in Nile tilapia (Oreochromis niloticus) and influence of cold and hot weather on its expression and some immune-related genes.

    Science.gov (United States)

    Ammar, Asmma Y; El Nahas, Abeer F; Mahmoud, Shawky; Barakat, Mohamed E; Hassan, Asmaa M

    2017-12-12

    The aim of this work is to study the effect of the thermal stress of ambient temperature during winter and summer on the expression of type IV antifreeze gene (ANF IV) in different tissues of Nile tilapia (Oreochromis niloticus) as well as some immune-related genes. At first, genomic ANF IV gene was characterized from one fish; 124 amino acids were identified with 92.7% similarity with that on the gene bank. Expression of ANF IV and immune-related genes were done twice, once at the end of December (winter sample, temperature 14 °C) and the other at August (summer sample, temperature 36 °C). Assessment of ANF IV gene expression in different organs of fish was done; splenic mRNA was used for assessment of immune-related gene transcripts (CXCl2 chemokine, cc-chemokine, INF-3A, and MHC IIβ). Winter expression analysis of AFP IV in O. niloticus revealed significant upregulation of mRNA transcript levels in the intestine, gills, skin, spleen, liver, and brain with 324.03-, 170.06-, 107.63-, 97.61-, 94.35-, and 27.85-folds, respectively. Furthermore, upregulation in the gene was observed in some organs during summer: in the liver, gills, skin, intestine, and brain with lower levels compared with winter. The level of expression of immune-related genes in winter is significantly higher than summer in all assessed genes. Cc-chemokine gene expression was the most affected in both winter and summer. Variable expression profile of ANF IV in different organs and in different seasons together with its amino acid similarity of N-terminal and C-terminal with apolipoprotein (lipid binder) and form of high-density lipoprotein (HDL) suggests a different role for this protein which may be related to lipid metabolism.

  20. L-arginine mediated renaturation enhances yield of human, α6 Type IV collagen non-collagenous domain from bacterial inclusion bodies.

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    Gunda, Venugopal; Boosani, Chandra Shekhar; Verma, Raj Kumar; Guda, Chittibabu; Sudhakar, Yakkanti Akul

    2012-10-01

    The anti-angiogenic, carboxy terminal non-collagenous domain (NC1) derived from human Collagen type IV alpha 6 chain, [α6(IV)NC1] or hexastatin, was earlier obtained using different recombinant methods of expression in bacterial systems. However, the effect of L-arginine mediated renaturation in enhancing the relative yields of this protein from bacterial inclusion bodies has not been evaluated. In the present study, direct stirring and on-column renaturation methods using L-arginine and different size exclusion chromatography matrices were applied for enhancing the solubility in purifying the recombinant α6(IV)NC1 from bacterial inclusion bodies. This methodology enabled purification of higher quantities of soluble protein from inclusion bodies, which inhibited endothelial cell proliferation, migration and tube formation. Thus, the scope for L-arginine mediated renaturation in obtaining higher yields of soluble, biologically active NC1 domain from bacterial inclusion bodies was evaluated.

  1. Spectroscopic Studies of Abiotic and Biological Nanomaterials: Silver Nanoparticles, Rhodamine 6G Adsorbed on Graphene, and c-Type Cytochromes and Type IV Pili in Geobacter sulfurreducens

    Science.gov (United States)

    Thrall, Elizabeth S.

    This thesis describes spectroscopic studies of three different systems: silver nanoparticles, the dye molecule rhodamine 6G adsorbed on graphene, and the type IV pili and c-type cytochromes produced by the dissimilatory metal-reducing bacterium Geobacter sulfurreducens. Although these systems are quite different in some ways, they can all be considered examples of nanomaterials. A nanomaterial is generally defined as having at least one dimension below 100 nm in size. Silver nanoparticles, with sub-100 nm size in all dimensions, are examples of zero-dimensional nanomaterials. Graphene, a single atomic layer of carbon atoms, is the paradigmatic two-dimensional nanomaterial. And although bacterial cells are on the order of 1 μm in size, the type IV pili and multiheme c-type cytochromes produced by G. sulfurreducens can be considered to be one- and zero-dimensional nanomaterials respectively. A further connection between these systems is their strong interaction with visible light, allowing us to study them using similar spectroscopic tools. The first chapter of this thesis describes research on the plasmon-mediated photochemistry of silver nanoparticles. Silver nanoparticles support coherent electron oscillations, known as localized surface plasmons, at resonance frequencies that depend on the particle size and shape and the local dielectric environment. Nanoparticle absorption and scattering cross-sections are maximized at surface plasmon resonance frequencies, and the electromagnetic field is amplified near the particle surface. Plasmonic effects can enhance the photochemistry of silver particles alone or in conjunction with semiconductors according to several mechanisms. We study the photooxidation of citrate by silver nanoparticles in a photoelectrochemical cell, focusing on the wavelength-dependence of the reaction rate and the role of the semiconductor substrate. We find that the citrate photooxidation rate does not track the plasmon resonance of the silver

  2. The extracellular matrix is an integrated unit: ultrastructural localization of collagen types I, III, IV, V, VI, fibronectin, and laminin in human term placenta.

    Science.gov (United States)

    Amenta, P S; Gay, S; Vaheri, A; Martinez-Hernandez, A

    1986-06-01

    The human term placenta is used extensively as a source of extracellular matrix components. To elucidate the tissue distribution and interrelationships of seven of these components, monospecific antibodies directed against collagen types I, III, IV, V, VI, fibronectin, and laminin were reacted with human term placenta and studied by light and electron immunohistochemistry. Type I collagen was the basic structural unit of human term placenta, present as 30-35 nm, cross-banded fibers, often in the form of large fiber bundles. Type III collagen was present as thin 10-15 nm, beaded fibers often forming a meshwork which encased type I collagen fibers. Types V and VI collagen were present as 6-10 nm filaments, often closely associated with types I and III collagen. Type VI collagen also coated collagen fibers of all diameters, enhancing their periodicity, providing a staining pattern often similar to that observed with anti-fibronectin antibodies. Fibronectin was present in both maternal and fetal plasma and throughout the stroma of the chorionic villus, as both free filaments and coating collagen fibers. Basement membranes contained laminin and type IV collagen, but no fibronectin. In summary, the non-basement membrane proteins studied often codistributed with type I collagen, between and apparently attached to fibers, suggesting that they may act as binding proteins, linking type I fibers and bundles, to themselves and to other structures.

  3. Detection of staphylococcal cassette chromosome mec type XI carrying highly divergent mecA, mecI, mecR1, blaZ, and ccr genes in human clinical isolates of clonal complex 130 methicillin-resistant Staphylococcus aureus.

    LENUS (Irish Health Repository)

    Shore, Anna C

    2011-08-01

    Methicillin resistance in staphylococci is mediated by penicillin binding protein 2a (PBP 2a), encoded by mecA on mobile staphylococcal cassette chromosome mec (SCCmec) elements. In this study, two clonal complex 130 (CC130) methicillin-resistant Staphylococcus aureus (MRSA) isolates from patients in Irish hospitals were identified that were phenotypically PBP 2a positive but lacked mecA by conventional PCR and by DNA microarray screening. The isolates were identified as methicillin-susceptible S. aureus using the GeneXpert real-time PCR assay. Whole-genome sequencing of one isolate (M10\\/0061) revealed a 30-kb SCCmec element encoding a class E mec complex with highly divergent blaZ-mecA-mecR1-mecI, a type 8 cassette chromosome recombinase (ccr) complex consisting of ccrA1-ccrB3, an arsenic resistance operon, and flanking direct repeats (DRs). The SCCmec element was almost identical to that of SCCmec type XI (SCCmec XI) identified by the Sanger Institute in sequence type 425 bovine MRSA strain LGA251 listed on the website of the International Working Group on the Classification of Staphylococcal Cassette Chromosome Elements. The open reading frames (ORFs) identified within SCCmec XI of M10\\/0061 exhibited 21 to 93% amino acid identity to ORFs in GenBank. A third DR was identified ca. 3 kb downstream of SCCmec XI, indicating the presence of a possible SCC remnant. SCCmec XI was also identified in the second CC130 MRSA isolate by PCR and sequencing. The CC130 MRSA isolates may be of animal origin as previously reported CC130 S. aureus strains were predominantly from bovine sources. The highly divergent nature of SCCmec XI relative to other SCCmec elements indicates that it may have originated in another taxon.

  4. Sorption behaviour of nanocrystalline MOR type zeolite for Th(IV) and Eu(III) removal from aqueous waste by batch treatment.

    Science.gov (United States)

    Sharma, Pankaj; Tomar, Radha

    2011-10-01

    The nanocrystalline mordenite (MOR) type zeolite materials with initial chemical composition Na(2)O:Al(2)O(3):10SiO(2):48H(2)O have been synthesized under hydrothermal conditions. MOR1 and MOR2 are spherically shaped nanocrystals, whereas MOR3 and MOR4 have rod-like morphology. This paper reports the sorption characteristics of MOR analogues for Th(IV) and Eu(III) removal from aqueous nuclear waste. Sorption of Th(IV) and Eu(III) on MOR1, MOR2, MOR3 and MOR4 in a single component system with varying initial metal ion concentration, solution pH, contact times, sorbent dose and temperatures has also been investigated. Further, the Langmuir and Freundlich sorption models have been applied to describe equilibrium isotherms at different temperatures. The adsorption capacity increases largely with increasing solution pH and temperature of the system. Specific surface area and pore volume have been investigated by Brunauer-Emmett-Teller (BET) method. The N(2) adsorption isotherm presents a type IV isotherm with narrow hysteresis loop which indicates the presence of mesopores related to inter-particle voids. Thermodynamic results indicate that the sorption follows an endothermic physisorption process. It has been found that these exchangers have good sorption capacity and out of which MOR4 has highest sorption capacity. Thus, nanocrystalline MOR4 is proved to be good sorbent for both Th(IV) and Eu(III). Copyright © 2011 Elsevier Inc. All rights reserved.

  5. Type IV Pilus Expression Is Upregulated in Nontypeable Haemophilus influenzae Biofilms Formed at the Temperature of the Human Nasopharynx.

    Science.gov (United States)

    Mokrzan, Elaine M; Ward, Michael O; Bakaletz, Lauren O

    2016-10-01

    Nontypeable Haemophilus influenzae (NTHI), a commensal of the human nasopharynx (hNP), is a common cause of biofilm-associated diseases of the respiratory tract. However, NTHI biofilm biology at the average hNP temperature, i.e., 34°C, has not been well studied. Here we grew NTHI biofilms at 34°C and 37°C, to evaluate relative biofilm growth, expression, and function of the type IV pilus (Tfp), a critical adhesin important for NTHI biofilm formation. The kinetics and regulation of Tfp expression in NTHI biofilms are unclear, especially at 34°C. Tfp expression, as estimated by pilA promoter activity, was distributed throughout the biofilms, with a unique pattern that was dependent on temperature, time in culture, and position within the maturing biofilm. Tfp expression was required for the formation of the characteristic tower structures of NTHI biofilms and was significantly upregulated in NTHI biofilms formed at 34°C versus 37°C. This increase correlated with significantly greater twitching motility at 34°C than at 37°C. Treatment with antisera targeting the major subunit of Tfp (PilA) significantly inhibited NTHI biofilm formation at both temperatures, confirming the importance of this critical adhesin in biofilm formation. Additionally, treatment of preestablished biofilms with antisera against PilA significantly decreased biofilm biomass and mean thickness at both temperatures. These results demonstrated a pivotal role for Tfp in NTHI biofilm formation and stability at the temperature of the hNP, and they underscore the utility of PilA as a vaccine candidate for treatment and/or prevention of NTHI biofilm-associated diseases. NTHI is an important cause of chronic respiratory tract infections, including otitis media, chronic rhinosinusitis, and exacerbations of chronic obstructive pulmonary disease and cystic fibrosis. The chronic and recurrent nature of these diseases is attributed to the presence of bacterial biofilms, which are highly resistant to

  6. [Spa types and antibiotic resistance of Staphylococcus aureus bloodstream isolates obtained form patients of the University Clinical Center in Gdańsk].

    Science.gov (United States)

    Wiśniewska, Katarzyna; Kasprzyk, Joanna; Piechowicz, Lidia; Bronk, Marek; Swieć, Krystyna

    2013-01-01

    Staphylococcus aureus is a leading cause of bloodstream infections. For epidemiological investigations of this bacteria spa genotyping is used as the method which has a high discriminatory power and gives results that can be easily compared between laboratories. In contrast to methicillin-resistant S.aureus (MRSA), relatively little is known about spa types among methicillin-susceptible strains (MSSA). We used spa typing and antibiotic resistance patterns analysis for retrospective study of S.aureus bloodstream isolates population from the University Clinical Centre (UCC) in Gdańsk. The study was performed on 53 isolates from patients of 19 different units/ departments of the UCC. The isolates were tested for the susceptibility to antimicrobial agents. Spa typing was performed on the basis of the sequence analysis of the polymorphic X region of the protein A gene (spa) amplified form the isolates. Spa types were determined by Ridom Staph Type software and were clustered into spa-CCs (clonal complexes) using the algorithm BURP-based upon repeat pattern. MLST (Multilocus Sequence Typing) clonal complexes were predicted from BURP analysis by the Ridom SpaServer database. In MRSA the staphylococcal chromosomal casette (SCC) mec was determined, Spa-typing yielded 26 types. Six spa-CC and seven singletons were identified. The most frequent was spa-CC021involving 38% of isolates. The CC021 consisted of 7 spa types and the most common was t021 corresponding with MLST-CC30. The second frequent was singleton, related to MLST-CC1, with only one type t127. There were 3 MRSA isolates in the population. The MRSA strains were identified as different spa types: t003/ SCCmecII, t008/SCCmecIV and clonally related to MSSA t032/SCCmecIV. No one MRSA strains belonged to spa-CC021. The spa clonal cluster corresponding with widely distributed among invasive S.aureus strains in Europe MLST-CC30 was found as the most frequent among S.aureus bloodstream isolates from the UCC. Occurrence of

  7. Two novel trivacant Keggin-type polytungstates supported manganese carbonyl derivatives synthesized by degradation of metastable [γ-XW10O36]8- (X = Ge(IV), Si(IV)).

    Science.gov (United States)

    Zhao, Juan; Wang, Junsheng; Zhao, Junwei; Ma, Pengtao; Wang, Jingping; Niu, Jingyang

    2012-05-21

    Two novel trivacant Keggin-type polyoxometallate-based manganese carbonyl derivatives K(8)[(OC)(3)Mn(A-α-H(2)GeW(9)O(34))]·10H(2)O (1) and K(8)[(OC)(3)Mn(A-α-H(2)SiW(9)O(34))]·11H(2)O (2) have been synthesized by degradation of the metastable [γ-XW(10)O(36)](8-) (X = Ge(IV), Si(IV)) in CH(3)CN-H(2)O solvent (1 : 2, vol.) and structurally characterized by elemental analyses, IR spectra, thermogravimetric analysis, and single-crystal X-ray diffraction. X-ray diffraction analyses indicate that 1 and 2 are isomorphic and consist of a [(OC)(3)Mn](+) group and a trivacant [A-α-H(2)XW(9)O(34)](8-) fragment, which exhibit the first examples of trivacant Keggin-type metal carbonyl derivatives. Additionally, the electrocatalytic properties of both for NO(2)(-) oxidation have been investigated.

  8. CagI is an essential component of the Helicobacter pylori Cag type IV secretion system and forms a complex with CagL.

    Directory of Open Access Journals (Sweden)

    Kieu Thuy Pham

    Full Text Available Helicobacter pylori, the causative agent of type B gastritis, peptic ulcers, gastric adenocarcinoma and MALT lymphoma, uses the Cag type IV secretion system to induce a strong proinflammatory response in the gastric mucosa and to inject its effector protein CagA into gastric cells. CagA translocation results in altered host cell gene expression profiles and cytoskeletal rearrangements, and it is considered as a major bacterial virulence trait. Recently, it has been shown that binding of the type IV secretion apparatus to integrin receptors on target cells is a crucial step in the translocation process. Several bacterial proteins, including the Cag-specific components CagL and CagI, have been involved in this interaction. Here, we have examined the localization and interactions of CagI in the bacterial cell. Since the cagI gene overlaps and is co-transcribed with the cagL gene, the role of CagI for type IV secretion system function has been difficult to assess, and conflicting results have been reported regarding its involvement in the proinflammatory response. Using a marker-free gene deletion approach and genetic complementation, we show now that CagI is an essential component of the Cag type IV secretion apparatus for both CagA translocation and interleukin-8 induction. CagI is distributed over soluble and membrane-associated pools and seems to be partly surface-exposed. Deletion of several genes encoding essential Cag components has an impact on protein levels of CagI and CagL, suggesting that both proteins require partial assembly of the secretion apparatus. Finally, we show by co-immunoprecipitation that CagI and CagL interact with each other. Taken together, our results indicate that CagI and CagL form a functional complex which is formed at a late stage of secretion apparatus assembly.

  9. Immunohistochemical assessment of collagen types I, III, IV and VI in biopsy samples of the bovine uterine wall collected during the oestrous cycle.

    Science.gov (United States)

    Boos, A

    2000-01-01

    Uterine biopsies were collected at cycle days 1 (oestrous), 8, 15 and 19 in six cows. Unfixed cryostat sections were used to immunolocalise collagen types I, III, IV and VI by an indirect FITC method. Collagen I was sparsely found in the endometrium where it formed a fine meshwork of thin fibres directly below the surface epithelium, clearly visible only at cycle days 8 and 15. Collagen III formed the bulk of connective tissue fibres and was arranged in fine aggregates within the superficial endometrial stroma, while in the deeper areas it consisted of many thick fibre bundles. Collagen IV was found in basement membranes underlying all endometrial epithelia. Furthermore, it surrounded smooth muscle cells of blood vessels. A few single fibrils also stained positively within the endometrial stroma, more numerous at cycle days 1 and 19 as compared to days 8 and 15. Collagen VI formed a mesh of fine and pericellularly situated fibrils within the endometrial stroma. The contribution of the collagen types studied to the connective tissue of caruncles, blood vessels, lymph follicles, and myometrium is also reported. The results of the present study indicate that the connective tissue of the bovine uterine wall is composed of different collagen types, which exhibit a characteristic distribution pattern each. The day of cycle may influence amounts and organisation of collagen types I and IV as demonstrated here at the light-microscopical level. Copyright 2000 S. Karger AG, Basel

  10. Distribution of basal lamina type IV collagen and laminin in normal rat tongue mucosa and experimental oral carcinoma: ultrastructural immunolocalization and immunogold quantitation.

    Science.gov (United States)

    Jiang, D J; Wilson, D F; Smith, P S; Pierce, A M; Wiebkin, O W

    1994-07-01

    The relationship of basal lamina, a form of specialised extracellular matrix which separates epithelial cells and other cell types from adjacent stroma, to the behaviour of malignant neoplasms of epithelial origin is not well understood. However, it is widely acknowledged that the properties of local invasion and metastasis of carcinomas are linked to extracellular matrix (including basal lamina) changes. In the present study, the distribution of the major basal lamina components, type IV collagen and laminin, in normal rat tongue mucosa and experimentally induced oral carcinomas was investigated using post-embedding immunogold techniques and electron microscopy. The expression of these components was also quantitatively analysed using morphometry and immunocytochemistry. Results indicated that type IV collagen and laminin were confined to the lamina densa of normal oral epithelial basal lamina, and that both components were also detected in the lamina densa of basal lamina associated with carcinomas, and in the extracellular matrix of tumours. Furthermore, laminin was detected within stromal fibroblasts in normal tissues and experimental carcinomas. Quantitative analysis indicated that expression of laminin was significantly increased in carcinomas. In contrast, type IV collagen expression was significantly decreased. The quantitative changes observed in the two basal lamina constituents may be related to the process of tumour invasion, reflecting altered metabolic activities of tumour and stromal cells. These observations may be of use in understanding the architectural characteristics of oral mucosa basal lamina and in assessing the malignant potential of epithelial dysplasias or "premalignant" lesions.

  11. The dipeptidyl peptidase IV inhibitor vildagliptin suppresses endogenous glucose production and enhances islet function after single-dose administration in type 2 diabetic patients

    DEFF Research Database (Denmark)

    Balas, Bogdan; Baig, Muhammad R; Watson, Catherine

    2007-01-01

    AIMS/HYPOTHESIS: Vildagliptin is a selective dipeptidyl peptidase IV inhibitor that augments meal-stimulated levels of biologically active glucagon-like peptide-1. Chronic vildagliptin treatment decreases postprandial glucose levels and reduces hemoglobin A1c in type 2 diabetic patients. However......, little is known about the mechanism(s) by which vildagliptin promotes reduction in plasma glucose concentration. METHODS: Sixteen patients with type 2 diabetes (age, 48+/-3 yr; body mass index, 34.4+/-1.7 kg/m2; hemoglobin A1c, 9.0+/-0.3%) participated in a randomized, double-blind, placebo......-controlled trial. On separate days patients received 100 mg vildagliptin or placebo at 1730 h followed 30 min later by a meal tolerance test (MTT) performed with double tracer technique (3-(3)H-glucose iv and 1-(14)C-glucose orally). RESULTS: After vildagliptin, suppression of endogenous glucose production (EGP...

  12. Modification of Pseudomonas aeruginosa Pa5196 Type IV Pilins at Multiple Sites with d-Araf by a Novel GT-C Family Arabinosyltransferase, TfpW▿

    OpenAIRE

    Kus, Julianne V.; Kelly, John; Tessier, Luc; Harvey, Hanjeong; Cvitkovitch, Dennis G.; Burrows, Lori L.

    2008-01-01

    Pseudomonas aeruginosa Pa5196 produces type IV pilins modified with unusual α1,5-linked d-arabinofuranose (α1,5-d-Araf) glycans, identical to those in the lipoarabinomannan and arabinogalactan cell wall polymers from Mycobacterium spp. In this work, we identify a second strain of P. aeruginosa, PA7, capable of expressing arabinosylated pilins and use a combination of site-directed mutagenesis, electrospray ionization mass spectrometry (MS), and electron transfer dissociation MS to identify th...

  13. Glycosylation of Pseudomonas aeruginosa Strain Pa5196 Type IV Pilins with Mycobacterium-Like α-1,5-Linked d-Araf Oligosaccharides▿

    OpenAIRE

    Voisin, Sébastien; Kus, Julianne V.; Houliston, Scott; St-Michael, Frank; Watson, Dave; Cvitkovitch, Dennis G.; Kelly, John; Brisson, Jean-Robert; Burrows, Lori L.

    2006-01-01

    Pseudomonas aeruginosa is a gram-negative bacterium that uses polar type IV pili for adherence to various materials and for rapid colonization of surfaces via twitching motility. Within the P. aeruginosa species, five distinct alleles encoding variants of the structural subunit PilA varying in amino acid sequence, length, and presence of posttranslational modifications have been identified. In this work, a combination of mass spectrometry and nuclear magnetic resonance spectroscopy was used t...

  14. Collagen types I, III and IV in the placentome and interplacentomal maternal and fetal tissues in normal cows and in cattle with retention of fetal membranes.

    Science.gov (United States)

    Boos, A; Stelljes, A; Kohtes, J

    2003-01-01

    The increase in uterine mass during pregnancy requires the establishment of sufficient blood supply to and strong supportive elements within the uterus. These needs are correlated with the remodelling and production of ECM materials. Therefore, placentomes and interplacentomal parts of the uterine walls and adherent allantochorion were collected from 45 cows at slaughter. Additional placentomes were obtained from 5 cows at premature cesarean section and at term in 5 cows releasing their fetal membranes in time or in 5 animals with retention of the fetal membranes, i.e. in total 60 pregnancies. Unfixed cryostat sections from 4 animals per month of pregnancy and 5 animals per peripartal group (in total 51 pregnancies) were used to immunolocalize collagen types I, III, and IV by an indirect FITC method. Collagen types I and III co-localize within the uterus. The tensile strength of the pregnant uterus is mainly represented by high contents of collagen type I within the allantochorion and subepithelial endometrial and subserosal meshes. Chorionic villi are fixed within caruncular crypts by two mechanisms: crypt openings are narrow and supplied with thick edges containing collagen types I and III. Collagen type IV contributes to all basement membranes and encloses connective tissue cells within the maternal crypt stroma, the stratum compactum and the perimetrial connective tissue. At term, fetal membranes and placentomes are edematous and at the light-microscopic level no distinct differences are visible between connective tissue fibers of placentomes from animals retaining the fetal membranes and those releasing them in time. In conclusion, collagen types I, III and IV exhibit type- and location-specific distribution patterns within the uterus of the pregnant cow. These may additionally be influenced by the stage of pregnancy, thus reflecting the dynamic processes at the stromal level. Copyright 2003 S. Karger AG, Basel

  15. Endovenous surgery for recurrent varicose veins with a one-year follow up in a patient with Ehlers Danlos syndrome type IV.

    Science.gov (United States)

    Whiteley, Mark S; Holdstock, Judith M

    2015-08-01

    We present a woman with severe symptomatic recurrent varicose veins who was treated with endovenous laser ablation and transluminal occlusion of perforator with attempted phlebectomies for extensive varices. The phlebectomies turned out to be near impossible due to friability of the veins. Her treatment was completed with post-operative ultrasound guided foam sclerotherapy seven months later. She was subsequently diagnosed as Ehlers Danlos syndrome type IV. A duplex ultrasound scan 18 months post-endovenous laser ablation and transluminal occlusion of perforator and 11 months after ultrasound guided foam sclerotherapy confirmed successful closure with virtual atrophy of all treated veins. She was found to be reflux free and only showed a few scattered cosmetic reticular veins. Open varicose vein surgery has been reported as being hazardous in the past in a patient with Ehlers Danlos syndrome type IV. Our experience has shown that endovenous laser ablation, transluminal occlusion of perforator and ultrasound guided foam sclerotherapy appear to be effective in treating this patient with Ehlers Danlos syndrome type IV, although phlebectomies were technically impossible. © The Author(s) 2014.

  16. Temporal and spatial expression of laminin, collagen types IV and I and alpha 6/beta 1 integrin receptor in the developing rat parotid gland.

    Science.gov (United States)

    Lazowski, K W; Mertz, P M; Redman, R S; Kousvelari, E

    1994-04-01

    We have examined the temporal expression and cellular localization of the genes and proteins for the extracellular matrix (ECM) proteins laminin (B1, B2 and A chain), collagen types alpha 1 (IV) and alpha 1 (I) and the integrin receptor complex alpha 6/beta 1, during parotid gland postnatal development. Laminin B1 and B2 isoforms and collagens alpha 1 (IV) and alpha 1 (I) mRNA steady-state levels were highest at ages 0, 7 and 14 days after birth and declined to the adult (90 days) level at 21 days and older. Laminin A chain transcripts were not detected at any age. Collagen alpha 1 (IV) and laminin were localized in the basal membrane of the developing acinar and ductal cells, while collagen alpha 1 (I) was localized in the stroma surrounding the cells. The amounts of these ECM components were high at the early stages of development and lower at later times. The pattern of expression of the alpha 6/beta 1 integrin genes during development was similar to those of laminin and collagens alpha 1 (IV) and alpha 1 (I). Accumulations of mRNA were high at 0, 7 and 14 days after birth and lower at 21 days and older. High levels of beta 1 integrin were localized in the developing acinar and ductal cell membranes at early ages (7 days); lower amounts were present in the same distribution pattern at later stages of gland development.(ABSTRACT TRUNCATED AT 250 WORDS)

  17. Synthesis and in vitro anticancer activity of octahedral platinum(IV) complexes with cyclohexyl-functionalized ethylenediamine-N,N'-diacetate-type ligands.

    Science.gov (United States)

    Lazić, Jelena M; Vucićević, Ljubica; Grgurić-Sipka, Sanja; Janjetović, Kristina; Kaluderović, Goran N; Misirkić, Maja; Gruden-Pavlović, Maja; Popadić, Dusan; Paschke, Reinhard; Trajković, Vladimir; Sabo, Tibor J

    2010-06-07

    The present study describes the synthesis and anticancer activity of novel octahedral Pt(IV) complexes with cyclohexyl functionalized ethylenediamine-N,N'-diacetate-type ligands. Molecular mechanics calculations and density functional theory analysis revealed that s-cis is the preferred geometry of these Pt(IV) complexes with tetradentate-coordinated (S,S)-ethylenediamine-N,N'-di-2-(3-cyclohexyl)propanoate. The viability of cancer cell lines (U251 human glioma, C6 rat glioma, L929 mouse fibrosarcoma, and B16 human melanoma) was assessed by measuring mitochondrial dehydrogenase activity and lactate dehydrogenase release. Cell-cycle distribution, oxidative stress, caspase activation, and induction of autophagy were analyzed by flow cytometry using appropriate fluorescent reporter dyes. The cytotoxic activity of novel Pt(IV) complexes against various cancer cell lines (IC(50) range: 1.9-8.7 microM) was higher than that of cisplatin (IC(50) range: 10.9-67.0 microM) and proceeded through completely different mechanisms. Cisplatin induced caspase-dependent apoptosis associated with the cytoprotective autophagic response. In contrast, the new Pt(IV) complexes caused rapid, caspase-independent, oxidative stress-mediated non-apoptotic cell death characterized by massive cytoplasmic vacuolization, cell membrane damage, and the absence of protective autophagy.

  18. Capmatinib, Ceritinib, Regorafenib, or Entrectinib in Treating Patients With BRAF/NRAS Wild-Type Stage III-IV Melanoma

    Science.gov (United States)

    2017-12-20

    ALK Fusion Protein Expression; BRAF wt Allele; Invasive Skin Melanoma; MET Fusion Gene Positive; NRAS wt Allele; NTRK1 Fusion Positive; NTRK2 Fusion Positive; NTRK3 Fusion Positive; RET Fusion Positive; ROS1 Fusion Positive; Stage III Cutaneous Melanoma AJCC v7; Stage IIIA Cutaneous Melanoma AJCC v7; Stage IIIB Cutaneous Melanoma AJCC v7; Stage IIIC Cutaneous Melanoma AJCC v7; Stage IV Cutaneous Melanoma AJCC v6 and v7

  19. Transmission of endemic ST22-MRSA-IV on four acute hospital wards investigated using a combination of spa, dru and pulsed-field gel electrophoresis typing.

