OmpA influences Escherichia coli biofilm formation by repressing cellulose production through the CpxRA two-component system.

PubMed

Ma, Qun; Wood, Thomas K

2009-10-01

Previously we discovered that OmpA of Escherichia coli increases biofilm formation on polystyrene surfaces (González Barrios et al., Biotechnol Bioeng, 93:188-200, 2006a). Here we show OmpA influences biofilm formation differently on hydrophobic and hydrophilic surfaces since it represses cellulose production which is hydrophilic. OmpA increased biofilm formation on polystyrene, polypropylene, and polyvinyl surfaces while it decreased biofilm formation on glass surfaces. Sand column assays corroborated that OmpA decreases attachment to hydrophilic surfaces. The ompA mutant formed sticky colonies, and the extracellular polysaccharide that caused stickiness was identified as cellulose. A whole-transcriptome study revealed that OmpA induces the CpxRA two-component signal transduction pathway that responds to membrane stress. CpxA phosphorylates CpxR and results in reduced csgD expression. Reduced CsgD production represses adrA expression and results in reduced cellulose production since CsgD and AdrA are responsible for 3,5-cyclic diguanylic acid and cellulose synthesis. Real-time polymerase chain reaction confirmed csgD and adrA are repressed by OmpA. Biofilm and cellulose assays with double deletion mutants adrA ompA, csgB ompA, and cpxR ompA confirmed OmpA decreased cellulose production and increased biofilm formation on polystyrene surfaces through CpxR and AdrA. Further evidence of the link between OmpA and the CpxRA system was that overproduction of OmpA disrupted the membrane and led to cell lysis. Therefore, OmpA inhibits cellulose production through the CpxRA stress response system, and this reduction in cellulose increases biofilm formation on hydrophobic surfaces.

A predictive multi-linear regression model for organic micropollutants, based on a laboratory-scale column study simulating the river bank filtration process.

PubMed

Bertelkamp, C; Verliefde, A R D; Reynisson, J; Singhal, N; Cabo, A J; de Jonge, M; van der Hoek, J P

2016-03-05

This study investigated relationships between OMP biodegradation rates and the functional groups present in the chemical structure of a mixture of 31 OMPs. OMP biodegradation rates were determined from lab-scale columns filled with soil from RBF site Engelse Werk of the drinking water company Vitens in The Netherlands. A statistically significant relationship was found between OMP biodegradation rates and the functional groups of the molecular structures of OMPs in the mixture. The OMP biodegradation rate increased in the presence of carboxylic acids, hydroxyl groups, and carbonyl groups, but decreased in the presence of ethers, halogens, aliphatic ethers, methyl groups and ring structures in the chemical structure of the OMPs. The predictive model obtained from the lab-scale soil column experiment gave an accurate qualitative prediction of biodegradability for approximately 70% of the OMPs monitored in the field (80% excluding the glymes). The model was found to be less reliable for the more persistent OMPs (OMPs with predicted biodegradation rates lower or around the standard error=0.77d(-1)) and OMPs containing amide or amine groups. These OMPs should be carefully monitored in the field to determine their removal during RBF. Copyright © 2015 Elsevier B.V. All rights reserved.

Comparative proteomic analysis of outer membrane protein 43 (omp43)-deficient Bartonella henselae.

PubMed

Kang, Jun-Gu; Lee, Hee-Woo; Ko, Sungjin; Chae, Joon-Seok

2018-01-31

Outer membrane proteins (OMPs) of Gram-negative bacteria constitute the first line of defense protecting cells against environmental stresses including chemical, biophysical, and biological attacks. Although the 43-kDa OMP (OMP43) is major porin protein among Bartonella henselae -derived OMPs, its function remains unreported. In this study, OMP43-deficient mutant B. henselae (Δomp43) was generated to investigate OMP43 function. Interestingly, Δ omp 43 exhibited weaker proliferative ability than that of wild-type (WT) B. henselae . To study the differences in proteomic expression between WT and Δ omp 43, two-dimensional gel electrophoresis-based proteomic analysis was performed. Based on Clusters of Orthologus Groups functional assignments, 12 proteins were associated with metabolism, 7 proteins associated with information storage and processing, and 3 proteins associated with cellular processing and signaling. By semi-quantitative reverse transcriptase polymerase chain reaction, increases in tld D, efp, ntr X, pdh A, pur B, and ATPA mRNA expression and decreases in Rho and yfe A mRNA expression were confirmed in Δ omp 43. In conclusion, this is the first report showing that a loss of OMP43 expression in B. henselae leads to retarded proliferation. Furthermore, our proteomic data provide useful information for the further investigation of mechanisms related to the growth of B. henselae.

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

PubMed

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

2017-06-01

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

Antibody Reactivity to Omp31 from Brucella melitensis in Human and Animal Infections by Smooth and Rough Brucellae

PubMed Central

Cassataro, Juliana; Pasquevich, Karina; Bruno, Laura; Wallach, Jorge C.; Fossati, Carlos A.; Baldi, Pablo C.

2004-01-01

Group 3 of outer membrane proteins (OMPs) of Brucella includes Omp25 and Omp31, which share 34% identity. Omp25 is highly conserved in Brucella species, and Omp31 is present in all Brucella species, except Brucella abortus. Antibodies to Brucella melitensis Omp31 have been sought only in infected sheep, and Western blotting of sera from infected sheep did not reveal anti-Omp31 reactivity. We obtained recombinant purified Omp31 (B. melitensis) and tested its recognition by sera from humans and animals suffering from brucellosis by an indirect enzyme-linked immunosorbent assay (ELISA). Serum samples from 74 patients, 57 sheep, and 47 dogs were analyzed; brucellosis was confirmed by bacteriological isolation in all ovine and canine cases and 31 human cases of brucellosis. Thirty-five patients (47%) were positive for antibodies to Omp31, including seven cases of Brucella suis infection, two cases of B. abortus infection, and three cases of B. melitensis infection. Of 39 sheep naturally infected with B. melitensis (biovars 1 and 3), 23 (59%) were positive for antibodies to Omp31. Anti-Omp31 antibodies were also detected in 12 of 18 rams (67%) in which Brucella ovis was isolated from semen. Antibodies to Omp31 were also found in 41 (87%) of the 47 dogs, including 13 with recent infection. These results suggest that an indirect ELISA using recombinant purified Omp31 from B. melitensis would be of limited value for the diagnosis of human and animal brucellosis. Nevertheless, the potential usefulness of this antigen in combination with other recombinant proteins from Brucella should not be dismissed.   PMID:14715555

Structural and dynamical properties of the porins OmpF and OmpC: insights from molecular simulations

NASA Astrophysics Data System (ADS)

Kumar, Amit; Hajjar, Eric; Ruggerone, Paolo; Ceccarelli, Matteo

2010-11-01

In this paper we investigate the structural and dynamical properties of the two major porins (OmpF and OmpC) in Escherichia coli, using molecular dynamics (MD) simulations. In particular we characterized the atomic fluctuations, correlated motions, temperature dependence, solvent-accessible cross-sectional area and water dynamics in the key regions of the two channels. Our in-depth analysis allows us to highlight the importance of both the key conserved and substituted residues between OmpF and OmpC. The latter is characterized by a narrower and longer constriction region with respect to OmpF. OmpC also showed a higher stability upon increasing temperature. We then present the results of transport properties by using accelerated MD simulations to probe the diffusion of norfloxacin (a fluoroquinolone antibiotic) through the two porins OmpF/OmpC. Our study constitutes a step forward towards understanding the structure-function relationship of the two porins' channels. This will benefit the research of antibacterials with improved permeation properties and nanopores that aim to use these porins as sensing systems.

Estimating the budget impact of orphan drugs in Sweden and France 2013–2020

PubMed Central

2014-01-01

Background The growth in expenditure on orphan medicinal products (OMP) across Europe has been identified as a concern. Estimates of future expenditure in Europe have suggested that OMPs could account for a significant proportion of total pharmaceutical expenditure in some countries, but few of these forecasts have been well validated. This analysis aims to establish a robust forecast of the future budget impact of OMPs on the healthcare systems in Sweden and France. Methods A dynamic forecasting model was created to estimate the budget impact of OMPs in Sweden and France between 2013 and 2020. The model used historical data on OMP designation and approval rates to predict the number of new OMPs coming to the market. Average OMP sales were estimated for each year post-launch by regression analysis of historical sales data. Total forecast sales were compared with expected sales of all pharmaceuticals in each country to quantify the relative budget impact. Results The model predicts that by 2020, 152 OMPs will have marketing authorization in Europe. The base case OMP budget impacts are forecast to grow from 2.7% in Sweden and 3.2% in France of total drug expenditure in 2013 to 4.1% in Sweden and 4.9% in France by 2020. The principal driver of expenditure growth is the number of new OMPs obtaining OMP designation. This is tempered by the slowing success rate for new approvals and the loss of intellectual property protection on existing orphan medicines. Given the forward-looking nature of the analysis, uncertainty exists around model parameters and sensitivity analysis found peak year budget impact varying between 2% and 11%. Conclusion The budget impact of OMPs in Sweden and France is likely to remain sustainable over time and a relatively small proportion of total pharmaceutical expenditure. This forecast could be affected by changes in the success rate for OMP approvals, average cost of OMPs, and the type of companies developing OMPs. PMID:24524281

Estimating the budget impact of orphan drugs in Sweden and France 2013-2020.

PubMed

Hutchings, Adam; Schey, Carina; Dutton, Richard; Achana, Felix; Antonov, Karolina

2014-02-13

The growth in expenditure on orphan medicinal products (OMP) across Europe has been identified as a concern. Estimates of future expenditure in Europe have suggested that OMPs could account for a significant proportion of total pharmaceutical expenditure in some countries, but few of these forecasts have been well validated. This analysis aims to establish a robust forecast of the future budget impact of OMPs on the healthcare systems in Sweden and France. A dynamic forecasting model was created to estimate the budget impact of OMPs in Sweden and France between 2013 and 2020. The model used historical data on OMP designation and approval rates to predict the number of new OMPs coming to the market. Average OMP sales were estimated for each year post-launch by regression analysis of historical sales data. Total forecast sales were compared with expected sales of all pharmaceuticals in each country to quantify the relative budget impact. The model predicts that by 2020, 152 OMPs will have marketing authorization in Europe. The base case OMP budget impacts are forecast to grow from 2.7% in Sweden and 3.2% in France of total drug expenditure in 2013 to 4.1% in Sweden and 4.9% in France by 2020. The principal driver of expenditure growth is the number of new OMPs obtaining OMP designation. This is tempered by the slowing success rate for new approvals and the loss of intellectual property protection on existing orphan medicines. Given the forward-looking nature of the analysis, uncertainty exists around model parameters and sensitivity analysis found peak year budget impact varying between 2% and 11%. The budget impact of OMPs in Sweden and France is likely to remain sustainable over time and a relatively small proportion of total pharmaceutical expenditure. This forecast could be affected by changes in the success rate for OMP approvals, average cost of OMPs, and the type of companies developing OMPs.

Bacteroides Fragilis OmpA: Utility as a Live Vaccine Vector for Biodefense Agents

DTIC Science & Technology

2009-01-01

shared by all four homologs are highlighted in black. Positions where amino acid similarity is shared by all four homologs are highlighted in gray ...study Primers for RT PCR OmpA1-F gga tat gac ggt gtt gcc ag this study OmpA1-R tag cag cag cca tgt cat tc this study OmpA2-F tag aag gtg cat...gga cta ct this study OmpA2-R aac cgc caa tag cat tgg ac this study OmpA3-F act ccg ctg atc aat gtg tc this study OmpA3-R cgt ctg cac gca tag tga ag

Assessment of Orbital-Optimized Third-Order Møller-Plesset Perturbation Theory and Its Spin-Component and Spin-Opposite Scaled Variants for Thermochemistry and Kinetics.

PubMed

Soydaş, Emine; Bozkaya, Uğur

2013-03-12

An assessment of the OMP3 method and its spin-component and spin-scaled variants for thermochemistry and kinetics is presented. For reaction energies of closed-shell systems, the CCSD, SCS-MP3, and SCS-OMP3 methods show better performances than other considered methods, and no significant improvement is observed due to orbital optimization. For barrier heights, OMP3 and SCS-OMP3 provide the lowest mean absolute deviations. The MP3 method yields considerably higher errors, and the spin scaling approaches do not help to improve upon MP3, but worsen it. For radical stabilization energies, the CCSD, OMP3, and SCS-OMP3 methods exhibit noticeably better performances than MP3 and its variants. Our results demonstrate that if the reference wave function suffers from a spin-contamination, then the MP3 methods dramatically fail. On the other hand, the OMP3 method and its variants can tolerate the spin-contamination in the reference wave function. For overall evaluation, we conclude that OMP3 is quite helpful, especially in electronically challenged systems, such as free radicals or transition states where spin contamination dramatically deteriorates the quality of the canonical MP3 and SCS-MP3 methods. Both OMP3 and CCSD methods scale as n(6), where n is the number of basis functions. However, the OMP3 method generally converges in much fewer iterations than CCSD. In practice, OMP3 is several times faster than CCSD in energy computations. Further, the stationary properties of OMP3 make it much more favorable than CCSD in the evaluation of analytic derivatives. For OMP3, the analytic gradient computations are much less expensive than CCSD. For the frequency computation, both methods require the evaluation of the perturbed amplitudes and orbitals. However, in the OMP3 case there is still a significant computational time savings due to simplifications in the analytic Hessian expression owing to the stationary property of OMP3. Hence, the OMP3 method emerges as a very useful tool for computational quantum chemistry.

Principles for consistent value assessment and sustainable funding of orphan drugs in Europe.

PubMed

Gutierrez, Laura; Patris, Julien; Hutchings, Adam; Cowell, Warren

2015-05-03

The European Orphan Medicinal Products (OMP) Regulation has successfully encouraged research to develop treatments for rare diseases resulting in the authorisation of new OMPs in Europe. While decisions on OMP designation and marketing authorisation are made at the European Union level, reimbursement decisions are made at the national level. OMP value and affordability are high priority issues for policymakers and decisions regarding their pricing and funding are highly complex. There is currently no European consensus on how OMP value should be assessed and inequalities of access to OMPs have previously been observed. Against this background, policy makers in many countries are considering reforms to improve access to OMPs. This paper proposes ten principles to be considered when undertaking such reforms, from the perspective of an OMP manufacturer. We recommend the continued prioritisation of rare diseases by policymakers, an increased alignment between payer and regulatory frameworks, pricing centred on OMP value, and mechanisms to ensure long-term financial sustainability allowing a continuous and virtuous development of OMPs. Our recommendations support the development of more consistent frameworks and encourage collaboration between all stakeholders, including research-based industry, payers, clinicians, and patients.

Transcutaneous immunization with an outer membrane protein of Porphyromonas gingivalis without adjuvant elicits marked antibody responses.

PubMed

Koizumi, Y; Kurita-Ochiai, T; Yamamoto, M

2008-04-01

We have previously reported that specific immunoglobulin G (IgG) antibodies induced by transcutaneous immunization (TCI) with a 40-kDa outer membrane protein (40k-OMP) of Porphyromonas gingivalis, with cholera toxin (CT) as adjuvant, inhibited coaggregation by P. gingivalis. In this study, we further pursue the potential of the 40k-OMP as a transcutaneous vaccine. TCI of rats administered 40k-OMP elicited significant 40k-OMP-specific serum IgG and IgA, as well as salivary IgG antibody titers. Importantly, these antibody responses were induced without adjuvant. Thus, both serum and saliva antibody titers induced by TCI with the 40k-OMP alone were identical to those of 40k-OMP plus cholera toxin as adjuvant. The serum antibody responses induced by 40k-OMP persisted for more than 140 days. On the other hand, salivary IgG anti-40k-OMP antibodies were gradually decreased. Analysis of antibody-forming cells (AFCs) confirmed the antibody titers by detecting high numbers of 40k-OMP-specific IgG AFCs in spleen and cervical lymph node. Since 40k-OMP-specific IgG inhibited the coaggregation of P. gingivalis with Streptococcus gordonii, and the hemagglutinin activity of P. gingivalis, TCI with the 40k-OMP may be important as an adjuvant-free immunogen for the prevention of chronic periodontitis.

Biochemical and functional characterization of the periplasmic domain of the outer membrane protein A from enterohemorrhagic Escherichia coli.

PubMed

Wang, Haiguang; Li, Qian; Fang, Yao; Yu, Shu; Tang, Bin; Na, Li; Yu, Bo; Zou, Quanming; Mao, Xuhu; Gu, Jiang

2016-01-01

Outer membrane protein A (OmpA) plays multiple roles in the physiology and pathogenesis of the zoonotic pathogen enterohemorrhagic Escherichia coli (EHEC). The N-terminus of OmpA forms a transmembrane domain (OmpA™), and the roles of this domain in bacterial pathogenesis have been well studied. However, how its C-terminal domain (OmpAper), which is located at the periplasmic space in the bacterial membrane, contributes to virulence remains unclear. Herein, we report that OmpAper forms a dimer and binds to peptidoglycan in vitro. Furthermore, OmpAper is responsible for bacterial resistance to acidic conditions, high osmotic pressure and high SDS environments. In addition, OmpAper contributes to the adhesion of bacteria to HeLa cells in vitro and ex vivo. These results provide an additional understanding of the role of OmpA in EHEC physiology and pathogenesis. Copyright © 2015 Elsevier GmbH. All rights reserved.

Rational Design and Evaluation of an Artificial Escherichia coli K1 Protein Vaccine Candidate Based on the Structure of OmpA

PubMed Central

Gu, Hao; Liao, Yaling; Zhang, Jin; Wang, Ying; Liu, Zhiyong; Cheng, Ping; Wang, Xingyong; Zou, Quanming; Gu, Jiang

2018-01-01

Escherichia coli (E. coli) K1 causes meningitis and remains an unsolved problem in neonates, despite the application of antibiotics and supportive care. The cross-reactivity of bacterial capsular polysaccharides with human antigens hinders their application as vaccine candidates. Thus, protein antigens could be an alternative strategy for the development of an E. coli K1 vaccine. Outer membrane protein A (OmpA) of E. coli K1 is a potential vaccine candidate because of its predominant contribution to bacterial pathogenesis and sub-cellular localization. However, little progress has been made regarding the use of OmpA for this purpose due to difficulties in OmpA production. In the present study, we first investigated the immunogenicity of the four extracellular loops of OmpA. Using the structure of OmpA, we rationally designed and successfully generated the artificial protein OmpAVac, composed of connected loops from OmpA. Recombinant OmpAVac was successfully produced in E. coli BL21 and behaved as a soluble homogenous monomer in the aqueous phase. Vaccination with OmpAVac induced Th1, Th2, and Th17 immune responses and conferred effective protection in mice. In addition, OmpAVac-specific antibodies were able to mediate opsonophagocytosis and inhibit bacterial invasion, thereby conferring prophylactic protection in E. coli K1-challenged adult mice and neonatal mice. These results suggest that OmpAVac could be a good vaccine candidate for the control of E. coli K1 infection and provide an additional example of structure-based vaccine design. PMID:29876324

Rational Design and Evaluation of an Artificial Escherichia coli K1 Protein Vaccine Candidate Based on the Structure of OmpA.

PubMed

Gu, Hao; Liao, Yaling; Zhang, Jin; Wang, Ying; Liu, Zhiyong; Cheng, Ping; Wang, Xingyong; Zou, Quanming; Gu, Jiang

2018-01-01

Escherichia coli ( E. coli ) K1 causes meningitis and remains an unsolved problem in neonates, despite the application of antibiotics and supportive care. The cross-reactivity of bacterial capsular polysaccharides with human antigens hinders their application as vaccine candidates. Thus, protein antigens could be an alternative strategy for the development of an E. coli K1 vaccine. Outer membrane protein A (OmpA) of E. coli K1 is a potential vaccine candidate because of its predominant contribution to bacterial pathogenesis and sub-cellular localization. However, little progress has been made regarding the use of OmpA for this purpose due to difficulties in OmpA production. In the present study, we first investigated the immunogenicity of the four extracellular loops of OmpA. Using the structure of OmpA, we rationally designed and successfully generated the artificial protein OmpAVac, composed of connected loops from OmpA. Recombinant OmpAVac was successfully produced in E. coli BL21 and behaved as a soluble homogenous monomer in the aqueous phase. Vaccination with OmpAVac induced Th1, Th2, and Th17 immune responses and conferred effective protection in mice. In addition, OmpAVac-specific antibodies were able to mediate opsonophagocytosis and inhibit bacterial invasion, thereby conferring prophylactic protection in E. coli K1-challenged adult mice and neonatal mice. These results suggest that OmpAVac could be a good vaccine candidate for the control of E. coli K1 infection and provide an additional example of structure-based vaccine design.

The surface protease ompT serves as Escherichia coli K1 adhesin in binding to human brain micro vascular endothelial cells.

PubMed

Wan, Lei; Guo, Yan; Hui, Chang-Ye; Liu, Xiao-Lu; Zhang, Wen-Bing; Cao, Hong; Cao, Hong

2014-05-01

Escherichia coli (E. coli) K1 is the most common bacteria that cause meningitis in the neonatal period. But it's not entirely clear about how E. coli crosses the blood-brain barrier. The features of the ompT deletion in meningitic E. coli infection were texted in vitro. In comparison with the parent strain, the isogenic ompT deletion mutant was significantly less adhesive to human brain microvascular endothelial cells (HBMEC). The adhesion-deficient phenotype of the mutant was restored to the level of the wild-type by complementing with low-level OmpT expression plasmid. Interestingly, the adhesion was enhanced by point mutation at the OmpT proposed catalytic residue D85. Compared with the poor adhesive activity of bovine serum albumin-coated fluorescent beads, recombinant OmpT or catalytically inactive variant of OmpT-coated beads bound to HBMEC monolayer effectively. Our study suggests that OmpT is important for bacterial adhesion while entering into central nervous system, and the adhesion does not involve in the proteolytic activity of OmpT.

Chlamydia trachomatis OmpA genotyping as a tool for studying the natural history of genital chlamydial infection.

PubMed

Geisler, W M; Black, C M; Bandea, C I; Morrison, S G

2008-12-01

To investigate the relationship of Chlamydia trachomatis (CT) outer membrane protein A (OmpA) type to the clearance of CT infection before treatment. CT OmpA genotyping, with amplification and sequencing of ompA, was utilised to study the natural history of CT infection (spontaneous resolution vs persistence) in 102 individuals with chlamydia-positive screening tests returning for treatment. CT OmpA distribution was associated with spontaneous resolution of CT, most notably with CT OmpA genotype J/Ja detected more often from the initial screening CT test than other genotypes in those who then had spontaneous resolution of CT noted at the time of treatment. Five individuals with presumed persisting CT infection had discordant CT OmpA genotypes at the screening and treatment visits, suggesting possible new interval CT infection. Clearance of chlamydia by the host before treatment may be influenced by the CT OmpA genotype infecting the host. CT OmpA genotyping may be a valuable tool in understanding the natural history of chlamydial infections.

Molecular characterization and histochemical demonstration of salmon olfactory marker protein in the olfactory epithelium of lacustrine sockeye salmon (Oncorhynchus nerka).

PubMed

Kudo, H; Doi, Y; Ueda, H; Kaeriyama, M

2009-09-01

Despite the importance of olfactory receptor neurons (ORNs) for homing migration, the expression of olfactory marker protein (OMP) is not well understood in ORNs of Pacific salmon (genus Oncorhynchus). In this study, salmon OMP was characterized in the olfactory epithelia of lacustrine sockeye salmon (O. nerka) by molecular biological and histochemical techniques. Two cDNAs encoding salmon OMP were isolated and sequenced. These cDNAs both contained a coding region encoding 173 amino acid residues, and the molecular mass of the two proteins was calculated to be 19,581.17 and 19,387.11Da, respectively. Both amino acid sequences showed marked homology (90%). The protein and nucleotide sequencing demonstrates the existence of high-level homology between salmon OMPs and those of other teleosts. By in situ hybridization using a digoxigenin-labeled salmon OMP cRNA probe, signals for salmon OMP mRNA were observed preferentially in the perinuclear regions of the ORNs. By immunohistochemistry using a specific antibody to salmon OMP, OMP-immunoreactivities were noted in the cytosol of those neurons. The present study is the first to describe cDNA cloning of OMP in salmon olfactory epithelium, and indicate that OMP is a useful molecular marker for the detection of the ORNs in Pacific salmon.

Structural analysis and cross-protective efficacy of recombinant 87 kDa outer membrane protein (Omp87) of Pasteurella multocida serogroup B:2.

PubMed

Kumar, Abhinendra; Yogisharadhya, Revanaiah; Ramakrishnan, Muthannan A; Viswas, K N; Shivachandra, Sathish B

2013-12-01

Pasteurella multocida serogroup B:2, a causative agent of haemorrhagic septicaemia (HS) in cattle and buffalo especially in tropical regions of Asian and African countries, is known to possess several outer membrane proteins (OMPs) as immunogenic antigens. In the present study, omp87 gene encoding for 87 kDa OMP (Omp87) protein of P. multocida serogroup B:2 strain P52, has been amplified (∼2304 bp), cloned in to pET32a vector and over-expressed in recombinant Escherichia coli as fusion protein. The recombinant Omp87 protein (∼102 kDa) including N-terminus hexa-histidine tag was purified under denaturing condition. Immunization of mice with rOmp87 resulted in increased antigen specific IgG titres in serum and provided protection of 66.6 and 83.3% following homologous (B:2) and heterologous (A:1) challenge, respectively. A homology model of Omp87 revealed the presence of two distinct domains; N-terminal domain with four POTRA repeats in the periplasmic space and a pore forming C-terminal β-barrel domain (β1- β16) in the outer membrane of P. multocida, which belong to Omp85-TpsB transporter superfamily of OMPs. The study indicated the potential possibilities to use rOmp87 protein along with suitable adjuvant in developing subunit vaccine for haemorrhagic septicaemia and pasteurellosis in livestock. Copyright © 2013 Elsevier Ltd. All rights reserved.

Granular activated carbon adsorption of organic micro-pollutants in drinking water and treated wastewater--Aligning breakthrough curves and capacities.

PubMed

Zietzschmann, Frederik; Stützer, Christian; Jekel, Martin

2016-04-01

Small-scale granular activated carbon (GAC) tests for the adsorption of organic micro-pollutants (OMP) were conducted with drinking water and wastewater treatment plant (WWTP) effluent. In both waters, three influent OMP concentration levels were tested. As long as the influent OMP concentrations are below certain thresholds, the relative breakthrough behavior is not impacted in the respective water. Accordingly, the GAC capacity for OMP is directly proportional to the influent OMP concentration in the corresponding water. The differences between the OMP breakthrough curves in drinking water and WWTP effluent can be attributed to the concentrations of the low molecular weight acid and neutral (LMW) organics of the waters. Presenting the relative OMP concentrations (c/c0) over the specific throughput of the LMW organics (mg LMW organics/g GAC), the OMP breakthrough curves in drinking water and WWTP effluent superimpose each other. This superimposition can be further increased if the UV absorbance at 254 nm (UV254) of the LMW organics is considered. In contrast, using the specific throughput of the dissolved organic carbon (DOC) did not suffice to obtain superimposed breakthrough curves. Thus, the LMW organics are the major water constituent impacting OMP adsorption onto GAC. The results demonstrate that knowing the influent OMP and LMW organics concentrations (and UV254) of different waters, the OMP breakthroughs and GAC capacities corresponding to any water can be applied to all other waters. Copyright © 2016 Elsevier Ltd. All rights reserved.

The role of systemic steroids and phototherapy in the treatment of stable vitiligo: a randomized controlled trial.

PubMed

El Mofty, Medhat; Essmat, Samia; Youssef, Randa; Sobeih, Sherine; Mahgoub, Doaa; Ossama, Sherine; Saad, Akmal; El Tawdy, Amira; Mashaly, Heba M; Saney, Iman; Helal, Rana; Shaker, Olfat

2016-11-01

Pathogenesis of vitiligo is believed to be multifactorial disease with a wide variety of therapeutic modalities. The aim of this work is to assess the efficacy of oral mini-pulse steroids (OMP) plus Nb-U.V.B in comparison to OMP alone and Nb-U.V.B alone in treating stable vitiligo. A prospective randomized controlled study including 45 patients categorized into three groups receiving therapy for 3 months; Group A received Nb-U.V.B plus OMP, Group B received OMP alone while Group C received Nb-U.V.B alone. Clinical assessment and PCR evaluation of bFGF, ICAM1, and ELISA for AMA were done. Patients receiving Nb-U.V.B plus OMP and using Nb-U.V.B alone gave statistically significant clinical response than those treated with OMP alone. Statistically significant rise of BFGF was noticed after treatment with Nb-U.V.B plus OMP and with Nb-U.V.B alone. Patients treated with OMP alone and with Nb-U.V.B alone showed statistically significant drop of ICAM-1 after therapy. NB-U.V.B plus OMP and Nb-U.V.B alone were found to be clinically superior over OMP alone in treating stable vitiligo patients, hence suggesting that adding OMP to Nb-U.V.B can maintain clinical and laboratory success for a longer period of time and with less relapse. © 2016 Wiley Periodicals, Inc.

Klebsiella pneumoniae Outer Membrane Protein A Is Required to Prevent the Activation of Airway Epithelial Cells*

PubMed Central

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

2011-01-01

Outer membrane protein A (OmpA) is a class of proteins highly conserved among the Enterobacteriaceae family and throughout evolution. Klebsiella pneumoniae is a capsulated Gram-negative pathogen. It is an important cause of community-acquired and nosocomial pneumonia. Evidence indicates that K. pneumoniae infections are characterized by a lack of an early inflammatory response. Data from our laboratory indicate that K. pneumoniae CPS helps to suppress the host inflammatory response. However, it is unknown whether K. pneumoniae employs additional factors to modulate host inflammatory responses. Here, we report that K. pneumoniae OmpA is important for immune evasion in vitro and in vivo. Infection of A549 and normal human bronchial cells with 52OmpA2, an ompA mutant, increased the levels of IL-8. 52145-ΔwcaK2ompA, which does not express CPS and ompA, induced the highest levels of IL-8. Both mutants could be complemented. In vivo, 52OmpA2 induced higher levels of tnfα, kc, and il6 than the wild type. ompA mutants activated NF-κB, and the phosphorylation of p38, p44/42, and JNK MAPKs and IL-8 induction was via NF-κB-dependent and p38- and p44/42-dependent pathways. 52OmpA2 engaged TLR2 and -4 to activate NF-κB, whereas 52145-ΔwcaK2ompA activated not only TLR2 and TLR4 but also NOD1. Finally, we demonstrate that the ompA mutant is attenuated in the pneumonia mouse model. The results of this study indicate that K. pneumoniae OmpA contributes to attenuate airway cell responses. This may facilitate pathogen survival in the hostile environment of the lung. PMID:21278256

Molecular identification of the ompL1 gene within Leptospira interrogans standard serovars.

PubMed

Dezhbord, Mehrangiz; Esmaelizad, Majid; Khaki, Pejvak; Fotohi, Fariba; Zarehparvar Moghaddam, Athena

2014-06-11

Leptospirosis, caused by infection with pathogenic Leptospira species, is one of the most prevalent zoonotic diseases in the world. Current leptospiral vaccines are mainly multivalent dead whole-cell mixtures made of several local dominant serovars. Therefore, design and construction of an efficient recombinant vaccine for leptospirosis control is very important. OmpL1 is an immunogenic porin protein that could be of special significance in vaccination and serodiagnosis for leptospirosis. Three strains belonging to pathogenic L. interrogans were analyzed. The specific primers for proliferation of the ompL1 gene were designed. The amplified gene was cloned. In order to investigate the ompL1 nucleotide sequence and homological analysis of this gene, ompL1 genes cloned from standard vaccinal Leptospira serovars prevalent in Iran were sequenced and cloned. PCR amplification of the ompL1 gene using the designed primers resulted in a 963 bp ompL1 gene product. The PCR based on the ompL1 gene detected all pathogenic reference serovars of Leptospira spp. tested. Based on alignment and phylogenetic analysis, although the ompL1 nucleotide sequence was slightly different within three vaccinal serovars (100%-85% identity), amino acid alignment of the OmpL1 proteins revealed that there would be inconsiderable difference among them. The ompL1 gene of the three isolates was well conserved, differing only by a total of 6 bp and the proteins by 2 amino acids. The cloned gene could be further used for expression and recombinant OmpL1 as an efficient and conserved antigen, and may be a useful vaccine candidate against leptospirosis in our region.

Regulation of Serratia marcescens ompF and ompC porin genes in response to osmotic stress, salicylate, temperature and pH.

PubMed

Begic, Sanela; Worobec, Elizabeth A

2006-02-01

Serratia marcescens is a Gram-negative enterobacterium that has become an important opportunistic pathogen, largely due to its high degree of natural antibiotic resistance. One factor contributing to this natural antibiotic resistance is reduced outer membrane permeability, which is controlled in part by OmpC and OmpF porin proteins. OmpF expression is regulated by micF, an RNA transcript encoded upstream of the ompC gene, which hybridizes with the ompF transcript to inhibit its translation. Regulation of S. marcescens porin gene expression, as well as that of micF, was investigated using beta-galactosidase reporter gene fusions in response to 5, 8 and 10 % sucrose, 1, 5 and 8 mM salicylate, and different pH and temperature values. beta-Galactosidase activity assays revealed that a lower growth temperature (28 degrees C), a more basic pH (pH 8), and an absence of sucrose and salicylate induce the transcription of the ompF gene, whereas the induction of ompC is stimulated at a higher growth temperature (42 degrees C), acidic pH (pH 6), and maximum concentrations of sucrose (10 %) and salicylate (8 mM). In addition, when multiple conditions were tested, temperature had the predominant effect, followed by pH. In this study, it was found that the MicF regulatory mechanism does not play a role in the osmoregulation of the ompF and ompC genes, whereas MicF does repress OmpF expression in the presence of salicylate and high growth temperature, and under low pH conditions.

CdiA Effectors from Uropathogenic Escherichia coli Use Heterotrimeric Osmoporins as Receptors to Recognize Target Bacteria

PubMed Central

Beck, Christina M.; Willett, Julia L. E.; Kim, Jeff J.; Low, David A.; Hayes, Christopher S.

2016-01-01

Many Gram-negative bacterial pathogens express contact-dependent growth inhibition (CDI) systems that promote cell-cell interaction. CDI+ bacteria express surface CdiA effector proteins, which transfer their C-terminal toxin domains into susceptible target cells upon binding to specific receptors. CDI+ cells also produce immunity proteins that neutralize the toxin domains delivered from neighboring siblings. Here, we show that CdiAEC536 from uropathogenic Escherichia coli 536 (EC536) uses OmpC and OmpF as receptors to recognize target bacteria. E. coli mutants lacking either ompF or ompC are resistant to CDIEC536-mediated growth inhibition, and both porins are required for target-cell adhesion to inhibitors that express CdiAEC536. Experiments with single-chain OmpF fusions indicate that the CdiAEC536 receptor is heterotrimeric OmpC-OmpF. Because the OmpC and OmpF porins are under selective pressure from bacteriophages and host immune systems, their surface-exposed loops vary between E. coli isolates. OmpC polymorphism has a significant impact on CDIEC536 mediated competition, with many E. coli isolates expressing alleles that are not recognized by CdiAEC536. Analyses of recombinant OmpC chimeras suggest that extracellular loops L4 and L5 are important recognition epitopes for CdiAEC536. Loops L4 and L5 also account for much of the sequence variability between E. coli OmpC proteins, raising the possibility that CDI contributes to the selective pressure driving OmpC diversification. We find that the most efficient CdiAEC536 receptors are encoded by isolates that carry the same cdi gene cluster as E. coli 536. Thus, it appears that CdiA effectors often bind preferentially to "self" receptors, thereby promoting interactions between sibling cells. As a consequence, these effector proteins cannot recognize nor suppress the growth of many potential competitors. These findings suggest that self-recognition and kin selection are important functions of CDI. PMID:27723824

Site-directed mutagenesis studies to probe the role of specific residues in the external loop (L3) of OmpF and OmpC porins in susceptibility of Serratia marcescens to antibiotics.

PubMed

Begic, Sanela; Worobec, Elizabeth A

2007-06-01

Serratia marcescens is a nosocomial bacterium with natural resistance to a broad spectrum of antibiotics, making treatment challenging. One factor contributing to this natural antibiotic resistance is reduced outer membrane permeability, controlled in part by OmpF and OmpC porin proteins. To investigate the direct role of these porins in the diffusion of antibiotics across the outer membrane, we have created an ompF-ompC porin-deficient strain of S. marcescens. A considerable similarity between the S. marcescens porins and those from other members of Enterobacteriaceae was detected by sequence alignment, with the exception of a change in a conserved region of the third external loop (L3) of the S. marcescens OmpC protein. Serratia marcescens OmpC has aspartic acid instead of glycine in position 112, methionine instead of aspartic acid in position 114, and glutamine in position 124, while in S. marcescens OmpF this is a glycine at position 124. To investigate the role of amino acid positions 112, 114, and 124 and how the observed changes within OmpC porin may play a part in pore permeability, 2 OmpC sites were altered in the Enterobacteriaceae consensus (D112G and M114D) through site-directed mutagenesis. Also, Q124G in OmpC, G124Q in OmpF, and double mutants of these amino acid residues were constructed. Antibiotic accumulation assays and minimal inhibitory concentrations of the strains harboring the mutated porins were performed, while liposome swelling experiments were performed on purified porins. Our results demonstrate that the amino acid at position 114 is not responsible for either antibiotic size or ionic selection, the amino acid at position 112 is responsible for size selection only, and position 124 is involved in both size and ionic selection.

The Role of OmpK35, OmpK36 Porins, and Production of β-Lactamases on Imipenem Susceptibility in Klebsiella pneumoniae Clinical Isolates, Cairo, Egypt.

PubMed

Wassef, Mona; Abdelhaleim, Mona; AbdulRahman, Eiman; Ghaith, Doaa

2015-12-01

OmpK35 and OmpK36 are the major outer membrane porins of Klebsiella pneumoniae. We aimed to study the effect of combined porin loss and production of extended-spectrum β-lactamases (ESBLs) on imipenem susceptibility among K. pneumoniae clinical isolates. This study included 91 suspected ESBL-producing K. pneumoniae clinical isolates, isolated from different patient specimens at the Cairo University hospital from January to June 2010. All isolates were subjected to genotypic analysis of the outer membrane protein gene expression using reverse transcription-PCR (RT-PCR) and analysis of OmpK35/36 of 38 isolates by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). By RT-PCR, loss of Omp35 was detected in 78 (85.7%) isolates, loss of Omp36 was detected in 64 (70.32%), and loss of both porins was detected in 62 (68.1%). Out of 91 isolates, 45 (49.5%) were resistant to cefoxitin, and 17 (18.7%) were confirmed as derepressed AmpC producers. Omp35 was lost in all FOX-resistant isolates, whereas Omp36 was lost in 42 (93.3%) (p-value 0.002). The mean of ceftazidime inhibition zone diameter was significantly decreased among ESBL-producing isolates with loss of Omp35/36 (p-value 0.041 and 0.006), respectively. The mean of imipenem minimal inhibitory concentration (MIC) was markedly increased to 8.55 μg/ml among AmpC-producing isolates with Omp35/36 loss, while the mean of imipenem MIC among the 66 confirmed ESBL producers was 0.32 μg/ml. Imipenem MIC was markedly increased among K. pneumoniae isolates showing AmpC production with loss of both porins OmpK35/36. Meanwhile, the association of porin OmpK35/36 loss with ESBL production was not a direct cause of resistance to imipenem.

Comparative proteome analysis reveals pathogen specific outer membrane proteins of Leptospira.

PubMed

Dhandapani, Gunasekaran; Sikha, Thoduvayil; Rana, Aarti; Brahma, Rahul; Akhter, Yusuf; Gopalakrishnan Madanan, Madathiparambil

2018-04-10

Proteomes of pathogenic Leptospira interrogans and L. borgpetersenii and the saprophytic L. biflexa were filtered through computational tools to identify Outer Membrane Proteins (OMPs) that satisfy the required biophysical parameters for their presence on the outer membrane. A total of 133, 130, and 144 OMPs were identified in L. interrogans, L. borgpetersenii, and L. biflexa, respectively, which forms approximately 4% of proteomes. A holistic analysis of transporting and pathogenic characteristics of OMPs together with Clusters of Orthologous Groups (COGs) among the OMPs and their distribution across 3 species was made and put forward a set of 21 candidate OMPs specific to pathogenic leptospires. It is also found that proteins homologous to the candidate OMPs were also present in other pathogenic species of leptospires. Six OMPs from L. interrogans and 2 from L. borgpetersenii observed to have similar COGs while those were not found in any intermediate or saprophytic forms. These OMPs appears to have role in infection and pathogenesis and useful for anti-leptospiral strategies. © 2018 Wiley Periodicals, Inc.

Homology analysis and cross-immunogenicity of OmpA from pathogenic Yersinia enterocolitica, Yersinia pseudotuberculosis and Yersinia pestis.

PubMed

Chen, Yuhuang; Duan, Ran; Li, Xu; Li, Kewei; Liang, Junrong; Liu, Chang; Qiu, Haiyan; Xiao, Yuchun; Jing, Huaiqi; Wang, Xin

2015-12-01

The outer membrane protein A (OmpA) is one of the intra-species conserved proteins with immunogenicity widely found in the family of Enterobacteriaceae. Here we first confirmed OmpA is conserved in the three pathogenic Yersinia: Yersinia pestis, Yersinia pseudotuberculosis and pathogenic Yersinia enterocolitica, with high homology at the nucleotide level and at the amino acid sequence level. The identity of ompA sequences for 262 Y. pestis strains, 134 Y. pseudotuberculosis strains and 219 pathogenic Y. enterocolitica strains are 100%, 98.8% and 97.7% similar. The main pattern of OmpA of pathogenic Yersinia are 86.2% and 88.8% identical at the nucleotide and amino acid sequence levels, respectively. Immunological analysis showed the immunogenicity of each OmpA and cross-immunogenicity of OmpA for pathogenic Yersinia where OmpA may be a vaccine candidate for Y. pestis and other pathogenic Yersinia. Copyright © 2015 Elsevier Ltd. All rights reserved.

Antibody response of swine to outer membrane components of Haemophilus pleuropneumoniae during infection.

PubMed Central

Rapp, V J; Ross, R F

1986-01-01

Sera from pigs infected with Haemophilus (Actinobacillus) pleuropneumoniae were tested for antibodies to outer membrane proteins (OMPs) of the organism by immunoblotting. Convalescent sera were produced in naturally born, colostrum-fed pigs and in cesarean-derived, colostrum-deprived pigs given H. pleuropneumoniae serotype 5 intranasally twice at 5-week intervals. Sera, collected at weekly intervals, were reacted with Sarkosyl-insoluble, OMP-enriched preparations of H. pleuropneumoniae which had been separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electrophoretically transferred to nitrocellulose. Antibodies were detected to OMPs with an apparent molecular weight of 16,500 (16.5K OMP); to 29K, 38.5K, 43.5K, 45K, 49.5K, and 66.5K OMPs; and to several high-molecular-weight (greater than or equal to 94,000) OMPs, but not to the major 42K OMP. Antibodies to the heat-modifiable OMP (29K/43.5K) and the 38.5K OMP were detected in sera from noninfected pigs. Antibodies were also detected to two broad 54,000- and 95,000-molecular-weight bands which did not stain with Coomassie blue, stained with silver nitrate, resisted proteinase K digestion, and were eliminated by oxidation with sodium metaperiodate. This indicates that the 54,000- and 95,000-molecular-weight bands represent polysaccharide, possibly capsular or lipopolysaccharide immunogens. Adsorption of sera with cells from the homologous serotype 5 strain removed antibodies to the 45K, 49.5K, 66.5K, and greater than or equal to 94K OMPs and to the two polysaccharide bands, indicating that these antibodies were directed primarily to surface-exposed epitopes. When tested with OMP preparations from other serotype 5 strains, heterogeneity was apparent, both in the reactions with OMPs and with the polysaccharide bands. Silver staining of proteinase K-treated, whole-cell lysates from serotype 5 strains also indicated variable expression of the polysaccharide bands. Sera also reacted with OMPs from H. pleuropneumoniae serotypes 1 and 7; however, several OMPs and the lipopolysaccharide or polysaccharide determinants of these serotypes appeared to be type specific. Images PMID:3536748

Assessment of Orbital-Optimized MP2.5 for Thermochemistry and Kinetics: Dramatic Failures of Standard Perturbation Theory Approaches for Aromatic Bond Dissociation Energies and Barrier Heights of Radical Reactions.

PubMed

Soydaş, Emine; Bozkaya, Uğur

2015-04-14

An assessment of orbital-optimized MP2.5 (OMP2.5) [ Bozkaya, U.; Sherrill, C. D. J. Chem. Phys. 2014, 141, 204105 ] for thermochemistry and kinetics is presented. The OMP2.5 method is applied to closed- and open-shell reaction energies, barrier heights, and aromatic bond dissociation energies. The performance of OMP2.5 is compared with that of the MP2, OMP2, MP2.5, MP3, OMP3, CCSD, and CCSD(T) methods. For most of the test sets, the OMP2.5 method performs better than MP2.5 and CCSD, and provides accurate results. For barrier heights of radical reactions and aromatic bond dissociation energies OMP2.5-MP2.5, OMP2-MP2, and OMP3-MP3 differences become obvious. Especially, for aromatic bond dissociation energies, standard perturbation theory (MP) approaches dramatically fail, providing mean absolute errors (MAEs) of 22.5 (MP2), 17.7 (MP2.5), and 12.8 (MP3) kcal mol(-1), while the MAE values of the orbital-optimized counterparts are 2.7, 2.4, and 2.4 kcal mol(-1), respectively. Hence, there are 5-8-folds reductions in errors when optimized orbitals are employed. Our results demonstrate that standard MP approaches dramatically fail when the reference wave function suffers from the spin-contamination problem. On the other hand, the OMP2.5 method can reduce spin-contamination in the unrestricted Hartree-Fock (UHF) initial guess orbitals. For overall evaluation, we conclude that the OMP2.5 method is very helpful not only for challenging open-shell systems and transition-states but also for closed-shell molecules. Hence, one may prefer OMP2.5 over MP2.5 and CCSD as an O(N(6)) method, where N is the number of basis functions, for thermochemistry and kinetics. The cost of the OMP2.5 method is comparable with that of CCSD for energy computations. However, for analytic gradient computations, the OMP2.5 method is only half as expensive as CCSD.

Analysis and Characterization of Proteins Associated with Outer Membrane Vesicles Secreted by Cronobacter spp.

PubMed Central

Kothary, Mahendra H.; Gopinath, Gopal R.; Gangiredla, Jayanthi; Rallabhandi, Prasad V.; Harrison, Lisa M.; Yan, Qiong Q.; Chase, Hannah R.; Lee, Boram; Park, Eunbi; Yoo, YeonJoo; Chung, Taejung; Finkelstein, Samantha B.; Negrete, Flavia J.; Patel, Isha R.; Carter, Laurenda; Sathyamoorthy, Venugopal; Fanning, Séamus; Tall, Ben D.

2017-01-01

Little is known about secretion of outer membrane vesicles (OMVs) by Cronobacter. In this study, OMVs isolated from Cronobacter sakazakii, Cronobacter turicensis, and Cronobacter malonaticus were examined by electron microscopy (EM) and their associated outer membrane proteins (OMP) and genes were analyzed by SDS-PAGE, protein sequencing, BLAST, PCR, and DNA microarray. EM of stained cells revealed that the OMVs are secreted as pleomorphic micro-vesicles which cascade from the cell's surface. SDS-PAGE analysis identified protein bands with molecular weights of 18 kDa to >100 kDa which had homologies to OMPs such as GroEL; OmpA, C, E, F, and X; MipA proteins; conjugative plasmid transfer protein; and an outer membrane auto-transporter protein (OMATP). PCR analyses showed that most of the OMP genes were present in all seven Cronobacter species while a few genes (OMATP gene, groEL, ompC, mipA, ctp, and ompX) were absent in some phylogenetically-related species. Microarray analysis demonstrated sequence divergence among the OMP genes that was not captured by PCR. These results support previous findings that OmpA and OmpX may be involved in virulence of Cronobacter, and are packaged within secreted OMVs. These results also suggest that other OMV-packaged OMPs may be involved in roles such as stress response, cell wall and plasmid maintenance, and extracellular transport. PMID:28232819

Effects of beta-lactamases and omp mutation on susceptibility to beta-lactam antibiotics in Escherichia coli.

PubMed Central

Hiraoka, M; Okamoto, R; Inoue, M; Mitsuhashi, S

1989-01-01

Four types of beta-lactamases consisting of a penicillinase type I (TEM-1), a penicillinase type II (OXA-1), a cephalosporinase of Citrobacter freundii, and a cephalosporinase of Proteus vulgaris were introduced into Escherichia coli MC4100 and its omp mutants, MH1160 (MC4100 ompR1) and MH760 (MC4100 ompR2), by transformation. Effects of the combination of the omp mutations and these beta-lactamases on the susceptibility of E. coli strains were studied with 15 beta-lactam antibiotics including cephalosporins, cephamycins, penicillins, imipenem, and aztreonam. The ompR1 mutant, MH1160, lacks OmpF and OmpC, and it showed reduced susceptibility to 11 of the 15 beta-lactam agents. The reduction in susceptibility to cefoxitin, moxalactam, and flomoxef was much greater than reduction in susceptibility to the other agents. When the ompR1 mutant produced the cephalosporinase of C. freundii, the susceptibility of the mutant to 12 of the 15 beta-lactam antibiotics decreased. The reduction in susceptibility of MH1160 to 10 of the 12 agents affected by the enzyme was two- to fourfold greater than that observed in MC4100. Such a synergistic effect was also observed with the cephalosporinase of P. vulgaris and ompR1 mutation against six cephalosporins, moxalactam, and aztreonam. Images PMID:2658786

Orbital-optimized third-order Møller-Plesset perturbation theory and its spin-component and spin-opposite scaled variants: Application to symmetry breaking problems

NASA Astrophysics Data System (ADS)

Bozkaya, Uǧur

2011-12-01

In this research, orbital-optimized third-order Møller-Plesset perturbation theory (OMP3) and its spin-component and spin-opposite scaled variants (SCS-OMP3 and SOS-OMP3) are introduced. Using a Lagrangian-based approach, an efficient, quadratically convergent algorithm for variational optimization of the molecular orbitals (MOs) for third-order Møller-Plesset perturbation theory (MP3) is presented. Explicit equations for response density matrices, the MO gradient, and Hessian are reported in spin-orbital form. The OMP3, SCS-OMP3, and SOS-OMP3 approaches are compared with the second-order Møller-Plesset perturbation theory (MP2), MP3, coupled-cluster doubles (CCD), optimized-doubles (OD), and coupled-cluster singles and doubles (CCSD) methods. All these methods are applied to the O4 +, O3, and seven diatomic molecules. Results demonstrate that the OMP3 and its variants provide significantly better vibrational frequencies than MP3, CCSD, and OD for the molecules where the symmetry-breaking problems are observed. For O4 +, the OMP3 prediction, 1343 cm-1, for ω6 (b3u) mode, where symmetry-breaking appears, is even better than presumably more reliable methods such as Brueckner doubles (BD), 1194 cm-1, and OD, 1193 cm-1, methods (the experimental value is 1320 cm-1). For O3, the predictions of SCS-OMP3 (1143 cm-1) and SOS-OMP3 (1165 cm-1) are remarkably better than the more robust OD method (1282 cm-1); the experimental value is 1089 cm-1. For the seven diatomics, again the SCS-OMP3 and SOS-OMP3 methods provide the lowest average errors, |Δωe| = 44 and |Δωe| = 35 cm-1, respectively, while for OD, |Δωe| = 161 cm-1and CCSD |Δωe| = 106 cm-1. Hence, the OMP3 and especially its spin-scaled variants perform much better than the MP3, CCSD, and more robust OD approaches for considered test cases. Therefore, considering both the computational cost and the reliability, SCS-OMP3 and SOS-OMP3 appear to be the best methods for the symmetry-breaking cases, based on present application results. The OMP3 method offers certain advantages: it provides reliable vibrational frequencies in case of symmetry-breaking problems, especially with spin-scaling tricks, its analytic gradients are easier to compute since there is no need to solve the coupled-perturbed equations for the orbital response, and the computation of one-electron properties are easier because there is no response contribution to the particle density matrices. The OMP3 has further advantages over standard MP3, making it promising for excited state properties via linear response theory.

Short communication: Roles of outer membrane protein W (OmpW) on survival and biofilm formation of Cronobacter sakazakii under neomycin sulfate stress.

PubMed

Ye, Yingwang; Ling, Na; Gao, Jina; Zhang, Maofeng; Zhang, Xiyan; Tong, Liaowang; Ou, Dexin; Wang, Yaping; Zhang, Jumei; Wu, Qingping

2018-04-01

Cronobacter sakazakii is associated with severe infections including sepsis, neonatal meningitis, and necrotizing enterocolitis. Antibiotic resistance in Cronobacter species has been documented in recent years, but the genes involved in resistance in Cronobacter strains are poorly understood. In this study, we determined the role of outer membrane protein W (OmpW) on survival rates, morphologic changes, and biofilm formation between wild type (WT) and an OmpW mutant strain (ΔOmpW) under neomycin sulfate stress. Results indicated that the survival rates of ΔOmpW were significantly reduced after half minimum inhibitory concentration (½ MIC) treatment compared with the WT strain. Filamentation of C. sakazakii cells was observed after ½ MIC treatment in WT and ΔOmpW, and morphologic injury, including cell disruption and leakage of cells, was more predominant in ΔOmpW. Under ½ MIC stress, the biofilms of WT and ΔOmpW were significantly decreased, but decreasing rates of biofilm formation in mutant strain were more predominant compared with WT strain. This is the first report to determine the role of OmpW on survival, morphological changes, and biofilm formation in C. sakazakii under neomycin sulfate stress. The findings indicated that OmpW contributed to survival and reduction of morphological injury under neomycin sulfate stress. In addition, enhancing biofilm formation in ΔOmpW may be an alternative advantage for adaptation to neomycin sulfate stress. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Florida Public Health Training Center: Evidence-Based Online Mentor Program

ERIC Educational Resources Information Center

Frahm, Kathryn A.; Alsac-Seitz, Biray; Mescia, Nadine; Brown, Lisa M.; Hyer, Kathy; Liburd, Desiree; Rogoff, David P.; Troutman, Adewale

2013-01-01

This article describes an Online Mentor Program (OMP) designed to support and facilitate mentorships among and between Florida Department of Health (FDOH) employees and USF College of Public Health students using a Web-based portal. The Florida Public Health Training Center (FPHTC) at the University of South Florida (USF) College of Public Health…

The role of outer membrane in Serratia marcescens intrinsic resistance to antibiotics.

PubMed

Sánchez, L; Ruiz, N; Leranoz, S; Viñas, M; Puig, M

1997-09-01

Three different porins from Serratia marcescens were described. They were named Omp1, Omp2 and Omp3 and their molecular weights were 42, 40 and 39 kDa respectively. Omp2 and Omp3 showed osmoregulation and thermoregulation in a similar way to OmpC and OmpF of Escherichia coli. Permeability coefficients of the outer membrane of this species were calculated following the Zimmermann and Rosselet method. P values were similar to those obtained in Escherichia coli, which suggests that the chromosomal beta-lactamase would play a major role in the resistance of Serratia marcescens to beta-lactam antibiotics. Both MIC values and permeabilities were modified by salycilates and acetylsalycilate. Synergism between the outer membrane and the beta-lactamase was also evaluated. When bacteria grew in the presence of a beta-lactam in the medium, the beta-lactamase accounted for most of the resistance.

Immunopotentiation of outer membrane protein through anti-idiotype Pasteurella multocida vaccine in rabbits.

PubMed

Arif, Javid; Rahman, Sajjad-Ur; Arshad, Muhammad; Akhtar, Pervez

2013-11-01

Pasteurella multocida was isolated from cattle affected with haemorrhagic septicaemia and characterized on the basis of morphological, cultural and biochemical tests. Bacterial outer membrane proteins (OMPs) were extracted with 1% Sarkosyl method. P. multocida anti-idiotype vaccine prepared from OMPs (21.3 mg per 100 ml), was evaluated and compared with bacterin supplemented with 10% OMPs and plain alum-adsorbed bacterin in rabbit models. It was observed that OMPs-anti-idiotype vaccine induced high levels of antibody titres (geomean titres -GMT) detected using indirect haemagglutination (IHA) test. The OMPs anti-idiotype antibody titres of 168.9 GMT were obtained to 42.2 GMT in OMPs supplemented bacterin on 21 days post vaccination, while the plain bacterin had the least titre of 27.9 GMT. The OMPs-anti-idiotype vaccine provoked better immunogenic response in terms of highest GMT titres and long lasting effect in rabbits and 100% protection against the challenge with homologous strain of P. multocida,while 88% protection was obtained in rabbits, given OMPs supplemented bacterin. Copyright © 2013 The International Alliance for Biological Standardization. Published by Elsevier Ltd. All rights reserved.

Protective immunity induced by the vaccination of recombinant Proteus mirabilis OmpA expressed in Pichia pastoris.

PubMed

Zhang, Yongbing; Yang, Shifa; Dai, Xiumei; Liu, Liping; Jiang, Xiaodong; Shao, Mingxu; Chi, Shanshan; Wang, Chuanwen; Yu, Cuilian; Wei, Kai; Zhu, Ruiliang

2015-01-01

Proteus mirabilis (P. mirabilis) is a zoonotic pathogen that has recently presented a rising infection rate in the poultry industry. To develop an effective vaccine to protect chickens against P. mirabilis infection, OmpA, one of the major outer membrane proteins of P. mirabilis, was expressed in Pichia pastoris. The concentration of the expressed recombinant OmpA protein reached 8.0μg/mL after induction for 96h with 1.0% methanol in the culture. In addition, OmpA protein was confirmed by SDS-PAGE and Western blot analysis using the antibody against Escherichia coli-expressed OmpA protein. Taishan Pinus massoniana pollen polysaccharide, a known plant-derived adjuvant, was mixed into the recombinant OmpA protein to prepare the OmpA subunit vaccine. We then subcutaneously inoculated this vaccine into chickens to examine the immunoprotective effects. ELISA analysis indicated that an excellent antibody response against OmpA was elicited in the vaccinated chickens. Moreover, a high protection rate of 80.0% was observed in the vaccinated group, which was subsequently challenged with P. mirabilis. The results suggest that the eukaryotic P. mirabilis OmpA was an ideal candidate protein for developing an effective subunit vaccine against P. mirabilis infection. Copyright © 2014 Elsevier Inc. All rights reserved.

Comparison of Tropical Ozone from SHADOZ with Remote Sensing Retrievals from Suomi-npp Ozone Mapping Profile Suite (OMPS)

NASA Technical Reports Server (NTRS)

Witte, Jacquelyn C.; Thompson, Anne M.; Ziemke, Jerald R.; Wargan, Krzysztof

2014-01-01

The Ozone Mapping Profile Suite (OMPS) was launched October 28, 2011 on-board the Suomi NPP satellite (http://npp.gsfc.nasa.gov). OMPS is the next generation total column ozone mapping instrument for monitoring the global distribution of stratospheric ozone. OMPS includes a limb profiler to measure the vertical structure of stratosphere ozone down to the mid-troposphere. This study uses tropical ozonesonde profile measurements from the Southern Hemisphere Additional Ozonesondes (SHADOZ, http://croc.gsfc.nasa.gov/shadoz) archive to evaluate total column ozone retrievals from OMPS and concurrent measurements from the Aura Ozone Monitoring Instrument (OMI), the predecessor of OMPS with a data record going back to 2004. We include ten SHADOZ stations that contain data overlapping the OMPS time period (2012-2013). This study capitalizes on the ozone profile measurements from SHADOZ to evaluate OMPS limb profile retrievals. Finally, we use SHADOZ sondes and OMPS retrievals to examine the agreement with the GEOS-5 Ozone Assimilation System (GOAS). The GOAS uses data from the OMI and the Microwave Limb Sounder (MLS) to constrain the total column and stratospheric profiles of ozone. The most recent version of the assimilation system is well constrained to the total column compared with SHADOZ ozonesonde data.

Influence of dissolved organic matter and activated carbon pore characteristics on organic micropollutant desorption.

PubMed

Aschermann, Geert; Zietzschmann, Frederik; Jekel, Martin

2018-04-15

By simulating decreasing inflow concentrations, the extent of desorption of organic micropollutants (OMP) from three activated carbons (AC) was examined in laboratory batch tests. The tested AC showed strong differences in pore size distribution and could therefore be characterized as typical micro-, meso- and macroporous AC, respectively. Adsorption and desorption conditions were varied by using drinking water (containing dissolved organic matter (DOM)) and DOM-free pure water as background solutions to examine the influence of DOM on OMP desorption for the different AC. Under ideal conditions (adsorption and desorption in pure water) adsorption of the tested OMP was found to be highly up to completely reversible for all tested AC. Under real conditions (adsorption and desorption in drinking water) additional DOM adsorption affects desorption in different ways depending on the AC pore structure. For the micro- and mesoporous AC, an increased irreversibility of OMP adsorption was found, which shows that DOM adsorption prevents OMP desorption. This could be referred to pore blockage effects that occur during the parallel adsorption of DOM and OMP. For the macroporous AC, DOM adsorption led to an enhanced OMP desorption which could be attributed to displacement processes. These results show that smaller pores tend to be blocked by DOM which hinders OMP from desorption. The overall larger pores of the macroporous AC do not get blocked which could allow (i) OMP to desorb and (ii) DOM to enter and displace OMP. Copyright © 2018 Elsevier Ltd. All rights reserved.

Immunochemical and biological characterization of outer membrane proteins of Porphyromonas endodontalis.

PubMed Central

Ogawa, T; Kuribayashi, S; Shimauchi, H; Toda, T; Hamada, S

1992-01-01

Outer membrane proteins (OMP) of Porphyromonas endodontalis HG 370 (ATCC 35406) were prepared from the cell envelope fraction of the organisms. The cell envelope that had been obtained by sonication of the whole cells was extracted in 2% lithium dodecyl sulfate and then successively chromatographed with Sephacryl S-200 HR and DEAE-Sepharose Fast Flow. Two OMP fractions, OMP-I and OMP-II, were obtained, and their immunochemical properties and induction of specific antibodies were examined. The OMP-I preparation consisted of a major protein with an apparent molecular mass of 31 kDa and other moderate to minor proteins of 40.3, 51.4, 67, and 71.6 kDa, while the OMP-II preparation contained 14-, 15.5-, 27-, and 44-kDa proteins as revealed by sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis. OMP-I was found to form hydrophilic diffusion pores by incorporation into artificial liposomes composed of egg yolk phosphatidylcholine and dicetylphosphate, indicating that OMP-I exhibited significant porin activity. However, the liposomes containing heat-denatured OMP-I were scarcely active. Spontaneous and antigen-specific immunoglobulin M (IgM)-, IgG-, and IgA-secreting spot-forming cells (SFC) enzymatically dissociated into single-cell suspensions from chronically inflamed periapical tissues and were enumerated by enzyme-linked immunospot assay. In patients with radicular cysts or dental granulomas, the major isotype of spontaneous SFC was IgG. In radicular cysts, the OMP-II-specific IgG SFC represented 0.13% of the total IgG SFC, while the antigen-specific IgA or IgM SFC was not observed. It was also found that none of these mononuclear cells produced antibodies specific for OMP-I or lipopolysaccharide of P. endodontalis. Images PMID:1328059

Porin Deficiency in Carbapenem-Resistant Enterobacter aerogenes Strains.

PubMed

Hao, Min; Ye, Meiping; Shen, Zhen; Hu, Fupin; Yang, Yang; Wu, Shi; Xu, Xiaogang; Zhu, Sihui; Qin, Xiaohua; Wang, Minggui

2018-03-13

The more frequent reports of carbapenem-resistant Enterobacteriaceae have raised the alarm for public health. Apart from the production of carbapenemases, deficiency (decreased or loss of expression) of outer membrane proteins (OMPs) has been proposed as a potentially important mechanism of carbapenem resistance. The aim of the present study was to evaluate the contribution of the major OMPs to carbapenem resistance in Enterobacter aerogenes (CREA) isolates and also investigate the role of small RNAs (sRNAs) in inducing porin-associated permeability defects. The differential expression of OMPs was analyzed in four clinical CREA isolates. omp35 and omp36 genes were further investigated by whole-genome sequencing, induction of meropenem resistance, sRNA overexpression, OMP complementation assays, and reverse transcription-quantitative PCR. All four isolates examined were deficient in omp35 and omp36. Functional restoration of these two genes confirmed their contribution to carbapenem resistance. The meropenem induction assay further revealed that porin deficiency plays a role in carbapenem resistance under antibiotic selection pressure. Single-point mutations in omp36 leading to premature stop codons were detected in two of the isolates. Elevated expression levels of the sRNAs micF and micC were detected in the other two porin-deficient isolates, which were predicted to be potential porin regulators from whole-genome sequencing. Overexpression of micF and micC downregulated the expression of Omp35 and Omp36, respectively. Porin deficiency plays an important role in carbapenem resistance among clinical E. aerogenes isolates under regulation of the sRNAs micC and micF. Furthermore, overexpression of micC and micF had a minor to no impact on carbapenem minimum inhibitory concentrations, and thus, the regulatory mechanism is likely to be complex.

Analytic energy gradients for the orbital-optimized third-order Møller-Plesset perturbation theory

NASA Astrophysics Data System (ADS)

Bozkaya, Uǧur

2013-09-01

Analytic energy gradients for the orbital-optimized third-order Møller-Plesset perturbation theory (OMP3) [U. Bozkaya, J. Chem. Phys. 135, 224103 (2011)], 10.1063/1.3665134 are presented. The OMP3 method is applied to problematic chemical systems with challenging electronic structures. The performance of the OMP3 method is compared with those of canonical second-order Møller-Plesset perturbation theory (MP2), third-order Møller-Plesset perturbation theory (MP3), coupled-cluster singles and doubles (CCSD), and coupled-cluster singles and doubles with perturbative triples [CCSD(T)] for investigating equilibrium geometries, vibrational frequencies, and open-shell reaction energies. For bond lengths, the performance of OMP3 is in between those of MP3 and CCSD. For harmonic vibrational frequencies, the OMP3 method significantly eliminates the singularities arising from the abnormal response contributions observed for MP3 in case of symmetry-breaking problems, and provides noticeably improved vibrational frequencies for open-shell molecules. For open-shell reaction energies, OMP3 exhibits a better performance than MP3 and CCSD as in case of barrier heights and radical stabilization energies. As discussed in previous studies, the OMP3 method is several times faster than CCSD in energy computations. Further, in analytic gradient computations for the CCSD method one needs to solve λ-amplitude equations, however for OMP3 one does not since λ _{ab}^{ij(1)} = t_{ij}^{ab(1)} and λ _{ab}^{ij(2)} = t_{ij}^{ab(2)}. Additionally, one needs to solve orbital Z-vector equations for CCSD, but for OMP3 orbital response contributions are zero owing to the stationary property of OMP3. Overall, for analytic gradient computations the OMP3 method is several times less expensive than CCSD (roughly ˜4-6 times). Considering the balance of computational cost and accuracy we conclude that the OMP3 method emerges as a very useful tool for the study of electronically challenging chemical systems.

Analytic energy gradients for the orbital-optimized third-order Møller-Plesset perturbation theory.

PubMed

Bozkaya, Uğur

2013-09-14

Analytic energy gradients for the orbital-optimized third-order Møller-Plesset perturbation theory (OMP3) [U. Bozkaya, J. Chem. Phys. 135, 224103 (2011)] are presented. The OMP3 method is applied to problematic chemical systems with challenging electronic structures. The performance of the OMP3 method is compared with those of canonical second-order Møller-Plesset perturbation theory (MP2), third-order Møller-Plesset perturbation theory (MP3), coupled-cluster singles and doubles (CCSD), and coupled-cluster singles and doubles with perturbative triples [CCSD(T)] for investigating equilibrium geometries, vibrational frequencies, and open-shell reaction energies. For bond lengths, the performance of OMP3 is in between those of MP3 and CCSD. For harmonic vibrational frequencies, the OMP3 method significantly eliminates the singularities arising from the abnormal response contributions observed for MP3 in case of symmetry-breaking problems, and provides noticeably improved vibrational frequencies for open-shell molecules. For open-shell reaction energies, OMP3 exhibits a better performance than MP3 and CCSD as in case of barrier heights and radical stabilization energies. As discussed in previous studies, the OMP3 method is several times faster than CCSD in energy computations. Further, in analytic gradient computations for the CCSD method one needs to solve λ-amplitude equations, however for OMP3 one does not since λ(ab)(ij(1))=t(ij)(ab(1)) and λ(ab)(ij(2))=t(ij)(ab(2)). Additionally, one needs to solve orbital Z-vector equations for CCSD, but for OMP3 orbital response contributions are zero owing to the stationary property of OMP3. Overall, for analytic gradient computations the OMP3 method is several times less expensive than CCSD (roughly ~4-6 times). Considering the balance of computational cost and accuracy we conclude that the OMP3 method emerges as a very useful tool for the study of electronically challenging chemical systems.

[Prokaryotic expression of trigeminy artificial fusion gene of Leptospira interrogans and the immunogenicity of its products].

PubMed

Luo, Dong-jiao; Qiu, Xiao-feng; Wang, Jiang; Yan, Jin; Wang, Hai-bin; Zhou, Jin-cheng; Yan, Jie

2008-11-01

To construct lipL32/1-lipL21-OmpL1/2 fusion gene of Leptospira interrogans and its prokaryotic expression system, and to identify the immunogenicity of its products. PCR using linking primers was applied to construct lipL32/1-lipL21-OmpL1/2 fusion gene and a prokaryotic expression system of the fusion gene was then established using routine genetic engineering technique. SDS-PAGE was used to examine output of the target recombinant protein rLipL32/1-LipL21-OmpL1/2. Double immunodiffusion and Western Blot assay were applied to identify immunogenicity of rLipL32/1-LipL21-OmpL1/2. lipL32/1-lipL21-OmpL1/2 fusion gene with correct sequence and its prokaryotic expression system E.coli BL21DE3pET42a-lipL32/1-lipL21-ompL1/2 was obtained in this study. The output of rLipL32/1-LipL21- OmpL1/2 after optimisation was 37.78 mg/L. The immunodiffusion titer of rabbit antiserum against rLipL32/1-LipL21-OmpL1/2 was 1:4. The rLipL32/1-LipL21-OmpL1/2 antiserum was able to recognize rLipL32/1-LipL21-OmpL1/2, rLipL32/1, rLipL21 and rOmpL1/2. Positive Western hybridization signals were found among rLipL32/1-LipL21-OmpL1/2 and rabbit antiserum against whole cell of strain 56601 and serum from patients infected with L.interrogans serogroups Icterohaemorrhagiae, Grippotyphosa, Autumnalis and Pomona. The fusion gene lipL32/1-lipL21-OmpL1/2 and its prokaryotic expression system were successfully constructed in this study. The expressed fusion protein can be used as the antigen for developing universal genetic engineering vaccine and universal serological tests of leptospirosis.

Orbital-optimized third-order Møller-Plesset perturbation theory and its spin-component and spin-opposite scaled variants: application to symmetry breaking problems.

PubMed

Bozkaya, Uğur

2011-12-14

In this research, orbital-optimized third-order Møller-Plesset perturbation theory (OMP3) and its spin-component and spin-opposite scaled variants (SCS-OMP3 and SOS-OMP3) are introduced. Using a Lagrangian-based approach, an efficient, quadratically convergent algorithm for variational optimization of the molecular orbitals (MOs) for third-order Møller-Plesset perturbation theory (MP3) is presented. Explicit equations for response density matrices, the MO gradient, and Hessian are reported in spin-orbital form. The OMP3, SCS-OMP3, and SOS-OMP3 approaches are compared with the second-order Møller-Plesset perturbation theory (MP2), MP3, coupled-cluster doubles (CCD), optimized-doubles (OD), and coupled-cluster singles and doubles (CCSD) methods. All these methods are applied to the O(4)(+), O(3), and seven diatomic molecules. Results demonstrate that the OMP3 and its variants provide significantly better vibrational frequencies than MP3, CCSD, and OD for the molecules where the symmetry-breaking problems are observed. For O(4)(+), the OMP3 prediction, 1343 cm(-1), for ω(6) (b(3u)) mode, where symmetry-breaking appears, is even better than presumably more reliable methods such as Brueckner doubles (BD), 1194 cm(-1), and OD, 1193 cm(-1), methods (the experimental value is 1320 cm(-1)). For O(3), the predictions of SCS-OMP3 (1143 cm(-1)) and SOS-OMP3 (1165 cm(-1)) are remarkably better than the more robust OD method (1282 cm(-1)); the experimental value is 1089 cm(-1). For the seven diatomics, again the SCS-OMP3 and SOS-OMP3 methods provide the lowest average errors, ∣Δω(e)∣ = 44 and ∣Δω(e)∣ = 35 cm(-1), respectively, while for OD, ∣Δω(e)∣ = 161 cm(-1)and CCSD ∣Δω(e)∣ = 106 cm(-1). Hence, the OMP3 and especially its spin-scaled variants perform much better than the MP3, CCSD, and more robust OD approaches for considered test cases. Therefore, considering both the computational cost and the reliability, SCS-OMP3 and SOS-OMP3 appear to be the best methods for the symmetry-breaking cases, based on present application results. The OMP3 method offers certain advantages: it provides reliable vibrational frequencies in case of symmetry-breaking problems, especially with spin-scaling tricks, its analytic gradients are easier to compute since there is no need to solve the coupled-perturbed equations for the orbital response, and the computation of one-electron properties are easier because there is no response contribution to the particle density matrices. The OMP3 has further advantages over standard MP3, making it promising for excited state properties via linear response theory. © 2011 American Institute of Physics

Molecular characterization of the Serratia marcescens OmpF porin, and analysis of S. marcescens OmpF and OmpC osmoregulation.

PubMed

Hutsul, J A; Worobec, E

1997-08-01

Serratia marcescens is a nosocomial pathogen with a high incidence of beta-lactam resistance. Reduced amounts of outer-membrane porins have been correlated with increased resistance to beta-lactams but only one porin, OmpC, has been characterized at the molecular level. In this study we present the molecular characterization of a second porin, OmpF, and an analysis of the expression of S. marcescens porins in response to various environmental changes. Two porins were isolated from the outer membrane using urea-SDS-PAGE and the relative amounts were shown to be influenced by the osmolarity of the medium and the presence of salicylate. From a S. marcescens genomic DNA library an 8 kb EcoRI fragment was isolated that hybridized with an oligonucleotide encoding the published N-terminal amino acid sequence of the S. marcescens 41 kDa porin. A 41 kDa protein was detected in the outer membrane of Escherichia coli NM522 carrying the cloned S. marcescens DNA. The cloned gene was sequenced and shown to code for a protein that shared 60-70% identity with other known OmpF and OmpC sequences. The upstream DNA sequence of the S. marcescens gene was similar to the corresponding E. coli ompF sequence; however, a regulatory element important in repression of E. coli ompF at high osmolarity was absent. The cloned S. marcescens OmpF in E. coli increased in expression in conditions of high osmolarity. The potential involvement of micF in the observed osmoregulation of S. marcescens porins is discussed.

Nonradiative decay dynamics of methyl-4-hydroxycinnamate and its hydrated complex revealed by picosecond pump-probe spectroscopy.

PubMed

Shimada, Daiki; Kusaka, Ryoji; Inokuchi, Yoshiya; Ehara, Masahiro; Ebata, Takayuki

2012-07-07

The lifetimes of methyl 4-hydroxycinnamate (OMpCA) and its mono-hydrated complex (OMpCA-H(2)O) in the S(1) state have been measured by picosecond pump-probe spectroscopy in a supersonic beam. For OMpCA, the lifetime of the S(1)-S(0) origin is 8-9 ps. On the other hand, the lifetime of the OMpCA-H(2)O complex at the origin is 930 ps, which is ∼100 times longer than that of OMpCA. Furthermore, in the complex the S(1) lifetime shows rapid decrease at an energy of ∼200 cm(-1) above the origin and finally becomes as short as 9 ps at ∼500 cm(-1). Theoretical calculations with a symmetry-adapted cluster-configuration interaction (SAC-CI) method suggest that the observed lifetime behavior of the two species is described by nonradiative decay dynamics involving trans → cis isomerization. That is both OMpCA and OMpCA-H(2)O in the S(1) state decay due to the trans → cis isomerization, and the large difference of the lifetimes between them is due to the difference of the isomerization potential energy curve. In OMpCA, the trans → cis isomerization occurs smoothly without a barrier on the S(1) surface, while in the OMpCA-H(2)O complex, there exists a barrier along the isomerization coordinate. The calculated barrier height of OMpCA-H(2)O is in good agreement with that observed experimentally.

The Outer Membrane Protein A (OmpA) of Y. pestis promotes intracellular survival and virulence in mice

PubMed Central

Bartra, Sara Schesser; Gong, Xin; Lorica, Cherish D.; Jain, Chaitanya; Nair, Manoj K. M.; Schifferli, Dieter; Qian, Lianfen; Li, Zhongwei; Plano, Gregory V.; Schesser, Kurt

2011-01-01

The plague bacterium Yersinia pestis has a number of well-described strategies to protect itself from both host cells and soluble factors. In an effort to identify additional anti-host factors, we employed a transposon site hybridization (TraSH)-based approach to screen 105 Y. pestis mutants in an in vitro infection system. In addition to loci encoding various components of the well-characterized type III secretion system (T3SS), our screen unambiguously identified ompA as a pro-survival gene. We go on to show that an engineered Y. pestis ΔompA strain, as well as a ΔompA strain of the closely related pathogen Y. pseudotuberculosis, have fully functioning T3SSs but are specifically defective in surviving within macrophages. Additionally, the Y. pestis ΔompA strain was outcompeted by the wild-type strain in a mouse co-infection assay. Unlike in other bacterial pathogens in which OmpA can promote adherence, invasion, or serum resistance, the OmpA of Y. pestis is restricted to enhancing intracellular survival. Our data show that OmpA of the pathogenic Yersinia is a virulence factor on par with the T3SS. PMID:22023991

The OmpL porin does not modulate redox potential in the periplasmic space of Escherichia coli.

PubMed

Sardesai, Abhijit A; Genevaux, Pierre; Schwager, Françoise; Ang, Debbie; Georgopoulos, Costa

2003-04-01

The Escherichia coli DsbA protein is the major oxidative catalyst in the periplasm. Dartigalongue et al. (EMBO J., 19, 5980-5988, 2000) reported that null mutations in the ompL gene of E.coli fully suppress all phenotypes associated with dsbA mutants, i.e. sensitivity to the reducing agent dithiothreitol (DTT) and the antibiotic benzylpenicillin, lack of motility, reduced alkaline phosphatase activity and mucoidy. They showed that OmpL is a porin and hypothesized that ompL null mutations exert their suppressive effect by preventing efflux of a putative oxidizing-reducing compound into the medium. We have repeated these experiments using two different ompL null alleles in at least three different E.coli K-12 genetic backgrounds and have failed to reproduce any of the ompL suppressive effects noted above. Also, we show that, contrary to earlier results, ompL null mutations alone do not result in partial DTT sensitivity or partial motility, nor do they appreciably affect bacterial growth rates or block propagation of the male-specific bacteriophage M13. Thus, our findings clearly demonstrate that ompL plays no perceptible role in modulating redox potential in the periplasm of E.coli.

Interaction between bacterial outer membrane proteins and periplasmic quality control factors: a kinetic partitioning mechanism.

PubMed

Wu, Si; Ge, Xi; Lv, Zhixin; Zhi, Zeyong; Chang, Zengyi; Zhao, Xin Sheng

2011-09-15

The OMPs (outer membrane proteins) of Gram-negative bacteria have to be translocated through the periplasmic space before reaching their final destination. The aqueous environment of the periplasmic space and high permeability of the outer membrane engender such a translocation process inevitably challenging. In Escherichia coli, although SurA, Skp and DegP have been identified to function in translocating OMPs across the periplasm, their precise roles and their relationship remain to be elucidated. In the present paper, by using fluorescence resonance energy transfer and single-molecule detection, we have studied the interaction between the OMP OmpC and these periplasmic quality control factors. The results of the present study reveal that the binding rate of OmpC to SurA or Skp is much faster than that to DegP, which may lead to sequential interaction between OMPs and different quality control factors. Such a kinetic partitioning mechanism for the chaperone-substrate interaction may be essential for the quality control of the biogenesis of OMPs.

Overexpression of MicA induces production of OmpC-enriched outer membrane vesicles that protect against Salmonella challenge.

PubMed

Choi, Hyun-Il; Kim, Moonjeong; Jeon, Jinseong; Han, Jin Kwan; Kim, Kwang-Sun

2017-08-26

Outer membrane vesicles (OMVs) derived from bacteria are promising candidates for subunit vaccines. Stresses that modulate the composition of outer membrane proteins (OMPs) are important for OMV synthesis. Small RNAs (sRNAs) expressed in response to stress regulate OMPs, although the mechanism underlying sRNA-mediated OMV biogenesis and its utility for developing vaccine platforms remains to be elucidated. Here, we characterized the role of a sRNA, MicA, which regulates OmpA, a major OMP involved in both production of OMVs and reactive immunity against Salmonella challenge. A Salmonella strain overexpressing MicA generated more OMVs than a control strain. In addition, OmpC was the major component of MicA-derived OMV proteins. MicA-derived OMVs induced Th1- and Th17-type immune responses in vitro and reduced Salmonella-mediated lethality in a mouse model. Thus, OmpA-regulatory sRNA-derived OMVs may facilitate production of Salmonella-protective vaccines. Copyright © 2017 Elsevier Inc. All rights reserved.

An oral vaccine based on U-Omp19 induces protection against B. abortus mucosal challenge by inducing an adaptive IL-17 immune response in mice.

PubMed

Pasquevich, Karina A; Ibañez, Andrés E; Coria, Lorena M; García Samartino, Clara; Estein, Silvia M; Zwerdling, Astrid; Barrionuevo, Paula; Oliveira, Fernanda S; Seither, Christine; Warzecha, Heribert; Oliveira, Sergio C; Giambartolomei, Guillermo H; Cassataro, Juliana

2011-01-14

As Brucella infections occur mainly through mucosal surfaces, the development of mucosal administered vaccines could be radical for the control of brucellosis. In this work we evaluated the potential of Brucella abortus 19 kDa outer membrane protein (U-Omp19) as an edible subunit vaccine against brucellosis. We investigated the protective immune response elicited against oral B. abortus infection after vaccination of mice with leaves from transgenic plants expressing U-Omp19; or with plant-made or E. coli-made purified U-Omp19. All tested U-Omp19 formulations induced protection against Brucella when orally administered without the need of adjuvants. U-Omp19 also induced protection against a systemic challenge when parenterally administered. This built-in adjuvant ability of U-Omp19 was independent of TLR4 and could be explained at least in part by its capability to activate dendritic cells in vivo. While unadjuvanted U-Omp19 intraperitoneally administered induced a specific Th1 response, following U-Omp19 oral delivery a mixed specific Th1-Th17 response was induced. Depletion of CD4(+) T cells in mice orally vaccinated with U-Omp19 resulted in a loss of the elicited protection, indicating that this cell type mediates immune protection. The role of IL-17 against Brucella infection has never been explored. In this study, we determined that if IL-17A was neutralized in vivo during the challenge period, the mucosal U-Omp19 vaccine did not confer mucosal protection. On the contrary, IL-17A neutralization during the infection did not influence at all the subsistence and growth of this bacterium in PBS-immunized mice. All together, our results indicate that an oral unadjuvanted vaccine based on U-Omp19 induces protection against a mucosal challenge with Brucella abortus by inducing an adaptive IL-17 immune response. They also indicate different and important new aspects i) IL-17 does not contribute to reduce the bacterial burden in non vaccinated mice and ii) IL-17 plays a central role in vaccine mediated anti-Brucella mucosal immunity.

Ozone Profile Retrievals from the OMPS on Suomi NPP

NASA Astrophysics Data System (ADS)

Bak, J.; Liu, X.; Kim, J. H.; Haffner, D. P.; Chance, K.; Yang, K.; Sun, K.; Gonzalez Abad, G.

2017-12-01

We verify and correct the Ozone Mapping and Profiler Suite (OMPS) Nadir Mapper (NM) L1B v2.0 data with the aim of producing accurate ozone profile retrievals using an optimal estimation based inversion method in the 302.5-340 nm fitting. The evaluation of available slit functions demonstrates that preflight-measured slit functions well represent OMPS measurements compared to derived Gaussian slit functions. Our OMPS fitting residuals contain significant wavelength and cross-track dependent biases, and thereby serious cross-track striping errors are found in preliminary retrievals, especially in the troposphere. To eliminate the systematic component of the fitting residuals, we apply "soft calibration" to OMPS radiances. With the soft calibration the amplitude of fitting residuals decreases from 1 % to 0.2 % over low/mid latitudes, and thereby the consistency of tropospheric ozone retrievals between OMPS and Ozone Monitoring Instrument (OMI) are substantially improved. A common mode correction is implemented for additional radiometric calibration, which improves retrievals especially at high latitudes where the amplitude of fitting residuals decreases by a factor of 2. We estimate the floor noise error of OMPS measurements from standard deviations of the fitting residuals. The derived error in the Huggins band ( 0.1 %) is 2 times smaller than OMI floor noise error and 2 times larger than OMPS L1B measurement error. The OMPS floor noise errors better constrain our retrievals for maximizing measurement information and stabilizing our fitting residuals. The final precision of the fitting residuals is less than 0.1 % in the low/mid latitude, with 1 degrees of freedom for signal for the tropospheric ozone, so that we meet the general requirements for successful tropospheric ozone retrievals. To assess if the quality of OMPS ozone retrievals could be acceptable for scientific use, we will characterize OMPS ozone profile retrievals, present error analysis, and validate retrievals using a reference dataset. The useful information on the vertical distribution of ozone is limited below 40 km only from OMPS NM measurements due to the absence of Hartley ozone wavelength. This shortcoming will be improved with the joint ozone profile retrieval using Nadir Profiler (NP) measurements covering the 250 to 310 nm range.

[Expression changes of major outer membrane protein antigens in Leptospira interrogans during infection and its mechanism].

PubMed

Zheng, Linli; Ge, Yumei; Hu, Weilin; Yan, Jie

2013-03-01

To determine expression changes of major outer membrane protein(OMP) antigens of Leptospira interrogans serogroup Icterohaemorrhagiae serovar Lai strain Lai during infection of human macrophages and its mechanism. OmpR encoding genes and OmpR-related histidine kinase (HK) encoding gene of L.interrogans strain Lai and their functional domains were predicted using bioinformatics technique. mRNA level changes of the leptospiral major OMP-encoding genes before and after infection of human THP-1 macrophages were detected by real-time fluorescence quantitative RT-PCR. Effects of the OmpR-encoding genes and HK-encoding gene on the expression of leptospiral OMPs during infection were determined by HK-peptide antiserum block assay and closantel inhibitive assays. The bioinformatics analysis indicated that LB015 and LB333 were referred to OmpR-encoding genes of the spirochete, while LB014 might act as a OmpR-related HK-encoding gene. After the spirochete infecting THP-1 cells, mRNA levels of leptospiral lipL21, lipL32 and lipL41 genes were rapidly and persistently down-regulated (P <0.01), whereas mRNA levels of leptospiral groEL, mce, loa22 and ligB genes were rapidly but transiently up-regulated (P<0.01). The treatment with closantel and HK-peptide antiserum partly reversed the infection-based down-regulated mRNA levels of lipL21 and lipL48 genes (P <0.01). Moreover, closantel caused a decrease of the infection-based up-regulated mRNA levels of groEL, mce, loa22 and ligB genes (P <0.01). Expression levels of L.interrogans strain Lai major OMP antigens present notable changes during infection of human macrophages. There is a group of OmpR-and HK-encoding genes which may play a major role in down-regulation of expression levels of partial OMP antigens during infection.

Recommendations from the European Working Group for Value Assessment and Funding Processes in Rare Diseases (ORPH-VAL).

PubMed

Annemans, Lieven; Aymé, Ségolène; Le Cam, Yann; Facey, Karen; Gunther, Penilla; Nicod, Elena; Reni, Michele; Roux, Jean-Louis; Schlander, Michael; Taylor, David; Tomino, Carlo; Torrent-Farnell, Josep; Upadhyaya, Sheela; Hutchings, Adam; Le Dez, Lugdivine

2017-03-10

Rare diseases are an important public health issue with high unmet need. The introduction of the EU Regulation on orphan medicinal products (OMP) has been successful in stimulating investment in the research and development of OMPs. Despite this advancement, patients do not have universal access to these new medicines. There are many factors that affect OMP uptake, but one of the most important is the difficulty of making pricing and reimbursement (P&R) decisions in rare diseases. Until now, there has been little consensus on the most appropriate assessment criteria, perspective or appraisal process. This paper proposes nine principles to help improve the consistency of OMP P&R assessment in Europe and ensure that value assessment, pricing and funding processes reflect the specificities of rare diseases and contribute to both the sustainability of healthcare systems and the sustainability of innovation in this field. These recommendations are the output of the European Working Group for Value Assessment and Funding Processes in Rare Diseases (ORPH-VAL), a collaboration between rare disease experts, patient representatives, academics, health technology assessment (HTA) practitioners, politicians and industry representatives. ORPH-VAL reached its recommendations through careful consideration of existing OMP P&R literature and through a wide consultation with expert stakeholders, including payers, regulators and patients. The principles cover four areas: OMP decision criteria, OMP decision process, OMP sustainable funding systems and European co-ordination. This paper also presents a guide to the core elements of value relevant to OMPs that should be consistently considered in all OMP appraisals. The principles outlined in this paper may be helpful in drawing together an emerging consensus on this topic and identifying areas where consistency in payer approach could be achievable and beneficial. All stakeholders have an obligation to work together to ensure that the promise of OMP's is realised.

Insights into PG-binding, conformational change, and dimerization of the OmpA C-terminal domains from Salmonella enterica serovar Typhimurium and Borrelia burgdorferi: Characterization of OmpA C-Terminal Domain

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tan, Kemin; Deatherage Kaiser, Brooke L.; Wu, Ruiying

S. Typhimurium can induce both humoral and cell-mediated responses when establishing itself in the host. These responses are primarily stimulated against the lipopolysaccharide and major outer membrane (OM) proteins of the bacterium. OmpA is one of these major OM proteins. It comprises a N-terminal eight-stranded -barrel membrane domain and a C-terminal so-called OmpA C-terminal domain (OmpACTD). The OmpACTD and its homologs are believed to bind to peptidoglycan (PG) within the periplasm, maintaining bacterial osmotic homeostasis and modulating the permeability and integrity of the outer membrane. Here we present the structures of two forms of the OmpACTD of S. Typhimurium (STOmpACTD)more » and one structure of the less-studied OmpACTD of Borrelia burgdorferi (BbOmpACTD). In the open form of STOmpACTD, an aspartic acid residue from a long 2-3 loop points into the binding pocket, suggesting that an anion group such as a carboxylate group from PG is favored at the binding site. In the closed form of STOmpACTD and in the structure of BbOmpACTD, a sulfate group from the crystallization buffer is tightly bound at the equivalent site. The differences between the closed and open forms of STOmpACTD, suggest a large conformational change that includes an extension of 3 helix by ordering a part of 2-3 loop. We suggest that the sulfate anion observed in these structures mimics the carboxylate group of PG when bound to STOmpACTD. In addition, the binding of PG or a ligand mimic may enhance dimerization of STOmpACTD, or possibly that of full length STOmpA.« less

The effect of feed water dissolved organic carbon concentration and composition on organic micropollutant removal and microbial diversity in soil columns simulating river bank filtration.

PubMed

Bertelkamp, C; van der Hoek, J P; Schoutteten, K; Hulpiau, L; Vanhaecke, L; Vanden Bussche, J; Cabo, A J; Callewaert, C; Boon, N; Löwenberg, J; Singhal, N; Verliefde, A R D

2016-02-01

This study investigated organic micropollutant (OMP) biodegradation rates in laboratory-scale soil columns simulating river bank filtration (RBF) processes. The dosed OMP mixture consisted of 11 pharmaceuticals, 6 herbicides, 2 insecticides and 1 solvent. Columns were filled with soil from a RBF site and were fed with four different organic carbon fractions (hydrophilic, hydrophobic, transphilic and river water organic matter (RWOM)). Additionally, the effect of a short-term OMP/dissolved organic carbon (DOC) shock-load (e.g. quadrupling the OMP concentrations and doubling the DOC concentration) on OMP biodegradation rates was investigated to assess the resilience of RBF systems. The results obtained in this study imply that - in contrast to what is observed for managed aquifer recharge systems operating on wastewater effluent - OMP biodegradation rates are not affected by the type of organic carbon fraction fed to the soil column, in case of stable operation. No effect of a short-term DOC shock-load on OMP biodegradation rates between the different organic carbon fractions was observed. This means that the RBF site simulated in this study is resilient towards transient higher DOC concentrations in the river water. However, a temporary OMP shock-load affected OMP biodegradation rates observed for the columns fed with the river water organic matter (RWOM) and the hydrophilic fraction of the river water organic matter. These different biodegradation rates did not correlate with any of the parameters investigated in this study (cellular adenosine triphosphate (cATP), DOC removal, specific ultraviolet absorbance (SUVA), richness/evenness of the soil microbial population or OMP category (hydrophobicity/charge). Copyright © 2015 Elsevier Ltd. All rights reserved.

HTA programme response to the challenges of dealing with orphan medicinal products: Process evaluation in selected European countries.

PubMed

Nicod, Elena; Annemans, Lieven; Bucsics, Anna; Lee, Anne; Upadhyaya, Sheela; Facey, Karen

2017-03-28

Challenges commonly encountered in HTA of orphan medicinal products (OMPs) were identified in Advance-HTA. Since then, new initiatives have been developed to specifically address issues related to HTA of OMPs. This study aimed to understand why these new HTA initiatives in England, Scotland and at European-level were established and whether they resolve the challenges of OMPs. The work of Advance-HTA was updated with a literature review and a conceptual framework of clinical, regulatory and economic challenges for OMPs was developed. The new HTA programmes were critiqued against the conceptual framework and outstanding challenges identified. The new programmes in England and Scotland recognise the challenges identified in demonstrating the value of ultra-OMPs (and OMPs) and that they require a different process to standard HTA approaches. Wider considerations of disease and treatment experiences from a multi-stakeholder standpoint are needed, combined with other measures to deal with uncertainty (e.g. managed entry agreements). While approaches to assessing this new view of value of OMPs, extending beyond cost/QALY frameworks, differ, their criteria are similar. These are complemented by a European initiative that fosters multi-stakeholder dialogue and consensus about value determinants throughout the life-cycle of an OMP. New HTA programmes specific to OMPs have been developed but questions remain about whether they sufficiently capture value and manage uncertainty in clinical practice. Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

Effects of ompA deletion on expression of type 1 fimbriae in Escherichia coli K1 strain RS218 and on the association of E. coli with human brain microvascular endothelial cells.

PubMed

Teng, Ching-Hao; Xie, Yi; Shin, Sooan; Di Cello, Francescopaolo; Paul-Satyaseela, Maneesh; Cai, Mian; Kim, Kwang Sik

2006-10-01

We have previously shown that outer membrane protein A (OmpA) and type 1 fimbriae are the bacterial determinants involved in Escherichia coli K1 binding to human brain microvascular endothelial cells (HBMEC), which constitute the blood-brain barrier. In investigating the role of OmpA in E. coli K1 binding to HBMEC, we showed for the first time that ompA deletion decreased the expression of type 1 fimbriae in E. coli K1. Decreased expression of type 1 fimbriae in the ompA deletion mutant was largely the result of driving the fim promoter toward the type 1 fimbrial phase-OFF orientation. mRNA levels of fimB and fimE were found to be decreased with the OmpA mutant compared to the parent strain. Of interest, the ompA deletion further decreased the abilities of E. coli K1 to bind to and invade HBMEC under the conditions of fixing type 1 fimbria expression in the phase-ON or phase-OFF status. These findings suggest that the decreased ability of the OmpA mutant to interact with HBMEC is not entirely due to its decreased type 1 fimbrial expression and that OmpA and type 1 fimbriae facilitate the interaction of E. coli K1 with HBMEC at least in an additive manner.

Revealing genome-scale transcriptional regulatory landscape of OmpR highlights its expanded regulatory roles under osmotic stress in Escherichia coli K-12 MG1655.

PubMed

Seo, Sang Woo; Gao, Ye; Kim, Donghyuk; Szubin, Richard; Yang, Jina; Cho, Byung-Kwan; Palsson, Bernhard O

2017-05-19

A transcription factor (TF), OmpR, plays a critical role in transcriptional regulation of the osmotic stress response in bacteria. Here, we reveal a genome-scale OmpR regulon in Escherichia coli K-12 MG1655. Integrative data analysis reveals that a total of 37 genes in 24 transcription units (TUs) belong to OmpR regulon. Among them, 26 genes show more than two-fold changes in expression level in an OmpR knock-out strain. Specifically, we find that: 1) OmpR regulates mostly membrane-located gene products involved in diverse fundamental biological processes, such as narU (encoding nitrate/nitrite transporter), ompX (encoding outer membrane protein X), and nuoN (encoding NADH:ubiquinone oxidoreductase); 2) by investigating co-regulation of entire sets of genes regulated by other stress-response TFs, stresses are surprisingly independently regulated among each other; and, 3) a detailed investigation of the physiological roles of the newly discovered OmpR regulon genes reveals that activation of narU represents a novel strategy to significantly improve osmotic stress tolerance of E. coli. Thus, the genome-scale approach to elucidating regulons comprehensively identifies regulated genes and leads to fundamental discoveries related to stress responses.

Nonradiative Decay Dynamics of METHYL-4-HYDROXYCINNAMATE and its Monohydrated Complex Revealed by Picosecond Pump-Probe Spectroscopy

NASA Astrophysics Data System (ADS)

Ebata, T.; Shimada, D.; Kusaka, R.; Inokuchi, Y.; Ehara, M.

2012-06-01

The lifetimes of methyl 4-hydroxycinnamate (OMpCA) and its mono-hydrated complex (OMpCA-H_2O) in the S_1 state have been measured by picosecond pump-probe spectroscopy in a supersonic beam. For OMpCA, the lifetime of the S_1 - S_0 origin is 8 - 9 ps. On the other hand, the lifetime of OMpCA-H_2O complex at the origin is 930 ps, which is 100 times longer than that. Furthermore, in the complex the S_1 lifetime shows rapid decrease at an energy of 200 cm-1 above the origin and becomes as short as 9 ps at 500 cm-1. Theoretical calculations with symmetry-adapted cluster-configuration interaction (SAC-CI) method suggest that in OMpCA, the trans - cis isomerization occurs smoothly without a barrier on the S_1surface, while in OMpCA-H_2O complex, there exists a barrier along the isomerization coordinate. The calculated barrier height of OMpCA-H_2O is in good agreement with that estimated from the lifetime measurements.

Serum antibodies to outer membrane proteins (OMPs) of Moraxella (Branhamella) catarrhalis in patients with bronchiectasis: identification of OMP B1 as an important antigen.

PubMed Central

Sethi, S; Hill, S L; Murphy, T F

1995-01-01

Moraxella (Branhamella) catarrhalis is a common cause of lower respiratory tract infections in adults and of otitis media in children. Little is known about the human immune response to this bacterium. In this study, immunoblot assays were performed to detect serum immunoglobulin G antibodies directed at purified outer membrane of M. catarrhalis. Twelve serum samples, two each from six patients with bronchiectasis who were persistently colonized with this organism, were tested with their homologous M. catarrhalis sputum isolates. In all the sera, the most prominent and consistent antibody response was to a minor 84-kDa outer membrane protein, OMP B1. Immunoblot adsorption assays show that these antibodies recognize surface exposed epitopes on OMP B1. Further analysis of human serum antibodies eluted from the surface of intact bacterial cells shows that these surface-exposed epitopes on OMP B1 are heterogeneous among strains of M. catarrhalis. OMP B1 is therefore an important OMP antigen on the surface of M. catarrhalis for the human immune response to infection by this bacterium. PMID:7890418

Associations of Escherichia coli K-12 OmpF trimers with rough and smooth lipopolysaccharides

DOE Office of Scientific and Technical Information (OSTI.GOV)

Diedrich, D.L.; Stein, M.A.; Schnaitman, C.A.

1990-09-01

The associations of both rough and smooth lipopolysaccharides (LPS) with the OmpF porin of Escherichia coli K-12 were examined in galE strains deleted for ompC. Transformation with pSS37 and growth with galactose conferred the ability to assemble a Shigella dysenteriae O antigen onto the core oligosaccharide of E. coli K-12 LPS. The association of LPS with OmpF trimers was assessed by staining, autoradiography of LPS specifically labeled with (1-14C)galactose, and Western immunoblotting with a monoclonal antibody specific for OmpF trimers. These techniques revealed that the migration distances and multiple banding patterns of OmpF porin trimers in sodium dodecyl sulfate-polyacrylamide gelsmore » were dictated by the chemotype of associated LPS. Expression of smooth LPS caused almost all of the trimeric OmpF to run in gels with a slower mobility than trimers from rough strains. The LPS associated with trimers from a smooth strain differed from the bulk-phase LPS by consisting almost exclusively of molecules with O antigen.« less

[Eukaryotic expression of Leptospira interrogans lipL32/1-ompL1/1 fusion gene encoding genus-specific protein antigens and the immunoreactivity of expression products].

PubMed

Yan, Jie; Zhao, Shou-feng; Mao, Ya-fei; Ruan, Ping; Luo, Yi-hui; Li, Shu-ping; Li, Li-wei

2005-01-01

To construct the eukaryotic expression system of L.interrogans lipL32/1-ompL1/1 fusion gene and to identify the immunoreactivity of expression products. PCR with linking primer was used to construct the fusion gene lipL32/1-ompL1/1. The P.pastoris eukaryotic expression system of the fusion gene, pPIC9K-lipL32/1-ompL1/1-P. pastorisGS115, was constructed after the fusion gene was cloned and sequenced. Colony with phenotype His(+)Mut(+) was isolated by using MD and MM plates and His(+) Mut(+) transformant with high resistance to G418 was screened out by using YPD plate. Using lysate of His(+) Mut(+) colony with high copies of the target gene digested with yeast lyase as the template and 5'AOX1 and 3'AOX1 as the primers, the target fusion gene in chromosome DNA of the constructed P. pastoris engineering strain was detected by PCR. Methanol in BMMY medium was used to induce the target recombinant protein rLipL32/1-rOmpL1/1 expression. rLipL32/1-rOmpL1/1 in the medium supernatant was extracted by using ammonium sulfate precipitation and Ni-NTA affinity chromatography. Output and immunoreactivity of rLipL32/1-rOmpL1/1 were measured by SDS-PAGE and Western blot methods, respectively. Amplification fragments of the obtained fusion gene lipL32/1-ompL1/1 was 1794 bp in size. The homogeneity of nucleotide and putative amino acid sequences of the fusion gene were as high as 99.94 % and 100 %, respectively, compared with the sequences of original lipL32/1 and ompL1/1 genotypes. The constructed eukaryotic expression system was able to secrete rLipL32/1-rOmpL1/1 with an output of 10 % of the total proteins in the supernatant, which located the expected position after SDS-PAGE. The rabbit anti-rLipL32/1 and anti-rOmpL1/1 sera could combine the expressed rLipL32/1-rOmpL1/1. An eukaryotic expression system with high efficiency in P.pastoris of L.interrogans lipL32/1-ompL1/1 fusion gene was successfully constructed in this study. The expressed fusion protein shows specific immunoreactivity, which can be used as a potential antigen for developing a novel vaccine of L.interrogans.

Dual Regulation of the Small RNA MicC and the Quiescent Porin OmpN in Response to Antibiotic Stress in Escherichia coli

PubMed Central

Dam, Sushovan; Pagès, Jean-Marie

2017-01-01

Antibiotic resistant Gram-negative bacteria are a serious threat for public health. The permeation of antibiotics through their outer membrane is largely dependent on porin, changes in which cause reduced drug uptake and efficacy. Escherichia coli produces two major porins, OmpF and OmpC. MicF and MicC are small non-coding RNAs (sRNAs) that modulate the expression of OmpF and OmpC, respectively. In this work, we investigated factors that lead to increased production of MicC. micC promoter region was fused to lacZ, and the reporter plasmid was transformed into E. coli MC4100 and derivative mutants. The response of micC–lacZ to antimicrobials was measured during growth over a 6 h time period. The data showed that the expression of micC was increased in the presence of β-lactam antibiotics and in an rpoE depleted mutant. Interestingly, the same conditions enhanced the activity of an ompN–lacZ fusion, suggesting a dual transcriptional regulation of micC and the quiescent adjacent ompN. Increased levels of OmpN in the presence of sub-inhibitory concentrations of chemicals could not be confirmed by Western blot analysis, except when analyzed in the absence of the sigma factor σE. We suggest that the MicC sRNA acts together with the σE envelope stress response pathway to control the OmpC/N levels in response to β-lactam antibiotics. PMID:29211019

Characterization of the protective capacity and immunogenicity of the 69-kD outer membrane protein of Bordetella pertussis

PubMed Central

1990-01-01

Immunization with the 69-kD outer membrane protein (OMP) of Bordetella pertussis protected neonatal mice against lethal respiratory challenge with B. pertussis 18323. Active immunization elicited a serum IgG anti- 69-kD OMP response at the time of challenge, with IgG anti-69-kD OMP antibodies detected in bronchoalveolar lavage fluid after challenge. Intravenous administration of BPE8, a monoclonal IgG1 anti-69-kD OMP, also protected young mice against B. pertussis challenge. Intravenously injected BPE8 was detected in the lungs of mice at the time of aerosol challenge, suggesting that the presence of specific antibody in the lungs may mediate protection. Thus the 69-kD OMP of B. pertussis is a protective antigen in mice that elicits specific serum antibody that can transude to the lung. The 69-kD OMP was detected in a preparation of a Takeda acellular vaccine by immunoblot analysis and a serum antibody response to the 69-kD OMP was observed in 18-mo-old children boosted with this preparation of Japanese acellular vaccine. Our results demonstrate that the B. pertussis 69-kD OMP is a protective antigen in animals, is immunogenic in humans, and is present in a preparation of acellular pertussis vaccine that is widely used in Japan. These findings indicate that the 69-kD OMP should be seriously considered as a candidate for inclusion in new formulations of antigenically defined acellular pertussis vaccines. PMID:2295882

The periplasmic domain of Escherichia coli outer membrane protein A can undergo a localized temperature dependent structural transition.

PubMed

Ishida, Hiroaki; Garcia-Herrero, Alicia; Vogel, Hans J

2014-12-01

Gram-negative bacteria such as Escherichia coli are surrounded by two membranes with a thin peptidoglycan (PG)-layer located in between them in the periplasmic space. The outer membrane protein A (OmpA) is a 325-residue protein and it is the major protein component of the outer membrane of E. coli. Previous structure determinations have focused on the N-terminal fragment (residues 1-171) of OmpA, which forms an eight stranded transmembrane β-barrel in the outer membrane. Consequently it was suggested that OmpA is composed of two independently folded domains in which the N-terminal β-barrel traverses the outer membrane and the C-terminal domain (residues 180-325) adopts a folded structure in the periplasmic space. However, some reports have proposed that full-length OmpA can instead refold in a temperature dependent manner into a single domain forming a larger transmembrane pore. Here, we have determined the NMR solution structure of the C-terminal periplasmic domain of E. coli OmpA (OmpA(180-325)). Our structure reveals that the C-terminal domain folds independently into a stable globular structure that is homologous to the previously reported PG-associated domain of Neisseria meningitides RmpM. Our results lend credence to the two domain structure model and a PG-binding function for OmpA, and we could indeed localize the PG-binding site on the protein through NMR chemical shift perturbation experiments. On the other hand, we found no evidence for binding of OmpA(180-325) with the TonB protein. In addition, we have also expressed and purified full-length OmpA (OmpA(1-325)) to study the structure of the full-length protein in micelles and nanodiscs by NMR spectroscopy. In both membrane mimetic environments, the recombinant OmpA maintains its two domain structure that is connected through a flexible linker. A series of temperature-dependent HSQC experiments and relaxation dispersion NMR experiments detected structural destabilization in the bulge region of the periplasmic domain of OmpA above physiological temperatures, which may induce dimerization and play a role in triggering the previously reported larger pore formation. Copyright © 2014 Elsevier B.V. All rights reserved.

Multi-barrier approach for removing organic micropollutants using mobile water treatment systems.

PubMed

Yu, Youngbeom; Choi, Yang Hun; Choi, Jaewon; Choi, Soohoon; Maeng, Sung Kyu

2018-05-20

The diversity of organic micropollutants (OMPs) in aquatic environments has been increasing rapidly during the last decade. Therefore, it is important to monitor and attenuate emerging contaminants before they can negatively affect the aquatic environment. However, due to the diversity and complexity of OMPs, there are limitations to using a single method for treating a combination of these pollutants. To address this issue, a mobile water treatment system (MWTS) equipped with different treatment units was designed to remove OMPs under field conditions. The MWTS was configured with various modular units including coagulation, flocculation, dissolved air flotation, membrane filtration, ozone oxidation, granular activated carbon, and UV disinfection. Each treatment unit could be operated either individually or in different combinations to identify the optimal configuration of treatment units for the removal of OMPs. To investigate the effectiveness of the MWTS, twelve OMPs were selected and introduced simultaneously into the feed water samples collected from different rivers throughout Korea. The current study proved that the MTWS is an effective solution to treat OMPs and is a time saving treatment system. The combined effects of the different treatment units removed over 99% of the selected OMPs, regardless of their physicochemical properties. Moreover, since the system is mobile, on-site analyses can be conducted to identify the most effective treatment method and configuration for each OMP. Copyright © 2018 Elsevier B.V. All rights reserved.

Smithsonian Astrophysical Observatory Ozone Mapping and Profiler Suite (SAO OMPS) formaldehyde retrieval

NASA Astrophysics Data System (ADS)

González Abad, Gonzalo; Vasilkov, Alexander; Seftor, Colin; Liu, Xiong; Chance, Kelly

2016-07-01

This paper presents our new formaldehyde (H2CO) retrievals, obtained from spectra recorded by the nadir instrument of the Ozone Mapping and Profiler Suite (OMPS) flown on board NASA's Suomi National Polar-orbiting Partnership (SUOMI-NPP) satellite. Our algorithm is similar to the one currently in place for the production of NASA's Ozone Monitoring Instrument (OMI) operational H2CO product. We are now able to produce a set of long-term data from two different instruments that share a similar concept and a similar retrieval approach. The ongoing overlap period between OMI and OMPS offers a perfect opportunity to study the consistency between both data sets. The different spatial and spectral resolution of the instruments is a source of discrepancy in the retrievals despite the similarity of the physic assumptions of the algorithm. We have concluded that the reduced spectral resolution of OMPS in comparison with OMI is not a significant obstacle in obtaining good-quality retrievals. Indeed, the improved signal-to-noise ratio of OMPS with respect to OMI helps to reduce the noise of the retrievals performed using OMPS spectra. However, the size of OMPS spatial pixels imposes a limitation in the capability to distinguish particular features of H2CO that are discernible with OMI. With root mean square (RMS) residuals ˜ 5 × 10-4 for individual pixels we estimate the detection limit to be about 7.5 × 1015 molecules cm-2. Total vertical column density (VCD) errors for individual pixels range between 40 % for pixels with high concentrations to 100 % or more for pixels with concentrations at or below the detection limit. We compare different OMI products (SAO OMI v3.0.2 and BIRA OMI v14) with our OMPS product using 1 year of data, between September 2012 and September 2013. The seasonality of the retrieved slant columns is captured similarly by all products but there are discrepancies in the values of the VCDs. The mean biases among the two OMI products and our OMPS product are 23 % between OMI SAO and OMPS SAO and 28 % between OMI BIRA and OMPS SAO for eight selected regions.

Physiological and Metabolic Responses Triggered by Omeprazole Improve Tomato Plant Tolerance to NaCl Stress

PubMed Central

Rouphael, Youssef; Raimondi, Giampaolo; Lucini, Luigi; Carillo, Petronia; Kyriacou, Marios C.; Colla, Giuseppe; Cirillo, Valerio; Pannico, Antonio; El-Nakhel, Christophe; De Pascale, Stefania

2018-01-01

Interest in the role of small bioactive molecules (< 500 Da) in plants is on the rise, compelled by plant scientists' attempt to unravel their mode of action implicated in stimulating growth and enhancing tolerance to environmental stressors. The current study aimed at elucidating the morphological, physiological and metabolomic changes occurring in greenhouse tomato (cv. Seny) treated with omeprazole (OMP), a benzimidazole inhibitor of animal proton pumps. The OMP was applied at three rates (0, 10, or 100 μM) as substrate drench for tomato plants grown under nonsaline (control) or saline conditions sustained by nutrient solutions of 1 or 75 mM NaCl, respectively. Increasing NaCl concentration from 1 to 75 mM decreased the tomato shoot dry weight by 49% in the 0 μM OMP treatment, whereas the reduction was not significant at 10 or 100 μM of OMP. Treatment of salinized (75 mM NaCl) tomato plants with 10 and especially 100 μM OMP decreased Na+ and Cl− while it increased Ca2+ concentration in the leaves. However, OMP was not strictly involved in ion homeostasis since the K+ to Na+ ratio did not increase under combined salinity and OMP treatment. OMP increased root dry weight, root morphological characteristics (total length and surface), transpiration, and net photosynthetic rate independently of salinity. Metabolic profiling of leaves through UHPLC liquid chromatography coupled to quadrupole-time-of-flight mass spectrometry facilitated identification of the reprogramming of a wide range of metabolites in response to OMP treatment. Hormonal changes involved an increase in ABA, decrease in auxins and cytokinin, and a tendency for GA down accumulation. Cutin biosynthesis, alteration of membrane lipids and heightened radical scavenging ability related to the accumulation of phenolics and carotenoids were observed. Several other stress-related compounds, such as polyamine conjugates, alkaloids and sesquiterpene lactones, were altered in response to OMP. Although a specific and well-defined mechanism could not be posited, the metabolic processes involved in OMP action suggest that this small bioactive molecule might have a hormone-like activity that ultimately elicits an improved tolerance to NaCl salinity stress. PMID:29535755

The OmpL37 surface-exposed protein is expressed by pathogenic Leptospira during infection and binds skin and vascular elastin.

PubMed

Pinne, Marija; Choy, Henry A; Haake, David A

2010-09-07

Pathogenic Leptospira spp. shed in the urine of reservoir hosts into freshwater can be transmitted to a susceptible host through skin abrasions or mucous membranes causing leptospirosis. The infection process involves the ability of leptospires to adhere to cell surface and extracellular matrix components, a crucial step for dissemination and colonization of host tissues. Therefore, the elucidation of novel mediators of host-pathogen interaction is important in the discovery of virulence factors involved in the pathogenesis of leptospirosis. In this study, we assess the functional roles of transmembrane outer membrane proteins OmpL36 (LIC13166), OmpL37 (LIC12263), and OmpL47 (LIC13050), which we recently identified on the leptospiral surface. We determine the capacity of these proteins to bind to host tissue components by enzyme-linked immunosorbent assay. OmpL37 binds elastin preferentially, exhibiting dose-dependent, saturating binding to human skin (K(d), 104±19 nM) and aortic elastin (K(d), 152±27 nM). It also binds fibrinogen (K(d), 244±15 nM), fibrinogen fragment D (K(d), 132±30 nM), plasma fibronectin (K(d), 359±68 nM), and murine laminin (K(d), 410±81 nM). The binding to human skin elastin by both recombinant OmpL37 and live Leptospira interrogans is specifically enhanced by rabbit antiserum for OmpL37, suggesting the involvement of OmpL37 in leptospiral binding to elastin and also the possibility that host-generated antibodies may promote rather than inhibit the adherence of leptospires to elastin-rich tissues. Further, we demonstrate that OmpL37 is recognized by acute and convalescent leptospirosis patient sera and also by Leptospira-infected hamster sera. Finally, OmpL37 protein is detected in pathogenic Leptospira serovars and not in saprophytic Leptospira. Thus, OmpL37 is a novel elastin-binding protein of pathogenic Leptospira that may be promoting attachment of Leptospira to host tissues.

Inhibition of apoptosis by Escherichia coli K1 is accompanied by increased expression of BclXL and blockade of mitochondrial cytochrome c release in macrophages.

PubMed

Sukumaran, Sunil K; Selvaraj, Suresh K; Prasadarao, Nemani V

2004-10-01

Escherichia coli K1 survival in the blood is a critical step for the onset of meningitis in neonates. Therefore, the circulating bacteria are impelled to avoid host defense mechanisms by finding a niche to survive and multiply. Our recent studies have shown that E. coli K1 enters and survives in both monocytes and macrophages in the newborn rat model of meningitis as well as in macrophage cell lines. Here we demonstrate that E. coli K1 not only extends the survival of human and murine infected macrophage cell lines but also renders them resistant to apoptosis induced by staurosporine. Macrophages infected with wild-type E. coli expressing outer membrane protein A (OmpA), but not with OmpA- E. coli, are resistant to DNA fragmentation and phosphatidylserine exposure induced by staurosporine. Infection with OmpA+ E. coli induces the expression of Bcl(XL), an antiapoptotic protein, both at the mRNA level as assessed by gene array analysis and at the protein level as evaluated by immunoblotting. OmpA- E. coli infection of macrophages induced the release of cytochrome c from mitochondria into the cytosol and the activation of caspases 3, 6, and 9, events that were significantly blocked in OmpA+ E. coli-infected macrophages. In addition, OmpA+ E. coli-infected cells were resistant to a decrease in the transmembrane potential of mitochondria induced by staurosporine as measured by the MitoCapture fluorescence technique. Complementation of OmpA- E. coli with a plasmid containing the ompA gene restored the ability of OmpA- E. coli to inhibit the apoptosis of infected macrophages, further demonstrating that E. coli OmpA expression is critical for inducing macrophage survival and thereby finding a safe haven for its growth.