    LENUS (Irish Health Repository)

    Creamer, E

    2012-11-01

    The transmission of meticillin-resistant Staphylococcus aureus (MRSA) between individual patients is difficult to track in institutions where MRSA is endemic. We investigated the transmission of MRSA where ST22-MRSA-IV is endemic on four wards using demographic data, patient and environmental screening, and molecular typing of isolates. A total of 939 patients were screened, 636 within 72 h of admission (on admission) and 303 >72 h after admission, and 1,252 environmental samples were obtained. Isolates were typed by spa, dru and pulsed-field gel electrophoresis (PFGE) typing. A composite dendrogram generated from the three sets of typing data was used to divide isolates into \\'dendrogram groups\\' (DGs). Ten percent of patients (92\\/939) were MRSA-positive; 7 % (44\\/636) on admission and 16 % (48\\/303) >72 h after admission (p = 0.0007). MRSA was recovered from 5 % of environmental specimens (65\\/1,252). Most isolates from patients (97 %, 85\\/88) and the environment (97 %, 63\\/65) exhibited the ST22-MRSA-IV genotype. Four DGs (DG1, DG4, DG16 and DG17) accounted for 58 % of ST22-MRSA-IV isolates from patients. Epidemiological evidence suggested cross-transmission among 44\\/92 patients (48 %) but molecular typing confirmed probable cross-transmission in only 11 instances (13 %, 11\\/88), with the majority of cross-transmission (64 %; 7\\/11) occurring on one ward. In the setting of highly clonal endemic MRSA, the combination of local epidemiology, PFGE, spa and dru typing provided valuable insights into MRSA transmission.

  20. Liver fibrosis in elderly cadavers: localization of collagen types I, III, and IV, α-smooth muscle actin, and elastic fibers.

    Science.gov (United States)

    Mak, Ki M; Chu, Edward; Lau, K H Vincent; Kwong, Allison J

    2012-07-01

    We have shown a high prevalence of liver fibrosis in elderly cadavers with diverse causes of death by Sirius red stain; however, the various collagen types in these samples have yet to be evaluated. To further characterize the histopathology of the fibrotic lesions in the livers of these elderly cadavers, this study used immunohistochemistry and histochemistry to identify the principal collagens produced in liver fibrosis, fibrogenic cells and elastic fibers. Collagen I and III immunoreactions were found to colocalize in collagen fibers of fibrotic central veins, perisinusoidal fibrotic foci, portal tract stroma, and fibrous septa. α-Smooth muscle actin-expressing perisinusoidal hepatic stellate cells (HSCs), as well as perivenular, portal, and septal myofibroblasts, were closely associated with collagen fibers, reflecting their fibrogenic functions. HSCs and myofibroblasts were also noted to express collagen IV, which may contribute to production of basal lamina-like structures. In fibrotic livers, the sinusoidal lining showed variable immunostaining for collagen IV. Collagen IV immunostaining revealed vascular proliferation and atypical ductular reaction at the portal-septal parenchymal borders, as well as capillary-like vessels in the lobular parenchyma. While elastic fibers were absent in the space of Disse, they were found to codistribute with collagens in portal tracts, fibrous septa and central veins. Our combined assessment of collagen types, HSCs, myofibroblasts, and elastic fibers is significant in understanding the histopathology of fibrosis in the aging liver. Copyright © 2012 Wiley Periodicals, Inc.

  1. A catalogue of the Coelenterate type specimens of the Zoological Museum of Amsterdam. IV. Gorgonacea, Actiniaria, Scleractinia

    NARCIS (Netherlands)

    Soest, van R.W.M.

    1979-01-01

    This final part of the Coelenterate type catalogue lists 274 nominal types (183 Gorgonacea, 9 Actiniaria and 82 Scleractinia). Six not previously depicted types are represented by photographic illustrations of their habit. Furthermore lists are provided of schizotypes, i.e. fragments of types housed

  2. [Immunohistochemical studies of collagen types I, III and IV distribution and the expression of procollagen III peptide in papillary carcinomas of the thyroid].

    Science.gov (United States)

    Katoh, R; Ono, S; Sasaki, J; Takayama, K; Nishio, H; Tomichi, N; Yagawa, K

    1990-08-01

    Collagen types I, III and IV and procollagen III peptide (P III P) were examined immunohistochemically in 38 papillary carcinomas of the thyroid. The immunoreactivity to both type I and type III collagen was diffuse and abundantly observed in the fibrous stroma, where it displayed a fibrillar and/or granular pattern with little difference in the intensity of the staining. The immunoreactivity to collagen type IV was localized in the basement membrane of the vessels and partly in the tumor cell nests. A distinctive cytoplasmic immunoreactivity to P III P was observed in the cancer cells and the fibroblasts in all of the 38 papillary carcinomas, specifically an intense and extensive immunoreactivity by the cancer cells present in tumors having fibrosclerosing stroma and/or showing an extensive local invasion. Thus, it is conceivable that stromal collagen in a papillary carcinoma can be produced by not only the fibroblasts but also by tumor cells and its productivity could be affected by the degree of the tumoral invasion.

  3. Linear dimensional change, compressive strength and detail reproduction in type IV dental stone dried at room temperature and in a microwave oven.

    Science.gov (United States)

    Silva, Marcos Aurélio Bomfim da; Vitti, Rafael Pino; Consani, Simonides; Sinhoreti, Mário Alexandre Coelho; Mesquita, Marcelo Ferraz; Consani, Rafael Leonardo Xediek

    2012-01-01

    The type IV dental stone is widely used for the fabrication of dyes and master casts for fixed and removable partial prostheses. It is typically normal to wait at least 24 hours for the casts to dry prior to beginning the laboratory procedures. The waiting time has been shown to be greatly reduced by using microwave drying. This study evaluated the influence of drying techniques at room temperature and microwave oven on the linear dimensional change, compressive strength and detail reproduction in type IV dental stones. Three type IV dental stone brands were selected; elite Rock, Shera Premium and Durone IV. Two different drying protocols were tested in 4 groups (n=10); G-room temperature (25±4 ºC) dried for 2 hours; G2--room temperature dried for 24 hours; G3-room temperature dried for 7 days and G4--microwave oven dried at 800 W for 5 minutes and after 2 hours at room temperature. After drying, the samples were assayed for dimensional charges. The sample surface was submitted to the ImageTool 3.0 software for compressive strength in a universal testing machine with a cell load of 50 KN at a crosshead speed of 0.5 mm/minutes and the detail reproduction was analyzed with a stereomicroscope at 25x magnification. The statistical analysis of the linear dimensional change and compressive strength data were conducted by the ANOVA test followed by the Tukey test (pmicrowave oven drying showed a linear dimensional change similar to after room temperature drying for 24 hours and 7 days. The compressive strength of the stone dried in the microwave oven was similar to those dried at room temperature for 24 hours, with the exception of Shera Premium, which had similar results for microwave and room temperature drying for 7 days. For the microwave drying method the detail reproduction levels for samples dried at room temperature for 24 hours and 7 days were similar, except for the Durone IV.

  4. Mechanisms and implications of a type IV functional response for short-term intake rate of dry matter in large mammalian herbivores.

    Science.gov (United States)

    Mezzalira, Jean C; Bonnet, Olivier J F; Carvalho, Paulo C de F; Fonseca, Lidiane; Bremm, Carolina; Mezzalira, Carlos C; Laca, Emilio A

    2017-09-01

    The functional response (i.e. the relationship between consumers' intake rate and resource density) is central in plant-herbivore interactions. Its shape and the biological processes leading to it have significant implications for both foraging theory and ecology of grazing systems. A type IV functional response (i.e. dome-shaped relationship) of short-term intake rate of dry matter (intake while grazing) has rarely been reported for large herbivores and the conditions that can lead to it are poorly understood. We report a type IV functional response observed in heifers grazing monocultures of Cynodon sp. and Avena strigosa. The mechanisms and consequences of this type of functional response for grazed system dynamics are discussed. Intake rate was higher at intermediate than at short or tall sward heights in both grass species. The type IV functional response resulted from changes in bite mass instead of a longer time needed to encounter and process bites. Thus, the decrease of intake rate of dry matter in tall swards is not explained by a shift from process 3 (potential bites are concentrated and apparent) to process 2 (potential bites are apparent but dispersed, Spalinger & Hobbs 1992). Bite mass was smaller in tall than in intermediate swards due to a reduction of bite volume possibly caused by the greater proportion of stem and sheath acting as a physical barrier to bite formation. It is generally accepted that potential bites are abundant and apparent in most grassland and meadow systems, as they were in the present experiments. Therefore, a type IV response of intake rate not directly related to digestive constraints may determine the dynamics of intake and defoliation under a much larger set of conditions than previously thought. These results have implications for foraging theory and stability of grazing systems. For example, if animals prefer patches of intermediate stature that yield the highest intake rate, grazing should lead to the widely observed

  5. Functional Live-Cell Imaging Demonstrates that β1-Integrin Promotes Type IV Collagen Degradation by Breast and Prostate Cancer Cells

    Directory of Open Access Journals (Sweden)

    Mansoureh Sameni

    2008-09-01

    Full Text Available The ability of tumor cells to adhere to, migrate on, and remodel extracellular matrices is mediated by cell surface receptors such as β1-integrins. Here we conducted functional live-cell imaging in real time to investigate the effects of modulating β1-integrin expression and function on proteolytic remodeling of the extracellular matrix. Human breast and prostate cancer cells were grown on reconstituted basement membrane containing a quenched fluorescent form of collagen IV. Generation of cleavage products and the resulting increases in fluorescence were imaged and quantified. Decreases in the expression and activity of β1-integrin reduced digestion of quenched fluorescent-collagen IV by the breast and prostate cancer cells and correspondingly their invasion through and migration on reconstituted basement membrane. Decreased extracellular matrix degradation also was associated with changes in the constituents of proteolytic pathways: decreases in secretion of the cysteine protease cathepsin B, the matrix metalloproteinase (MMP-13, and tissue inhibitors of metalloproteinases (TIMP-1 and 2; a decrease in expression of MMP-14 or membrane type 1 MMP; and an increase in secretion of TIMP-3. This is the first study to demonstrate through functional live-cell imaging that downregulation of β1-integrin expression and function reduces proteolysis of collagen IV by breast and prostate cancer cells.

  6. Genetic and mechanistic evaluation for the weak A phenotype in Ael blood type with IVS6 + 5G>A ABO gene mutation.

    Science.gov (United States)

    Chen, D-P; Sun, C-F; Ning, H-C; Peng, C-T; Wang, W-T; Tseng, C-P

    2015-01-01

    Ael is a rare blood type that is characterized by weak agglutination of RBCs when reacts with anti-A antibody in adsorption-elution test. Although IVS6 + 5G→A mutation is known to associate with the Ael blood type, genetic and mechanistic evaluation for the weak agglutination of Ael with IVS6 + 5G→A mutation has not yet been completely addressed. In this study, five cases of confirmed Ael individuals were analysed. The cDNAs for the A(el) alleles were obtained by cloning method for sequence analyses. The erythroleukemia K562 cells were used as the cell study model and were transfected with the A(el) expression construct. Flow cytometry analysis was then performed to determine the levels of surface antigen expression. The results indicated that IVS6 + 5G→A attributes to all cases of Ael . RT-PCR analyses revealed the presence of at least 10 types of aberrant A(el) splicing transcripts. Most of the transcripts caused early termination and produced non-functional protein during translation. Nevertheless, the transcript without exons 5-6 was predicted to generate functional Ael glycosyltransferase lacking 57 amino acids at the N-terminal segment. When the exons 5-6 deletion transcript was stably expressed in the K562 cells, weak agglutination of the cells can be induced by adding anti-A antibody followed by adsorption-elution test. This study demonstrates that aberrant splicing of A transcripts contributes to weak A expression and the weak agglutination of Ael -RBCs, adding to the complexity for the regulatory mechanisms of ABO gene expression. © 2014 International Society of Blood Transfusion.

  7. Wolbachia lipoproteins: abundance, localisation and serology of Wolbachia peptidoglycan associated lipoprotein and the Type IV Secretion System component, VirB6 from Brugia malayi and Aedes albopictus.

    Science.gov (United States)

    Voronin, Denis; Guimarães, Ana F; Molyneux, Gemma R; Johnston, Kelly L; Ford, Louise; Taylor, Mark J

    2014-10-06

    Lipoproteins are the major agonists of Wolbachia-dependent inflammatory pathogenesis in filariasis and a validated target for drug discovery. Here we characterise the abundance, localisation and serology of the Wolbachia lipoproteins: Wolbachia peptidoglycan associated lipoprotein and the Type IV Secretion System component, VirB6. We used proteomics to confirm lipoprotein presence and relative abundance; fractionation, immunoblotting and confocal and electron immuno-microscopy for localisation and ELISA for serological analysis. Proteomic analysis of Brugia malayi adult female protein extracts confirmed the presence of two lipoproteins, previously predicted through bioinformatics: Wolbachia peptidoglycan associated lipoprotein (wBmPAL) and the Type IV Secretion System component, VirB6 (wBmVirB6). wBmPAL was among the most abundant Wolbachia proteins present in an extract of adult female worms with wBmVirB6 only detected at a much lower abundance. This differential abundance was reflected in the immunogold-labelling, which showed wBmPAL localised at numerous sites within the bacterial membranes, whereas wBmVirB6 was present as a single cluster on each bacterial cell and also located within the bacterial membranes. Immunoblotting of fractionated extracts confirmed the localisation of wBmPAL to membranes and its absence from cytosolic fractions of C6/36 mosquito cells infected with wAlbB. In whole worm mounts, antibody labelling of both lipoproteins were associated with Wolbachia. Serological analysis showed that both proteins were immunogenic and raised antibody responses in the majority of individuals infected with Wuchereria bancrofti. Two Wolbachia lipoproteins, wBmPAL and wBmVirB6, are present in extracts of Brugia malayi with wBmPAL among the most abundant of Wolbachia proteins. Both lipoproteins localised to bacterial membranes with wBmVirB6 present as a single cluster suggesting a single Type IV Secretory System on each Wolbachia cell.

  8. A Matched Comparative Study of the Bilhaut Procedure Versus Resection and Reconstruction for Treatment of Radial Polydactyly Types II and IV.

    Science.gov (United States)

    Dijkman, Robert R; Selles, Ruud W; Hülsemann, Wiebke; Mann, Max; Habenicht, Rolf; Hovius, Steven E R; van Nieuwenhoven, Christianne A

    2016-05-01

    To compare outcomes of the Bilhaut procedure with outcomes of conventional resection and reconstruction in radial polydactyly types II and IV. Patients treated with the Bilhaut procedure were radiologically matched with patients treated with reconstruction. Evaluated outcomes included the Rotterdam assessment system, pinch strength, and thumb size measurements. To determine objectively which aesthetic outcome scores truly depended on surgical technique rather than observer opinion, we analyzed evaluations by a panel of 22 individuals with varying clinical experience who were blinded to the study protocol, using a linear mixed regression model. The Bilhaut procedure reduced the risk of suboptimal outcome for metacarpophalangeal joint instability in type IV radial polydactyly. Conversely, the Bilhaut procedure increased the risk of suboptimal scar appearance, residual prominence at amputation site, thumb size, and nail appearance. Tip pinch strength was more significantly reduced after the Bilhaut compared with reconstruction, whereas pulp circumference and nail width exceeded 100% of the unaffected contralateral hand after the Bilhaut reconstruction. There was no significant difference in active range of motion between procedures. Nail appearance proved the only aesthetic drawback of the Bilhaut procedure after adjustment for clinical experience. There was superior metacarpophalangeal joint stability after the Bilhaut procedure for radial polydactyly type IV, but this did not result in the presumed benefit to thumb strength. For experienced surgeons, both procedures resulted in comparable thumb active range of motion. However, aesthetic results were more likely perceived as pleasing after conventional reconstruction, even after adjusting for observer experience with regard to nail appearance. Despite possible benefits of modified Bilhaut procedures that preserve the nail, conventional reconstruction is the preferred procedure until otherwise proven. Therapeutic III

  9. Effect of fibroblast activation protein and alpha2-antiplasmin cleaving enzyme on collagen types I, III, and IV.

    Science.gov (United States)

    Christiansen, Victoria J; Jackson, Kenneth W; Lee, Kyung N; McKee, Patrick A

    2007-01-15

    The circulating enzyme, alpha2-antiplasmin cleaving enzyme (APCE), has very similar sequence homology and proteolytic specificity as fibroblast activation protein (FAP), a membrane-bound proteinase. FAP is expressed on activated fibroblasts associated with rapid tissue growth as in embryogenesis, wound healing, and epithelial-derived malignancies, but not in normal tissues. Its presence on stroma suggests that FAP functions to remodel extracellular matrix (ECM) during neoplastic growth. Precise biologic substrates have not been defined for FAP, although like APCE, it cleaves alpha2-antiplasmin to a derivative more easily cross-linked to fibrin. While FAP has been shown to cleave gelatin, evidence for cleavage of native collagen, the major ECM component, remains indistinct. We examined the potential proteolytic effects of FAP or APCE alone and in concert with selected matrix metalloproteinases (MMPs) on collagens I, III, and IV. SDS-PAGE analyses demonstrated that neither FAP nor APCE cleaves collagen I. Following collagen I cleavage by MMP-1, however, FAP or APCE digested collagen I into smaller peptides. These peptides were analogous to, yet different from, those produced by MMP-9 following MMP-1 cleavage. Amino-terminal sequencing and mass spectrometry analyses of digestion mixtures identified several peptide fragments within the sequences of the two collagen chains. The proteolytic synergy of APCE in the cleavage of collagen I and III was not observed with collagen IV. We conclude that FAP works in synchrony with other proteinases to cleave partially degraded or denatured collagen I and III as ECM is excavated, and that derivative peptides might function to regulate malignant cell growth and motility.

  10. Ehlers-Danlos syndrome type IV and recurrent carotid-cavernous fistula: review of the literature, endovascular approach, technique and difficulties

    Energy Technology Data Exchange (ETDEWEB)

    Desal, H.A.; Toulgoat, F.; Raoul, S.; Guillon, B.; Bommard, S.; Naudou-Giron, E.; Auffary-Calvier, E.; Kersaint-Gilly, A. de [Department of Neuroradiology, Laennec Hospital, University of Nantes (France); 1

    2005-04-01

    We report the follow-up of a previously published case (Forlodou et al. Neuroradiology 38:595-597, 1996) of carotido-cavernous fistulas (CCFs) in a patient presenting with type IV Ehlers-Danlos syndrome (EDS 4) that were successfully treated twice by an endovascular approach. Initial treatment with a detachable balloon was in 1994 for a right CCF, and, 8 years later, a left CCF was treated by selective transarterial occlusion of the cavernous sinus with coils. Unfortunately, the patient suffered from a spontaneous post-operative intracranial haemorrhage in the left hemisphere and died. Review of the literature, technical considerations for bilateral CCF and complication are discussed.

  11. Surgery for Type III-IV hiatal hernia: anatomical recurrence and global results after elective treatment of short oesophagus with open and minimally invasive surgery.

    Science.gov (United States)

    Lugaresi, Marialuisa; Mattioli, Benedetta; Daddi, Niccolò; Di Simone, Massimo Pierluigi; Perrone, Ottorino; Mattioli, Sandro

    2016-04-01

    Type III-IV hiatal hernia (HH) is associated with a true short oesophagus in more than 50% of cases; dedicated treatment of this condition might be appropriate to reduce the recurrence rate after surgery. A case series of patients receiving surgery for Type III-IV hernia was examined for short oesophagus, and the results were analysed. From 1980 to 1994, 60 patients underwent an open surgical approach, and the position of the oesophago-gastric junction was visually localized; from 1995 to 2013, 48 patients underwent a minimally invasive approach, and the oesophago-gastric junction was objectively localized using a laparoscopic-endoscopic method. The patients underwent a timed clinical-instrumental follow-up that included symptoms assessment, barium swallow and endoscopy. The results were considered to be excellent in the absence of symptoms and oesophagitis; good, if symptoms occurred two to four times a month in the absence of oesophagitis; fair, if symptoms occurred two to four times a week in the presence of hyperaemia, oedema and/or microscopic oesophagitis; and poor, if symptoms occurred on a daily basis in the presence of any grade of endoscopic oesophagitis, HH of any size or type, or the need for antireflux medical therapy. The follow-up time was calculated from the time of surgery to the last complete follow-up. Among the open surgery patients, 78% underwent abdominal fundoplication, 10% the Belsey Mark IV procedure, 8% laparotomic Collis-Nissen fundoplication and 3% the Pearson operation. Among the minimally invasive surgery patients, 44% underwent a laparoscopic floppy Nissen procedure and 56% a left thoracoscopic Collis-laparoscopic Nissen procedure. The postoperative mortality and complication rates were 1.6% (1/60) and 15% for open surgery and 4.1% (2/48) and 12.5% for minimally invasive surgery. A total of 105 patients were followed up for a median period of 96 months. Five relapses occurred after open surgery (5/59, 8%) and two after minimally

  12. Brucella abortus VirB12 Is Expressed during Infection but Is Not an Essential Component of the Type IV Secretion System

    OpenAIRE

    Sun, Yao-Hui; Rolán, Hortensia G.; den Hartigh, Andreas B.; Sondervan, David; Tsolis, Renée M.

    2005-01-01

    The Brucella abortus virB operon, consisting of 11 genes, virB1 to virB11, and two putative genes, orf12 (virB12) and orf13, encodes a type IV secretion system (T4SS) that is required for intracellular replication and persistent infection in the mouse model. This study was undertaken to determine whether orf12 (virB12) encodes an essential part of the T4SS apparatus. The virB12 gene was found to encode a 17-kDa protein, which was detected in vitro in B. abortus grown to stationary phase. Mice...

  13. Vildagliptin, a dipeptidyl peptidase-IV inhibitor, improves model-assessed beta-cell function in patients with type 2 diabetes

    DEFF Research Database (Denmark)

    Mari, A; Sallas, W M; He, Y L

    2005-01-01

    AIMS/HYPOTHESIS: The dipeptidyl peptidase IV inhibitor, vildagliptin, increases levels of intact glucagon-like peptide-1 (GLP-1) and improves glycemic control in patients with type 2 diabetes. Although GLP-1 is known to stimulate insulin secretion, vildagliptin does not affect plasma insulin levels...... in diabetic patients, suggesting that more sophisticated measures are necessary to ascertain the influence of vildagliptin on beta-cell function. METHODS: This study examined the effects of 28-d treatment with vildagliptin (100 mg, twice daily; n = 9) vs. placebo (n = 11) on beta-cell function in diabetic...

  14. Risk factors and gene type for infections of MRSA in diabetic foot patients in Tianjin, China.

    Science.gov (United States)

    Feng, Shu-Hong; Chu, Yue-Jie; Wang, Peng-Hua; Jun, Xu; Min, Ding; Li, Xue-Mei

    2013-06-01

    The objective was to study risk factors and gene type of DF patients infected with MRSA. A total of 429 DF patients were recruited. The patients with S aureus infections were divided into MRSA and MSSA groups. MRSA were genotyped by SCCmec. pvl and lukE-lukD were detected. A total of 559 pathogens were isolated from them, with G+ bacteria firstly(59.0%), followed G- bacilli (37.7%) and true fungi (3.3%). The 3 most frequently isolated pathogens were S aureus (35.2%), S epidermidis (12.3%), and Pseudomonas aeruginosa (11.2%). SCCmec III MRSA and SCCmec IVa MRSA had the same antibacterial spectrum. mecA positive rate was 100%. lukE-lukD and pvl positive rates were 100% and 0%, respectively. 28 strains belonged to SCCmec III and the others belonged to SCCmec IVa. The G+ cocci were the main pathogens, S aureus and S epidermidis were predominant among them. Antibiotic usage in 6 months prior to hospitalization, long course of ulcer, osteomyelitis and hypoproteinemia are risk factors for MRSA. SCCmec IVa is high in proportion to MRSA isolates, suggesting that CA-MRSA has become major pathogen of DF infection. All the MRSA were harboring lukE-lukD, which has been reported to present poor leucotoxin compared to pvl, and may be a response to atypical local inflammatory reaction in DF infection.

  15. Salter-Harris type IV fracture of the proximal phalanx of the thumb with rotation of the epiphysis: Outcome 10 years following open reduction and K-wire fixation

    Directory of Open Access Journals (Sweden)

    Mohammad M. Al-Qattan

    2017-01-01

    Conclusion: We present a rare case of Salter-Harris type IV fracture of the proximal phalanx of the thumb with rotation of the epiphysis. Long term outcome was satisfactory but there was diaphyseal constriction leading to a narrower thumb.

  16. Comparative evaluation of few physical properties of epoxy resin, resin-modified gypsum and conventional type IV gypsum die materials: an in vitro study.

    Science.gov (United States)

    Gujjarlapudi, Manmohan Choudary; Reddy, S Varalakshmi; Madineni, Praveen Kumar; Ealla, Kranti Kiran Reddy; Nunna, Venkata Narayana; Manne, Sanjay Dutt

    2012-01-01

    To compare and evaluate few physical properties of epoxy resin, resin-modified gypsum and conventional type-IV gypsum die material. In the present study, dimensional accuracy, surface detail reproduction and transverse strength of three die materials like epoxy resin (Diemet-E), resin-modified gypsum (Synarock) and conventional type-IV gypsum (Ultrarock) are analyzed. For dimensional accuracy, master die (Bailey's die) is used and calibrations were made with digital microscope. For surface detail reproduction and transverse strength, rectangular stainless steel master die (Duke's die) was used and calibrations were made with Toolmaker's microscope and Instron universal testing machine respectively. One-way analysis of variance (ANOVA) was performed on the means and standard deviation for groups of each test. The results of the study showed statistically significant difference among these materials in dimensional accuracy, surface detail reproduction and transverse strength. Epoxy resin exhibited superiority in dimensional accuracy, surface detail reproduction and transverse strength and is nearest to the standards of accurate die material.

  17. Flagella but not type IV pili are involved in the initial adhesion of Pseudomonas aeruginosa PAO1 to hydrophobic or superhydrophobic surfaces.

    Science.gov (United States)

    Bruzaud, Jérôme; Tarrade, Jeanne; Coudreuse, Arnaud; Canette, Alexis; Herry, Jean-Marie; Taffin de Givenchy, Elisabeth; Darmanin, Thierry; Guittard, Frédéric; Guilbaud, Morgan; Bellon-Fontaine, Marie-Noëlle

    2015-07-01

    Over the last decades, surface biocontamination has become a major concern in food industries and medical environments where its outcomes could vary from financial losses to public health issues. Understanding adhesion mechanisms of involved microorganisms is essential to develop new strategies of prevention and control. Adhesion of Pseudomonas aeruginosa, a nosocomial pathogenic bacterium, relies on several bacterial features, among which are bacterial appendages such as flagella and type IV pili. Here, we examine the role of P. aeruginosa PAO1 flagella and type IV pili in the adhesion to abiotic surfaces with various hydrophobicities. Adhesion kinetics showed, that after 60min, flagella increased the adhesion of the strain to surfaces with high hydrophobicity while no effect was observed on hydrophilic surfaces. Flagella of adherent bacteria exhibited specific and conserved pattern on the surfaces that suggested a higher affinity of flagella for hydrophobic surfaces. Based on these results and on previous studies in the literature, we proposed a model of flagella-mediated adhesion onto hydrophobic surfaces where these appendages induce the first contact and promote the adhesion of the bacterial body. These findings suggest that anti-bioadhesive surface design should take into consideration the presence of bacterial appendages. Copyright © 2015 Elsevier B.V. All rights reserved.

  18. FOURCRACK advanced coal-fired power plant steels - avoidance of early weld failure by 'Type IV' cracking

    Energy Technology Data Exchange (ETDEWEB)

    Allen, D.J.; Harvey, B.; Brett, S.J. [E.ON UK plc., Nottingham (United Kingdom)

    2005-12-15

    Project 'FOURCRACK' was undertaken to improve understanding of the problem, assist materials selection, and clarify the effective limits on plant design conditions and temperatures. The main project was based on cross-weld creep rupture testing. Tests were carried out to assess the high temperature performance of welds in seven different casts of leading advanced high alloy ferritic steels, E911, P92 and P122, supplied as forging, casting, plate, and pipe components, and in a weak cast of Grade 91 steel which was welded following different heat treatments to compare the performance of as received, renormalised and tempered and preaged parent material conditions. A novel form of test matrix was devised to investigate variations in both stress and temperature and compare the performance of a large number of different weldment over a range of different test conditions. To generate better long term data on Type IV cracking in the heat-affected zone, the main test programme was followed by additional testing using a special waited specimen geometry. The test data included many short term failures in the weld metal. However, there was a clear trend toward HAZ failure at lower temperature and stresses, and it could be concluded that HAZ Type IV failure is the main long term risk under plant operating conditions. 15 refs., 29 figs., 26 abs., 5 apps.

  19. A Posteriorly Displaced Distal Metaphyseal Clavicular Fracture (Type IV AC Joint Dislocation-Like in Children: A Case Report and Literature Review Study

    Directory of Open Access Journals (Sweden)

    Ahmed Kotb

    2016-01-01

    Full Text Available Fractures of the lateral end of the clavicle are common in pediatric patients; most of these fractures occur at the physeal level representing Salter Harris injuries. The vast majority of fractures of the lateral end of the clavicle are managed nonoperatively. In this report, we describe a unique type of fracture of the distal end of the clavicle in the pediatric patients in which the fracture occurs in the metaphyseal lateral clavicle with the proximal edge of the fracture displaced posteriorly through the trapezius muscle causing obvious deformity. It is similar in pathology to type IV AC joint dislocation. In this study we report this injury in eleven-year-old boy. Literature review showed that similar injuries were described before three times (two of them in pediatric patients. Due to the significant clinical deformity of this category with entrapment of the bone through the trapezius muscle, reduction (open or closed of the fracture is the recommended treatment.