Immune enhancement of Taishan Robinia pseudoacacia polysaccharide on recombinant Proteus mirabilis OmpA in chickens.

PubMed

Zhang, Yongbing; Yang, Shifa; Zhao, Xue; Yang, Ya; Li, Bing; Zhu, Fujie; Zhu, Ruiliang

2014-09-01

This study was conducted to evaluate the effects of Taishan Robinia pseudoacacia polysaccharide (TRPPS) on immune responses of chickens immunized with Proteus mirabilis outer membrane protein A (OmpA) recombinant protein vaccine. OmpA was expressed in Pichia pastoris and mixed with TRPPS. 360 chickens were randomly divided into six groups. Groups I to IV were treated with OmpA which contained TRPPS of three different dosages, Freund's adjuvant, respectively. Groups V and VI were treated with pure OmpA and physiological saline, respectively. The data showed that the antibody titers against OmpA, the concentration of IL-2, CD4 +, and CD8 +, T lymphocyte proliferation rate in Group II were significantly higher (P < 0.05) than those in the other groups, little difference in SIgA content was observed among groups I to VI. These results indicated that TRPPS strengthened humoral and cellular immune responses against recombinant OmpA vaccine. Moreover, 200 mg/mL TRPPS showed significance (P < 0.05) compared with Freund's adjuvant. Therefore, TRPPS can be developed into an adjuvant for recombinant subunit vaccine. Copyright © 2014 Elsevier B.V. All rights reserved.

Unlipidated Outer Membrane Protein Omp16 (U-Omp16) from Brucella spp. as Nasal Adjuvant Induces a Th1 Immune Response and Modulates the Th2 Allergic Response to Cow’s Milk Proteins

PubMed Central

Ibañez, Andrés E.; Smaldini, Paola; Coria, Lorena M.; Delpino, María V.; Pacífico, Lucila G. G.; Oliveira, Sergio C.; Risso, Gabriela S.; Pasquevich, Karina A.; Fossati, Carlos Alberto; Giambartolomei, Guillermo H.; Docena, Guillermo H.; Cassataro, Juliana

2013-01-01

The discovery of novel mucosal adjuvants will help to develop new formulations to control infectious and allergic diseases. In this work we demonstrate that U-Omp16 from Brucella spp. delivered by the nasal route (i.n.) induced an inflammatory immune response in bronchoalveolar lavage (BAL) and lung tissues. Nasal co-administration of U-Omp16 with the model antigen (Ag) ovalbumin (OVA) increased the amount of Ag in lung tissues and induced OVA-specific systemic IgG and T helper (Th) 1 immune responses. The usefulness of U-Omp16 was also assessed in a mouse model of food allergy. U-Omp16 i.n. administration during sensitization ameliorated the hypersensitivity responses of sensitized mice upon oral exposure to Cow’s Milk Protein (CMP), decreased clinical signs, reduced anti-CMP IgE serum antibodies and modulated the Th2 response in favor of Th1 immunity. Thus, U-Omp16 could be used as a broad Th1 mucosal adjuvant for different Ag formulations. PMID:23861971

Kelvin waves in the tropical stratosphere observed in OMPS-LP ozone measurements

NASA Astrophysics Data System (ADS)

Randel, W. J.; Park, M.

2017-12-01

We investigate equatorial waves in the tropical stratosphere using OMPS limb profiler (LP) ozone measurements spanning 2012-2017. The OMPS-LP data show clear evidence of eastward propagating planetary-scale Kelvin waves with periods near 15-20 days, and these feature are strongly modulated by the background winds linked to the quasi-biennial oscillation (QBO). We study coherence between OMPS-LP ozone and GPS radio occultation temperature measurements, and use these analyses to evaluate data quality and variability in the tropical stratosphere.

ompW is cooperatively upregulated by MarA and SoxS in response to menadione

PubMed Central

Collao, B.; Morales, E. H.; Gil, F.; Calderón, I. L.

2013-01-01

OmpW is a minor porin whose biological function has not been clearly defined. Evidence obtained in our laboratory indicates that in Salmonella enterica serovar Typhimurium the expression of OmpW is activated by SoxS upon exposure to paraquat and it is required for resistance. SoxS belongs to the AraC family of transcriptional regulators, like MarA and Rob. Due to their high structural similarity, the genes under their control have been grouped in the mar/sox/rob regulon, which presents a DNA-binding consensus sequence denominated the marsox box. In this work, we evaluated the role of the transcription factors MarA, SoxS and Rob of S. enterica serovar Typhimurium in regulating ompW expression in response to menadione. We determined the transcript and protein levels of OmpW in different genetic backgrounds; in the wild-type and Δrob strains ompW was upregulated in response to menadione, while in the ΔmarA and ΔsoxS strains the induction was abolished. In a double marA soxS mutant, ompW transcript levels were lowered after exposure to menadione, and only complementation in trans with both genes restored the positive regulation. Using transcriptional fusions and electrophoretic mobility shift assays with mutant versions of the promoter region we demonstrated that two of the predicted sites were functional. Additionally, we demonstrated that MarA increases the affinity of SoxS for the ompW promoter region. In conclusion, our study shows that ompW is upregulated in response to menadione in a cooperative manner by MarA and SoxS through a direct interaction with the promoter region. PMID:23393149

ompW is cooperatively upregulated by MarA and SoxS in response to menadione.

PubMed

Collao, B; Morales, E H; Gil, F; Calderón, I L; Saavedra, C P

2013-04-01

OmpW is a minor porin whose biological function has not been clearly defined. Evidence obtained in our laboratory indicates that in Salmonella enterica serovar Typhimurium the expression of OmpW is activated by SoxS upon exposure to paraquat and it is required for resistance. SoxS belongs to the AraC family of transcriptional regulators, like MarA and Rob. Due to their high structural similarity, the genes under their control have been grouped in the mar/sox/rob regulon, which presents a DNA-binding consensus sequence denominated the marsox box. In this work, we evaluated the role of the transcription factors MarA, SoxS and Rob of S. enterica serovar Typhimurium in regulating ompW expression in response to menadione. We determined the transcript and protein levels of OmpW in different genetic backgrounds; in the wild-type and Δrob strains ompW was upregulated in response to menadione, while in the ΔmarA and ΔsoxS strains the induction was abolished. In a double marA soxS mutant, ompW transcript levels were lowered after exposure to menadione, and only complementation in trans with both genes restored the positive regulation. Using transcriptional fusions and electrophoretic mobility shift assays with mutant versions of the promoter region we demonstrated that two of the predicted sites were functional. Additionally, we demonstrated that MarA increases the affinity of SoxS for the ompW promoter region. In conclusion, our study shows that ompW is upregulated in response to menadione in a cooperative manner by MarA and SoxS through a direct interaction with the promoter region.

Effects of long-term administration of omeprazole on bone mineral density and the mechanical properties of the bone.

PubMed

Yanagihara, Gabriela Rezende; de Paiva, Aline Goulart; Neto, Maurílio Pacheco; Torres, Larissa Helena; Shimano, Antônio Carlos; Louzada, Mário Jefferson Quirino; Annoni, Raquel; de Oliveira Penoni, Álvaro César

2015-01-01

Epidemiological studies have shown a relationship between long-term use of proton pump inhibitors and bone metabolism. However, this relationship has not yet become established. The aim of the present study was to analyze the mechanical properties and bone mineral density (BMD) of rats that were subjected to long-term omeprazole use. Fifty Wistar rats weighing between 200 and 240 g were divided equally into five groups: OMP300 (omeprazole intake at a dose of 300 μmoL/kg/day); OMP200 (200 μmoL/kg/day); OMP40 (40 μmoL/kg/day); OMP10 (10 μmoL/kg/day); and Cont (control group; intake of dilution vehicle). The solutions were administered for 90 consecutive days. After the rats had been sacrificed, their BMD, the mechanical properties of the dissected femurs and their serum Ca++ levels were analyzed. The BMD of the OMP300 group was lower than that of the controls (p = 0.006). There was no difference on comparing the OMP200, OMP40 and OMP10 groups with the controls. The maximum strength and rigidity of the femur did not differ in the experimental groups in comparison with the controls. The OMP300 group had a statistically lower serum Ca++ concentration than that of the controls (p = 0.049), but the other groups did not show any difference in relation to the controls. Daily intake of 300 μmoL/kg/day of omeprazole decreased the BMD of the femur, but without changes to the rigidity and strength of the femur in adult rats.

Continuation of Global NO2 and SO2 Monitoring with Suomi NPP OMPS

NASA Astrophysics Data System (ADS)

Yang, K.; Zhang, H.; Wang, J.; Ge, C.; Wang, Y.

2017-12-01

We have produced high-quality NO2 and SO2 standard products (named NMNO2 and NMSO2 respectively) from the SNPP OMPS-NM daily global observations. These OMPS standard products have been archived and publicly released at NASA Goddard Earth Sciences Data and Information Services Center (https://daac.gsfc.nasa.gov/information/news/595e9675624016d1af392c73/omps-nm-no- 2-and-so-2-l-2-data-products-released). Analyses and comparisons have demonstrated that the qualities of these OMPS standard products match or surpass those of the corresponding OMI products, enabling the continuity and extension of these two key standard Earth System Data Records (ESDRs) that begun with NASA's EOS Aura mission using the SNPP observations. In this presentation, we summarize the new techniques and algorithm advances that improve the accuracy and consistency of these ESDRs from satellite observations, and highlight the regional changes in NO2 and SO2 detected from half a decade of SNPP OMPS observations.

Virulence characteristics of extraintestinal pathogenic Escherichia coli deletion of gene encoding the outer membrane protein X.

PubMed

Meng, Xianrong; Liu, Xueling; Zhang, Liyuan; Hou, Bo; Li, Binyou; Tan, Chen; Li, Zili; Zhou, Rui; Li, Shaowen

2016-09-01

Outer membrane protein X (OmpX) and its homologues have been proposed to contribute to the virulence in various bacterial species. But, their role in virulence of extraintestinal pathogenic Escherichia coli (ExPEC) is yet to be determined. This study evaluates the role of OmpX in ExPEC virulence in vitro and in vivo using a clinical strain PPECC42 of porcine origin. The ompX deletion mutant exhibited increased swimming motility and decreased adhesion to, and invasion of pulmonary epithelial A549 cell, compared to the wild-type strain. A mild increase in LD50 and distinct decrease in bacterial load in such organs as heart, liver, spleen, lung and kidney were observed in mice infected with the ompX mutant. Complementation of the complete ompX gene in trans restored the virulence of mutant strain to the level of wild-type strain. Our results reveal that OmpX contributes to ExPEC virulence, but may be not an indispensable virulence determinant.

Outer Membrane Protein Folding and Topology from a Computational Transfer Free Energy Scale.

PubMed

Lin, Meishan; Gessmann, Dennis; Naveed, Hammad; Liang, Jie

2016-03-02

Knowledge of the transfer free energy of amino acids from aqueous solution to a lipid bilayer is essential for understanding membrane protein folding and for predicting membrane protein structure. Here we report a computational approach that can calculate the folding free energy of the transmembrane region of outer membrane β-barrel proteins (OMPs) by combining an empirical energy function with a reduced discrete state space model. We quantitatively analyzed the transfer free energies of 20 amino acid residues at the center of the lipid bilayer of OmpLA. Our results are in excellent agreement with the experimentally derived hydrophobicity scales. We further exhaustively calculated the transfer free energies of 20 amino acids at all positions in the TM region of OmpLA. We found that the asymmetry of the Gram-negative bacterial outer membrane as well as the TM residues of an OMP determine its functional fold in vivo. Our results suggest that the folding process of an OMP is driven by the lipid-facing residues in its hydrophobic core, and its NC-IN topology is determined by the differential stabilities of OMPs in the asymmetrical outer membrane. The folding free energy is further reduced by lipid A and assisted by general depth-dependent cooperativities that exist between polar and ionizable residues. Moreover, context-dependency of transfer free energies at specific positions in OmpLA predict regions important for protein function as well as structural anomalies. Our computational approach is fast, efficient and applicable to any OMP.

Modulation of flagellar expression in Escherichia coli by acetyl phosphate and the osmoregulator OmpR.

PubMed Central

Shin, S; Park, C

1995-01-01

During the search for unknown factors involved in motility, we have found that expression of the flagellar master operon flhDC is affected by mutations of the pta and ackA genes, encoding phosphotransacetylase and acetate kinase, respectively (S. Shin, J. Sheen, and C. Park, Korean J. Microbiol. 31:504-511, 1993). Here we describe results showing that this effect is modulated by externally added acetate, except when both pta and ackA are mutated, suggesting the role of acetyl phosphate, an intermediate of acetate metabolism, as a regulatory effector. Furthermore, the following evidence indicates that the phosphorylation of OmpR, a trans factor for osmoregulation, regulates flagellar expression. First, in a strain lacking ompR, the expression of flhDC is no longer responsive to a change in the level of acetyl phosphate. Second, an increase in medium osmolarity does not decrease flhDC expression in an ompR mutant. It is known that such an increase normally enhances OmpR phosphorylation. Third, OmpR protein binds to the DNA fragment containing the flhDC promoter, and its affinity is increased with phosphorylation by acetyl phosphate. DNase I footprinting revealed the regions of the flhDC promoter protected by OmpR in the presence or absence of phosphorylation. Therefore, we propose that the phosphorylated OmpR, generated by either osmolarity change or the internal level of acetyl phosphate, negatively regulates the expression of flagella. PMID:7642497

Modulation of flagellar expression in Escherichia coli by acetyl phosphate and the osmoregulator OmpR.

PubMed

Shin, S; Park, C

1995-08-01

During the search for unknown factors involved in motility, we have found that expression of the flagellar master operon flhDC is affected by mutations of the pta and ackA genes, encoding phosphotransacetylase and acetate kinase, respectively (S. Shin, J. Sheen, and C. Park, Korean J. Microbiol. 31:504-511, 1993). Here we describe results showing that this effect is modulated by externally added acetate, except when both pta and ackA are mutated, suggesting the role of acetyl phosphate, an intermediate of acetate metabolism, as a regulatory effector. Furthermore, the following evidence indicates that the phosphorylation of OmpR, a trans factor for osmoregulation, regulates flagellar expression. First, in a strain lacking ompR, the expression of flhDC is no longer responsive to a change in the level of acetyl phosphate. Second, an increase in medium osmolarity does not decrease flhDC expression in an ompR mutant. It is known that such an increase normally enhances OmpR phosphorylation. Third, OmpR protein binds to the DNA fragment containing the flhDC promoter, and its affinity is increased with phosphorylation by acetyl phosphate. DNase I footprinting revealed the regions of the flhDC promoter protected by OmpR in the presence or absence of phosphorylation. Therefore, we propose that the phosphorylated OmpR, generated by either osmolarity change or the internal level of acetyl phosphate, negatively regulates the expression of flagella.

Surface-exposed and antigenically conserved determinants of outer membrane proteins of Branhamella catarrhalis.

PubMed Central

Murphy, T F; Bartos, L C

1989-01-01

The outer membrane proteins (OMPs) of Branhamella catarrhalis were studied in an effort to identify surface-exposed determinants that are conserved among strains of the bacterium. Aliquots of polyclonal antiserum were absorbed individually by strains of B. catarrhalis. The absorbed antisera were tested in comparison with unabsorbed antiserum in an immunoblot assay against OMPs of the homologous strain. The absence of a band recognized by antibodies in the absorbed antiserum compared with the unabsorbed antiserum indicated that surface-exposed determinants of the absorbing strain cross-reacted with determinants on the homologous strain. Two antisera were absorbed individually by 20 strains of B. catarrhalis, and the absorbed sera were studied in this way in immunoblot assays. OMP E (molecular weight, ca. 56,000) expresses surface-exposed determinants that are shared among 17 of the 20 strains studied. Antibodies to OMP G (molecular weight, 28,000) were absorbed from both antisera by 14 of the 20 strains. These studies demonstrate that OMP E and OMP G express determinants that are exposed on the surface of the intact bacterium. Furthermore, these determinants are antigenically conserved among a majority of strains of B. catarrhalis. On the basis of these observations, OMPs E and G should be considered when bacterial antigens are evaluated as potential vaccine candidates. Images PMID:2476393

Protein transduction domain of transactivating transcriptional activator fused to outer membrane protein K of Vibrio parahaemolyticus to vaccinate marbled eels (Anguilla marmorata) confers protection against mortality caused by V. parahaemolyticus.

PubMed

Wang, Hang; Yang, Wei; Shen, Guoying; Zhang, Jianting; Lv, Wei; Ji, Binfeng; Meng, Chun

2015-07-01

Although immersion and oral vaccination are the most practical methods for fish farmers, their applications are very limited due to low immune stimulation effect. We used the protein transduction domain (PTD) of transactivating transcriptional factor (TAT) derived from HIV TAT protein to increase the delivery efficiency of aquatic protein vaccines. Vibrio parahaemolyticus outer membrane protein K (ompK), a reported vaccine candidate for the prevention of V. parahaemolyticus infection, was fused with TAT-PTD expressed in Escherichia coli. We found that PTD-ompK fusion protein effectively penetrated into marbled eel bodies. Analysis of ompK antibody titres demonstrated that immersion vaccination with PTD-ompK was superior to ompK alone and induced robust immune stimulation in marbled eels. Both active and passive protection analyses against immersive challenge with V. parahaemolyticus strains demonstrated that marbled eels immunized with PTD-ompK survived significantly longer than those immunized with ompK alone. Our results indicated that TAT-PTD could be served as is an efficient delivery system for aquatic immersion vaccinations against various infectious diseases commonly seen in aquatic farm industry. © 2015 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

Outer membrane protein a of Salmonella enterica serovar Typhimurium activates dendritic cells and enhances Th1 polarization

PubMed Central

2010-01-01

Background Typhoid, which is caused by Salmonella enterica serovar Typhimurium, remains a major health concern worldwide. Multidrug-resistant strains of Salmonella have emerged which exhibit increased survivability and virulence, thus leading to increased morbidity. However, little is known about the protective immune response against this microorganism. The outer membrane protein (Omp)A of bacteria plays an important role in pathogenesis. Results We purified OmpA from S. enterica serovar Typhimurium (OmpA-sal) and characterized the role of OmpA-sal in promoting adaptive and innate immune responses. OmpA-sal functionally activated bone marrow-derived dendritic cells by augmenting expression of CD80, CD86, and major histocompatibility complex classes I and II. Interestingly, OmpA-sal induced production of interferon-γ from T cells in mixed lymphocyte reactions, thus indicating Th1-polarizing capacity. The expression of surface markers and cytokine production in dendritic cells was mediated by the TLR4 signaling pathway in a TLR4 Knock-out system. Conclusions Our findings suggest that OmpA-sal modulates the adaptive immune responses to S. enterica serovar Typhimurium by activating dendritic cells and driving Th1 polarization, which are important properties to consider in the development of effective S. enterica serovar Typhimurium vaccines and immunotherapy adjuvant. PMID:20950448

A novel lumazine synthase molecule from Brucella significantly promotes the immune-stimulation effects of antigenic protein.

PubMed

Du, Z Q; Wang, J Y

2015-10-27

Brucella, an intracellular parasite that infects some livestock and humans, can damage or destroy the reproductive system of livestock. The syndrome is referred to as brucellosis and often occurs in pastoral areas; it is contagious from livestock to humans. In this study, the intact Brucella suis outer membrane protein 31 (omp31) gene was cloned, recombinantly expressed, and examined as a subunit vaccine candidate. The intact Brucella lumazine synthase (bls) gene was cloned and recombinantly expressed to study polymerization function in vitro. Non-reducing gel electrophoresis showed that rBs-BLS existed in different forms in vitro, including as a dimer and a pentamer. An enzyme-linked immunosorbent assay result showed that rOmp31 protein could induce production of an antibody in rabbits. However, the rOmp31-BLS fusion protein could elicit a much higher antibody titer in rabbits; this construct involved fusion of the Omp31 molecule with the BLS molecule. Our results indicate that Omp31 is involved in immune stimulation, while BLS has a polymerizing function based on rOmp31-BLS fusion protein immunogenicity. These data suggest that Omp31 is an ideal subunit vaccine candidate and that the BLS molecule is a favorable transport vector for antigenic proteins.

Use of OmpU porins for attachment and invasion of Crassostrea gigas immune cells by the oyster pathogen Vibrio splendidus

PubMed Central

Duperthuy, Marylise; Schmitt, Paulina; Garzón, Edwin; Caro, Audrey; Rosa, Rafael D.; Le Roux, Frédérique; Lautrédou-Audouy, Nicole; Got, Patrice; Romestand, Bernard; de Lorgeril, Julien; Kieffer-Jaquinod, Sylvie; Bachère, Evelyne; Destoumieux-Garzón, Delphine

2011-01-01

OmpU porins are increasingly recognized as key determinants of pathogenic host Vibrio interactions. Although mechanisms remain incompletely understood, various species, including the human pathogen Vibrio cholera, require OmpU for host colonization and virulence. We have shown previously that OmpU is essential for virulence in the oyster pathogen Vibrio splendidus LGP32. Here, we showed that V. splendidus LGP32 invades the oyster immune cells, the hemocytes, through subversion of host-cell actin cytoskeleton. In this process, OmpU serves as an adhesin/invasin required for β-integrin recognition and host cell invasion. Furthermore, the major protein of oyster plasma, the extracellular superoxide dismutase Cg-EcSOD, is used as an opsonin mediating the OmpU-promoted phagocytosis through its RGD sequence. Finally, the endocytosed bacteria were found to survive intracellularly, evading the host defense by preventing acidic vacuole formation and limiting reactive oxygen species production. We conclude that (i) V. splendidus is a facultative intracellular pathogen that manipulates host defense mechanisms to enter and survive in host immune cells, and (ii) that OmpU is a major determinant of host cell invasion in Vibrio species, used by V. splendidus LGP32 to attach and invade oyster hemocytes through opsonisation by the oyster plasma Cg-EcSOD. PMID:21282662

Chloroplast Omp85 proteins change orientation during evolution

PubMed Central

Sommer, Maik S.; Daum, Bertram; Gross, Lucia E.; Weis, Benjamin L. M.; Mirus, Oliver; Abram, Lars; Maier, Uwe-G.; Kühlbrandt, Werner; Schleiff, Enrico

2011-01-01

The majority of outer membrane proteins (OMPs) from Gram-negative bacteria and many of mitochondria and chloroplasts are β-barrels. Insertion and assembly of these proteins are catalyzed by the Omp85 protein family in a seemingly conserved process. All members of this family exhibit a characteristic N-terminal polypeptide-transport–associated (POTRA) and a C-terminal 16-stranded β-barrel domain. In plants, two phylogenetically distinct and essential Omp85's exist in the chloroplast outer membrane, namely Toc75-III and Toc75-V. Whereas Toc75-V, similar to the mitochondrial Sam50, is thought to possess the original bacterial function, its homolog, Toc75-III, evolved to the pore-forming unit of the TOC translocon for preprotein import. In all current models of OMP biogenesis and preprotein translocation, a topology of Omp85 with the POTRA domain in the periplasm or intermembrane space is assumed. Using self-assembly GFP-based in vivo experiments and in situ topology studies by electron cryotomography, we show that the POTRA domains of both Toc75-III and Toc75-V are exposed to the cytoplasm. This unexpected finding explains many experimental observations and requires a reevaluation of current models of OMP biogenesis and TOC complex function. PMID:21825140

A FRET-Based DNA Biosensor Tracks OmpR-Dependent Acidification of Salmonella during Macrophage Infection

PubMed Central

Chakraborty, Smarajit; Mizusaki, Hideaki; Kenney, Linda J.

2015-01-01

In bacteria, one paradigm for signal transduction is the two-component regulatory system, consisting of a sensor kinase (usually a membrane protein) and a response regulator (usually a DNA binding protein). The EnvZ/OmpR two-component system responds to osmotic stress and regulates expression of outer membrane proteins. In Salmonella, EnvZ/OmpR also controls expression of another two-component system SsrA/B, which is located on Salmonella Pathogenicity Island (SPI) 2. SPI-2 encodes a type III secretion system, which functions as a nanomachine to inject bacterial effector proteins into eukaryotic cells. During the intracellular phase of infection, Salmonella switches from assembling type III secretion system structural components to secreting effectors into the macrophage cytoplasm, enabling Salmonella to replicate in the phagocytic vacuole. Major questions remain regarding how bacteria survive the acidified vacuole and how acidification affects bacterial secretion. We previously reported that EnvZ sensed cytoplasmic signals rather than extracellular ones, as intracellular osmolytes altered the dynamics of a 17-amino-acid region flanking the phosphorylated histidine. We reasoned that the Salmonella cytoplasm might acidify in the macrophage vacuole to activate OmpR-dependent transcription of SPI-2 genes. To address these questions, we employed a DNA-based FRET biosensor (“I-switch”) to measure bacterial cytoplasmic pH and immunofluorescence to monitor effector secretion during infection. Surprisingly, we observed a rapid drop in bacterial cytoplasmic pH upon phagocytosis that was not predicted by current models. Cytoplasmic acidification was completely dependent on the OmpR response regulator, but did not require known OmpR-regulated genes such as ompC, ompF, or ssaC (SPI-2). Microarray analysis highlighted the cadC/BA operon, and additional experiments confirmed that it was repressed by OmpR. Acidification was blocked in the ompR null background in a Cad-dependent manner. Acid-dependent activation of OmpR stimulated type III secretion; blocking acidification resulted in a neutralized cytoplasm that was defective for SPI-2 secretion. Based upon these findings, we propose that Salmonella infection involves an acid-dependent secretion process in which the translocon SseB moves away from the bacterial cell surface as it associates with the vacuolar membrane, driving the secretion of SPI-2 effectors such as SseJ. New steps in the SPI-2 secretion process are proposed. PMID:25875623

Deuterium Labeling Together with Contrast Variation Small-angle Neutron Scattering Suggests How Skp Captures and Releases Unfolded Outer Membrane Proteins

PubMed Central

Zaccai, Nathan R.; Sandlin, Clifford W.; Hoopes, James T.; Curtis, Joseph E.; Fleming, Patrick J.; Fleming, Karen G.; Krueger, Susan

2016-01-01

In gram-negative bacteria, the chaperone protein Skp forms specific and stable complexes with membrane proteins while they are transported across the periplasm to the outer membrane. The jellyfish-like architecture of Skp is similar to the eukaryotic and archeal prefoldins and the mitochondrial Tim chaperones, that is α-helical ‘tentacles’ extend from a β-strand ‘body’ to create an internal cavity. Contrast variation small-angle neutron scattering (SANS) experiments on Skp alone in solution and bound in two different complexes to unfolded outer membrane proteins (uOMPs), OmpA and OmpW, demonstrate that the helical tentacles of Skp bind their substrate in a clamp-like mechanism in a conformation similar to that previously observed in the apo crystal structure of Skp. Deuteration of the uOMP component combined with contrast variation analysis allowed the shapes of Skp and uOMP as well as the location of uOMP with respect to Skp to be determined in both complexes. This represents unique information that could not be obtained without deuterium labeling of the uOMPs. The data yield the first direct structural evidence that the α-helical Skp tentacles move closer together on binding its substrate and that the structure of Skp is different when binding different uOMPs. This work presents, by example, a tutorial on performing SANS experiments using both deuterium labeling and contrast variation, including SANS theory, sample preparation, data collection, sample quality validation, data analysis and structure modeling. PMID:26791979

In silico local structure approach: a case study on outer membrane proteins.

PubMed

Martin, Juliette; de Brevern, Alexandre G; Camproux, Anne-Claude

2008-04-01

The detection of Outer Membrane Proteins (OMP) in whole genomes is an actual question, their sequence characteristics have thus been intensively studied. This class of protein displays a common beta-barrel architecture, formed by adjacent antiparallel strands. However, due to the lack of available structures, few structural studies have been made on this class of proteins. Here we propose a novel OMP local structure investigation, based on a structural alphabet approach, i.e., the decomposition of 3D structures using a library of four-residue protein fragments. The optimal decomposition of structures using hidden Markov model results in a specific structural alphabet of 20 fragments, six of them dedicated to the decomposition of beta-strands. This optimal alphabet, called SA20-OMP, is analyzed in details, in terms of local structures and transitions between fragments. It highlights a particular and strong organization of beta-strands as series of regular canonical structural fragments. The comparison with alphabets learned on globular structures indicates that the internal organization of OMP structures is more constrained than in globular structures. The analysis of OMP structures using SA20-OMP reveals some recurrent structural patterns. The preferred location of fragments in the distinct regions of the membrane is investigated. The study of pairwise specificity of fragments reveals that some contacts between structural fragments in beta-sheets are clearly favored whereas others are avoided. This contact specificity is stronger in OMP than in globular structures. Moreover, SA20-OMP also captured sequential information. This can be integrated in a scoring function for structural model ranking with very promising results. (c) 2007 Wiley-Liss, Inc.

Orphan drugs, orphan diseases. The first decade of orphan drug legislation in the EU.

PubMed

Joppi, Roberta; Bertele', Vittorio; Garattini, Silvio

2013-04-01

To assess the methodological quality of Orphan Medicinal Product (OMP) dossiers and discuss possible reasons for the small number of products licensed. Information about orphan drug designation, approval, refusal or withdrawal was obtained from the website of the European Medicines Agency and from the European Public Assessment Reports. From 2000 up to 2010, 80.9 % of the 845 candidate orphan drug designations received a positive opinion from the European Medicines Agency (EMA)'s Committee on Orphan Medicinal Products. Of the 108 OMP marketing authorizations applied for, 63 were granted. Randomised clinical trials were done for 38 OMPs and placebo was used as comparator for nearly half the licensed drugs. One third of the OMPs were tested in trials involving fewer than 100 patients and more than half in trials with 100-200 cases. The clinical trials lasted less than one year for 42.9 % of the approved OMPs. Although there may have been some small improvements over time in the methods for developing OMPs, in our opinion, the number of patients studied, the use of placebo as control, the type of outcome measure and the follow-up have often been inadequate. The present system should be changed to find better ways of fostering the development of effective and sustainable treatments for patients with orphan diseases. Public funds supporting independent clinical research on OMPs could bridge the gap between designation and approval. More stringent criteria to assess OMPs' efficacy and cost/effectiveness would improve the clinical value and the affordability of products allowed onto the market.

Deuterium Labeling Together with Contrast Variation Small-Angle Neutron Scattering Suggests How Skp Captures and Releases Unfolded Outer Membrane Proteins.

PubMed

Zaccai, Nathan R; Sandlin, Clifford W; Hoopes, James T; Curtis, Joseph E; Fleming, Patrick J; Fleming, Karen G; Krueger, Susan

2016-01-01

In Gram-negative bacteria, the chaperone protein Skp forms specific and stable complexes with membrane proteins while they are transported across the periplasm to the outer membrane. The jellyfish-like architecture of Skp is similar to the eukaryotic and archaeal prefoldins and the mitochondrial Tim chaperones, that is the α-helical "tentacles" extend from a β-strand "body" to create an internal cavity. Contrast variation small-angle neutron scattering (SANS) experiments on Skp alone in solution and bound in two different complexes to unfolded outer membrane proteins (uOMPs), OmpA and OmpW, demonstrate that the helical tentacles of Skp bind their substrate in a clamp-like mechanism in a conformation similar to that previously observed in the apo crystal structure of Skp. Deuteration of the uOMP component combined with contrast variation analysis allowed the shapes of Skp and uOMP as well as the location of uOMP with respect to Skp to be determined in both complexes. This represents unique information that could not be obtained without deuterium labeling of the uOMPs. The data yield the first direct structural evidence that the α-helical Skp tentacles move closer together on binding its substrate and that the structure of Skp is different when binding different uOMPs. This work presents, by example, a tutorial on performing SANS experiments using both deuterium labeling and contrast variation, including SANS theory, sample preparation, data collection, sample quality validation, data analysis, and structure modeling. © 2016 Elsevier Inc. All rights reserved.

Operations mission planner

NASA Technical Reports Server (NTRS)

Biefeld, Eric; Cooper, Lynne

1990-01-01

The findings are documented of the OMP research task, which investigated the applicability of artificial intelligence (AI) technology in support of automated scheduling. The goals of the effort are summarized and the technical accomplishments are highlighted. The OMP task succeeded in identifying how AI technology could be applied and demonstrated an AI-based automated scheduling approach through the OMP prototypes.

Isolation and characterization of OmpC porin mutants with altered pore properties

DOE Office of Scientific and Technical Information (OSTI.GOV)

Misra, R.; Benson, S.A.

1988-02-01

The LamB protien is normally required for the uptake of maltodextrins. Starting with a LamB/sup -/ OmpF/sup -/ strain, we have isolated mutants that will grow on maltodextrins. The mutation conferring the Dex/sup +/ phenotype in the majority of these mutants has been mapped to the ompC locus. These mutants, unlike LamB/sup -/ OmpF/sup -/ strains, grew on maltotriose and maltotetraose, but not on maltopentaose, and showed a significantly higher rate of (/sup 14/C) maltose uptake than the parent strain did. In addition, these mutants showed increased sensitivity to certain ..beta..-lactam antibiotics and sodium dodecyl sulfate, but did not exhibitmore » an increase in sensitivity to other antibiotics and detergents. The nucleotide sequence of these mutants has been determined. In all cases, residue 74 (arginine) of the mature OmpC protein was affected. The results suggest that this region of the OmpC protein is involved in the pore domain and that the alterations lead to an increased pore size.« less

A Bottom-Up Proteomic Approach to Identify Substrate Specificity of Outer-Membrane Protease OmpT.

PubMed

Wood, Sarah E; Sinsinbar, Gaurav; Gudlur, Sushanth; Nallani, Madhavan; Huang, Che-Fan; Liedberg, Bo; Mrksich, Milan

2017-12-22

Identifying peptide substrates that are efficiently cleaved by proteases gives insights into substrate recognition and specificity, guides development of inhibitors, and improves assay sensitivity. Peptide arrays and SAMDI mass spectrometry were used to identify a tetrapeptide substrate exhibiting high activity for the bacterial outer-membrane protease (OmpT). Analysis of protease activity for the preferred residues at the cleavage site (P1, P1') and nearest-neighbor positions (P2, P2') and their positional interdependence revealed FRRV as the optimal peptide with the highest OmpT activity. Substituting FRRV into a fragment of LL37, a natural substrate of OmpT, led to a greater than 400-fold improvement in OmpT catalytic efficiency, with a k cat /K m value of 6.1×10 6  L mol -1  s -1 . Wild-type and mutant OmpT displayed significant differences in their substrate specificities, demonstrating that even modest mutants may not be suitable substitutes for the native enzyme. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Immunoblot detection of class-specific humoral immune response to outer membrane proteins isolated from Salmonella typhi in humans with typhoid fever.

PubMed Central

Ortiz, V; Isibasi, A; García-Ortigoza, E; Kumate, J

1989-01-01

The studies reported here were undertaken to assess the ability of the outer membrane proteins (OMPs) of Salmonella typhi to induce a humoral immune response in humans with typhoid fever. OMPs were isolated with the nonionic detergent Triton X-100 and were found to be contaminated with approximately 4% lipopolysaccharide. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis patterns showed protein bands with molecular size ranges from 17 to 70 kilodaltons; the major groups of proteins were those that correspond to the porins and OmpA of gram-negative bacteria. Rabbit antiserum to OMPs or to S. typhi recognized OMPs after absorption with lipopolysaccharide. Sera from patients with typhoid fever contained immunoglobulin M antibodies which reacted with a protein of 28 kilodaltons and immunoglobulin G antibodies which reacted mainly with the porins, as determined by immunoblotting. These results indicate that the porins are the major immunogenic OMPs from S. typhi and that the immune response induced in the infection could be related to the protective status. Images PMID:2768450

Functional assay of Salmonella typhi OmpC using reconstituted large unilamellar vesicles: a general method for characterization of outer membrane proteins.

PubMed

Sundara Baalaji, N; Mathew, M K; Krishnaswamy, S

2006-10-01

The immunodominant trimeric beta-barrel outer membrane protein OmpC from Salmonella typhi, the causative agent of typhoid, has been functionally characterized here. The activity in the vesicle environment was studied in vitro using OmpC reconstituted into proteoliposomes. Passage of polysaccharides and polyethyleneglycols through OmpC has been examined to determine the permeability properties. The relative rate of neutral solute flux yields a radius of 1.1 nm for the S. typhi OmpC pore. This is almost double the pore size of Escherichia coli. This provides an example of large pore size present in the porins that form trimers as in the general bacterial porin family. The method used in this study provides a good membrane model for functional studies of porins.

Accessing Suomi NPP OMPS Products Through the GES DISC Online Data Services

NASA Astrophysics Data System (ADS)

Johnson, J. E.; Wei, J. C.; Garasimov, I.; Vollmer, B.

2017-12-01

The NASA Goddard Earth Sciences Data and Information Services Center (GES DISC) is the primary archive of the latest versions of atmospheric composition data from the Suomi National Polar-orbiting Partnership (NPP) Ozone Mapping Profiler Suite (OMPS) mission. OMPS consists of three spectrometers: a Nadir Mapper (300-420 nm) with 50×50 km2 resolution and 2600 km wide swath, a Nadir Profiler (250-310 nm) with 250×250 km2 footprint, and a three-slit Limb Profiler (290-1000 nm) making 3 vertical profiles spaced about 250 km apart with 1-2 km vertical resolution up to 65 km altitude. OMPS measures primarily ozone, both total column and vertical profiles, but also includes measurements of NO2 and SO2 total and tropospheric columns, aerosol extinction profiles. Also available from OMPS are the Level-1B calibrated and geolocated radiances. All data products are generated at the OMPS Science Investigator Processing System (SIPS) at NASA/GSFC. This presentation will provide an overview of the OMPS products available at the GES DISC archive, as well as demonstrate the various data services provided by the GES DISC. Traditionally users have accessed data by downloading data files using anonymous FTP. Although one may still download the full OMPS data products from the archive (using HTTPS instead), the GES DISC now also offers online data services that allow users to not have to physically download the full data files to their desktop computer. Users can access the data through a desktop client tool (such as IDL, Matlab or Panoply) using OPeNDAP. Other data services include file subsetters (spatially, temporally, and/or by variable), as well as data visualization and exploration services for users to preview or quickly analyze the data. Since TOMS and EOS Aura data products are also available from the GES DISC archive, these can be easily accessed and compared with the OMPS data.

Highlights from the 11-Year Record of Tropospheric Ozone from OMI/MLS and Continuation of that Long Record Using OMPS Measurements

NASA Technical Reports Server (NTRS)

Ziemke, J. R.; Kramarova, N. A.; Bhartia, P. K.; Degenstein, D. A.; Deland, M. T.

2016-01-01

Since October 2004 the Ozone Monitoring Instrument (OMI) and Microwave Limb Sounder (MLS) onboard the Aura satellite have provided over 11 years of continuous tropospheric ozone measurements. These OMI/MLS measurements have been used in many studies to evaluate dynamical and photochemical effects caused by ENSO, the Madden-Julian Oscillation (MJO) and shorter timescales, as well as long-term trends and the effects of deep convection on tropospheric ozone. Given that the OMI and MLS instruments have now extended well beyond their expected lifetimes, our goal is to continue their long record of tropospheric ozone using recent Ozone Mapping Profiler Suite (OMPS) measurements. The OMPS onboard the Suomi National Polar-orbiting Partnership NPP satellite was launched on October 28, 2011 and is comprised of three instruments: the nadir mapper, the nadir profiler, and the limb profiler. Our study combines total column ozone from the OMPS nadir mapper with stratospheric column ozone from the OMPS limb profiler to measure tropospheric ozone residual. The time period for the OMPS measurements is March 2012 present. For the OMPS limb profiler retrievals, the OMPS v2 algorithm from Goddard is tested against the University of Saskatchewan (USask) Algorithm. The retrieved ozone profiles from each of these algorithms are evaluated with ozone profiles from both ozonesondes and the Aura Microwave Limb Sounder (MLS). Effects on derived OMPS tropospheric ozone caused by the 2015-2016 El Nino event are highlighted. This recent El Nino produced anomalies in tropospheric ozone throughout the tropical Pacific involving increases of approximately 10 DU over Indonesia and decreases approximately 5-10 DU in the eastern Pacific. These changes in ozone due to El Nino were predominantly dynamically-induced, caused by the eastward shift in sea-surface temperature and convection from the western to the eastern Pacific.

Outer membrane protein P4 is not required for virulence in the human challenge model of Haemophilus ducreyi infection.

PubMed

Janowicz, Diane M; Zwickl, Beth W; Fortney, Kate R; Katz, Barry P; Bauer, Margaret E

2014-06-24

Bacterial lipoproteins often play important roles in pathogenesis and can stimulate protective immune responses. Such lipoproteins are viable vaccine candidates. Haemophilus ducreyi, which causes the sexually transmitted disease chancroid, expresses a number of lipoproteins during human infection. One such lipoprotein, OmpP4, is homologous to the outer membrane lipoprotein e (P4) of H. influenzae. In H. influenzae, e (P4) stimulates production of bactericidal and protective antibodies and contributes to pathogenesis by facilitating acquisition of the essential nutrients heme and nicotinamide adenine dinucleotide (NAD). Here, we tested the hypothesis that, like its homolog, H. ducreyi OmpP4 contributes to virulence and stimulates production of bactericidal antibodies. We determined that OmpP4 is broadly conserved among clinical isolates of H. ducreyi. We next constructed and characterized an isogenic ompP4 mutant, designated 35000HPompP4, in H. ducreyi strain 35000HP. To test whether OmpP4 was necessary for virulence in humans, eight healthy adults were experimentally infected. Each subject was inoculated with a fixed dose of 35000HP on one arm and three doses of 35000HPompP4 on the other arm. The overall parent and mutant pustule formation rates were 52.4% and 47.6%, respectively (P = 0.74). These results indicate that expression of OmpP4 in not necessary for H. ducreyi to initiate disease or progress to pustule formation in humans. Hyperimmune mouse serum raised against purified, recombinant OmpP4 did not promote bactericidal killing of 35000HP or phagocytosis by J774A.1 mouse macrophages in serum bactericidal and phagocytosis assays, respectively. Our data suggest that, unlike e (P4), H. ducreyi OmpP4 is not a suitable vaccine candidate. OmpP4 may be dispensable for virulence because of redundant mechanisms in H. ducreyi for heme acquisition and NAD utilization.

Inactivation of the olfactory marker protein (OMP) gene in river dolphins and other odontocete cetaceans.

PubMed

Springer, Mark S; Gatesy, John

2017-04-01

Various toothed whales (Odontoceti) are unique among mammals in lacking olfactory bulbs as adults and are thought to be anosmic (lacking the olfactory sense). At the molecular level, toothed whales have high percentages of pseudogenic olfactory receptor genes, but species that have been investigated to date retain an intact copy of the olfactory marker protein gene (OMP), which is highly expressed in olfactory receptor neurons and may regulate the temporal resolution of olfactory responses. One hypothesis for the retention of intact OMP in diverse odontocete lineages is that this gene is pleiotropic with additional functions that are unrelated to olfaction. Recent expression studies provide some support for this hypothesis. Here, we report OMP sequences for representatives of all extant cetacean families and provide the first molecular evidence for inactivation of this gene in vertebrates. Specifically, OMP exhibits independent inactivating mutations in six different odontocete lineages: four river dolphin genera (Platanista, Lipotes, Pontoporia, Inia), sperm whale (Physeter), and harbor porpoise (Phocoena). These results suggest that the only essential role of OMP that is maintained by natural selection is in olfaction, although a non-olfactory role for OMP cannot be ruled out for lineages that retain an intact copy of this gene. Available genome sequences from cetaceans and close outgroups provide evidence of inactivating mutations in two additional genes (CNGA2, CNGA4), which imply further pseudogenization events in the olfactory cascade of odontocetes. Selection analyses demonstrate that evolutionary constraints on all three genes (OMP, CNGA2, CNGA4) have been greatly reduced in Odontoceti, but retain a signature of purifying selection on the stem Cetacea branch and in Mysticeti (baleen whales). This pattern is compatible with the 'echolocation-priority' hypothesis for the evolution of OMP, which posits that negative selection was maintained in the common ancestor of Cetacea and was not relaxed significantly until the evolution of echolocation in Odontoceti. Copyright © 2017 Elsevier Inc. All rights reserved.

Shigella flexneri 3a Outer Membrane Protein C Epitope Is Recognized by Human Umbilical Cord Sera and Associated with Protective Activity

PubMed Central

Jarząb, Anna; Witkowska, Danuta; Ziomek, Edmund; Dąbrowska, Anna; Szewczuk, Zbigniew; Gamian, Andrzej

2013-01-01

Shigella flexneri 3a is one of the five major strains of the Shigella genus responsible for dysentery, especially among children, in regions of high poverty and poor sanitation. The outer membrane proteins (OMP) of this bacterium elicit immunological responses and are considered a prime target for vaccine development. When injected into mice they elicit a protective immunological response against a lethal dose of the pathogen. The OMPs from S. flexneri 3a were isolated and resolved by two-dimension-SDS-PAGE. Two 38-kDa spots were of particular interest since in our earlier studies OMPs of such molecular mass were found to interact with umbilical cord sera. These two spots were identified as OmpC by ESI-MS/MS spectrometry. By DNA sequencing, the ompC gene from S. flexneri 3a was identical to ompC from S. flexneri 2a [Gene Bank: 24113600]. A 3D model of OmpC was built and used to predict B-cell type (discontinuous) antigenic epitopes. Six epitopes bearing the highest score were selected and the corresponding peptides were synthesized. Only the peptides representing loop V of OmpC reacted strongly with the umbilical cord serum immunoglobulins. To determine which amino acids are essential for the antigenic activity of the epitope, the loop V was scanned with a series of dodecapeptides. The peptide RYDERY was identified as a minimal sequence for the loop V epitope. Truncation at either the C- or N-terminus rendered this peptide inactive. Apart from C-terminal tyrosine, substitution of each of the remaining five amino acids with glycine, led to a precipitous loss of immunological activity. This peptide may serve as a ligand in affinity chromatography of OmpC-specific antibodies and as a component of a vaccine designed to boost human immune defenses against enterobacterial infections. PMID:23940590

Suomi NPP OMPS limb profiler initial sensor performance assessment

NASA Astrophysics Data System (ADS)

Jaross, Glen; Chen, Grace; Kowitt, Mark; Warner, Jeremy; Xu, Philippe; Kelly, Thomas; Linda, Michael; Flittner, David

2012-11-01

Following the successful launch of the Ozone Mapping and Profiler Suite (OMPS) aboard the Suomi National Polar-orbiting Partnership (NPP) spacecraft, the NASA OMPS Limb team began an evaluation of sensor and data product performance in relation to the original goals for this instrument. Does the sensor design work as well as expected, and can limb scatter measurements by NPP OMPS and successor instruments form the basis for accurate long-term monitoring of ozone vertical profiles? While this paper does not address the latter question, the answer to the former is a qualified Yes given this early stage of the mission.

Melt-induced seismic anisotropy and magma assisted rifting in Ethiopia: Evidence from surface waves

NASA Astrophysics Data System (ADS)

Bastow, I. D.; Pilidou, S.; Kendall, J.-M.; Stuart, G. W.

2010-06-01

The East African rift in Ethiopia is unique worldwide because it captures the final stages of transition from continental rifting to seafloor spreading. A recent study there has shown that magma intrusion plays an important role during the final stages of continental breakup, but the mechanism by which it is incorporated into the extending plate remains ambiguous: wide-angle seismic data and complementary geophysical tools such as gravity analysis are not strongly sensitive to the geometry of subsurface melt intrusions. Studies of shear wave splitting in near-vertical SKS phases beneath the transitional Main Ethiopian Rift (MER) provide strong and consistent evidence for a rift-parallel fast anisotropic direction. However, it is difficult to discriminate between oriented melt pocket (OMP) and lattice preferred orientation (LPO) causes of anisotropy based on SKS study alone. The speeds of horizontally propagating Love (SH) and Rayleigh (SV) waves vary in similar fashions with azimuth for LPO- and OMP-induced anisotropy, but their relative change is distinctive for each mechanism. This diagnostic is exploited by studying the propagation of surface waves from a suite of azimuths across the MER. Anisotropy is roughly perpendicular to the absolute plate motion direction, thus ruling out anisotropy due to the slowly moving African Plate. Instead, three mechanisms for anisotropy act beneath the MER: periodic thin layering of seismically fast and slow material in the uppermost ˜10 km, OMP between ˜20-75 km depth, and olivine LPO in the upper mantle beneath. The results are explained best by a model in which low aspect ratio melt inclusions (dykes and veins) are being intruded into an extending plate during late stage breakup. The observations from Ethiopia join a growing body of evidence from rifts and passive margins worldwide that shows magma intrusion plays an important role in accommodating extension without marked crustal thinning.

Physicochemical and sensorial characteristics of noodle enriched with oyster mushroom (Pleorotus ostreatus) powder

NASA Astrophysics Data System (ADS)

Wahyono, A.; Novianti; Bakri, A.; Kasutjianingati

2018-01-01

Oyster Mushroom is a mushroom that can be used for food and medicine. It contains highly nutritious and functional substances such as statins and beta-glucan. A comprehensive evaluation of noodle-enriched with Oyster Mushroom powder has not been performed so far. In this study, we performed a comprehensive evaluation of noodle-enriched with Oyster Mushroom powder. The aim of this study was to assess the effects of enrichment of Oyster Mushroom Powder (OMP) on the quality of noodle. The quality of noodle was evaluated based on physical, chemical and sensorial characteristics. This study was done by substituting wheat flour with OMP at the level of 0 (control); 5; 7.5; 10; 12.5; and 15%. The results showed that OMP significantly affected (P<0.05) the chemical, physical and sensorial quality of enriched noodles. Increased OMP level concurrently increased ash, crude fiber content and water activity (Aw) of resulting noodles. The enrichment was significantly raised the redness and yellowness of noodles, but decreased whiteness index. In addition, the enrichment of OMP significantly affected the sensorial properties of enriched noodles including color, taste, aroma and texture. OMP enrichment can be done at the level of 5% without compromising color, aroma, and texture of noodle, but 12.5% for taste attribute. Thus, OMP enrichment is feasible to enhance the nutritional values of noodle.

Excavating the surface-associated and secretory proteome of Mycobacterium leprae for identifying vaccines and diagnostic markers relevant immunodominant epitopes.

PubMed

Rana, Aarti; Thakur, Shweta; Bhardwaj, Nupur; Kumar, Devender; Akhter, Yusuf

2016-12-01

For centuries, Mycobacterium leprae, etiological agent of leprosy, has been afflicting mankind regardless of extensive use of live-attenuated vaccines and antibiotics. Surface-associated and secretory proteins (SASPs) are attractive targets against bacteria. We have integrated biological knowledge with computational approaches and present a proteome-wide identification of SASPs. We also performed computational assignment of immunodominant epitopes as coordinates of prospective antigenic candidates in most important class of SASPs, the outer membrane proteins (OMPs). Exploiting the known protein sequence and structural characteristics shared by the SASPs from bacteria, 17 lipoproteins, 11 secretory and 19 novel OMPs (including 4 essential proteins) were identified in M. leprae As OMPs represent the most exposed antigens on the cell surface, their immunoinformatics analysis showed that the identified 19 OMPs harbor T-cell MHC class I epitopes and class II epitopes against HLA-DR alleles (54), while 15 OMPs present potential T-cell class II epitopes against HLA-DQ alleles (6) and 7 OMPs possess T-cell class II epitopes against HLA-DP alleles (5) of humans. Additionally, 11 M. leprae OMPs were found to have B-cell epitopes and these may be considered as prime candidates for the development of new immunotherapeutics against M. leprae. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Molecular characterization of a 40 kDa OmpC-like porin from Serratia marcescens.

PubMed

Hutsul, J A; Worobec, E

1994-02-01

An oligonucleotide that encodes the N-terminal portion of a 41 kDa porin of Serratia marcescens was used to probe S. marcescens UOC-51 genomic DNA. An 11 kb EcoRI fragment which hybridized with the oligonucleotide was subcloned into Escherichia coli, examined for expression, and sequenced. The product expressed by the cloned gene was 40 kDa. The nucleotide sequence has an ORF of 1.13 kb. When the deduced amino acid sequence was aligned and compared to other enterobacterial porins the cloned S. marcescens porin most closely resembled E. coli OmpC. Although we did not detect osmoregulation or thermoregulation of any porins in S. marcescens UOC-51, sequences analogous to the E. coli osmoregulator OmpR-binding regions are seen upstream to the cloned gene. We examined the regulation of the S. marcescens porin in E. coli and found that its expression increased in a high salt environment. A micF gene, whose transcriptional product functions to inhibit synthesis of OmpF by hybridizing with the ompF transcript, was also seen upstream of the S. marcescens ompC. An alignment with the E. coli micF gene revealed that the functional region of the S. marcescens micF gene is conserved. Based on the results obtained we have determined that S. marcescens UOC-51 produces a 40 kDa porin similar to the E. coli OmpC porin.

Identification and immunogenicity of immunodominant mimotopes of outer membrane protein U (OmpU) of Vibrio mimicus from phage display peptide library.

PubMed

Cen, Junyu; Liu, Xueqin; Li, Jinnian; Zhang, Ming; Wang, Wei

2013-01-01

Vibrio mimicus (V. mimicus) is the causative agent of ascites disease in aquatic animals. Outer membrane protein U (OmpU) is an important antigen of V. mimicus, but its protective epitopes are still unclear. A random 12-mer phage-displayed peptide library was used to screen and identify immunodominant mimotopes of the OmpU protein in V. mimicus by panning against purified OmpU-specific polyclonal antibody. Then the immunogenicity and immunoprotection in fish of these mimotopes was evaluated. Nine positive phage clones presented seven different 12- peptide sequences and more than 50% of them carried a consensus core motif of DSSK-P. These positive clones reacted with the target antibody and this interaction could be blocked, in a dose-dependent manner, by OmpU protein. Intraperitoneal injection of seven positive phage clones into fish induced a specific antibody response to OmpU protein. The fish immunized respectively with the positive phage clones C17, C24, C60 and C66 obtained 100% immunoprotective effect against experimental V. mimicus challenge. Taken together, these mimotopes presented by clone C17, C24, C60 and C66 were immunodominant mimotopes of the OmpU protein and exhibited a more appropriate candidate as epitope-based vaccine against V. mimicus infection in aquatic animals. Copyright © 2012 Elsevier Ltd. All rights reserved.

Employing Escherichia coli-derived outer membrane vesicles as an antigen delivery platform elicits protective immunity against Acinetobacter baumannii infection

NASA Astrophysics Data System (ADS)

Huang, Weiwei; Wang, Shijie; Yao, Yufeng; Xia, Ye; Yang, Xu; Li, Kui; Sun, Pengyan; Liu, Cunbao; Sun, Wenjia; Bai, Hongmei; Chu, Xiaojie; Li, Yang; Ma, Yanbing

2016-11-01

Outer membrane vesicles (OMVs) have proven to be highly immunogenic and induced an immune response against bacterial infection in human clinics and animal models. We sought to investigate whether engineered OMVs can be a feasible antigen-delivery platform for efficiently inducing specific antibody responses. In this study, Omp22 (an outer membrane protein of A. baumannii) was displayed on E. coli DH5α-derived OMVs (Omp22-OMVs) using recombinant gene technology. The morphological features of Omp22-OMVs were similar to those of wild-type OMVs (wtOMVs). Immunization with Omp22-OMVs induced high titers of Omp22-specific antibodies. In a murine sepsis model, Omp22-OMV immunization significantly protected mice from lethal challenge with a clinically isolated A. baumannii strain, which was evidenced by the increased survival rate of the mice, the reduced bacterial burdens in the lung, spleen, liver, kidney, and blood, and the suppressed serum levels of inflammatory cytokines. In vitro opsonophagocytosis assays showed that antiserum collected from Omp22-OMV-immunized mice had bactericidal activity against clinical isolates, which was partly specific antibody-dependent. These results strongly indicated that engineered OMVs could display a whole heterologous protein (~22 kDa) on the surface and effectively induce specific antibody responses, and thus OMVs have the potential to be a feasible vaccine platform.

Evaluation of ompA and pgtE genes in determining pathogenicity in Salmonella enterica serovar Enteritidis.

PubMed

Zhou, Y; Zhou, J; Wang, D; Gao, Q; Mu, X; Gao, S; Liu, X

2016-12-01

Salmonella enterica subsp. enterica serovar Enteritidis (S. Enteritidis) is a major causative agent of gastroenteritis in humans. This important food-borne pathogen also colonises the intestinal tracts of poultry and can spread systemically, especially in chickens. To identify the S. Enteritidis virulence genes involved in infection and colonisation of chickens, chromosomal deletion mutants of the ompA and pgtE genes, which encode essential components of omptins, were constructed. There were no significant differences between the wild-type and ompA and pgtE mutants in a series of in vitro assays, including an intracellular survival assay, survival in specific-pathogen-free (SPF) chicken serum, and in vitro competition assays. In contrast, in vivo competition assays revealed that ompA and pgtE mutants underwent attenuated growth in liver, cardiac blood, spleen, lung, and kidney compared to a wild-type strain (CVCC3378). When tested in SPF chickens, ompA or pgtE gene inactivation substantially reduced organ colonisation and delayed systemic infection compared with the wild-type strain. Colonisation was restored in S. Enteritidis mutants by reintroduction of the whole ompA or pgtE gene with the native promoters. The results of this study demonstrate that ompA and pgtE play an important role in the pathogenesis of S. Enteritidis and its ability to infect chickens. Copyright © 2016 Elsevier Ltd. All rights reserved.

[In vitro function of outer membrane protease T of Escherichia coli K1 pathogenic strain].

PubMed

Hui, Changye; Guo, Yan; Wu, Shuchi; Peng, Liang; Cao, Hong; Huang, Shenghe

2010-01-01

Plasminogen activation and antimicrobial peptide hydrolysis contribute to pathogens invasion and survival in vivo. To demonstrate the expression of outer membrane protease T in E. coli K1 pathogenic strain E44, its activity of plasminogen activator and protamine hydrolysis. After Benzamidine Sepharose Fast Flow and SOURCE 30Q chromatography, we got E44 outer membrane mixed fraction, and examined its activity of plasminogen activation with chromogenic substrate S-2251 method. An ompT deletion mutant of E44 was constructed by using the suicide vector pCVD442, termed as E44ompT. We examined 0.1 mg/mL cationic antimicrobial peptide protamine susceptibility of E44, ompT mutant strain E44ompT and E44ompT harboring pUCT, which was constructed by inserting complete ompT open reading frame into pUC13. We got about 37 kDa E44 membrane extract, which could activate plasminogen, and activation was membrane extract dose dependent. This confirmed the expression of outer membrane protease T in the outer membrane of E44. E44ompT was, more susceptible to 0.1 mg/mL protamine than E44, and E440mpT was partially complemented by pUCT. Outer membrane protease T is expressed in E. coli K1 pathogenic strain E44, and can activate plasminogen and hydrolyze protamine.

Prevalence and distribution of Chlamydia trachomatis genovars in Indian infertile patients: a pilot study.

PubMed

Rawre, Jyoti; Dhawan, Benu; Malhotra, Neena; Sreenivas, Vishnubhatla; Broor, Shobha; Chaudhry, Rama

2016-12-01

To determine the prevalence and distribution of Chlamydia trachomatis genovars in patients with infertility by PCR-RFLP and ompA gene sequencing. Prevalence of other etiological agents (viz., Ureaplasma spp. and Mycoplasma hominis) were also assessed. Endocervical swabs were collected from 477 women and urine was collected from 151 men attending the Infertility Clinic. The samples were screened for C. trachomatis by cryptic plasmid, ompA gene and nested ompA gene PCR. Genotyping was performed by PCR-RFLP and sequencing. Samples were screened for Ureaplasma spp. and M. hominis. The prevalence of C. trachomatis in infertile women and their male partners were 15.7% (75 of 477) and 10.0% (15 of 151) respectively. Secondary infertility was significantly associated with chlamydial infection. Genovar E was the most prevalent followed by genovar D and F. Twenty-four C. trachomatis strains were selected for ompA gene sequencing. No mixed infection was picked. Variability in ompA sequences was seen in 50.0%. Both PCR-RFLP and ompA gene sequencing showed concordant results. High prevalence of C. trachomatis in infertile couples warrants routine screening for C. trachomatis infection in all infertile couples. Genotyping of the ompA gene of C. trachomatis may be a valuable tool in understanding the natural history of C. trachomatis infection. © 2016 APMIS. Published by John Wiley & Sons Ltd.

UV254 absorbance as real-time monitoring and control parameter for micropollutant removal in advanced wastewater treatment with powdered activated carbon.

PubMed

Altmann, Johannes; Massa, Lukas; Sperlich, Alexander; Gnirss, Regina; Jekel, Martin

2016-05-01

This study investigates the applicability of UV absorbance measurements at 254 nm (UVA254) to serve as a simple and reliable surrogate parameter to monitor and control the removal of organic micropollutants (OMPs) in advanced wastewater treatment applying powdered activated carbon (PAC). Correlations between OMP removal and corresponding UVA254 reduction were determined in lab-scale adsorption batch tests and successfully applied to a pilot-scale PAC treatment stage to predict OMP removals in aggregate samples with good accuracy. Real-time UVA254 measurements were utilized to evaluate adapted PAC dosing strategies and proved to be effective for online monitoring of OMP removal. Furthermore, active PAC dosing control according to differential UVA254 measurements was implemented and tested. While precise removal predictions based on real-time measurements were not accurate for all OMPs, UVA254-controlled dynamic PAC dosing was capable of achieving stable OMP removals. UVA254 can serve as an effective surrogate parameter for OMP removal in technical PAC applications. Even though the applicability as control parameter to adjust PAC dosing to water quality changes might be limited to applications with fast response between PAC adjustment and adsorptive removal (e.g. direct filtration), UVA254 measurements can also be used to monitor the adsorption efficiency in more complex PAC applications. Copyright © 2016 Elsevier Ltd. All rights reserved.

Global survey of Klebsiella pneumoniae major porins from ertapenem non-susceptible isolates lacking carbapenemases.

PubMed

Wise, Mark G; Horvath, Elizabeth; Young, Katherine; Sahm, Daniel F; Kazmierczak, Krystyna M

2018-03-01

To understand the diversity of porin disruption in Klebsiella pneumoniae, the major outer membrane protein (OMP) porins, OmpK35 and OmpK36, were examined in a set of isolates that did not harbour traditional carbapenem-hydrolysing enzymes, but nevertheless tested non-susceptible to ertapenem. A world-wide collection of Klebsiella pneumoniae isolates that were part of the Study for Monitoring Antimicrobial Resistance Trends (SMART) surveillance project over the years 2008-2014 were characterised with regard to their β-lactamase gene carriage and potential permeability defects. Four hundred and eighty-seven isolates that did not carry carbapenemase genes, but were non-susceptible to ertapenem, were investigated by sequence analysis of the genes encoding OmpK35 and OmpK36. Isolates without obvious genetic lesions in either major porin gene were further examined by outer membrane protein SDS-PAGE. The majority of isolates, 83.0 % (404/487), exhibited clear genetic disruption in either or both of the ompK35 and ompK36 genes. Among the proportion of the collection with the highest ertapenem MIC value (>4 mg l -1 ), 60.5 % (115/190) showed mutation in both porin genes. Isolates without obvious genetic mutations were examined by SDS-PAGE, and 90.4 % (75/83) were found to lack or show altered expression of at least one of the major OMPs when compared to an ertapenem sensitive control strain. This study illustrates that porin deficiency in Klebsiella pneumoniae is a widespread phenomenon, and in combination with ESBLs and/or AmpC enzymes, likely accounts for the elevated ertapenem MICs observed in this study.

OmpL1 Is an Extracellular Matrix- and Plasminogen-Interacting Protein of Leptospira spp.

PubMed Central

Fernandes, Luis G. V.; Vieira, Monica L.; Kirchgatter, Karin; Alves, Ivy J.; de Morais, Zenaide M.; Vasconcellos, Silvio A.; Romero, Eliete C.

2012-01-01

Leptospirosis is a zoonosis with multisystem involvement caused by pathogenic strains of the genus Leptospira. OmpL1 is an outer membrane protein of Leptospira spp. that is expressed during infection. In this work, we investigated novel features of this protein. We describe that OmpL1 is a novel leptospiral extracellular matrix (ECM)-binding protein and a plasminogen (PLG) receptor. The recombinant protein was expressed in Escherichia coli BL21(DE3) Star/pLysS as inclusion bodies, refolded, and purified by metal-chelating chromatography. The protein presented a typical β-strand secondary structure, as evaluated by circular dichroism spectroscopy. The recombinant protein reacted with antibodies in serum samples from convalescent leptospirosis patients with a high specificity compared to serum samples from individuals with unrelated diseases. These data strengthen the usefulness of OmpL1 as a diagnostic marker of leptospirosis. The characterization of the immunogenicity of recombinant OmpL1 in inoculated BALB/c mice showed that the protein has the capacity to elicit humoral and cellular immune responses, as denoted by high antibody titers and the proliferation of lymphocytes. We demonstrate that OmpL1 has the ability to mediate attachment to laminin and plasma fibronectin, with KD (equilibrium dissociation constant) values of 2,099.93 ± 871.03 nM and 1,239.23 ± 506.85 nM, respectively. OmpL1 is also a PLG receptor, with a KD of 368.63 ± 121.23 nM, capable of generating enzymatically active plasmin. This is the first report that shows and characterizes OmpL1 as an ECM-interacting and a PLG-binding protein of Leptospira spp. that may play a role in bacterial pathogenesis when expressed during infection. PMID:22802342

OmpL1 is an extracellular matrix- and plasminogen-interacting protein of Leptospira spp.

PubMed

Fernandes, Luis G V; Vieira, Monica L; Kirchgatter, Karin; Alves, Ivy J; de Morais, Zenaide M; Vasconcellos, Silvio A; Romero, Eliete C; Nascimento, Ana L T O

2012-10-01

Leptospirosis is a zoonosis with multisystem involvement caused by pathogenic strains of the genus Leptospira. OmpL1 is an outer membrane protein of Leptospira spp. that is expressed during infection. In this work, we investigated novel features of this protein. We describe that OmpL1 is a novel leptospiral extracellular matrix (ECM)-binding protein and a plasminogen (PLG) receptor. The recombinant protein was expressed in Escherichia coli BL21(DE3) Star/pLysS as inclusion bodies, refolded, and purified by metal-chelating chromatography. The protein presented a typical β-strand secondary structure, as evaluated by circular dichroism spectroscopy. The recombinant protein reacted with antibodies in serum samples from convalescent leptospirosis patients with a high specificity compared to serum samples from individuals with unrelated diseases. These data strengthen the usefulness of OmpL1 as a diagnostic marker of leptospirosis. The characterization of the immunogenicity of recombinant OmpL1 in inoculated BALB/c mice showed that the protein has the capacity to elicit humoral and cellular immune responses, as denoted by high antibody titers and the proliferation of lymphocytes. We demonstrate that OmpL1 has the ability to mediate attachment to laminin and plasma fibronectin, with K(D) (equilibrium dissociation constant) values of 2,099.93 ± 871.03 nM and 1,239.23 ± 506.85 nM, respectively. OmpL1 is also a PLG receptor, with a K(D) of 368.63 ± 121.23 nM, capable of generating enzymatically active plasmin. This is the first report that shows and characterizes OmpL1 as an ECM-interacting and a PLG-binding protein of Leptospira spp. that may play a role in bacterial pathogenesis when expressed during infection.

Sorption and biodegradation of organic micropollutants during river bank filtration: a laboratory column study.

PubMed

Bertelkamp, C; Reungoat, J; Cornelissen, E R; Singhal, N; Reynisson, J; Cabo, A J; van der Hoek, J P; Verliefde, A R D

2014-04-01

This study investigated sorption and biodegradation behaviour of 14 organic micropollutants (OMP) in soil columns representative of the first metre (oxic conditions) of the river bank filtration (RBF) process. Breakthrough curves were modelled to differentiate between OMP sorption and biodegradation. The main objective of this study was to investigate if the OMP biodegradation rate could be related to the physico-chemical properties (charge, hydrophobicity and molecular weight) or functional groups of the OMPs. Although trends were observed between charge or hydrophobicity and the biodegradation rate for charged compounds, a statistically significant linear relationship for the complete OMP mixture could not be obtained using these physico-chemical properties. However, a statistically significant relationship was obtained between biological degradation rates and the OMP functional groups. The presence of ethers and carbonyl groups will increase biodegradability, while the presence of amines, ring structures, aliphatic ethers and sulphur will decrease biodegradability. This predictive model based on functional groups can be used by drinking water companies to make a first estimate whether a newly detected compound will be biodegraded during the first metre of RBF or that additional treatment is required. In addition, the influence of active and inactive biomass (biosorption), sand grains and the water matrix on OMP sorption was found to be negligible under the conditions investigated in this study. Retardation factors for most compounds were close to 1, indicating mobile behaviour of these compounds during soil passage. Adaptation of the biomass towards the dosed OMPs was not observed for a 6 month period, implying that new developed RBF sites might not be able to biodegrade compounds such as atrazine and sulfamethoxazole in the first few months of operation. Copyright © 2013 Elsevier Ltd. All rights reserved.

Progesterone for Symptomatic Perimenopause Treatment - Progesterone politics, physiology and potential for perimenopause.

PubMed

Prior, J C

2011-01-01

Perimenopause, women's normal midlife reproductive transition, is highly symptomatic for about 20% of women who are currently inaccurately counseled and inappropriately treated with oral contraceptives, menopausal hormone therapy or hysterectomy. About 80% of perimenopausal women experience vasomotor symptoms (VMS), 25% have menorrhagia, and about 10% experience mastalgia. The majority of women describe varying intensities of sleep, -coping or mood difficulties. Women are more symptomatic because common knowledge inaccurately says that estradiol (E2) levels are dropping/deficient. Evidence shows that with disturbed brain-ovary feedbacks, E2 levels average 26% higher and soar erratically - some women describe feeling pregnant! Also, ovulation and progesterone (P4) levels become insufficient or absent. The most symptomatic women have higher E2 and lower P4 levels. Because P4 and E2 complement/counterbalance each other's tissue effects, oral micronized P4 (OMP4 300 mg at -bedtime) is a physiological therapy for treatment-seeking, symptomatic perimenopausal women. Given cyclically (cycle d 14-27, or 14 on/off) in menstruating midlife women, OMP4 decreases cyclic VMS, improves sleep and premenstrual mastalgia. Menorrhagia is treated with ibuprofen 200mg/6h plus OMP4 cycle d 4-28. For insulin resistance, metformin plus cyclic or daily OMP4 decreases insulin resistance and weight gain. Non-responsive migraines need daily OMP4 plus usual therapies. VMS and insomnia in late perimenopause respond to daily OMP4. In summary, OMP4 is a physiology-based therapy that improves sleep, treats VMS, does not increase breast proliferation or cancer risk, increases bone formation and has beneficial cardiovascular effects. A controlled trial is testing OMP4 for perimenopausal VMS - more evidence-based data are needed.

A novel recombinant bivalent outer membrane protein of Vibrio vulnificus and Aeromonas hydrophila as a vaccine antigen of American eel (Anguilla rostrata).

PubMed

SongLin, Guo; PanPan, Lu; JianJun, Feng; JinPing, Zhao; Peng, Lin; LiHua, Duan

2015-04-01

The immogenicity of a novel vaccine antigen was evaluated after immunized American eels (Anguilla rostrata) with a recombinant bivalent expressed outer membrane protein (OMP) of Vibrio vulnificus and Aeromonas hydrophila. Three groups of eels were intraperitoneal (i.p) injected with phosphate-buffered saline (PBS group), formaline-killed-whole-cell (FKC) of A. hydrophila and V. vulnificus (FKC group) or the bivalent OMP (OMP group). On 14, 21, 28 and 42 days post-vaccination respectively, proliferation of the whole blood cells, titers of specific antibody and lysozyme activities of experimental eels were detected. On 28 day post-vaccination, eels from three groups were challenged by i.p injection of live A. hydrophila or V. vulnificus. The results showed that, compared with the PBS group, proliferation of whole blood cells in OMP group was significant enhanced on 28 days, and the serum titers of anti-A.hydrophila and anti-V. vulnificus antibody in eels of FKC and OMP group were significant increased on 14, 21 and 28d. Lysozyme Activities in serum, skin mucus, liver and kidney were significant changed between the three groups. Relative Percent Survival (RPS) after challenged A. hydrophila in KFC vs. PBS group and OMP vs. PBS group were 62.5% and 50% respectively, and the RPS challenged V. vulnificus in FKC and OMP vs. PBS group were 37.5% and 50% respectively. These results suggest that American eels immunized with the bivalent OMP would positively affect specific as well as non-specific immune parameters and protect against infection by the two pathogens in fresh water farming. Copyright © 2015 Elsevier Ltd. All rights reserved.

Oral delivery of Brucella spp. recombinant protein U-Omp16 abrogates the IgE-mediated milk allergy.

PubMed

Smaldini, Paola Lorena; Ibañez, Andrés Esteban; Fossati, Carlos Alberto; Cassataro, Juliana; Docena, Guillermo Horacio

2014-01-01

Food allergies are increasingly common disorders and no therapeutic strategies are yet approved. The unlipidated Omp16 (U-Omp16) is the outer membrane protein of 16 kDa from B. abortus and possesses a mucosal adjuvant property. In this study, we aimed to examine the U-Omp16 capacity to abrogate an allergen-specific Th2 immune response when it is administered as an oral adjuvant in a mouse model of food allergy.   Balb/c mice were sensitized with cholera toxin and cow's milk proteins (CMP) by gavage and simultaneously treated with U-Omp16 and CMP. Oral challenge with CMP was performed to evaluate the allergic status of mice. Symptoms, local (small bowel cytokine and transcription factor gene expression) and systemic (specific isotypes and spleen cell-secreted cytokines) parameters, and skin tests were done to evaluate the immune response. We found that the oral administration of U-Omp16 with CMP during sensitization dampened the allergic symptoms, with negativization of immediate skin test and increased skin DTH response. Serum specific IgE and IL-5 were inhibited and a Th1 response was promoted (specific IgG2a antibodies and CMP-induced IFN-γ secretion). We found at the mucosal site an inhibition of the gene expression corresponding to IL-13 and Gata-3, with an induction of IFN-γ and T-bet. These results indicated that the oral administration of U-Omp16 significantly controlled the allergic response in sensitized mice with a shift of the balance of Th1- and Th2-T cells toward Th1 predominance. These findings suggest that U-Omp16 may be useful as a Th1-directing adjuvant in an oral vaccine.

Oral delivery of Brucella spp. recombinant protein U-Omp16 abrogates the IgE-mediated milk allergy

PubMed Central

Smaldini, Paola Lorena; Ibañez, Andrés Esteban; Fossati, Carlos Alberto; Cassataro, Juliana; Docena, Guillermo Horacio

2014-01-01

Food allergies are increasingly common disorders and no therapeutic strategies are yet approved. The unlipidated Omp16 (U-Omp16) is the outer membrane protein of 16 kDa from B. abortus and possesses a mucosal adjuvant property. In this study, we aimed to examine the U-Omp16 capacity to abrogate an allergen-specific Th2 immune response when it is administered as an oral adjuvant in a mouse model of food allergy.   Balb/c mice were sensitized with cholera toxin and cow’s milk proteins (CMP) by gavage and simultaneously treated with U-Omp16 and CMP. Oral challenge with CMP was performed to evaluate the allergic status of mice. Symptoms, local (small bowel cytokine and transcription factor gene expression) and systemic (specific isotypes and spleen cell-secreted cytokines) parameters, and skin tests were done to evaluate the immune response. We found that the oral administration of U-Omp16 with CMP during sensitization dampened the allergic symptoms, with negativization of immediate skin test and increased skin DTH response. Serum specific IgE and IL-5 were inhibited and a Th1 response was promoted (specific IgG2a antibodies and CMP-induced IFN-γ secretion). We found at the mucosal site an inhibition of the gene expression corresponding to IL-13 and Gata-3, with an induction of IFN-γ and T-bet. These results indicated that the oral administration of U-Omp16 significantly controlled the allergic response in sensitized mice with a shift of the balance of Th1- and Th2-T cells toward Th1 predominance. These findings suggest that U-Omp16 may be useful as a Th1-directing adjuvant in an oral vaccine. PMID:25424811

Specific antibodies induced by nasally administered 40-kDa outer membrane protein of Porphyromonas gingivalis inhibits coaggregation activity of P. gingivalis.

PubMed

Namikoshi, Jun; Otake, Shigeo; Maeba, Satomi; Hayakawa, Mitsuo; Abiko, Yoshimitsu; Yamamoto, Masafumi

2003-12-12

In this study, we have assessed the efficacy of the 40-kDa outer membrane protein (40k-OMP) of Porphyromonas gingivalis as a nasal vaccine for the prevention of adult periodontitis. Mice nasally immunized with 40k-OMP and cholera toxin as mucosal adjuvant displayed significant levels of 40k-OMP-specific serum IgG1, IgG2b and IgA as well as mucosal IgA antibodies (Abs) in saliva and nasal secretions. Ab-forming cell (AFC) analysis confirmed the antibody titers by detecting high numbers of 40k-OMP-specific AFCs in spleen, salivary glands and nasal passages. Because 40k-OMP-specific IgG inhibited coaggregation of P. gingivalis vesicles and S. gordonii, it may be an important tool for the prevention of adult periodontitis.

Master Amino acid Pattern as substitute for dietary proteins during a weight-loss diet to achieve the body's nitrogen balance equilibrium with essentially no calories.

PubMed

Lucà-Moretti, M; Grandi, A; Lucà, E; Muratori, G; Nofroni, M G; Mucci, M P; Gambetta, P; Stimolo, R; Drago, P; Giudice, G; Tamburlin, N

2003-01-01

Results of this multicentric study have shown that by giving 10 g (10 tablets) of Master Amino acid Pattern (MAP) as a substitute for dietary proteins, once a day, to 114 overweight participants undergoing the American Nutrition Clinics/Overweight Management Program (ANC/OMP), the participants' nitrogen balance could be maintained in equilibrium with essentially no calories (MAP 1 g=0.04 kcal), thereby preserving the body's structural and functional proteins, eliminating excessive water retention from the interstitial compartment, and preventing the sudden weight increase after study conclusion commonly known as the yo-yo effect. Study results have shown that the use of MAP, in conjunction with the ANC/OMP, has proven to be safe and effective by preventing those adverse effects associated with a negative nitrogen balance, such as oversized or flabby tissue, stretch marks, sagging of breast tissue, increased hair loss, faded hair color, and fragile or brittle nails. Also preventing those anomalies commonly associated with weight-loss diets, such as hunger, weakness, headache caused by ketosis, constipation, or decreased libido, the use of MAP, in conjunction with the ANC/OMP, allowed for mean weight loss of 1.4 kg (3 lb) per week.

Human Immune Response to Outer Membrane Protein CD of Moraxella catarrhalis in Adults with Chronic Obstructive Pulmonary Disease

PubMed Central

Murphy, Timothy F.; Kirkham, Charmaine; Liu, Dai-Fang; Sethi, Sanjay

2003-01-01

Moraxella catarrhalis is a common cause of lower respiratory tract infection in adults with chronic obstructive pulmonary disease (COPD). The antibody response to outer membrane protein (OMP) CD, a highly conserved surface protein of M. catarrhalis under consideration as a vaccine antigen, was studied in adults with COPD following 40 episodes of infection or colonization. Following infection or colonization, 9 of 40 patients developed new serum immunoglobulin G (IgG) to OMP CD, as measured by enzyme-linked immunosorbent assay. Adsorption assays revealed that a proportion of the serum IgG was directed toward surface-exposed epitopes on OMP CD in six of the nine patients who developed new IgG to OMP CD. Immunoblot assays with fusion peptide constructs indicated that the new antibodies that developed after infection or colonization recognized conformational epitopes, particularly in the carboxy region of the protein. Three of 28 patients developed new mucosal IgA to OMP CD in sputum supernatants. This study establishes that OMP CD is a target of a systemic and mucosal immune response following infection and colonization in some patients with COPD. PMID:12595444

Expression of chimeric ras protein with OmpF signal peptide in Escherichia coli: localization of OmpF fusion protein in the inner membrane.

PubMed

Yamamoto, T; Okawa, N; Endo, T; Kaji, A

1991-08-01

The ras gene was fused with the DNA sequence of OmpF signal peptide or with the DNA sequence of OmpF signal peptide plus the amino terminal portion of the OmpF gene. They were placed in plasmids together with the bacteriophage lambda PL promoter. These plasmids were introduced into Escherichia coli strain K-12 and the OmpF signal peptide fusion proteins were expressed. These fusion proteins were identified as 29.0 and 30.0 kDa proteins. However, processed products of these proteins were not found in the extract. The fusion proteins were localized mostly in the cytoplasm and the inner membrane, but none of them was secreted into the periplasmic space. On the other hand, the ras protein alone was found in the cytoplasm and not in the inner membrane. Viable counts of E. coli harbouring these plasmids decreased when these fused proteins were induced. Induction of the ras protein alone did not harm cells. These observations suggest that insertion of the heterologous proteins into the inner membrane may cause the bactericidal effect.

Outer membrane proteins of pathogenic spirochetes

PubMed Central

Cullen, Paul A.; Haake, David A.; Adler, Ben

2009-01-01

Pathogenic spirochetes are the causative agents of several important diseases including syphilis, Lyme disease, leptospirosis, swine dysentery, periodontal disease and some forms of relapsing fever. Spirochetal bacteria possess two membranes and the proteins present in the outer membrane are at the site of interaction with host tissue and the immune system. This review describes the current knowledge in the field of spirochetal outer membrane protein (OMP) biology. What is known concerning biogenesis and structure of OMPs, with particular regard to the atypical signal peptide cleavage sites observed amongst the spirochetes, is discussed. We examine the functions that have been determined for several spirochetal OMPs including those that have been demonstrated to function as adhesins, porins or to have roles in complement resistance. A detailed description of the role of spirochetal OMPs in immunity, including those that stimulate protective immunity or that are involved in antigenic variation, is given. A final section is included which covers experimental considerations in spirochetal outer membrane biology. This section covers contentious issues concerning cellular localization of putative OMPs, including determination of surface exposure. A more detailed knowledge of spirochetal OMP biology will hopefully lead to the design of new vaccines and a better understanding of spirochetal pathogenesis. PMID:15449605

Genetic improvement of butanol tolerance in Escherichia coli by cell surface expression of fish metallothionein.

PubMed

Lin, Kuo Hsing; Chin, Wei Chih; Lee, Ang Hsuan; Huang, Chieh Chen

2011-01-01

Cysteine-rich metallothioneins (MTs) have been reported to possess the capacity to scavenge reactive oxygen species in vitro and in vivo. Recombinant strains of Escherichia coli expressing outer membrane protein C (OmpC) fused with MTs from human, mouse and tilapia displayed the ability for such surface-localized MTs to scavenge extracellular free radicals, but the benefits of the possible applications of this capacity have not yet been demonstrated. Because the intrinsic butanol tolerance of microbes has become an impediment for biological butanol production, we examined whether surface-displayed MTs could contribute to butanol tolerance. The results show that strains expressing OmpC-MT fusion proteins had higher butanol tolerance than strains with cytoplasmically expressed MTs. Furthermore, the OmpC-tilapia MT fusion protein enhanced butanol tolerance more strongly than other recombinant constructs. Although the enhanced level of tolerance was not as high as that provided by OmpC-tilapia MT, over-expression of OmpC was also found to contribute to butanol tolerance. These results suggest that free-radical scavenging by MT and OmpC-related osmoregulation enhance butanol tolerance. Our results shed new light on methods for engineering bacteria with higher butanol tolerance. © 2011 Landes Bioscience

Outer membrane protein A of Acinetobacter baumannii induces differentiation of CD4+ T cells toward a Th1 polarizing phenotype through the activation of dendritic cells.

PubMed

Lee, Jun Sik; Lee, Je Chul; Lee, Chang-Min; Jung, In Duk; Jeong, Young-Il; Seong, Eun-Young; Chung, Hae-Young; Park, Yeong-Min

2007-06-30

Acinetobacter baumannii is an increasing hospital-acquired pathogen that causes a various type of infections, but little is known about the protective immune response to this microorganism. Outer membrane protein A of A. baumannii (AbOmpA) is a major porin protein and plays an important role in pathogenesis. We analyzed interaction between AbOmpA and dendritic cells (DCs) to characterize the role of this protein in promoting innate and adaptive immune responses. AbOmpA functionally activates bone marrow-derived DCs by augmenting expression of the surface markers, CD40, CD54, B7 family (CD80 and CD86) and major histocompatibility complex class I and II. AbOmpA induces production of Th1-promoting interleukin-12 from DCs and augments the syngeneic and allogeneic immunostimulatory capacity of DCs. AbOmpA stimulates production of interferon-gamma from T cells in mixed lymphocyte reactions, which suggesting Th1-polarizing capacity. CD4(+) T cells stimulated by AbOmpA-stimulated DCs show a Th1-polarizing cytokine profile. The expression of surface markers on DCs is mediated by both mitogen-activated protein kinases and NF-kappaB pathways. Our findings suggest that AbOmpA induces maturation of DCs and drives Th1 polarization, which are important properties for determining the nature of immune response against A. baumannii.

Understanding the sorption and biotransformation of organic micropollutants in innovative biological wastewater treatment technologies.

PubMed

Alvarino, T; Suarez, S; Lema, J; Omil, F

2018-02-15

New technologies for wastewater treatment have been developed in the last years based on the combination of biological reactors operating under different redox conditions. Their efficiency in the removal of organic micropollutants (OMPs) has not been clearly assessed yet. This review paper is focussed on understanding the sorption and biotransformation of a selected group of 17 OMPs, including pharmaceuticals, hormones and personal care products, during biological wastewater treatment processes. Apart from considering the role of "classical" operational parameters, new factors such as biomass conformation and particle size, upward velocity applied or the addition of adsorbents have been considered. It has been found that the OMP removal by sorption not only depends on their physico-chemical characteristics and other parameters, such as the biomass conformation and particle size, or some operational conditions also relevant. Membrane biological reactors (MBR), have shown to enhance sorption and biotransformation of some OMPs. The same applies to technologies bases on direct addition of activated carbon in bioreactors. The OMP biotransformation degree and pathway is mainly driven by the redox potential and the primary substrate activity. The combination of different redox potentials in hybrid reactor systems can significantly enhance the overall OMP removal efficiency. Sorption and biotransformation can be synergistically promoted in biological reactors by the addition of activated carbon. The deeper knowledge of the main parameters influencing OMP removal provided by this review will allow optimizing the biological processes in the future. Copyright © 2017 Elsevier B.V. All rights reserved.

Trends of the Major Porin Gene (ompF) Evolution: Insight from the Genus Yersinia

PubMed Central

Stenkova, Anna M.; Isaeva, Marina P.; Shubin, Felix N.; Rasskazov, Valeri A.; Rakin, Alexander V.

2011-01-01

OmpF is one of the major general porins of Enterobacteriaceae that belongs to the first line of bacterial defense and interactions with the biotic as well as abiotic environments. Porins are surface exposed and their structures strongly reflect the history of multiple interactions with the environmental challenges. Unfortunately, little is known on diversity of porin genes of Enterobacteriaceae and the genus Yersinia especially. We analyzed the sequences of the ompF gene from 73 Yersinia strains covering 14 known species. The phylogenetic analysis placed most of the Yersinia strains in the same line assigned by 16S rDNA-gyrB tree. Very high congruence in the tree topologies was observed for Y. enterocolitica, Y. kristensenii, Y. ruckeri, indicating that intragenic recombination in these species had no effect on the ompF gene. A significant level of intra- and interspecies recombination was found for Y. aleksiciae, Y. intermedia and Y. mollaretii. Our analysis shows that the ompF gene of Yersinia has evolved with nonrandom mutational rate under purifying selection. However, several surface loops in the OmpF porin contain positively selected sites, which very likely reflect adaptive diversification Yersinia to their ecological niches. To our knowledge, this is a first investigation of diversity of the porin gene covering the whole genus of the family Enterobacteriaceae. This study demonstrates that recombination and positive selection both contribute to evolution of ompF, but the relative contribution of these evolutionary forces are different among Yersinia species. PMID:21655186

Accessing Suomi NPP OMPS Products through the GES DISC Online Data Services

NASA Technical Reports Server (NTRS)

Johnson, J.; Wei, J.; Gerasimov, I.; Vollmer, Bruce E.

2017-01-01

This presentation will provide an overview of the OMPS products available at the GES DISC archive, as well as demonstrate the various data services provided by the GES DISC. Since the TOMS, SBUV, and EOS Aura (OMI, MLS, HIRDLS) data products are also available from the GES DISC archive, these can be easily accessed and compared with the OMPS data.

Appropriate drinking water treatment processes for organic micropollutants removal based on experimental and model studies - a multi-criteria analysis study.

PubMed

Sudhakaran, Sairam; Lattemann, Sabine; Amy, Gary L

2013-01-01

The presence of organic micropollutants (OMPs), pharmaceuticals and personal care products (PPCPs) in potable water is of great environmental and public health concern. OMPs are included in the priority list of contaminants in United States EPA and European framework directives. Advanced treatment processes such as reverse osmosis, nanofiltration, ozonation and adsorption are the usual industry-recommended processes for OMPs removal, however, natural systems, e.g., riverbank filtration and constructed wetlands, are also potentially efficient options for OMPs removal. In this study, a decision support system (DSS) based on multi-criteria analysis (MCA) was created to compare processes for OMPs removal under various criteria. Multi-criteria analysis (MCA), a transparent and reliable procedure, was adopted. Models were built for both experimental and predicted percent-removals for a range of OMPs reflecting different physicochemical properties. The experimental percent-removals for several processes (riverbank filtration (RBF), ozonation, advanced oxidation, adsorption, reverse osmosis, and nanofiltration) were considered. The predicted percent-removals were taken from validated quantitative structure activity relationship (QSAR) models. Analytical methods to detect OMPs in water are very laborious, thus a modeling approach such as QSAR is an attractive option. A survey among two groups of participants including academics (PhD students and post-doctoral research associates) and industry (managers and operators) representatives was conducted to assign weights for the following criteria: treatability, costs, technical considerations, sustainability and time. The process rankings varied depending on the contaminant species and personal preferences (weights). The results indicated that RBF and oxidation were preferable over adsorption and membranes processes. The results also suggest that the use of a hybrid treatment process, e.g., combining a natural system with an advanced treatment (oxidation) process, may provide benefits for OMPs removal. The proposed DSS can be used as a screening tool for experimental planning or a feasibility study preceding the main treatment system selection and design. It can also be considered as an aid in assessing a multi-barrier approach to remove OMPs. Copyright © 2012 Elsevier B.V. All rights reserved.

OmpR and RcsB abolish temporal and spatial changes in expression of flhD in Escherichia coli biofilm.

PubMed

Samanta, Priyankar; Clark, Emily R; Knutson, Katie; Horne, Shelley M; Prüß, Birgit M

2013-08-02

Biofilms are communities of bacteria that are characterized by specific phenotypes, including an increased resistance towards anti-microbials and the host immune system. This calls for the development of novel biofilm prevention and treatment options to combat infectious disease. In Escherichia coli, numerous global regulators have been implicated in the control of biofilm associated cell surface organelles. These include the flagellar regulator FlhD/FlhC, the osmoregulator EnvZ/OmpR, and the colanic acid activator RcsCDB. Using flow cell technology and fluorescence microscopy, we determined the temporal expression from flhD::gfp, ompR::gfp, and rcsB::gfp in E. coli biofilm, as well as the impact of the negative regulation of flhD by OmpR and RcsB. Spatial gene expression was investigated from flhD::gfp. The temporal gene expression profile for flhD yielded an early peak at 12 h, a minimum of expression at 35 h, and a second increase in expression towards 51 h of biofilm development. In contrast, the ompR profile showed a peak at 35 h. A mutation in ompR abolished time dependence of flhD expression after the initial growth period of 12 h. Intriguingly, rcsB expression did not correlate inversely with flhD expression, yet a mutation in rcsB abolished time dependence of flhD expression as well. Spatially, expression of flhD was highest in the outermost layer of the biofilm in the parent strain. In ompR and rcsB mutants, flhD was expressed throughout the biofilm. Mutations in both, ompR and rcsB increased flhD expression throughout all temporal and spatial experiments. This increase was paralleled by reductions in biofilm amounts at four tested time points. Our data lead to the conclusion that FlhD/FlhC and its regulation by OmpR and RcsB may be our first target mechanism for the development of novel biofilm prevention and treatment techniques.

OmpR and RcsB abolish temporal and spatial changes in expression of flhD in Escherichia coli Biofilm

PubMed Central

2013-01-01

Background Biofilms are communities of bacteria that are characterized by specific phenotypes, including an increased resistance towards anti-microbials and the host immune system. This calls for the development of novel biofilm prevention and treatment options to combat infectious disease. In Escherichia coli, numerous global regulators have been implicated in the control of biofilm associated cell surface organelles. These include the flagellar regulator FlhD/FlhC, the osmoregulator EnvZ/OmpR, and the colanic acid activator RcsCDB. Using flow cell technology and fluorescence microscopy, we determined the temporal expression from flhD::gfp, ompR::gfp, and rcsB::gfp in E. coli biofilm, as well as the impact of the negative regulation of flhD by OmpR and RcsB. Spatial gene expression was investigated from flhD::gfp. Results The temporal gene expression profile for flhD yielded an early peak at 12 h, a minimum of expression at 35 h, and a second increase in expression towards 51 h of biofilm development. In contrast, the ompR profile showed a peak at 35 h. A mutation in ompR abolished time dependence of flhD expression after the initial growth period of 12 h. Intriguingly, rcsB expression did not correlate inversely with flhD expression, yet a mutation in rcsB abolished time dependence of flhD expression as well. Spatially, expression of flhD was highest in the outermost layer of the biofilm in the parent strain. In ompR and rcsB mutants, flhD was expressed throughout the biofilm. Mutations in both, ompR and rcsB increased flhD expression throughout all temporal and spatial experiments. This increase was paralleled by reductions in biofilm amounts at four tested time points. Conclusion Our data lead to the conclusion that FlhD/FlhC and its regulation by OmpR and RcsB may be our first target mechanism for the development of novel biofilm prevention and treatment techniques. PMID:23914787

Expected Performance of Ozone Climate Data Records from Ozone Mapping and Profiler Suite Limb Profiler

NASA Technical Reports Server (NTRS)

Xu, P. Q.; Rault, D. F.; Pawson, S.; Wargan, K.; Bhartia, P. K.

2012-01-01

The Ozone Mapping and Profiler Suite Limb Profiler (OMPS/LP) was launched on board of the Soumi NPP space platform in late October 2011. It provides ozone-profiling capability with high-vertical resolution from 60 Ian to cloud top. In this study, an end-to-end Observing System Simulation Experiment (OSSE) of OMPS/LP ozone is discussed. The OSSE was developed at NASA's Global Modeling and Assimilation Office (GMAO) using the Goddard Earth Observing System (GEOS-5) data assimilation system. The "truth" for this OSSE is built by assimilating MLS profiles and OMI ozone columns, which is known to produce realistic three-dimensional ozone fields in the stratosphere and upper troposphere. OMPS/LP radiances were computed at tangent points computed by an appropriate orbital model. The OMPS/LP forward RT model, Instrument Models (IMs) and EDR retrieval model were introduced and pseudo-observations derived. The resultant synthetic OMPS/LP observations were evaluated against the "truth" and subsequently these observations were assimilated into GEOS-5. Comparison of this assimilated dataset with the "truth" enables comparisons of the likely uncertainties in 3-D analyses of OMPS/LP data. This study demonstrated the assimilation capabilities of OMPS/LP ozone in GEOS-5, with the monthly, zonal mean (O-A) smaller than 0.02ppmv at all levels, the nns(O-A) close to O.lppmv from 100hPa to 0.2hPa; and the mean(O-B) around the 0.02ppmv for all levels. The monthly zonal mean analysis generally agrees to within 2% of the truth, with larger differences of 2-4% (0.1-0.2ppmv) around 10hPa close to North Pole and in the tropical tropopause region, where the difference is above 20% due to the very low ozone concentrations. These OSSEs demonstrated that, within a single data assimilation system and the assumption that assimilated MLS observations provide a true rendition of the stratosphere, the OMPS/LP ozone data are likely to produce accurate analyses through much of the stratosphere. Additional studies are needed to evaluate the importance of the extremely high vertical resolution of the OMPS/LP ozone data

Comparison of Aerobic Preservation by Venous Systemic Oxygen Persufflation or Oxygenated Machine Perfusion of Warm-Ischemia-Damaged Porcine Kidneys.

PubMed

Kalenski, Julia; Mancina, Elina; Paschenda, Pascal; Beckers, Christian; Bleilevens, Christian; Tóthová, Ľubomíra; Boor, Peter; Gross, Dominik; Tolba, René H; Doorschodt, Benedict M

2016-01-01

The global shortage of donor organs for transplantation has necessitated the expansion of the organ pool through increased use of organs from less ideal donors. Venous systemic oxygen persufflation (VSOP) and oxygenated machine perfusion (OMP) have previously demonstrated beneficial results compared to cold storage (CS) in the preservation of warm-ischemia-damaged kidney grafts. The aim of this study was to compare the efficacy of VSOP and OMP for the preservation of warm-ischemia-damaged porcine kidneys using the recently introduced Ecosol preservation solution compared to CS using Ecosol or histidine-tryptophan-ketoglutarate solution (HTK). Kidneys from German Landrace pigs (n = 5/group) were retrieved and washed out with either Ecosol or HTK after 45 min of clamping of the renal pedicle. As controls, kidneys without warm ischemia, cold stored for 24 h in HTK, were employed. Following 24 h of preservation by VSOP, OMP, CS-Ecosol, or CS-HTK, renal function and damage were assessed during 1 h using the isolated perfused porcine kidney model. During reperfusion, urine production was significantly higher in the VSOP and OMP groups than in the CS-HTK group; however, only VSOP could demonstrate lower urine protein concentrations and fractional excretion of sodium, which did not differ from the non-warm-ischemia-damaged control group. VSOP, CS-Ecosol, and controls showed better maintenance of the acid-base balance than CS-HTK. Reduced lipid peroxidation, as reflected in postreperfusion tissue thiobarbituric acid-reactive substance levels, was observed in the VSOP group compared to the OMP group, and the VSOP and CS-Ecosol groups had concentrations similar to the controls. The ratio of reduced to oxidized glutathione was higher in the VSOP, OMP, and CS-Ecosol groups than in the CS-HTK group and controls, with a higher ratio in the VSOP than in the OMP group. VSOP was associated with mitigation of oxidative stress in comparison to OMP and CS. Preservation of warm-ischemia-damaged porcine kidneys by VSOP was improved compared to OMP and CS, and was comparable to preservation of non-warm-ischemia-damaged cold-stored kidneys. © 2016 S. Karger AG, Basel.

Periplasmic quality control in biogenesis of outer membrane proteins.

PubMed

Lyu, Zhi Xin; Zhao, Xin Sheng

2015-04-01

The β-barrel outer membrane proteins (OMPs) are integral membrane proteins that reside in the outer membrane of Gram-negative bacteria and perform a diverse range of biological functions. Synthesized in the cytoplasm, OMPs must be transported across the inner membrane and through the periplasmic space before they are assembled in the outer membrane. In Escherichia coli, Skp, SurA and DegP are the most prominent factors identified to guide OMPs across the periplasm and to play the role of quality control. Although extensive genetic and biochemical analyses have revealed many basic functions of these periplasmic proteins, the mechanism of their collaboration in assisting the folding and insertion of OMPs is much less understood. Recently, biophysical approaches have shed light on the identification of the intricate network. In the present review, we summarize recent advances in the characterization of these key factors, with a special emphasis on the multifunctional protein DegP. In addition, we present our proposed model on the periplasmic quality control in biogenesis of OMPs.

Recognition of foreign oviposition-marking pheromone in a multi-trophic context

NASA Astrophysics Data System (ADS)

Stelinski, L. L.; Rodriguez-Saona, C.; Meyer, W. L.

2009-05-01

Both phytophagous and parasitic insects deposit oviposition-marking pheromones (OMPs) following oviposition that function to inform conspecifics of a previously utilized host of reduced suitability. The blueberry maggot fly, Rhagoletis mendax Curran (Diptera: Tephritidae), deposits eggs individually into blueberries and then marks the fruit surface with an OMP which reduces acceptance of fruit for oviposition by conspecifics. Diachasma alloeum (Muesebeck) (Hymenoptera: Braconidae) is a parasitic wasp attacking larval R. mendax which also deposits an OMP, signaling conspecifics of a wasp-occupied host. Behavioral studies were conducted testing the hypothesis that the OMP of the parasitic wasp modifies the oviposition behavior of its host fly. In this study, we show that the OMP of D. alloeum is recognized by R. mendax, and female flies will reject wasp-marked fruit for oviposition. Thus, we present a rare demonstration of pheromonal recognition between animals occupying different taxonomic orders and trophic levels. This chemical eavesdropping may enhance the ability of the fly to avoid fruit unsuitable for larval development.

OMP Peptides Activate the DegS Stress-Sensor Protease by a Relief of Inhibition Mechanism

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sohn, Jungsan; Grant, Robert A.; Sauer, Robert T.

2010-03-19

In the E. coli periplasm, C-terminal peptides of misfolded outer-membrane porins (OMPs) bind to the PDZ domains of the trimeric DegS protease, triggering cleavage of a transmembrane regulator and transcriptional activation of stress genes. We show that an active-site DegS mutation partially bypasses the requirement for peptide activation and acts synergistically with mutations that disrupt contacts between the protease and PDZ domains. Biochemical results support an allosteric model, in which these mutations, active-site modification, and peptide/substrate binding act in concert to stabilize proteolytically active DegS. Cocrystal structures of DegS in complex with different OMP peptides reveal activation of the proteasemore » domain with varied conformations of the PDZ domain and without specific contacts from the bound OMP peptide. Taken together, these results indicate that the binding of OMP peptides activates proteolysis principally by relieving inhibitory contacts between the PDZ domain and the protease domain of DegS.« less

USC orthogonal multiprocessor for image processing with neural networks

NASA Astrophysics Data System (ADS)

Hwang, Kai; Panda, Dhabaleswar K.; Haddadi, Navid

1990-07-01

This paper presents the architectural features and imaging applications of the Orthogonal MultiProcessor (OMP) system, which is under construction at the University of Southern California with research funding from NSF and assistance from several industrial partners. The prototype OMP is being built with 16 Intel i860 RISC microprocessors and 256 parallel memory modules using custom-designed spanning buses, which are 2-D interleaved and orthogonally accessed without conflicts. The 16-processor OMP prototype is targeted to achieve 430 MIPS and 600 Mflops, which have been verified by simulation experiments based on the design parameters used. The prototype OMP machine will be initially applied for image processing, computer vision, and neural network simulation applications. We summarize important vision and imaging algorithms that can be restructured with neural network models. These algorithms can efficiently run on the OMP hardware with linear speedup. The ultimate goal is to develop a high-performance Visual Computer (Viscom) for integrated low- and high-level image processing and vision tasks.

OmpA: A Flexible Clamp for Bacterial Cell Wall Attachment.

PubMed

Samsudin, Firdaus; Ortiz-Suarez, Maite L; Piggot, Thomas J; Bond, Peter J; Khalid, Syma

2016-12-06

The envelope of Gram-negative bacteria is highly complex, containing separate outer and inner membranes and an intervening periplasmic space encompassing a peptidoglycan (PGN) cell wall. The PGN scaffold is anchored non-covalently to the outer membrane via globular OmpA-like domains of various proteins. We report atomically detailed simulations of PGN bound to OmpA in three different states, including the isolated C-terminal domain (CTD), the full-length monomer, or the complete full-length dimeric form. Comparative analysis of dynamics of OmpA CTD from different bacteria helped to identify a conserved PGN-binding mode. The dynamics of full-length OmpA, embedded within a realistic representation of the outer membrane containing full-rough (Ra) lipopolysaccharide, phospholipids, and cardiolipin, suggested how the protein may provide flexible mechanical support to the cell wall. An accurate model of the heterogeneous bacterial cell envelope should facilitate future efforts to develop antibacterial agents. Copyright © 2016 Elsevier Ltd. All rights reserved.

Sequencing of the Chlamydophila psittaci ompA Gene Reveals a New Genotype, E/B, and the Need for a Rapid Discriminatory Genotyping Method

PubMed Central

Geens, Tom; Desplanques, Ann; Van Loock, Marnix; Bönner, Brigitte M.; Kaleta, Erhard F.; Magnino, Simone; Andersen, Arthur A.; Everett, Karin D. E.; Vanrompay, Daisy

2005-01-01

Twenty-one avian Chlamydophila psittaci isolates from different European countries were characterized using ompA restriction fragment length polymorphism, ompA sequencing, and major outer membrane protein serotyping. Results reveal the presence of a new genotype, E/B, in several European countries and stress the need for a discriminatory rapid genotyping method. PMID:15872282

Study the Expression of ompf Gene in Esherichia coli Mutants.

PubMed

Jaktaji, R Pourahmad; Heidari, F

2013-09-01

The outer membrane porin proteins are the major factors in controlling the permeability of cell membrane. OmpF is an example of porin proteins in Esherichia coli. In normal growth condition a large amount of this protein is synthesised, but under stress condition, such as the presence of antibiotics in environment its expression is decreased inhibiting the entrance of antibiotics into cell. The expression of ompF is inhibited by antisense RNA transcribed from micF. In normal condition the expression of micF is low, but in the presence of antibiotics its expression is increased and causes multiple resistances to irrelevant antibiotics. The aims of this research were to study first, the intactness of micF and then quantify the expression of ompF in ciprofloxacin and tetracycline resistant mutants of E. coli. For this purpose the 5' end of micF was amplified and then sequenced. None of these mutants except one and its clone has a mutation in this gene. Then the relative expression of ompF in these mutants was quantified by real time PCR. There was no significant difference between ompF transcription of mutants and wild type strain. Based on this study and previous study it is concluded that low to intermediate levels of resistance to ciprofloxacin and tetracycline does not decrease ompF transcription.

Ducks as a potential reservoir for Pasteurella multocida infection detected using a new rOmpH-based ELISA.

PubMed

Liu, Rongchang; Chen, Cuiteng; Cheng, Longfei; Lu, Ronghui; Fu, Guanghua; Shi, Shaohua; Chen, Hongmei; Wan, Chunhe; Lin, Jiansheng; Fu, Qiuling; Huang, Yu

2017-07-28

Pasteurella multocida is an important pathogen of numerous domestic poultry and wild animals and is associated with a variety of diseases including fowl cholera. The aim of this study was to develop an indirect enzyme-linked immunosorbent assay (ELISA) based on recombinant outer-membrane protein H (rOmpH) for detection of anti-P. multocida antibodies in serum to determine their prevalence in Chinese ducks. The P. multocida ompH gene was cloned into pET32a, and rOmpH was expressed in Escherichia coli BL21 (DE3). Western blotting revealed that purified rOmpH was recognized by duck antisera against P. multocida, and an indirect ELISA was established. During analysis of serum samples (n=115) from ducks, the rOmpH ELISA showed 95.0% specificity, 100% sensitivity and a 92.0% κ coefficient (95% confidence interval 0.844-0.997) as compared with a microtiter agglutination test. Among 165 randomly selected serum samples, which were collected in 2015 and originated from six duck farms across Fujian Province, China, anti-P. multocida antibodies were detected in 22.42% of apparently healthy ducks, including 25 of 90 sheldrakes (27.8%), eight of 50 Peking ducks (16.0%) and four of 25 Muscovy ducks (16%). Overall, the data suggest that rOmpH is a suitable candidate antigen for the development of an indirect ELISA for detection of P. multocida in ducks; moreover, our results showed that ducks could serve as a potential reservoir for P. multocida infection.

Insights into PG-binding, conformational change, and dimerization of the OmpA C-terminal domains from Salmonella enterica serovar Typhimurium and Borrelia burgdorferi: Characterization of OmpA C-Terminal Domain

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tan, Kemin; Deatherage Kaiser, Brooke L.; Wu, Ruiying

S. Typhimurium can induce both humoral and cell-mediated responses when establishing itself in the host. These responses are primarily stimulated against the lipopolysaccharide and major outer membrane (OM) proteins. OmpA is one of these major OM proteins. It comprises a N-terminal eight-stranded b-barrel trans membrane domain and a C-terminal domain (OmpACTD). The OmpACTD and its homologs are believed to bind to peptidoglycan (PG) within the periplasm, maintaining bacterial osmotic homeostasis and modulating the permeability and integrity of the OM. Here we present the first crystal structures of the OmpACTD from two pathogens: S. Typhimurium (STOmpACTD) in open and closed formsmore » and causative agent of Lyme Disease Borrelia burgdorferi (BbOmpACTD), in closed form. In the open form of STOmpACTD, an aspartic acid residue from a long b2-a3 loop points into the binding pocket, suggesting that an anion group such as a carboxylate group from PG is favored at the binding site. In the closed form of STOmpACTD and in the structure of BbOmpACTD, a sulfate group from the crystallization buffer is tightly bound at the binding site. The differences between the closed and open forms of STOmpACTD, suggest a large conformational change that includes an extension of a3 helix by ordering a part of b2-a3 loop. We propose that the sulfate anion observed in these structures mimics the carboxylate group of PG when bound to STOmpACTD suggesting PG-anchoring mechanism. In addition, the binding of PG or a ligand mimic may enhance dimerization of STOmpACTD, or possibly that of full length STOmpA.« less

Identifying the location of the OMP separatrix in DIII-D using power accounting

DOE PAGES

Stangeby, Peter C.; Canik, John M.; Elder, J. D.; ...

2015-08-07

In order to identify reliable scalings for the scrape-off layer (SOL) power width it is necessary to know the location of the separatrix in divertor tokamaks as accurately as possible, specifically its location at the outside midplane (OMP) the standard reference location. Two methods are described which use power accounting to improve the accuracy of identifying the location of the OMP separatrix. The first uses the infrared-measured deposited power profile at the outer target as the primary input, the 'more » $$P_{{\\rm SOL}}^{{\\rm exhaust}}$$ method'. The second uses the measured power input to the SOL, obtained by subtracting the power radiated from inside the separatrix from the total heating power, the ' $$P_{{\\rm SOL}}^{{\\rm input}}$$ method'. Furthermore, these two power accounting methods are illustrated with the examples of 21 H-mode DIII-D discharges. High spatial resolution Thomson scattering measured profiles of ne and Te for the main SOL near the OMP are also used as primary input to the analysis; only between-edge localized mode data are used here. The Thomson profiles are used to calculate the electron parallel conducted heat flux profiles which are then matched to the measured $$P_{{\\rm SOL}}^{{\\rm exhaust}}$$ and $$P_{{\\rm SOL}}^{{\\rm input}}$$ by adjusting the location of the OMP separatrix relative to that of the Thomson data. For these attached discharges, it is found that the values of $$R_{{\\rm sep}}^{{\\rm omp}}$$ given by the two power accounting methods agree to within ~1 mm of each other and also to within ~1 mm of the values given by the 'standard DIII-D method' described by Porter et al (1998 Phys. Plasmas 5 1410). Lastly, the shifted $$R_{{\\rm sep}}^{{\\rm omp}}$$ results in only modest changes to the values of ne and Te at the OMP separatrix relative to the 'standard' values, increasing $$n_{{\\rm e}}^{{\\rm sep}}$$ by 8% and $$T_{{\\rm e}}^{{\\rm sep}}$$ by 20%.« less

Clinical evidence for orphan medicinal products-a cause for concern?

PubMed

Picavet, Eline; Cassiman, David; Hollak, Carla E; Maertens, Johan A; Simoens, Steven

2013-10-16

The difficulties associated with organising clinical studies for orphan medicinal products (OMPs) are plentiful. Recent debate on the long-term effectiveness of some OMPs, led us to question whether the initial standards for clinical evidence for OMPs, set by the European Medicines Agency (EMA) at the time of marketing authorization, are too low. Therefore, the aim of this study was to quantitatively evaluate the characteristics and quality of clinical evidence that is presented for OMPs to obtain marketing authorization in Europe, using the new and validated COMPASS tool. We quantitatively assessed the characteristics and quality of clinical evidence of the pivotal studies of 64 OMPs as described in the European Public Assessment Report and/or the Scientific Discussion document prepared by the Committee for Human Medicinal Products of the EMA. The 64 OMPs were altogether authorized for 78 orphan indications, for which 117 studies were identified as 'pivotal' or 'main' studies. In approximately two thirds of the studies, the allocation was randomized (64.8%) and a control arm was used (68.5%). Half of the studies applied some type of blinding. Only a minority (26.9%) of the studies included a Quality-of-Life (QoL) related endpoint, of which a third claim an improvement in QoL. Upon analyzing the quality of reporting, we found that some aspects (i.e. the endpoints, the sampling criteria, and the interventions) are well described, whereas other items (i.e. a description of the patients and of potential biases) are not reported for all studies. In conclusion, the pivotal studies that are the basis for marketing authorization of OMPs are a cause for concern, as they exhibit methodological flaws i.e. the lack of QoL-related endpoints as outcome, lack of blinding in the study design and the use of surrogate endpoints. Additionally, there are shortcomings in the reporting of those studies that complicate the interpretation. A more demanding regulatory process for OMPs is needed to guide evidence-based clinical decision-making.

Recombinant Hsp33 and OmpC protein can serve as promising divalent vaccine with protection against Vibrio anguillarum and Edwardsiella tarda in flounder (Paralichthys olivaceus).

PubMed

Xing, Jing; Li, Pengwei; Tang, Xiaoqian; Zhan, Wenbin

2018-03-01

Vibrio anguillarum and Edwardsiella tarda are severe aquaculture pathogens shared similar epidemiological characteristics and susceptible to flounder (Paralichthys olivaceus). In our previous studies, recombinant(r) protein heat shock protein 33 (rHsp33) from V. anguillarum and outer membrane protein C (rOmpC) from E. tarda were proved to have protection against V. anguillarum and E. tarda, respectively. In this paper, the cross protection of rHsp33 against E. tarda and rOmpC against V. anguillarum, and the protection of divalent vaccine candidate (rHsp33 + rOmpC, rHC) against both V. anguillarum and E. tarda were evaluated. RHC, rHsp33, and rOmpC were vaccinated to flounder, respectively, and the percentages of surface immunoglobulin-positive (sIg+) cells in peripheral blood lymphocytes (PBLs), serum IgM, specific antibodies against V. anguillarum or E. tarda, specific antibodies against rHsp33, rOmpC or rHC, the expression of immune-related genes and relative percent survival (RPS) against V. anguillarum or E. tarda were measured. The results showed that: RHC could induced the enhancement of sIg + cells and high levels of specific antibodies against both V. anguillarm and E. tarda; Also a significant increase of specific antibodies against rHsp33, rOmpC or rHC, and up-regulation of gene expression of CD3, CD4-1, CD4-2, CD8α, CD8β and IgM in spleen, head-kidney, and hindgut, RPS of 70 ± 3.45% against V. anguillarum and 60 ± 1.48% against E. tarda, respectively. In addition, rHsp33 induced specific antibodies against E. tarda and rOmpC, and had a RPS of 43.3 ± 3.73% against E. tarda; rOmpC could evoke specific antibodies against V. anguillarum and rHsp33, and had a RPS of 44 ± 1.27% against V. anguillarm; The results demonstrated that there was cross protection of rHsp33 against E. tarda and rOmpC against V. anguillarum, rHC as a divalent vaccine can induce significant immune response and efficient protection against both E. tarda and V. anguillarum in flounder. Copyright © 2018 Elsevier Ltd. All rights reserved.

Analysis of the SDS-PAGE patterns of outer membrane proteins from Escherichia coli strains that have lost the ability to form K1 antigen and varied in the susceptibility to normal human serum.

PubMed

Cisowska, Agnieszka; Bugla-Płoskońska, Gabriela

2014-01-01

We used SDS-polyacrylamide gel electrophoresis to investigate the outer membrane proteins (OMPs) band composition of 19 Escherichia coli K1 strains that have spontaneously lost the ability to form K1 polysaccharide capsule (E. coli K1-) and demonstrated different degrees of susceptibility to the bactericidal action of normal human serum. Presented results showed that there were differences between E. coli K1- strains in OMPs expressing capacity. The analysis performed on OMPs has not revealed a direct association between the different OMPs band composition and the susceptibility of these strains to the serum.

Master Amino acid Pattern as sole and total substitute for dietary proteins during a weight-loss diet to achieve the body's nitrogen balance equilibrium.

PubMed

Lucà-Moretti, M; Grandi, A; Lucà, E; Muratori, G; Nofroni, M G; Mucci, M P; Gambetta, P; Stimolo, R; Drago, P; Giudice, G; Tamburlin, N; Karbalai, M; Valente, C; Moras, G

2003-01-01

Results of this multicentric study have shown that by giving Master Amino acid Pattern (MAP) as a sole and total substitute of dietary proteins to 500 overweight participants undergoing the American Nutrition Clinics/Overweight Management Program (ANC/OMP), the participants' body nitrogen balance could be maintained in equilibrium with essentially no calories (MAP 1 g=0.04 kcal), thereby preserving the body's structural and functional proteins, eliminating excessive water retention from the interstitial compartment, and preventing the sudden weight increase after study conclusion commonly known as the yo-yo effect. Study results have shown that the use of MAP, in conjunction with the ANC/OMP regimen, has proven to be safe and effective by preventing those adverse effects associated with a negative nitrogen balance, such as oversized or flabby tissue, stretch marks, the sagging of breast tissue, increased hair loss, faded hair color, and fragile or brittle nails. Also prevented were those anomalies commonly associated with weight-loss diets, such as hunger, weakness, headache caused by ketosis, constipation, and decreased libido. The use of MAP in conjunction with the ANC/OMP also allowed for mean weight loss of 2.5 kg (5.5 lb) per week, achieved through reduction of excessive fat tissue and elimination of excessive water retention from the interstitial compartment.