  20. The Sloan Lens ACS survey. IV. The mass density profile of early-type galaxies out to 100 effective radii

    NARCIS (Netherlands)

    Gavazzi, Raphael; Treu, Tommaso; Rhodes, Jason D.; Koopmans, Leon V. E.; Bolton, Adam S.; Burles, Scott; Massey, Richard J.; Moustakas, Leonidas A.

    2007-01-01

    We present a weak-lensing analysis of 22 early-type (strong) lens galaxies, based on deep HST images obtained as part of the Sloan Lens ACS Survey. Using advanced techniques to control systematic uncertainties, we show that weak-lensing signal is detected out to similar to 300 h(-1) kpc (at the mean

  1. Glucagon-like peptide 1 and inhibitors of dipeptidyl peptidase IV in the treatment of type 2 diabetes mellitus

    DEFF Research Database (Denmark)

    Holst, Jens Juul; Deacon, Carolyn F

    2004-01-01

    Proof-of-concept for the efficacy of a glucagon-like peptide 1 (GLP-1)-based therapy of patients with type 2 diabetes was provided in 2002 by means of prolonged continuous subcutaneous infusion of native GLP-1. Since then, several long-acting analogues of GLP-1, as well as inhibitors of dipeptidyl...

  2. Serum Levels of Soluble CD26/Dipeptidyl Peptidase-IV in Type 2 Diabetes Mellitus and Its Association with Metabolic Syndrome and Therapy with Antidiabetic Agents in Malaysian Subjects

    Science.gov (United States)

    Ahmed, Radwan H.; Huri, Hasniza Zaman; Al-Hamodi, Zaid; Salem, Sameer D.; Muniandy, Sekaran

    2015-01-01

    Background A soluble form of CD26/dipeptidyl peptidase-IV (sCD26/DPP-IV) induces DPP-IV enzymatic activity that degrades incretin. We investigated fasting serum levels of sCD26/DPP-IV and active glucagon-like peptide-1 (GLP-1) in Malaysian patients with type 2 diabetes mellitus (T2DM) with and without metabolic syndrome (MetS), as well as the associations between sCD26/DPP-IV levels, MetS, and antidiabetic therapy. Methods We assessed sCD26/DPP-IV levels, active GLP-1 levels, body mass index (BMI), glucose, insulin, A1c, glucose homeostasis indices, and lipid profiles in 549 Malaysian subjects (including 257 T2DM patients with MetS, 57 T2DM patients without MetS, 71 non-diabetics with MetS, and 164 control subjects without diabetes or metabolic syndrome). Results Fasting serum levels of sCD26/DPP-IV were significantly higher in T2DM patients with and without MetS than in normal subjects. Likewise, sCD26/DPP-IV levels were significantly higher in patients with T2DM and MetS than in non-diabetic patients with MetS. However, active GLP-1 levels were significantly lower in T2DM patients both with and without MetS than in normal subjects. In T2DM subjects, sCD26/DPP-IV levels were associated with significantly higher A1c levels, but were significantly lower in patients using monotherapy with metformin. In addition, no significant differences in sCD26/DPP-IV levels were found between diabetic subjects with and without MetS. Furthermore, sCD26/DPP-IV levels were negatively correlated with active GLP-1 levels in T2DM patients both with and without MetS. In normal subjects, sCD26/DPP-IV levels were associated with increased BMI, cholesterol, and LDL-cholesterol (LDL-c) levels. Conclusion Serum sCD26/DPP-IV levels increased in T2DM subjects with and without MetS. Active GLP-1 levels decreased in T2DM patients both with and without MetS. In addition, sCD26/DPP-IV levels were associated with Alc levels and negatively correlated with active GLP-1 levels. Moreover, metformin

  3. The Helicobacter pylori type IV secretion system promotes IL-8 synthesis in a model of pediatric airway epithelium via p38 MAP kinase.

    Directory of Open Access Journals (Sweden)

    Myra G Dela Pena-Ponce

    Full Text Available Epidemiologic studies have reported an inverse relationship between childhood Helicobacter pylori infection and development of allergic asthma. Because lung epithelium plays an important role in allergic asthma pathogenesis, we hypothesized that H. pylori may directly influence airway epithelial cell innate immune function, particularly in early childhood. To test our hypothesis, we established an in vitro H. pylori infection model using primary tracheobronchial epithelial cell cultures derived from infant, juvenile and adult rhesus monkeys. Airway epithelial cell cultures were infected with wild-type or cag pathogenicity island mutant H. pylori strains, followed by evaluation of IL-8 and IL-6 protein synthesis. We found that H. pylori primarily increased IL-8 synthesis in a MOI and age-dependent fashion, with a greater than 4-fold induction in infant versus adult cultures. H. pylori-induced IL-8 synthesis in infant and juvenile cultures was significantly reduced by cag pathogenicity island mutants, indicating a requirement for the type IV secretion system. Although peptidoglycan recognition of nucleotide binding oligomerization domain-containing protein 1 (NOD1 and NF-kappaB have been implicated as key cytokine signaling molecules for H. pylori infection in gastric epithelium, NOD1 (ML130 or NF-kappaB (JSH-23 inhibitors minimally affected IL-8 synthesis in airway epithelial cell cultures following H. pylori infection. In contrast, inhibition of the p38 MAP kinase pathway (SB203580 resulted in almost complete suppression of H. pylori-induced IL-8 synthesis. Collectively, these results indicate that H. pylori can preferentially elicit IL-8 synthesis in a model of pediatric airway epithelium using the type IV secretion system via p38 MAP kinase.

  4. SDSS-IV MaNGA: Spatially resolved star formation histories in galaxies as a function of galaxy mass and type

    Science.gov (United States)

    Goddard, D.; Thomas, D.; Maraston, C.; Westfall, K.; Etherington, J.; Riffel, R.; Mallmann, N. D.; Zheng, Z.; Argudo-Fernández, M.; Lian, J.; Bershady, M.; Bundy, K.; Drory, N.; Law, D.; Yan, R.; Wake, D.; Weijmans, A.; Bizyaev, D.; Brownstein, J.; Lane, R. R.; Maiolino, R.; Masters, K.; Merrifield, M.; Nitschelm, C.; Pan, K.; Roman-Lopes, A.; Storchi-Bergmann, T.; Schneider, D. P.

    2017-04-01

    We study the internal gradients of stellar population properties within 1.5 Re for a representative sample of 721 galaxies, with stellar masses ranging between 109 M⊙ and 1011.5 M⊙ from the SDSS-IV MaNGA Integral-Field-Unit survey. Through the use of our full spectral fitting code firefly, we derive light- and mass-weighted stellar population properties and their radial gradients, as well as full star formation and metal enrichment histories. We also quantify the impact that different stellar population models and full spectral fitting routines have on the derived stellar population properties and the radial gradient measurements. In our analysis, we find that age gradients tend to be shallow for both early-type and late-type galaxies. Mass-weighted age gradients of early-types arepositive (˜0.09 dex/Re) pointing to 'outside-in' progression of star formation, while late-type galaxies have negative light-weighted age gradients (˜-0.11 dex/Re), suggesting an 'inside-out' formation of discs. We detect negative metallicity gradients in both early- and late-type galaxies, but these are significantly steeper in late-types, suggesting that the radial dependence of chemical enrichment processes and the effect of gas inflow and metal transport are far more pronounced in discs. Metallicity gradients of both morphological classes correlate with galaxy mass, with negative metallicity gradients becoming steeper with increasing galaxy mass. The correlation with mass is stronger for late-type galaxies, with a slope of d(∇[Z/H])/d(log M) ˜ -0.2 ± 0.05 , compared to d(∇[Z/H])/d(log M) ˜ -0.05 ± 0.05 for early-types. This result suggests that the merger history plays a relatively small role in shaping metallicity gradients of galaxies.

  5. Distribution of collagen types I, III, and IV in gastric tissue of marmosets (Callithrix spp., Callitrichidae: Primates)

    OpenAIRE

    Mello, Marcela F.V. de; Pissinatti, Alcides; Ferreira, Ana M.R.

    2010-01-01

    Extracellular matrix (ECM) components such as fibrillar collagens play a fundamental role in wound repair and have also been studied in association with the gastric ulcer healing process in gastroenterology. Nevertheless, there have been no studies in the literature to date regarding the description and characterization of ECM components, neither in normal nor in injured gastric tissue of primate species. The objective of this study was to investigate the expression of gastric collagen types ...

  6. Analysis of the type IV fimbrial-subunit gene fimA of Xanthomonas hyacinthi: application in PCR-mediated detection of yellow disease in Hyacinths.

    Science.gov (United States)

    van Doorn, J; Hollinger, T C; Oudega, B

    2001-02-01

    A sensitive and specific detection method was developed for Xanthomonas hyacinthi; this method was based on amplification of a subsequence of the type IV fimbrial-subunit gene fimA from strain S148. The fimA gene was amplified by PCR with degenerate DNA primers designed by using the N-terminal and C-terminal amino acid sequences of trypsin fragments of FimA. The nucleotide sequence of fimA was determined and compared with the nucleotide sequences coding for the fimbrial subunits in other type IV fimbria-producing bacteria, such as Xanthomonas campestris pv. vesicatoria, Neisseria gonorrhoeae, and Moraxella bovis. In a PCR internal primers JAAN and JARA, designed by using the nucleotide sequences of the variable central and C-terminal region of fimA, amplified a 226-bp DNA fragment in all X. hyacinthi isolates. This PCR was shown to be pathovar specific, as assessed by testing 71 Xanthomonas pathovars and bacterial isolates belonging to other genera, such as Erwinia and Pseudomonas. Southern hybridization experiments performed with the labelled 226-bp DNA amplicon as a probe suggested that there is only one structural type IV fimbrial-gene cluster in X. hyacinthi. Only two Xanthomonas translucens pathovars cross-reacted weakly in PCR. Primers amplifying a subsequence of the fimA gene of X. campestris pv. vesicatoria (T. Ojanen-Reuhs, N. Kalkkinen, B. Westerlund-Wikström, J. van Doorn, K. Haahtela, E.-L. Nurmiaho-Lassila, K. Wengelink, U. Bonas, and T. K. Korhonen, J. Bacteriol. 179: 1280-1290, 1997) were shown to be pathovar specific, indicating that the fimbrial-subunit sequences are more generally applicable in xanthomonads for detection purposes. Under laboratory conditions, approximately 1,000 CFU of X. hyacinthi per ml could be detected. In inoculated leaves of hyacinths the threshold was 5,000 CFU/ml. The results indicated that infected hyacinths with early symptoms could be successfully screened for X. hyacinthi with PCR.

  7. Structures of acetylated oleanane-type triterpene saponins, rarasaponins IV, V, and VI, and anti-hyperlipidemic constituents from the pericarps of Sapindus rarak.

    Science.gov (United States)

    Asao, Yasunobu; Morikawa, Toshio; Xie, Yuanyuan; Okamoto, Masaki; Hamao, Makoto; Matsuda, Hisashi; Muraoka, Osamu; Yuan, Dan; Yoshikawa, Masayuki

    2009-02-01

    The methanolic extract and its saponin fraction (methanol-eluted fraction) of the pericarps of Sapindus rarak DC. were found to suppress plasma triglyceride elevation in olive oil-treated mice. From the active fraction, three new acylated oleanane-type triterpene saponins, rarasaponins IV (1), V (2), and VI (3), were isolated. The structures of 1-3 were elucidated on the basis of chemical and spectroscopic evidence. The principle saponin constituents, hederagenin 3-O-alpha-L-arabinopyranosyl-(1-->3)-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranoside (4) and hederagenin 3-O-(3,4-di-O-acetyl-alpha-L-arabinopyranosyl)-(1-->3)-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranoside (5), showed inhibitory effects on plasma triglyceride elevation at a dose of 200 mg/kg, per os.

  8. Osteogenesis imperfecta Type IV: a newly identified variant at position c.560 (G > T; p.Gly187Val) in the COL1A2 gene.

    Science.gov (United States)

    Usta, Akin; Karademir, Dilay; Sen, Eylem; Yazici, Selcuk; Adali, Ertan; Erdem, Erkan; Karacan, Meric

    2017-01-01

    Osteogenesis imperfecta is a clinically heterogenous disease caused by defective collagen syntesis associated with a mutation in the COL1A1 or COL1A2 genes. In this report, we present a case of osteogenesis imperfecta (OI) type IV, seen in a female fetus with incurved femurs at 18 weeks of gestation. Molecular analysis of the newborn revealed a novel mutation at position c.560 (c.560 G > T) of the exon 12 in the COL1A2 gene; which lead to the glycine modification with valine (p.Gly187Val) at codon 187. The pregnancy follow-up was uneventful. After delivery, the newborn underwent biphosponat therapy and no fracture was detected until 1 year old.

  9. Low-energy helium-neon laser induces melanocyte proliferation via interaction with type IV collagen: visible light as a therapeutic option for vitiligo.

    Science.gov (United States)

    Lan, C-C E; Wu, C-S; Chiou, M-H; Chiang, T-Y; Yu, H-S

    2009-08-01

    The treatment of vitiligo remains a challenge for clinical dermatologists. We have previously shown that the helium-neon laser (He-Ne laser, 632.8 nm) is a therapeutic option for treatment of this depigmentary disorder. Addressing the intricate interactions between melanocytes, the most important cellular component in the repigmentation scheme of vitiligo, and their innate extracellular matrix collagen type IV, the current study aimed to elucidate the effects of the He-Ne laser on melanocytes. Cultured melanocytes were irradiated with the He-Ne laser. Relevant biological parameters including cell attachment, locomotion and growth were evaluated. In addition, the potentially involved molecular pathways were also determined. Our results show that in addition to suppressing mobility but increasing attachment to type IV collagen, the He-Ne laser stimulates melanocyte proliferation through enhanced alpha2beta1 integrin expression. The expression of phosphorylated cyclic-AMP response element binding protein (CREB), an important regulator of melanocyte growth, was also upregulated by He-Ne laser treatment. Using a specific mitochondrial uncoupling agent [carbonyl cyanide m-chlorophenyl-hydrazone (CCCP)], the proliferative effect of the He-Ne laser on melanocytes was abolished and suppression of melanocyte growth was noted. In summary, we have demonstrated that the He-Ne laser imparts a growth stimulatory effect on functional melanocytes via mitochondria-related pathways and proposed that other minor pathways including DNA damage may also be inflicted by laser treatment on irradiated cells. More importantly, we have completed the repigmentation scheme of vitiligo brought about by He-Ne laser light in vitro and provided a solid theoretical basis regarding how the He-Ne laser induces recovery of vitiligo in vivo.

  10. Experimental Investigation of Secondary Flow Structures Downstream of a Model Type IV Stent Failure in a 180° Curved Artery Test Section.

    Science.gov (United States)

    Bulusu, Kartik V; Plesniak, Michael W

    2016-07-19

    The arterial network in the human vasculature comprises of ubiquitously present blood vessels with complex geometries (branches, curvatures and tortuosity). Secondary flow structures are vortical flow patterns that occur in curved arteries due to the combined action of centrifugal forces, adverse pressure gradients and inflow characteristics. Such flow morphologies are greatly affected by pulsatility and multiple harmonics of physiological inflow conditions and vary greatly in size-strength-shape characteristics compared to non-physiological (steady and oscillatory) flows (1 - 7). Secondary flow structures may ultimately influence the wall shear stress and exposure time of blood-borne particles toward progression of atherosclerosis, restenosis, sensitization of platelets and thrombosis (4 - 6, 8 - 13). Therefore, the ability to detect and characterize these structures under laboratory-controlled conditions is precursor to further clinical investigations. A common surgical treatment to atherosclerosis is stent implantation, to open up stenosed arteries for unobstructed blood flow. But the concomitant flow perturbations due to stent installations result in multi-scale secondary flow morphologies (4 - 6). Progressively higher order complexities such as asymmetry and loss in coherence can be induced by ensuing stent failures vis-à-vis those under unperturbed flows (5). These stent failures have been classified as "Types I-to-IV" based on failure considerations and clinical severity (14). This study presents a protocol for the experimental investigation of the complex secondary flow structures due to complete transverse stent fracture and linear displacement of fractured parts ("Type IV") in a curved artery model. The experimental method involves the implementation of particle image velocimetry (2C-2D PIV) techniques with an archetypal carotid artery inflow waveform, a refractive index matched blood-analog working fluid for phase-averaged measurements (15 - 18

  11. Inulin-type fructan degradation capacity of Clostridium cluster IV and XIVa butyrate-producing colon bacteria and their associated metabolic outcomes.

    Science.gov (United States)

    Moens, F; De Vuyst, L

    2017-05-30

    Four selected butyrate-producing colon bacterial strains belonging to Clostridium cluster IV (Butyricicoccus pullicaecorum DSM 23266 T and Faecalibacterium prausnitzii DSM 17677 T ) and XIVa (Eubacterium hallii DSM 17630 and Eubacterium rectale CIP 105953 T ) were studied as to their capacity to degrade inulin-type fructans and concomitant metabolite production. Cultivation of these strains was performed in bottles and fermentors containing a modified medium for colon bacteria, including acetate, supplemented with either fructose, oligofructose, or inulin as the sole energy source. Inulin-type fructan degradation was not a general characteristic among these strains. B. pullicaecorum DSM 23266 T and E. hallii DSM 17630 could only ferment fructose and did not degrade oligofructose or inulin. E. rectale CIP 105953 T and F. prausnitzii DSM 17677 T fermented fructose and could degrade both oligofructose and inulin. All chain length fractions of oligofructose were degraded simultaneously (both strains) and both long and short chain length fractions of inulin were degraded either simultaneously (E. rectale CIP 105953 T ) or consecutively (F. prausnitzii DSM 17677 T ), indicating an extracellular polymer degradation mechanism. B. pullicaecorum DSM 23266 T and E. hallii DSM 17630 produced high concentrations of butyrate, CO 2 , and H 2 from fructose. E. rectale CIP 105953 T produced lactate, butyrate, CO 2 , and H 2 , from fructose, oligofructose, and inulin, whereas F. prausnitzii DSM 17677 T produced butyrate, formate, CO 2 , and traces of lactate from fructose, oligofructose, and inulin. Based on carbon recovery and theoretical metabolite production calculations, an adapted stoichiometrically balanced metabolic pathway for butyrate, formate, lactate, CO 2 , and H 2 production by members of both Clostridium cluster IV and XIVa butyrate-producing bacteria was constructed.

  12. SDSS-IV MaNGA: Variation of the Stellar Initial Mass Function in Spiral and Early-type Galaxies

    Science.gov (United States)

    Li, Hongyu; Ge, Junqiang; Mao, Shude; Cappellari, Michele; Long, R. J.; Li, Ran; Emsellem, Eric; Dutton, Aaron A.; Li, Cheng; Bundy, Kevin; Thomas, Daniel; Drory, Niv; Lopes, Alexandre Roman

    2017-04-01

    We perform Jeans anisotropic modeling (JAM) on elliptical and spiral galaxies from the MaNGA DR13 sample. By comparing the stellar mass-to-light ratios estimated from stellar population synthesis and from JAM, we find a systematic variation of the initial mass function (IMF) similar to that in the earlier {{ATLAS}}3{{D}} results. Early-type galaxies (elliptical and lenticular) with lower velocity dispersions within one effective radius are consistent with a Chabrier-like IMF, while galaxies with higher velocity dispersions are consistent with a more bottom-heavy IMF such as the Salpeter IMF. Spiral galaxies have similar systematic IMF variations, but with slightly different slopes and larger scatters, due to the uncertainties caused by the higher gas fractions and extinctions for these galaxies. Furthermore, we examine the effects of stellar mass-to-light ratio gradients on our JAM modeling, and we find that the trends become stronger after considering the gradients.

  13. SDSS-IV MaNGA: Uncovering the Angular Momentum Content of Central and Satellite Early-type Galaxies

    Science.gov (United States)

    Greene, J. E.; Leauthaud, A.; Emsellem, E.; Ge, J.; Aragón-Salamanca, A.; Greco, J.; Lin, Y.-T.; Mao, S.; Masters, K.; Merrifield, M.; More, S.; Okabe, N.; Schneider, D. P.; Thomas, D.; Wake, D. A.; Pan, K.; Bizyaev, D.; Oravetz, D.; Simmons, A.; Yan, R.; van den Bosch, F.

    2018-01-01

    We study 379 central and 159 satellite early-type galaxies with two-dimensional kinematics from the integral-field survey Mapping Nearby Galaxies at APO (MaNGA) to determine how their angular momentum content depends on stellar and halo mass. Using the Yang et al. group catalog, we identify central and satellite galaxies in groups with halo masses in the range {10}12.5 {h}-1 {M}ȯ {10}11 {h}-2 {M}ȯ tend to have very little rotation, while nearly all galaxies at lower mass show some net rotation. The ∼30% of high-mass galaxies that have significant rotation do not stand out in other galaxy properties, except for a higher incidence of ionized gas emission. Our data are consistent with recent simulation results suggesting that major merging and gas accretion have more impact on the rotational support of lower-mass galaxies. When carefully matching the stellar mass distributions, we find no residual differences in angular momentum content between satellite and central galaxies at the 20% level. Similarly, at fixed mass, galaxies have consistent rotation properties across a wide range of halo mass. However, we find that errors in classification of central and satellite galaxies with group finders systematically lower differences between satellite and central galaxies at a level that is comparable to current measurement uncertainties. To improve constraints, the impact of group-finding methods will have to be forward-modeled via mock catalogs.

  14. A dual injury of the shoulder: acromioclavicular joint dislocation (type IV) coupled with ipsilateral mid-shaft clavicle fracture.

    Science.gov (United States)

    Madi, Sandesh; Pandey, Vivek; Khanna, Vikrant; Acharya, Kiran

    2015-11-23

    A direct blow to the shoulder, as may be sustained in a road traffic accident (RTA), can result in various combinations of fracture dislocations in the shoulder joint complex. Among these, a rare variety is an acromioclavicular joint (ACJ) dislocation coupled with ipsilateral mid-shaft clavicle fracture. Diverse treatment options have been described in the literature, ranging from non-operative and operative, to hybrid management. Treatment for this complex injury is predominantly dictated by the type of dislocation and displacement of the clavicle fracture, as well as age and demand of the patient. Acute high grades of ACJ dislocation require restoration of the coracoclavicular relationship (in place of torn coracoclavicular (CC) ligament) by some form of internal fixation, thereby maintaining the ACJ reduction. An arthroscopic reinstatement of the coracoclavicular relationship using a dog bone button and fibre tape implant for this composite injury pattern has not been previously described. Furthermore, a comprehensive review of the literature associated with this injury pattern is briefly described. 2015 BMJ Publishing Group Ltd.

  15. Repeated shock stress facilitates basolateral amygdala synaptic plasticity through decreased cAMP-specific phosphodiesterase type IV (PDE4) expression.

    Science.gov (United States)

    Ryan, Steve; Li, Chenchen; Menigoz, Aurélie; Hazra, Rimi; Dabrowska, Joanna; Ehrlich, David; Gordon, Katelyn; Rainnie, Donald G

    2017-12-04

    Previous studies have shown that exposure to stressful events can enhance fear memory and anxiety-like behavior as well as increase synaptic plasticity in the rat basolateral amygdala (BLA). We have evidence that repeated unpredictable shock stress (USS) elicits a long-lasting increase in anxiety-like behavior in rats, but the cellular mechanisms mediating this response remain unclear. Evidence from recent morphological studies suggests that alterations in the dendritic arbor or spine density of BLA principal neurons may underlie stress-induced anxiety behavior. Recently, we have shown that the induction of long-term potentiation (LTP) in BLA principal neurons is dependent on activation of postsynaptic D1 dopamine receptors and the subsequent activation of the cyclic adenosine 5'-monophosphate (cAMP)-protein kinase A (PKA) signaling cascade. Here, we have used in vitro whole-cell patch-clamp recording from BLA principal neurons to investigate the long-term consequences of USS on their morphological properties and synaptic plasticity. We provided evidence that the enhanced anxiety-like behavior in response to USS was not associated with any significant change in the morphological properties of BLA principal neurons, but was associated with a changed frequency dependence of synaptic plasticity, lowered LTP induction threshold, and reduced expression of phosphodiesterase type 4 enzymes (PDE4s). Furthermore, pharmacological inhibition of PDE4 activity with rolipram mimics the effects of chronic stress on LTP induction threshold and baseline startle. Our results provide the first evidence that stress both enhances anxiety-like behavior and facilitates synaptic plasticity in the amygdala through a common mechanism of PDE4-mediated disinhibition of cAMP-PKA signaling.

  16. Parental somatic and germ-line mosaicism for a multiexon deletion with unusual endpoints in a type III collagen (COL3Al) allele produces ehlers-danlos syndrome type IV in the heterozygous offspring

    Energy Technology Data Exchange (ETDEWEB)

    McGookey Milewicz, D.; Witz, A.M.; Byers, P.H. (Univ of Washington, Seattle (United States)); Smith, A.C.M.; Manchester, D.K.; Waldstein, G. (Children' s Hospital, Denver, CO (United States))

    1993-07-01

    Ehlers-Danlos syndrome (EDS) type IV is a dominantly inherited disorder that results from mutation in the type III collagen gene (COL3A1). The authors studied the structure of the COL3A1 gene of an individual with EDS type IV and that of her phenotypically normal parents. The proband was heterozygous for a 2-kb deletion in COL3A1, while her father was mosaic for the same deletion in somatic and germ cells. In fibroblasts from the father, approximately two-fifths of the COL3A1 alleles carried the deletion, but only 10% of the COL3A1 alleles in white blood cells were of the mutant species. The deletion in the mutant allele extended from intron 7 into intron 11. There was a 12-bp direct repeat in intron 7 and intron 11, the latter about 60 bp 5' to the junction. At the breakpoint there was a duplication of 10 bp from intron 11 separated by an insertion of 4 bp contained within the duplicated sequence. The father was mosaic for the deletion so that the gene rearrangement occurred during his early embryonic development prior to lineage allocation. These findings suggest that at least some of the deletions seen in human genes may occur during replication, rather than as a consequence of meiotic crossing-over, and that they thus have a risk for recurrence when observed de novo. 71 refs., 4 figs., 2 tabs.

  17. Phase IV, Open-Label, Safety Study Evaluating the Use of Dexmedetomidine in Pediatric Patients Undergoing Procedure-Type Sedation

    Directory of Open Access Journals (Sweden)

    Edmund H. Jooste

    2017-08-01

    dropped by 10% in two patients, but resolved without need for manual ventilation. All other reported AEs were consistent with the known safety profile of dexmedetomidine. Two of the 78 patients in the efficacy-evaluable population met all sedation efficacy criteria. Dexmedetomidine was well-tolerated in pediatric patients undergoing procedure-type sedation.

  18. Studying Coxiella burnetii Type IV Substrates in the Yeast Saccharomyces cerevisiae: Focus on Subcellular Localization and Protein Aggregation.

    Science.gov (United States)

    Rodríguez-Escudero, María; Cid, Víctor J; Molina, María; Schulze-Luehrmann, Jan; Lührmann, Anja; Rodríguez-Escudero, Isabel

    2016-01-01

    Coxiella burnetii is a Gram-negative obligate parasitic bacterium that causes the disease Q-fever in humans. To establish its intracellular niche, it utilizes the Icm/Dot type IVB secretion system (T4BSS) to inject protein effectors into the host cell cytoplasm. The host targets of most cognate and candidate T4BSS-translocated effectors remain obscure. We used the yeast Saccharomyces cerevisiae as a model to express and study six C. burnetii effectors, namely AnkA, AnkB, AnkF, CBU0077, CaeA and CaeB, in search for clues about their role in C. burnetii virulence. When ectopically expressed in HeLa cells, these effectors displayed distinct subcellular localizations. Accordingly, GFP fusions of these proteins produced in yeast also decorated distinct compartments, and most of them altered cell growth. CaeA was ubiquitinated both in yeast and mammalian cells and, in S. cerevisiae, accumulated at juxtanuclear quality-control compartments (JUNQs) and insoluble protein deposits (IPODs), characteristic of aggregative or misfolded proteins. AnkA, which was not ubiquitinated, accumulated exclusively at the IPOD. CaeA, but not AnkA or the other effectors, caused oxidative damage in yeast. We discuss that CaeA and AnkA behavior in yeast may rather reflect misfolding than recognition of conserved targets in the heterologous system. In contrast, CBU0077 accumulated at vacuolar membranes and abnormal ER extensions, suggesting that it interferes with vesicular traffic, whereas AnkB associated with the yeast nucleolus. Both effectors shared common localization features in HeLa and yeast cells. Our results support the idea that C. burnetii T4BSS effectors manipulate multiple host cell targets, which can be conserved in higher and lower eukaryotic cells. However, the behavior of CaeA and AnkA prompt us to conclude that heterologous protein aggregation and proteostatic stress can be a limitation to be considered when using the yeast model to assess the function of bacterial effectors.

  19. A novel COL4A1 frameshift mutation in familial kidney disease: the importance of the C-terminal NC1 domain of type IV collagen

    Science.gov (United States)

    Gale, Daniel P.; Oygar, D. Deren; Lin, Fujun; Oygar, P. Derin; Khan, Nadia; Connor, Thomas M.F.; Lapsley, Marta; Maxwell, Patrick H.; Neild, Guy H.

    2016-01-01

    Background Hereditary microscopic haematuria often segregates with mutations of COL4A3, COL4A4 or COL4A5 but in half of families a gene is not identified. We investigated a Cypriot family with autosomal dominant microscopic haematuria with renal failure and kidney cysts. Methods We used genome-wide linkage analysis, whole exome sequencing and cosegregation analyses. Results We identified a novel frameshift mutation, c.4611_4612insG:p.T1537fs, in exon 49 of COL4A1. This mutation predicts truncation of the protein with disruption of the C-terminal part of the NC1 domain. We confirmed its presence in 20 family members, 17 with confirmed haematuria, 5 of whom also had stage 4 or 5 chronic kidney disease. Eleven family members exhibited kidney cysts (55% of those with the mutation), but muscle cramps or cerebral aneurysms were not observed and serum creatine kinase was normal in all individuals tested. Conclusions Missense mutations of COL4A1 that encode the CB3 [IV] segment of the triple helical domain (exons 24 and 25) are associated with HANAC syndrome (hereditary angiopathy, nephropathy, aneurysms and cramps). Missense mutations of COL4A1 that disrupt the NC1 domain are associated with antenatal cerebral haemorrhage and porencephaly, but not kidney disease. Our findings extend the spectrum of COL4A1 mutations linked with renal disease and demonstrate that the highly conserved C-terminal part of the NC1 domain of the α1 chain of type IV collagen is important in the integrity of glomerular basement membrane in humans. PMID:27190376

  20. Chimeric Coupling Proteins Mediate Transfer of Heterologous Type IV Effectors through the Escherichia coli pKM101-Encoded Conjugation Machine.