Continuation of long-term global SO2 pollution monitoring from OMI to OMPS

NASA Astrophysics Data System (ADS)

Zhang, Y.; Li, C.; Krotkov, N. A.; Joiner, J.

2016-12-01

In the past 12+ years, Ozone Monitoring Instrument (OMI) on board NASA EOS Aura satellite has pioneered the first high-resolution global SO2 pollution monitoring, which enabled new studies of atmospheric chemistry and applications for air quality management. Such long-term SO2 record will be continued with other satellite instruments, i.e., the Ozone Mapping and Profiler Suite (OMPS) Nadir Mapper on board NASA-NOAA Suomi National Polar-orbiting Partnership (S-NPP) satellite and the follow up JPSS series satellites. In this presentation, we demonstrate the first comparison between OMI and OMPS SO2 retrievals from the OMI operational SO2 algorithm, which is our state-of-the-art principal component analysis (PCA) approach. The PCA technique does not use any sort of "soft calibration" corrections required in concurrent satellite SO2 algorithms and enables seamless merging of different satellite datasets. We demonstrate a very good consistency of the retrievals from OMI and OMPS. Four full years of OMI and OMPS SO2 retrievals during 2012-2015 have been analyzed over some of the world's most polluted regions: eastern China, Mexico, and South Africa. In general, the comparisons show high correlations (r =0.79-0.96) of SO2 mass between the two instruments on a daily basis and less than unity regression slopes (0.76-0.97) indicating slightly lower OMPS SO2 mass as compared with OMI. The annual averaged SO2 loading difference between OMI and OMPS is negligible (< 0.03 Dobson Unit (DU)) over South Africa and up to 0.1 DU over eastern China). We also found a very good correlation (r=0.92-0.97) between the spatial distributions of the annual mean SO2 over the three regions. The two instruments also show generally good agreement in terms of the daily spatial distribution in SO2. For example, over the Mexico region for 82% of the days, the two instruments have a spatial correlation coefficient of 0.6 or better. Such consistent retrievals were achieved without any explicit adjustment to OMI or OMPS radiance data. We will further improve the retrieval agreement in the next versions of the OMI and OMPS retrievals by applying a more comprehensive Jacobian lookup table that properly accounts for different observation conditions.

The toxR Gene of Vibrio (Listonella) anguillarum Controls Expression of the Major Outer Membrane Proteins but Not Virulence in a Natural Host Model

PubMed Central

Okuda, Jun; Nakai, Toshihiro; Chang, Park Se; Oh, Takanori; Nishino, Takeshi; Koitabashi, Tsutomu; Nishibuchi, Mitsuaki

2001-01-01

To examine the hypothesis that the ancestral role of the toxR gene in the family Vibrionaceae is control of the expression of outer membrane protein (OMP)-encoding genes for adaptation to environmental change, we investigated the role of the toxR gene in Vibrio anguillarum, an important fish pathogen. The toxR gene of V. angullarum (Va-toxR) was cloned from strain PT-87050 isolated from diseased ayu (Plecoglossus altivelis), and the sequence was analyzed. The toxR sequence was 63 to 51% identical to those reported for other species of the family Vibrionaceae. Distribution of the Va-toxR gene sequence in V. anguillarum strains of various serotypes was confirmed by using DNA probe and PCR methods. An isogenic toxR mutant of V. anguillarum PT-24, isolated from diseased ayu, was constructed by using an allelic exchange method. The wild-type strain and the toxR mutant did not differ in the ability to produce a protease(s) and a hemolysin(s) or in pathogenicity for ayu when examined by the intramuscular injection and immersion methods. A 35-kDa major OMP was not produced by the toxR mutant. However, a 46-kDa OMP was hardly detected in the wild-type strain but was produced as the major OMP by the toxR mutant. For the toxR mutant, the MICs of two β-lactam antibiotics were higher and the minimum bactericidal concentration of sodium dodecyl sulfate was lower than for the wild-type strain. Analysis of the N-terminal amino acid sequences of the 35- and 46-kDa OMPs indicated that these proteins are the porin-like OMPs and are related to the toxR-regulated major OMPs of the family Vibrionaceae. The results indicate that the toxR gene is not involved in virulence expression in V. anguillarum PT-24 and that toxR regulation of major OMPs is universal in the family Vibrionaceae. These results support the hypothesis that the ancestral role of the toxR gene is regulation of OMP gene expression and that only in some Vibrio species has ToxR been appropriated for the regulation of a virulence gene(s). PMID:11553547

Distribution of the ompA-types among ruminant and swine pneumonic strains of Pasteurella multocida exhibiting various cap-locus and toxA patterns.

PubMed

Vougidou, C; Sandalakis, V; Psaroulaki, A; Siarkou, V; Petridou, E; Ekateriniadou, L

2015-05-01

Pasteurella multocida is an important pathogen in food-producing animals and numerous virulence genes have been identified in an attempt to elucidate the pathogenesis of pasteurellosis. Currently, some of these genes including the capsule biosynthesis genes, the toxA and the OMPs-encoding genes have been suggested as epidemiological markers. However, the number of studies concerning ruminant isolates is limited, while, no attempt has ever been made to investigate the existence of ompA sequence diversity among P. multocida isolates. The aim of the present study was the comparative analysis of 144 P. multocida pneumonic isolates obtained from sheep, goats, cattle and pigs by determining the distribution of the ompA-types in conjunction with the cap-locus and toxA patterns. The ompA genotypes of the isolates were determined using both a PCR-RFLP method and DNA sequence analysis. The most prevalent capsule biosynthesis gene among the isolates was capA (86.1%); a noticeable, however, rate of capD-positive isolates (38.6%) was found among the ovine isolates that had been associated primarily with the capsule type A in the past. Moreover, an unexpectedly high percentage of toxA-positive pneumonic isolates was noticed among small ruminants (93.2% and 85.7% in sheep and goats, respectively), indicating an important epidemiological role of toxigenic P. multocida for these species. Despite their great heterogeneity, certain ompA-genotypes were associated with specific host species, showing evidence of a host preference. The OmpA-based PCR-RFLP method developed proved to be a valuable tool in typing P. multocida strains. Copyright © 2015 Elsevier GmbH. All rights reserved.

Immunological characteristics of outer membrane protein omp31 of goat Brucella and its monoclonal antibody.

PubMed

Zheng, W Y; Wang, Y; Zhang, Z C; Yan, F

2015-10-05

We examined the immunological characteristics of outer membrane protein omp31 of goat Brucella and its monoclonal antibody. Genomic DNA from the M5 strain of goat Brucella was amplified by polymerase chain reaction and cloned into the prokaryotic expression vector pGEX-4T-1. The expression and immunological characteristics of the fusion protein GST-omp31 were subjected to preliminary western blot detection with goat Brucella rabbit immune serum. The Brucella immunized BALB/c mouse serum was detected using purified protein. The high-potency mouse splenocytes and myeloma Sp2/0 cells were fused. Positive clones were screened by enzyme-linked immunosorbent assay to establish a hybridoma cell line. Mice were inoculated intraperitoneally with hybridoma cells to prepare ascites. The mAb was purified using the n-caprylic acid-ammonium sulfate method. The characteristics of mAb were examined using western blotting and enzyme-linked immunosorbent assay. A 680-base pair band was observed after polymerase chain reaction. Enzyme digestion identification and sequencing showed that the pGEX-4T-1-omp31 prokaryotic expression vector was successfully established; a target band of approximately 57 kDa with an apparent molecular weight consistent with the size of the target fusion protein. At 25°C, the expression of soluble expression increased significantly; the fusion protein GST-omp31 was detected by western blotting. Anti-omp31 protein mAb was obtained from 2 strains of Brucella. The antibody showed strong specificity and sensitivity and did not cross-react with Escherichia coli, Staphylococcus aureus, Bacillus subtilis, Mycobacterium tuberculosis, or Bacillus pyocyaneus. The pGEX-4T-1-omp31 prokaryotic expression vector was successfully established and showed good immunogenicity. The antibody also showed strong specificity and good sensitivity.

Automated and Assistive Tools for Accelerated Code migration of Scientific Computing on to Heterogeneous MultiCore Systems

DTIC Science & Technology

2017-04-13

modelling code, a parallel benchmark , and a communication avoiding version of the QR algorithm. Further, several improvements to the OmpSs model were...movement; and a port of the dynamic load balancing library to OmpSs. Finally, several updates to the tools infrastructure were accomplished, including: an...OmpSs: a basic algorithm on image processing applications, a mini application representative of an ocean modelling code, a parallel benchmark , and a

User interface issues in supporting human-computer integrated scheduling

NASA Technical Reports Server (NTRS)

Cooper, Lynne P.; Biefeld, Eric W.

1991-01-01

Explored here is the user interface problems encountered with the Operations Missions Planner (OMP) project at the Jet Propulsion Laboratory (JPL). OMP uses a unique iterative approach to planning that places additional requirements on the user interface, particularly to support system development and maintenance. These requirements are necessary to support the concepts of heuristically controlled search, in-progress assessment, and iterative refinement of the schedule. The techniques used to address the OMP interface needs are given.

Integrating powdered activated carbon into wastewater tertiary filter for micro-pollutant removal.

PubMed

Hu, Jingyi; Aarts, Annelies; Shang, Ran; Heijman, Bas; Rietveld, Luuk

2016-07-15

Integrating powdered activated carbon (PAC) into wastewater tertiary treatment is a promising technology to reduce organic micro-pollutant (OMP) discharge into the receiving waters. To take advantage of the existing tertiary filter, PAC was pre-embedded inside the filter bed acting as a fixed-bed adsorber. The pre-embedding (i.e. immobilization) of PAC was realized by direct dosing a PAC solution on the filter top, which was then promoted to penetrate into the filter media by a down-flow of tap water. In order to examine the effectiveness of this PAC pre-embedded filter towards OMP removal, batch adsorption tests, representing PAC contact reactor (with the same PAC mass-to-treated water volume ratio as in the PAC pre-embedded filter) were performed as references. Moreover, as a conventional dosing option, PAC was dosed continuously with the filter influent (i.e. the wastewater secondary effluent with the investigated OMPs). Comparative results confirmed a higher OMP removal efficiency associated with the PAC pre-embedded filter, as compared to the batch system with a practical PAC residence time. Furthermore, over a filtration period of 10 h (approximating a realistic filtration cycle for tertiary filters), the continuous dosing approach resulted in less OMP removal. Therefore, it was concluded that the pre-embedding approach can be preferentially considered when integrating PAC into the wastewater tertiary treatment for OMP elimination. Copyright © 2016 Elsevier Ltd. All rights reserved.

A Supercomplex Spanning the Inner and Outer Membranes Mediates the Biogenesis of β-Barrel Outer Membrane Proteins in Bacteria.

PubMed

Wang, Yan; Wang, Rui; Jin, Feng; Liu, Yang; Yu, Jiayu; Fu, Xinmiao; Chang, Zengyi

2016-08-05

β-barrel outer membrane proteins (OMPs) are ubiquitously present in Gram-negative bacteria, mitochondria and chloroplasts, and function in a variety of biological processes. The mechanism by which the hydrophobic nascent β-barrel OMPs are transported through the hydrophilic periplasmic space in bacterial cells remains elusive. Here, mainly via unnatural amino acid-mediated in vivo photo-crosslinking studies, we revealed that the primary periplasmic chaperone SurA interacts with nascent β-barrel OMPs largely via its N-domain but with β-barrel assembly machine protein BamA mainly via its satellite P2 domain, and that the nascent β-barrel OMPs interact with SurA via their N- and C-terminal regions. Additionally, via dual in vivo photo-crosslinking, we demonstrated the formation of a ternary complex involving β-barrel OMP, SurA, and BamA in cells. More importantly, we found that a supercomplex spanning the inner and outer membranes and involving the BamA, BamB, SurA, PpiD, SecY, SecE, and SecA proteins appears to exist in living cells, as revealed by a combined analyses of sucrose-gradient ultra-centrifugation, Blue native PAGE and mass spectrometry. We propose that this supercomplex integrates the translocation, transportation, and membrane insertion events for β-barrel OMP biogenesis. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

Study the Expression of ompf Gene in Esherichia coli Mutants

PubMed Central

Jaktaji, R. Pourahmad; Heidari, F.

2013-01-01

The outer membrane porin proteins are the major factors in controlling the permeability of cell membrane. OmpF is an example of porin proteins in Esherichia coli. In normal growth condition a large amount of this protein is synthesised, but under stress condition, such as the presence of antibiotics in environment its expression is decreased inhibiting the entrance of antibiotics into cell. The expression of ompF is inhibited by antisense RNA transcribed from micF. In normal condition the expression of micF is low, but in the presence of antibiotics its expression is increased and causes multiple resistances to irrelevant antibiotics. The aims of this research were to study first, the intactness of micF and then quantify the expression of ompF in ciprofloxacin and tetracycline resistant mutants of E. coli. For this purpose the 5’ end of micF was amplified and then sequenced. None of these mutants except one and its clone has a mutation in this gene. Then the relative expression of ompF in these mutants was quantified by real time PCR. There was no significant difference between ompF transcription of mutants and wild type strain. Based on this study and previous study it is concluded that low to intermediate levels of resistance to ciprofloxacin and tetracycline does not decrease ompF transcription. PMID:24403654

A Supercomplex Spanning the Inner and Outer Membranes Mediates the Biogenesis of β-Barrel Outer Membrane Proteins in Bacteria*

PubMed Central

Wang, Yan; Wang, Rui; Jin, Feng; Liu, Yang; Yu, Jiayu; Fu, Xinmiao; Chang, Zengyi

2016-01-01

β-barrel outer membrane proteins (OMPs) are ubiquitously present in Gram-negative bacteria, mitochondria and chloroplasts, and function in a variety of biological processes. The mechanism by which the hydrophobic nascent β-barrel OMPs are transported through the hydrophilic periplasmic space in bacterial cells remains elusive. Here, mainly via unnatural amino acid-mediated in vivo photo-crosslinking studies, we revealed that the primary periplasmic chaperone SurA interacts with nascent β-barrel OMPs largely via its N-domain but with β-barrel assembly machine protein BamA mainly via its satellite P2 domain, and that the nascent β-barrel OMPs interact with SurA via their N- and C-terminal regions. Additionally, via dual in vivo photo-crosslinking, we demonstrated the formation of a ternary complex involving β-barrel OMP, SurA, and BamA in cells. More importantly, we found that a supercomplex spanning the inner and outer membranes and involving the BamA, BamB, SurA, PpiD, SecY, SecE, and SecA proteins appears to exist in living cells, as revealed by a combined analyses of sucrose-gradient ultra-centrifugation, Blue native PAGE and mass spectrometry. We propose that this supercomplex integrates the translocation, transportation, and membrane insertion events for β-barrel OMP biogenesis. PMID:27298319

Brucella ovis PA mutants for outer membrane proteins Omp10, Omp19, SP41, and BepC are not altered in their virulence and outer membrane properties.

PubMed

Sidhu-Muñoz, Rebeca S; Sancho, Pilar; Vizcaíno, Nieves

2016-04-15

Mutants in several genes have been obtained on the genetic background of virulent rough (lacking O-polysaccharide) Brucella ovis PA. The target genes encode outer membrane proteins previously associated with the virulence of smooth (bearing O-polysaccharide chains in the lipopolysaccharide) Brucella strains. Multiple attempts to delete omp16, coding for a homologue to peptidoglycan-associated lipoproteins, were unsuccessful, which suggests that Omp16 is probably essential for in vitro survival of B. ovis PA. Single deletion of omp10 or omp19-that encode two other outer membrane lipoproteins--was achieved, but the simultaneous removal of both genes failed, suggesting an essential complementary function between both proteins. Two other deletion mutants, defective in the Tol-C-homologue BepC or in the SP41 adhesin, were also obtained. Surprisingly when compared to previous results obtained with smooth Brucella, none of the B. ovis mutants showed attenuation in the virulence, either in the mouse model or in cellular models of professional and non-professional phagocytes. Additionally, and in contrast to the observations reported with smooth Brucella strains, several properties related to the outer membrane remained almost unaltered. These results evidence new distinctive traits between naturally rough B. ovis and smooth brucellae. Copyright © 2016 Elsevier B.V. All rights reserved.

Identification of specific posttranslational O-mycoloylations mediating protein targeting to the mycomembrane.

PubMed

Carel, Clément; Marcoux, Julien; Réat, Valérie; Parra, Julien; Latgé, Guillaume; Laval, Françoise; Demange, Pascal; Burlet-Schiltz, Odile; Milon, Alain; Daffé, Mamadou; Tropis, Maryelle G; Renault, Marie A M

2017-04-18

The outer membranes (OMs) of members of the Corynebacteriales bacterial order, also called mycomembranes, harbor mycolic acids and unusual outer membrane proteins (OMPs), including those with α-helical structure. The signals that allow precursors of such proteins to be targeted to the mycomembrane remain uncharacterized. We report here the molecular features responsible for OMP targeting to the mycomembrane of Corynebacterium glutamicum , a nonpathogenic member of the Corynebacteriales order. To better understand the mechanisms by which OMP precursors were sorted in C. glutamicum , we first investigated the partitioning of endogenous and recombinant PorA, PorH, PorB, and PorC between bacterial compartments and showed that they were both imported into the mycomembrane and secreted into the extracellular medium. A detailed investigation of cell extracts and purified proteins by top-down MS, NMR spectroscopy, and site-directed mutagenesis revealed specific and well-conserved posttranslational modifications (PTMs), including O -mycoloylation, pyroglutamylation, and N -formylation, for mycomembrane-associated and -secreted OMPs. PTM site sequence analysis from C. glutamicum OMP and other O -acylated proteins in bacteria and eukaryotes revealed specific patterns. Furthermore, we found that such modifications were essential for targeting to the mycomembrane and sufficient for OMP assembly into mycolic acid-containing lipid bilayers. Collectively, it seems that these PTMs have evolved in the Corynebacteriales order and beyond to guide membrane proteins toward a specific cell compartment.

Identification of specific posttranslational O-mycoloylations mediating protein targeting to the mycomembrane

PubMed Central

Carel, Clément; Réat, Valérie; Parra, Julien; Latgé, Guillaume; Laval, Françoise; Burlet-Schiltz, Odile; Milon, Alain; Daffé, Mamadou; Tropis, Maryelle G.; Renault, Marie A. M.

2017-01-01

The outer membranes (OMs) of members of the Corynebacteriales bacterial order, also called mycomembranes, harbor mycolic acids and unusual outer membrane proteins (OMPs), including those with α-helical structure. The signals that allow precursors of such proteins to be targeted to the mycomembrane remain uncharacterized. We report here the molecular features responsible for OMP targeting to the mycomembrane of Corynebacterium glutamicum, a nonpathogenic member of the Corynebacteriales order. To better understand the mechanisms by which OMP precursors were sorted in C. glutamicum, we first investigated the partitioning of endogenous and recombinant PorA, PorH, PorB, and PorC between bacterial compartments and showed that they were both imported into the mycomembrane and secreted into the extracellular medium. A detailed investigation of cell extracts and purified proteins by top-down MS, NMR spectroscopy, and site-directed mutagenesis revealed specific and well-conserved posttranslational modifications (PTMs), including O-mycoloylation, pyroglutamylation, and N-formylation, for mycomembrane-associated and -secreted OMPs. PTM site sequence analysis from C. glutamicum OMP and other O-acylated proteins in bacteria and eukaryotes revealed specific patterns. Furthermore, we found that such modifications were essential for targeting to the mycomembrane and sufficient for OMP assembly into mycolic acid-containing lipid bilayers. Collectively, it seems that these PTMs have evolved in the Corynebacteriales order and beyond to guide membrane proteins toward a specific cell compartment. PMID:28373551

Analytic energy gradients for orbital-optimized MP3 and MP2.5 with the density-fitting approximation: An efficient implementation.

PubMed

Bozkaya, Uğur

2018-03-15

Efficient implementations of analytic gradients for the orbital-optimized MP3 and MP2.5 and their standard versions with the density-fitting approximation, which are denoted as DF-MP3, DF-MP2.5, DF-OMP3, and DF-OMP2.5, are presented. The DF-MP3, DF-MP2.5, DF-OMP3, and DF-OMP2.5 methods are applied to a set of alkanes and noncovalent interaction complexes to compare the computational cost with the conventional MP3, MP2.5, OMP3, and OMP2.5. Our results demonstrate that density-fitted perturbation theory (DF-MP) methods considered substantially reduce the computational cost compared to conventional MP methods. The efficiency of our DF-MP methods arise from the reduced input/output (I/O) time and the acceleration of gradient related terms, such as computations of particle density and generalized Fock matrices (PDMs and GFM), solution of the Z-vector equation, back-transformations of PDMs and GFM, and evaluation of analytic gradients in the atomic orbital basis. Further, application results show that errors introduced by the DF approach are negligible. Mean absolute errors for bond lengths of a molecular set, with the cc-pCVQZ basis set, is 0.0001-0.0002 Å. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

Purification, characterization and sequence analysis of Omp50,a new porin isolated from Campylobacter jejuni.

PubMed Central

Bolla, J M; Dé, E; Dorez, A; Pagès, J M

2000-01-01

A novel pore-forming protein identified in Campylobacter was purified by ion-exchange chromatography and named Omp50 according to both its molecular mass and its outer membrane localization. We observed a pore-forming ability of Omp50 after re-incorporation into artificial membranes. The protein induced cation-selective channels with major conductance values of 50-60 pS in 1 M NaCl. N-terminal sequencing allowed us to identify the predicted coding sequence Cj1170c from the Campylobacter jejuni genome database as the corresponding gene in the NCTC 11168 genome sequence. The gene, designated omp50, consists of a 1425 bp open reading frame encoding a deduced 453-amino acid protein with a calculated pI of 5.81 and a molecular mass of 51169.2 Da. The protein possessed a 20-amino acid leader sequence. No significant similarity was found between Omp50 and porin protein sequences already determined. Moreover, the protein showed only weak sequence identity with the major outer-membrane protein (MOMP) of Campylobacter, correlating with the absence of antigenic cross-reactivity between these two proteins. Omp50 is expressed in C. jejuni and Campylobacter lari but not in Campylobacter coli. The gene, however, was detected in all three species by PCR. According to its conformation and functional properties, the protein would belong to the family of outer-membrane monomeric porins. PMID:11104668

pH-dependent pore-forming activity of OmpATb from Mycobacterium tuberculosis and characterization of the channel by peptidic dissection.

PubMed

Molle, Virginie; Saint, Nathalie; Campagna, Sylvie; Kremer, Laurent; Lea, Edward; Draper, Philip; Molle, Gérard

2006-08-01

Mycobacteria are characterized by an unusual cell wall that controls nutrient and small hydrophilic compound permeability. Porin-like proteins are necessary to ensure the transport of molecules into the cell. Here, we investigated the pore-forming properties of OmpATb, a porin from Mycobacterium tuberculosis, in lipid bilayers. Multi-channel experiments showed an asymmetric behaviour with channel closures at negative critical voltages (Vc) and a strong decrease in Vc at acidic pH. Single-channel experiments gave conductance values of about 850 +/- 80 pS in 1 M KCl and displayed a weak cationic selectivity in 4-8 pH range. The production and characterization of a series of truncated OmpATb proteins, showed that the central domain (OmpATb73-220) was sufficient to induce the ion channel properties of the native protein in lipid bilayers, i.e. asymmetric insertion, pH-dependent voltage closure, cationic selectivity and similar conductance values in 1 M KCl. Western blot analysis suggests that the presence of OmpATb is only restricted to certain pathogenic species. Therefore, the propensity of channels of native OmpATb to close at low pH may represent an intrinsic property allowing pathogenic mycobacteria to adapt and survive to mildly acidic conditions, such as those encountered within the macrophage phagosome.

What happens with organic micropollutants during UV disinfection in WWTPs? A global perspective from laboratory to full-scale.

PubMed

Paredes, L; Omil, F; Lema, J M; Carballa, M

2018-01-15

The phototransformation of 18 organic micropollutants (OMPs) commonly detected in wastewater treatment plant (WWTP) effluents was examined attempting to explain their fate during UV disinfection in WWTPs. For this purpose, a lab-scale UV reactor (lamp emitting at 254nm) was used to study the influence of the operational conditions (UV dose, temperature and water matrix) on OMPs abatement and disinfection efficiency. Chemical properties of OMPs and the quality of treated effluent were identified as key factors affecting the phototransformation rate of these compounds. Sampling campaigns were carried out at the inlet and outlet of UV systems of three WWTPs, and the results evidenced that only the most photosensitive compounds, such as sulfamethoxazole and diclofenac, are eliminated. Therefore, despite UV treatment is an effective technology to phototransform OMPs, the UV doses typically applied for disinfection (10-50mJ/cm 2 ) are not sufficient to remove them. Consequently, small modifications (increase of UV dose, use of catalysts) should be applied in WWTPs to enhance the abatement of OMPs in UV systems. Copyright © 2017 Elsevier B.V. All rights reserved.

Evanescent wave DNA-aptamer biosensor based on long period gratings for the specific recognition of E. coli outer membrane proteins.

PubMed

Queirós, R B; Gouveia, C; Fernandes, J R A; Jorge, P A S

2014-12-15

An evanescent wave fiber optic sensor for detection of Escherichia coli (E. coli) outer membranes proteins (EcOMPs) using long period gratings (LPGs) as a refractometric platform is presented. The sensing probes were attained by the functionalization of LPGs inscribed in single mode fiber using two different methods of immobilization; electrostatic assembly and covalent binding. The resulting label-free configuration enabled the specific recognition of EcOMPs in water by monitoring the resonance wavelength shift due to refractive index changes induced by binding events. The sensors displayed linear responses in the range of 0.1 nM to 10 nM EcOMPs with sensitivities of -0.1563±0.005 nm decade(-1) [EcOMP, M] (electrostatic method) and -0.1597±0.004 nm decade(-1) [EcOMP, M] (covalent method). The devices could be regenerated (under low pH conditions) with a deviation less than 0.1% for at least three subsequent detection events. The sensors were also applied to spiked environmental water samples. Copyright © 2014 Elsevier B.V. All rights reserved.

DNS of Supersonic Turbulent Flows in a DLR Scramjet Intake

NASA Astrophysics Data System (ADS)

Li, Xinliang; Yu, Changping

2014-11-01

Direct numerical simulation (DNS) of supersonic/hypersonic flow through a DLR scramjet intake GK01 is performed. The free stream Mach numbers are 3, 5 and 7, and the angle of attack is zero degree. The DNS cases are performed by using an optimized MP scheme with adaptive dissipation (OMP-AD) developed by the authors, and the blow-and-suction perturbations near the leading edge are used to trigger the transition. To stabilize the simulation, locally non-linear flittering is used in high-Mach-number case. The transition, separation, and shock-turbulent boundary layer interaction are studied by using both flow visualization and statistical analysis. A new method, OMP-AD, is also addressed in this paper. The OMP-AD scheme is developed by using joint MP method and optimized technique, and the coefficients in the scheme are flexible to show low dissipation in the smoothing region, and to show high robust (but high dissipation) in the large gradient region. Numerical tests show that the OMP-AD is more robust than the original MP schemes, and the numerical dissipation of OMP-AD is very low.

Dissecting the total transition state stabilization provided by amino acid side chains at orotidine 5'-monophosphate decarboxylase: a two-part substrate approach.

PubMed

Barnett, Shonoi A; Amyes, Tina L; Wood, Bryant M; Gerlt, John A; Richard, John P

2008-07-29

Kinetic analysis of decarboxylation catalyzed by S154A, Q215A, and S154A/Q215A mutant yeast orotidine 5'-monophosphate decarboxylases with orotidine 5'-monophosphate (OMP) and with a truncated nucleoside substrate (EO) activated by phosphite dianion shows (1) the side chain of Ser-154 stabilizes the transition state through interactions with the pyrimidine rings of OMP or EO, (2) the side chain of Gln-215 interacts with the phosphodianion group of OMP or with phosphite dianion, and (3) the interloop hydrogen bond between the side chains of Ser-154 and Gln-215 orients the amide side chain of Gln-215 to interact with the phosphodianion group of OMP or with phosphite dianion.

Sustainable rare diseases business and drug access: no time for misconceptions.

PubMed

Rollet, Pierrick; Lemoine, Adrien; Dunoyer, Marc

2013-07-23

Legislative incentives enacted in Europe through the Regulation (EC) No. 141/2000 to incentivize orphan drug development have over the last 12 years constituted a powerful impetus toward R&D directed at the rare diseases population. However, despite therapeutic promises contained in these projects and significant economic impact linked to burgeoning R&D expenditures, the affordability and value of OMPs has become a topic of health policy debate in Europe fueled by the perception that OMPs have high acquisition costs, and by misconceptions around pricing dynamics and rare-diseases business models. In order to maintain sustainable patient access to new and innovative therapies, it is essential to address these misconceptions, and to ensure the successful continuation of a dynamic OMPs R&D within rare-diseases public health policy. Misconceptions abound regarding the pricing of rare diseases drugs and reflect a poor appreciation of the R&D model and the affordability and value of OMPs. Simulation of potential financial returns of small medium sized rare diseases companies focusing on high priced drugs show that their economic returns are likely to be close to their cost of capital. Research in rare diseases is a challenging endeavour characterised by high fixed costs in which companies accrue substantial costs for several years before potentially generating returns from the fruits of their investments. Although heavily dependent upon R&D capabilities of each individual company or R&D organization, continuous flow of R&D financial investment should allow industry to increasingly include efficiencies in research and development in cost considerations to its customers. Industry should also pro-actively work on facilitating development of a specific value based pricing approach to help understanding what constitute value in rare diseases. Policy makers must reward innovation based upon unmet need and patient outcome. Broader understanding by clinicians, the public, and policy makers of the complexity of clinical programs to deliver OMPs to market is required to better comprehend the decisions needed and made by industry. In parallel, an overt effort to consider the impact of public policies on R&D investments is key to enable policy makers to better reconcile the incentives provided by public policy decisions and companies investments decisions in a more positive manner.

Sustainable rare diseases business and drug access: no time for misconceptions

PubMed Central

2013-01-01

Legislative incentives enacted in Europe through the Regulation (EC) No. 141/2000 to incentivize orphan drug development have over the last 12 years constituted a powerful impetus toward R&D directed at the rare diseases population. However, despite therapeutic promises contained in these projects and significant economic impact linked to burgeoning R&D expenditures, the affordability and value of OMPs has become a topic of health policy debate in Europe fueled by the perception that OMPs have high acquisition costs, and by misconceptions around pricing dynamics and rare-diseases business models. In order to maintain sustainable patient access to new and innovative therapies, it is essential to address these misconceptions, and to ensure the successful continuation of a dynamic OMPs R&D within rare-diseases public health policy. Misconceptions abound regarding the pricing of rare diseases drugs and reflect a poor appreciation of the R&D model and the affordability and value of OMPs. Simulation of potential financial returns of small medium sized rare diseases companies focusing on high priced drugs show that their economic returns are likely to be close to their cost of capital. Research in rare diseases is a challenging endeavour characterised by high fixed costs in which companies accrue substantial costs for several years before potentially generating returns from the fruits of their investments. Although heavily dependent upon R&D capabilities of each individual company or R&D organization, continuous flow of R&D financial investment should allow industry to increasingly include efficiencies in research and development in cost considerations to its customers. Industry should also pro-actively work on facilitating development of a specific value based pricing approach to help understanding what constitute value in rare diseases. Policy makers must reward innovation based upon unmet need and patient outcome. Broader understanding by clinicians, the public, and policy makers of the complexity of clinical programs to deliver OMPs to market is required to better comprehend the decisions needed and made by industry. In parallel, an overt effort to consider the impact of public policies on R&D investments is key to enable policy makers to better reconcile the incentives provided by public policy decisions and companies investments decisions in a more positive manner. PMID:23879976

Continuation of long-term global SO2 pollution monitoring from OMI to OMPS

NASA Astrophysics Data System (ADS)

Zhang, Yan; Li, Can; Krotkov, Nickolay A.; Joiner, Joanna; Fioletov, Vitali; McLinden, Chris

2017-04-01

Over the past 20 years, advances in satellite remote sensing of pollution-relevant species have made space-borne observations an increasingly important part of atmospheric chemistry research and air quality management. This progress has been facilitated by advanced UV-vis spectrometers, such as the Ozone Monitoring Instrument (OMI) on board the NASA Earth Observing System (EOS) Aura satellite, and continues with new instruments, such as the Ozone Mapping and Profiler Suite (OMPS) on board the NASA-NOAA Suomi National Polar-orbiting Partnership (SNPP) satellite. In this study, we demonstrate that it is possible, using our state-of-the-art principal component analysis (PCA) retrieval technique, to continue the long-term global SO2 pollution monitoring started by OMI with the current and future OMPS instruments that will fly on the NOAA Joint Polar Satellite System (JPSS) 1, 2, 3, and 4 satellites in addition to SNPP, with a very good consistency of retrievals from these instruments. Since OMI SO2 data have been primarily used for (1) providing regional context on air pollution and long-range transport on a daily basis and (2) providing information on point emission sources on an annual basis after data averaging, we focused on these two aspects in our OMI-OMPS comparisons. Four years of retrievals (2012-2015) have been compared for three regions: eastern China, Mexico, and South Africa. In general, the comparisons show relatively high correlations (r = 0. 79-0.96) of daily regional averaged SO2 mass between the two instruments and near-unity regression slopes (0.76-0.97). The annual averaged SO2 loading differences between OMI and OMPS are small (< 0.03 Dobson unit (DU) over South Africa and up to 0.1 DU over eastern China). We also found a very good correlation (r = 0. 92-0.97) in the spatial distribution of annual averaged SO2 between OMI and OMPS over the three regions during 2012-2015. The emissions from ˜ 400 SO2 sources calculated with the two instruments also show a very good correlation (r = ˜ 0.9) in each year during 2012-2015. OMPS-detected SO2 point source emissions are slightly lower than those from OMI, but OMI-OMPS differences decrease with increasing strength of source. The OMI-OMPS SO2 mass differences on a pixel by pixel (daily) basis in each region can show substantial differences. The two instruments have a spatial correlation coefficient of 0.7 or better on < ˜ 50 % of the days. It is worth noting that consistent SO2 retrievals were achieved without any explicit adjustments to OMI or OMPS radiance data and that the retrieval agreement may be further improved by introducing a more comprehensive Jacobian lookup table than is currently used.

Ozone Mapping and Profiler Suite: using mission performance data to refine predictive contamination modeling

NASA Astrophysics Data System (ADS)

Devaud, Genevieve; Jaross, Glen

2014-09-01

On October 28, 2011, the Suomi National Polar-orbiting Partnership (Suomi NPP) satellite launched at Vandenberg Air Force base aboard a United Launch Alliance Delta II rocket. Included among the five instruments was the Ozone Mapping and Profiler Suite (OMPS), an advanced suite of three hyperspectral instruments built by Ball Aerospace and Technologies Corporation (BATC) for the NASA Goddard Space Flight Center. Molecular transport modeling is used to predict optical throughput changes due to contaminant accumulation to ensure performance margin to End Of Life. The OMPS Nadir Profiler, operating at the lowest wavelengths of 250 - 310 nm, is most sensitive to contaminant accumulation. Geometry, thermal profile and material properties must be accurately modeled in order to have confidence in the results, yet it is well known that the complex chemistry and process dependent variability of aerospace materials presents a substantial challenge to the modeler. Assumptions about the absorption coefficients, desorption and diffusion kinetics of outgassing species from polymeric materials dramatically affect the model predictions, yet it is rare indeed that on-mission data is analyzed at a later date as a means to compare with modeling results. Optical throughput measurements for the Ozone and Mapping Profiler Suite on the Suomi NPP Satellite indicate that optical throughput degradation between day 145 and day 858 is less than 0.5%. We will show how assumptions about outgassing rates and desorption energies, in particular, dramatically affect the modeled optical throughput and what assumptions represent the on-orbit data.

Comparative study of proteasome inhibitory, synergistic antibacterial, synergistic anticandidal, and antioxidant activities of gold nanoparticles biosynthesized using fruit waste materials.

PubMed

Patra, Jayanta Kumar; Baek, Kwang-Hyun

The aim of this study was to compare the biological synthesis of gold nanoparticles (AuNPs) generated using the aqueous extracts of outer oriental melon peel (OMP) and peach. The synthesized OMP-AuNPs and peach extract (PE)-AuNPs were characterized by ultraviolet-visible spectroscopy, field emission scanning electron microscopy, energy dispersive X-ray analysis, X-ray powder diffraction, Fourier transform infrared spectroscopy, and thermogravimetric analysis. The surface plasmon resonance spectra were obtained at 545 nm and 540 nm for OMP-AuNPs and PE-AuNPs, respectively. The estimated absolute crystallite size of the synthesized AuNPs was calculated to be 78.11 nm for OMP-AuNPs and 39.90 nm for PE-AuNPs based on the Scherer equation of the X-ray powder diffraction peaks. Fourier transform infrared spectroscopy results revealed the involvement of bioactive compounds present in OMP and peach extracts in the synthesis and stabilization of synthesized AuNPs. Both the OMP-AuNPs and PE-AuNPs showed a strong antibacterial synergistic activity when combined with kanamycin (9.38-20.45 mm inhibition zones) and rifampicin (9.52-25.23 mm inhibition zones), and they also exerted a strong synergistic anticandidal activity (10.09-15.47 mm inhibition zones) when combined with amphotericin B against five pathogenic Candida species. Both the OMP-AuNPs and PE-AuNPs exhibited a strong antioxidant potential in terms of 1,1-diphenyl-2-picrylhydraxyl radical scavenging, nitric oxide scavenging, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) radical scavenging, and a reducing power, along with a strong proteasome inhibitory potential that could be useful in cancer drug delivery and cancer treatments. The PE-AuNPs showed comparatively higher activity than OMP-AuNPs, which could be attributed to the presence of rich bioactive compounds in the PE that acted as reducing and capping agents in the synthesis of PE-AuNPs. Overall, the results of the current investigation highlighted a novel green technology for the synthesis of AuNPs using food waste materials and their potential applications in the biomedical, pharmaceutical, and cosmetic industries.

Detoxified Endotoxin Vaccine (J5dLPS/OMP) Protects Mice Against Lethal Respiratory Challenge with Francisella tularensis SchuS4

PubMed Central

Gregory, Stephen H.; Chen, Wilbur H.; Mott, Stephanie; Palardy, John E.; Parejo, Nicholas A.; Heninger, Sara; Anderson, Christine A.; Artenstein, Andrew W.; Opal, Steven M.; Cross, Alan S.

2010-01-01

Francisella tularensis is a category A select agent. J5dLPS/OMP is a novel vaccine construct consisting of detoxified, O-polysaccharide side chain-deficient, lipopolysaccharide non-covalently complexed with the outer membrane protein of N. meningitidis group B. Immunization elicits hightiter polyclonal antibodies specific for the highly-conserved epitopes expressed within the glycolipid core that constitutes gram-negative bacteria (e.g., F. tularensis). Mice immunized intranasally with J5dLPS/OMP exhibited protective immunity to intratracheal challenge with the live vaccine strain, as well as the highly-virulent SchuS4 strain, of F. tularensis. The efficacy of J5dLPS/OMP vaccine suggests its potential utility in immunizing the general population against several different gram-negative select agents concurrently. PMID:20170768

Novel utilization of the outer membrane proteins for the identification and differentiation of pathogenic versus nonpathogenic microbial strains using mass spectrometry-based proteomics approach

NASA Astrophysics Data System (ADS)

Jabbour, Rabih E.; Wade, Mary; Deshpande, Samir V.; McCubbin, Patrick; Snyder, A. Peter; Bevilacqua, Vicky

2012-06-01

Mass spectrometry based proteomic approaches are showing promising capabilities in addressing various biological and biochemical issues. Outer membrane proteins (OMPs) are often associated with virulence in gram-negative pathogens and could prove to be excellent model biomarkers for strain level differentiation among bacteria. Whole cells and OMP extracts were isolated from pathogenic and non-pathogenic strains of Francisella tularensis, Burkholderia thailandensis, and Burkholderia mallei. OMP extracts were compared for their ability to differentiate and delineate the correct database organism to an experimental sample and for the degree of dissimilarity to the nearest-neighbor database strains. This study addresses the comparative experimental proteome analyses of OMPs vs. whole cell lysates on the strain-level discrimination among gram negative pathogenic and non-pathogenic strains.

Chloroplast outer envelope protein P39 in Arabidopsis thaliana belongs to the Omp85 protein family.

PubMed

Hsueh, Yi-Ching; Flinner, Nadine; Gross, Lucia E; Haarmann, Raimund; Mirus, Oliver; Sommer, Maik S; Schleiff, Enrico

2017-08-01

Proteins of the Omp85 family chaperone the membrane insertion of β-barrel-shaped outer membrane proteins in bacteria, mitochondria, and probably chloroplasts and facilitate the transfer of nuclear-encoded cytosolically synthesized preproteins across the outer envelope of chloroplasts. This protein family is characterized by N-terminal polypeptide transport-associated (POTRA) domains and a C-terminal membrane-embedded β-barrel. We have investigated a recently identified Omp85 family member of Arabidopsis thaliana annotated as P39. We show by in vitro and in vivo experiments that P39 is localized in chloroplasts. The electrophysiological properties of P39 are consistent with those of other Omp85 family members confirming the sequence based assignment of P39 to this family. Bioinformatic analysis showed that P39 lacks any POTRA domain, while a complete 16 stranded β-barrel including the highly conserved L6 loop is proposed. The electrophysiological properties are most comparable to Toc75-V, which is consistent with the phylogenetic clustering of P39 in the Toc75-V rather than the Toc75-III branch of the Omp85 family tree. Taken together P39 forms a pore with Omp85 family protein characteristics. The bioinformatic comparison of the pore region of Toc75-III, Toc75-V, and P39 shows distinctions of the barrel region most likely related to function. Proteins 2017; 85:1391-1401. © 2014 Wiley Periodicals, Inc. © 2014 Wiley Periodicals, Inc.

Refolding of the recombinant protein OmpA70 from Leptospira interrogans from inclusion bodies using high hydrostatic pressure and partial characterization of its immunological properties.

PubMed

Fraga, Tatiana R; Chura-Chambi, Rosa M; Gonçales, Amane P; Morais, Zenaide M; Vasconcellos, Sílvio A; Morganti, Ligia; Martins, Elizabeth A L

2010-07-20

Leptospira is the etiological agent of leptospirosis, a life-threatening disease that affects human populations worldwide. Available vaccines have demonstrated limited effectiveness, and therapeutic interventions are complicated by the difficulty of establishing an early diagnosis. The genome of Leptospira strains was sequenced, and bioinformatic analyses revealed potential vaccine and serodiagnosis candidates. The present work studied OmpA70, a putative outer membrane protein from Leptospira interrogans serovar Copenhageni that combines structural features of Loa22, the first genetically defined virulence factor in Leptospira, and Lp49, a protein that reacts with sera from early and convalescent patients. Recombinant OmpA was produced in Escherichia coli in an insoluble form. Considering the importance of the structural integrity of a protein to confer immune protection, high hydrostatic pressure (HHP) was used to refold OmpA70 aggregated as inclusion bodies. HHP was applied in association with redox-shuffling reagents (oxidized and reduced glutathione) and guanidine hydrochloride or l-arginine. About 40% of the protein was refolded by applying 200MPa for 16h in concentrations of l-arginine above 0.4M. Circular dichroism revealed the presence of secondary structure. OmpA70 has immunogenic and antigenic properties as high antibody titers were seen after immunization with this protein, and sera from infected hamsters reacted with soluble OmpA70. Copyright (c) 2010 Elsevier B.V. All rights reserved.

Differences in outer membrane proteins of the lymphogranuloma venereum and trachoma biovars of Chlamydia trachomatis

DOE Office of Scientific and Technical Information (OSTI.GOV)

Batteiger, B.E.; Jones, R.B.

1985-11-01

The lymphogranuloma venereum (LGV) and trachoma biovars of Chlamydia trachomatis exhibit differences in biological properties both in vivo and in vitro. To identify analogous biochemical differences, the authors studied the molecular charges of chlamydial outer membrane proteins (OMPs) by means of isoelectric focusing and nonequilibrium pH gradient electrophoresis. Analysis of proteins of whole elementary bodies biosynthetically labeled with L-(35S)cysteine revealed that most chlamydial proteins were neutral or acidic. The major OMPs (MOMPs) of all strains tested were acidic and had apparent isoelectric points (pIs) that varied within narrow limits despite differences in molecular mass of up to 3,000 daltons (Da).more » However, a low-molecular-mass cysteine-rich OMP analogous to that previously described for Chlamydia psittaci varied consistently in molecular mass (12,500 versus 12,000 Da) and pI (5.4 versus 6.9) between LGV strains and trachoma strains, respectively. OMPs with a molecular mass of 60,000 Da in the trachoma biovar strains had pIs in the 7.3 to 7.7 range. However, analogous OMPs in the LGV strains existed as a doublet with a molecular mass of about 60,000 Da. These data indicate substantial differences in biochemical characteristics of analogous OMPs in the LGV and trachoma biovars. Such differences are the first structural differences described between LGV and trachoma strains which support their distinction into separate biovars and may be related to some of their biological differences.« less

Chloroform-Methanol Residue of Coxiella burnetii Markedly Potentiated the Specific Immunoprotection Elicited by a Recombinant Protein Fragment rOmpB-4 Derived from Outer Membrane Protein B of Rickettsia rickettsii in C3H/HeN Mice

PubMed Central

Gong, Wenping; Wang, Pengcheng; Xiong, Xiaolu; Jiao, Jun; Yang, Xiaomei; Wen, Bohai

2015-01-01

The obligate intracellular bacteria, Rickettsia rickettsii and Coxiella burnetii, are the potential agents of bio-warfare/bio-terrorism. Here C3H/HeN mice were immunized with a recombinant protein fragment rOmp-4 derived from outer membrane protein B, a major protective antigen of R. rickettsii, combined with chloroform-methanol residue (CMR) extracted from phase I C. burnetii organisms, a safer Q fever vaccine. These immunized mice had significantly higher levels of IgG1 and IgG2a to rOmpB-4 and interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α), two crucial cytokines in resisting intracellular bacterial infection, as well as significantly lower rickettsial loads and slighter pathological lesions in organs after challenge with R. rickettsii, compared with mice immunized with rOmpB-4 or CMR alone. Additionally, after challenge with C. burnetii, the coxiella loads in the organs of these mice were significantly lower than those of mice immunized with rOmpB-4 alone. Our results prove that CMR could markedly potentiate enhance the rOmpB-4-specific immunoprotection by promoting specific and non-specific immunoresponses and the immunization with the protective antigen of R. rickettsii combined with CMR of C. burnetii could confer effective protection against infection of R. rickettsii or C. burnetii. PMID:25909586

Tomographic retrievals of ozone with the OMPS Limb Profiler: algorithm description and preliminary results

NASA Astrophysics Data System (ADS)

Zawada, Daniel J.; Rieger, Landon A.; Bourassa, Adam E.; Degenstein, Douglas A.

2018-04-01

Measurements of limb-scattered sunlight from the Ozone Mapping and Profiler Suite Limb Profiler (OMPS-LP) can be used to obtain vertical profiles of ozone in the stratosphere. In this paper we describe a two-dimensional, or tomographic, retrieval algorithm for OMPS-LP where variations are retrieved simultaneously in altitude and the along-orbital-track dimension. The algorithm has been applied to measurements from the center slit for the full OMPS-LP mission to create the publicly available University of Saskatchewan (USask) OMPS-LP 2D v1.0.2 dataset. Tropical ozone anomalies are compared with measurements from the Microwave Limb Sounder (MLS), where differences are less than 5 % of the mean ozone value for the majority of the stratosphere. Examples of near-coincident measurements with MLS are also shown, and agreement at the 5 % level is observed for the majority of the stratosphere. Both simulated retrievals and coincident comparisons with MLS are shown at the edge of the polar vortex, comparing the results to a traditional one-dimensional retrieval. The one-dimensional retrieval is shown to consistently overestimate the amount of ozone in areas of large horizontal gradients relative to both MLS and the two-dimensional retrieval.

Co-regulation of polysaccharide production, motility, and expression of type III secretion genes by EnvZ/OmpR and GrrS/GrrA systems in Erwinia amylovora.

PubMed

Li, Wenting; Ancona, Veronica; Zhao, Youfu

2014-02-01

The EnvZ/OmpR and GrrS/GrrA systems, two widely distributed two-component systems in gamma-Proteobacteria, negatively control amylovoran biosynthesis in Erwinia amylovora, and the two systems regulate motility in an opposing manner. In this study, we examined the interplay of EnvZ/OmpR and GrrS/GrrA systems in controlling various virulence traits in E. amylovora. Results showed that amylovoran production was significantly higher when both systems were inactivated, indicating that the two systems act as negative regulators and their combined effect on amylovoran production appears to be enhanced. In contrast, reduced motility was observed when both systems were deleted as compared to that of grrA/grrS mutants and WT strain, indicating that the two systems antagonistically regulate motility in E. amylovora. In addition, glycogen accumulation was much higher in envZ/ompR and two triple mutants than that of grrS/grrA mutants and WT strain, suggesting that EnvZ/OmpR plays a dominant role in regulating glycogen accumulation, whereas levan production was significantly lower in the grrS/grrA and two triple mutants as compared with that of WT and envZ/ompR mutants, indicating that GrrS/GrrA system dominantly controls levan production. Furthermore, both systems negatively regulated expression of three type III secretion (T3SS) genes and their combined negative effect on hrp-T3SS gene expression increased when both systems were deleted. These results demonstrated that EnvZ/OmpR and GrrS/GrrA systems co-regulate various virulence factors in E. amylovora by still unknown mechanisms or through different target genes, sRNAs, or proteins, indicating that a complex regulatory network may be involved, which needs to be further explored.

The TOMS V9 Algorithm for OMPS Nadir Mapper Total Ozone: An Enhanced Design That Ensures Data Continuity

NASA Astrophysics Data System (ADS)

Haffner, D. P.; McPeters, R. D.; Bhartia, P. K.; Labow, G. J.

2015-12-01

The TOMS V9 total ozone algorithm will be applied to the OMPS Nadir Mapper instrument to supersede the exisiting V8.6 data product in operational processing and re-processing for public release. Becuase the quality of the V8.6 data is already quite high, enchancements in V9 are mainly with information provided by the retrieval and simplifcations to the algorithm. The design of the V9 algorithm has been influenced by improvements both in our knowledge of atmospheric effects, such as those of clouds made possible by studies with OMI, and also limitations in the V8 algorithms applied to both OMI and OMPS. But the namesake instruments of the TOMS algorithm are substantially more limited in their spectral and noise characterisitics, and a requirement of our algorithm is to also apply the algorithm to these discrete band spectrometers which date back to 1978. To achieve continuity for all these instruments, the TOMS V9 algorithm continues to use radiances in discrete bands, but now uses Rodgers optimal estimation to retrieve a coarse profile and provide uncertainties for each retrieval. The algorithm remains capable of achieving high accuracy results with a small number of discrete wavelengths, and in extreme cases, such as unusual profile shapes and high solar zenith angles, the quality of the retrievals is improved. Despite the intended design to use limited wavlenegths, the algorithm can also utilitze additional wavelengths from hyperspectral sensors like OMPS to augment the retreival's error detection and information content; for example SO2 detection and correction of Ring effect on atmospheric radiances. We discuss these and other aspects of the V9 algorithm as it will be applied to OMPS, and will mention potential improvements which aim to take advantage of a synergy with OMPS Limb Profiler and Nadir Mapper to further improve the quality of total ozone from the OMPS instrument.

Development and Comparative Evaluation of a Plate Enzyme-Linked Immunosorbent Assay Based on Recombinant Outer Membrane Antigens Omp28 and Omp31 for Diagnosis of Human Brucellosis

PubMed Central

Tiwari, Sapana; Kumar, Ashu; Mangalgi, Smita; Rathod, Vedika; Prakash, Archana; Barua, Anita; Arora, Sonia; Sathyaseelan, Kannusamy

2013-01-01

Brucellosis is an important zoonotic infectious disease of humans and livestock with worldwide distribution and is caused by bacteria of the genus Brucella. The diagnosis of brucellosis always requires laboratory confirmation by either isolation of pathogens or detection of specific antibodies. The conventional serological tests available for the diagnosis of brucellosis are less specific and show cross-reactivity with other closely related organisms. These tests also necessitate the handling of Brucella species for antigen preparation. Therefore, there is a need to develop reliable, rapid, and user-friendly systems for disease diagnosis and alternatives to vaccine approaches. Keeping in mind the importance of brucellosis as an emerging infection and the prevalence in India, we carried out the present study to compare the recombinant antigens with the native antigens (cell envelope and sonicated antigen) of Brucella for diagnosis of human brucellosis by an indirect plate enzyme-linked immunosorbent assay (ELISA). Recombinant outer membrane protein 28 (rOmp28) and rOmp31 antigens were cloned, expressed, and purified in the bacterial expression system, and the purified proteins were used as antigens. Indirect plate ELISAs were then performed and standardized for comparison of the reactivities of recombinant and native antigens against the 433 clinical samples submitted for brucellosis testing, 15 culture-positive samples, and 20 healthy donor samples. The samples were separated into four groups based on their positivity to rose bengal plate agglutination tests (RBPTs), standard tube agglutination tests (STATs), and 2-mercaptoethanol (2ME) tests. The sensitivities and specificities of all the antigens were calculated, and the rOmp28 antigen was found to be more suitable for the clinical diagnosis of brucellosis than the rOmp31 antigen and native antigens. The rOmp28-based ELISA showed a very high degree of agreement with the conventional agglutination tests and promising results for further use in clinical screening and serodiagnosis of human brucellosis. PMID:23761658

Immunogenicity in chickens with orally administered recombinant chicken-borne Lactobacillus saerimneri expressing FimA and OmpC antigen of O78 avian pathogenic Escherichia coli.

PubMed

Ma, Sun-Ting; Ding, Guo-Jie; Huang, Xue-Wei; Wang, Zi-Wei; Wang, Li; Yu, Mei-Ling; Shi, Wen; Jiang, Yan-Ping; Tang, Li-Jie; Xu, Yi-Gang; Li, Yi-Jing

2018-03-01

Avian colibacillosis is responsible for economic losses to poultry producers worldwide. To combat this, we aimed to develop an effective oral vaccine for chicken against O78 avian pathogenic Escherichia coli (APEC) infection through a Lactobacillus delivery system. Eight Lactobacillus strains isolated from the intestines of broiler chickens were evaluated based on their in vitro adherence ability to assess their potential as a delivery vector. Fimbrial subunit A (FimA) and outer-membrane protein C (OmpC) of APEC with and without fusion to dendritic cell-targeting peptide (DCpep) and microfold cell-targeting peptide (Co1) were displayed on the surface of Lactobacillus saerimneri M-11 and yielded vaccine groups (pPG-ompC-fimA/M-11 and pPG-ompC-fimA-Co1-DCpep/M-11, respectively). The colonization of the recombinant strains in vivo was assessed and the immunogenicity and protective efficacy of orally administered recombinant strains in chickens were evaluated. The colonization of the recombinant strains in vivo revealed no significant differences between the recombinant and wild-type strains. Chickens orally administered with vaccine groups showed significantly higher levels of OmpC/FimA-specific IgG in serum and mucosal IgA in cecum lavage, nasal lavage and stool compared to the pPG/M-11 group. After challenge with APEC CVCC1553, better protective efficacy was observed in chickens orally immunized with pPG-ompC-fimA/M-11 and pPG-ompC-fimA-Co1-DCpep/M-11, but no significant differences were observed between the two groups. Recombinant chicken-borne L. saerimneri M-11 showed good immunogenicity in chickens, suggesting that it may be a promising vaccine candidate against APEC infections. However, the activity of mammalian DCpep and Co1 was not significant in chickens.

Effect of Porins and blaKPC Expression on Activity of Imipenem with Relebactam in Klebsiella pneumoniae: Can Antibiotic Combinations Overcome Resistance?

PubMed

Balabanian, Gregory; Rose, Michael; Manning, Nyla; Landman, David; Quale, John

2018-05-21

Imipenem with relebactam is a novel β-lactam-β-lactamase inhibitor that has activity against most KPC-producing Enterobacteriaceae. Using 10 isolates of KPC-possessing Klebsiella pneumoniae, we assessed the relationship between imipenem-relebactam minimum inhibitory concentrations (MICs) and mechanisms known to contribute to antimicrobial resistance. The effect of adding a second agent was assessed by time-kill experiments. Mutations affecting the genes encoding porins ompK35 and ompK36 and identification of β-lactamases were assessed by PCR. Expression of bla KPC and acrB was assessed by real-time reverse-transcriptase (RT)-PCR, and production of OmpK36 by SDS-PAGE. Time-kill studies were performed using combinations of imipenem-relebactam with polymyxin B, amikacin, or tigecycline. Seven isolates having a disruption in a single porin, or in neither porin, remained susceptible to imipenem-relebactam. The addition of a second agent did not improve the activity of imipenem-relebactam for these isolates, although the addition of tigecycline was antagonistic for three isolates. Three isolates had major disruptions in both ompK35 and ompK36 that correlated with reduced activity of imipenem-relebactam (MICs 2/4, 8/4, and 512/4 μg/mL). Two of these isolates also had overexpression of bla KPC , including the isolate with the highest MIC. These isolates were also resistant to polymyxin B and amikacin. The addition of amikacin provided both synergistic and bactericidal activity for the two more resistant isolates. The activity of imipenem-relebactam against K. pneumoniae is affected by major disruptions of both ompK35 and ompK36 and by expression of the KPC gene. Combining imipenem-relebactam with an aminoglycoside may be a promising approach for isolates with reduced susceptibility to imipenem-relebactam.

Escherichia coli K1 inhibits proinflammatory cytokine induction in monocytes by preventing NF-kappaB activation.