    Science.gov (United States)

    Whitaker, Neal; Berry, Trista M; Rosenthal, Nathan; Gordon, Jay E; Gonzalez-Rivera, Christian; Sheehan, Kathy B; Truchan, Hilary K; VieBrock, Lauren; Newton, Irene L G; Carlyon, Jason A; Christie, Peter J

    2016-10-01

    Bacterial type IV secretion systems (T4SSs) are composed of two major subfamilies, conjugation machines dedicated to DNA transfer and effector translocators for protein transfer. We show here that the Escherichia coli pKM101-encoded conjugation system, coupled with chimeric substrate receptors, can be repurposed for transfer of heterologous effector proteins. The chimeric receptors were composed of the N-terminal transmembrane domain of pKM101-encoded TraJ fused to soluble domains of VirD4 homologs functioning in Agrobacterium tumefaciens, Anaplasma phagocytophilum, or Wolbachia pipientis A chimeric receptor assembled from A. tumefaciens VirD4 (VirD4At) mediated transfer of a MOBQ plasmid (pML122) and A. tumefaciens effector proteins (VirE2, VirE3, and VirF) through the pKM101 transfer channel. Equivalent chimeric receptors assembled from the rickettsial VirD4 homologs similarly supported the transfer of known or candidate effectors from rickettsial species. These findings establish a proof of principle for use of the dedicated pKM101 conjugation channel, coupled with chimeric substrate receptors, to screen for translocation competency of protein effectors from recalcitrant species. Many T4SS receptors carry sequence-variable C-terminal domains (CTDs) with unknown function. While VirD4At and the TraJ/VirD4At chimera with their CTDs deleted supported pML122 transfer at wild-type levels, ΔCTD variants supported transfer of protein substrates at strongly diminished or elevated levels. We were unable to detect binding of VirD4At's CTD to the VirE2 effector, although other VirD4At domains bound this substrate in vitro We propose that CTDs evolved to govern the dynamics of substrate presentation to the T4SS either through transient substrate contacts or by controlling substrate access to other receptor domains. Bacterial type IV secretion systems (T4SSs) display striking versatility in their capacity to translocate DNA and protein substrates to prokaryotic and

  1. Gene Flow Across Genus Barriers - Conjugation of Dinoroseobacter shibae's 191-kb Killer Plasmid into Phaeobacter inhibens and AHL-mediated Expression of Type IV Secretion Systems.

    Science.gov (United States)

    Patzelt, Diana; Michael, Victoria; Päuker, Orsola; Ebert, Matthias; Tielen, Petra; Jahn, Dieter; Tomasch, Jürgen; Petersen, Jörn; Wagner-Döbler, Irene

    2016-01-01

    Rhodobacteraceae harbor a conspicuous wealth of extrachromosomal replicons (ECRs) and therefore the exchange of genetic material via horizontal transfer has been supposed to be a major evolutionary driving force. Many plasmids in this group encode type IV secretion systems (T4SS) that are expected to mediate transfer of proteins and/or DNA into host cells, but no experimental evidence of either has yet been provided. Dinoroseobacter shibae, a species of the Roseobacter group within the Rhodobacteraceae family, contains five ECRs that are crucial for anaerobic growth, survival under starvation and the pathogenicity of this model organism. Here we tagged two syntenous but compatible RepABC-type plasmids of 191 and 126-kb size, each encoding a T4SS, with antibiotic resistance genes and demonstrated their conjugational transfer into a distantly related Roseobacter species, namely Phaeobacter inhibens. Pulsed field gel electrophoresis showed transfer of those replicons into the recipient both individually but also together documenting the efficiency of conjugation. We then studied the influence of externally added quorum sensing (QS) signals on the expression of the T4SS located on the sister plasmids. A QS deficient D. shibae null mutant (ΔluxI1 ) lacking synthesis of N-acyl-homoserine lactones (AHLs) was cultivated with a wide spectrum of chemically diverse long-chain AHLs. All AHLs with lengths of the acid side-chain ≥14 reverted the ΔluxI1 phenotype to wild-type. Expression of the T4SS was induced up to log2 ∼3fold above wild-type level. We hypothesize that conjugation in roseobacters is QS-controlled and that the QS system may detect a wide array of long-chain AHLs at the cell surface.

  2. Analysis of SLC40A1 gene at the mRNA level reveals rapidly the causative mutations in patients with hereditary hemochromatosis type IV.

    Science.gov (United States)

    Speletas, Matthaios; Kioumi, Anna; Loules, Gedeon; Hytiroglou, Prodromos; Tsitouridis, John; Christakis, John; Germenis, Anastasios E

    2008-01-01

    Mutations in the SLC40A1 gene result in a dominant genetic disorder [ferroportin disease; hereditary hemochromatosis type (HH) IV], characterized by iron overload with two different clinical manifestations, normal transferrin saturation with macrophage iron accumulation (the most prevalent type) or high transferrin saturation with hepatocyte iron accumulation (classical hemochromatosis phenotype). In previous studies, the mutational analysis of SLC40A1 gene has been performed at the genomic DNA level by PCR amplification and direct sequencing of all coding regions and flanking intron-exon boundaries (usually in 9 PCR reactions). In this study, we analyzed the SLC40A1 gene at the mRNA level, in two RT-PCR reactions, followed by direct sequencing and/or NIRCA (non-isotopic RNase cleavage assay). This protocol turned out to be rapid, sensitive and reliable, facilitating the detection of the SLC40A1 gene mutations in two patients with hyperferritinemia, normal transferrin saturation and iron accumulation predominantly in macrophages and Kupffer cells. The first one displayed the well-described alteration V162 Delta and the second a novel mutation (R178G) that was further detected in two relatives in a pedigree analysis. The proposed procedure would facilitate the wide-range molecular analysis of the SLC40A1 gene, contributing to better understanding the pathogenesis of the ferroportin disease.

  3. Impact of plasmids, including those encodingVirB4/D4 type IV secretion systems, on Salmonella enterica serovar Heidelberg virulence in macrophages and epithelial cells.

    Science.gov (United States)

    Gokulan, Kuppan; Khare, Sangeeta; Rooney, Anthony W; Han, Jing; Lynne, Aaron M; Foley, Steven L

    2013-01-01

    Salmonella enterica serovar Heidelberg (S. Heidelberg) can cause foodborne illness in humans following the consumption of contaminated meat and poultry products. Recent studies from our laboratory have demonstrated that certain S. Heidelberg isolated from food-animal sources harbor multiple transmissible plasmids with genes that encode antimicrobial resistance, virulence and a VirB4/D4 type-IV secretion system. This study examines the potential role of these transmissible plasmids in bacterial uptake and survival in intestinal epithelial cells and macrophages, and the molecular basis of host immune system modulation that may be associated with disease progression. A series of transconjugant and transformant strains were developed with different combinations of the plasmids to determine the roles of the individual and combinations of plasmids on virulence. Overall the Salmonella strains containing the VirB/D4 T4SS plasmids entered and survived in epithelial cells and macrophages to a greater degree than those without the plasmid, even though they carried other plasmid types. During entry in macrophages, the VirB/D4 T4SS encoding genes are up-regulated in a time-dependent fashion. When the potential mechanisms for increased virulence were examined using an antibacterial Response PCR Array, the strain containing the T4SS down regulated several host innate immune response genes which likely contributed to the increased uptake and survival within macrophages and epithelial cells.

  4. Brucella abortus virB12 is expressed during infection but is not an essential component of the type IV secretion system.

    Science.gov (United States)

    Sun, Yao-Hui; Rolán, Hortensia G; den Hartigh, Andreas B; Sondervan, David; Tsolis, Renée M

    2005-09-01

    The Brucella abortus virB operon, consisting of 11 genes, virB1 to virB11, and two putative genes, orf12 (virB12) and orf13, encodes a type IV secretion system (T4SS) that is required for intracellular replication and persistent infection in the mouse model. This study was undertaken to determine whether orf12 (virB12) encodes an essential part of the T4SS apparatus. The virB12 gene was found to encode a 17-kDa protein, which was detected in vitro in B. abortus grown to stationary phase. Mice infected with B. abortus 2308 produced an antibody response to the protein encoded by virB12, showing that this gene is expressed during infection. Expression of virB12 was not required for survival in J774 macrophages. VirB12 was also dispensable for the persistence of B. abortus, B. melitensis, and B. suis in mice up to 4 weeks after infection, since deletion mutants lacking virB12 were recovered from splenic tissue at wild-type levels. These results show that VirB12 is not essential for the persistence of the human-pathogenic Brucella spp. in the mouse and macrophage models of infection.

  5. Investigation of biofilm formation ability, antimicrobial resistance and the staphylococcal cassette chromosome mec patterns of methicillin resistant Staphylococcus epidermidis with different sequence types isolated from children.

    Science.gov (United States)

    Soroush, Setareh; Jabalameli, Fereshteh; Taherikalani, Morovat; Amirmozafari, Nour; Fooladi, Abbas Ali Imani; Asadollahi, Khairollah; Beigverdi, Reza; Emaneini, Mohammad

    2016-04-01

    This study investigated the molecular characterizations of 80 methicillin resistant Staphylococcus epidermidis (MRSE) collected during 2012-2013 in Tehran Children's Medical Center, Iran. About 90% of MRSE isolates were multi-drug resistant (MDR) and the highest resistance was observed to cotrimoxazole and they were quite sensitive to quinupristin-dalfopristin and linezolid. Though vanA gene was not detected, the majority of isolates showed intermediate resistance to vancomycin (MIC90 16 μg/ml). Resistance to mupirocin was observed in 18 isolates. Staphylococcal cassette chromosome mec (SCCmec) types V, III, IV and II were detected in 23.75%, 7.5%, 6.25% and 5% of isolates respectively, in some of which the additional parts of mec or ccr complexes were observed. In 57.5% MRSE isolates SCCmec types were not classified. 41.2% of MRSE isolates were carrying intercellular adhesion (ica) operon and 40% had strong or intermediate biofilm. The types of arginine catabolic mobile element (ACME) were limited to type I and II. Nine sequence types (STs) were seen in mupirocin resistant MRSE isolates. The common STs were ST2, ST5 and ST22 with 27.7% (5/18), 22.2% (4/18) and 16.6% (3/18) frequencies, respectively. ST23, ST54 and ST179 plus three novels STs 580, 581,588 were also observed. The majority of STs, 83.3% (15/18) belonged to clonal complex 2 (CC2). The spread of antibiotic resistance and virulence factors among MRSE species is an alarming sign in Children's Hospitals. The combination of these two issues leads to increase the chance of successfully establishing of common STs in hospital environments, and promotes the device-related infections and bacteremia. Copyright © 2016 Elsevier Ltd. All rights reserved.

  6. Transient Silencing of a Type IV P-Type ATPase, Atp10c, Results in Decreased Glucose Uptake in C2C12 Myotubes.

    Science.gov (United States)

    Hurst, S E; Minkin, S C; Biggerstaff, J; Dhar, M S

    2012-01-01

    Atp10c is a strong candidate gene for diet-induced obesity and type 2 diabetes. To identify molecular and cellular targets of ATP10C, Atp10c expression was altered in vitro in C2C12 skeletal muscle myotubes by transient transfection with an Atp10c-specific siRNA. Glucose uptake assays revealed that insulin stimulation caused a significant 2.54-fold decrease in 2-deoxyglucose uptake in transfected cells coupled with a significant upregulation of native mitogen-activated protein kinases (MAPKs), p38, and p44/42. Additionally, glucose transporter-1 (GLUT1) was significantly upregulated; no changes in glucose transporter-4 (GLUT4) expression were observed. The involvement of MAPKs was confirmed using the specific inhibitor SB203580, which downregulated the expression of native and phosphorylated MAPK proteins in transfected cells without any changes in insulin-stimulated glucose uptake. Results indicate that Atp10c regulates glucose metabolism, at least in part via the MAPK pathway, and, thus, plays a significant role in the development of insulin resistance and type 2 diabetes.

  7. Transient Silencing of a Type IV P-Type ATPase, Atp10c, Results in Decreased Glucose Uptake in C2C12 Myotubes

    Directory of Open Access Journals (Sweden)

    S. E. Hurst

    2012-01-01

    Full Text Available Atp10c is a strong candidate gene for diet-induced obesity and type 2 diabetes. To identify molecular and cellular targets of ATP10C, Atp10c expression was altered in vitro in C2C12 skeletal muscle myotubes by transient transfection with an Atp10c-specific siRNA. Glucose uptake assays revealed that insulin stimulation caused a significant 2.54-fold decrease in 2-deoxyglucose uptake in transfected cells coupled with a significant upregulation of native mitogen-activated protein kinases (MAPKs, p38, and p44/42. Additionally, glucose transporter-1 (GLUT1 was significantly upregulated; no changes in glucose transporter-4 (GLUT4 expression were observed. The involvement of MAPKs was confirmed using the specific inhibitor SB203580, which downregulated the expression of native and phosphorylated MAPK proteins in transfected cells without any changes in insulin-stimulated glucose uptake. Results indicate that Atp10c regulates glucose metabolism, at least in part via the MAPK pathway, and, thus, plays a significant role in the development of insulin resistance and type 2 diabetes.

  8. Ehlers-Danlos Syndrome type IV: a multi-exon deletion in one of the two COL3A1 alleles affecting structure, stability, and processing of type III procollagen

    Energy Technology Data Exchange (ETDEWEB)

    Superti-Furga, A.; Gugler, E.; Gitzelmann, R.; Steinmann, B.

    1988-05-05

    The authors have studied a patient with severe, dominantly inherited Ehlers-Danlos syndrome type IV. The results indicate that this patient carries a deletion of 3.3 kilobase pairs in the triple helical coding domain of one of the two alleles for the pro-..cap alpha..-chains of type III collagen (COL3A1). His cultured skin fibroblasts contain equal amounts of normal length mRNA and of mRNA shortened by approximately 600 bases, and synthesize both normal and shortened pro-..cap alpha..1(III)-chains. In procollagen molecules containing one or more shortened chains, a triple helix is formed with a length of only about 780 amino acids. The mutant procollagen molecules have decreased thermal stability, are less efficiently secreted, and are not processed as their normal counterpart. The deletion in this family is the first mutation to be described in COL3A1.

  9. Glycogen Storage Disease Type IV

    DEFF Research Database (Denmark)

    Bendroth-Asmussen, Lisa; Aksglaede, Lise; Gernow, Anne B

    2016-01-01

    A 30-yr-old woman presented with 2 consecutive miscarriages within 7 mo. Histopathologic examination of the placental tissue showed intracytoplasmic inclusion vacuoles with a strong reaction in Periodic acid-Schiff staining and a slightly pallor reaction in alcian blue staining. Additional...

  10. Membrane and core periplasmic Agrobacterium tumefaciens virulence Type IV secretion system components localize to multiple sites around the bacterial perimeter during lateral attachment to plant cells.

    Science.gov (United States)

    Aguilar, Julieta; Cameron, Todd A; Zupan, John; Zambryski, Patricia

    2011-01-01

    Type IV secretion systems (T4SS) transfer DNA and/or proteins into recipient cells. Here we performed immunofluorescence deconvolution microscopy to localize the assembled T4SS by detection of its native components VirB1, VirB2, VirB4, VirB5, VirB7, VirB8, VirB9, VirB10, and VirB11 in the C58 nopaline strain of Agrobacterium tumefaciens, following induction of virulence (vir) gene expression. These different proteins represent T4SS components spanning the inner membrane, periplasm, or outer membrane. Native VirB2, VirB5, VirB7, and VirB8 were also localized in the A. tumefaciens octopine strain A348. Quantitative analyses of the localization of all the above Vir proteins in nopaline and octopine strains revealed multiple foci in single optical sections in over 80% and 70% of the bacterial cells, respectively. Green fluorescent protein (GFP)-VirB8 expression following vir induction was used to monitor bacterial binding to live host plant cells; bacteria bind predominantly along their lengths, with few bacteria binding via their poles or subpoles. vir-induced attachment-defective bacteria or bacteria without the Ti plasmid do not bind to plant cells. These data support a model where multiple vir-T4SS around the perimeter of the bacterium maximize effective contact with the host to facilitate efficient transfer of DNA and protein substrates. Transfer of DNA and/or proteins to host cells through multiprotein type IV secretion system (T4SS) complexes that span the bacterial cell envelope is critical to bacterial pathogenesis. Early reports suggested that T4SS components localized at the cell poles. Now, higher-resolution deconvolution fluorescence microscopy reveals that all structural components of the Agrobacterium tumefaciens vir-T4SS, as well as its transported protein substrates, localize to multiple foci around the cell perimeter. These results lead to a new model of A. tumefaciens attachment to a plant cell, where A. tumefaciens takes advantage of the multiple

  11. The S(IV)-type Asteroids as Ordinary Chondrite Parent Body Candidates: Implications for the Completeness of the Meteorite Sample of Asteroids

    Science.gov (United States)

    Gaffey, M. J.

    1995-09-01

    The discrepancy between the abundance of ordinary chondrites (OCs) among the meteorites and the rarity of unambiguously similar assemblages in the asteroid belt has been a major point of discussion within and between the asteroid and meteorite communities. Various resolutions to this apparent paradox have been proposed [e.g., 1-5], including: 1) interpretations of S-type asteroid spectra are incorrect due to space weathering effects; 2) ordinary chondrites derive from a few rare but favorably situated parent bodies; 3) OCs come from a residual population of small unheated mainbelt asteroids; 4) shock effects darken OC parent body surfaces disguising them as C-type asteroids, and 5) OCs come from inner solar system planetesimals ejected to the Oort cloud which have been recently perturbed into Earth-crossing orbits. Although none of these possibilities has yet been rigorously excluded, recent investigations suggest that the resolution of the apparent paradox lies in some combination of the first three options. For option 3, the discovery of a small mainbelt asteroid with an OC-like spectrum indicates OC-assemblages among the smaller mainbelt asteroids [6], although their abundance is still low in the current sample [7]. For option 2, the mineralogical survey indicated that while most S-asteroids could be rigorously excluded on mineralogical criteria, the S(IV) subtype of this class has silicate compositions within the OC range [8]. The S(IV)-objects are concentrated near the 3:1 secular resonance at 2.5 AU providing an efficient escape into Earth-crossing orbits. Unfortunately for a simple resolution of the OC parent body question, S(IV) spectra still exhibit weaker silicate features and redder spectral slopes than OC assemblages. Although significant uncertainties remain, optical alteration of asteroid surfaces interpreted from the Galileo images of Ida and Gaspra may reconcile the mismatch between OC and S(IV) spectra [option 1]. Although only a subset of the S(IV

  12. Asteroids IV

    Science.gov (United States)

    Michel, Patrick; DeMeo, Francesca E.; Bottke, William F.

    . Asteroids, like planets, are driven by a great variety of both dynamical and physical mechanisms. In fact, images sent back by space missions show a collection of small worlds whose characteristics seem designed to overthrow our preconceived notions. Given their wide range of sizes and surface compositions, it is clear that many formed in very different places and at different times within the solar nebula. These characteristics make them an exciting challenge for researchers who crave complex problems. The return of samples from these bodies may ultimately be needed to provide us with solutions. In the book Asteroids IV, the editors and authors have taken major strides in the long journey toward a much deeper understanding of our fascinating planetary ancestors. This book reviews major advances in 43 chapters that have been written and reviewed by a team of more than 200 international authorities in asteroids. It is aimed to be as comprehensive as possible while also remaining accessible to students and researchers who are interested in learning about these small but nonetheless important worlds. We hope this volume will serve as a leading reference on the topic of asteroids for the decade to come. We are deeply indebted to the many authors and referees for their tremendous efforts in helping us create Asteroids IV. We also thank the members of the Asteroids IV scientific organizing committee for helping us shape the structure and content of the book. The conference associated with the book, "Asteroids Comets Meteors 2014" held June 30-July 4, 2014, in Helsinki, Finland, did an outstanding job of demonstrating how much progress we have made in the field over the last decade. We are extremely grateful to our host Karri Muinonnen and his team. The editors are also grateful to the Asteroids IV production staff, namely Renée Dotson and her colleagues at the Lunar and Planetary Institute, for their efforts, their invaluable assistance, and their enthusiasm; they made life as

  13. A type IV P-type ATPase affects insulin-mediated glucose uptake in adipose tissue and skeletal muscle in mice.

    Science.gov (United States)

    Dhar, Madhu S; Yuan, Joshua S; Elliott, Sarah B; Sommardahl, Carla

    2006-12-01

    Mice carrying two pink-eyed dilution (p) locus heterozygous deletions represent a novel polygenic mouse model of type 2 diabetes associated with obesity. Atp10c, a putative aminophospholipid transporter on mouse chromosome 7, is a candidate for the phenotype. The phenotype is diet-induced. As a next logical step in the validation and characterization of the model, experiments to analyze metabolic abnormalities associated with these mice were carried out. Results demonstrate that mutants (inheriting the p deletion maternally) heterozygous for Atp10c are hyperinsulinemic, insulin-resistant and have an altered insulin-stimulated response in peripheral tissues. Adipose tissue and the skeletal muscle are the targets, and GLUT4-mediated glucose uptake is the specific metabolic pathway associated with Atp10c deletion. Insulin resistance primarily affects the adipose tissue and the skeletal muscle, and the effect in the liver is secondary. Gene expression profiling using microarray and real-time PCR show significant changes in the expression of four genes--Vamp2, Dok1, Glut4 and Mapk14--involved in insulin signaling. The expression of Atp10c is also significantly altered in the adipose tissue and the soleus muscle. The most striking observation is the loss of Atp10c expression in the mutants, specifically in the soleus muscle, after eating the high-fat diet for 12 weeks. In conclusion, experiments suggest that the target genes and/or their cognate factors in conjunction with Atp10c presumably affect the normal translocation and sequestration of GLUT4 in both the target tissues.

  14. [Expression of contractile proteins alpha-actin and myosin of smooth muscle cells and collagen of IV type in human placenta at placental insufficiency in III trimester of pregnancy].

    Science.gov (United States)

    Khozhaĭ, L I; Otellin, V A; Pozharisskiĭ, K M; Pavlova, N G

    2010-01-01

    Changes of expression of contractile proteins (alpha-actin and myosin of smooth muscle cell) and of collagen of IV type in stroma of human placental villi were studied at the diagnosed placental insufficiency (PI) in III trimester of pregnancy. The study revealed pronounced disturbances of expression of contractile proteins and collagen of IV type at PI. It is shown that in perivascular envelopes of vessels of stem and intermediate villi there is present a much greater amount of cells expressing smooth muscle actin and myosin. These cells are arranged by the denser concentric layers and more compactly than in norm and fill the intervascular space inside the villi. The width of perivascular envelopes of vessels is higher, while vascular lumens are lower than in norm. In terminal villi the capillary walls are thickened and the number of pericytes immunopositive against the smooth muscle cell alpha-actin and myosin as well as collagen of IV type is increased. The change of synthesis of the cytoskeletal contractile proteins and collagen of IV type is shown to lead to structural disturbances of villi of different types and of perivascular areas and vessels, which doubtlessly indicates their participation in pathogenesis of placental dysfunction and of disturbance of placental hemodynamics.

  15. Study on observation condition and on simple evaluation method of TypeIV creep damage of Mod.9Cr-1Mo steel

    Science.gov (United States)

    Kawashima, F.; Masumitsu, T.; Negi, K.; Watanabe, K.; Fujiwara, K.

    2017-05-01

    Type IV creep damage is the phenomena that a large number of voids in micron size initiate, grow, coalesce each other and become large cracks. They initiate in welded joints of high chromium steel in power plant. The density of voids, the number of voids per a unit area, is used to evaluate the residual life. The observed density of voids depends on the observation conditions; the observation area and the magnification of observed photograph of metallographic structure, because voids do not distribute uniformly and the small magnification misses the small voids. In previous study, we studied the influence of them with simulated fine-grain HAZ of Mod.9Cr-1Mo steel, and proposed how to determine the appropriate observation area for the temporary allowable error. We also proposed the method to evaluate the start time of initiation of voids, the initiation rate of voids and the growth rate of voids based on the relation between the observed density of voids and the magnification. But the experimental data was short. In this paper we showed new data, but they were yet not sufficient. We used FEM analysis and considered why enough data had been not taken.

  16. Treatment of a direct carotid-cavernous fistula in a patient with type IV Ehlers-Danlos syndrome: a novel approach

    Energy Technology Data Exchange (ETDEWEB)

    Hollands, J.K.; Santarius, T.; Kirkpatrick, P.J. [Addenbrooke' s Hospital, Department of Neurosurgery, Cambridge (United Kingdom); Higgins, J.N. [Addenbrooke' s Hospital, Department of Neuroradiology, Cambridge (United Kingdom)

    2006-07-15

    We report a case of a 34-year-old female with type IV Ehlers-Danlos syndrome diagnosed with a carotid cavernous fistula presenting with progressive proptosis. Endovascular embolization using balloons or coils carries a high risk of complications in this group of patients, owing to the extreme fragility of the blood vessels. Initial treatment was conservative until an intracerebral haemorrhage occurred. To avoid transfemoral angiography, the ipsilateral carotid arteries and the internal jugular vein were surgically exposed for insertion of two endovascular sheaths. The patient was transferred from theatre to the angiography suite and the sheaths were used for embolization access. The fistula was closed, with preservation of the carotid artery, using Guglielmi detachable coils deployed in the cavernous sinus from the arterial and venous sides. Rapid resolution of symptoms and signs followed, which was sustained at 6-month follow-up. This technique offers alternative access for endovascular treatment, which may reduce the high incidence of mortality associated with catheter angiography in this condition. (orig.)

  17. Delineation of polar localization domains of Agrobacterium tumefaciens type IV secretion apparatus proteins VirB4 and VirB11.

    Science.gov (United States)

    Das, Aditi; Das, Anath

    2014-10-01

    Agrobacterium tumefaciens transfers DNA and proteins to a plant cell through a type IV secretion apparatus assembled by the VirB proteins. All VirB proteins localized to a cell pole, although these conclusions are in dispute. To study subcellular location of the VirB proteins and to identify determinants of their subcellular location, we tagged two proteins, VirB4 and VirB11, with the visual marker green fluorescent protein (GFP) and studied localization of the fusion proteins by epifluorescence microscopy. Both GFP-VirB4 and GFP-VirB11 fusions localized to a single cell pole. GFP-VirB11 was also functional in DNA transfer. To identify the polar localization domains (PLDs) of VirB4 and VirB11, we analyzed fusions of GFP with smaller segments of the two proteins. Two noncontiguous regions in VirB4, residues 236-470 and 592-789, contain PLDs. The VirB11 PLD mapped to a 69 amino acid segment, residues 149-217, in the central region of the protein. These domains are probably involved in interactions that target the two proteins to a cell pole. © 2014 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

  18. Enzyme-linked immunosorbent serum assay specific for the 7S domain of Collagen Type IV (P4NP 7S)

    DEFF Research Database (Denmark)

    Leeming, Diana J; Nielsen, Mette J; Dai, Yueqin

    2012-01-01

    Aim:  The present study describes the ability of a newly developed N-terminal pro-peptides of type IV collagen 7S domain (P4NP 7S) competitive enzyme-linked immunosorbent assay (ELISA) for describing liver fibrosis. The assay applies a monoclonal antibody specific for a PIVNP 7S epitope 100......% homologous in the human, rat, and mouse species. Methods:  Monoclonal antibodies were raised against selected P4NP 7S specific sequences. Antibodies were screened and a competitive ELISA assay was developed using a selected antibody. The assay was evaluated in relation to technical performance, and in two...... preclinical liver fibrosis models; the bile duct ligation model (BDL) and the carbon tetrachloride model (CCL4) both performed in rats. Results:  A technically robust P4NP 7S ELISA assay using a monoclonal antibody was produced. In the BDL and CCL4 liver fibrosis models it was observed that the P4NP 7S levels...

  19. Prevalence and molecular typing of the antiseptic resistance genes ...

    African Journals Online (AJOL)

    Additionally, two predominant PFGE (B and A) and spa types (t037 and t042) were identified along with two major antimicrobial resistance profiles. All of these qacA/B-positive strains strains were pvl-negative by PCR. The qacA/B-positive MRSA strains all contained the group III SCCmec element. These strains were ...

  20. Factors associated with the support of pricking (female genital cutting type IV) among Somali immigrants - a cross-sectional study in Sweden.

    Science.gov (United States)

    Wahlberg, Anna; Johnsdotter, Sara; Ekholm Selling, Katarina; Källestål, Carina; Essén, Birgitta

    2017-08-08

    Pricking, classified as female genital cutting (FGC) type IV by the World Health Organization, is an under-researched area gaining momentum among diaspora communities. Our aim was to explore factors associated with being supportive of pricking among Somalis in Sweden. In a cross-sectional design, attitudes and knowledge regarding FGC, and measures of socioeconomic status, acculturation, and social capital, were assessed by a 49-item questionnaire in four municipalities in Sweden. Data were collected in 2015 from 648 Somali men and women, ≥ 18 years old, of which 113 supported the continuation of pricking. Logistic regression was used for the analysis. Those more likely to support the continuation of pricking were older, originally from rural areas, and newly arrived in Sweden. Further, those who reported that they thought pricking was: acceptable, according to their religion (aOR: 10.59, 95% CI: 5.44-20.62); not a violation of children's rights (aOR: 2.86, 95% CI: 1.46-5.61); and did not cause long-term health complications (aOR: 5.52, 95% CI: 2.25-13.52) had higher odds of supporting pricking. Religion was strongly associated with the support of pricking among both genders. However, for men, children's rights and the definition of pricking as FGC or not were important aspects in how they viewed pricking, while, for women, health complications and respectability were important. Values known to be associated with FGC in general are also related to pricking. Hence, there seems to be a change in what types of FGC are supported rather than in their perceived values.