PubMed

Selvaraj, Suresh K; Prasadarao, Nemani V

2005-08-01

Phagocytes are well-known effectors of the innate immune system to produce proinflammatory cytokines and chemokines such as tumor necrosis factor alpha (TNF-alpha), interleukin (IL)-1beta, and IL-8 during infections. Here, we show that infection of monocytes with wild-type Escherichia coli K1, which causes meningitis in neonates, suppresses the production of cytokines and chemokines (TNF-alpha, regulated on activation, normal T expressed and secreted, macrophage-inflammatory protein-1beta, IL-1beta, and IL-8). In contrast, infection of monocytes with a mutant E. coli, which lacks outer membrane protein A (OmpA- E. coli) resulted in robust production of cytokines and chemokines. Wild-type E. coli K1 (OmpA+ E. coli) prevented the phosphorylation and its degradation of inhibitor of kappaB, thereby blocking the translocation of nuclear factor (NF)-kappaB to the nucleus. OmpA+ E. coli-infected cells, subsequently subjected to lipopolysaccharide challenge, were crippled severely in their ability to activate NF-kappaB to induce cytokine/chemokine production. Selective inhibitors of the extracellular signal-regulated kinase (ERK) 1/2 pathway and p38 mitogen-activated protein kinase (MAPK), but not Jun N-terminal kinase, significantly reduced the activation of NF-kappaB and the production of cytokines and chemokines induced by OmpA- E. coli, indicating a role for these kinases in the NF-kappaB/cytokine pathway. It is interesting that the phosphorylation of ERK 1/2 and p38 MAPK was notably reduced in monocytes infected with OmpA+ E. coli when compared with monocytes infected with OmpA- E. coli, suggesting that the modulation of upstream events common for NF-kappaB and MAPKs by the bacterium is possible. The ability of OmpA+ E. coli K1 to inhibit the macrophage response temporarily may enable bacterial survival and growth within the host for the onset of meningitis by E. coli K1.

Methylation and in vivo expression of the surface-exposed Leptospira interrogans outer-membrane protein OmpL32.

PubMed

Eshghi, Azad; Pinne, Marija; Haake, David A; Zuerner, Richard L; Frank, Ami; Cameron, Caroline E

2012-03-01

Recent studies have revealed that bacterial protein methylation is a widespread post-translational modification that is required for virulence in selected pathogenic bacteria. In particular, altered methylation of outer-membrane proteins has been shown to modulate the effectiveness of the host immune response. In this study, 2D gel electrophoresis combined with MALDI-TOF MS identified a Leptospira interrogans serovar Copenhageni strain Fiocruz L1-130 protein, corresponding to ORF LIC11848, which undergoes extensive and differential methylation of glutamic acid residues. Immunofluorescence microscopy implicated LIC11848 as a surface-exposed outer-membrane protein, prompting the designation OmpL32. Indirect immunofluorescence microscopy of golden Syrian hamster liver and kidney sections revealed expression of OmpL32 during colonization of these organs. Identification of methylated surface-exposed outer-membrane proteins, such as OmpL32, provides a foundation for delineating the role of this post-translational modification in leptospiral virulence.

BamA β16C strand and periplasmic turns are critical for outer membrane protein insertion and assembly.

PubMed

Gu, Yinghong; Zeng, Yi; Wang, Zhongshan; Dong, Changjiang

2017-11-21

Outer membrane (OM) β-barrel proteins play important roles in importing nutrients, exporting wastes and conducting signals in Gram-negative bacteria, mitochondria and chloroplasts. The outer membrane proteins (OMPs) are inserted and assembled into the OM by OMP85 family proteins. In Escherichia coli , the β-barrel assembly machinery (BAM) contains four lipoproteins such as BamB, BamC, BamD and BamE, and one OMP BamA, forming a 'top hat'-like structure. Structural and functional studies of the E. coli BAM machinery have revealed that the rotation of periplasmic ring may trigger the barrel β1C-β6C scissor-like movement that promote the unfolded OMP insertion without using ATP. Here, we report the BamA C-terminal barrel structure of Salmonella enterica Typhimurium str. LT2 and functional assays, which reveal that the BamA's C-terminal residue Trp, the β16C strand of the barrel and the periplasmic turns are critical for the functionality of BamA. These findings indicate that the unique β16C strand and the periplasmic turns of BamA are important for the outer membrane insertion and assembly. The periplasmic turns might mediate the rotation of the periplasmic ring to the scissor-like movement of BamA β1C-β6C, triggering the OMP insertion. These results are important for understanding the OMP insertion in Gram-negative bacteria, as well as in mitochondria and chloroplasts. © 2017 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Interaction of Zwitterionic Penicillins with the OmpF Channel Facilitates Their Translocation

PubMed Central

Danelon, Christophe; Nestorovich, Ekaterina M.; Winterhalter, Mathias; Ceccarelli, Matteo; Bezrukov, Sergey M.

2006-01-01

To study translocation of β-lactam antibiotics of different size and charge across the outer bacterial membrane, we combine an analysis of ion currents through single trimeric outer membrane protein F (OmpF) porins in planar lipid bilayers with molecular dynamics simulations. Because the size of penicillin molecules is close to the size of the narrowest part of the OmpF pore, penicillins occlude the pore during their translocation. Favorably interacting penicillins cause time-resolvable transient blockages of the small-ion current through the channel and thereby provide information about their dynamics within the pore. Analyzing these random fluctuations, we find that ampicillin and amoxicillin have a relatively high affinity for OmpF. In contrast, no or only a weak interaction is detected for carbenicillin, azlocillin, and piperacillin. Molecular dynamics simulations suggest a possible pathway of these drugs through the OmpF channel and rationalize our experimental findings. For zwitterionic ampicillin and amoxicillin, we identify a region of binding sites near the narrowest part of the channel pore. Interactions with these sites partially compensate for the entropic cost of drug confinement by the channel. Whereas azlocillin and piperacillin are clearly too big to pass through the channel constriction, dianionic carbenicillin does not find an efficient binding region in the constriction zone. Carbenicillin's favorable interactions are limited to the extracellular vestibule. These observations confirm our earlier suggestion that a set of high-affinity sites at the narrowest part of the OmpF channel improves a drug's ability to cross the membrane via the pore. PMID:16339889

Novel molecular markers of Chlamydia pecorum genetic diversity in the koala (Phascolarctos cinereus)

PubMed Central

2011-01-01

Background Chlamydia pecorum is an obligate intracellular bacterium and the causative agent of reproductive and ocular disease in several animal hosts including koalas, sheep, cattle and goats. C. pecorum strains detected in koalas are genetically diverse, raising interesting questions about the origin and transmission of this species within koala hosts. While the ompA gene remains the most widely-used target in C. pecorum typing studies, it is generally recognised that surface protein encoding genes are not suited for phylogenetic analysis and it is becoming increasingly apparent that the ompA gene locus is not congruent with the phylogeny of the C. pecorum genome. Using the recently sequenced C. pecorum genome sequence (E58), we analysed 10 genes, including ompA, to evaluate the use of ompA as a molecular marker in the study of koala C. pecorum genetic diversity. Results Three genes (incA, ORF663, tarP) were found to contain sufficient nucleotide diversity and discriminatory power for detailed analysis and were used, with ompA, to genotype 24 C. pecorum PCR-positive koala samples from four populations. The most robust representation of the phylogeny of these samples was achieved through concatenation of all four gene sequences, enabling the recreation of a "true" phylogenetic signal. OmpA and incA were of limited value as fine-detailed genetic markers as they were unable to confer accurate phylogenetic distinctions between samples. On the other hand, the tarP and ORF663 genes were identified as useful "neutral" and "contingency" markers respectively, to represent the broad evolutionary history and intra-species genetic diversity of koala C. pecorum. Furthermore, the concatenation of ompA, incA and ORF663 sequences highlighted the monophyletic nature of koala C. pecorum infections by demonstrating a single evolutionary trajectory for koala hosts that is distinct from that seen in non-koala hosts. Conclusions While the continued use of ompA as a fine-detailed molecular marker for epidemiological analysis appears justified, the tarP and ORF663 genes also appear to be valuable markers of phylogenetic or biogeographic divisions at the C. pecorum intra-species level. This research has significant implications for future typing studies to understand the phylogeny, genetic diversity, and epidemiology of C. pecorum infections in the koala and other animal species. PMID:21496349

Comparative study of proteasome inhibitory, synergistic antibacterial, synergistic anticandidal, and antioxidant activities of gold nanoparticles biosynthesized using fruit waste materials

PubMed Central

Patra, Jayanta Kumar; Baek, Kwang-Hyun

2016-01-01

The aim of this study was to compare the biological synthesis of gold nanoparticles (AuNPs) generated using the aqueous extracts of outer oriental melon peel (OMP) and peach. The synthesized OMP-AuNPs and peach extract (PE)-AuNPs were characterized by ultraviolet–visible spectroscopy, field emission scanning electron microscopy, energy dispersive X-ray analysis, X-ray powder diffraction, Fourier transform infrared spectroscopy, and thermogravimetric analysis. The surface plasmon resonance spectra were obtained at 545 nm and 540 nm for OMP-AuNPs and PE-AuNPs, respectively. The estimated absolute crystallite size of the synthesized AuNPs was calculated to be 78.11 nm for OMP-AuNPs and 39.90 nm for PE-AuNPs based on the Scherer equation of the X-ray powder diffraction peaks. Fourier transform infrared spectroscopy results revealed the involvement of bioactive compounds present in OMP and peach extracts in the synthesis and stabilization of synthesized AuNPs. Both the OMP-AuNPs and PE-AuNPs showed a strong antibacterial synergistic activity when combined with kanamycin (9.38–20.45 mm inhibition zones) and rifampicin (9.52–25.23 mm inhibition zones), and they also exerted a strong synergistic anticandidal activity (10.09–15.47 mm inhibition zones) when combined with amphotericin B against five pathogenic Candida species. Both the OMP-AuNPs and PE-AuNPs exhibited a strong antioxidant potential in terms of 1,1-diphenyl-2-picrylhydraxyl radical scavenging, nitric oxide scavenging, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) radical scavenging, and a reducing power, along with a strong proteasome inhibitory potential that could be useful in cancer drug delivery and cancer treatments. The PE-AuNPs showed comparatively higher activity than OMP-AuNPs, which could be attributed to the presence of rich bioactive compounds in the PE that acted as reducing and capping agents in the synthesis of PE-AuNPs. Overall, the results of the current investigation highlighted a novel green technology for the synthesis of AuNPs using food waste materials and their potential applications in the biomedical, pharmaceutical, and cosmetic industries. PMID:27695326

Continuing global monitoring of anthropogenic and volcanic SO2 sources from Aura/OMI to SNPP/OMPS

NASA Astrophysics Data System (ADS)

Li, C.; Krotkov, N. A.; Carn, S. A.; Joiner, J.; McLinden, C. A.; Fioletov, V.

2017-12-01

Sulfur dioxide (SO2) is an important trace gas that has significant impacts on air quality, the climate, and stratospheric ozone. It is predominantly emitted from anthropogenic sources such as coal-fired power plants and smelters, but also has sizable sources from volcanic activity. The Ozone Monitoring Instrument (OMI) aboard NASA's Earth Observing System (EOS) Aura spacecraft has been providing global observations of both anthropogenic and volcanic SO2 since its launch in 2004. The Ozone Mapping and Profiler Suite (OMPS) nadir mapper, launched aboard the NASA/NOAA Suomi National Polar-orbiting Partnership (SNPP) satellite in 2011, will continue the 13+ year OMI SO2 time series. However, it is a significant challenge to build a coherent data record between OMI and OMPS, as the SO2 signal is relatively weak and its retrieval is subject to a number of interferences such as ozone absorption. Additionally, the two instruments also have different spectral and spatial resolutions that result in different sensitivities to SO2. Even a relatively small error in retrievals, due to either algorithmic or instrumental factors, may lead to a large inter-instrument bias in SO2. In this presentation, we report on our latest effort and progress in developing EOS continuity SNPP/OMPS SO2 products. We have applied a consistent retrieval technique based on principal component analysis (PCA) of measured radiance data to both OMI and OMPS. We show that the PCA retrieval technique, which extracts principal components (PCs) from measured radiances and applies these PCs in spectral fitting to minimize the effects of various interfering processes, is capable of producing highly consistent OMI and OMPS SO2 data for both anthropogenic sources and large volcanic eruptions. To gain a quantitative understanding of the two instruments' capabilities of monitoring SO2 sources, we have also applied a new emission estimation technique that combines satellite SO2 and wind information to both of them. This method allows SO2 emissions from 500 sources worldwide to be detected and quantified from OMI data. The OMPS-based emission estimates using the same method agree well with OMI-based ones, although OMPS detects fewer of the smaller sources seen by OMI, owing to its coarser spatial resolution.

Novel molecular markers of Chlamydia pecorum genetic diversity in the koala (Phascolarctos cinereus).

PubMed

Marsh, James; Kollipara, Avinash; Timms, Peter; Polkinghorne, Adam

2011-04-18

Chlamydia pecorum is an obligate intracellular bacterium and the causative agent of reproductive and ocular disease in several animal hosts including koalas, sheep, cattle and goats. C. pecorum strains detected in koalas are genetically diverse, raising interesting questions about the origin and transmission of this species within koala hosts. While the ompA gene remains the most widely-used target in C. pecorum typing studies, it is generally recognised that surface protein encoding genes are not suited for phylogenetic analysis and it is becoming increasingly apparent that the ompA gene locus is not congruent with the phylogeny of the C. pecorum genome. Using the recently sequenced C. pecorum genome sequence (E58), we analysed 10 genes, including ompA, to evaluate the use of ompA as a molecular marker in the study of koala C. pecorum genetic diversity. Three genes (incA, ORF663, tarP) were found to contain sufficient nucleotide diversity and discriminatory power for detailed analysis and were used, with ompA, to genotype 24 C. pecorum PCR-positive koala samples from four populations. The most robust representation of the phylogeny of these samples was achieved through concatenation of all four gene sequences, enabling the recreation of a "true" phylogenetic signal. OmpA and incA were of limited value as fine-detailed genetic markers as they were unable to confer accurate phylogenetic distinctions between samples. On the other hand, the tarP and ORF663 genes were identified as useful "neutral" and "contingency" markers respectively, to represent the broad evolutionary history and intra-species genetic diversity of koala C. pecorum. Furthermore, the concatenation of ompA, incA and ORF663 sequences highlighted the monophyletic nature of koala C. pecorum infections by demonstrating a single evolutionary trajectory for koala hosts that is distinct from that seen in non-koala hosts. While the continued use of ompA as a fine-detailed molecular marker for epidemiological analysis appears justified, the tarP and ORF663 genes also appear to be valuable markers of phylogenetic or biogeographic divisions at the C. pecorum intra-species level. This research has significant implications for future typing studies to understand the phylogeny, genetic diversity, and epidemiology of C. pecorum infections in the koala and other animal species.

Orbital-optimized MP2.5 and its analytic gradients: Approaching CCSD(T) quality for noncovalent interactions

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bozkaya, Uğur, E-mail: ugur.bozkaya@atauni.edu.tr; Center for Computational Molecular Science and Technology, School of Chemistry and Biochemistry, and School of Computational Science and Engineering, Georgia Institute of Technology, Atlanta, Georgia 30332; Sherrill, C. David

2014-11-28

Orbital-optimized MP2.5 [or simply “optimized MP2.5,” OMP2.5, for short] and its analytic energy gradients are presented. The cost of the presented method is as much as that of coupled-cluster singles and doubles (CCSD) [O(N{sup 6}) scaling] for energy computations. However, for analytic gradient computations the OMP2.5 method is only half as expensive as CCSD because there is no need to solve λ{sub 2}-amplitude equations for OMP2.5. The performance of the OMP2.5 method is compared with that of the standard second-order Møller–Plesset perturbation theory (MP2), MP2.5, CCSD, and coupled-cluster singles and doubles with perturbative triples (CCSD(T)) methods for equilibrium geometries, hydrogenmore » transfer reactions between radicals, and noncovalent interactions. For bond lengths of both closed and open-shell molecules, the OMP2.5 method improves upon MP2.5 and CCSD by 38%–43% and 31%–28%, respectively, with Dunning's cc-pCVQZ basis set. For complete basis set (CBS) predictions of hydrogen transfer reaction energies, the OMP2.5 method exhibits a substantially better performance than MP2.5, providing a mean absolute error of 1.1 kcal mol{sup −1}, which is more than 10 times lower than that of MP2.5 (11.8 kcal mol{sup −1}), and comparing to MP2 (14.6 kcal mol{sup −1}) there is a more than 12-fold reduction in errors. For noncovalent interaction energies (at CBS limits), the OMP2.5 method maintains the very good performance of MP2.5 for closed-shell systems, and for open-shell systems it significantly outperforms MP2.5 and CCSD, and approaches CCSD(T) quality. The MP2.5 errors decrease by a factor of 5 when the optimized orbitals are used for open-shell noncovalent interactions, and comparing to CCSD there is a more than 3-fold reduction in errors. Overall, the present application results indicate that the OMP2.5 method is very promising for open-shell noncovalent interactions and other chemical systems with difficult electronic structures.« less

Assembly of the β-Barrel Outer Membrane Proteins in Gram-Negative Bacteria, Mitochondria, and Chloroplasts

PubMed Central

Misra, Rajeev

2012-01-01

In the last decade, there has been an explosion of publications on the assembly of β-barrel outer membrane proteins (OMPs), which carry out diverse cellular functions, including solute transport, protein secretion, and assembly of protein and lipid components of the outer membrane. Of the three outer membrane model systems—Gram-negative bacteria, mitochondria and chloroplasts—research on bacterial and mitochondrial systems has so far led the way in dissecting the β-barrel OMP assembly pathways. Many exciting discoveries have been made, including the identification of β-barrel OMP assembly machineries in bacteria and mitochondria, and potentially the core assembly component in chloroplasts. The atomic structures of all five components of the bacterial β-barrel assembly machinery (BAM) complex, except the β-barrel domain of the core BamA protein, have been solved. Structures reveal that these proteins contain domains/motifs known to facilitate protein-protein interactions, which are at the heart of the assembly pathways. While structural information has been valuable, most of our current understanding of the β-barrel OMP assembly pathways has come from genetic, molecular biology, and biochemical analyses. This paper provides a comparative account of the β-barrel OMP assembly pathways in Gram-negative bacteria, mitochondria, and chloroplasts. PMID:27335668

Mapping the Laminin Receptor Binding Domains of Neisseria meningitidis PorA and Haemophilus influenzae OmpP2

PubMed Central

Mahdavi, Jafar; Oldfield, Neil J.; Wheldon, Lee M.; Wooldridge, Karl G.; Ala'Aldeen, Dlawer A. A.

2012-01-01

Neisseria meningitidis, Haemophilus influenzae and Streptococcus pneumoniae are major bacterial agents of meningitis. They each bind the 37/67-kDa laminin receptor (LamR) via the surface protein adhesins: meningococcal PilQ and PorA, H. influenzae OmpP2 and pneumococcal CbpA. We have previously reported that a surface-exposed loop of the R2 domain of CbpA mediates LamR-binding. Here we have identified the LamR-binding regions of PorA and OmpP2. Using truncated recombinant proteins we show that binding is dependent on amino acids 171–240 and 91–99 of PorA and OmpP2, respectively, which are predicted to localize to the fourth and second surface-exposed loops, respectively, of these proteins. Synthetic peptides corresponding to the loops bound LamR and could block LamR-binding to bacterial ligands in a dose dependant manner. Meningococci expressing PorA lacking the apex of loop 4 and H. influenzae expressing OmpP2 lacking the apex of loop 2 showed significantly reduced LamR binding. Since both loops are hyper-variable, our data may suggest a molecular basis for the range of LamR-binding capabilities previously reported among different meningococcal and H. influenzae strains. PMID:23049988

Yersinia pestis requires the 2-component regulatory system OmpR-EnvZ to resist innate immunity during the early and late stages of plague.

PubMed

Reboul, Angéline; Lemaître, Nadine; Titecat, Marie; Merchez, Maud; Deloison, Gaspard; Ricard, Isabelle; Pradel, Elizabeth; Marceau, Michaël; Sebbane, Florent

2014-11-01

Plague is transmitted by fleas or contaminated aerosols. To successfully produce disease, the causal agent (Yersinia pestis) must rapidly sense and respond to rapid variations in its environment. Here, we investigated the role of 2-component regulatory systems (2CSs) in plague because the latter are known to be key players in bacterial adaptation to environmental change. Along with the previously studied PhoP-PhoQ system, OmpR-EnvZ was the only one of Y. pestis' 23 other 2CSs required for production of bubonic, septicemic, and pneumonic plague. In vitro, OmpR-EnvZ was needed to counter serum complement and leukocytes but was not required for the secretion of antiphagocyte exotoxins. In vivo, Y. pestis lacking OmpR-EnvZ did not induce an early immune response in the skin and was fully virulent in neutropenic mice. We conclude that, throughout the course of Y. pestis infection, OmpR-EnvZ is required to counter toxic effectors secreted by polymorphonuclear leukocytes in the tissues. © The Author 2014. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

Mapping the laminin receptor binding domains of Neisseria meningitidis PorA and Haemophilus influenzae OmpP2.

PubMed

Abouseada, Noha M; Assafi, Mahde Saleh A; Mahdavi, Jafar; Oldfield, Neil J; Wheldon, Lee M; Wooldridge, Karl G; Ala'Aldeen, Dlawer A A

2012-01-01

Neisseria meningitidis, Haemophilus influenzae and Streptococcus pneumoniae are major bacterial agents of meningitis. They each bind the 37/67-kDa laminin receptor (LamR) via the surface protein adhesins: meningococcal PilQ and PorA, H. influenzae OmpP2 and pneumococcal CbpA. We have previously reported that a surface-exposed loop of the R2 domain of CbpA mediates LamR-binding. Here we have identified the LamR-binding regions of PorA and OmpP2. Using truncated recombinant proteins we show that binding is dependent on amino acids 171-240 and 91-99 of PorA and OmpP2, respectively, which are predicted to localize to the fourth and second surface-exposed loops, respectively, of these proteins. Synthetic peptides corresponding to the loops bound LamR and could block LamR-binding to bacterial ligands in a dose dependant manner. Meningococci expressing PorA lacking the apex of loop 4 and H. influenzae expressing OmpP2 lacking the apex of loop 2 showed significantly reduced LamR binding. Since both loops are hyper-variable, our data may suggest a molecular basis for the range of LamR-binding capabilities previously reported among different meningococcal and H. influenzae strains.

Pilot-scale study of powdered activated carbon recirculation for micropollutant removal.

PubMed

Meinel, F; Sperlich, A; Jekel, M

Adsorption onto powdered activated carbon (PAC) is a promising technique for the removal of organic micropollutants (OMPs) from treated wastewater. To enhance the adsorption efficiency, PAC is recycled back into the adsorption stage. This technique was examined in pilot scale in comparison to a reference without recirculation. Coagulation with Fe(3+) was carried out simultaneously to adsorption. Extensive OMP measurements showed that recirculation significantly increased OMP eliminations. Thus, significant PAC savings were feasible. The PAC concentration in the contact reactor proved to be an important operating parameter that can be surrogated by the easily measurable total suspended solids (TSS) concentration. OMP eliminations increased with increasing TSS concentrations. At 20 mg PAC L(-1) and 2.8 g TSS L(-1) in the contact reactor, well-adsorbable carbamazepine was eliminated by 97%, moderately adsorbable diclofenac was eliminated by 92% and poorly-adsorbable acesulfame was eliminated by 54% in comparison to 49%, 35% and 18%, respectively, without recirculation. The recirculation system represents an efficient technique, as the PAC's adsorption capacity is practically completely used. Small PAC dosages yield high OMP eliminations. Poorly-adsorbable gabapentin was eliminated to an unexpectedly high degree. A laboratory-scale biomass inhibition study showed that aerobic biodegradation removed gabapentin in addition to adsorption.

Evolutionary changes of the importance of olfaction in cetaceans based on the olfactory marker protein gene.

PubMed

Kishida, Takushi; Thewissen, J G M

2012-01-25

Odontocetes and mysticetes are two extant suborders of cetaceans. It is reported that the former have no sense of olfaction, while the latter can smell in air. To explain the ecological reason why mysticetes still retain their sense of smell, two hypotheses have been proposed - the echolocation-priority hypothesis, which assumes that the acquisition of echolocation causes the reduction of the importance of olfaction, and the filter-feeder hypothesis, which assumes that olfactory ability is important for filter-feeders to locate their prey because clouds of plankton give off a peculiar odor. The olfactory marker protein (OMP) is almost exclusively expressed in vertebrate olfactory receptor neurons, and is considered to play important roles in olfactory systems. In this study, full-length open reading frames of OMP genes were identified in 6 cetacean species and we analyzed the nonsynonymous to synonymous substitution rate ratio based on the maximum likelihood method. The evolutionary changes of the selective pressures on OMP genes did fit better to the filter-feeder hypothesis than to the echolocation-priority hypothesis. In addition, no pseudogenization mutations are found in all five odontocetes OMP genes investigated in this study. It may suggest that OMP retains some function even in 'anosmic' odontocetes. Copyright © 2011 Elsevier B.V. All rights reserved.

Th2-biased immune response and agglutinating antibodies generation by a chimeric protein comprising OmpC epitope (323-336) of Aeromonas hydrophila and LTB.

PubMed

Sharma, Mahima; Dash, Pujarini; Sahoo, Pramod K; Dixit, Aparna

2018-02-01

Aeromonas hydrophila is responsible for causing fatal infections in freshwater fishes. Besides chemical/antibiotic treatment and whole-cell vaccine, no subunit vaccine is currently available for A. hydrophila. Outer membrane proteins of gram-negative bacteria have been reported as effective vaccine candidates. Peptide antigens elicit focused immune responses against immunodominant stretches of the antigen. We have attempted to characterize the immunogenicity of linear B-cell epitopes of outer membrane protein (OmpC) of A. hydrophila identified using in silico tools, in conjugation with heat-labile enterotoxin B (LTB) subunit of Escherichia coli as a carrier protein. Antisera against the fusion protein harboring 323-336 residues of the AhOmpC (raised in mice) showed maximum cross-reactivity with the parent protein OmpC and LTB. The fusion protein displayed efficient GM 1 ganglioside receptor binding, retaining the adjuvanicity of LTB. Antibody isotype profile and in vitro T-cell response analysis, cytokine ELISA, and array analysis collectively revealed a Th2-biased mixed T-helper cell response. Agglutination assay and flow cytometry analysis validated the ability of anti-fusion protein antisera to recognize the surface exposed epitopes on Aeromonas cells, demonstrating its neutralization potential. Oral immunization studies in Labeo rohita resulted in the generation of long-lasting humoral immune response, and the antisera could cross-react with the fusion protein as well as both the fusion partners. Considering significant similarity among OmpC of different enteric bacteria, the use of A. hydrophila OmpC epitope 323-336 in fusion with LTB could have a broader scope in vaccine design.

The differential importance of mutations within AmpD in cephalosporin resistance of Enterobacter aerogenes and Enterobacter cloacae.

PubMed

Babouee Flury, Baharak; Ellington, Matthew J; Hopkins, Katie L; Turton, Jane F; Doumith, Michel; Woodford, Neil

2016-11-01

Mechanisms leading to carbapenem and cephalosporin resistance were sought in Enterobacter aerogenes isolates that were highly resistant to carbapenems but had no known carbapenemase. Results were compared with recent work examining carbapenem-resistant Enterobacter cloacae. Eighteen carbapenem-resistant E. aerogenes were screened for known β-lactamase and carbapenemase genes, and novel carbapenemases were sought in whole-genome sequencing (WGS) data of the three most resistant isolates. For all isolates, ampC, ampR, ampD and the porin genes omp35 and omp36 were investigated by Sanger sequencing or from available WGS data. Expression of ampC and porin genes was measured in comparison with cephalosporin- and carbapenem-susceptible control strains by reverse transcriptase PCR, with porin translation also detected by SDS-PAGE. Loss of Omp35, primarily due to decreased transcription (up to 250×), was observed in ertapenem-resistant isolates (MICs ≥ 2 mg/L), whereas meropenem resistance (MICs ≥ 4 mg/L) was observed in those isolates also showing decreased or no production of Omp36. Loss of Omp36 was due to combinations of premature translation termination or reduced transcription. In contrast to E. cloacae, cephalosporin resistance in E. aerogenes was not associated with lesions in AmpD. High-level cefepime resistance (MIC = 32 mg/L) was caused by a novel modification in the H-10 helix of AmpC in one isolate. The differential importance of AmpD lesions in cephalosporin resistance in E. cloacae and E. aerogenes underlines the differences between these contrasting members of the Enterobacter genus. Porin loss resulted in high-level carbapenem resistance with gradual loss of Omp36, which led to high-level meropenem resistance. Crown Copyright © 2016. Published by Elsevier B.V. All rights reserved.

VDAC electronics: 4. Novel electrical mechanism and thermodynamic estimations of glucose repression of yeast respiration.

PubMed

Lemeshko, Victor V

2017-11-01

Inhibition of cell respiration by high concentrations of glucose (glucose repression), known as "Crabtree effect", has been demonstrated for various cancerous strains, highly proliferating cells and yeast lines. Although significant progress in understanding metabolic events associated with the glucose repression of cell respiration has been achieved, it is not yet clear whether the Crabtree effect is the result of a limited activity of the respiratory chain, or of some glucose-mediated regulation of mitochondrial metabolic state. In this work we propose an electrical mechanism of glucose repression of the yeast S. cerevisiae, resulting from generation of the mitochondrial outer membrane potential (OMP) coupled to the direct oxidation of cytosolic NADH in mitochondria. This yeast-type mechanism of OMP generation is different from the earlier proposed VDAC-hexokinase-mediated voltage generation of cancer-type, associated with the mitochondrial outer membrane. The model was developed assuming that VDAC is more permeable to NADH than to NAD + . Thermodynamic estimations of OMP, generated as a result of NADH(2-)/NAD + (1-) turnover through the outer membrane, demonstrated that the values of calculated negative OMP match the known range of VDAC voltage sensitivity, thus suggesting a possibility of OMP-dependent VDAC-mediated regulation of cell energy metabolism. According to the proposed mechanism, we suggest that the yeast-type Crabtree effect is the result of a fast VDAC-mediated electrical repression of mitochondria due to a decrease in the outer membrane permeability to charged metabolites and owing their redistribution between the mitochondrial intermembrane space and the cytosol, both controlled by metabolically-derived OMP. Copyright © 2017 Elsevier B.V. All rights reserved.

Population-based genetic epidemiologic analysis of Chlamydia trachomatis serotypes and lack of association between ompA polymorphisms and clinical phenotypes.

PubMed

Millman, Kim; Black, Carolyn M; Stamm, Walter E; Jones, Robert B; Hook, Edward W; Martin, David H; Bolan, Gail; Tavaré, Simon; Dean, Deborah

2006-03-01

Chlamydia trachomatis is the leading cause of bacterial sexually transmitted diseases worldwide. Urogenital strains are classified into serotypes and genotypes based on the major outer membrane protein and its gene, ompA, respectively. Studies of the association of serotypes with clinical signs and symptoms have produced conflicting results while no studies have evaluated associations with ompA polymorphisms. We designed a population-based cross-sectional study of 344 men and women with urogenital chlamydial infections (excluding co-pathogen infections) presenting to clinics serving five U.S. cities from 1995 to 1997. Signs, symptoms and sequelae of chlamydial infection (mucopurulent cervicitis, vaginal or urethral discharge; dysuria; lower abdominal pain; abnormal vaginal bleeding; and pelvic inflammatory disease) were analyzed for associations with serotype and ompA polymorphisms. One hundred and fifty-three (44.5%) of 344 patients had symptoms consistent with urogenital chlamydial infection. Gender, reason for visit and city were significant independent predictors of symptom status. Men were 2.2 times more likely than women to report any symptoms (P=0.03) and 2.8 times more likely to report a urethral discharge than women were to report a vaginal discharge in adjusted analyses (P=0.007). Differences in serotype or ompA were not predictive except for an association between serotype F and pelvic inflammatory disease (P=0.046); however, the number of these cases was small. While there was no clinically prognostic value associated with serotype or ompA polymorphism for urogenital chlamydial infections except for serotype F, future studies might utilize multilocus genomic typing to identify chlamydial strains associated with clinical phenotypes.

VDAC electronics: 3. VDAC-Creatine kinase-dependent generation of the outer membrane potential in respiring mitochondria.

PubMed

Lemeshko, Victor V

2016-07-01

Mitochondrial energy in cardiac cells has been reported to be channeled into the cytosol through the intermembrane contact sites formed by the adenine nucleotide translocator, creatine kinase and VDAC. Computational analysis performed in this study showed a high probability of the outer membrane potential (OMP) generation coupled to such a mechanism of energy channeling in respiring mitochondria. OMPs, positive inside, calculated at elevated concentrations of creatine are high enough to restrict ATP release from mitochondria, to significantly decrease the apparent K(m,ADP) for state 3 respiration and to maintain low concentrations of Ca(2+) in the mitochondrial intermembrane space. An inhibition by creatine of Ca(2+)-induced swelling of isolated mitochondria and other protective effects of creatine reported in the literature might be explained by generated positive OMP. We suggest that VDAC-creatine kinase-dependent generation of OMP represents a novel physiological factor controlling metabolic state of mitochondria, cell energy channeling and resistance to death. Copyright © 2016 Elsevier B.V. All rights reserved.

BIODEGRADATION OF ORGANOPHOSPHORUS PESTICIDES BY SURFACE-EXPRESSED ORGANOPHOSPHORUS HYDROLASE. (R823663)

EPA Science Inventory

Organophosphorus hydrolase (OPH) was displayed and anchored onto the surface of
Escherichia coli using an Lpp-OmpA fusion system. Production of the fusion proteins in membrane
fractions was verified by immunoblotting with OmpA antisera. inclusion of the organophosphorus
...

User interface issues in supporting human-computer integrated scheduling

NASA Technical Reports Server (NTRS)

Cooper, Lynne P.; Biefeld, Eric W.

1991-01-01

The topics are presented in view graph form and include the following: characteristics of Operations Mission Planner (OMP) schedule domain; OMP architecture; definition of a schedule; user interface dimensions; functional distribution; types of users; interpreting user interaction; dynamic overlays; reactive scheduling; and transitioning the interface.

Survival of mature mouse olfactory sensory neurons labeled genetically perinatally.

PubMed

Holl, Anna-Maria

2018-04-01

The main olfactory epithelium (MOE) of an adult mouse harbors a few million mature olfactory sensory neurons (OSNs), which are traditionally defined as mature by their expression of the olfactory marker protein (OMP). Mature OSNs differentiate in situ from stem cells at the base of the MOE. The consensus view is that mature OSNs have a defined lifespan and then undergo programmed cell death, and that the adult MOE maintains homeostasis by generating new mature OSNs from stem cells. But there is also evidence for mature OSNs that are long-lived. Thus far modern genetic tools have not been applied to quantify survival of a population of OSNs that are mature at a given point in time. Here, a genetic strategy was developed to label irreversibly OMP-expressing OSNs in mice. A gene-targeted OMP-CreERT2 strain was generated in which mature OSNs express an enzymatically inactive version of the Cre recombinase. The fusion protein CreERT2 becomes transiently active when exposed to tamoxifen, and in the presence of a Cre reporter in the genome such as tdRFP, CreERT2-expressing cells become irreversibly labeled. A cohort of mice was generated with the same day of birth by in vitro fertilization and embryo transfer, and injected tamoxifen in their mothers at E18.5 of gestation. I counted RFP immunoreactive cells in the MOE and vomeronasal organ of 36 tamoxifen-exposed OMP-CreERT2 × tdRFP mice from 7 age groups: postnatal day (PD)1.5, PD3.5, PD6.5, 3 weeks, 9 weeks, 6 months, and 12 months. Approximately 7.8% of perinatally labeled cells remain at 12 months, confirming that some mature OSNs are indeed long-lived. The survival curve of the population of perinatally labeled MOE cells can be modeled with a mean half-life of 26 days for the population as a whole, excluding the long-lived cells. Copyright © 2018 The Author. Published by Elsevier Inc. All rights reserved.

Assessment of OmpATb as a Novel Antigen for the Diagnosis of Bovine Tuberculosis

USDA-ARS?s Scientific Manuscript database

In search for better tools to control bovine tuberculosis, the development of diagnostic tests with improved specificity and sensitivity has a high priority. We chose to search for novel immunodiagnostic reagents. In this study, Rv0899 (Outer membrane protein A of Mycobacterium tuberculosis, OmpATb)...

Determinants for successful marketing authorisation of orphan medicinal products in the EU.

PubMed

Putzeist, Michelle; Heemstra, Harald E; Garcia, Jordi Llinares; Mantel-Teeuwisse, Aukje K; Gispen-De Wied, Christine C; Hoes, Arno W; Leufkens, Hubert G M

2012-04-01

In 2010, the European Regulation for Orphan Medicinal Products (OMPs) was in force for ten years. In this study we assessed possible determinants of applications for OMPs in the EU since 2000 that are associated with a successful marketing authorisation. Our analysis shows that clinical trial characteristics such as demonstrating convincing evidence of a beneficial effect on the primary endpoint, the selection of a clinically relevant endpoint, providing RCT data as pivotal study evidence and the submission of sound dose finding data are critical success factors. In addition, high medical need seems to counterweigh uncertainties about the scientific evidence in the benefit-risk assessment of OMPs. Copyright © 2011 Elsevier Ltd. All rights reserved.

Development of the Multi-Angle Stratospheric Aerosol Radiometer (MASTAR) Instrument

NASA Astrophysics Data System (ADS)

DeLand, M. T.; Colarco, P. R.; Kowalewski, M. G.; Gorkavyi, N.; Ramos-Izquierdo, L.

2017-12-01

Aerosol particles in the stratosphere ( 15-25 km altitude), both produced naturally and perturbed by volcanic eruptions and anthropogenic emissions, continue to be a source of significant uncertainty in the Earth's energy budget. Stratospheric aerosols can offset some of the warming effects caused by greenhouse gases. These aerosols are currently monitored using measurements from the Ozone Mapping and Profiling Suite (OMPS) Limb Profiler (LP) instrument on the Suomi NPP satellite. In order to improve the sensitivity and spatial coverage of these aerosol data, we are developing an aerosol-focused compact version of the OMPS LP sensor called Multi-Angle Stratospheric Aerosol Radiometer (MASTAR) to fly on a 3U Cubesat satellite, using a NASA Instrument Incubator Program (IIP) grant. This instrument will make limb viewing measurements of the atmosphere in multiple directions simultaneously, and uses only a few selected wavelengths to reduce size and cost. An initial prototype version has been constructed using NASA GSFC internal funding and tested in the laboratory. Current design work is targeted towards a preliminary field test in Spring 2018. We will discuss the scientific benefits of MASTAR and the status of the project.

Backbone ¹H, ¹³C, ¹⁵N NMR assignments of yeast OMP synthase in unliganded form and in complex with orotidine 5'-monophosphate.

PubMed

Hansen, Michael Riis; Harris, Richard; Barr, Eric W; Cheng, Hong; Girvin, Mark E; Grubmeyer, Charles

2014-04-01

The type I phosphoribosyltransferase OMP synthase (EC 2.4.2.10) is involved in de novo synthesis of pyrimidine nucleotides forming the UMP precursor orotidine 5'-monophosphate (OMP). The homodimeric enzyme has a Rossman α/β core topped by a base-enclosing "hood" domain and a flexible domain-swapped catalytic loop. High-resolution X-ray structures of the homologous Salmonella typhimurium and yeast enzymes show that a general compacting of the core as well as movement of the hood and a major disorder-to-order transition of the loop occur upon binding of ligands MgPRPP and orotate. Here we present backbone NMR assignments for the unliganded yeast enzyme (49 kDa) and its complex with product OMP. We were able to assign 212-213 of the 225 non-proline backbone (15)N and amide proton resonances. Significant difference in chemical shifts of the amide cross peaks occur in regions of the structure that undergo movement upon ligand occupancy in the S. typhimurium enzyme.

Merged SAGE II / MIPAS / OMPS Ozone Record : Impact of Transfer Standard on Ozone Trends.

NASA Astrophysics Data System (ADS)

Kramarova, N. A.; Laeng, A.; von Clarmann, T.; Stiller, G. P.; Walker, K. A.; Zawodny, J. M.; Plieninger, J.

2017-12-01

The deseasonalized ozone anomalies from SAGE II, MIPAS and OMPS-LP datasets are merged into one long record. Two versions of the dataset will be presented : ACE-FTS instrument or MLS instrument are used as a transfer standard. The data are provided in 10 degrees latitude bins, going from 60N to 60S for the period from October 1984 to March 2017. The main differences between presented in this study merged ozone record and the merged SAGE II / Ozone_CCI / OMPS-Saskatoon dataset by V. Sofieva are: - the OMPS-LP data are from the NASA GSFC version 2 processor - the MIPAS 2002-2004 date are taken into the record - Data are merged using a transfer standard. In overlapping periods data are merged as weighted means where the weights are inversely proportional to the standard errors of the means (SEM) of the corresponding individual monthly means. The merged dataset comes with the uncertainty estimates. Ozone trends are calculated out of both versions of the dataset. The impact of transfer standard on obtained trends is discussed.

Rapid small-scale column testing of granular activated carbon for organic micro-pollutant removal in treated domestic wastewater.

PubMed

Zietzschmann, F; Müller, J; Sperlich, A; Ruhl, A S; Meinel, F; Altmann, J; Jekel, M

2014-01-01

This study investigates the applicability of the rapid small-scale column test (RSSCT) concept for testing of granular activated carbon (GAC) for organic micro-pollutants (OMPs) removal from wastewater treatment plant (WWTP) effluent. The chosen experimental setup was checked using pure water, WWTP effluent, different GAC products, and variable hydrodynamic conditions with different flow velocities and differently sized GAC, as well as different empty bed contact times (EBCTs). The setup results in satisfying reproducibility and robustness. RSSCTs in combination with WWTP effluent are effective when comparing the OMP removal potentials of different GAC products and are a useful tool for the estimation of larger filters. Due to the potentially high competition between OMPs and bulk organics, breakthrough curves are likely to have unfavorable shapes when treating WWTP effluent. This effect can be counteracted by extending the EBCT. With respect to the strong competition observed in GAC treatment of WWTP effluent, the small organic acid and neutral substances are retained longer in the RSSCT filters and are likely to cause the majority of the observed adsorption competition with OMPs.

Omp85 genosensor for detection of human brain bacterial meningitis.

PubMed

Dash, Sandip Kumar; Sharma, Minakshi; Khare, Shashi; Kumar, Ashok

2013-06-01

The 5'-thiolated DNA probe based on specific virulence gene, Omp85, was immobilized onto a screen-printed gold electrode followed by hybridization with 6-100 ng/6 μl (5.9 × 10(5)-9.3 × 10(6 )c.f.u.) of Neisseria meningitidis single stranded genomic DNA (ssG-DNA) for 10 min at 25 °C from the cerebrospinal fluid (CSF) of a meningitis patient. The Omp85 genosensor can detect as little as 6 ng ssG-DNA in 6 μl CSF of a human brain meningitis patient in 30 min including a response time of 1 min by cyclic voltammetry, differential pulse voltammetry (DPV) and electrochemical impedance. The sensitivity of the genosensor electrode was 2.6(μA/cm(2))/ng using DPV with regression coefficient (R(2)) 0.954. The genosensor was characterized using Fourier transform infrared spectroscopy and atomic force microscopy. Omp85 genosensor was stable for 12 months at 4 °C with 12 % loss in DPV current.

16S rRNA and Omp31 Gene Based Molecular Characterization of Field Strains of B. melitensis from Aborted Foetus of Goats in India

PubMed Central

Singh, Ajay; Gupta, Vivek Kumar; Kumar, Amit; Singh, Vikas Kumar; Nayakwadi, Shivasharanappa

2013-01-01

Brucellosis is a reemerging infectious zoonotic disease of worldwide importance. In human, it is mainly caused by Brucella melitensis, a natural pathogen for goats. In India, a large number of goats are reared in semi-intensive to intensive system within the close vicinity of human being. At present, there is no vaccination and control strategy for caprine brucellosis in the country. Thus, to formulate an effective control strategy, the status of etiological agent is essential. To cope up with these, the present study was conducted to isolate and identify the prevalent Brucella species in caprine brucellosis in India. The 30 samples (fetal membrane, fetal stomach content and vaginal swabs) collected throughout India from the aborted fetus of goats revealed the isolation of 05 isolates all belonging to Brucella melitensis biovars 3. All the isolates produced amplification products of 1412 and 720 bp in polymerase chain reaction with genus and species specific 16S rRNA and omp31 gene based primers, respectively. Moreover, the amplification of omp31 gene in all the isolates confirmed the presence of immuno dominant outer membrane protein (31 kDa omp) in all the field isolates of B. melitensis in aborted foetus of goats in India. These findings can support the development of omp31 based specific serodiagnostic test as well as vaccine for the control of caprine brucellosis in India. PMID:24453799

Identification of minimum carbohydrate moiety in N-glycosylation sites of brain endothelial cell glycoprotein 96 for interaction with Escherichia coli K1 outer membrane protein A

PubMed Central

Krishnan, Subramanian; Prasadarao, Nemani V.

2014-01-01

Bacterial meningitis is a serious central nervous system infection and Escherichia coli K1 (E. coli K1) is one of the leading etiological agents that cause meningitis in neonates. Outer membrane protein A (OmpA) of E. coli K1 is a major virulence factor in the pathogenesis of meningitis, and interacts with human brain microvascular endothelial cells (HBMEC) to cross the blood-brain barrier. Using site-directed mutagenesis, we demonstrate that two N-glycosylation sites (NG1 and NG2) in the extracellular domain of OmpA receptor, Ecgp96 are critical for bacterial binding to HBMEC. E. coli invasion assays using CHO-Lec1 cells that express truncated N-glycans, and sequential digestion of HBMEC surface N-glycans using specific glycosidases showed that GlcNAc1-4GlcNAc epitopes are sufficient for OmpA interaction with HBMEC. Lack of NG1 and NG2 sites in Ecgp96 inhibits E. coli OmpA induced F-actin polymerization, phosphorylation of protein kinase C-α, and disruption of transendothelial electrical resistance required for efficient invasion of E. coli in HBMEC. Furthermore, the microvessels of cortex and hippocampus of the brain sections of E. coli K1 infected mice showed increased expression of glycosylated Ecgp96. Therefore, the interface of OmpA and GlcNAc1-4GlcNAc epitope interaction would be a target for preventative strategies against E. coli K1 meningitis. PMID:24932957

'Candidatus Rickettsia nicoyana': A novel Rickettsia species isolated from Ornithodoros knoxjonesi in Costa Rica.

PubMed

Moreira-Soto, Rolando D; Moreira-Soto, Andrés; Corrales-Aguilar, Eugenia; Calderón-Arguedas, Ólger; Troyo, Adriana

2017-06-01

Rickettsiae are intracellular bacteria commonly associated with hematophagous arthropods. Most of them have been described in hard ticks, but some have been found in soft ticks. Here we report the detection and isolation of a new Rickettsia from Ornithodoros knoxjonesi larvae collected from Balantiopteryx plicata (Emballonuridae) in Nicoya, Costa Rica. Two ticks were processed to detect Rickettsia spp. genes gltA, ompA, ompB, and htrA by PCR. Part of the macerate was also inoculated into Vero E6 and C6/36 cell lines, and cells were evaluated by Giménez stain, indirect immunofluorescence assay (IFA), and PCR. Both ticks were positive by PCR and rickettsial growth was successful in Vero E6 cells. Amplification and sequencing of near full length rrs, gltA, sca4 genes, and fragments of ompA and ompB showed that the Rickettsia sp. was different from described species. The highest homologies were with 'Candidatus Rickettsia wissemanii' and Rickettsia peacockii: 99.70% (1321/1325) with both sequences for rrs, 99.58% (1172/1177) and 99.76% (1246/1249) for gltA, 99.26% with both sequences (2948/2970 and 2957/2979) for sca4, 98.78% (485/491) and 98.39% (2069/2115) for ompA, and 98.58 (1453/1474) and 98.92% (1459/1475) for ompB; respectively. Bat blood, spleen, liver, and lung samples analyzed for Rickettsia detection were negative. Results demonstrate that the Rickettsia isolated from O. knoxjonesi is probably an undescribed species that belongs to the spotted fever group, for which 'Candidatus Rickettsia nicoyana' is proposed. Considering that B. plicata inhabits areas where contact with humans may occur and that human parasitism by Ornithodoros has been reported in the country, it will be important to continue with the characterization of this species and its pathogenic potential. Copyright © 2017 Elsevier GmbH. All rights reserved.

Concordance in Anti-OmpC and Anti-I2 Indicate the Influence of Genetic Predisposition: Results of a European Study of Twins with Crohn's Disease.

PubMed

Amcoff, Karin; Joossens, Marie; Pierik, Marie J; Jonkers, Daisy; Bohr, Johan; Joossens, Sofie; Romberg-Camps, Mariëlle; Nyhlin, Nils; Wickbom, Anna; Rutgeerts, Paul J; Tysk, Curt; Bodin, Lennart; Colombel, Jean-Frederic; Vermeire, Severine; Halfvarson, Jonas

2016-06-01

An adaptive immunological response to microbial antigens has been observed in Crohn's disease (CD). Intriguingly, this serological response precedes the diagnosis in some patients and has also been observed in healthy relatives. We aimed to determine whether genetic factors are implicated in this response in a CD twin cohort. In total, 82 twin pairs (Leuven n = 13, Maastricht n = 8, Örebro n = 61) took part: 81 pairs with CD (concordant monozygotic n = 16, discordant monozygotic n = 22, concordant dizygotic n = 3, discordant dizygotic n = 40) and 1 monozygotic pair with both CD and ulcerative colitis. Serology for Pseudomonas fluorescens-related protein (anti-I2), Escherichia coli outer membrane porin C (anti-OmpC), CBir1flagellin (anti-CBir1) and antibodies to oligomannan (anti-Saccharomyces cerevisiae antibody [ASCA]) was determined by standardized enzyme-linked immunoassay. All markers were more often present in CD twins than in their healthy twin siblings. Using the intraclass correlation coefficient (ICC), agreements in concentrations of anti-OmpC and anti-I2 were observed in discordant monozygotic but not in discordant dizygotic twin pairs with CD (anti-OmpC, ICC 0.80 and -0.02, respectively) and (anti-I2, ICC 0.56 and 0.05, respectively). In contrast, no agreements were found in anti-CBir, immunoglobulin (Ig) G ASCA and ASCA IgA. We show that anti-I2 and anti-CBir1 statuses have specificity for CD and confirm previous reported specificities for anti-OmpC and ASCA. Based on quantitative analyses and observed ICCs, genetics seems to predispose to the anti-OmpC and anti-I2 response but less to ASCA and anti-CBir1 responses. Copyright © 2016 European Crohn’s and Colitis Organisation (ECCO). Published by Oxford University Press. All rights reserved. For permissions, please email: journals.permissions@oup.com.

Validation of Suomi NPP OMPS Limb Profiler Ozone Measurements

NASA Astrophysics Data System (ADS)

Buckner, S. N.; Flynn, L. E.; McCormick, M. P.; Anderson, J.

2017-12-01

The Ozone Mapping and Profiler Suite (OMPS) Limb Profiler onboard the Suomi National Polar-Orbiting Partnership satellite (SNPP) makes measurements of limb-scattered solar radiances over Ultraviolet and Visible wavelengths. These measurements are used in retrieval algorithms to create high vertical resolution ozone profiles, helping monitor the evolution of the atmospheric ozone layer. NOAA is in the process of implementing these algorithms to make near-real-time versions of these products. The main objective of this project is to generate estimates of the accuracy and precision of the OMPS Limb products by analysis of matchup comparisons with similar products from the Earth Observing System Microwave Limb Sounder (EOS Aura MLS). The studies investigated the sources of errors, and classified them with respect to height, geographic location, and atmospheric and observation conditions. In addition, this project included working with the algorithm developers in an attempt to develop corrections and adjustments. Collocation and zonal mean comparisons were made and statistics were gathered on both a daily and monthly basis encompassing the entire OMPS data record. This validation effort of the OMPS-LP data will be used to help validate data from the Stratosphere Aerosol and Gas Experiment III on the International Space Station (SAGE III ISS) and will also be used in conjunction with the NOAA Total Ozone from Assimilation of Stratosphere and Troposphere (TOAST) product to develop a new a-priori for the NOAA Unique Combined Atmosphere Processing System (NUCAPS) ozone product. The current NUCAPS ozone product uses a combination of Cross-track Infrared Sounder (CrIS) data for the troposphere and a tropopause based climatology derived from ozonesonde data for the stratosphere a-priori. The latest version of TOAST uses a combination of both CrIS and OMPS-LP data. We will further develop the newest version of TOAST and incorporate it into the NUCAPS system as a new a-priori, in hopes of creating a better global ozone product.

Highly diverse MLVA-ompA genotypes of rectal Chlamydia trachomatis among men who have sex with men in Brighton, UK and evidence for an HIV-related sexual network.

PubMed

Labiran, Clare; Marsh, Peter; Zhou, Judith; Bannister, Alan; Clarke, Ian Nicholas; Goubet, Stephanie; Soni, Suneeta

2016-06-01

In this prospective study, we aimed to determine the distribution of genotypes by multilocus variable number tandem repeat (VNTR) analysis plus analysis of the ompA gene (MLVA-ompA) of rectal Chlamydia trachomatis among men who have sex with men (MSM) attending Brighton Genitourinary Medicine (GUM) Clinic and to examine any correlations with clinical variables, including HIV status, and to isolate rectal C. trachomatis cultures maximising the possibility of obtaining complete genotyping data. Samples were assigned genotypes by PCR and sequencing of the markers of the MLVA-ompA genotyping system. Rectal C. trachomatis was isolated in cell culture using McCoy cells. Data regarding demographics, HIV status, rectal symptoms and history of sexually transmitted infections, including C. trachomatis, were collected. 1809 MSM attending the clinic between October 2011 and January 2013 took part in the study, 112 (6.2%) of whom had rectal samples that tested positive for C. trachomatis. 85/112 (75.9%) C. trachomatis-positive rectal samples were assigned 66 different genotypes. Two distinct genotype subclusters were identified: subcluster 1 consisted of more HIV-negative men than subcluster 2 (p=0.025), and the MLVA-ompA genotypes in these subclusters reflected this. Isolates were successfully cultured from 37 of the 112 specimens, from which 27 otherwise unobtainable (from direct PCR) MLVA-ompA genotypes were gained. The most prevalent genotypes were G, E and D representing some overlap with the heterosexual distribution in UK. Subcluster 1 consisted of more 'heterosexual genotypes' and significantly more HIV-negative men than subcluster 2, associated with 'MSM genotypes'. There was a higher diversity of C. trachomatis strains among MSM in Brighton than observed in other cities. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/

Organic micro-pollutants in a karst system during low and high flow periods, results of a two-year study

NASA Astrophysics Data System (ADS)

Reh, Roland; Licha, Tobias; Geyer, Tobias; Nödler, Karsten; Sauter, Martin

2013-04-01

Fifty-four different organic micro-pollutants (OMPs) including pharmaceuticals, pesticides, corrosion inhibitors and other typical wastewater compounds such as caffeine are repeatedly analyzed in approximately fifty groundwater observation points in a complex faulted and fractured carbonate aquifer system consisting of three main spring catchment areas. The study area is characterized as well by the influence of an urban area with industrial sites and waste disposals as by surrounding agricultural uses. With the applied HPLC-MS/MS method (Nödler et al., 2010), achieving method quantification limits (MQL) of 1.8 - 28 ng L-1, forty-four of the OMPs are detected in groundwater. Indicating a subsurface water divide, OMPs distinguish between the North and the South main spring areas with respect to the number of detected OMPs, OMP combinations, occurrence of special compounds like tamoxifen and concentrations of triazoles and triazines. Regarding the vertical distribution in the aquifer system upper aquifers exhibit a higher number and higher seasonal variety of OMPs than the lower aquifer and the main spring areas. In addition to triazines and triazoles the anti-epileptic drug carbamazepine and the analgesic drug phenazone are the most frequently detected compounds. The detection of the estrogen antagonist tamoxifen in a groundwater flow system is reported for the first time. Defined combinations of OMPs occur repeatedly in the same observation wells and allow to distinguish different input functions. The comparison of detection frequency with the number of prescribed doses gives information about the specific persistance of pharmaceuticals. A disused waste disposal is identified as input area for triazines. Additional input of triazines in the agricultural areas may be indicated by different fractions of atrazine and its degradation products. Due to prohibition of atrazine as herbicide since 1992 the occurrence of OMPs in the lower aquifer, solely limited to atrazine and desethylatrazine, indicates limited aquifer connections and demonstrates a residence time of several years for vertical transport into the deep aquifer. Phenazone originates also from a disused waste disposal and is showing a persistency for more than three decades under the observed redox conditions. References Nödler K, Licha T, Bester K, Sauter M. Development of a multi-residue analytical method, based on liquid chromatography-tandem mass spectrometry, for the simultaneous determination of 46 micro-contaminants in aqueous samples. Journal of Chromatography A 2010;1217(42), 6511-6521. Reh R, Licha T, Geyer T, Nödler, K.,Sauter M. Occurrence and spatial distribution of organic micro-pollutants in a complex hydrogeological karst system during low flow and high flow periods, results of a two-year study. Science of The Total Environment 2013, 443,438-445, 10.1016/j.scitotenv.2012.11.005.

Rickettsia asembonensis Characterization by Multilocus Sequence Typing of Complete Genes, Peru.

PubMed

Loyola, Steev; Flores-Mendoza, Carmen; Torre, Armando; Kocher, Claudine; Melendrez, Melanie; Luce-Fedrow, Alison; Maina, Alice N; Richards, Allen L; Leguia, Mariana

2018-05-01

While studying rickettsial infections in Peru, we detected Rickettsia asembonensis in fleas from domestic animals. We characterized 5 complete genomic regions (17kDa, gltA, ompA, ompB, and sca4) and conducted multilocus sequence typing and phylogenetic analyses. The molecular isolate from Peru is distinct from the original R. asembonensis strain from Kenya.

Nucleotide and phylogenetic analyses of the Chlamydia trachomatis ompA gene indicates it is a hotspot for mutation

USDA-ARS?s Scientific Manuscript database

Background: Serovars of the human pathogen Chlamydia trachomatis occupy one of three specific tissue niches. Genomic analyses indicate that the serovars have a phylogeny congruent with their pathobiology and have an average substitution rate of less than one nucleotide per kilobase. The ompA gene, h...

Rickettsia parkeri in Dermacentor parumapertus Ticks, Mexico

PubMed Central

Sánchez-Montes, Sokani; Guzmán-Cornejo, Carmen; Colunga-Salas, Pablo; Becker, Ingeborg; Delgado-de la Mora, Jesús; Licona-Enríquez, Jesús D.; Delgado-de la Mora, David; Karpathy, Sandor E.; Paddock, Christopher D.; Suzán, Gerardo

2018-01-01

During a study to identify zoonotic pathogens in northwestern Mexico, we detected the presence of a rickettsial agent in Dermacentor parumapertus ticks from black-tailed jackrabbits (Lepus californicus). Comparison of 4 gene sequences (gltA, htrA, ompA, and ompB) of this agent showed 99%–100% identity with sequences of Rickettsia parkeri. PMID:29774838

Understanding the influence of surface chemical states on the dielectric tunability of sputtered Ba0.5Sr0.5TiO3 thin films

NASA Astrophysics Data System (ADS)

Venkata Saravanan, K.; Raju, K. C. James

2014-03-01

The surface chemical states of RF-magnetron sputtered Ba0.5Sr0.5TiO3 (BST5) thin films deposited at different oxygen mixing percentage (OMP) was examined by x-ray photoelectron spectroscopy. The O1s XPS spectra indicate the existence of three kinds of oxygen species (dissociated oxygen ion O2 -, adsorbed oxide ion O- and lattice oxide ion O2-) on the films’ surface, which strongly depends on OMP. The presence of oxygen species other than lattice oxygen ion makes the films’ surface highly reactivity to atmospheric gases, resulting in the formation of undesired surface layers. The XPS results confirm the formation of surface nitrates for the films deposited under oxygen deficient atmosphere (OMP ≦̸ 25%), whereas the films deposited in oxygen rich atmosphere (OMP ≧̸ 75%) show the presence of metal-hydroxide. The influence of a surface dead layer on the tunable dielectric properties of BST5 films have been studied in detail and are reported. Furthermore, our observations indicate that an optimum ratio of Ar:O2 is essential for achieving desired material and dielectric properties in BST5 thin films. The films deposited at 50% OMP have the highest dielectric tunability of ~65% (@280 kV cm-1), with good ɛ r-E curve symmetry of 98% and low tan δ of 0.018. The figure of merit for these films is about 35, which is promising for frequency agile device applications.