  1. Community-acquired necrotizing pneumonia caused by methicillin-resistant Staphylococcus aureus ST30-SCCmecIVc-spat019-PVL positive in San Antonio de Areco, Argentina

    Directory of Open Access Journals (Sweden)

    Silvina Fernández

    2015-03-01

    Full Text Available Community-acquired methicillin-resistant Staphylococcus aureus is the first cause of skin and soft tissue infections, but can also produce severe diseases such as bacteremia, osteomyelitis and necrotizing pneumonia. Some S. aureus lineages have been described in cases of necrotizing pneumonia worldwide, usually in young, previously healthy patients. In this work, we describe a fatal case of necrotizing pneumonia due to community-acquired methicillin-resistant S. aureus clone ST30-SCCmecIVc-spat019-PVL positive in an immunocompetent adult patient.

  2. Single-catheter approach for ablation of the slow pathway in a patient with type IV Ehlers-Danlos syndrome and AV nodal reentrant tachycardia using a magnetic navigation system

    NARCIS (Netherlands)

    T. Szili-Torok (Tamas); E. Jessurun; L.J.L.M. Jordaens (Luc)

    2008-01-01

    textabstractPatients with Ehlers-Danlos syndrome type IV have thin-walled, friable arteries and veins. Invasive procedures carry a significantly increased risk for perforation of blood vessels. The aim of this case report is to demonstrate the feasibility and potential benefits of using a

  3. Identification of VceA and VceC, two members of the VjbR regulon that are translocated into macrophages by the Brucella type IV secretion system

    NARCIS (Netherlands)

    de Jong, Maarten F.; Sun, Yao-Hui; den Hartigh, Andreas B.; van Dijl, Jan Maarten; Tsolis, Renee M.

    2008-01-01

    Survival and replication inside host cells by Brucella spp. requires a type IV secretion system (T4SS), encoded by the virB locus. However, the identity of the molecules secreted by the T4SS has remained elusive. We hypothesized that proteins translocated by the T4SS would be co-regulated with the

  4. Dissemination of multiple MRSA clones among community-associated methicillin-resistant Staphylococcus aureus infections from Japanese children with impetigo.

    Science.gov (United States)

    Hisata, Ken; Ito, Teruyo; Matsunaga, Nobuaki; Komatsu, Mitsutaka; Jin, Jingxun; Li, Shanshuang; Watanabe, Shinya; Shimizu, Toshiaki; Hiramatsu, Keiichi

    2011-10-01

    The proportion of MRSA strains that cause skin and soft infections has recently increased. In 3 months we have characterized 17 MRSA strains isolated from children with impetigo at a Japanese hospital. Seventeen MRSA strains belonged to 7 clones defined by clonal complex (CC) in MLST genotype and type of SCCmec, which were rarely identified among healthcare-associated MRSA: CC 91-SCCmecIIb (4 strains); CC91-SCCmecIIn (2 strains); CC91-SCCmecIVa (2 strains); CC91-SCCmecV (4 strains); CC88-SCCmecIVg (3 strains); CC1-SCCmecIVc (1 strain); and CC5-SCCmecIVn (1 strain). Although one strain belonged to CC5, which has been commonly identified in healthcare-associated MRSA, it did not carry type II SCCmec, but carried type IV SCCmec. Fourteen of the 17 strains carried exfoliative toxin a or b gene, and none carried Panton-Valentine leukocidine gene. Furthermore, we determined the entire nucleotide sequences of two type V SCCmec elements carried by strains JCSC5952, a CC91 strain, and TSGH17, a Taiwanese CC59 strain. The structure of SCCmecJCSC5952 was more than 99% homologous in nucleotide identity with those of Taiwanese PVL-positive ST59 MRSA strains TSGH17 and PM1, which were designated as type V (5C2&5). Identification of multiple MRSA clones distinct from those disseminating at the hospital suggests that MRSA strains might be emerging in the community from MSSA strains by acquiring SCCmec elements on various occasions. Carriage of the similar type V(5C2&5) SCCmec element by strains of distinct genetic backgrounds, CC91 and CC59, suggested horizontal transfer of the SCCmec element.

  5. Association of DSM-IV Posttraumatic Stress Disorder With Traumatic Experience Type and History in the World Health Organization World Mental Health Surveys.

    Science.gov (United States)

    Liu, Howard; Petukhova, Maria V; Sampson, Nancy A; Aguilar-Gaxiola, Sergio; Alonso, Jordi; Andrade, Laura Helena; Bromet, Evelyn J; de Girolamo, Giovanni; Haro, Josep Maria; Hinkov, Hristo; Kawakami, Norito; Koenen, Karestan C; Kovess-Masfety, Viviane; Lee, Sing; Medina-Mora, Maria Elena; Navarro-Mateu, Fernando; O'Neill, Siobhan; Piazza, Marina; Posada-Villa, José; Scott, Kate M; Shahly, Victoria; Stein, Dan J; Ten Have, Margreet; Torres, Yolanda; Gureje, Oye; Zaslavsky, Alan M; Kessler, Ronald C

    2017-03-01

    Previous research has documented significant variation in the prevalence of posttraumatic stress disorder (PTSD) depending on the type of traumatic experience (TE) and history of TE exposure, but the relatively small sample sizes in these studies resulted in a number of unresolved basic questions. To examine disaggregated associations of type of TE history with PTSD in a large cross-national community epidemiologic data set. The World Health Organization World Mental Health surveys assessed 29 TE types (lifetime exposure, age at first exposure) with DSM-IV PTSD that was associated with 1 randomly selected TE exposure (the random TE) for each respondent. Surveys were administered in 20 countries (n = 34 676 respondents) from 2001 to 2012. Data were analyzed from October 1, 2015, to September 1, 2016. Prevalence of PTSD assessed with the Composite International Diagnostic Interview. Among the 34 676 respondents (55.4% [SE, 0.6%] men and 44.6% [SE, 0.6%] women; mean [SE] age, 43.7 [0.2] years), lifetime TE exposure was reported by a weighted 70.3% of respondents (mean [SE] number of exposures, 4.5 [0.04] among respondents with any TE). Weighted (by TE frequency) prevalence of PTSD associated with random TEs was 4.0%. Odds ratios (ORs) of PTSD were elevated for TEs involving sexual violence (2.7; 95% CI, 2.0-3.8) and witnessing atrocities (4.2; 95% CI, 1.0-17.8). Prior exposure to some, but not all, same-type TEs was associated with increased vulnerability (eg, physical assault; OR, 3.2; 95% CI, 1.3-7.9) or resilience (eg, participation in sectarian violence; OR, 0.3; 95% CI, 0.1-0.9) to PTSD after the random TE. The finding of earlier studies that more general history of TE exposure was associated with increased vulnerability to PTSD across the full range of random TE types was replicated, but this generalized vulnerability was limited to prior TEs involving violence, including participation in organized violence (OR, 1.3; 95% CI, 1.0-1.6), experience of

  6. Cyclic Di-GMP Binding by an Assembly ATPase (PilB2) and Control of Type IV Pilin Polymerization in the Gram-Positive Pathogen Clostridium perfringens.

    Science.gov (United States)

    Hendrick, William A; Orr, Mona W; Murray, Samantha R; Lee, Vincent T; Melville, Stephen B

    2017-05-15

    The Gram-positive pathogen Clostridium perfringens possesses type IV pili (TFP), which are extracellular fibers that are polymerized from a pool of pilin monomers in the cytoplasmic membrane. Two proteins that are essential for pilus functions are an assembly ATPase (PilB) and an inner membrane core protein (PilC). Two homologues each of PilB and PilC are present in C. perfringens, called PilB1/PilB2 and PilC1/PilC2, respectively, along with four pilin proteins, PilA1 to PilA4. The gene encoding PilA2, which is considered the major pilin based on previous studies, is immediately downstream of the pilB2 and pilC2 genes. Purified PilB2 had ATPase activity, bound zinc, formed hexamers even in the absence of ATP, and bound the second messenger molecule cyclic di-GMP (c-di-GMP). Circular dichroism spectroscopy of purified PilC2 indicated that it retained its predicted degree of alpha-helical secondary structure. Even though no direct interactions between PilB2 and PilC2 could be detected in vivo or in vitro even in the presence of c-di-GMP, high levels of expression of a diguanylate cyclase from C. perfringens (CPE1788) stimulated polymerization of PilA2 in a PilB2- and PilC2-dependent manner. These results suggest that PilB2 activity is controlled by c-di-GMP levels in vivo but that PilB2-PilC2 interactions are either transitory or of low affinity, in contrast to results reported previously from in vivo studies of the PilB1/PilC1 pair in which PilC1 was needed for polar localization of PilB1. This is the first biochemical characterization of a c-di-GMP-dependent assembly ATPase from a Gram-positive bacterium.IMPORTANCE Type IV pili (TFP) are protein fibers involved in important bacterial functions, including motility, adherence to surfaces and host cells, and natural transformation. All clostridia whose genomes have been sequenced show evidence of the presence of TFP. The genetically tractable species Clostridium perfringens was used to study proteins involved in

  7. SDSS IV MaNGA: Deep observations of extra-planar, diffuse ionized gas around late-type galaxies from stacked IFU spectra

    Science.gov (United States)

    Jones, A.; Kauffmann, G.; D'Souza, R.; Bizyaev, D.; Law, D.; Haffner, L.; Bahé, Y.; Andrews, B.; Bershady, M.; Brownstein, J.; Bundy, K.; Cherinka, B.; Diamond-Stanic, A.; Drory, N.; Riffel, R. A.; Sánchez, S. F.; Thomas, D.; Wake, D.; Yan, R.; Zhang, K.

    2017-03-01

    We have conducted a study of extra-planar diffuse ionized gas using the first year data from the MaNGA IFU survey. We have stacked spectra from 49 edge-on, late-type galaxies as a function of distance from the midplane of the galaxy. With this technique we can detect the bright emission lines Hα, Hβ, [O II]λλ3726, 3729, [O III]λ5007, [N II]λλ6549, 6584, and [S II]λλ6717, 6731 out to about 4 kpc above the midplane. With 16 galaxies we can extend this analysis out to about 9 kpc, I.e. a distance of 2Re, vertically from the midplane. In the halo, the surface brightnesses of the [O II] and Hα emission lines are comparable, unlike in the disk where Hα dominates. When we split the sample by specific star-formation rate, concentration index, and stellar mass, each subsample's emission line surface brightness profiles and ratios differ, indicating that extra-planar gas properties can vary. The emission line surface brightnesses of the gas around high specific star-formation rate galaxies are higher at all distances, and the line ratios are closer to ratios characteristic of H II regions compared with low specific star-formation rate galaxies. The less concentrated and lower stellar mass samples exhibit line ratios that are more like H II regions at larger distances than their more concentrated and higher stellar mass counterparts. The largest difference between different subsamples occurs when the galaxies are split by stellar mass. We additionally infer that gas far from the midplane in more massive galaxies has the highest temperatures and steepest radial temperature gradients based on their [N II]/Hα and [O II]/Hα ratios between the disk and the halo. SDSS IV.

  8. Evolutionary Dynamics of Pathoadaptation Revealed by Three Independent Acquisitions of the VirB/D4 Type IV Secretion System in Bartonella.

    Science.gov (United States)

    Harms, Alexander; Segers, Francisca H I D; Quebatte, Maxime; Mistl, Claudia; Manfredi, Pablo; Körner, Jonas; Chomel, Bruno B; Kosoy, Michael; Maruyama, Soichi; Engel, Philipp; Dehio, Christoph

    2017-03-01

    The α-proteobacterial genus Bartonella comprises a group of ubiquitous mammalian pathogens that are studied as a model for the evolution of bacterial pathogenesis. Vast abundance of two particular phylogenetic lineages of Bartonella had been linked to enhanced host adaptability enabled by lineage-specific acquisition of a VirB/D4 type IV secretion system (T4SS) and parallel evolution of complex effector repertoires. However, the limited availability of genome sequences from one of those lineages as well as other, remote branches of Bartonella has so far hampered comprehensive understanding of how the VirB/D4 T4SS and its effectors called Beps have shaped Bartonella evolution. Here, we report the discovery of a third repertoire of Beps associated with the VirB/D4 T4SS of B. ancashensis, a novel human pathogen that lacks any signs of host adaptability and is only distantly related to the two species-rich lineages encoding a VirB/D4 T4SS. Furthermore, sequencing of ten new Bartonella isolates from under-sampled lineages enabled combined in silico analyses and wet lab experiments that suggest several parallel layers of functional diversification during evolution of the three Bep repertoires from a single ancestral effector. Our analyses show that the Beps of B. ancashensis share many features with the two other repertoires, but may represent a more ancestral state that has not yet unleashed the adaptive potential of such an effector set. We anticipate that the effectors of B. ancashensis will enable future studies to dissect the evolutionary history of Bartonella effectors and help unraveling the evolutionary forces underlying bacterial host adaptation. © The Author(s) 2017. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

  9. The conjugation protein TcpC from Clostridium perfringens is structurally related to the type IV secretion system protein VirB8 from Gram-negative bacteria.

    Science.gov (United States)

    Porter, Corrine J; Bantwal, Radhika; Bannam, Trudi L; Rosado, Carlos J; Pearce, Mary C; Adams, Vicki; Lyras, Dena; Whisstock, James C; Rood, Julian I

    2012-01-01

    Bacterial conjugation is important for the acquisition of virulence and antibiotic resistance genes. We investigated the mechanism of conjugation in Gram-positive pathogens using a model plasmid pCW3 from Clostridium perfringens. pCW3 encodes tetracycline resistance and contains the tcp locus, which is essential for conjugation. We showed that the unique TcpC protein (359 amino acids, 41 kDa) was required for efficient conjugative transfer, localized to the cell membrane independently of other conjugation proteins, and that membrane localization was important for its function, oligomerization and interaction with the conjugation proteins TcpA, TcpH and TcpG. The crystal structure of the C-terminal component of TcpC (TcpC(99-359)) was determined to 1.8-Å resolution. TcpC(99-359) contained two NTF2-like domains separated by a short linker. Unexpectedly, comparative structural analysis showed that each of these domains was structurally homologous to the periplasmic region of VirB8, a component of the type IV secretion system from Agrobacterium tumefaciens. Bacterial two-hybrid studies revealed that the C-terminal domain was critical for interactions with other conjugation proteins. The N-terminal region of TcpC was required for efficient conjugation, oligomerization and protein-protein interactions. We conclude that by forming oligomeric complexes, TcpC contributes to the stability and integrity of the conjugation apparatus, facilitating efficient pCW3 transfer. © 2011 Blackwell Publishing Ltd.

  10. Development and application of pathovar-specific monoclonal antibodies that recognize the lipopolysaccharide O antigen and the type IV fimbriae of Xanthomonas hyacinthi

    Energy Technology Data Exchange (ETDEWEB)

    Doorn, J. van; Ojanen-Reuhs, T.; Hollinger, T.C.; Reuhs, B.L.; Schots, A.; Boonekamp, P.M.; Oudega, B.

    1999-09-01

    The objective of this study was to develop a specific immunological diagnostic assay for yellow disease in hyacinths, using monoclonal antibodies (MAbs). Mice were immunized with a crude cell wall preparation (shear fraction) from Xanthomonas hyacinthi and with purified type IV fimbriae. Hybridomas were screened for a positive reaction with X. hyacinthi cells or fimbriae and for a negative reaction with X. translucens pv. graminis or Erwinia carotovora subsp. carotovora. Nine MAbs recognized fimbrial epitopes, as shown by immunoblotting, immunofluorescence, enzyme-linked immunosorbent assay (ELISA), and immunoelectron microscopy; however, three of these MAbs had weak cross-reactions with two X. translucens pathovars in immunoblotting experiments. Seven MAbs reacted with lipopolysaccharides and yielded a low-mobility ladder pattern on immunoblots. Subsequent analysis of MAb 2E5 showed that it specifically recognized an epitope on the O antigen, which was found to consist of rhamnose and fucose in a 2:1 molar ratio. The cross-reaction of MAb 2E5 with all X. hyacinthi strains tested showed that this O antigen is highly conserved within this species. MAb 1B10 also reacted with lipopolysaccharides. MAbs 2E5 and 1B10 were further tested in ELISA and immunoblotting experiments with cells and extracts from other pathogens. No cross-reaction was found with 27 other Xanthomonas pathovars tested or with 14 other bacterial species from other genera, such as Erwinia and Pseudomonas, indicating the high specificity of these antibodies. MAbs 2E5 and 1B10 were shown to be useful in ELISA for the detection of X. hyacinthi in infected hyacinths.

  11. A chromosomally located traHIJKCLMN operon encoding a putative type IV secretion system is involved in the virulence of Yersinia ruckeri.

    Science.gov (United States)

    Méndez, J; Fernández, L; Menéndez, A; Reimundo, P; Pérez-Pascual, D; Navais, R; Guijarro, J A

    2009-02-01

    Nucleotide sequence analysis of the region surrounding the pIVET8 insertion site in Yersinia ruckeri 150RiviXII, previously selected by in vivo expression technology (IVET), revealed the presence of eight genes (traHIJKCLMN [hereafter referred to collectively as the tra operon or tra cluster]), which are similar both in sequence and organization to the tra operon cluster found in the virulence-related plasmid pADAP from Serratia entomophila. Interestingly, the tra cluster of Y. ruckeri is chromosomally encoded, and no similar tra cluster has been identified yet in the genomic analysis of human pathogenic yersiniae. A traI insertional mutant was obtained by homologous recombination. Coinfection experiments with the mutant and the parental strain, as well as 50% lethal dose determinations, indicate that this operon is involved in the virulence of this bacterium. All of these results suggest the implication of the tra cluster in a virulence-related type IV secretion/transfer system. Reverse transcriptase PCR studies showed that this cluster is transcribed as an operon from a putative promoter located upstream of traH and that the mutation of traI had a polar effect. A traI::lacZY transcriptional fusion displayed higher expression levels at 18 degrees C, the temperature of occurrence of the disease, and under nutrient-limiting conditions. PCR detection analysis indicated that the tra cluster is present in 15 Y. ruckeri strains from different origins and with different plasmid profiles. The results obtained in the present study support the conclusion, already suggested by different authors, that Y. ruckeri is a very homogeneous species that is quite different from the other members of the genus Yersinia.

  12. Effector protein translocation by the Coxiella burnetii Dot/Icm type IV secretion system requires endocytic maturation of the pathogen-occupied vacuole.

    Directory of Open Access Journals (Sweden)

    Hayley J Newton

    Full Text Available The human pathogen Coxiella burnetii encodes a type IV secretion system called Dot/Icm that is essential for intracellular replication. The Dot/Icm system delivers bacterial effector proteins into the host cytosol during infection. The effector proteins delivered by C. burnetii are predicted to have important functions during infection, but when these proteins are needed during infection has not been clearly defined. Here, we use a reporter system consisting of fusion proteins that have a β-lactamase enzyme (BlaM fused to C. burnetii effector proteins to study protein translocation by the Dot/Icm system. Translocation of BlaM fused to the effector proteins CBU0077, CBU1823 and CBU1524 was not detected until 8-hours after infection of HeLa cells, which are permissive for C. burnetii replication. Translocation of these effector fusion proteins by the Dot/Icm system required acidification of the Coxiella-containing vacuole. Silencing of the host genes encoding the membrane transport regulators Rab5 or Rab7 interfered with effector translocation, which indicates that effectors are not translocated until bacteria traffic to a late endocytic compartment in the host cell. Similar requirements for effector translocation were discerned in bone marrow macrophages derived from C57BL/6 mice, which are primary cells that restrict the intracellular replication of C. burnetii. In addition to requiring endocytic maturation of the vacuole for Dot/Icm-mediated translocation of effectors, bacterial transcription was required for this process. Thus, translocation of effector proteins by the C. burnetii Dot/Icm system occurs after acidification of the CCV and maturation of this specialized organelle to a late endocytic compartment. This indicates that creation of the specialized vacuole in which C. burnetii replicates represents a two-stage process mediated initially by host factors that regulate endocytic maturation and then by bacterial effectors delivered into

  13. The Coxiella burnetii Dot/Icm System Delivers a Unique Repertoire of Type IV Effectors into Host Cells and Is Required for Intracellular Replication

    Science.gov (United States)

    Lührmann, Anja; Roy, Craig R.

    2011-01-01

    Coxiella burnetii, the causative agent of human Q fever, is an intracellular pathogen that replicates in an acidified vacuole derived from the host lysosomal network. This pathogen encodes a Dot/Icm type IV secretion system that delivers bacterial proteins called effectors to the host cytosol. To identify new effector proteins, the functionally analogous Legionella pneumophila Dot/Icm system was used in a genetic screen to identify fragments of C. burnetii genomic DNA that when fused to an adenylate cyclase reporter were capable of directing Dot/Icm-dependent translocation of the fusion protein into mammalian host cells. This screen identified Dot/Icm effectors that were proteins unique to C. burnetii, having no overall sequence homology with L. pneumophila Dot/Icm effectors. A comparison of C. burnetii genome sequences from different isolates revealed diversity in the size and distribution of the genes encoding many of these effectors. Studies examining the localization and function of effectors in eukaryotic cells provided evidence that several of these proteins have an affinity for specific host organelles and can disrupt cellular functions. The identification of a transposon insertion mutation that disrupts the dot/icm locus was used to validate that this apparatus was essential for translocation of effectors. Importantly, this C. burnetii Dot/Icm-deficient mutant was found to be defective for intracellular replication. Thus, these data indicate that C. burnetii encodes a unique subset of bacterial effector proteins translocated into host cells by the Dot/Icm apparatus, and that the cumulative activities exerted by these effectors enables C. burnetii to successfully establish a niche inside mammalian cells that supports intracellular replication. PMID:21637816

  14. Iron Starvation Conditions Upregulate Ehrlichia ruminantium Type IV Secretion System, tr1 Transcription Factor and map1 Genes Family through the Master Regulatory Protein ErxR

    Directory of Open Access Journals (Sweden)

    Amal Moumène

    2018-01-01

    Full Text Available Ehrlichia ruminantium is an obligatory intracellular bacterium that causes heartwater, a fatal disease in ruminants. Due to its intracellular nature, E. ruminantium requires a set of specific virulence factors, such as the type IV secretion system (T4SS, and outer membrane proteins (Map proteins in order to avoid and subvert the host's immune response. Several studies have been conducted to understand the regulation of the T4SS or outer membrane proteins, in Ehrlichia, but no integrated approach has been used to understand the regulation of Ehrlichia pathogenicity determinants in response to environmental cues. Iron is known to be a key nutrient for bacterial growth both in the environment and within hosts. In this study, we experimentally demonstrated the regulation of virB, map1, and tr1 genes by the newly identified master regulator ErxR (for Ehrlichia ruminantium expression regulator. We also analyzed the effect of iron depletion on the expression of erxR gene, tr1 transcription factor, T4SS and map1 genes clusters in E. ruminantium. We show that exposure of E. ruminantium to iron starvation induces erxR and subsequently tr1, virB, and map1 genes. Our results reveal tight co-regulation of T4SS and map1 genes via the ErxR regulatory protein at the transcriptional level, and, for the first time link map genes to the virulence function sensu stricto, thereby advancing our understanding of Ehrlichia's infection process. These results suggest that Ehrlichia is able to sense changes in iron concentrations in the environment and to regulate the expression of virulence factors accordingly.

  15. Transcription and translation products of the cytolysin gene psm-mec on the mobile genetic element SCCmec regulate Staphylococcus aureus virulence.

    Directory of Open Access Journals (Sweden)

    Chikara Kaito

    Full Text Available The F region downstream of the mecI gene in the SCCmec element in hospital-associated methicillin-resistant Staphylococcus aureus (HA-MRSA contains two bidirectionally overlapping open reading frames (ORFs, the fudoh ORF and the psm-mec ORF. The psm-mec ORF encodes a cytolysin, phenol-soluble modulin (PSM-mec. Transformation of the F region into the Newman strain, which is a methicillin-sensitive S. aureus (MSSA strain, or into the MW2 (USA400 and FRP3757 (USA300 strains, which are community-acquired MRSA (CA-MRSA strains that lack the F region, attenuated their virulence in a mouse systemic infection model. Introducing the F region to these strains suppressed colony-spreading activity and PSMα production, and promoted biofilm formation. By producing mutations into the psm-mec ORF, we revealed that (i both the transcription and translation products of the psm-mec ORF suppressed colony-spreading activity and promoted biofilm formation; and (ii the transcription product of the psm-mec ORF, but not its translation product, decreased PSMα production. These findings suggest that both the psm-mec transcript, acting as a regulatory RNA, and the PSM-mec protein encoded by the gene on the mobile genetic element SCCmec regulate the virulence of Staphylococcus aureus.

  16. Sorption of Th (IV) to silica as a function of pH, humic/fulvic acid, ionic strength, electrolyte type

    Energy Technology Data Exchange (ETDEWEB)

    Chen Changlun [Institute of Plasma Physics, Chinese Academy of Sciences, P.O. Box 1126, Hefei, 230031, Anhui (China)]. E-mail: clchen@ipp.ac.cn; Wang Xiangke [Institute of Plasma Physics, Chinese Academy of Sciences, P.O. Box 1126, Hefei, 230031, Anhui (China)]. E-mail: xkwang@ipp.ac.cn

    2007-02-15

    The removal behavior of thorium (Th(IV)) has been investigated in multicomponent systems containing silica (SiO{sub 2}) as the model of inorganic particles because of its widespread presence in the earth's crust and soil humic acid (HA)/fulvic acid (FA) by batch experiments. The influence of pH from 2 to 12, ionic strength from 0.02 to 0.2 M KNO{sub 3}, soil HA/FA concentration from 8.3 to 22.5 mg/L, and foreign cations (Li{sup +}, Na{sup +}, K{sup +}) and anions (NO{sub 3} {sup -}, Cl{sup -}) on the sorption of Th(IV) onto SiO{sub 2} was also tested. The sorption isotherms of Th(IV) at approximately constant pH (3.50{+-}0.02) were determined and analyzed regressively with three kinds of sorption isotherm models, i.e., linear, Langmuir, and Freundlich models. The results demonstrated that the sorption of Th(IV) onto SiO{sub 2} increased steeply with increasing pH from 2 to 4. Generally, humic substances (HSs) were shown to enhance Th(IV) sorption at low pH, but to reduce Th(IV) sorption at intermediate and high pH. It was a hypothesis that the significantly positive influence of HA/FA at pH from 2 to 4 on the sorption of Th(IV) onto SiO{sub 2} was attributed to strong surface binding of HA/FA on SiO{sub 2} and subsequently the formation of ternary surface complexes such as {identical_to}MO-O-HA-Th or {identical_to}MO-O-FA-Th. The results also demonstrated that the sorption was strongly dependent on the concentration of HA/FA, and independent of ionic strength and foreign ions under our experimental conditions.

  17. Helicobacter pylori exploits a unique repertoire of type IV secretion system components for pilus assembly at the bacteria-host cell interface.

    Directory of Open Access Journals (Sweden)

    Carrie L Shaffer

    2011-09-01

    Full Text Available Colonization of the human stomach by Helicobacter pylori is an important risk factor for development of gastric cancer. The H. pylori cag pathogenicity island (cag PAI encodes components of a type IV secretion system (T4SS that translocates the bacterial oncoprotein CagA into gastric epithelial cells, and CagL is a specialized component of the cag T4SS that binds the host receptor α5β1 integrin. Here, we utilized a mass spectrometry-based approach to reveal co-purification of CagL, CagI (another integrin-binding protein, and CagH (a protein with weak sequence similarity to CagL. These three proteins are encoded by contiguous genes in the cag PAI, and are detectable on the bacterial surface. All three proteins are required for CagA translocation into host cells and H. pylori-induced IL-8 secretion by gastric epithelial cells; however, these proteins are not homologous to components of T4SSs in other bacterial species. Scanning electron microscopy analysis reveals that these proteins are involved in the formation of pili at the interface between H. pylori and gastric epithelial cells. ΔcagI and ΔcagL mutant strains fail to form pili, whereas a ΔcagH mutant strain exhibits a hyperpiliated phenotype and produces pili that are elongated and thickened compared to those of the wild-type strain. This suggests that pilus dimensions are regulated by CagH. A conserved C-terminal hexapeptide motif is present in CagH, CagI, and CagL. Deletion of these motifs results in abrogation of CagA translocation and IL-8 induction, and the C-terminal motifs of CagI and CagL are required for formation of pili. In summary, these results indicate that CagH, CagI, and CagL are components of a T4SS subassembly involved in pilus biogenesis, and highlight the important role played by unique constituents of the H. pylori cag T4SS.

  18. Continued expansion of USA300-like methicillin-resistant Staphylococcus aureus (MRSA) among hospitalized patients in the United States.

    Science.gov (United States)

    Tickler, Isabella A; Goering, Richard V; Mediavilla, Jose R; Kreiswirth, Barry N; Tenover, Fred C

    2017-08-01

    We characterized spa types, SCCmec types, and antimicrobial resistance patterns of 516 methicillin-resistant Staphylococcus aureus (MRSA) isolates, collected between 2011 and 2014 from nares and blood cultures of United States patients. Among nares isolates, 45 spa types were observed; 29.9% were t002/SCCmec II and 30.9% were t008/SCCmec IV. Among blood isolates, 40 spa types were identified; 24.4% were t002/SCCmec II and 39.9% were type t008/SCCmec IV. Compared to data from our 2009-2010 survey, the percentage of t008/SCCmec IV isolates from nares increased significantly (20.4%-30.9%; P=0.004) while the percentage from positive blood cultures remained similar (39.2% versus 39.9%; P=0.921). There were also significant changes in the overall antimicrobial resistance patterns observed, including the decrease of the clindamycin, erythromycin, levofloxacin and moxifloxacin multidrug resistance pattern, likely the result of t002/SCCmec II strains being displaced by t008/SCCmec IV strains. Rates of high-level mupirocin resistance did not change significantly from our past study (4.1% compared to 4.7%; P=0.758) but an increase in low-level resistance, particularly among t002/SCCmec II isolates, was observed. Copyright © 2017 Elsevier Inc. All rights reserved.

  19. Substitution of Aspartate for glycine 1018 in the Type III procollagen (COL3AI) gene causes type IV Ehlers-Danlos Syndrome: The mutated allele is present in most blood leukocytes of the asymptomatic and mosaic mother

    Energy Technology Data Exchange (ETDEWEB)

    Kontusaari, S.; Tromp, G.; Kuivaniemi, H.; Prockop, D.J. (Thomas Jefferson Univ., Philadelphia, PA (United States)); Stolle, C. (Robert Wood Johnson Medical School, Piscataway, NJ (United States)); Pope, F.M.

    1992-09-01

    A proband with arterial ruptures and skin changes characteristic of the type IV variant of Ehlers-Danlos syndrome was found to have a single-base mutation in the type III procollagen gene, which converted the codon for glycine at amino position 1018 to a codon for aspartate. (Amino acid positions are numbered by the standard convention in which the first glycine of the triple-helical domain of an [alpha] chain is number 1. The numbers of positions in the [alpha]1(III) chains can be converted to positions in the human pro[alpha](III) chain by adding 167.). Nucleotide sequencing of overlapping PCR products in which the two alleles were distinguished demonstrated that the mutation of glycine 1018 was the only mutation that changed the primary structure of type III procollagen. The glycine substitution markedly decreased the amount of type III procollagen secreted into the medium by cultured skin fibroblasts from the proband. It is surprising that the same mutation was found in about 94% of the peripheral blood leukocytes from the proband's asymptomatic 72-year-old mother. Other tissues from the mother contained the mutated allele; it was present in 0%-100% of different samples of hair cells and in about 40% of cells from the oral epithelium. Therefore, the mother was a mosaic for the mutation. Since the mutated allele was present in cells derived from all three germ layers, the results indicated that the mutation arose by the late blastocyst stage of development. The results also indicate that assays of blood leukocytes do not always reveal mosaicism or predict phenotypic involvement of tissues, such as blood vessels, that are derived from the same embryonic cells as are leukocytes. 66 refs., 6 figs., 1 tab.

  20. Bartonella henselae trimeric autotransporter adhesin BadA expression interferes with effector translocation by the VirB/D4 type IV secretion system.

    Science.gov (United States)

    Lu, Yun-Yueh; Franz, Bettina; Truttmann, Matthias C; Riess, Tanja; Gay-Fraret, Jérémie; Faustmann, Marco; Kempf, Volkhard A J; Dehio, Christoph

    2013-05-01

    The Gram-negative, zoonotic pathogen Bartonella henselae is the aetiological agent of cat scratch disease, bacillary angiomatosis and peliosis hepatis in humans. Two pathogenicity factors of B. henselae - each displaying multiple functions in host cell interaction - have been characterized in greater detail: the trimeric autotransporter Bartonella adhesin A (BadA) and the type IV secretion system VirB/D4 (VirB/D4 T4SS). BadA mediates, e.g. binding to fibronectin (Fn), adherence to endothelial cells (ECs) and secretion of vascular endothelial growth factor (VEGF). VirB/D4 translocates several Bartonella effector proteins (Beps) into the cytoplasm of infected ECs, resulting, e.g. in uptake of bacterial aggregates via the invasome structure, inhibition of apoptosis and activation of a proangiogenic phenotype. Despite this knowledge of the individual activities of BadA or VirB/D4 it is unknown whether these major virulence factors affect each other in their specific activities. In this study, expression and function of BadA and VirB/D4 were analysed in a variety of clinical B. henselae isolates. Data revealed that most isolates have lost expression of either BadA or VirB/D4 during in vitro passages. However, the phenotypic effects of coexpression of both virulence factors was studied in one clinical isolate that was found to stably coexpress BadA and VirB/D4, as well as by ectopic expression of BadA in a strain expressing VirB/D4 but not BadA. BadA, which forms a dense layer on the bacterial surface, negatively affected VirB/D4-dependent Bep translocation and invasome formation by likely preventing close contact between the bacterial cell envelope and the host cell membrane. In contrast, BadA-dependent Fn binding, adhesion to ECs and VEGF secretion were not affected by a functional VirB/D4 T4SS. The obtained data imply that the essential virulence factors BadA and VirB/D4 are likely differentially expressed during different stages of the infection cycle of

  1. Raman spectra of Cu{sub 2}B{sup II}C{sup IV}X{sub 4}{sup VI} magnetic quaternary semiconductor compounds with tetragonal stannite type structure

    Energy Technology Data Exchange (ETDEWEB)

    Rincón, C., E-mail: crincon@ula.ve; Quintero, M.; Power, Ch.; Moreno, E.; Quintero, E.; Morocoima, M. [Centro de Estudios de Semiconductores, Departamento de Física, Facultad de Ciencias, Universidad de Los Andes, Mérida (Venezuela, Bolivarian Republic of); Henao, J. A.; Macías, M. A. [Grupo de Investigación en Química Estructural, Facultad de Ciencias, Escuela de Química, Universidad Industrial de Santander, Apartado Aéreo 678, Bucaramanga (Colombia)

    2015-05-28

    A comparative study of the Raman spectra of Cu{sub 2}B{sup II}C{sup IV}S{sub 4}{sup VI} and Cu{sub 2}B{sup II}C{sup IV}Se{sub 4}{sup VI}(where B = Mn or Fe) magnetic quaternary semiconductor compounds with stannite-type structure (I4{sup ¯}2m) has been done. Most of the fourteen Raman lines expected for these materials were observed in the spectra. The two strongest lines observed have been assigned to the IR inactive A{sub 1}{sup 1} and A{sub 1}{sup 2} stannite modes that originated from the motion of the S or Se anion around the Cu and C{sup IV} cations remaining at rest. The shift in the frequency of these two lines of about 150 cm{sup −1} to lower energies observed in Cu{sub 2}B{sup II}C{sup IV}Se{sub 4}{sup VI} compounds as compared to those in Cu{sub 2}B{sup II}C{sup IV}S{sub 4}{sup VI} ones, can then be explained as due to the anion mass effect. Based on the fact that values of these frequencies depend mainly on anion mass and bond-stretching forces between nearest-neighbor atoms, the vibrational frequencies v{sup ¯}(A{sub 1}{sup 2}) and v{sup ¯}(A{sub 1}{sup 2}) of both modes for several Cu{sub 2}B{sup II}C{sup IV}X{sub 4}{sup VI} stannite compounds (where X = S, Se, or Te) very close to the experimental data reported for these materials were calculated from a simple model that relates these stretching forces to the anion-cation bond-distances.

  2. Revealing, identifying and assessing ''non-predictable flaws'', crack type IV, by quantitative acoustic emission non-destructive inspection technology, photo-elastic and metallurgical methods

    Energy Technology Data Exchange (ETDEWEB)

    Muravin, G.; Mizrahi, I. [Margan Physical Diagnostics Ltd., Netanya (Israel); Frage, N. [Ben-Gurion Univ. of the Negev, Beer-Sheva (Israel)

    2010-11-15

    QAE NDI technology has been adapted to revealing, identifying and assessing one type of previously non-predictable failure of high energy equipment and piping, known as crack type IV. The research established the following: Cracks of type IV correspond to transverse shear crack mode II, according to the classification used in fracture mechanics and the physics of solids. The length of crack mode II jumps, the velocity of it propagation are significantly higher than in case crack mode I under the same loads. The earliest quantitative statistical Acoustic Emission (AE) indications and peculiarities necessary and sufficient to reveal cracks in specimens loaded by tension or shear loads, especially the differences in the AE power, median Energy, Average Frequency and Hit Rate of the AE signals flow. etc. Based on the findings described above have been formulated requirements and technological solutions for revealing, typifying and assessing crack IV. (Copyright copyright 2010 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

  3. A common variant of staphylococcal cassette chromosome mec type IVa in isolates from Copenhagen, Denmark, is not detected by the BD GeneOhm methicillin-resistant Staphylococcus aureus assay

    DEFF Research Database (Denmark)

    Bartels, Mette Damkjaer; Boye, Kit; Rohde, Susanne Mie

    2009-01-01

    -susceptible Staphylococcus aureus isolates were included as negative controls. Forty-four MRSA isolates were undetectable; of these, 95% harbored SCCmec type IVa, and these included the most-common clone in Copenhagen, spa t024-sequence type 8-IVa. The false-negative MRSA isolates were tested with new primers (analyte...... Copenhagen, Denmark, but also including international isolates, e.g., USA100-1100. Pure cultures of 349 MRSA isolates representing variants of staphylococcal cassette chromosome mec (SCCmec) types I to V and 103 different staphylococcal protein A (spa) types were tested. In addition, 53 methicillin...

  4. Characterization of the Staphylococcus aureus rRNA Methyltransferase Encoded by orfX, the Gene Containing the Staphylococcal Chromosome Cassette mec (SCCmec) Insertion Site*

    Science.gov (United States)

    Boundy, Sam; Safo, Martin K.; Wang, Lei; Musayev, Faik N.; O'Farrell, Heather C.; Rife, Jason P.; Archer, Gordon L.

    2013-01-01

    The gene orfX is conserved among all staphylococci, and its complete sequence is maintained upon insertion of the staphylococcal chromosome cassette mec (SCCmec) genomic island, containing the gene encoding resistance to β-lactam antibiotics (mecA), into its C terminus. The function of OrfX has not been determined. We show that OrfX was constitutively produced during growth, that orfX could be inactivated without altering bacterial growth, and that insertion of SCCmec did not alter gene expression. We solved the crystal structure of OrfX at 1.7 Å and found that it belongs to the S-adenosyl-l-methionine (AdoMet)-dependent α/β-knot superfamily of SPOUT methyltransferases (MTases), with a high structural homology to YbeA, the gene product of the Escherichia coli 70 S ribosomal MTase RlmH. MTase activity was confirmed by demonstrating the OrfX-dependent methylation of the Staphylococcus aureus 70 S ribosome. When OrfX was crystallized in the presence of its AdoMet substrate, we found that each monomer of the homodimeric structure bound AdoMet in its active site. Solution studies using isothermal titration calorimetry confirmed that each monomer bound AdoMet but with different binding affinities (Kd = 52 ± 0.4 and 606 ± 2 μm). In addition, the structure shows that the AdoMet-binding pocket, formed by a deep trefoil knot, contains a bound phosphate molecule, which is the likely nucleotide methylation site. This study represents the first characterization of a staphylococcal ribosomal MTase and provides the first crystal structure of a member of the α/β-knot superfamily of SPOUT MTases in the RlmH or COG1576 family with bound AdoMet. PMID:23150671

  5. The KCNH2-IVS9-28A/G mutation causes aberrant isoform expression and hERG trafficking defect in cardiomyocytes derived from patients affected by Long QT Syndrome type 2.

    Science.gov (United States)

    Mura, Manuela; Mehta, Ashish; Ramachandra, Chrishan J; Zappatore, Rita; Pisano, Federica; Ciuffreda, Maria Chiara; Barbaccia, Vincenzo; Crotti, Lia; Schwartz, Peter J; Shim, Winston; Gnecchi, Massimiliano

    2017-08-01

    Long QT Syndrome type 2 (LQT2) is caused by mutations in the KCNH2 gene that encodes for the α-subunit (hERG) of the ion channel conducting the rapid delayed rectifier potassium current (IKr). We have previously identified a disease causing mutation (IVS9-28A/G) in the branch point of the splicing of KCNH2 intron 9. However, the mechanism through which this mutation causes the disease is unknown. We generated human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) from fibroblasts of two IVS9-28A/G mutation carriers. IVS9-28A/G iPSC-CMs showed prolonged repolarization time, mimicking what observed at the ECG level in the same patients. The expression of the full-length ERG1a isoform resulted reduced, whereas the C-terminally truncated ERG1aUSO isoform was upregulated in mutant iPSC-CMs, with consequent alteration of the physiological ERG1aUSO/ERG1a ratio. Importantly, we observed an impairment of hERG trafficking to the cell membrane. The severity of the alterations in hERG expression and trafficking correlated with the clinical severity of the disease in the two patients under study. Finally, we were able to revert the trafficking defect and reduce the repolarization duration in LQT2 iPSC-CMs using the proteasome inhibitor ALLN. Our results highlight the key role of the KCNH2 intron 9 branch point in the regulation of KCNH2 isoform expression and hERG channel function, and allow to categorize the IVS9-28A/G mutation as LQT2 class 2 mutation. These findings may result in a more personalized clinical management of IVS9-28A/G mutation carriers. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Múltiplos carcinomas basocelulares na região pubiana em uma paciente fototipo IV: relato de caso Multiple basal cell carcinomas in the pubic area in a patient with skin type IV: case report

    Directory of Open Access Journals (Sweden)

    Cristiani Banhos Ferreira

    2011-06-01

    Full Text Available O carcinoma basocelular é a neoplasia maligna cutânea mais comum em humanos, localizando-se, frequentemente, em áreas expostas e em indivíduos de pele clara. Relata-se o caso de uma paciente de 62 anos, faiodérmica, com múltiplas lesões de bordas discretamente elevadas, eritemato-acastanhadas na região pubiana, cujo diagnóstico clínico foi carcinoma basocelular pigmentado, confirmado através do estudo histopatológico. A imunoistoquímica das lesões foi negativa para a pesquisa de papiloma vírusBasal cell carcinoma is the most common type of malignant cutaneous neoplasm in humans, being more frequently located in exposed areas and in fair-skinned individuals. It is reported the case of a 62-yearold female patient, brown-skinned ,with multiple lesions with edges slightly raised, reddish-brown in the pubic region, whose clinical diagnosis was pigmented basal cell carcinoma, confirmed by histopathology. Immunohistochemistry of the lesions was negative for the detection of papilloma virus

  7. Antimicrobial susceptibility and molecular characteristics of methicillin-resistant Staphylococcus aureus in a Japanese secondary care facility.

    Science.gov (United States)

    Kimura, Yumiko; Morinaga, Yoshitomo; Akamatsu, Norihiko; Matsuda, Junichi; Yamaryo, Takeshi; Kawakami, Kenji; Matsuo, Hidenori; Kosai, Kosuke; Uno, Naoki; Hasegawa, Hiroo; Yanagihara, Katsunori

    2016-01-01

    Methicillin-resistant Staphylococcus aureus (MRSA) is prevalent in Japan, and the Staphylococcus cassette chromosome mec (SCCmec) type II is common among hospital-acquired MRSA isolates. Information pertaining to MRSA characteristics is limited, including SCCmec types, in primary or secondary care facilities. A total of 128 MRSA isolates (90 skin and soft tissue isolates and 38 blood isolates) were collected at a secondary care facility, Kawatana Medical Center, from 2005 to 2011. Antimicrobial susceptibility testing for anti-MRSA antibiotics and molecular testing for SCCmec and virulence genes (tst, sec, etb, lukS/F-PV) were performed. Strains positive for lukS/F-PV were analyzed by multilocus sequence typing and phage open-reading frame typing. SCCmec typing in skin and soft tissue isolates revealed that 65.6% had type IV, 22.2% had type II, 8.9% had type I, and 3.3% had type III. In blood isolates, 50.0% had type IV, 47.4% had type II, and 2.6% had type III. Minimum inhibitory concentrations, MIC(50)/MIC(90), against vancomycin, teicoplanin, linezolid, and arbekacin increased slightly in SCCmec II isolates from skin and soft tissue. MICs against daptomycin were similar between sites of isolation. SCCmec type II isolates possess tst and sec genes at a greater frequently than SCCmec type IV isolates. Four lukS/F-PV-positive isolates were divided into two clonal patterns and USA300 was not included. In conclusion, SCCmec type IV was dominant in blood, skin, and soft tissue isolates in a secondary care facility in Japan. Because antimicrobial susceptibility varies with the SCCmec type, SCCmec typing of clinical isolates should be monitored in primary or secondary care facilities. Copyright © 2015 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  8. Early Diagnosis and Treatment of Coronary Heart Disease in Asymptomatic Subjects With Advanced Vascular Atherosclerosis of the Carotid Artery (Type III and IV b Findings Using Ultrasound) and Risk Factors.

    Science.gov (United States)

    Adams, Ansgar; Bojara, Waldemar; Schunk, Klaus

    2018-02-01

    A study was conducted as to whether the early diagnosis of coronary heart disease in asymptomatic subjects with advanced atherosclerosis of the carotid artery which additionally shows at least one risk factor is successful using ultrasound technology. Within the scope of an occupational screening program using subjects from diverse employment sectors, people were given the opportunity to determine their risk of heart attack. During the study the total plaque area (TPA), the maximum plaque thickness in the carotid artery and the PROCAM-Scores of 3,748 healthy men and 2,260 healthy women between the ages of 20 and 64 years were determined. During the subsequent follow-up study 94 subjects sickened. An ultrasound examination of the carotid artery of 79 patients revealed a type III or IV b finding. In a pilot study 33 asymptomatic subjects with a type III or IV b finding in the ultrasound examination were assessed using a computed tomography (CT) coronary angiogram. Additional 10 asymptomatic subjects were examined independently to undergo further cardiac examinations. In the final analysis only five patients had entirely smooth coronary arteries, six had coronary sclerosis, eight had a 30% stenosis, one had a 30-50% stenosis and 23 patients had a stenosis ≥ 50%; and in extreme case, a left main coronary artery stenosis with three-vessel disease. Asymptomatic subjects with advanced atherosclerosis of the carotid artery (type III and type IV b findings) had a high risk for coronary heart disease (CHD). Early treatment of the disease improves the patient's prognosis. A screening consisting in the combination of TPA measurement and determining the maximum plaque thickness is recommended.

  9. Crystallization and preliminary X-ray analysis of the type IV restriction endonuclease ScoMcrA from Streptomyces coelicolor, which cleaves both Dcm-methylated DNA and phosphorothioated DNA.

    Science.gov (United States)

    Liu, Guang; Zhang, Zhenyi; Zhao, Gong; Deng, Zixin; Wu, Geng; He, Xinyi

    2015-01-01

    ScoMcrA is a type IV modification-dependent restriction endonuclease found in the model strain Streptomyces coelicolor. Unlike type I, II and III restriction endonucleases, which cleave unmodified DNA, type IV restriction endonucleases cleave modified DNA, including methylated, hydroxymethylated, glucosyl-hydroxymethylated and phosphorothioated DNA. ScoMcrA targets both Dcm-methylated DNA and phosphorothioated DNA, and makes double-strand breaks 16-28 nt away from the modified nucleotides or the phosphorothioate links. However, the mechanism by which ScoMcrA recognizes these two entirely different types of modification remains unclear. In this study, the ScoMcrA protein was overexpressed, purified and crystallized. The crystals diffracted to 3.35 Å resolution and belonged to space group P2(1)2(1)2(1). The unit-cell parameters were determined to be a=130.19, b=139.36, c=281.01 Å, α=β=γ=90°. These results will facilitate the detailed structural analysis of ScoMcrA and further elucidation of its biochemical mechanism.

  10. Toward the establishment of standardized in vitro tests for lipid-based formulations, part 3: understanding supersaturation versus precipitation potential during the in vitro digestion of type I, II, IIIA, IIIB and IV lipid-based formulations.

    Science.gov (United States)

    Williams, Hywel D; Sassene, Philip; Kleberg, Karen; Calderone, Marilyn; Igonin, Annabel; Jule, Eduardo; Vertommen, Jan; Blundell, Ross; Benameur, Hassan; Müllertz, Anette; Pouton, Colin W; Porter, Christopher J H

    2013-12-01

    Recent studies have shown that digestion of lipid-based formulations (LBFs) can stimulate both supersaturation and precipitation. The current study has evaluated the drug, formulation and dose-dependence of the supersaturation - precipitation balance for a range of LBFs. Type I, II, IIIA/B LBFs containing medium-chain (MC) or long-chain (LC) lipids, and lipid-free Type IV LBF incorporating different doses of fenofibrate or tolfenamic acid were digested in vitro in a simulated intestinal medium. The degree of supersaturation was assessed through comparison of drug concentrations in aqueous digestion phases (APDIGEST) during LBF digestion and the equilibrium drug solubility in the same phases. Increasing fenofibrate or tolfenamic acid drug loads (i.e., dose) had negligible effects on LC LBF performance during digestion, but promoted drug crystallization (confirmed by XRPD) from MC and Type IV LBF. Drug crystallization was only evident in instances when the calculated maximum supersaturation ratio (SR(M)) was >3. This threshold SR(M) value was remarkably consistent across all LBF and was also consistent with previous studies with danazol. The maximum supersaturation ratio (SR(M)) provides an indication of the supersaturation 'pressure' exerted by formulation digestion and is strongly predictive of the likelihood of drug precipitation in vitro. This may also prove effective in discriminating the in vivo performance of LBFs.

  11. Does Type of Tumor Histology Impact Survival among Patients with Stage IIIB/IV Non-Small Cell Lung Cancer Treated with First-Line Doublet Chemotherapy?

    Science.gov (United States)

    Clements, Karen M.; Peltz, Gerson; Faries, Douglas E.; Lang, Kathleen; Nyambose, Joshua; Earle, Craig C.; Sugarman, Katherine P.; Taylor, Douglas C. A.; Thompson, David; Marciniak, Martin D.

    2010-01-01

    Chemotherapy regimens may have differential efficacy by histology in nonsmall cell lung cancer (NSCLC). We examined the impact of histology on survival of patients (N = 2,644) with stage IIIB/IV NSCLC who received first-line cisplatin/carboplatin plus gemcitabine (C/C+G) and cisplatin/carboplatin plus a taxane (C/C+T) identified retrospectively in the SEER cancer registry (1997–2002). Patients with squamous and nonsquamous cell carcinoma survived 8.5 months and 8.1 months, respectively (P = .018). No statistically significant difference was observed in survival between C/C+G and C/C+T in both histologies. Adjusting for clinical and demographic characteristics, the effect of treatment regimen on survival did not differ by histology (P for interaction = .257). There was no statistically significant difference in hazard of death by histology in both groups. These results contrast the predictive role of histology and improved survival outcomes observed for cisplatin-pemetrexed regimens in advanced nonsquamous NSCLC. PMID:22482053

  12. Does Type of Tumor Histology Impact Survival among Patients with Stage IIIB/IV Non-Small Cell Lung Cancer Treated with First-Line Doublet Chemotherapy?

    Directory of Open Access Journals (Sweden)

    Karen M. Clements

    2010-01-01

    Full Text Available Chemotherapy regimens may have differential efficacy by histology in nonsmall cell lung cancer (NSCLC. We examined the impact of histology on survival of patients (N=2,644 with stage IIIB/IV NSCLC who received first-line cisplatin/carboplatin plus gemcitabine (C/C+G and cisplatin/carboplatin plus a taxane (C/C+T identified retrospectively in the SEER cancer registry (1997–2002. Patients with squamous and nonsquamous cell carcinoma survived 8.5 months and 8.1 months, respectively (P=.018. No statistically significant difference was observed in survival between C/C+G and C/C+T in both histologies. Adjusting for clinical and demographic characteristics, the effect of treatment regimen on survival did not differ by histology (P for interaction =.257. There was no statistically significant difference in hazard of death by histology in both groups. These results contrast the predictive role of histology and improved survival outcomes observed for cisplatin-pemetrexed regimens in advanced nonsquamous NSCLC.

  13. Legionella pneumophila Type IV Effectors YlfA and YlfB Are SNARE-Like Proteins that Form Homo- and Heteromeric Complexes and Enhance the Efficiency of Vacuole Remodeling.

    Science.gov (United States)

    Campodonico, Eva M; Roy, Craig R; Ninio, Shira

    2016-01-01

    Legionella pneumophila is a Gram-negative bacterium that can colonize both freshwater protozoa and human alveolar macrophages, the latter infection resulting in Legionnaires' disease. The intracellular lifecycle of L. pneumophila requires extensive manipulation of its host cell, which is carried out by effector proteins that are translocated into the host cell through the Dot/Icm type IV secretion system. This study focuses on a pair of highly similar type IV substrates called YlfA/LegC7 and YlfB/LegC2 that were initially identified in a screen for proteins that cause growth inhibition in yeast. Analysis of truncation mutants revealed that the hydrophobic residues in the Ylf amino termini were required for localization of each protein to the membranes of host cells. Central and carboxy terminal coiled coil domains were found to mediate binding of YlfA and YlfB to themselves and to each other. In vivo, a ΔylfA ΔylfB double mutant strain of L. pneumophila was shown to be defective in establishing a vacuole that supports bacterial replication. This phenotype was subsequently correlated with a decrease in the association of endoplasmic reticulum (ER)-derived vesicles with vacuoles containing ΔylfA ΔylfB mutant bacteria. These data suggest that the Ylf proteins are membrane-associated effectors that enhance remodeling of the L. pneumophila -containing vacuole by promoting association and possibly fusion of ER-derived membrane vesicles with the bacterial compartment.

  14. Legionella pneumophila Type IV Effectors YlfA and YlfB Are SNARE-Like Proteins that Form Homo- and Heteromeric Complexes and Enhance the Efficiency of Vacuole Remodeling.

    Directory of Open Access Journals (Sweden)

    Eva M Campodonico

    Full Text Available Legionella pneumophila is a Gram-negative bacterium that can colonize both freshwater protozoa and human alveolar macrophages, the latter infection resulting in Legionnaires' disease. The intracellular lifecycle of L. pneumophila requires extensive manipulation of its host cell, which is carried out by effector proteins that are translocated into the host cell through the Dot/Icm type IV secretion system. This study focuses on a pair of highly similar type IV substrates called YlfA/LegC7 and YlfB/LegC2 that were initially identified in a screen for proteins that cause growth inhibition in yeast. Analysis of truncation mutants revealed that the hydrophobic residues in the Ylf amino termini were required for localization of each protein to the membranes of host cells. Central and carboxy terminal coiled coil domains were found to mediate binding of YlfA and YlfB to themselves and to each other. In vivo, a ΔylfA ΔylfB double mutant strain of L. pneumophila was shown to be defective in establishing a vacuole that supports bacterial replication. This phenotype was subsequently correlated with a decrease in the association of endoplasmic reticulum (ER-derived vesicles with vacuoles containing ΔylfA ΔylfB mutant bacteria. These data suggest that the Ylf proteins are membrane-associated effectors that enhance remodeling of the L. pneumophila -containing vacuole by promoting association and possibly fusion of ER-derived membrane vesicles with the bacterial compartment.

  15. Vancomycin heteroresistant community associated methicillin-resistant Staphylococcus aureus ST72-SCCmecIVa strain colonizing the nostrils of a five-year-old Spanish girl.

    Science.gov (United States)

    Varona-Barquín, Aketza; Iglesias-Losada, Juan José; Ezpeleta, Guillermo; Eraso, Elena; Quindós, Guillermo

    2017-03-01

    During a community methicillin-resistant Staphylococcus aureus (MRSA) nasal colonization study, an MRSA strain with vancomycin hetero-resistance (h-VISA) was isolated from a five year-old girl with tetralogy of Fallot without previous exposure to vancomycin. An extended nasal colonization study was performed on all her close relatives. Only the patient and her sister were colonized by an h-VISA MRSA strain (clone USA 700, ST72, t148, agr 1 and SCCmec IVa). Mupirocin decolonisation was effective in the elder sister. A new nasal decolonisation in the younger girl using fusidic acid was also successful. However, after decolonisation both sisters were colonized by a methicillin-susceptible S. aureus (ST30, t012 and agr 3) previously isolated from their mother's nostrils. As S. aureus have a great capacity to spread among people in close contact, knowledge of a patients' colonization status, tracing contacts, and a correct management are critical issues for the successful containment of multiresistant staphylococci. Copyright © 2016 Elsevier España, S.L.U. and Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.

  16. Reductive cleavage and reformation of the interchain and intrachain disulfide bonds in the globular hexameric domain NC1 involved in network assembly of basement membrane collagen (type IV).

    Science.gov (United States)

    Weber, S; Dölz, R; Timpl, R; Fessler, J H; Engel, J

    1988-08-01

    The formation of collagen IV dimers in the extracellular space requires the association of two C-terminal globular domains giving rise to a large hexameric structure NC1 (Mr = 170,000). NC1 hexamer was purified from collagenase digests of a mouse tumor and several human tissues. It was shown by electrophoresis to consist of two kinds of cross-linked, dimeric segments, Da and Db (Mr about 50,000), and monomeric segments in a molar ratio of about 3:1. In the native hexamers free SH groups were detectable by N-[14C]ethylmaleimide and other sulfhydryl reagents. They account for 4-11% of the total number of cysteine residues with some variations between preparations from different sources and in the distribution between monomers and dimers. Reduction with 10 mM dithioerythritol under non-denaturing condition completely converted dimers into monomers and allowed the alkylation of all twelve cysteine residues present in each monomeric NC1 segment. A monomeric intermediate with four to six free SH groups and a higher electrophoretic mobility than the final product was observed. Generation of this intermediate from dimers Da and Db follows apparently different routes proceeding either directly or through a dimeric intermediate respectively. The time course of conversion is best described by a mechanism consisting of two (Db) or three (Da) consecutive steps with pseudo-first-order rate constants ranging from 0.14 ms-1 to 0.5 ms-1. Glutathione-catalyzed reoxidation of completely reduced NC1 in the presence of 2 M urea results in a product indistinguishable from native material by ultracentrifugation and electrophoresis pattern. The data suggest that in situ formation of NC1 structures is catalyzed by a small fraction (5-10%) of intrinsic SH groups leading to the formation and stabilization of dimers by rearrangement of disulfide bonds.