Host adaptation of Chlamydia pecorum towards low virulence evident in co-evolution of the ompA, incA, and ORF663 Loci.

PubMed

Mohamad, Khalil Yousef; Kaltenboeck, Bernhard; Rahman, Kh Shamsur; Magnino, Simone; Sachse, Konrad; Rodolakis, Annie

2014-01-01

Chlamydia (C.) pecorum, an obligate intracellular bacterium, may cause severe diseases in ruminants, swine and koalas, although asymptomatic infections are the norm. Recently, we identified genetic polymorphisms in the ompA, incA and ORF663 genes that potentially differentiate between high-virulence C. pecorum isolates from diseased animals and low-virulence isolates from asymptomatic animals. Here, we expand these findings by including additional ruminant, swine, and koala strains. Coding tandem repeats (CTRs) at the incA locus encoded a variable number of repeats of APA or AGA amino acid motifs. Addition of any non-APA/AGA repeat motif, such as APEVPA, APAVPA, APE, or APAPE, associated with low virulence (P<10-4), as did a high number of amino acids in all incA CTRs (P = 0.0028). In ORF663, high numbers of 15-mer CTRs correlated with low virulence (P = 0.0001). Correction for ompA phylogram position in ORF663 and incA abolished the correlation between genetic changes and virulence, demonstrating co-evolution of ompA, incA, and ORF663 towards low virulence. Pairwise divergence of ompA, incA, and ORF663 among isolates from healthy animals was significantly higher than among strains isolated from diseased animals (P≤10-5), confirming the longer evolutionary path traversed by low-virulence strains. All three markers combined identified 43 unique strains and 4 pairs of identical strains among all 57 isolates tested, demonstrating the suitability of these markers for epidemiological investigations.

Structure of thrombospondin type 3 repeats in bacterial outer membrane protein A reveals its intra-repeat disulfide bond-dependent calcium-binding capability

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dai, Shuyan; Sun, Cancan; Tan, Kemin

Eukaryotic thrombospondin type 3 repeat (TT3R) is an efficient calcium ion (Ca2+) binding motif only found in mammalian thrombospondin family. TT3R has also been found in prokaryotic cellulase Cel5G, which was thought to forfeit the Ca2+-binding capability due to the formation of intra-repeat disulfide bonds, instead of the inter-repeat ones possessed by eukaryotic TT3Rs. In this study, we have identified an enormous number of prokaryotic TT3R-containing proteins belonging to several different protein families, including outer membrane protein A (OmpA), an important structural protein connecting the outer membrane and the periplasmic peptidoglycan layer in gram-negative bacteria. Here, we report the crystalmore » structure of the periplasmic region of OmpA from Capnocytophaga gingivalis, which contains a linker region comprising five consecutive TT3Rs. The structure of OmpA-TT3R exhibits a well-ordered architecture organized around two tightly-coordinated Ca2+ and confirms the presence of abnormal intra-repeat disulfide bonds. Further mutagenesis studies showed that the Ca2+-binding capability of OmpA-TT3R is indeed dependent on the proper formation of intra-repeat disulfide bonds, which help to fix a conserved glycine residue at its proper position for Ca2+ coordination. Additionally, despite lacking inter repeat disulfide bonds, the interfaces between adjacent OmpA-TT3Rs are enhanced by both hydrophobic and conserved aromatic-proline interactions.« less

Protective efficacy of six immunogenic recombinant proteins of Vibrio anguillarum and evaluation them as vaccine candidate for flounder (Paralichthys olivaceus).

PubMed

Xing, Jing; Xu, Hongsen; Wang, Yang; Tang, Xiaoqian; Sheng, Xiuzhen; Zhan, Wenbin

2017-06-01

Vibrio anguillarum is a severe bacterium that causes terminal haemorrhagic septicaemia in freshwater and marine fish. Virulence-associated proteins play an important role in bacterial pathogenicity and could be applied for immunoprophylaxis. In this study, six antigenic proteins from V. anguillarum were selected and the immune protective efficacy of their recombinant proteins was investigated. VirA, CheR, FlaC, OmpK, OmpR and Hsp33 were recombinantly produced and the reactions of recombinant proteins to flounder-anti-V. anguillarum antibodies (fV-ab) were detected, respectively. Then the recombinant proteins were injected to fish, after immunization, the percentages of surface membrane immunoglobulin-positive (sIg+) cell in lymphocytes, total antibodies, antibodies against V. anguillarum, antibodies against recombinant proteins and relative percent survival (RPS) were analyzed, respectively. The results showed that all the recombinant proteins could react to fV-ab, proliferate sIg + cells in lymphocytes and induce production of total antibodies, specific antibodies against V. anguillarum or the recombinant proteins; the RPS of rVirA, rCheR, rFlaC, rOmpK, rOmpR and rHsp33 against V. anguillarum was 70.27%, 27.03%, 16.22%, 62.16%, 45.95% and 81.08%, respectively. The results revealed that rHsp33, rVirA and rOmpK have good protections against V. anguillarum and could be vaccine candidates against V. anguillarum. Copyright © 2017 Elsevier Ltd. All rights reserved.

Regulation of Toll-like receptor 2 interaction with Ecgp96 controls Escherichia coli K1 invasion of brain endothelial cells

PubMed Central

Krishnan, Subramanian; Chen, Shuang; Turcatel, Gianluca; Arditi, Moshe; Prasadarao, Nemani V.

2012-01-01

SUMMARY The interaction of outer membrane protein A (OmpA) with its receptor, Ecgp96 (a homologue of Hsp90β) is critical for the pathogenesis of E. coli K1 meningitis. Since Hsp90 chaperones Toll-like receptors (TLRs), we examined the role of TLRs in E. coli K1 infection. Herein, we show that newborn TLR2−/− mice are resistant to E. coli K1 meningitis, while TLR4−/− mice succumb to infection sooner. In vitro, OmpA+ E. coli infection selectively upregulates Ecgp96 and TLR2 in human brain microvascular endothelial cells (HBMEC), whereas OmpA− E. coli upregulates TLR4 in these cells. Furthermore, infection with OmpA+ E. coli causes Ecgp96 and TLR2 translocate to the plasma membrane of HBMEC as a complex. Immunoprecipitation studies of the plasma membrane fractions from infected HBMEC reveal that the C-termini of Ecgp96 and TLR2 are critical for OmpA+ E. coli invasion. Knockdown of TLR2 using siRNA results in inefficient membrane translocation of Ecgp96 and significantly reduces invasion. In addition, the interaction of Ecgp96 and TLR2 induces a bipartite signal, one from Ecgp96 through PKC-α while the other from TLR2 through MyD88, ERK1/2 and NF-κB. This bipartite signal ultimately culminates in the efficient production of NO, which in turn promotes E. coli K1 invasion of HBMEC. PMID:22963587

Regulation of Toll-like receptor 2 interaction with Ecgp96 controls Escherichia coli K1 invasion of brain endothelial cells.

PubMed

Krishnan, Subramanian; Chen, Shuang; Turcatel, Gianluca; Arditi, Moshe; Prasadarao, Nemani V

2013-01-01

The interaction of outer membrane protein A (OmpA) with its receptor, Ecgp96 (a homologue of Hsp90β), is critical for the pathogenesis of Escherichia coli K1 meningitis. Since Hsp90 chaperones Toll-like receptors (TLRs), we examined the role of TLRs in E. coli K1 infection. Herein, we show that newborn TLR2(-/-) mice are resistant to E. coli K1 meningitis, while TLR4(-/-) mice succumb to infection sooner. In vitro, OmpA+ E. coli infection selectively upregulates Ecgp96 and TLR2 in human brain microvascular endothelial cells (HBMEC), whereas OmpA- E. coli upregulates TLR4 in these cells. Furthermore, infection with OmpA+ E. coli causes Ecgp96 and TLR2 translocate to the plasma membrane of HBMEC as a complex. Immunoprecipitation studies of the plasma membrane fractions from infected HBMEC reveal that the C termini of Ecgp96 and TLR2 are critical for OmpA+ E. coli invasion. Knockdown of TLR2 using siRNA results in inefficient membrane translocation of Ecgp96 and significantly reduces invasion. In addition, the interaction of Ecgp96 andTLR2 induces a bipartite signal, one from Ecgp96 through PKC-α while the other from TLR2 through MyD88, ERK1/2 and NF-κB. This bipartite signal ultimately culminates in the efficient production of NO, which in turn promotes E. coli K1 invasion of HBMEC. © 2012 Blackwell Publishing Ltd.

Molecular Evolution and Mosaicism of Leptospiral Outer Membrane Proteins Involves Horizontal DNA Transfer

PubMed Central

Haake, David A.; Suchard, Marc A.; Kelley, Melissa M.; Dundoo, Manjula; Alt, David P.; Zuerner, Richard L.

2004-01-01

Leptospires belong to a genus of parasitic bacterial spirochetes that have adapted to a broad range of mammalian hosts. Mechanisms of leptospiral molecular evolution were explored by sequence analysis of four genes shared by 38 strains belonging to the core group of pathogenic Leptospira species: L. interrogans, L. kirschneri, L. noguchii, L. borgpetersenii, L. santarosai, and L. weilii. The 16S rRNA and lipL32 genes were highly conserved, and the lipL41 and ompL1 genes were significantly more variable. Synonymous substitutions are distributed throughout the ompL1 gene, whereas nonsynonymous substitutions are clustered in four variable regions encoding surface loops. While phylogenetic trees for the 16S, lipL32, and lipL41 genes were relatively stable, 8 of 38 (20%) ompL1 sequences had mosaic compositions consistent with horizontal transfer of DNA between related bacterial species. A novel Bayesian multiple change point model was used to identify the most likely sites of recombination and to determine the phylogenetic relatedness of the segments of the mosaic ompL1 genes. Segments of the mosaic ompL1 genes encoding two of the surface-exposed loops were likely acquired by horizontal transfer from a peregrine allele of unknown ancestry. Identification of the most likely sites of recombination with the Bayesian multiple change point model, an approach which has not previously been applied to prokaryotic gene sequence analysis, serves as a model for future studies of recombination in molecular evolution of genes. PMID:15090524

Application of the One-Minute Preceptor Technique by Novice Teachers in the Gross Anatomy Laboratory

ERIC Educational Resources Information Center

Chan, Lap Ki; Yang, Jian; Irby, David M.

2015-01-01

The one-minute preceptor (OMP) was originally developed in the ambulatory care setting as a time-efficient teaching technique for learner-centered clinical training. There are also possible advantages of using the OMP in the gross anatomy laboratory. However, in a previous study it was found that providing training to experienced gross anatomy…

Subcellular fractions of Brucella ovis distinctively induce the production of interleukin-2, interleukin-4, and interferon-γ in mice

PubMed Central

2005-01-01

Abstract The aim of this study was to evaluate the effect of 3 Brucella ovis subcellular protein fractions: Outer membrane (OMP), inner membrane (IMP), and cytoplasm (CP), on cellular immune response by in vitro production of interleukin (IL)-2, IL-4, and interferon (IFN)-γ. Each fraction was inoculated 3 times into Balb/c mice, primary cultures of mice spleen cells were done, and these were then stimulated with the fractions. Culture supernatants were collected at 24, 48, 72, 96, and 120 h postinoculation. Cytokine concentration was measured by Duoset-enzyme-linked immunosorbent assay (ELISA). The OMP fraction induced highest cellular immune response of 1000 pg/mL of IL-2 at 24 h, which decreased to < 100 pg/mL by 96 h. The IL-2 response for the IMP fraction was low at 24 h, but exceeded that of the OMP fraction at 72, 96, and 120 h. The CP showed a poor IL response. Regarding the IFN-γ production, OMP and IMP induced a high response at 120 h. These results open the possibility for the use of B. ovis outer and inner membrane proteins as a subcellular vaccine. PMID:15745223

Constitutive expression of the maltoporin LamB in the absence of OmpR damages the cell envelope.

PubMed

Reimann, Sylvia A; Wolfe, Alan J

2011-02-01

Cells experience multiple environmental stimuli simultaneously. To survive, they must respond accordingly. Unfortunately, the proper response to one stress easily could make the cell more susceptible to a second coexistent stress. To deal with such a problem, a cell must possess a mechanism that balances the need to respond simultaneously to both stresses. Our recent studies of ompR malT(Con) double mutants show that elevated expression of LamB, the outer membrane porin responsible for maltose uptake, causes cell death when the osmoregulator OmpR is disabled. To obtain insight into the nature of the death experienced by ompR malT(Con) mutants, we described the death process. On the basis of microscopic and biochemical approaches, we conclude that death results from a loss of membrane integrity. On the basis of an unbiased genome-wide search for suppressor mutations, we conclude that this loss of membrane integrity results from a LamB-induced envelope stress that the cells do not sufficiently perceive and thus do not adequately accommodate. Finally, we conclude that this envelope stress involves an imbalance in the lipopolysaccharide/porin composition of the outer membrane and an increased requirement for inorganic phosphate.

OMPS SDR Calibration and Validation

NASA Astrophysics Data System (ADS)

Sen, B.; Done, J.; Buss, R.; Jaross, G. R.; Kelly, T. J.

2009-12-01

The Ozone Mapper and Profiler Suite (OMPS) is scheduled to be launched on the NPOESS Preparatory Project (NPP) platform in early 2011. The OMPS will continue monitoring ozone from space, using three instruments, namely the Total Column Mapper (heritage: TOMS), the Nadir Profiler (heritage: SBUV) and the Limb Profiler (heritage: SOLSE/LORE). The Total Column Mapper (TC) sensor images the Earth through a slit, nadir-cell horizontally spaced at 49.5 km cross-track with an along-track reporting interval of 50 km. The total field of view (FOV) cross-track is 110 degree to provide daily global coverage. The TC sensor, a grating spectrometer, provides 0.45 nm spectral sampling across the wavelength range of 300-380 nm. The calibration stability, which is essential to enable long-term ozone monitoring, is maintained by periodic observations of the Sun, using a diffuser to redirect the solar irradiance into the sensor. We describe the plans to calibrate the TC sensor and validate the radiance data (TC Sensor Data Record or TC SDR) after launch. We discuss the measurements planned during the Intensive Cal/Val (ICV) phase of NPP mission, the data analysis methodology and results from the analysis of OMPS calibration measurements.

Entry and intracellular replication of Escherichia coli K1 in macrophages require expression of outer membrane protein A.

PubMed

Sukumaran, Sunil K; Shimada, Hiroyuki; Prasadarao, Nemani V

2003-10-01

Interactions between Escherichia coli K1, which causes meningitis in neonates, and macrophages have not been explored well. In this study we found that E. coli K1 was able to enter, survive, and replicate intracellularly in both murine and human macrophage cell lines, as well as in monocytes and macrophages of newborn rats. In addition, we demonstrated that OmpA (+) E. coli also enters and replicates in human peripheral blood monocytes in vitro. Outer membrane protein A (OmpA) expression on E. coli contributes to binding to macrophages, phagocytosis, and survival within macrophages. Opsonization with either complement proteins or antibody is not required for uptake and survival of the bacteria within the macrophages. Transmission electron microscopy and immunocytochemistry studies with the infected macrophages indicated that OmpA(+) E. coli multiplies enormously in a single phagosome and bursts the cell. Internalization of OmpA(+) E. coli by RAW 264.7 cells occurred by both actin- and microtubule-dependent processes, which are independent of RGD-mediated integrin receptors. Internalization and intracellular survival within phagocytic cells thus may play an important role in the development of bacteremia, which is crucial for E. coli crossing of the blood-brain barrier.

Nuclear matter parameters and optical model analysis of proton elastic scattering on the doubly magic nucleus 40Ca

NASA Astrophysics Data System (ADS)

Khalaf, A. M.; Khalifa, M. M.; Solieman, A. H. M.; Comsan, M. N. H.

2018-01-01

Owing to its doubly magic nature having equal numbers of protons and neutrons, the 40Ca nuclear scattering can be successfully described by the optical model that assumes a spherical nuclear potential. Therefore, optical model analysis was employed to calculate the elastic scattering cross section for p +40Ca interaction at energies from 9 to 22 MeV as well as the polarization at energies from 10 to 18.2 MeV. New optical model parameters (OMPs) were proposed based on the best fitting to experimental data. It is found that the best fit OMPs depend on the energy by smooth relationships. The results were compared with other OMPs sets regarding their chi square values (χ2). The obtained OMP's set was used to calculate the volume integral of the potentials and the root mean square (rms) value of nuclear matter radius of 40Ca. In addition, 40Ca bulk nuclear matter properties were discussed utilizing both the obtained rms radius and the Thomas-Fermi rms radius calculated using spherical Hartree-Fock formalism employing Skyrme type nucleon-nucleon force. The nuclear scattering SCAT2000 FORTRAN code was used for the optical model analysis.

A comprehensive analysis of the Omp85/TpsB protein superfamily structural diversity, taxonomic occurrence, and evolution

PubMed Central

Heinz, Eva; Lithgow, Trevor

2014-01-01

Members of the Omp85/TpsB protein superfamily are ubiquitously distributed in Gram-negative bacteria, and function in protein translocation (e.g., FhaC) or the assembly of outer membrane proteins (e.g., BamA). Several recent findings are suggestive of a further level of variation in the superfamily, including the identification of the novel membrane protein assembly factor TamA and protein translocase PlpD. To investigate the diversity and the causal evolutionary events, we undertook a comprehensive comparative sequence analysis of the Omp85/TpsB proteins. A total of 10 protein subfamilies were apparent, distinguished in their domain structure and sequence signatures. In addition to the proteins FhaC, BamA, and TamA, for which structural and functional information is available, are families of proteins with so far undescribed domain architectures linked to the Omp85 β-barrel domain. This study brings a classification structure to a dynamic protein superfamily of high interest given its essential function for Gram-negative bacteria as well as its diverse domain architecture, and we discuss several scenarios of putative functions of these so far undescribed proteins. PMID:25101071

Fate of leaf litter deposits and impacts on oxygen availability in bank filtration column studies.

PubMed

Bayarsaikhan, Uranchimeg; Filter, Josefine; Gernert, Ulrich; Jekel, Martin; Ruhl, Aki Sebastian

2018-07-01

Degradation of particulate organic carbon (POC) such as leaf litter might deplete dissolved oxygen within the upper layers of bank filtration, an efficient and robust barrier for pathogens and for various organic micro-pollutants (OMP) in water supply systems worldwide. The degradation of OMP during bank filtration depends on the redox conditions. The present study aimed at identifying the impacts and fates of different local leaves on the oxygen consumption and the possible biological degradation of indicator OMP. Oxygen concentrations initially decreased within the columns from around 8 mg/L in the influent to low concentrations indicating extensive consumption within a short travel distance. Still a substantial oxygen consumption was observed after 250 days. OMP concentrations were not significantly affected by the microbial processes. A layer of calcium carbonate crystallites was observed on the POC layer. Some leaf fragments appeared to be persistant towards degradation and the carbon content relative to nitrogen and sulfur contents decreased within 250 days. The results demonstrate that trees at bank filtration sites might have a strong long-term impact on the subsurface redox conditions. Copyright © 2018 Elsevier Inc. All rights reserved.

Acetyl phosphate and the phosphorylation of OmpR are involved in the regulation of the cell division rate in Escherichia coli.

PubMed

Prüss, B M

1998-09-01

Carbon sources that can be converted to acetate were added to the growth medium of Escherichia coli wild-type cells. Cells responded with an increased cell division rate. The addition of acetate also caused a decreased synthesis of flagella. Mutants in phosphotransacetylase, which are incapable of synthesizing acetyl phosphate, and mutants in the osmoregulator OmpR divided at a lower rate than did wild-type cells. The mutants did not increase their cell division rate upon the addition of serine, as observed for wild-type cells. These data are consistent with the idea that the previously described effect of serine upon the cell division rate is mediated by acetyl phosphate and phosphorylation of OmpR.

Metal Accumulation and Vanadium-Induced Multidrug Resistance by Environmental Isolates of Escherichia hermannii and Enterobacter cloacae

PubMed Central

Hernández, Alicia; Mellado, Rafael P.; Martínez, José L.

1998-01-01

Contaminated soils from an oil refinery were screened for the presence of microorganisms capable of accumulating either nickel, vanadium, or both metals. Three strains of bacteria that belonged to the family Enterobacteriaceae were selected. Two of them were Escherichia hermannii strains, and outer membrane profile (OMP) analysis showed that they were similar to a strain of clinical origin; the other one was an Enterobacter cloacae strain that differed from clinical isolates. The selected bacteria accumulated both nickel and vanadium. Growth in the presence of vanadium induced multidrug resistance phenotypes in E. hermannii and E. cloacae. Incubation with this metal changed the OMP profile of E. hermannii but did not produce variations in the expression of the major OMPs of E. cloacae. PMID:9797283

Electrostatic Interactions between OmpG Nanopore and Analyte Protein Surface Can Distinguish between Glycosylated Isoforms.

PubMed

Fahie, Monifa A; Chen, Min

2015-08-13

The flexible loops decorating the entrance of OmpG nanopore move dynamically during ionic current recording. The gating caused by these flexible loops changes when a target protein is bound. The gating is characterized by parameters including frequency, duration, and open-pore current, and these features combine to reveal the identity of a specific analyte protein. Here, we show that OmpG nanopore equipped with a biotin ligand can distinguish glycosylated and deglycosylated isoforms of avidin by their differences in surface charge. Our studies demonstrate that the direct interaction between the nanopore and analyte surface, induced by the electrostatic attraction between the two molecules, is essential for protein isoform detection. Our technique is remarkably sensitive to the analyte surface, which may provide a useful tool for glycoprotein profiling.

Comparative Proteomics of Tandem Mass Spectrometry Analyses for Bacterial Strains Identification and Differentiation

DTIC Science & Technology

2012-02-01

risk, bio -terrorism utility, Homeland Security, agricultural monitoring, quality of foodstuffs, environmental monitoring, and biological warfare agents...CAL19717 Putative surface antigen CAL21872 Putative sigma 54 modulation protein NP_395233 Plasminogen activator protease precursor CAL19882 OMP...S. (2005). Chemical and biological weapons : current concepts for future defenses. Johns Hopkins APL Tech. Digest, 26, 321-333. Dworzanski, J.P

Effects of Training Experienced Teachers in the Use of the One-Minute Preceptor Technique in the Gross Anatomy Laboratory

ERIC Educational Resources Information Center

Chan, Lap Ki; Sharma, Neel

2014-01-01

The one-minute preceptor (OMP) is a time-efficient, learner-centered teaching method used in a busy ambulatory care setting. This project evaluated the effects of training experienced anatomy teachers in the use of the OMP in the gross anatomy laboratory on students' perceived learning. Second-year medical students from a five-year,…

OMPS Limb Profiler: Extending SAGE and CALIPSO Stratospheric Aerosol Records

NASA Astrophysics Data System (ADS)

Taha, G.; Bhartia, P. K.; Chen, Z.; Xu, P.; Loughman, R. P.; Jaross, G.

2017-12-01

The OMPS LP instrument is designed to provide high vertical resolution ozone and aerosol profiles from measurements of the scattered solar radiation in the 290-1000 nm spectral range. It collected its first Earth limb measurement in January 10, 2012, and continues to provide daily global measurements of ozone and aerosol profiles from the cloud top up to 60 km and 40 km respectively. The relatively high vertical and spatial sampling allow detection and tracking periodic events when aerosol particles are injected into the stratosphere, such as volcanic eruptions or meteor explosions. OMPS LP can extend the long-term records of stratospheric aerosol at high vertical resolution produced by variety of sensors, such as SAGEII, GOMOS, OSIRIS and CALIPSO. Most of these instruments ceased to operate or well beyond their designed lifetime. After an absence of over a decade, SAGE III/ISS was launched earlier this year and expected to resume the high quality aerosol data record. OMPS LP is also schedule to fly on JPSS-2 and 3. In this study we will examine the suitability of using LP profiles to continue the stratospheric aerosol records beyond SAGE, OSIRIS, and CALIPSO. We will compare OMPS LP released V1.0 aerosol extinction measurements to OSIRIS and CALIPSO. Initial results shows good agreement with OSIRIS measurements to within 20%, with larger bias in the southern hemisphere. To test the effect of the assumed aerosol size model (ASD) and phase function, we compare measurements taken at similar location and time with different viewing geometry. Comparison of ascending and descending aerosol extinction daily zonal means at high latitudes shows systematic bias that is well correlated with the solar scattering angle, indicating ASD uncertainties up to 30%. In addition, results showing latitudinal, and temporal variability of stratospheric aerosol extinction and optical depth for the three instruments will also be presented and compared. We will also present OMPS LP aerosol observations of the dispersal of volcanic aerosols in the stratosphere following the eruptions of Kelut and Calbuco in 2014 and 2015 respectively.

Negative Effect of Proton-pump Inhibitors (PPIs) on Helicobacter pylori Growth, Morphology, and Urease Test and Recovery after PPI Removal--An In vitro Study.

PubMed

Saniee, Parastoo; Shahreza, Somayeh; Siavoshi, Farideh

2016-04-01

Proton-pump inhibitor (PPI) consumption does lead to false-negative results of Helicobacter pylori diagnostic tests such as biopsy culture and rapid urease test (RUT). Helicobacter pylori isolates from 112 dyspeptic patients with (56.5%) or without (43.5%) PPI consumption were recruited for examining the negative effects of omeprazole (OMP), lansoprazole (LPZ), and pantoprazole (PAN) on H. pylori viability, morphology, and urease, in vitro. The effect of a sublethal concentration of OMP on bacterial features and their recovery after removal of OMP was also assessed. Of 112 culture-positive gastric biopsies, 87.5% were RUT positive and 12.5% RUT negative. There was a significant correlation between negative RUT results and PPI consumption (p < .05). OMP (minimum inhibitory concentration, MIC 32 μg/mL) and LPZ (MIC 8 μg/mL) inhibited the growth of 78.6% of H. pylori isolates. OMP and LPZ inhibited urease of 90.3% of isolates between 0 and 40 minutes and 54.4% between 20 and 40 minutes, respectively. PAN did not inhibit H. pylori growth and urease. Three 3-day (9 days) consecutive subcultures of H. pylori on brucella blood agar (BBA) supplemented with OMP resulted in reduced bacterial viability (1+), compared with control (4+), change of spiral morphology to coccoid, and reduction in pink color intensity in urea agar. Bacterial growth (1+), morphology, and urease test did not improve after the first 3-day and second 3-day (6 days) subcultures on BBA. However, relative recovery occurred after the third 3-day (9 days) subculture and complete recovery was observed after the fourth 3-day (12 days) subculture, as confluent growth (4+), 100% spiral cells, and strong urease test. Proton-pump Inhibitors exert transient negative effects on H. pylori viability, morphology, and urease test. Accordingly, cessation of PPI consumption at least 12 days before endoscopy could help avoiding false-negative results of H. pylori diagnostic tests. © 2015 John Wiley & Sons Ltd.

Reconciling uncertainty of costs and outcomes with the need for access to orphan medicinal products: a comparative study of managed entry agreements across seven European countries.

PubMed

Morel, Thomas; Arickx, Francis; Befrits, Gustaf; Siviero, Paolo; van der Meijden, Caroline; Xoxi, Entela; Simoens, Steven

2013-12-24

National payers across Europe have been increasingly looking into innovative reimbursement approaches - called managed entry agreements (MEAs) - to balance the need to provide rapid access to potentially beneficial orphan medicinal products (OMPs) with the requirements to circumscribe uncertainty, obtain best value for money or to ensure affordability. This study aimed to identify, describe and classify MEAs applied to OMPs by national payers and to analyse their practice in Europe. To identify and describe MEAs, national health technology assessments and reimbursement decisions on OMPs across seven European countries were reviewed and their main characteristics extracted. To fill data gaps and validate the accuracy of the extraction, collaboration was sought from national payers. To classify MEAs, a bespoke taxonomy was implemented. Identified MEAs were analysed and compared by focusing on five key themes, namely by describing the MEAs in relation to: drug targets and therapeutic classes, geographical spread, type of MEA applied, declared rationale for setting-up of MEAs, and evolution over time. 42 MEAs for 26 OMPs, implemented between 2006 and 2012 and representing a variety of MEA designs, were identified. Italy was the country with the highest number of schemes (n=15), followed by the Netherlands (n=10), England and Wales (n=8), Sweden (n=5) and Belgium (n=4). No MEA was identified for France and Germany due to data unavailability. Antineoplastic agents were the primary targets of MEAs. 55% of the identified MEAs were performance-based risk-sharing arrangements; the other 45% were financial-based. Nine of these 26 OMPs were subject to MEAs in two or three different countries, resulting in 24 MEAs. 60% of identified MEAs focused on conditions whose prevalence is less than 1 per 10,000. This study confirmed that a variety of MEAs were increasingly used by European payers to manage aspects of uncertainty associated with the introduction of OMPs in the healthcare system, and which may be of a clinical, utilisation, or budgetary nature. It remains unclear whether differences in the use of MEAs reflect differences in how 'uncertainty' and 'value' are perceived across healthcare systems.

Development of RP UPLC-TOF/MS, stability indicating method for omeprazole and its related substances by applying two level factorial design; and identification and synthesis of non-pharmacopoeial impurities.

PubMed

Jadhav, Sushant Bhimrao; Kumar, C Kiran; Bandichhor, Rakeshwar; Bhosale, P N

2016-01-25

A new UPLC-TOF/MS compatible, reverse phase-stability indicating method was developed for determination of Omeprazole (OMP) and its related substances in pharmaceutical dosage forms by implementing Design of Experiment (DoE) i.e. two level full factorial Design (2(3)+3 center points=11 experiments) to understand the Critical Method Parameters (CMP) and its relation with Critical Method Attribute (CMA); to ensure robustness of the method. The separation of eleven specified impurities including conversion product of OMP related compound F (13) and G (14) i.e. Impurity-I (1), OMP related compound-I (11) and OMP 4-chloro analog (12) was achieved in a single method on Acquity BEH shield RP18 100 × 2.1 mm, 1.7 μm column, with inlet filter (0.2 μm) using gradient elution and detector wavelength at 305 nm and validated in accordance with ICH guidelines and found to be accurate, precise, reproducible, robust and specific. The drug was found to degrade extensively in heat, humidity and acidic conditions and forms unknown degradation products during stability studies. The same method was used for LC-MS analysis to identify m/z and fragmentation of maximum unknown impurities (Non-Pharmacopoeial) i.e. Impurity-I (1), Impurity-III (3), Impurity-V (5) and Impurity-VIII (9) formed during stability studies. Based on the results, degradation pathway for the drug has been proposed and synthesis of identified impurities i.e. impurities (Impurity-I (1), Impurity-III (3), Impurity-V (5) and Impurity-VIII (9)) are discussed in detail to ensure in-depth understanding of OMP and its related impurities and optimum performance during lifetime of the product. Copyright © 2015. Published by Elsevier B.V.

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

PubMed Central

Van Loock, Marnix; Verminnen, Kristel; Messmer, Trudy O; Volckaert, Guido; Goddeeris, Bruno M; Vanrompay, Daisy

2005-01-01

Background Laboratory diagnosis of Chlamydophila psittaci, an important turkey respiratory pathogen, is difficult. To facilitate the diagnosis, a nested PCR-enzyme immunoassay (PCR-EIA) was developed to detect the Cp. psittaci outer membrane protein A (ompA) gene in pharyngeal swabs. Methods The fluorescein-biotin labelled PCR products were immobilized on streptavidin-coated microtiter plates and detected with anti-fluorescein peroxidase conjugate and a colorimetric substrate. An internal inhibition control was included to rule out the presence of inhibitors of DNA amplification. The diagnostic value of the ompA nested PCR-EIA in comparison to cell culture and a 16S-rRNA based nested PCR was assessed in pharyngeal turkey swabs from 10 different farms experiencing respiratory disease. Results The sensitivity of the nested PCR-EIA was established at 0.1 infection forming units (IFU). Specificity was 100%. The ompA nested PCR-EIA was more sensitive than the 16S-rRNA based nested PCR and isolation, revealing 105 out of 200 (52.5%) positives against 13 and 74 for the latter two tests, respectively. Twenty-nine (23.8%) out of 122 ompA PCR-EIA negatives showed the presence of inhibitors of DNA amplification, although 27 of them became positive after diluting (1/10) the specimens in PCR buffer or after phenol-chloroform extraction and subsequent ethanol precipitation. Conclusion The present study stresses the need for an internal control to confirm PCR true-negatives and demonstrates the high prevalence of chlamydiosis in Belgian turkeys and its potential zoonotic risk. The ompA nested PCR-EIA described here is a rapid, highly sensitive and specific diagnostic assay and will help to facilitate the diagnosis of Cp. psittaci infections in both poultry and man. PMID:16185353

Aerosol Correction for Improving OMPS/LP Ozone Retrieval

NASA Technical Reports Server (NTRS)

Chen, Zhong; Bhartia, Pawan K.; Loughman, Robert

2015-01-01

The Ozone Mapping and Profiler Suite Limb Profiler (OMPS-LP) on board the Suomi National Polar-orbiting Partnership (SNPP) satellite was launched on Oct. 28, 2011. Limb profilers measures the radiance scattered from the Earth's atmospheric in limb viewing mode from 290 to 1000 nm and infer ozone profiles from tropopause to 60 km. The recently released OMPS-LP Version 2 data product contains the first publicly released ozone profiles retrievals, and these are now available for the entire OMPS mission, which extends from April, 2012. The Version 2 data product retrievals incorporate several important improvements to the algorithm. One of the primary changes is to turn off the aerosol retrieval module. The aerosol profiles retrieved inside the ozone code was not helping the ozone retrieval and was adding noise and other artifacts. Aerosols including polar stratospheric cloud (PSC) and polar mesospheric clouds (PMC) have a detectable effect on OMPS-LP data. Our results show that ignoring the aerosol contribution would produce an ozone density bias of up to 10 percent in the region of maximum aerosol extinction. Therefore, aerosol correction is needed to improve the quality of the retrieved ozone concentration profile. We provide Aerosol Scattering Index (ASI) for detecting aerosols-PMC-PSC, defined as ln(Im-Ic) normalized at 45km, where Im is the measured radiance and Ic is the calculated radiance assuming no aerosols. Since ASI varies with wavelengths, latitude and altitude, we can start by assuming no aerosol profiles in calculating the ASIs and then use the aerosol profile to see if it significantly reduces the residuals. We also discuss the effect of aerosol size distribution on the ozone profile retrieval process. Finally, we present an aerosol-PMC-PSC correction scheme.

Logarithmic phase Escherichia coli K1 efficiently avoids serum killing by promoting C4bp-mediated C3b and C4b degradation

PubMed Central

Wooster, David G; Maruvada, Ravi; Blom, Anna M; Prasadarao, Nemani V

2006-01-01

Meningitis caused by Escherichia coli K1 is a serious illness in neonates with neurological sequelae in up to 50% of survivors. A high degree of bacteremia is required for E. coli K1 to cross the blood–brain barrier, which suggests that the bacterium must evade the host defence mechanisms and survive in the bloodstream. We previously showed that outer membrane protein A (OmpA) of E. coli binds C4b-binding protein (C4bp), an inhibitor of complement activation via the classical pathway. Nevertheless, the exact mechanism by which E. coli K1 survives in serum remains elusive. Here, we demonstrate that log phase (LP) OmpA+E. coli K1 avoids serum bactericidal activity more effectively than postexponential phase bacteria. OmpA–E. coli cannot survive in serum grown to either phase. The increased serum resistance of LP OmpA+E. coli is the result of increased binding of C4bp, with a concomitant decrease in the deposition of C3b and the downstream complement proteins responsible for the formation of the membrane attack complex. C4bp bound to E. coli K1 acts as a cofactor to factor I in the cleavage of both C3b and C4b, which shuts down the ensuing complement cascade. Accordingly, a peptide corresponding to the complement control protein domain 3 of C4bp sequence, was able to compete with C4bp binding to OmpA and cause increased deposition of C3b. Thus, binding of C4bp appears to be responsible for survival of E. coli K1 in human serum. PMID:16556262

Spermatogenetic inhibition in men taking a combination of oral medroxyprogesterone acetate and percutaneous testosterone as a male contraceptive method.

PubMed

Soufir, J-C; Meduri, G; Ziyyat, A

2011-07-01

We previously demonstrated in a small pilot study that oral medroxyprogesterone acetate and percutaneous testosterone (OMP/PT) induce reversible spermatogenesis suppression. The aims of this study were to determine the rate of spermatogenetic inhibition and recovery and to obtain preliminary data on efficacy for a larger population under OMP/PT. A total of 35 healthy men with normal spermiograms requesting male hormonal contraception were treated with OMP (20 mg/day) and PT (50-125 mg/day) for periods up to 18 months. Couples were included in a contraceptive efficacy phase after a value of ≤1 million/ml spermatozoa was reached between 1 and 3 months of treatment. Sperm counts decreased by 47% at 1 month, reaching 90% at 2 months and 98-100% between 4 and 8 months. At 3 months, 80% of men had ≤1 million/ml spermatozoa. Follicle-stimulating hormone and luteinizing hormone decreased to 35% of pretreatment levels after 1 month of treatment and to 75-80% at 2 and 6 months, respectively. Plasma testosterone and estradiol levels were in the eugonadal range at 3, 6, 9 and 12 months of treatment. Dihydrotestosterone concentrations were 2-4 times higher than pretreatment values. The rate of spermatogenetic recovery was rapid (73 ± 29.5 days). During the efficacy phase (211 months for 25 couples), one pregnancy attributable to poor compliance of the male partner was observed. OMP/PT efficiently inhibits spermatogenesis in 80% of men, maintains testosterone at physiological levels and avoids the need for parenteral administration, which is poorly accepted by French men. These results justify larger studies to define a more adequate dosage of OMP/PT and to confirm its efficacy and safety.

Transcription of TP0126, Treponema pallidum Putative OmpW Homolog, Is Regulated by the Length of a Homopolymeric Guanosine Repeat

PubMed Central

Brandt, Stephanie L.; Ke, Wujian; Reid, Tara B.; Molini, Barbara J.; Iverson-Cabral, Stefanie; Ciccarese, Giulia; Drago, Francesco; Lukehart, Sheila A.; Centurion-Lara, Arturo

2015-01-01

An effective mechanism for introduction of phenotypic diversity within a bacterial population exploits changes in the length of repetitive DNA elements located within gene promoters. This phenomenon, known as phase variation, causes rapid activation or silencing of gene expression and fosters bacterial adaptation to new or changing environments. Phase variation often occurs in surface-exposed proteins, and in Treponema pallidum subsp. pallidum, the syphilis agent, it was reported to affect transcription of three putative outer membrane protein (OMP)-encoding genes. When the T. pallidum subsp. pallidum Nichols strain genome was initially annotated, the TP0126 open reading frame was predicted to include a poly(G) tract and did not appear to have a predicted signal sequence that might suggest the possibility of its being an OMP. Here we show that the initial annotation was incorrect, that this poly(G) is instead located within the TP0126 promoter, and that it varies in length in vivo during experimental syphilis. Additionally, we show that TP0126 transcription is affected by changes in the poly(G) length consistent with regulation by phase variation. In silico analysis of the TP0126 open reading frame based on the experimentally identified transcriptional start site shortens this hypothetical protein by 69 amino acids, reveals a predicted cleavable signal peptide, and suggests structural homology with the OmpW family of porins. Circular dichroism of recombinant TP0126 supports structural homology to OmpW. Together with the evidence that TP0126 is fully conserved among T. pallidum subspecies and strains, these data suggest an important role for TP0126 in T. pallidum biology and syphilis pathogenesis. PMID:25802057

The Atypical Response Regulator Protein ChxR Has Structural Characteristics and Dimer Interface Interactions That Are Unique within the OmpR/PhoB Subfamily

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hickey, John M.; Lovell, Scott; Battaile, Kevin P.

2013-05-29

Typically as a result of phosphorylation, OmpR/PhoB response regulators form homodimers through a receiver domain as an integral step in transcriptional activation. Phosphorylation stabilizes the ionic and hydrophobic interactions between monomers. Recent studies have shown that some response regulators retain functional activity in the absence of phosphorylation and are termed atypical response regulators. The two currently available receiver domain structures of atypical response regulators are very similar to their phospho-accepting homologs, and their propensity to form homodimers is generally retained. An atypical response regulator, ChxR, from Chlamydia trachomatis, was previously reported to form homodimers; however, the residues critical to thismore » interaction have not been elucidated. We hypothesize that the intra- and intermolecular interactions involved in forming a transcriptionally competent ChxR are distinct from the canonical phosphorylation (activation) paradigm in the OmpR/PhoB response regulator subfamily. To test this hypothesis, structural and functional studies were performed on the receiver domain of ChxR. Two crystal structures of the receiver domain were solved with the recently developed method using triiodo compound I3C. These structures revealed many characteristics unique to OmpR/PhoB subfamily members: typical or atypical. Included was the absence of two {alpha}-helices present in all other OmpR/PhoB response regulators. Functional studies on various dimer interface residues demonstrated that ChxR forms relatively stable homodimers through hydrophobic interactions, and disruption of these can be accomplished with the introduction of a charged residue within the dimer interface. A gel shift study with monomeric ChxR supports that dimerization through the receiver domain is critical for interaction with DNA.« less

New insights into the effects of support matrix on the removal of organic micro-pollutants and the microbial community in constructed wetlands.

PubMed

Zhang, Liang; Lyu, Tao; Ramírez Vargas, Carlos Andrés; Arias, Carlos A; Carvalho, Pedro N; Brix, Hans

2018-09-01

Constructed wetlands (CWs) are an eco-friendly and cost-effective technology to remove organic micro-pollutants (OMPs) from wastewater. The support matrix is an important component in CWs as it has a primary role in the growth and development of plants and microbes. However, the roles of the support matrix in CWs in removing OMPs have not been systematically studied. Therefore, in this study, six common materials (sand, zeolite, blast iron slag, petcoke, polonite and crushed autoclaved aerated concrete (CAAC)) as support matrixes were firstly investigated by batch tests to explore their adsorption capacities to selected OMPs (ibuprofen, iohexol, tebuconazole and imazalil). Results showed that the adsorption capacities of the materials were low (at the level of μg/g) compared to well-known sorbents (at the level of mg/g), such as activated carbon and carbon nanotubes. Columns packed with the six materials, respectively, were then built up to study the effects of different materials on microbial community. In the medium-term study (66 days), the removal of four OMPs in all the columns increased by 2-58% from day 25 to day 66, and was mainly attributed to microbial degradation. Furthermore, Community-level physiological profiling (CLPP) analysis indicates that material presence shaped the microbial community metabolic function not only in the interstitial water but also in the biofilm. Overall, all the findings demonstrate that although the adsorption capacities of the common materials are low, they may be a driver to improve the removal of OMPs by altering microbial community function in CWs. Copyright © 2018 Elsevier Ltd. All rights reserved.

Interaction of E. coli outer-membrane protein A with sugars on the receptors of the brain microvascular endothelial cells.

PubMed

Datta, Deepshikha; Vaidehi, Nagarajan; Floriano, Wely B; Kim, Kwang S; Prasadarao, Nemani V; Goddard, William A

2003-02-01

Esherichia coli, the most common gram-negative bacteria, can penetrate the brain microvascular endothelial cells (BMECs) during the neonatal period to cause meningitis with significant morbidity and mortality. Experimental studies have shown that outer-membrane protein A (OmpA) of E. coli plays a key role in the initial steps of the invasion process by binding to specific sugar moieties present on the glycoproteins of BMEC. These experiments also show that polymers of chitobiose (GlcNAcbeta1-4GlcNAc) block the invasion, while epitopes substituted with the L-fucosyl group do not. We used HierDock computational technique that consists of a hierarchy of coarse grain docking method with molecular dynamics (MD) to predict the binding sites and energies of interactions of GlcNAcbeta1-4GlcNAc and other sugars with OmpA. The results suggest two important binding sites for the interaction of carbohydrate epitopes of BMEC glycoproteins to OmpA. We identify one site as the binding pocket for chitobiose (GlcNAcbeta1-4GlcNAc) in OmpA, while the second region (including loops 1 and 2) may be important for recognition of specific sugars. We find that the site involving loops 1 and 2 has relative binding energies that correlate well with experimental observations. This theoretical study elucidates the interaction sites of chitobiose with OmpA and the binding site predictions made in this article are testable either by mutation studies or invasion assays. These results can be further extended in suggesting possible peptide antagonists and drug design for therapeutic strategies. Copyright 2002 Wiley-Liss, Inc.

Application of Fourier transform infrared spectroscopy and chemometrics for differentiation of Salmonella enterica serovar Enteritidis phage types.

PubMed

Preisner, Ornella; Guiomar, Raquel; Machado, Jorge; Menezes, José Cardoso; Lopes, João Almeida

2010-06-01

Fourier transform infrared (FT-IR) spectroscopy and chemometric techniques were used to discriminate five closely related Salmonella enterica serotype Enteritidis phage types, phage type 1 (PT1), PT1b, PT4b, PT6, and PT6a. Intact cells and outer membrane protein (OMP) extracts from bacterial cell membranes were subjected to FT-IR analysis in transmittance mode. Spectra were collected over a wavenumber range from 4,000 to 600 cm(-1). Partial least-squares discriminant analysis (PLS-DA) was used to develop calibration models based on preprocessed FT-IR spectra. The analysis based on OMP extracts provided greater separation between the Salmonella Enteritidis PT1-PT1b, PT4b, and PT6-PT6a groups than the intact cell analysis. When these three phage type groups were considered, the method based on OMP extract FT-IR spectra was 100% accurate. Moreover, complementary local models that considered only the PT1-PT1b and PT6-PT6a groups were developed, and the level of discrimination increased. PT1 and PT1b isolates were differentiated successfully with the local model using the entire OMP extract spectrum (98.3% correct predictions), whereas the accuracy of discrimination between PT6 and PT6a isolates was 86.0%. Isolates belonging to different phage types (PT19, PT20, and PT21) were used with the model to test its robustness. For the first time it was demonstrated that FT-IR analysis of OMP extracts can be used for construction of robust models that allow fast and accurate discrimination of different Salmonella Enteritidis phage types.

Immunogenicity and protective efficacy of recombinant Haemophilus parasuis SH0165 putative outer membrane proteins.

PubMed

Fu, Shulin; Zhang, Minmin; Xu, Juan; Ou, Jiwen; Wang, Yan; Liu, Huazhen; Liu, Jinlin; Chen, Huanchun; Bei, Weicheng

2013-01-02

Haemophilus parasuis (H. parasuis), the causative agent of swine polyserositis, polyarthritis, and meningitis, is one of the most important bacterial diseases of pigs worldwide. Little vaccines currently exist that have a significant effect on infections with all pathogenic serovars of H. parasuis. H. parasuis putative outer membrane proteins (OMPs) are potentially essential components of more effective vaccines. Recently, the genomic sequence of H. parasuis serovar 5 strain SH0165 was completed in our laboratory, which allow us to target OMPs for the development of recombinant vaccines. In this study, we focused on 10 putative OMPs and all the putative OMPs were cloned, expressed and purified as HIS fusion proteins. Primary screening for immunoprotective potential was performed in mice challenged with an LD50 challenge. Out of these 10 OMPs three fusion proteins rGAPDH, rOapA, and rHPS-0675 were found to be protective in a mouse model of H. parasuis infection. We further evaluated the immune responses and protective efficacy of rGAPDH, rOapA, and rHPS-0675 in pig models. All three proteins elicited humoral antibody responses and conferred different levels of protection against challenge with a lethal dose of H. parasuis SH0165 in pig models. In addition, the antisera against the three individual proteins and the synergistic protein efficiently inhibited bacterial growth in a whole blood assay. The data demonstrated that the three proteins showed high value individually and the combination of rGAPDH, rOapA, and rHPS-0675 offered the best protection. Our results indicate that rGAPDH, rOapA, and rHPS-0675 induced protection against H. parasuis SH0165 infection, which may facilitate the development of a multi-component vaccine. Copyright © 2012 Elsevier Ltd. All rights reserved.

Identification of minimum carbohydrate moiety in N-glycosylation sites of brain endothelial cell glycoprotein 96 for interaction with Escherichia coli K1 outer membrane protein A.

PubMed

Krishnan, Subramanian; Prasadarao, Nemani V

2014-07-01

Bacterial meningitis is a serious central nervous system infection and Escherichia coli K1 (E. coli K1) is one of the leading etiological agents that cause meningitis in neonates. Outer membrane protein A (OmpA) of E. coli K1 is a major virulence factor in the pathogenesis of meningitis, and interacts with human brain microvascular endothelial cells (HBMEC) to cross the blood-brain barrier. Using site-directed mutagenesis, we demonstrate that two N-glycosylation sites (NG1 and NG2) in the extracellular domain of OmpA receptor, Ecgp96 are critical for bacterial binding to HBMEC. E. coli K1 invasion assays using CHO-Lec1 cells that express truncated N-glycans, and sequential digestion of HBMEC surface N-glycans using specific glycosidases showed that GlcNAc1-4GlcNAc epitopes are sufficient for OmpA interaction with HBMEC. Lack of NG1 and NG2 sites in Ecgp96 inhibits E. coli K1 OmpA induced F-actin polymerization, phosphorylation of protein kinase C-α, and disruption of transendothelial electrical resistance required for efficient invasion of E. coli K1 in HBMEC. Furthermore, the microvessels of cortex and hippocampus of the brain sections of E. coli K1 infected mice showed increased expression of glycosylated Ecgp96. Therefore, the interface of OmpA and GlcNAc1-4GlcNAc epitope interaction would be a target for preventative strategies against E. coli K1 meningitis. Copyright © 2014 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

Evaluation of antiulcer activity of indole-3-carbinol and/or omeprazole on aspirin-induced gastric ulcer in rats.

PubMed

El-Shinnawy, Nashwa A; Abd-Elmageid, Samira A; Alshailabi, Eda M A

2014-05-01

The present work is an attempt to elucidate the antiulcer activity of indole-3-carbinol (I3C), which is one of the anticarcinogenic phytochemicals found in the vegetables of Cruciferae family such as broccoli and cauliflower, alone or in combination with omeprazole (OMP), a proton pump inhibitor, to diminish the effects of induced acute gastric ulcer by aspirin (ASA) in male albino rats. A total of 48 adult male albino rats were used in the present study. Animals were divided into eight experimental groups (six animals each group). They were given different experimental inductions of ASA at a dose of 500 mg/kg/body weight, OMP at a dose of 20 mg/kg/body weight and I3C at a dose of 20 mg/kg/body weight either alone or in combination with each other orally for a duration of 7 days. Inner stomach features, ulcer index, pH activity, body weight, stomach weight, hematological investigations, serum total protein albumin and reduced glutathione activity were investigated in addition to the histological, histochemical and immunohistochemical stain of cyclooxygenase-2 to the stomach tissue of normal control, ulcerated and treated ulcerated rats. The results of this study revealed that oral administration of ASA to rats produced the expected characteristic mucosal lesions. OMP accelerated ulcer healing but the administration of I3C either alone or in combination with OMP to ASA-ulcerated rats produced a profound protection to the gastric mucosa from injury induced by ASA. Our results suggested that administration of antiulcer natural substances such as I3C in combination with the perused treatment such as OMP is a very important initiative in the development of new strategies in ulcer healing.

Molecular mechanisms of membrane impermeability in clinical isolates of Enterobacteriaceae exposed to imipenem selective pressure.

PubMed

Pavez, Monica; Vieira, Camila; de Araujo, Maria Rita; Cerda, Alvaro; de Almeida, Lara Mendes; Lincopan, Nilton; Mamizuka, Elsa Masae

2016-07-01

Intrinsic mechanisms leading to carbapenem-induced membrane impermeability and multidrug resistance are poorly understood in clinical isolates of Enterobacteriaceae. In this study, molecular behaviours during the establishment of membrane impermeability in members of the Enterobacteriaceae family under imipenem selective pressure were investigated. Clinical isolates (n = 22) exhibiting susceptibility to multiple antibiotics or characterised as extended-spectrum β-lactamase (ESBL)- or AmpC-producers were submitted to progressive passages on Mueller-Hinton agar plates containing subclinical concentrations of imipenem [0.5 × the minimum inhibitory concentration (MIC)]. Changes in outer membrane permeability were evaluated by determination of antimicrobial MICs, sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), and gene expression analysis related to membrane permeability (i.e. omp35-like, omp36-like and acrA) and regulatory mechanisms (i.e. marA and ompR) by quantitative reverse transcription PCR. Following imipenem induction, 73% of isolates showed increased carbapenem MICs by ≥2 doubling dilutions. At an early stage of treatment, imipenem modulated the expression of porins and efflux pump genes, represented by a reduction of 78% in omp36-like and a two-fold increase in acrA expression. Transcriptional factors marA and ompR were also affected by imipenem induction, increasing mRNA expression by 14- and 4-fold, respectively. High marA expression levels were associated with higher values of acrA expression. These results suggest that imipenem is an important factor in the development of an adaptive response to carbapenems by regulating key genes involved in the control of efflux pumps and porins, which could lead to a multidrug-resistant profile in clinical isolates, contributing to possible treatment failure. Copyright © 2016 Elsevier B.V. and International Society of Chemotherapy. All rights reserved.

Gene cloning and prokaryotic expression of recombinant outer membrane protein from Vibrio parahaemolyticus

NASA Astrophysics Data System (ADS)

Yuan, Ye; Wang, Xiuli; Guo, Sheping; Qiu, Xuemei

2011-06-01

Gram-negative Vibrio parahaemolyticus is a common pathogen in humans and marine animals. The outer membrane protein of bacteria plays an important role in the infection and pathogenicity to the host. Thus, the outer membrane proteins are an ideal target for vaccines. We amplified a complete outer membrane protein gene (ompW) from V. parahaemolyticus ATCC 17802. We then cloned and expressed the gene into Escherichia coli BL21 (DE3) cells. The gene coded for a protein that was 42.78 kDa. We purified the protein using Ni-NTA affinity chromatography and Anti-His antibody Western blotting, respectively. Our results provide a basis for future application of the OmpW protein as a vaccine candidate against infection by V. parahaemolyticus. In addition, the purified OmpW protein can be used for further functional and structural studies.

The structure of the β-barrel assembly machinery complex

DOE PAGES

Bakelar, Jeremy; Buchanan, Susan K.; Noinaj, Nicholas

2016-01-08

β-Barrel outer membrane proteins (OMPs) are found in the outer membranes of Gram-negative bacteria and are essential for nutrient import, signaling, and adhesion. A 200-kilodalton five-component complex called the β-barrel assembly machinery (BAM) complex has been implicated in the biogenesis of OMPs. In this paper, we report the structure of the BAM complex from Escherichia coli, revealing that binding of BamCDE modulates the conformation of BamA, the central component, which may serve to regulate the BAM complex. The periplasmic domain of BamA was in a closed state that prevents access to the barrel lumen, which indicates substrate OMPs may notmore » be threaded through the barrel during biogenesis. Finally and further, conformational shifts in the barrel domain lead to opening of the exit pore and rearrangement at the lateral gate.« less

The structure of the β-barrel assembly machinery complex

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bakelar, Jeremy; Buchanan, Susan K.; Noinaj, Nicholas

β-Barrel outer membrane proteins (OMPs) are found in the outer membranes of Gram-negative bacteria and are essential for nutrient import, signaling, and adhesion. A 200-kilodalton five-component complex called the β-barrel assembly machinery (BAM) complex has been implicated in the biogenesis of OMPs. In this paper, we report the structure of the BAM complex from Escherichia coli, revealing that binding of BamCDE modulates the conformation of BamA, the central component, which may serve to regulate the BAM complex. The periplasmic domain of BamA was in a closed state that prevents access to the barrel lumen, which indicates substrate OMPs may notmore » be threaded through the barrel during biogenesis. Finally and further, conformational shifts in the barrel domain lead to opening of the exit pore and rearrangement at the lateral gate.« less

The in silico screening and X-ray structure analysis of the inhibitor complex of Plasmodium falciparum orotidine 5'-monophosphate decarboxylase.

PubMed

Takashima, Yasuhide; Mizohata, Eiichi; Krungkrai, Sudaratana R; Fukunishi, Yoshifumi; Kinoshita, Takayoshi; Sakata, Tsuneaki; Matsumura, Hiroyoshi; Krungkrai, Jerapan; Horii, Toshihiro; Inoue, Tsuyoshi

2012-08-01

Orotidine 5'-monophosphate decarboxylase from Plasmodium falciparum (PfOMPDC) catalyses the final step in the de novo synthesis of uridine 5'-monophosphate (UMP) from orotidine 5'-monophosphate (OMP). A defective PfOMPDC enzyme is lethal to the parasite. Novel in silico screening methods were performed to select 14 inhibitors against PfOMPDC, with a high hit rate of 9%. X-ray structure analysis of PfOMPDC in complex with one of the inhibitors, 4-(2-hydroxy-4-methoxyphenyl)-4-oxobutanoic acid, was carried out to at 2.1 Å resolution. The crystal structure revealed that the inhibitor molecule occupied a part of the active site that overlaps with the phosphate-binding region in the OMP- or UMP-bound complexes. Space occupied by the pyrimidine and ribose rings of OMP or UMP was not occupied by this inhibitor. The carboxyl group of the inhibitor caused a dramatic movement of the L1 and L2 loops that play a role in the recognition of the substrate and product molecules. Combining part of the inhibitor molecule with moieties of the pyrimidine and ribose rings of OMP and UMP represents a suitable avenue for further development of anti-malarial drugs.

The fitness costs and trade-off shapes associated with the exclusion of nine antibiotics by OmpF porin channels.

PubMed

Phan, Katherine; Ferenci, Thomas

2017-06-01

The trade-off relationship between antibiotic exclusion and nutrient access across the Gram-negative outer membrane is determined by structural constraints in porin channels. The precise nutritional cost of exclusion is unknown for different antibiotics, as are the shapes of the nutrition-susceptibility trade-off. Using a library of 10 engineered isogenic Escherichia coli strains with structural modifications of OmpF porin expressed at a constant level, susceptibilities were measured for nine antibiotics and the nutritional fitness costs estimated by competitions in chemostats. Different antibiotics exhibited a remarkably varied range of geometries in the nutrition-susceptibility trade-off, including convex, concave and sigmoidal trade-off shapes. The trade-off patterns predict the possibility of adaptations in contributing to antibiotic resistance; exclusion of amoxicillin or trimethoprim in ompF mutants can occur with little loss of fitness whereas kanamycin and streptomycin exclusion has a high cost. Some individual OmpF changes even allow positive correlations (trade-ups), resulting in increased fitness and decreased susceptibility specifically to cephalexin or ciprofloxacin. The surprising plasticity of the nutrition-exclusion relationship means that there are no generalisable rules that apply to decreasing susceptibility for all antibiotics. The protein changes are exquisitely specific in determining nutritional fitness and adaptive outcomes in a structural constraint trade-off.