  17. IVS1 −397T>C Estrogen Receptor α Polymorphism Is Associated with Low-Grade Systemic Inflammatory Response in Type 1 Diabetic Girls

    Directory of Open Access Journals (Sweden)

    Monika Ryba-Stanisławowska

    2014-01-01

    Full Text Available Purpose. The study aimed to investigate the influence of estrogen receptor α (ER-α genotypes on inflammatory response and development of microvascular complications in girls with type 1 diabetes. Methods. 152 young regularly menstruating girls with diagnosed type 1 diabetes and 84 young, healthy menstruating girls were recruited. ER-α genotyping was carried out by PCR. Serum concentrations of 17β-estradiol, as well as IL-6, TNF-α, VEGF, and IL-10, were measured. CD4+Foxp3+ TH17 cells were isolated and analyzed by flow cytometry. Results. Type 1 diabetic girls carrying TT genotype were characterized by the lowest serum estradiol level and IL-10 and highest IL-6, TNF-α , and VEGF. The association between the level of certain cytokine and the genetic variant of estrogen receptor α polymorphism was analyzed. Frequencies of CD4+Foxp3+ TH17 cells were also enhanced in TT bearing girls with type 1 diabetes and correlated with the level of analyzed cytokines. In addition, the correlation between serum estradiol level and cytokine concentrations was observed. Conclusions. We propose that TT variant of estrogen receptor α polymorphism may be associated with enhanced inflammatory response, which in turn may lead to acceleration of diabetic retino- and nephropathy in girls with type 1 diabetes. This finding may help the physicians to predict the onset and progression of diabetic microvascular complications.

  18. SDSS-IV MaNGA: Probing the Kinematic Morphology–Density Relation of Early-type Galaxies with MaNGA

    Science.gov (United States)

    Greene, J. E.; Leauthaud, A.; Emsellem, E.; Goddard, D.; Ge, J.; Andrews, B. H.; Brinkman, J.; Brownstein, J. R.; Greco, J.; Law, D.; Lin, Y.-T.; Masters, K. L.; Merrifield, M.; More, S.; Okabe, N.; Schneider, D. P.; Thomas, D.; Wake, D. A.; Yan, R.; Drory, N.

    2017-12-01

    The “kinematic” morphology–density relation for early-type galaxies posits that those galaxies with low angular momentum are preferentially found in the highest-density regions of the universe. We use a large sample of galaxy groups with halo masses {10}12.5MaNGA) survey to examine whether there is a correlation between local environment and rotational support that is independent of stellar mass. We find no compelling evidence for a relationship between the angular momentum content of early-type galaxies and either local overdensity or radial position within the group at fixed stellar mass.

  19. Management of a Type I Hypersensitivity Reaction to IV Etoposide in a Woman with a Yolk Sac Tumor: A Case Report

    Directory of Open Access Journals (Sweden)

    David Starks

    2011-01-01

    Full Text Available Type I hypersensitivity reactions to intravenous administration of etoposide are extremely rare. Etoposide is an essential component of several chemotherapy regimens used in gynecologic oncology, and discontinuation of this drug during a course of treatment should only be due to severe patient intolerance. We report the successful use of intravenous etoposide phosphate as a substitute drug in a patient with a yolk sac tumor who manifested a Type I hypersensitivity to intravenous etoposide. The patient ultimately completed all 4 cycles of bleomycin, etoposide, cisplatin (BEP using etoposide phosphate as a substitute drug.

  20. A simplified multiplex PCR assay for fast and easy discrimination of globally distributed staphylococcal cassette chromosome mec types in meticillin-resistant Staphylococcus aureus

    NARCIS (Netherlands)

    E. Ghaznavi Rad (Ehsanollah); N.S. Mariana (Nor Shamsudin); Z. Sekawi (Zamberi); A.F. van Belkum (Alex); V. Neela (Vasanthakumari)

    2010-01-01

    textabstractA multiplex PCR assay was developed for the identification of major types and subtypes of staphylococcal cassette chromosome mec (SCCmec) in meticillin-resistant Staphylococcus aureus (MRSA) strains. The method uses a novel 9 valent multiplex PCR plus two primer pairs for S. aureus

  1. Alternative Agents in Type 1 Diabetes in Addition to Insulin Therapy: Metformin, Alpha-Glucosidase Inhibitors, Pioglitazone, GLP-1 Agonists, DPP-IV Inhibitors, and SGLT-2 Inhibitors.

    Science.gov (United States)

    DeGeeter, Michelle; Williamson, Bobbie

    2016-04-01

    Insulin is the mainstay of current treatment for patients with type 1 diabetes mellitus (T1DM). Due to increasing insulin resistance, insulin doses are often continually increased, which may result in weight gain for patients. Medications currently approved for the treatment of type 2 diabetes offer varying mechanisms of action that can help to reduce insulin resistance and prevent or deter weight gain. A MEDLINE search was conducted to review literature evaluating the use of metformin, alpha-glucosidase inhibitors, pioglitazone, glucagon-like peptide 1 agonists, dipeptidyl peptidase, and sodium-dependent glucose transporter 2 inhibitors, in patients with T1DM. Varying results were found with some benefits including reductions in hemoglobin A1c, decreased insulin doses, and favorable effects on weight. Of significance, a common fear of utilizing multiple therapies for diabetes treatment is the risk of hypoglycemia, and this review displayed limited evidence of hypoglycemia with multiple agents. © The Author(s) 2014.

  2. Coxiella burnetii Employs the Dot/Icm Type IV Secretion System to Modulate Host NF-κB/RelA Activation.

    Science.gov (United States)

    Mahapatra, Saugata; Gallaher, Brandi; Smith, Sydni Caet; Graham, Joseph G; Voth, Daniel E; Shaw, Edward I

    2016-01-01

    Coxiella burnetii is the causative agent of Q fever and an obligate intracellular pathogen in nature that survives and grows in a parasitophorous vacuole (PV) within eukaryotic host cells. C. burnetii promotes intracellular survival by subverting apoptotic and pro-inflammatory signaling pathways that are typically regulated by nuclear transcription factor-κB (NF-κB). We and others have demonstrated that C. burnetii NMII proteins inhibit expression of pro-inflammatory cytokines and induce expression of anti-apoptotic genes during infection. Here, we demonstrate that C. burnetii promotes intracellular survival by modulating NF-κB subunit p65 (RelA) phosphorylation, and thus activation, in a Type Four B Secretion System (T4BSS)-dependent manner. Immunoblot analysis of RelA phosphorylated at serine-536 demonstrated that C. burnetii increases NF-κB activation via the canonical pathway. However, RelA phosphorylation levels were even higher in infected cells where bacterial protein or mRNA synthesis was inhibited. Importantly, we demonstrate that inhibition of RelA phosphorylation impairs PV formation and C. burnetii growth. We found that a T4BSS-defective mutant (CbΔdotA) elicited phosphorylated RelA levels similar to those of wild type C. burnetii infection treated with Chloramphenicol. Moreover, cells infected with CbΔdotA or wild type C. burnetii treated with Chloramphenicol showed similar levels of GFP-RelA nuclear localization, and significantly increased localization compared to wild type C. burnetii infection. These data indicate that without de novo protein synthesis and a functional T4BSS, C. burnetii is unable to modulate NF-κB activation, which is crucial for optimal intracellular growth.

  3. Study of a SiGeSn/GeSn/SiGeSn structure toward direct bandgap type-I quantum well for all group-IV optoelectronics.

    Science.gov (United States)

    Ghetmiri, Seyed Amir; Zhou, Yiyin; Margetis, Joe; Al-Kabi, Sattar; Dou, Wei; Mosleh, Aboozar; Du, Wei; Kuchuk, Andrian; Liu, Jifeng; Sun, Greg; Soref, Richard A; Tolle, John; Naseem, Hameed A; Li, Baohua; Mortazavi, Mansour; Yu, Shui-Qing

    2017-02-01

    A SiGeSn/GeSn/SiGeSn single quantum well structure was grown using an industry standard chemical vapor deposition reactor with low-cost commercially available precursors. The material characterization revealed the precisely controlled material growth process. Temperature-dependent photoluminescence spectra were correlated with band structure calculation for a structure accurately determined by high-resolution x-ray diffraction and transmission electron microscopy. Based on the result, a systematic study of SiGeSn and GeSn bandgap energy separation and barrier heights versus material compositions and strain was conducted, leading to a practical design of a type-I direct bandgap quantum well.

  4. Impact of baseline BMI on glycemic control and weight change with metformin monotherapy in Chinese type 2 diabetes patients: phase IV open-label trial.

    Directory of Open Access Journals (Sweden)

    Linong Ji

    Full Text Available Differences exist between treatment recommendations regarding the choice of metformin as first-line therapy for type 2 diabetes patients according to body mass index (BMI. This study compared the efficacy of metformin monotherapy among normal-weight, overweight, and obese patients with newly diagnosed type 2 diabetes.In this prospective, multicenter, open-label study in China, patients aged 23-77 years were enrolled 1∶1:1 according to baseline BMI: normal-weight (BMI 18.5-23.9 kg/m(2; n = 125; overweight (BMI 24.0-27.9 kg/m(2; n = 122 or obese (BMI ≥28 kg/m(2; n = 124. Extended-release metformin was administered for 16 weeks (500 mg/day, up-titrated weekly to a maximum 2,000 mg/day. The primary efficacy endpoint was the effect of baseline BMI on glycemic control with metformin monotherapy, measured as the change from baseline in glycosylated hemoglobin (HbA1c at week 16 compared among BMI groups using ANCOVA. Other endpoints included comparisons of metformin's effects on fasting plasma glucose (FPG, lipid levels and body weight.Mean HbA1c decreases at week 16, adjusted for baseline values, were -1.84%, -1.78% and -1.78% in normal-weight, overweight and obese patients, (P = 0.664; body weight decreased by 2.4%, 3.9% and 3.5%, respectively. FPG levels decreased similarly over time in all BMI groups (P = 0.461 and changes from baseline in high-density lipoprotein cholesterol (HDL-C and low-density lipoprotein cholesterol (LDL-C did not differ significantly among BMI groups at week 16 (P = 0.143 and 0.451, respectively.Baseline BMI had no impact on glycemic control, weight change or other efficacy measures with metformin monotherapy. These data suggest that normal-weight type 2 diabetes patients would derive the same benefits from first-line treatment with metformin as overweight and obese patients, and are not at increased risk of excess weight loss.ClinicalTrials.gov NCT00778622.

  5. Cementless total hip arthroplasty for patients with Crowe type III or IV developmental dysplasia of the hip: two-stage total hip arthroplasty following skeletal traction after soft tissue release for irreducible hips.

    Science.gov (United States)

    Yoon, Pil Whan; Kim, Jung Il; Kim, Dong Ok; Yu, Cheol Hwan; Yoo, Jeong Joon; Kim, Hee Joong; Yoon, Kang Sup

    2013-09-01

    Total hip arthroplasty (THA) for severe developmental dysplasia of the hip (DDH) is a technically demanding procedure for arthroplasty surgeons, and it is often difficult to reduce the hip joint without soft tissue release due to severe flexion contracture. We performed two-stage THAs in irreducible hips with expected lengthening of the affected limb after THA of over 2.5 cm or with flexion contractures of greater than 30 degrees in order to place the acetabular cup in the true acetabulum and to prevent neurologic deficits associated with acute elongation of the limb. The purpose of this study is to evaluate the outcomes of cementless THA in patients with severe DDH with a special focus on the results of two-stage THA. Retrospective clinical and radiological evaluations were done on 17 patients with Crowe type III or IV developmental DDH treated by THA. There were 14 women and 3 men with a mean age of 52.3 years. Follow-ups averaged 52 months. Six cases were treated with two-stage THA followed by surgical hip liberalization and skeletal traction for 2 weeks. The mean Harris hip score improved from 40.9 to 89.1, and mean leg length discrepancy (LLD) in 13 unilateral cases was reduced from 2.95 to 0.8 cm. In the patients who underwent two-stage surgery, no nerve palsy was observed, and the single one-stage patient with incomplete peroneal nerve palsy recovered fully 4 weeks postoperatively. The short-term clinical and radiographic outcomes of primary cementless THA for patients with Crowe type III or IV DDH were encouraging. Two-stage THA followed by skeletal traction after soft tissue release could provide alternative solutions to the minimization of limb shortenings or LLD without neurologic deficits in highly selected patients.

  6. Different Clonal Complexes of Methicillin-Resistant Staphylococcus aureus Are Disseminated in the Euregio Meuse-Rhine Region

    Science.gov (United States)

    Deurenberg, Ruud H.; Vink, Cornelis; Oudhuis, Guy J.; Mooij, Jascha E.; Driessen, Christel; Coppens, Guy; Craeghs, Jos; De Brauwer, Els; Lemmen, Sebastian; Wagenvoort, Hans; Friedrich, Alexander W.; Scheres, Jacques; Stobberingh, Ellen E.

    2005-01-01

    The Euregio Meuse-Rhine (EMR) is formed by the border regions of Belgium, Germany, and The Netherlands. Cross-border health care requires infection control measures, in particular since the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) differs among the three countries. To investigate the dissemination of MRSA in the EMR, 152 MRSA isolates were characterized by pulsed-field gel electrophoresis (PFGE), SCCmec typing, and multilocus sequence typing. PFGE revealed major clonal groups A, G, L, and Q, suggesting dissemination of MRSA in the EMR. Group A harbored mainly SCCmec type III and sequence types (STs) 239 and 241. The majority of the strains from group G harbored SCCmec type I and ST8 and ST247, whereas most strains from group L carried either SCCmec type IV or type I. Within group L, ST8 and ST228 were found, belonging to clonal complexes 8 and 5, respectively. Most strains from group Q included SCCmec type II and were sequence typed as ST225. Both ST225-MRSA-II and ST241-MRSA-III were novel findings in Germany. In addition, the SCCmec type of two isolates has not been described previously. One strain was classified as SCCmec type III but harbored the pls gene and the dcs region. Another strain was characterized as SCCmec type IV but lacked the dcs region. In addition, one isolate harbored both SCCmec type V and Panton-Valentine leukocidin. Finally, the SCCmec type of the strains was found to be correlated with the antibiotic susceptibility pattern. PMID:16189107

  7. Preliminary trial to investigate temperature of the iPulse intense pulsed light (IPL) glass transmission block during treatment of Fitzpatrick II, IV, V, and VI skin types.

    Science.gov (United States)

    Ash, C; Town, G A; Martin, G R

    2007-03-01

    The glass transmission block, a key component of all intense pulsed light (IPL) devices, is responsible for the delivery of IPL energy from the xenon discharge lamp to hair and skin structures during treatment. The purpose of this study was to investigate the variation in temperature of the quartz glass block used in the iPulse (CyDen, Swansea, UK) handset during typical hair removal treatments of Asian and Afro-Caribbean skin types. Initial results from four subjects indicated that the temperature of the glass transmission block did not exceed 45 degrees C during any of the treatments. Furthermore, the development of the temperature measurement methodology described in this paper will enable the comparison of data from different IPL systems to be undertaken in a subsequent larger scale trial.

  8. Type IV renal tubular acidosis following resolution of acute kidney injury and disseminated intravascular coagulation due to hump-nosed viper bite

    Directory of Open Access Journals (Sweden)

    S Karunarathne

    2013-01-01

    Full Text Available Hump-nosed viper bite can cause acute kidney injury (AKI and disseminated intravascular coagulation. In some patients, it can cause chronic kidney disease necessitating life-long renal replacement therapy. Lack of effective antivenom makes the management of these patients difficult. A 51-year-old Sri Lankan male was admitted with AKI and disseminated intravascular coagulation following a hump-nosed viper bite. He made a complete recovery with blood product support and hemodialysis. Renal biopsy was performed as his renal recovery was prolonged which revealed patchy tubular atrophy and interstitial inflammation suggestive of subacute interstitial nephritis. Later, he presented with hyperkalemic paralysis and acidosis. A diagnosis of late onset type 4 renal tubular acidosis was made and he responded well to a course of fludrocortisone.

  9. Experimental studies of immunologically mediated enteropathy: IV. Correlation between immune effector mechanisms and type of enteropathy during a GvHR in neonatal mice of different ages.

    Science.gov (United States)

    Felstein, M V; Mowat, A M

    1988-01-01

    We have used the intestinal phase of the graft-versus-host-reaction (GvHR) in unirradiated F1 mice as a model for enteropathy due to cell-mediated immunity (CMI). Injection of neonatal (CBA x BALB/c)F1 mice less than 48 h old with CBA spleen cells produced an acute GvHR, which was associated with runting and severe intestinal damage, characterized by villus atrophy. These animals developed specific cytotoxic T lymphocyte (CTL) activity and invariably died. In contrast, 7-day-old F1 mice with GvHR developed a proliferative GvHR, characterized by intense splenomegaly, NK cell activation and intestinal crypt hyperplasia. These mice did not lose weight, had no villus atrophy or CTL activity and all recovered. A similar proliferative phase was also found to precede the established GvHR in 1-2-day-old hosts. Induction of a GvHR in 5-day-old hosts produced a disease with some characteristics of both proliferative and destructive GvHR, with some mice developing weight loss and villus atrophy, while others showed only crypt hyperplasia and NK cell activation. However, there was very little specific CTL activity in any of these animals. These results indicate that markedly different forms of GvHR can be induced in mice during the first week of life and that these are associated with different pathological effects. Although the immunological mechanisms which are activated may also differ between the types of GvHR, our findings support the hypothesis that intestinal damage which includes villus atrophy is merely a progressive form of the delayed type hypersensitivity responsible for a proliferative enteropathy. PMID:3396213

  10. SDSS-IV MaNGA: Global stellar population and gradients for about 2000 early-type and spiral galaxies on the mass-size plane

    Science.gov (United States)

    Li, Hongyu; Mao, Shude; Cappellari, Michele; Ge, Junqiang; Long, R. J.; Li, Ran; Mo, HJ; Li, Cheng; Zheng, Zheng; Bundy, Kevin; Thomas, Daniel; Brownstein, Joel R.; Lopes, Alexandre Roman; Law, David R.; Drory, Niv

    2018-02-01

    We perform full spectrum fitting stellar population analysis and Jeans Anisotropic modelling (JAM) of the stellar kinematics for about 2000 early-type galaxies (ETGs) and spiral galaxies from the MaNGA DR14 sample. Galaxies with different morphologies are found to be located on a remarkably tight mass plane which is close to the prediction of the virial theorem, extending previous results for ETGs. By examining an inclined projection (`the mass-size' plane), we find that spiral and early-type galaxies occupy different regions on the plane, and their stellar population properties (i.e. age, metallicity and stellar mass-to-light ratio) vary systematically along roughly the direction of velocity dispersion, which is a proxy for the bulge fraction. Galaxies with higher velocity dispersions have typically older ages, larger stellar mass-to-light ratios and are more metal rich, which indicates that galaxies increase their bulge fractions as their stellar populations age and become enriched chemically. The age and stellar mass-to-light ratio gradients for low-mass galaxies in our sample tend to be positive (centregalaxies are negative. The metallicity gradients show a clear peak around velocity dispersion log10σe ≈ 2.0, which corresponds to the critical mass ˜3 × 1010M⊙ of the break in the mass-size relation. Spiral galaxies with large mass and size have the steepest gradients, while the most massive ETGs, especially above the critical mass M_crit≳ 2× 10^{11} M_{\\odot}, where slow rotator ETGs start dominating, have much flatter gradients. This may be due to differences in their evolution histories, e.g. mergers.

  11. Inhibition of microRNA-214-5p promotes cell survival and extracellular matrix formation by targeting collagen type IV alpha 1 in osteoblastic MC3T3-E1 cells

    Science.gov (United States)

    Li, Q. S.; Meng, F. Y.; Zhao, Y. H.; Jin, C. L.; Tian, J.

    2017-01-01

    Objectives This study aimed to investigate the functional effects of microRNA (miR)-214-5p on osteoblastic cells, which might provide a potential role of miR-214-5p in bone fracture healing. Methods Blood samples were obtained from patients with hand fracture or intra-articular calcaneal fracture and from healthy controls (HCs). Expression of miR-214-5p was monitored by qRT-PCR at day 7, 14 and 21 post-surgery. Mouse osteoblastic MC3T3-E1 cells were transfected with antisense oligonucleotides (ASO)-miR-214-5p, collagen type IV alpha 1 (COL4A1) vector or their controls; thereafter, cell viability, apoptotic rate, and the expression of collagen type I alpha 1 (COL1A1), type II collagen (COL-II), and type X collagen (COL-X) were determined. Luciferase reporter assay, qRT-PCR, and Western blot were performed to ascertain whether COL4A1 was a target of miR-214-5p. Results Plasma miR-214-5p was highly expressed in patients with bone fracture compared with HCs after fracture (p COL4A1 was a target of miR-214-5p, and was negatively regulated by miR-214-5p (p COL4A1 showed a similar impact on cell viability, apoptotic rate, and COL1A1, COL-II, and COL-X expressions inhibiting miR-214-5p (p COL4A1. Cite this article: Q. S. Li, F. Y. Meng, Y. H. Zhao, C. L. Jin, J. Tian, X. J. Yi. Inhibition of microRNA-214-5p promotes cell survival and extracellular matrix formation by targeting collagen type IV alpha 1 in osteoblastic MC3T3-E1 cells. Bone Joint Res 2017;6:464–471. DOI: 10.1302/2046-3758.68.BJR-2016-0208.R2 PMID:28784704

  12. NNDSS - Table IV. Tuberculosis

    Data.gov (United States)

    U.S. Department of Health & Human Services — NNDSS - Table IV. Tuberculosis - 2014.This Table includes total number of cases reported in the United States, by region and by states, in accordance with the...

  13. NNDSS - Table IV. Tuberculosis

    Data.gov (United States)

    U.S. Department of Health & Human Services — NNDSS - Table IV. Tuberculosis - 2015.This Table includes total number of cases reported in the United States, by region and by states, in accordance with the...

  14. NNDSS - Table IV. Tuberculosis

    Data.gov (United States)

    U.S. Department of Health & Human Services — NNDSS - Table IV. Tuberculosis - 2016.This Table includes total number of cases reported in the United States, by region and by states, in accordance with the...

  15. Neisseria meningitidis differentially controls host cell motility through PilC1 and PilC2 components of type IV Pili.

    Directory of Open Access Journals (Sweden)

    Philippe C Morand

    Full Text Available Neisseria meningitidis is a strictly human pathogen that has two facets since asymptomatic carriage can unpredictably turn into fulminant forms of infection. Meningococcal pathogenesis relies on the ability of the bacteria to break host epithelial or endothelial cellular barriers. Highly restrictive, yet poorly understood, mechanisms allow meningococcal adhesion to cells of only human origin. Adhesion of encapsulated and virulent meningococci to human cells relies on the expression of bacterial type four pili (T4P that trigger intense host cell signalling. Among the components of the meningococcal T4P, the concomitantly expressed PilC1 and PilC2 proteins regulate pili exposure at the bacterial surface, and until now, PilC1 was believed to be specifically responsible for T4P-mediated meningococcal adhesion to human cells. Contrary to previous reports, we show that, like PilC1, the meningococcal PilC2 component is capable of mediating adhesion to human ME180 epithelial cells, with cortical plaque formation and F-actin condensation. However, PilC1 and PilC2 promote different effects on infected cells. Cellular tracking analysis revealed that PilC1-expressing meningococci caused a severe reduction in the motility of infected cells, which was not the case when cells were infected with PilC2-expressing strains. The amount of both total and phosphorylated forms of EGFR was dramatically reduced in cells upon PilC1-mediated infection. In contrast, PilC2-mediated infection did not notably affect the EGFR pathway, and these specificities were shared among unrelated meningococcal strains. These results suggest that meningococci have evolved a highly discriminative tool for differential adhesion in specific microenvironments where different cell types are present. Moreover, the fine-tuning of cellular control through the combined action of two concomitantly expressed, but distinctly regulated, T4P-associated variants of the same molecule (i.e. PilC1 and Pil

  16. A new fructose-free, resistant-starch type IV-enriched enteral formula improves glycaemic control and cardiovascular risk biomarkers when administered for six weeks to elderly diabetic patients.

    Science.gov (United States)

    Mesa García, María Dolores; García-Rodríguez, Cruz Erika; Rico, María de la Cruz; Aguilera, Concepción María; Pérez-Rodríguez, Milagros; Pérez-de-la-Cruz, Antonio Jesús; Gil, Ángel

    2017-02-01

    Reducing the dietary glycaemic response has been proposed as a way to reduce the risk of diabetes complications. The aim of the present study was to evaluate the glycaemic control and cardiovascular risk biomarkers in fragile, elderly type 2 diabetes patients after the intake of a new fructose-free diabetes-specific formula enriched with resistant-starch type IV and high in monounsaturated fatty acids. Forty-one type 2 diabetes patients aged 78.9 ± 2.8 years were fed exclusively with an enteral diabetes-specific formula for 6 weeks. Data were collected at baseline and after 6 weeks of feeding. Carbohydrate and lipid metabolism and inflammatory and cardiovascular risk biomarkers were measured to evaluated the course of diabetes complications. Blood glycated haemoglobin significantly decreased after the intervention (6.1 ± 0.1 vs. 5.8 ± 0.1 %; p< 0,045), as well as monocyte chemotactic protein-1 and soluble E-selectin (p < 0.05), while soluble vascular cell adhesion molecule and plasminogen activator inhibitor-1 tended to decrease from baseline to 6 weeks (p = 0.084 and p = 0.05, respectively). The new product improves glycaemic control and cardiovascular risk without altering lipid metabolism, which is useful for the prevention of diabetic complications. Longer intervention studies are needed in order to validate these results in a larger population.

  17. Traveling-wave solutions of a modified Hodgkin-Huxley type neural model via Novel analytical results for nonlinear transmission lines with arbitrary I(V characteristics

    Directory of Open Access Journals (Sweden)

    Valentino Anthony Simpao

    2000-01-01

    Full Text Available Herein an enhanced Hodgkin-Huxley (H-H type model of neuron dynamics is solved analytically via formal methods. Our model is a variant of an earlier one by M.A. Mahrous and H.Y. Alkahby [1]. Their modified model is realized by a hyperbolic quasi-linear diffusion operator with time-delay parameters; this compared to the original H-H model with standard parabolic quasi-linear diffusion operator and no time-delay parameters. Besides these features, the present model also incorporates terms describing signal dissipation into the background substrate (e.g., conductance to ground, making it more experimentally amenable. The solutions which results via the present scheme are of traveling-wave profile, which agree qualitatively with those observed in actual electro-physiological measurements made on the neural systems originally studied by H-H These results confirm the physiological soundness of the enhanced model and of the preliminary assumptions which motivated the present solution strategy; the comparison of the present results with actual electro-physiological data displays shall appear in later publications.

  18. Livestock-Associated Methicillin Resistant and Methicillin Susceptible Staphylococcus aureus Sequence Type (CC)1 in European Farmed Animals: High Genetic Relatedness of Isolates from Italian Cattle Herds and Humans

    DEFF Research Database (Denmark)

    Alba, Patricia; Feltrin, Fabiola; Cordaro, Gessica

    2015-01-01

    Methicillin-resistant Staphylococcus aureus (MRSA) Sequence Type (ST)1, Clonal Complex( CC) 1, SCCmec V is one of the major Livestock-Associated (LA-) lineages in pig farming industry in Italy and is associated with pigs in other European countries. Recently, it has been increasingly detected in ...

  19. The All-Alpha Domains of Coupling Proteins from the Agrobacterium tumefaciens VirB/VirD4 and Enterococcus faecalis pCF10-Encoded Type IV Secretion Systems Confer Specificity to Binding of Cognate DNA Substrates.

    Science.gov (United States)

    Whitaker, Neal; Chen, Yuqing; Jakubowski, Simon J; Sarkar, Mayukh K; Li, Feng; Christie, Peter J

    2015-07-01

    Bacterial type IV coupling proteins (T4CPs) bind and mediate the delivery of DNA substrates through associated type IV secretion systems (T4SSs). T4CPs consist of a transmembrane domain, a conserved nucleotide-binding domain (NBD), and a sequence-variable helical bundle called the all-alpha domain (AAD). In the T4CP structural prototype, plasmid R388-encoded TrwB, the NBD assembles as a homohexamer resembling RecA and DNA ring helicases, and the AAD, which sits at the channel entrance of the homohexamer, is structurally similar to N-terminal domain 1 of recombinase XerD. Here, we defined the contributions of AADs from the Agrobacterium tumefaciens VirD4 and Enterococcus faecalis PcfC T4CPs to DNA substrate binding. AAD deletions abolished DNA transfer, whereas production of the AAD in otherwise wild-type donor strains diminished the transfer of cognate but not heterologous substrates. Reciprocal swaps of AADs between PcfC and VirD4 abolished the transfer of cognate DNA substrates, although strikingly, the VirD4-AADPcfC chimera (VirD4 with the PcfC AAD) supported the transfer of a mobilizable plasmid. Purified AADs from both T4CPs bound DNA substrates without sequence preference but specifically bound cognate processing proteins required for cleavage at origin-of-transfer sequences. The soluble domains of VirD4 and PcfC lacking their AADs neither exerted negative dominance in vivo nor specifically bound cognate processing proteins in vitro. Our findings support a model in which the T4CP AADs contribute to DNA substrate selection through binding of associated processing proteins. Furthermore, MOBQ plasmids have evolved a docking mechanism that bypasses the AAD substrate discrimination checkpoint, which might account for their capacity to promiscuously transfer through many different T4SSs. For conjugative transfer of mobile DNA elements, members of the VirD4/TraG/TrwB receptor superfamily bind cognate DNA substrates through mechanisms that are largely undefined. Here

  20. Gene flow across genus barriers – Conjugation of Dinoroseobacter shibae´s 191-kb killer plasmid into Phaeobacter inhibens and AHL-mediated expression of type IV secretion systems

    Directory of Open Access Journals (Sweden)

    Diana ePatzelt

    2016-05-01

    Full Text Available Rhodobacteraceae harbor a conspicuous wealth of extrachromosomal replicons (ECRs and therefore the exchange of genetic material via horizontal transfer has been supposed to be a major evolutionary driving force. Many plasmids in the roseobacter group encode type IV secretion systems (T4SS that generally mediate transfer of proteins and/or DNA into host cells, but no experimental evidence of either has yet been provided. Dinoroseobacter shibae contains five ECRs that are crucial for anaerobic growth, survival under starvation and the pathogenicity of this model organism. Here we tagged two syntenous but compatible RepABC-type plasmids of 191 kb and 126-kb size, each harboring a T4SS, with antibiotic resistance genes and demonstrated their conjugational transfer into a distantly related roseo-bacter species, namely Phaeobacter inhibens. Pulsed field gel electrophoresis showed transfer of those replicons into the recipient both individually but also together documenting the efficiency of conjugation. We then studied the influence of externally added quorum sensing (QS signals on the expression of the T4SS located on the sister plasmids. A QS deficient D. shibae null mutant (ΔluxI1 lacking synthesis of N-acyl-homoserine lactones (AHLs was cultivated with a wide spectrum of chemically diverse long-chain AHLs. All AHLs with lengths of the acid side-chain >14 reverted the ΔluxI1 phenotype to wild-type. Expression of the T4SS was induced above wild-type level. We hypothesize that conjugation in roseobacters is QS-controlled and that the QS system may detect a wide array of long-chain AHLs at the cell surface.

  1. Peptides Derived from Type IV Collagen, CXC Chemokines, and Thrombospondin-1 Domain-Containing Proteins Inhibit Neovascularization and Suppress Tumor Growth in MDA-MB-231 Breast Cancer Xenografts

    Directory of Open Access Journals (Sweden)

    Jacob E. Koskimaki

    2009-12-01

    Full Text Available Angiogenesis or neovascularization, the process of new blood vessel formation from preexisting microvasculature, involves interactions among several cell types including parenchymal, endothelial cells, and immune cells. The formation of new vessels is tightly regulated by a balance between endogenous proangiogenic and antiangiogenic factors to maintain homeostasis in tissue; tumor progression and metastasis in breast cancer have been shown to be angiogenesis-dependent. We previously introduced a systematic methodology to identify putative endogenous antiangiogenic peptides and validated these predictions in vitro in human umbilical vein endothelial cell proliferation and migration assays. These peptides are derived from several protein families including type IV collagen, CXC chemokines, and thrombospondin-1 domain-containing proteins. On the basis of the results from the in vitro screening, we have evaluated the ability of one peptide selected from each family named pentastatin-1, chemokinostatin-1, and properdistatin, respectively, to suppress angiogenesis in an MDA-MB-231 human breast cancer orthotopic xenograft model in severe combined immunodeficient mice. Peptides were administered intraperitoneally once per day. We have demonstrated significant suppression of tumor growth in vivo and subsequent reductions in microvascular density, indicating the potential of these peptides as therapeutic agents for breast cancer.

  2. Fratura extra-articular da extremidade medial da clavícula associada à luxação acromioclavicular tipo IV: relato de caso Extra-articular fracture of the medial end of the clavicle associated with type IV acromioclavicular dislocation: case report

    Directory of Open Access Journals (Sweden)

    Mário Chaves Correa

    2011-10-01

    Full Text Available Ocorrendo isoladamente, as fraturas da clavícula e as luxações acromioclaviculares são lesões muito comuns. A combinação de uma luxação acromioclavicular e de uma fratura do terço lateral da clavícula não é rara. Entretanto, existem muito poucos casos descritos de luxações acromioclaviculares associadas a fraturas do terço médio da clavícula; aquelas associadas a fraturas do terço medial são ainda mais raras. Nós reportamos o caso de um indivíduo adulto do sexo masculino que sofreu uma luxação acromioclavicular (tipo IV associada a uma fratura extra-articular desviada da extremidade medial da clavícula (grupo 3 de Almann em um acidente ciclístico. O paciente foi tratado na fase aguda com redução aberta e fixação interna das duas lesões. Na avaliação clínica, 12 meses após a cirurgia, o paciente apresentava-se assintomático, com mobilidade ativa e passiva completa, força e resistência normais e simetria das cinturas escapulares. As radiografias e a tomografia computadorizada tridimensional mostravam subluxação posterossuperior persistente da articulação acromioclavicular e consolidação anatômica da fratura clavicular.Fractures of the clavicle and acromioclavicular dislocations are very common injuries when they occur separately. The combination of an acromioclavicular dislocation and a fracture of the lateral third of the clavicle is not rare. However, there are very few reported cases of acromioclavicular dislocations associated with fractures of the middle third of the clavicle; those associated with fractures of the medial third are even rarer. We report the case of an adult male who suffered an acromioclavicular dislocation (type IV associated with a displaced extra-articular fracture of the medial end of the clavicle (Almann group 3 in a cycling accident. The patient was treated during the acute phase with open reduction and internal fixation of the two lesions. At the clinical evaluation 12

  3. IV access in dental practice.

    LENUS (Irish Health Repository)

    Fitzpatrick, J J

    2009-04-01

    Intravenous (IV) access is a valuable skill for dental practitioners in emergency situations and in IV sedation. However, many people feel some apprehension about performing this procedure. This article explains the basic principles behind IV access, and the relevant anatomy and physiology, as well as giving a step-by-step guide to placing an IV cannula.

  4. Dibromidodimethyldipyridineplatinum(IV

    Directory of Open Access Journals (Sweden)

    Mairéad E. Kelly

    2008-11-01

    Full Text Available In the title complex, [PtBr2(CH32(C5H5N2], the PtIV metal centre lies on a twofold rotation axis and adopts a slightly distorted octahedral coordination geometry. The structure displays weak intramolecular C—H...Br hydrogen-bonding interactions.

  5. Infantile mitochondrial hepatopathy is a cardinal feature of MEGDEL syndrome (3-methylglutaconic aciduria type IV with sensorineural deafness, encephalopathy and Leigh-like syndrome) caused by novel mutations in SERAC1.

    Science.gov (United States)

    Sarig, Ofer; Goldsher, Dorit; Nousbeck, Janna; Fuchs-Telem, Dana; Cohen-Katsenelson, Ksenya; Iancu, Theodore C; Manov, Irena; Saada, Ann; Sprecher, Eli; Mandel, Hanna

    2013-09-01

    3-Methylglutaconic aciduria (3-MGCA) type IV is defined as a heterogeneous group of inborn errors featuring in common 3-MGCA and associated with primary mitochondrial dysfunction leading to a spectrum of multisystem conditions. We studied four patients who presented at birth with a clinical picture simulating a primary mitochondrial hepatic disorder consistent with the MEGDEL syndrome including 3-MGCA, sensorineural deafness, encephalopathy and a brain magnetic resonance imaging with signs of Leigh disease. All affected children displayed biochemical features consistent with mitochondrial OXPHOS dysfunction including hepatic mitochondrial DNA depletion in one patient. Homozygosity mapping identified a candidate locus on 6q25.2-6q26. Using whole exome sequencing, we identified two novel homozygous mutations in SERAC1 recently reported to harbor mutations in MEGDEL syndrome. Both mutations were found to lead to decreased or absent expression of SERAC1. The present findings indicate that infantile hepatopathy is a cardinal feature of MEGDEL syndrome. We thus propose to rename the disease MEGDHEL syndrome. Copyright © 2013 Wiley Periodicals, Inc.

  6. (2R)-4-Oxo-4[3-(Trifluoromethyl)-5,6-diihydro:1,2,4}triazolo[4,3-a}pyrazin-7(8H)-y1]-1-(2,4,5-trifluorophenyl)butan-2-amine: A Potent, Orally Active Dipeptidyl Peptidase IV Inhibitor for the Treatment of Type 2 Diabetes

    Energy Technology Data Exchange (ETDEWEB)

    Kim, D.; Wang, L.; Beconi, M.; Eiermann, G.; Fisher, M.; He, H.; Hickey, G.; Kowalchick, Jennifer; Leiting, Barbara; Lyons, K.; Marsilio, F.; McCann, F.; Patel, R.; Petrov, A.; Scapin, G.; Patel, S.; Roy, R.; Wu, J.; Wyvratt, M.; Zhang, B.; Zhu, L.; Thornberry, N.; Weber, A. (Merck)

    2010-11-10

    A novel series of {beta}-amino amides incorporating fused heterocycles, i.e., triazolopiperazines, were synthesized and evaluated as inhibitors of dipeptidyl peptidase IV (DPP-IV) for the treatment of type 2 diabetes. (2R)-4-Oxo-4-[3-(trifluoromethyl)-5,6-dihydro[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl]-1-(2,4,5-trifluorophenyl)butan-2-amine (1) is a potent, orally active DPP-IV inhibitor (IC{sub 50} = 18 nM) with excellent selectivity over other proline-selective peptidases, oral bioavailability in preclinical species, and in vivo efficacy in animal models. MK-0431, the phosphate salt of compound 1, was selected for development as a potential new treatment for type 2 diabetes.

  7. A sputnik IV saga

    Science.gov (United States)

    Lundquist, Charles A.

    2009-12-01

    The Sputnik IV launch occurred on May 15, 1960. On May 19, an attempt to deorbit a 'space cabin' failed and the cabin went into a higher orbit. The orbit of the cabin was monitored and Moonwatch volunteer satellite tracking teams were alerted to watch for the vehicle demise. On September 5, 1962, several team members from Milwaukee, Wisconsin made observations starting at 4:49 a.m. of a fireball following the predicted orbit of Sputnik IV. Requests went out to report any objects found under the fireball path. An early morning police patrol in Manitowoc had noticed a metal object on a street and had moved it to the curb. Later the officers recovered the object and had it dropped off at the Milwaukee Journal. The Moonwarch team got the object and reported the situation to Moonwatch Headquarters at the Smithsonian Astrophysical Observatory. A team member flew to Cambridge with the object. It was a solid, 9.49 kg piece of steel with a slag-like layer attached to it. Subsequent analyses showed that it contained radioactive nuclei produced by cosmic ray exposure in space. The scientists at the Observatory quickly recognized that measurements of its induced radioactivity could serve as a calibration for similar measurements of recently fallen nickel-iron meteorites. Concurrently, the Observatory directorate informed government agencies that a fragment from Sputnik IV had been recovered. Coincidently, a debate in the UN Committee on Peaceful Uses of Outer Space involved the issue of liability for damage caused by falling satellite fragments. On September 12, the Observatory delivered the bulk of the fragment to the US Delegation to the UN. Two days later, the fragment was used by US Ambassador Francis Plimpton as an exhibit that the time had come to agree on liability for damage from satellite debris. He offered the Sputnik IV fragment to USSR Ambassador P.D. Morozov, who refused the offer. On October 23, Drs. Alla Massevitch and E.K. Federov of the USSR visited the

  8. Molecular characterization of spa type t127, sequence type 1 methicillin-resistant Staphylococcus aureus from pigs.

    Science.gov (United States)

    Franco, Alessia; Hasman, Henrik; Iurescia, Manuela; Lorenzetti, Raniero; Stegger, Marc; Pantosti, Annalisa; Feltrin, Fabiola; Ianzano, Angela; Porrero, Maria Concepción; Liapi, Maria; Battisti, Antonio

    2011-06-01

    The aim of this study was to provide molecular characterization of methicillin-resistant Staphylococcus aureus (MRSA) spa type t127, sequence type (ST) 1 isolates, detected in a European baseline survey in holdings of breeding pigs, to determine phenotypic and genotypic drug resistance and to compare the results with those obtained from a collection of t127, ST1 MRSA and methicillin-susceptible S. aureus (MSSA) clinical isolates. Twenty-four t127, ST1 MRSA from dust sampled in different breeding holdings in Italy, Spain and Cyprus were studied, along with 2 t127, ST1 MRSA from fattening pigs and 11 human t127, ST1 MRSA and MSSA. Genotyping was performed using multilocus sequence typing (MLST), spa typing and PFGE. SCCmec elements were characterized by multiplex-PCR and resistance and pathogenicity genes by PCR and microarray. PFGE patterns separated a porcine cluster (PC) from a human cluster (HC), with 75% similarity. The PC carried SCCmec cassette type V, while all isolates of the HC carried SCCmec cassette type IVa. Kanamycin resistance mediated by aadD, fluoroquinolone and erm(A)-mediated macrolide resistance and the absence of the sakA gene were features of the PC only. All isolates of both clusters were positive for LukE-LukD and LuF-LukS-HlgA leukotoxin genes and one human MSSA harboured Panton-Valentine leucocidin genes. Despite differences in the host-specific genetic features, the possibility of PC transmission to humans cannot be excluded. MRSA spa type t127, ST1 from pigs possesses several virulence and resistance genes towards major classes of antimicrobials and may represent a serious therapeutic challenge in case of invasive infections in humans.

  9. Predicting DPP-IV inhibitors with machine learning approaches

    Science.gov (United States)

    Cai, Jie; Li, Chanjuan; Liu, Zhihong; Du, Jiewen; Ye, Jiming; Gu, Qiong; Xu, Jun

    2017-04-01

    Dipeptidyl peptidase IV (DPP-IV) is a promising Type 2 diabetes mellitus (T2DM) drug target. DPP-IV inhibitors prolong the action of glucagon-like peptide-1 (GLP-1) and gastric inhibitory peptide (GIP), improve glucose homeostasis without weight gain, edema, and hypoglycemia. However, the marketed DPP-IV inhibitors have adverse effects such as nasopharyngitis, headache, nausea, hypersensitivity, skin reactions and pancreatitis. Therefore, it is still expected for novel DPP-IV inhibitors with minimal adverse effects. The scaffolds of existing DPP-IV inhibitors are structurally diversified. This makes it difficult to build virtual screening models based upon the known DPP-IV inhibitor libraries using conventional QSAR approaches. In this paper, we report a new strategy to predict DPP-IV inhibitors with machine learning approaches involving naïve Bayesian (NB) and recursive partitioning (RP) methods. We built 247 machine learning models based on 1307 known DPP-IV inhibitors with optimized molecular properties and topological fingerprints as descriptors. The overall predictive accuracies of the optimized models were greater than 80%. An external test set, composed of 65 recently reported compounds, was employed to validate the optimized models. The results demonstrated that both NB and RP models have a good predictive ability based on different combinations of descriptors. Twenty "good" and twenty "bad" structural fragments for DPP-IV inhibitors can also be derived from these models for inspiring the new DPP-IV inhibitor scaffold design.

  10. The abcEDCBA-Encoded ABC Transporter and the virB Operon-Encoded Type IV Secretion System of Brucella ovis Are Critical for Intracellular Trafficking and Survival in Ovine Monocyte-Derived Macrophages.

    Directory of Open Access Journals (Sweden)

    Auricelio A Macedo

    Full Text Available Brucella ovis infection is associated with epididymitis, orchitis and infertility in rams. Most of the information available on B. ovis and host cell interaction has been generated using murine macrophages or epithelial cell lines, but the interaction between B. ovis and primary ovine macrophages has not been studied. The aim of this study was to evaluate the role of the B. ovis abcEDCBA-encoded ABC transporter and the virB operon-encoded Type IV Secretion System (T4SS during intracellular survival of B. ovis in ovine peripheral blood monocyte-derived macrophages. ΔabcBA and ΔvirB2 mutant strains were unable to survive in the intracellular environment when compared to the WT B. ovis at 48 hours post infection (hpi. In addition, these mutant strains cannot exclude the lysosomal marker LAMP1 from its vacuolar membrane, and their vacuoles do not acquire the endoplasmic reticulum marker calreticulin, which takes place in the WT B. ovis containing vacuole. Higher levels of nitric oxide production were observed in macrophages infected with WT B. ovis at 48 hpi when compared to macrophages infected with the ΔabcBA or ΔvirB2 mutant strains. Conversely, higher levels of reactive oxygen species were detected in macrophages infected with the ΔabcBA or ΔvirB2 mutant strains at 48 hpi when compared to macrophages infected with the WT strain. Our results demonstrate that B. ovis is able to persist and multiply in ovine macrophages, while ΔabcBA and ΔvirB2 mutations prevent intracellular multiplication, favor phagolysosome fusion, and impair maturation of the B. ovis vacuole towards an endoplasmic reticulum-derived compartment.

  11. Equilibrium between Different Coordination Geometries in Oxidovanadium(IV) Complexes

    Science.gov (United States)

    Ugone, Valeria; Garribba, Eugenio; Micera, Giovanni; Sanna, Daniele

    2015-01-01

    In this laboratory activity, the equilibrium between square pyramidal and octahedral V(IV)O[superscript 2+] complexes is described. We propose a set of experiments to synthesize and characterize two types of V(IV)O[superscript 2+] complexes. The experiment allows great flexibility and may be effectively used at a variety of levels and the activity…

  12. Immunohistochemical investigation of alpha1 (IV) and alpha5 (IV) collagen chains in a broad spectrum of melanocytic tumours.

    Science.gov (United States)

    Quatresooz, Pascale; Piérard, Gérald E

    2005-06-01

    Cells of melanocytic naevi and cutaneous malignant melanomas (MM) are surrounded by a basement membrane (BM). To scrutinize any difference between the deposits of alpha1 (IV) and alpha5 (IV) collagen chains in melanocytic naevi and MM. A total of 27 common melanocytic naevi, 11 dysplastic naevi, 21 atypical naevi (melanocytomas) including Spitz and non-Spitz types, as well as 24 MM were studied. Their phenotypic and functional characteristics defined by immunohistochemistry using a panel of antibodies, including those directed to the alpha1 (IV), alpha3 (IV) and alpha5 (IV) collagen chains. Almost all naevi and half the melanocytomas exhibited a strong positivity for the alpha1 (IV) collagen chain. By contrast, the remaining melanocytomas and MM presented a heterogeneous staining pattern for the alpha1 (IV) collagen chain. One third of the naevi, 23% of the MM without cutaneous micrometastasis and 83% of MM with cutaneous micrometastasis showed discrete cytoplasmic positivity for the alpha5 (IV) collagen chain. All other melanocytic tumours were negative for this antibody. Rare MM cells in transepidermal migration were stained with the anti-alpha1 (IV) or alpha5 (IV) collagen chain antibodies. No immunoreactivity for the alpha3 (IV) collagen chain was disclosed in any of the samples. We report the expression of alpha1 (IV) and alpha5 (IV) collagen chains in naevi and MM. The inconsistent staining pattern for alpha1 (IV) collagen chain in phenotypically atypical melanocytomas and in MM highlight the heterogeneity in both cell differentiation and stroma-tumour interactions. This biological aspect may be related to neoplastic progression and influence metastatic potential.

  13. Type IV Procollagen Missense Mutations Associated With Defects of the Eye, Vascular Stability, the Brain, Kidney Function and Embryonic or Postnatal Viability in the Mouse, Mus musculus: An Extension of the Col4a1 Allelic Series and the Identification of the First Two Col4a2 Mutant Alleles

    Science.gov (United States)

    Favor, Jack; Gloeckner, Christian Johannes; Janik, Dirk; Klempt, Martina; Neuhäuser-Klaus, Angelika; Pretsch, Walter; Schmahl, Wolfgang; Quintanilla-Fend, Leticia

    2007-01-01

    The basement membrane is important for proper tissue development, stability, and physiology. Major components of the basement membrane include laminins and type IV collagens. The type IV procollagens Col4a1 and Col4a2 form the heterotrimer [α1(IV)]2[α2(IV)], which is ubiquitously expressed in basement membranes during early developmental stages. We present the genetic, molecular, and phenotypic characterization of nine Col4a1 and three Col4a2 missense mutations recovered in random mutagenesis experiments in the mouse. Heterozygous carriers express defects in the eye, the brain, kidney function, vascular stability, and viability. Homozygotes do not survive beyond the second trimester. Ten mutations result in amino acid substitutions at nine conserved Gly sites within the collagenous domain, one mutation is in the carboxy-terminal noncollagenous domain, and one mutation is in the signal peptide sequence and is predicted to disrupt the signal peptide cleavage site. Patients with COL4A2 mutations have still not been identified. We suggest that the spontaneous intraorbital hemorrhages observed in the mouse are a clinically relevant phenotype with a relatively high predictive value to identify carriers of COL4A1 or COL4A2 mutations. PMID:17179069

  14. Diaquatetrabromidotin(IV trihydrate

    Directory of Open Access Journals (Sweden)

    Fei Ye

    2012-09-01

    Full Text Available The title compound, [SnBr4(H2O2]·3H2O, forms large colourless crystals in originally sealed samples of tin tetrabromide. It constitutes the first structurally characterized hydrate of SnBr4 and is isostructural with the corresponding hydrate of SnCl4. It is composed of SnIV atoms octahedrally coordinated by four Br atoms and two cis-related water molecules. The octahedra exhibit site symmetry 2. They are arranged into columns along [001] via medium–strong O—H...O hydrogen bonds involving the two lattice water molecules (one situated on a twofold rotation axis while the chains are interconnected via longer O—H...Br hydrogen bonds, forming a three-dimensional network.

  15. Estabilidade dimensional dos moldes de poliéter e polissulfeto obtidos com a técnica do casquete de acrílico, vazados em gesso pedra especial tipo IV e V = Dimensional stability of polyether and polysulfide impression materials obtained with acrylic coping techniques pouring in stone type IV and V

    Directory of Open Access Journals (Sweden)

    Alves, Maria Cechinel

    2005-01-01

    Full Text Available O propósito deste estudo foi verificar o comportamento de moldes de polissulfeto (Permlastic e poliéter (Impregum Soft obtidos por meio da técnica do casquete de resina acrílica. Avaliou-se a estabilidade dimensional dos moldes confeccionados de uma matriz metálica contendo dois preparos protéticos, com diâmetros e alturas diferentes. Os moldes de cada material foram vazados com gessos pedra IV (Vel-mix e gesso pedra V (Exadur, perfazendo um total de quatro combinações experimentais: (polissulfeto/gesso-pedra IV, polissulfeto/gesso-pedra V, poliéter/gesso-pedra IV e poliéter/gesso-pedra V. Os troquéis de gesso foram mensurados com um paquímetro digital, 24 horas após sua obtenção. Os resultados obtidos foram submetidos a ANOVA a 5%. Pode-se concluir que os moldes de poliéter vazados com gesso pedra tipo V apresentaram melhor estabilidade dimensional em relação às outras condições experimentais

  16. Errors Associated with IV Infusions in Critical Care

    National Research Council Canada - National Science Library

    Summa-Sorgini, Claudia; Fernandes, Virginia; Lubchansky, Stephanie; Mehta, Sangeeta; Hallett, David; Bailie, Toni; Lapinsky, Stephen E; Burry, Lisa

    2012-01-01

    ...% of medication errors. To determine the type and frequency of errors associated with prescribing, documenting, and administering IV infusions, and to also determine if a correlation exists between the incidence of errors...

  17. Dioxobridged complexes of molybdenum (IV) and tungsten (IV) with ...

    Indian Academy of Sciences (India)

    Abstract. Six new dioxobridged complexes of molybdenum (IV) and tungsten (IV) with N-alkylphenothiazines having the general formula M2O4(L)2(H2O)2 [where M = molybdenum or tungsten and L = N-alkylphenothiazines] have been synthesised. The complexes have been characterised on the basis of analytical, molar ...

  18. Estabilidade dimensional dos moldes de poliéter e polissulfeto obtidos com a técnica do casquete de acrílico, vazados em gesso pedra especial tipo IV e V = Dimensional stability of polyether and polysulfide impression materials obtained with acrylic coping techniques pouring in stone type IV and V

    OpenAIRE

    Alves, Maria Cechinel; Soares, Carlos Renato; Zani, Izo Milton

    2005-01-01

    O propósito deste estudo foi verificar o comportamento de moldes de polissulfeto (Permlastic) e poliéter (Impregum Soft) obtidos por meio da técnica do casquete de resina acrílica. Avaliou-se a estabilidade dimensional dos moldes confeccionados de uma matriz metálica contendo dois preparos protéticos, com diâmetros e alturas diferentes. Os moldes de cada material foram vazados com gessos pedra IV (Vel-mix) e gesso pedra V (Exadur), perfazendo um total de quatro combinações experimentais: (pol...

  19. Topological characterisation and identification of critical domains within glucosyltransferase IV (GtrIV of Shigella flexneri

    Directory of Open Access Journals (Sweden)

    Nair Anesh

    2011-12-01

    Full Text Available Abstract Background The three bacteriophage genes gtrA, gtrB and gtr(type are responsible for O-antigen glucosylation in Shigella flexneri. Both gtrA and gtrB have been demonstrated to be highly conserved and interchangeable among serotypes while gtr(type was found to be specific to each serotype, leading to the hypothesis that the Gtr(type proteins are responsible for attaching glucosyl groups to the O-antigen in a site- and serotype- specific manner. Based on the confirmed topologies of GtrI, GtrII and GtrV, such interaction and attachment of the glucosyl groups to the O-antigen has been postulated to occur in the periplasm. Results In this study, the topology of GtrIV was experimentally determined by creating different fusions between GtrIV and a dual-reporter protein, PhoA/LacZ. This study shows that GtrIV consists of 8 transmembrane helices, 2 large periplasmic loops, 2 small cytoplasmic N- and C- terminal ends and a re-entrant loop that occurs between transmembrane helices III and IV. Though this topology differs from that of GtrI, GtrII, GtrV and GtrX, it is very similar to that of GtrIc. Furthermore, both the N-terminal periplasmic and the C-terminal periplasmic loops are important for GtrIV function as shown via a series of loop deletion experiments and the creation of chimeric proteins between GtrIV and its closest structural homologue, GtrIc. Conclusion The current study provides the basis for elucidating the structure and mechanism of action of this important O-antigen modifying glucosyltransferase.

  20. Prevalence of methicillin-resistant Staphylococcus aureus (MRSA) infection and the molecular characteristics of MRSA bacteraemia over a two-year period in a tertiary teaching hospital in Malaysia.

    Science.gov (United States)

    Sit, Pik San; Teh, Cindy Shuan Ju; Idris, Nuryana; Sam, I-Ching; Syed Omar, Sharifah Faridah; Sulaiman, Helmi; Thong, Kwai Lin; Kamarulzaman, Adeeba; Ponnampalavanar, Sasheela

    2017-04-13

    Methicillin-resistant Staphylococcus aureus (MRSA) is an established pathogen that causes hospital- and community-acquired infections worldwide. The prevalence rate of MRSA infections were reported to be the highest in Asia. As there is limited epidemiological study being done in Malaysia, this study aimed to determine the prevalence of MRSA infection and the molecular characteristics of MRSA bacteraemia. Two hundred and nine MRSA strains from year 2011 to 2012 were collected from a tertiary teaching hospital in Malaysia. The strains were characterized by antimicrobial susceptibility testing, staphylococcal cassette chromosome mec (SCCmec) typing, detection of Panton-Valentine leukocidin (PVL) gene, multilocus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE). Patient's demographic and clinical data were collected and correlated with molecular data by statistical analysis. Male gender and patient >50 years of age (p MRSA acquisition. Fifty-nine percent of MRSA strains were HA-MRSA that carried SCCmec type II, III, IV and V while 31% were CA-MRSA strains with SCCmec III, IV and V. The prevalence of PVL gene among 2011 MRSA strains was 5.3% and no PVL gene was detected in 2012 MRSA strains. All of the strains were sensitive to vancomycin. However, vancomycin MIC creep phenomenon was demonstrated by the increased number of MRSA strains with MIC ≥1.5 μg/mL (p = 0.008) between 2011 and 2012. Skin disease (p = 0.034) and SCCmec type III (p = 0.0001) were found to be significantly associated with high vancomycin MIC. Forty-four percent of MRSA strains from blood, were further subtyped by MLST and PFGE. Most of the bacteraemia cases were primary bacteraemia and the common comorbidities were diabetes, hypertension and chronic kidney disease. The predominant pulsotype was pulsotype C exhibited by SCCmec III-ST239. This is a first study in Malaysia that reported the occurrence of MRSA clones such as SCCmec V-ST5, untypeable-ST508, SCCmec IV-ST1

  1. Zirconium(IV) and Hafnium(IV)-Catalyzed Highly Enantioselective Epoxidation of Homoallylic and Bishomoallylic Alcohols

    Science.gov (United States)

    Li, Zhi; Yamamoto, Hisashi

    2010-01-01

    In this report, zirconium(IV) and hafnium(IV)-bishydroxamic acid complexes were utilized in the highly enantioselective epoxidation of homoallylic alcohols and bishomoallylic alcohols, which used to be quite difficult substrates for other types of asymmetric epoxidation reactions. The performance of the catalyst was improved by adding polar additive and molecular sieves. For homoallylic alcohols, the reaction could provide epoxy alcohols in up to 81% yield and up to 98% ee, while for bishomoallylic alcohols, up to 75% yield and 99% ee of epoxy alcohols rather than cyclize compounds could be obtained in most cases. PMID:20481541

  2. Zirconium(IV)- and hafnium(IV)-catalyzed highly enantioselective epoxidation of homoallylic and bishomoallylic alcohols.

    Science.gov (United States)

    Li, Zhi; Yamamoto, Hisashi

    2010-06-16

    In this report, zirconium(IV)- and hafnium(IV)-bishydroxamic acid complexes were utilized in the highly enantioselective epoxidation of homoallylic alcohols and bishomoallylic alcohols, which used to be quite difficult substrates for other types of asymmetric epoxidation reactions. The performance of the catalyst was improved by adding polar additive and molecular sieves. For homoallylic alcohols, the reaction could provide epoxy alcohols in up to 83% yield and up to 98% ee, while, for bishomoallylic alcohols, up to 79% yield and 99% ee of epoxy alcohols rather than cyclized tetrahydrofuran compounds could be obtained in most cases.

  3. Synthesis and characterization of thorium(IV) and uranium(IV) complexes with Schiff bases

    Energy Technology Data Exchange (ETDEWEB)

    Radoske, Thomas; Maerz, Juliane; Kaden, Peter; Patzschke, Michael; Ikeda-Ohno, Atsushi [Helmholtz-Zentrum Dresden-Rossendorf e.V., Dresden (Germany). Chemistry of the F-Elements

    2017-06-01

    We report herein the synthesis and characterization of several imine complexes of tetravalent thorium (Th(IV)) and uranium (U(IV)). The ligands investigated in this study are a Schiff base type, including the well-known salen ligand (H{sub 2}Le, Fig. 1). The complexation in solution was investigated by NMR measurements indicating paramagnetic effects of unpaired f-electrons of U(IV) on the ligand molecule. We also determ