Constitutive Expression of the Maltoporin LamB in the Absence of OmpR Damages the Cell Envelope▿ †

PubMed Central

Reimann, Sylvia A.; Wolfe, Alan J.

2011-01-01

Cells experience multiple environmental stimuli simultaneously. To survive, they must respond accordingly. Unfortunately, the proper response to one stress easily could make the cell more susceptible to a second coexistent stress. To deal with such a problem, a cell must possess a mechanism that balances the need to respond simultaneously to both stresses. Our recent studies of ompR malT(Con) double mutants show that elevated expression of LamB, the outer membrane porin responsible for maltose uptake, causes cell death when the osmoregulator OmpR is disabled. To obtain insight into the nature of the death experienced by ompR malT(Con) mutants, we described the death process. On the basis of microscopic and biochemical approaches, we conclude that death results from a loss of membrane integrity. On the basis of an unbiased genome-wide search for suppressor mutations, we conclude that this loss of membrane integrity results from a LamB-induced envelope stress that the cells do not sufficiently perceive and thus do not adequately accommodate. Finally, we conclude that this envelope stress involves an imbalance in the lipopolysaccharide/porin composition of the outer membrane and an increased requirement for inorganic phosphate. PMID:21131484

Putative Inv Is Essential for Basolateral Invasion of Caco-2 Cells and Acts Synergistically with OmpA To Affect In Vitro and In Vivo Virulence of Cronobacter sakazakii ATCC 29544

PubMed Central

Chandrapala, Dilini; Kim, Kyumson; Choi, Younho; Senevirathne, Amal; Kang, Dong-Hyun; Ryu, Sangryeol

2014-01-01

Cronobacter sakazakii is an opportunistic pathogen that causes neonatal meningitis and necrotizing enterocolitis. Its interaction with intestinal epithelium is important in the pathogenesis of enteric infections. In this study, we investigated the involvement of the inv gene in the virulence of C. sakazakii ATCC 29544 in vitro and in vivo. Sequence analysis of C. sakazakii ATCC 29544 inv revealed that it is different from other C. sakazakii isolates. In various cell culture models, an Δinv deletion mutant showed significantly lowered invasion efficiency, which was restored upon genetic complementation. Studying invasion potentials using tight-junction-disrupted Caco-2 cells suggested that the inv gene product mediates basolateral invasion of C. sakazakii ATCC 29544. In addition, comparison of invasion potentials of double mutant (ΔompA Δinv) and single mutants (ΔompA and Δinv) provided evidence for an additive effect of the two putative outer membrane proteins. Finally, the importance of inv and the additive effect of putative Inv and OmpA were also proven in an in vivo rat pup model. This report is the first to demonstrate two proteins working synergistically in vitro, as well as in vivo in C. sakazakii pathogenesis. PMID:24549330

Putative Inv is essential for basolateral invasion of Caco-2 cells and acts synergistically with OmpA to affect in vitro and in vivo virulence of Cronobacter sakazakii ATCC 29544.

PubMed

Chandrapala, Dilini; Kim, Kyumson; Choi, Younho; Senevirathne, Amal; Kang, Dong-Hyun; Ryu, Sangryeol; Kim, Kwang-Pyo

2014-05-01

Cronobacter sakazakii is an opportunistic pathogen that causes neonatal meningitis and necrotizing enterocolitis. Its interaction with intestinal epithelium is important in the pathogenesis of enteric infections. In this study, we investigated the involvement of the inv gene in the virulence of C. sakazakii ATCC 29544 in vitro and in vivo. Sequence analysis of C. sakazakii ATCC 29544 inv revealed that it is different from other C. sakazakii isolates. In various cell culture models, an Δinv deletion mutant showed significantly lowered invasion efficiency, which was restored upon genetic complementation. Studying invasion potentials using tight-junction-disrupted Caco-2 cells suggested that the inv gene product mediates basolateral invasion of C. sakazakii ATCC 29544. In addition, comparison of invasion potentials of double mutant (ΔompA Δinv) and single mutants (ΔompA and Δinv) provided evidence for an additive effect of the two putative outer membrane proteins. Finally, the importance of inv and the additive effect of putative Inv and OmpA were also proven in an in vivo rat pup model. This report is the first to demonstrate two proteins working synergistically in vitro, as well as in vivo in C. sakazakii pathogenesis.

Development and antigenic characterization of three recombinant proteins with potential for Glässer's disease prevention.

PubMed

Li, Miao; Li, Chunling; Song, Shuai; Kang, Huahua; Yang, Dongxia; Li, Guoqing

2016-04-27

Haemophilus parasuis is the causative agent of Glässer's disease, which causes high morbidity and mortality in piglets, leading to severe economic losses. The lack of a commercial vaccine against a broad spectrum of strains has limited the disease control. H. parasuis outer membrane proteins (OMPs) are potentially essential components for vaccine formulation. In this study, seven putative OMPs were selected from the annotated H. parasuis serovar 5 genome; they were predicted by bioinformatics and annotated as potential virulence-related factors. These proteins were cloned, expressed, and purified as His-tagged proteins. Antigenicity of the candidate proteins was assessed using Western blotting with convalescent sera against H. parasuis serovar 5. The immunogenicity of the seven OMPs was assessed in a guinea pig model. Except VacJ, all the other six recombinant proteins elicited a detectable antibody response. Antisera against four of the selected proteins effectively killed the bacteria in vitro. Three proteins (Omp26, VacJ, and HAPS_0742) were found to confer significant protection against challenge with a lethal dose of H. parasuis in a guinea pig model. The results suggest that these three proteins have a strong potential to be vaccine candidates against Glässer's disease. Copyright © 2016 Elsevier Ltd. All rights reserved.

Improving Fab' fragment retention in an autonucleolytic Escherichia coli strain by swapping periplasmic nuclease translocation signal from OmpA to DsbA.

PubMed

Schofield, Desmond M; Sirka, Ernestas; Keshavarz-Moore, Eli; Ward, John M; Nesbeth, Darren N

2017-12-01

To reduce unwanted Fab' leakage from an autonucleolytic Escherichia coli strain, which co-expresses OmpA-signalled Staphylococcal nuclease and Fab' fragment in the periplasm, by substituting in Serratial nuclease and the DsbA periplasm translocation signal as alternatives. We attempted to genetically fuse a nuclease from Serratia marcescens to the OmpA signal peptide but plasmid construction failed, possibly due to toxicity of the resultant nuclease. Combining Serratial nuclease to the DsbA signal peptide was successful. The strain co-expressing this nuclease and periplasmic Fab' grew in complex media and exhibited nuclease activity detectable by DNAse agar plate but its growth in defined medium was retarded. Fab' coexpression with Staphylococcal nuclease fused to the DsbA signal peptide resulted in cells exhibiting nuclease activity and growth in defined medium. In cultivation to high cell density in a 5 l bioreactor, DsbA-fused Staphylococcal nuclease co-expression coincided with reduced Fab' leakage relative to the original autonucleolytic Fab' strain with OmpA-fused staphylococcal nuclease. We successfully rescued Fab' leakage back to acceptable levels and established a basis for future investigation of the linkage between periplasmic nuclease expression and leakage of co-expressed periplasmic Fab' fragment to the surrounding growth media.

Removal of micropollutants with coarse-ground activated carbon for enhanced separation with hydrocyclone classifiers.

PubMed

Otto, N; Platz, S; Fink, T; Wutscherk, M; Menzel, U

2016-01-01

One key technology to eliminate organic micropollutants (OMP) from wastewater effluent is adsorption using powdered activated carbon (PAC). To avoid a discharge of highly loaded PAC particles into natural water bodies a separation stage has to be implemented. Commonly large settling tanks and flocculation filters with the application of coagulants and flocculation aids are used. In this study, a multi-hydrocyclone classifier with a downstream cloth filter has been investigated on a pilot plant as a space-saving alternative with no need for a dosing of chemical additives. To improve the separation, a coarser ground PAC type was compared to a standard PAC type with regard to elimination results of OMP as well as separation performance. With a PAC dosing rate of 20 mg/l an average of 64.7 wt% of the standard PAC and 79.5 wt% of the coarse-ground PAC could be separated in the hydrocyclone classifier. A total average separation efficiency of 93-97 wt% could be reached with a combination of both hydrocyclone classifier and cloth filter. Nonetheless, the OMP elimination of the coarse-ground PAC was not sufficient enough to compete with the standard PAC. Further research and development is necessary to find applicable coarse-grained PAC types with adequate OMP elimination capabilities.

Deciphering the roles of outer membrane protein A extracellular loops in the pathogenesis of Escherichia coli K1 meningitis.

PubMed

Mittal, Rahul; Krishnan, Subramanian; Gonzalez-Gomez, Ignacio; Prasadarao, Nemani V

2011-01-21

Outer membrane protein A (OmpA) has been implicated as an important virulence factor in several gram-negative bacterial infections such as Escherichia coli K1, a leading cause of neonatal meningitis associated with significant mortality and morbidity. In this study, we generated E. coli K1 mutants that express OmpA in which three or four amino acids from various extracellular loops were changed to alanines, and we examined their ability to survive in several immune cells. We observed that loop regions 1 and 2 play an important role in the survival of E. coli K1 inside neutrophils and dendritic cells, and loop regions 1 and 3 are needed for survival in macrophages. Concomitantly, E. coli K1 mutants expressing loop 1 and 2 mutations were unable to cause meningitis in a newborn mouse model. Of note, mutations in loop 4 of OmpA enhance the severity of the pathogenesis by allowing the pathogen to survive better in circulation and to produce high bacteremia levels. These results demonstrate, for the first time, the roles played by different regions of extracellular loops of OmpA of E. coli K1 in the pathogenesis of meningitis and may help in designing effective preventive strategies against this deadly disease.

The OMPS Limb Profiler instrument

NASA Astrophysics Data System (ADS)

Rault, D. F.; Xu, P.

2011-12-01

The Ozone Mapping and Profiler Suite (OMPS) will continue the monitoring of the global distribution of the Earth's middle atmosphere ozone and aerosol. OMPS is composed of three instruments, namely the Total Column Mapper (heritage: TOMS, OMI), the Nadir Profiler (heritage: SBUV) and the Limb Profiler (heritage: SOLSE/LORE, OSIRIS, SCIAMACHY, SAGE III). The ultimate goal of the mission is to better understand and quantify the rate of stratospheric ozone recovery. OMPS is scheduled to be launched on the NPOESS Preparatory Project (NPP) platform in October 2011. The focus of the paper will be on the Limb Profiler (LP) instrument. The LP instrument will measure the Earth's limb radiance, from which ozone profile will be retrieved from the upper tropopause uo to 60km. End-to-end studies of the sensor and retrieval algorithm indicate the following expected performance for ozone: accuracy of 5% or better from the tropopause up to 50 km, precision of about 3-5% from 18 to 50 km, and vertical resolution of 1.5-2 km with vertical sampling of 1 km and along-track horizontal sampling of 1 deg latitude. The paper will describe the mission, discuss the retrieval algorithm, and summarize the expected performance. If available, the paper will also present early on-orbit data.

First isolation of Rickettsia monacensis from a patient in South Korea.

PubMed

Kim, Yeon-Sook; Choi, Yeon-Joo; Lee, Kyung-Min; Ahn, Kyu-Joong; Kim, Heung-Chul; Klein, Terry; Jiang, Ju; Richards, Allen; Park, Kyung-Hee; Jang, Won-Jong

2017-07-01

A Rickettsia sp. was isolated from the blood of a patient with an acute febrile illness using the shell vial technique; the isolate was named CN45Kr and was identified by molecular assay as Rickettsia monacensis, which was first recognized as a pathogen in Spain. Sequencing analysis showed that the gltA sequence of the isolate was identical to that of Rickettsia sp. IRS3. The ompA-5mp fragment sequence showed 100% identity to those of R. monacensis and Rickettsia sp. In56 and ompA-3pA In56 and 100% identity to that of Rickettsia sp. IRS3. The ompB sequence was found to have 99.9% similarity to that of R. monacensis IrR/Munich. This study confirms the pathogenicity of this agent and provides additional information about its geographic distribution. © 2017 The Societies and John Wiley & Sons Australia, Ltd.

New Stratospheric Dust Belt Due to the Chelyabinsk Bolide

NASA Technical Reports Server (NTRS)

Gorkavyi, Nick; Rault, D. F.; Newman, P. A.; da Silva, A. M.; Dudorov, A. E.

2013-01-01

The Ozone Mapping Profiler Suite (OMPS) Limb Profiler (LP) on the recently launched NASA/NOAA NPP/ Suomi satellite detected aerosol excess in the midstratosphere (25-45 km altitude) between 50degN and 70degN latitudes. OMPS/LP observations trace this aerosol plume to the meteor that struck near Chelyabinsk, Russia on 15 February 2013. This new dust layer, located above the Junge aerosol layer, has persisted over at least a 3 month period. Material collected on the ground following the bolide explosion showed that the meteor was mostly composed of olivine and pyroxenes. Simulations using Lagrangian and Eulerian atmospheric models trace the plume back to Chelyabinsk and confirm that the plume altitude was at altitudes between 25 and 45 km. The models also confirm the plume circumpolar longitudinal spreading observed by OMPS/LP, with propagation speeds up to 85 m/s.

Combination of granular activated carbon adsorption and deep-bed filtration as a single advanced wastewater treatment step for organic micropollutant and phosphorus removal.

PubMed

Altmann, Johannes; Rehfeld, Daniel; Träder, Kai; Sperlich, Alexander; Jekel, Martin

2016-04-01

Adsorption onto granular activated carbon (GAC) is an established technology in water and advanced wastewater treatment for the removal of organic substances from the liquid phase. Besides adsorption, the removal of particulate matter by filtration and biodegradation of organic substances in GAC contactors has frequently been reported. The application of GAC as both adsorbent for organic micropollutant (OMP) removal and filter medium for solids retention in tertiary wastewater filtration represents an energy- and space saving option, but has rarely been considered because high dissolved organic carbon (DOC) and suspended solids concentrations in the influent of the GAC adsorber put a significant burden on this integrated treatment step and might result in frequent backwashing and unsatisfactory filtration efficiency. This pilot-scale study investigates the combination of GAC adsorption and deep-bed filtration with coagulation as a single advanced treatment step for simultaneous removal of OMPs and phosphorus from secondary effluent. GAC was assessed as upper filter layer in dual-media downflow filtration and as mono-media upflow filter with regard to filtration performance and OMP removal. Both filtration concepts effectively removed suspended solids and phosphorus, achieving effluent concentrations of 0.1 mg/L TP and 1 mg/L TSS, respectively. Analysis of grain size distribution and head loss within the filter bed showed that considerable head loss occurred in the topmost filter layer in downflow filtration, indicating that most particles do not penetrate deeply into the filter bed. Upflow filtration exhibited substantially lower head loss and effective utilization of the whole filter bed. Well-adsorbing OMPs (e.g. benzotriazole, carbamazepine) were removed by >80% up to throughputs of 8000-10,000 bed volumes (BV), whereas weakly to medium adsorbing OMPs (e.g. primidone, sulfamethoxazole) showed removals <80% at <5,000 BV. In addition, breakthrough behavior was also determined for gabapentin, an anticonvulsant drug recently detected in drinking water resources for which suitable removal technologies are still largely unknown. Gabapentin showed poor adsorptive removal, resulting in rapid concentration increases. Whereas previous studies classified gabapentin as not readily biodegradable, sustained removal was observed after prolonged operation and points at biological elimination of gabapentin within the GAC filter. The application of GAC as filter medium was compared to direct addition of powdered activated carbon (PAC) to deep-bed filtration as a direct process alternative. Both options yielded comparable OMP removals for most compounds at similar carbon usage rates, but GAC achieved considerably higher removals for biodegradable OMPs. Based on the results, the application of GAC in combination with coagulation/filtration represents a promising alternative to powdered activated carbon and ozone for advanced wastewater treatment. Copyright © 2016 Elsevier Ltd. All rights reserved.

The Use of OMPS Near Real Time Products in Volcanic Cloud Risk Mitigation and Smoke/Dust Air Quality Assessments

NASA Astrophysics Data System (ADS)

Seftor, C. J.; Krotkov, N. A.; McPeters, R. D.; Li, J. Y.; Durbin, P. B.

2015-12-01

Near real time (NRT) SO2 and aerosol index (AI) imagery from Aura's Ozone Monitoring Instrument (OMI) has proven invaluable in mitigating the risk posed to air traffic by SO2 and ash clouds from volcanic eruptions. The OMI products, generated as part of NASA's Land, Atmosphere Near real-time Capability for EOS (LANCE) NRT system and available through LANCE and both NOAA's NESDIS and ESA's Support to Aviation Control Service (SACS) portals, are used to monitor the current location of volcanic clouds and to provide input into Volcanic Ash (VA) advisory forecasts. NRT products have recently been developed using data from the Ozone Mapping and Profiler Suite onboard the Suomi NPP platform; they are currently being made available through the SACS portal and will shortly be incorporated into the LANCE NRT system. We will show examples of the use of OMPS NRT SO2 and AI imagery to monitor recent volcanic eruption events. We will also demonstrate the usefulness of OMPS AI imagery to detect and track dust storms and smoke from fires, and how this information can be used to forecast their impact on air quality in areas far removed from their source. Finally, we will show SO2 and AI imagery generated from our OMPS Direct Broadcast data to highlight the capability of our real time system.

Volcanic Ash and SO2 Monitoring Using Suomi NPP Direct Broadcast OMPS Data

NASA Astrophysics Data System (ADS)

Seftor, C. J.; Krotkov, N. A.; McPeters, R. D.; Li, J. Y.; Brentzel, K. W.; Habib, S.; Hassinen, S.; Heinrichs, T. A.; Schneider, D. J.

2014-12-01

NASA's Suomi NPP Ozone Science Team, in conjunction with Goddard Space Flight Center's (GSFC's) Direct Readout Laboratory, developed the capability of processing, in real-time, direct readout (DR) data from the Ozone Mapping and Profiler Suite (OMPS) to perform SO2 and Aerosol Index (AI) retrievals. The ability to retrieve this information from real-time processing of DR data was originally developed for the Ozone Monitoring Instrument (OMI) onboard the Aura spacecraft and is used by Volcano Observatories and Volcanic Ash Advisory Centers (VAACs) charged with mapping ash clouds from volcanic eruptions and providing predictions/forecasts about where the ash will go. The resulting real-time SO2 and AI products help to mitigate the effects of eruptions such as the ones from Eyjafjallajokull in Iceland and Puyehue-Cordón Caulle in Chile, which cause massive disruptions to airline flight routes for weeks as airlines struggle to avoid ash clouds that could cause engine failure, deeply pitted windshields impossible to see through, and other catastrophic events. We will discuss the implementation of real-time processing of OMPS DR data by both the Geographic Information Network of Alaska (GINA) and the Finnish Meteorological Institute (FMI), which provide real-time coverage over some of the most congested airspace and over many of the most active volcanoes in the world, and show examples of OMPS DR processing results from recent volcanic eruptions.

The detection of antigenic determinants of Acinetobacter baumannii.

PubMed

Tawfik, Dina M; Ahmad, Tarek A; Sheweita, Salah A; Haroun, Medhat; El-Sayed, Laila H

2017-06-01

Acinetobacter baumannii continues to pose a threat to burdened patients in ICUs all around the world. Lately, infection control techniques are not sufficient to curb A. baumannii's progression and chemotherapeutics are losing their potency against it. Thus, immunization became a key player in providing an ideal solution to the dilemma. None of the vaccines under investigation have reached the market and the search for a tailored vaccine remains a challenge. The notion of unravelling the bacterial antigens to design a novel epitope-based vaccine proved its merits. In this work, the propitious polysaccharide and protein antigenic determinants of A. baumannii were mapped by mimicking the infection. The immune response was evaluated by western blot, ELISA, and cellular proliferation assay techniques. The screening showed that OMPs induced the most eminent sustained IgG response. In addition, OMP gave the highest cellular proliferation and a fold increase in ELISA that reached up to 10-fold by week 6. Whilst, the LPS gave a rapid IgM response, that reached 5-fold and the response was visible from week 1 in the western blot. The OMPs had a more pronounced effect in eliciting a cellular immune response. The results elaborated the valuable role of using pure OMPs and detoxified LPS together; as a major cornerstone in designing an ideal vaccine against A. baumannii. Copyright © 2017 European Federation of Immunological Societies. Published by Elsevier B.V. All rights reserved.

The Analysis of Inter-calibration Between FY-3C/TOU, NPP/OMPS and Metop/GOME-2

NASA Astrophysics Data System (ADS)

Wang, H.; Hu, X.

2017-12-01

Total ozone unit (TOU), one of the main payloads on FY-3C satellite, is the instrument for daily global coverage of total ozone monitoring in China. It has been in-orbit for about four years since October 2013. However, its solar irradiance is not correct because all of three diffuser boards cannot work normally. Therefore, in-orbit inter-calibration of radiance and reflectance are studied for TOU measurement. A method is introduced for inter-calibration between FY-3C/TOU and NPP/OMPS, Metop-B/GOME-2. It includes orbit forecast, temporal concurrent, spatial collocation, geometrical alignement, uniform filtration, and spectral consistent. Then, it is used for TOU data of 3 years from 2014 to 2016. The slopes of radiance inter-calibration equations of 360 nm between TOU (y-axis) and NPP/OMPS (x-axis) decrease gradually from 1 to 0.96. The slopes of radiance inter-calibration equations of 360 nm between TOU (y-axis) and Metop-B/GOME2 (x-axis) increased gradually from 1.12 to 1.72, while the slopes between TOU and Metop-A/GOME2 varied within 2.1-2.3. Most relation coefficients (R2) of them are >0.8. The inter-calibration results, combining with the solar irradiance of OMPS/GOME-2, will be used for the attenuation analysis of TOU measurements.

What's the Evidence: A Review of the One-Minute Preceptor Model of Clinical Teaching and Implications for Teaching in the Emergency Department.

PubMed

Farrell, Susan E; Hopson, Laura R; Wolff, Margaret; Hemphill, Robin R; Santen, Sally A

2016-09-01

The 2012 Academic Emergency Medicine Consensus Conference, "Education Research in Emergency Medicine: Opportunities, Challenges, and Strategies for Success" noted that emergency medicine (EM) educators often rely on theory and tradition in molding their approaches to teaching and learning, and called on the EM education community to advance the teaching of our specialty through the performance and application of research in teaching and assessment methods, cognitive function, and the effects of education interventions. The purpose of this article is to review the research-based evidence for the effectiveness of the one-minute preceptor (OMP) teaching method, and to provide suggestions for its use in clinical teaching and learning in EM. This article reviews hypothesis-testing education research related to the use of the OMP as a pedagogical method applicable to clinical teaching. Evidence indicates that the OMP prompts the teaching of higher level concepts, facilitates the assessment of students' knowledge, and prompts the provision of feedback. Students indicate satisfaction with this method of clinical case-based discussion teaching. Advancing EM education will require that high quality education research results be translated into actual curricular, pedagogical, assessment, and professional development changes. The OMP is a pedagogical method that is applicable to teaching in the emergency department. Copyright © 2016 Elsevier Inc. All rights reserved.

Understanding the fate of organic micropollutants in sand and granular activated carbon biofiltration systems.

PubMed

Paredes, L; Fernandez-Fontaina, E; Lema, J M; Omil, F; Carballa, M

2016-05-01

In this study, sand and granular activated carbon (GAC) biofilters were comparatively assessed as post-treatment technologies of secondary effluents, including the fate of 18 organic micropollutants (OMPs). To determine the contribution of adsorption and biotransformation in OMP removal, four reactors were operated (two biofilters (with biological activity) and two filters (without biological activity)). In addition, the influence of empty bed contact time (EBCT), ranging from 0.012 to 3.2d, and type of secondary effluent (anaerobic and aerobic) were evaluated. Organic matter, ammonium and nitrate were removed in both biofilters, being their adsorption higher on GAC than on sand. According to the behaviour exhibited, OMPs were classified in three different categories: I) biotransformation and high adsorption on GAC and sand (galaxolide, tonalide, celestolide and triclosan), II) biotransformation, high adsorption on GAC but low or null adsorption on sand (ibuprofen, naproxen, fluoxetine, erythromycin, roxythromycim, sulfamethoxazole, trimethoprim, bisphenol A, estrone, 17β-estradiol and 17α-ethinylestradiol), and, III) only adsorption on GAC (carbamazepine, diazepam and diclofenac). No influence of EBCT (in the range tested) and type of secondary effluent was observed in GAC reactors, whereas saturation and kinetic limitation of biotransformation were observed in sand reactors. Taking into account that most of the organic micropollutants studied (around 60%) fell into category II, biotransformation is crucial for the elimination of OMPs in sand biofilters. Copyright © 2016 Elsevier B.V. All rights reserved.

Impacts of backwashing on granular activated carbon filters for advanced wastewater treatment.

PubMed

Frank, Joshua; Ruhl, Aki Sebastian; Jekel, Martin

2015-12-15

The use of granular activated carbon (GAC) in fixed bed filters is a promising option for the removal of organic micropollutants (OMP) from wastewater treatment plant effluents. Frequent backwashing of the filter bed is inevitable, but its effect on potential filter stratification is not well understood yet and thus has been evaluated in the present study for two commercial GAC products. Backwashing of GAC filters was simulated with 10 or 100 filter bed expansions of 20 or 100% at backwash velocities of 12 and 40 m/h, respectively. Five vertical fractions were extracted and revealed a vertical stratification according to grain sizes and material densities. Sieve analyses indicated increasing grain sizes towards the bottom for one GAC while grain sizes of the other GAC were more homogeneously distributed throughout the filter bed. The apparent densities of the top sections were significantly lower than that of the bottom sections of both products. Comparative long term fixed bed adsorption experiments with the top and bottom sections of the stratified GAC showed remarkable differences in breakthrough curves of dissolved organic carbon, UV light absorption at 254 nm wavelength (UVA254) and OMP. GAC from the upper section showed constantly better removal efficiencies than GAC from the bottom section, especially for weakly adsorbing OMP such as sulfamethoxazole. Furthermore correlations between UVA254 reductions and OMP removals were found. Copyright © 2015 Elsevier Ltd. All rights reserved.

A molecular dynamics study of the pores formed by Escherichia coli OmpF porin in a fully hydrated palmitoyloleoylphosphatidylcholine bilayer.

PubMed

Tieleman, D P; Berendsen, H J

1998-06-01

In this paper we study the properties of pores formed by OmpF porin from Escherichia coli, based on a molecular dynamics simulation of the OmpF trimer, 318 palmitoyl-oleoyl-phosphatidylethanolamine lipids, 27 Na+ ions, and 12,992 water molecules. After equilibration and a nanosecond production run, the OmpF trimer exhibits a C-alpha root mean square deviation from the crystal structure of 0.23 nm and a stable secondary structure. No evidence is found for large-scale motions of the L3 loop. We investigate the pore dimensions, conductance, and the properties of water inside the pore. This water forms a complicated pattern, even when averaged over 1 ns of simulation time. Around the pore constriction zone the water dipoles are highly structured in the plane of the membrane, oriented by the strong transversal electric field. In addition, there is a net orientation along the pore axis pointing from the extracellular to the intracellular side of the bilayer. The diffusion coefficients of water inside the pore are greatly reduced compared to bulk. We compare our results to results from model pores (Breed et al., 1996. Biophys. J. 70:1 643-1 661; Sansom et al. 1997. Biophys. J. 73:2404-241 5) and discuss implications for further theoretical work.

Confinement of a β-barrel protein in nanoperforated free-standing nanomembranes for ion transport.

PubMed

Puiggalí-Jou, Anna; Pérez-Madrigal, Maria M; Del Valle, Luis J; Armelin, Elaine; Casas, María T; Michaux, Catherine; Perpète, Eric A; Estrany, Francesc; Alemán, Carlos

2016-09-29

Bioinspired free-standing nanomembranes (FSNMs) for selective ion transport have been tailored by immobilizing the Omp2a β-barrel membrane protein inside nanoperforations created in flexible poly(lactic acid) (PLA) nanomembranes. Perforated PLA FSNMs have been prepared by spin-coating a 99 : 1 PLA : poly(vinyl alcohol) mixture, and through a phase segregation process nanofeatures with dimensions similar to the entire nanomembrane thickness (∼110 nm) were induced. These nanofeatures have subsequently been transformed into nanoperforations (diameter: ∼51 nm) by selective solvent etching. The protein confined inside the nanopores of PLA FSNMs preserves the β-barrel structure and organizes in ovoid aggregates. The transport properties of Na + , K + , and Ca 2+ across non-perforated PLA, nanoperforated PLA, and Omp2a-filled nanoperforated PLA have been monitored by measuring the nanomembrane resistance with electrochemical impedance spectroscopy (EIS). The incorporation of nanoperforations enhances the transport of ions across PLA nanomembranes, whereas the functionality of immobilized Omp2a is essential to exhibit effects similar to those observed in biological nanomembranes. Indeed, Omp2a-filled nanoperforated PLA nanomembranes exhibit stronger affinity towards Na + and Ca 2+ ions than towards K + . In summary, this work provides a novel bioinspired strategy to develop mechanically stable and flexible FSNMs with channels for ion transport, which are precisely located inside artificial nanoperforations, thus holding great potential for applications in biofiltration and biosensing.

Evaluation of a membrane bioreactor system as post-treatment in waste water treatment for better removal of micropollutants.

PubMed

Arriaga, Sonia; de Jonge, Nadieh; Nielsen, Marc Lund; Andersen, Henrik Rasmus; Borregaard, Vibeke; Jewel, Kevin; Ternes, Thomas A; Nielsen, Jeppe Lund

2016-12-15

Organic micropollutants (OMPs) such as pharmaceuticals are persistent pollutants that are only partially degraded in waste water treatment plants (WWTPs). In this study, a membrane bioreactor (MBR) system was used as a polishing step on a full-scale WWTP, and its ability to remove micropollutants was examined together with the development and stability of the microbial community. Two stages of operation were studied during a period of 9 months, one with (S1) and one without (S2) the addition of exogenous OMPs. Ibuprofen and naproxen had the highest degradation rates with values of 248 μg/g VSS ·h and 71 μg/g VSS ·h, whereas diclofenac was a more persistent OMP (7.28 μg/g VSS ·h). Mineralization of 14 C-labeled OMPs in batch kinetic experiments indicates that higher removal rates (∼0.8 ng/mg T SS ·h) with a short lag phase can be obtained when artificial addition of organic micropollutants was performed. Similar microbial populations dominated S1 and S2, despite the independent operations. Hydrogenophaga, Nitrospira, p55-a5, the actinobacterial Tetrasphaera, Propionicimonas, Fodinicola, and Candidatus Microthrix were the most abundant groups in the polishing MBR. Finally, potential microbial candidates for ibuprofen and naproxen degradation are proposed. Copyright © 2016 Elsevier Ltd. All rights reserved.

77 FR 27827 - Self-Regulatory Organizations; Chicago Mercantile Exchange Inc.; Notice of Filing and Immediate...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-05-11

... margin calculations for these portfolios and perform ``what if'' analyses. The portfolio margining... Change Relating to Adoption of Interest Rate Futures Contracts Portfolio Margining Program With Eris... adopt an interest rate futures contracts portfolio margining program with Eris Exchange, LLC (``Eris...

In vivo roles of BamA, BamB and BamD in the biogenesis of BamA, a core protein of the β-barrel assembly machine of Escherichia coli.

PubMed

Misra, Rajeev; Stikeleather, Ryan; Gabriele, Rebecca

2015-03-13

Assembly of the β-barrel outer membrane proteins (OMPs) is an essential cellular process in Gram-negative bacteria and in the mitochondria and chloroplasts of eukaryotes--two organelles of bacterial origin. Central to this process is the conserved β-barrel OMP that belongs to the Omp85 superfamily. In Escherichia coli, BamA is the core β-barrel OMP and, together with four outer membrane lipoproteins, BamBCDE, constitutes the β-barrel assembly machine (BAM). In this paper, we investigated the roles of BamD, an essential lipoprotein, and BamB in BamA biogenesis. Depletion of BamD caused impairment in BamA biogenesis and cessation of cell growth. These defects of BamD depletion were partly reversed by single-amino-acid substitutions mapping within the β-barrel domain of BamA. However, in the absence of BamB, the positive effects of the β-barrel substitutions on BamA biogenesis under BamD depletion conditions were nullified. By employing a BamA protein bearing one such substitution, F474L, it was demonstrated that the mutant BamA protein could not only assemble without BamD but also facilitate the assembly of wild-type BamA expressed in trans. Based on these data, we propose a model in which the Bam lipoproteins, which are localized to the outer membrane by the BAM-independent Lol pathway, aid in the creation of new BAM complexes by serving as outer membrane receptors and folding factors for nascent BamA molecules. The newly assembled BAM holocomplex then catalyzes the assembly of substrate OMPs and BamA. These in vivo findings are corroborated by recently published in vitro data. Copyright © 2014 Elsevier Ltd. All rights reserved.

Using MERRA-2 analysis fields to simulate limb scattered radiance profiles for inhomogeneous atmospheric lines of sight: Preparation for data assimilation of OMPS LP radiances through 2D single-scattering GSLS radiative transfer model development

NASA Astrophysics Data System (ADS)

Loughman, R. P.; Bhartia, P. K.; Moy, L.; Kramarova, N. A.; Wargan, K.

2016-12-01

Many remote sensing techniques used to monitor the Earth's upper atmosphere fall into the broad category of "limb viewing" (LV) measurements, which includes any method for which the line of sight (LOS) fails to intersect the surface. Occultation, limb emission and limb scattering (LS) measurements are all LV methods that offer strong sensitivity to changes in the atmosphere near the tangent point of the LOS, due to the enhanced geometric path through the tangent layer (where the concentration also typically peaks, for most atmospheric species). But many of the retrieval algorithms used to interpret LV measurements assume that the atmosphere consists of "spherical shells", in which the atmospheric properties vary only with altitude (creating a 1D atmosphere). This assumption simplifies the analysis, but at the possible price of misinterpreting measurements made in the real atmosphere. In this presentation, we focus on the problem of LOS inhomogeneity for LS measurements made by the OMPS Limb Profiler (LP) instrument during the 2015 ozone hole period. The GSLS radiative transfer model (RTM) used in the default OMPS LP algorithms assumes a spherical-shell atmosphere defined at levels spaced 1 km apart, with extinction coefficients assumed to vary linearly with height between levels. Several recent improvements enable an updated single-scattering version of the GSLS RTM to ingest 3D MERRA-2 analysis fields (including temperature, pressure, and ozone concentration) when creating the model atmosphere, by introducing flexible altitude grids, flexible atmospheric specification along the LOS, and improved treatment of the radiative transfer within each atmospheric layer. As a result, the effect of LOS inhomogeneity on the current (1D) OMPS LP retrieval algorithm can now be studied theoretically, using realistic 3D atmospheric profiles. This work also represents a step towards enabling OMPS LP data to be ingested as part of future data assimilation efforts.

Distinctive Roles for Periplasmic Proteases in the Maintenance of Essential Outer Membrane Protein Assembly.

PubMed

Soltes, Garner R; Martin, Nicholas R; Park, Eunhae; Sutterlin, Holly A; Silhavy, Thomas J

2017-10-15

Outer membrane protein (OMP) biogenesis in Escherichia coli is a robust process essential to the life of the organism. It is catalyzed by the β-barrel assembly machine (Bam) complex, and a number of quality control factors, including periplasmic chaperones and proteases, maintain the integrity of this trafficking pathway. Little is known, however, about how periplasmic proteases recognize and degrade OMP substrates when assembly is compromised or whether different proteases recognize the same substrate at distinct points in the assembly pathway. In this work, we use well-defined assembly-defective mutants of LptD, the essential lipopolysaccharide assembly translocon, to show that the periplasmic protease DegP degrades substrates with assembly defects that prevent or impair initial contact with Bam, causing the mutant protein to accumulate in the periplasm. In contrast, another periplasmic protease, BepA, degrades a LptD mutant substrate that has engaged the Bam complex and formed a nearly complete barrel. Furthermore, we describe the role of the outer membrane lipoprotein YcaL, a protease of heretofore unknown function, in the degradation of a LptD substrate that has engaged the Bam complex but is stalled at an earlier step in the assembly process that is not accessible to BepA. Our results demonstrate that multiple periplasmic proteases monitor OMPs at distinct points in the assembly process. IMPORTANCE OMP assembly is catalyzed by the essential Bam complex and occurs in a cellular environment devoid of energy sources. Assembly intermediates that misfold can compromise this essential molecular machine. Here we demonstrate distinctive roles for three different periplasmic proteases that can clear OMP substrates with folding defects that compromise assembly at three different stages. These quality control factors help ensure the integrity of the permeability barrier that contributes to the intrinsic resistance of Gram-negative organisms to many antibiotics. Copyright © 2017 American Society for Microbiology.

The SAGE II/OSIRIS/OMPS-LP USask 2D Deseasonalized Anomaly Ozone Data Record for Use in Trend Analysis

NASA Astrophysics Data System (ADS)

Degenstein, D. A.; Bourassa, A. E.; Zawada, D.; Roth, C.; McLinden, C. A.

2017-12-01

The SAGE II/OSIRIS/OMPS-LP USask 2D ozone deseasonalized anomaly data record spans over three decades, from 1984 to the present, and has been used extensively for the determination of stratospheric ozone trends in the post Montreal Protocol era. Radiance measurements made by the three instruments have all been used to produce ozone data profiles whose native units are number density as a function of altitude. Therefore, during the merging process required to produce the extended data record it is not necessary to use meteorological data for unit conversion and all trends contained within the data record come directly from the data products themselves. Although the SAGE II occultation data record ended in 2005, both the OMPS-LP and OSIRIS limb scattered sunlight data records continue. OSIRIS has been in operation since 2001 and is well beyond its lifetime but OMPS-LP is scheduled for launch on future spacecraft so the data record should continue for many years, or even decades. It is also anticipated that SAGE III ISS data will be added to the existing record to further enhance its utility for trend analysis. This paper will outline the process used to produce the deseasonalized ozone anomaly data record detailing the issues associated with merging data records that have different biases and sampling characteristics. Issues associated with SAGE II and OSIRIS measurements that are made at different local times will also be discussed. Finally, this paper will present trend results produced using variations of the official LOTUS analysis code. These results cover an altitude range from the tropopause to 55 km and from 60 South to 60 North in ten-degree bins. It will be shown that the new OMPS-LP USask 2D data record is of excellent quality and can be used to extend the ozone data records for the purpose of trend analysis.

Sources of diversity of carbapenem resistance levels in Klebsiella pneumoniae carrying blaVIM-1.

PubMed

Loli, A; Tzouvelekis, L S; Tzelepi, E; Carattoli, A; Vatopoulos, A C; Tassios, P T; Miriagou, V

2006-09-01

To elucidate the mechanisms responsible for the diversity of beta-lactam resistance phenotypes among isolates of a VIM-1-producing Klebsiella pneumoniae (VPKP) strain that is endemic in Greek hospitals. Five VPKP clinical isolates were studied. MICs of beta-lactams were determined by agar dilution. PFGE of XbaI-digested genomic DNA was used for typing. Profiles of outer membrane proteins (OMPs) were determined by SDS-PAGE. Selected isolates were transformed with a plasmid encoding the Omp36K porin. beta-Lactamase activities were analysed by IEF and imipenem hydrolysis was assessed by spectrophotometry. VIM-1-encoding, self-transmissible plasmids were characterized by replicon typing, RFLP and hybridization with bla(VIM)- and IS26-specific probes. Characterization of integrons was performed by PCR, cloning and sequencing. Isolates exhibited highly similar PFGE patterns. Imipenem MICs were 2, 4, 16, 32 and 64 mg/L. The isolate with the highest imipenem MIC (Vipm-64) lacked a 36 kDa OMP. Expression of a cloned OmpK36 in this isolate reduced the imipenem MIC to susceptibility levels. Imipenem-hydrolysing activity was significantly higher in Vipm-16 as compared with the other isolates that expressed similar amounts of VIM-1. All isolates transferred beta-lactam resistance to Escherichia coli through conjugative, IncN plasmids that exhibited differences in the RFLP and hybridization patterns with bla(VIM)- and IS26-specific probes. The Vipm-16 plasmid, mediating the higher imipenem MICs among transconjugants, carried two copies of bla(VIM-1). Cloning and sequencing showed In-e541-like integrons truncated at the 5'CS by insertion of IS26 elements at two different positions. A VIM-1-producing strain of K. pneumoniae has evolved through OMP alterations and rearrangements in the bla(VIM-1)-carrying plasmid probably mediated by IS26, generating isolates with imipenem MICs ranging from susceptibility to resistance.

Occurrence and ecological risk assessment of organic micropollutants in the lower reaches of the Yangtze River, China: A case study of water diversion.

PubMed

Yan, Zhenhua; Yang, Haohan; Dong, Huike; Ma, Binni; Sun, Hongwei; Pan, Ting; Jiang, Runren; Zhou, Ranran; Shen, Jie; Liu, Jianchao; Lu, Guanghua

2018-08-01

Water diversion has been increasingly applied to improve water quality in many water bodies. However, little is known regarding pollution by organic micropollutants (OMPs) in water diversion projects, especially at the supplier, and this pollution may threaten the quality of transferred water. In the present study, a total of 110 OMPs belonging to seven classes were investigated in water and sediment collected from a supplier of the Yangtze River within four water diversion projects. A total of 69 and 58 target OMPs were detected in water and sediment, respectively, at total concentrations reaching 1041.78 ng/L and 5942.24 ng/g dry weight (dw). Polycyclic aromatic hydrocarbons (PAHs) and pharmaceuticals were the predominant pollutants identified. When preliminarily compared with the pollution in the receiving water, the Yangtze River generally exhibited mild OMPs pollution and good water quality parameters, implying a clean water source in the water diversion project. However, in Zongyang and Fenghuangjing, PAHs pollution was more abundant than that in the corresponding receiving water in Chaohu Lake. Ammonia nitrogen pollution in the Wangyu River was comparable to that in Taihu Lake. These findings imply that water diversion may threaten receiving waters in some cases. In addition, the risks of all detected pollutants in both water and sediment were assessed. PAHs in water, especially phenanthrene and high-molecular-weight PAHs, posed high risks to invertebrates, followed by the risks to fish and algae. Pharmaceuticals, such as antibiotics and antidepressants, may also pose risks to algae and fish at a number of locations. To the best of our knowledge, this report is the first to describe OMPs pollution in water diversion projects, and the results provide a new perspective regarding the security of water diversion projects. Copyright © 2018 Elsevier Ltd. All rights reserved.

Application of virtual phase-shifting speckle-interferometry for detection of polymorphism in the Chlamydia trachomatis omp1 gene

NASA Astrophysics Data System (ADS)

Feodorova, Valentina A.; Saltykov, Yury V.; Zaytsev, Sergey S.; Ulyanov, Sergey S.; Ulianova, Onega V.

2018-04-01

Method of phase-shifting speckle-interferometry has been used as a new tool with high potency for modern bioinformatics. Virtual phase-shifting speckle-interferometry has been applied for detection of polymorphism in the of Chlamydia trachomatis omp1 gene. It has been shown, that suggested method is very sensitive to natural genetic mutations as single nucleotide polymorphism (SNP). Effectiveness of proposed method has been compared with effectiveness of the newest bioinformatic tools, based on nucleotide sequence alignment.

Eastward shift and maintenance of Arabian Sea oxygen minimum zone: Understanding the paradox

NASA Astrophysics Data System (ADS)

Acharya, Shiba Shankar; Panigrahi, Mruganka K.

2016-09-01

The dominance of Oxygen Minimum Zone in the eastern part of the Arabian Sea (ASOMZ) instead of the more bio-productive and likely more oxygen consuming western part is the first part of the paradox. The sources of oxygen to the ASOMZ were evaluated through the distributions of different water masses using the extended optimum multiparameter (eOMP) analysis, whereas the sinks of oxygen were evaluated through the organic matter remineralization, using the apparent oxygen utilization (AOU). The contributions of major source waters to the Arabian Sea viz. Indian Deep water (dIDW), Indian Central water (ICW), Persian Gulf Water (PGW) and Red Sea Water (RSW) have been quantified through the eOMP analysis which shows that the PGW and RSW are significant for the eastward shift of ASOMZ instead of voluminous ICW and dIDW. The distribution of Net Primary Production (NPP) and AOU clearly suggest the transport of organic detritus from the highly productive western Arabian Sea to its eastern counterpart which adds to the eastward shifting of ASOMZ. A revised estimate of the seasonal variation of areal extent and volume occupied by ASOMZ through analysis of latest available data reveals a distinct intensification of ASOMZ by 30% and increase in its volume by 5% during the spring-summer transition. However, during this seasonal transition the productivity in the Arabian Sea shows 100% increase in mean NPP. This disparity between ASOMZ and monsoonal variation of productivity is the other part of the paradox, which has been constrained through apparent oxygen utilization, Net Primary Production along with a variation of core depths of source waters. This study reveals a subtle balance between the circulation of marginal oxygen-rich water masses from the western Arabian Sea and organic matter remineralization in the eastern Arabian Sea in different seasons that explains the maintenance of ASOMZ throughout the year.

The Continental Margins Program in Georgia

USGS Publications Warehouse

Cocker, M.D.; Shapiro, E.A.

1999-01-01

From 1984 to 1993, the Georgia Geologic Survey (GGS) participated in the Minerals Management Service-funded Continental Margins Program. Geological and geophysical data acquisition focused on offshore stratigraphic framework studies, phosphate-bearing Miocene-age strata, distribution of heavy minerals, near-surface alternative sources of groundwater, and development of a PC-based Coastal Geographic Information System (GIS). Seven GGS publications document results of those investigations. In addition to those publications, direct benefits of the GGS's participation include an impetus to the GGS's investigations of economic minerals on the Georgia coast, establishment of a GIS that includes computer hardware and software, and seeds for additional investigations through the information and training acquired as a result of the Continental Margins Program. These addtional investigations are quite varied in scope, and many were made possible because of GIS expertise gained as a result of the Continental Margins Program. Future investigations will also reap the benefits of the Continental Margins Program.From 1984 to 1993, the Georgia Geologic Survey (GGS) participated in the Minerals Management Service-funded Continental Margins Program. Geological and geophysical data acquisition focused on offshore stratigraphic framework studies, phosphate-bearing Miocene-age strata, distribution of heavy minerals, near-surface alternative sources of groundwater, and development of a PC-based Coastal Geographic Information System (GIS). Seven GGS publications document results of those investigations. In addition to those publications, direct benefits of the GGS's participation include an impetus to the GGS's investigations of economic minerals on the Georgia coast, establishment of a GIS that includes computer hardware and software, and seeds for additional investigations through the information and training acquired as a result of the Continental Margins Program. These additional investigations are quite varied in scope, and many were made possible because of GIS expertise gained as a result of the Continental Margins Program. Future investigations will also reap the benefits of the Continental Margins Program.

Evaluation of the Sensor Data Record from the Nadir Instruments of the Ozone Mapping Profiler Suite (OMPS)

NASA Technical Reports Server (NTRS)

Wu, Xiangqian; Liu, Quanhua; Zeng, Jian; Grotenhuis, Michael; Qian, Haifeng; Caponi, Maria; Flynn, Larry; Jaross, Glen; Sen, Bhaswar; Buss, Richard H., Jr.;

The Ozone Mapping and Profiler Suite (OMPS) Limb Profiler (LP) Version 1 aerosol extinction retrieval algorithm: theoretical basis

NASA Astrophysics Data System (ADS)

Loughman, Robert; Bhartia, Pawan K.; Chen, Zhong; Xu, Philippe; Nyaku, Ernest; Taha, Ghassan

2018-05-01

The theoretical basis of the Ozone Mapping and Profiler Suite (OMPS) Limb Profiler (LP) Version 1 aerosol extinction retrieval algorithm is presented. The algorithm uses an assumed bimodal lognormal aerosol size distribution to retrieve aerosol extinction profiles at 675 nm from OMPS LP radiance measurements. A first-guess aerosol extinction profile is updated by iteration using the Chahine nonlinear relaxation method, based on comparisons between the measured radiance profile at 675 nm and the radiance profile calculated by the Gauss-Seidel limb-scattering (GSLS) radiative transfer model for a spherical-shell atmosphere. This algorithm is discussed in the context of previous limb-scattering aerosol extinction retrieval algorithms, and the most significant error sources are enumerated. The retrieval algorithm is limited primarily by uncertainty about the aerosol phase function. Horizontal variations in aerosol extinction, which violate the spherical-shell atmosphere assumed in the version 1 algorithm, may also limit the quality of the retrieved aerosol extinction profiles significantly.

Molecular transformers in the cell: lessons learned from the DegP protease-chaperone.

PubMed

Sawa, Justyna; Heuck, Alexander; Ehrmann, Michael; Clausen, Tim

2010-04-01

Structure-function analysis of DegP revealed a novel mechanism for protease and chaperone regulation. Binding of unfolded proteins induces the oligomer reassembly from the resting hexamer (DegP6) into the functional protease-chaperone DegP12/24. The newly formed cage exhibits the characteristics of a proteolytic folding chamber, shredding those proteins that are severely misfolded while stabilizing and protecting proteins present in their native state. Isolation of native DegP complexes with folded outer membrane proteins (OMPs) highlights the importance of DegP in OMP biogenesis. The encapsulated OMP beta-barrel is significantly stabilized in the hydrophobic chamber of DegP12/24 and thus DegP seems to employ a reciprocal mechanism to those chaperones assisting the folding of water soluble proteins via polar interactions. In addition, we discuss in this review similarities to other complex proteolytic machines that, like DegP, are under control of a substrate-induced or stress-induced oligomer conversion.

General Method to Determine the Flux of Charged Molecules through Nanopores Applied to β-Lactamase Inhibitors and OmpF.

PubMed

Ghai, Ishan; Pira, Alessandro; Scorciapino, Mariano Andrea; Bodrenko, Igor; Benier, Lorraine; Ceccarelli, Matteo; Winterhalter, Mathias; Wagner, Richard

2017-03-16

A major challenge in the discovery of the new antibiotics against Gram-negative bacteria is to achieve sufficiently fast permeation in order to avoid high doses causing toxic side effects. So far, suitable assays for quantifying the uptake of charged antibiotics into bacteria are lacking. We apply an electrophysiological zero-current assay using concentration gradients of β-lactamase inhibitors combined with single-channel conductance to quantify their flux rates through OmpF. Molecular dynamic simulations provide in addition details on the interactions between the nanopore wall and the charged solutes. In particular, the interaction barrier for three β-lactamase inhibitors is surprisingly as low as 3-5 kcal/mol and only slightly above the diffusion barrier of ions such as chloride. Within our macroscopic constant field model, we determine that at a zero-membrane potential a concentration gradient of 10 μM of avibactam, sulbactam, or tazobactam can create flux rates of roughly 620 molecules/s per OmpF trimer.

Typing and comparative genome analysis of Brucella melitensis isolated from Lebanon.

PubMed

Abou Zaki, Natalia; Salloum, Tamara; Osman, Marwan; Rafei, Rayane; Hamze, Monzer; Tokajian, Sima

2017-10-16

Brucella melitensis is the main causative agent of the zoonotic disease brucellosis. This study aimed at typing and characterizing genetic variation in 33 Brucella isolates recovered from patients in Lebanon. Bruce-ladder multiplex PCR and PCR-RFLP of omp31, omp2a and omp2b were performed. Sixteen representative isolates were chosen for draft-genome sequencing and analyzed to determine variations in virulence, resistance, genomic islands, prophages and insertion sequences. Comparative whole-genome single nucleotide polymorphism analysis was also performed. The isolates were confirmed to be B. melitensis. Genome analysis revealed multiple virulence determinants and efflux pumps. Genome comparisons and single nucleotide polymorphisms divided the isolates based on geographical distribution but revealed high levels of similarity between the strains. Sequence divergence in B. melitensis was mainly due to lateral gene transfer of mobile elements. This is the first report of an in-depth genomic characterization of B. melitensis in Lebanon. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Outer membrane changes in Pseudomonas stutzeri resistant to chlorhexidine diacetate and cetylpyridinium chloride.

PubMed

Tattawasart, U; Maillard, J Y; Furr, J R; Russell, A D

2000-11-01

Changes in outer membrane proteins (OMP) and lipopolysaccharide (LPS) in cells of strains of Pseudomonas stutzeri sensitive and resistant to chlorhexidine diacetate (CHA) or cetylpyridinium chloride (CPC) have been examined. Four of five CHA-resistant strains had alterations in OMP profiles, including the expression of two additional protein bands. All the CPC-resistant strains had altered OMP profiles but the changes varied from strain to strain. Loss of the fastest-migrating bands was observed in the LPS from CHX-resistant strains. In strain JM 302R with high-level CHX resistance (minimal inhibitory concentration 100 mg/l as opposed to 2.5 mg/l in the parent-strain, JM 302), all the fast-migrating bands were lost and the strain showed 'cross-resistance' to polymyxin, gentamicin and ethylenediamine tetraacetic acid. It is however, possible that the altered LPS patterns reflect a response to alterations in other components rather than being directly associated per se with the enhanced resistance.

Phylogenetic Variants of Rickettsia africae, and Incidental Identification of "Candidatus Rickettsia Moyalensis" in Kenya.

PubMed

Kimita, Gathii; Mutai, Beth; Nyanjom, Steven Ger; Wamunyokoli, Fred; Waitumbi, John

2016-07-01

Rickettsia africae, the etiological agent of African tick bite fever, is widely distributed in sub-Saharan Africa. Contrary to reports of its homogeneity, a localized study in Asembo, Kenya recently reported high genetic diversity. The present study aims to elucidate the extent of this heterogeneity by examining archived Rickettsia africae DNA samples collected from different eco-regions of Kenya. To evaluate their phylogenetic relationships, archived genomic DNA obtained from 57 ticks a priori identified to contain R. africae by comparison to ompA, ompB and gltA genes was used to amplify five rickettsial genes i.e. gltA, ompA, ompB, 17kDa and sca4. The resulting amplicons were sequenced. Translated amino acid alignments were used to guide the nucleotide alignments. Single gene and concatenated alignments were used to infer phylogenetic relationships. Out of the 57 DNA samples, three were determined to be R. aeschlimanii and not R. africae. One sample turned out to be a novel rickettsiae and an interim name of "Candidatus Rickettsia moyalensis" is proposed. The bonafide R. africae formed two distinct clades. Clade I contained 9% of the samples and branched with the validated R. africae str ESF-5, while clade II (two samples) formed a distinct sub-lineage. This data supports the use of multiple genes for phylogenetic inferences. It is determined that, despite its recent emergence, the R. africae lineage is diverse. This data also provides evidence of a novel Rickettsia species, Candidatus Rickettsia moyalensis.

Release 2 data products from the Ozone Mapping and Profiler Suite (OMPS) Limb Profiler

NASA Astrophysics Data System (ADS)

Xu, Philippe Q.; Bhartia, Pawan K.; Jaross, Glen R.; DeLand, Matthew T.; Larsen, Jack C.; Fleig, Albert; Kahn, Daniel; Zhu, Tong; Chen, Zhong; Gorkavyi, Nick; Warner, Jeremy; Linda, Michael; Chen, Hong G.; Kowitt, Mark; Haken, Michael; Hall, Peter

2014-10-01

The OMPS Limb Profiler (LP) was launched on board the NASA Suomi National Polar-orbiting Partnership (SNPP) satellite in October 2011. OMPS-LP is a limb-scattering hyperspectral sensor that provides ozone profiling capability at 1.8 km vertical resolution from cloud top to 60 km altitude. The use of three parallel slits allows global coverage in approximately four days. We have recently completed a full reprocessing of all LP data products, designated as Release 2, that improves the accuracy and quality of these products. Level 1 gridded radiance (L1G) changes include intra-orbit and seasonal correction of variations in wavelength registration, revised static and intra-orbit tangent height adjustments, and simplified pixel selection from multiple images. Ozone profile retrieval changes include removal of the explicit aerosol correction, exclusion of channels contaminated by stratospheric OH emission, a revised instrument noise characterization, improved synthetic solar spectrum, improved pressure and temperature ancillary data, and a revised ozone climatology. Release 2 data products also include aerosol extinction coefficient profiles derived with the prelaunch retrieval algorithm. Our evaluation of OMPS LP Release 2 data quality is good. Zonal average ozone profile comparisons with Aura MLS data typically show good agreement, within 5-10% over the altitude range 20-50 km between 60°S and 60°N. The aerosol profiles agree well with concurrent satellite measurements such as CALIPSO and OSIRIS, and clearly detect exceptional events such as volcanic eruptions and the Chelyabinsk bolide in February 2013.

Release 2 data products from the Ozone Mapping and Profiler Suite (OMPS) Limb Profiler

NASA Technical Reports Server (NTRS)

Xu, Q. Philippe; Bhartia, Pawan K.; Jaross, Glen R.; Deland, Matthew T.; Larsen, Jack C.; Fleig, Albert; Kahn, Daniel; Zhu, Tong; Chen, Zhong; Gorkavyi, Nick;

How β-Lactam Antibiotics Enter Bacteria: A Dialogue with the Porins

PubMed Central

Molitor, Alexander; Bolla, Jean-Michel; Bessonov, Andrey N.; Winterhalter, Mathias; Pagès, Jean-Marie

2009-01-01

Background Multi-drug resistant (MDR) infections have become a major concern in hospitals worldwide. This study investigates membrane translocation, which is the first step required for drug action on internal bacterial targets. β-lactams, a major antibiotic class, use porins to pass through the outer membrane barrier of Gram-negative bacteria. Clinical reports have linked the MDR phenotype to altered membrane permeability including porin modification and efflux pump expression. Methodology/Principal Findings Here influx of β-lactams through the major Enterobacter aerogenes porin Omp36 is characterized. Conductance measurements through a single Omp36 trimer reconstituted into a planar lipid bilayer allowed us to count the passage of single β-lactam molecules. Statistical analysis of each transport event yielded the kinetic parameters of antibiotic travel through Omp36 and distinguishable translocation properties of β-lactams were quantified for ertapenem and cefepime. Expression of Omp36 in an otherwise porin-null bacterial strain is shown to confer increases in the killing rate of these antibiotics and in the corresponding bacterial susceptibility. Conclusions/Significance We propose the idea of a molecular “passport” that allows rapid transport of substrates through porins. Deciphering antibiotic translocation provides new insights for the design of novel drugs that may be highly effective at passing through the porin constriction zone. Such data may hold the key for the next generation of antibiotics capable of rapid intracellular accumulation to circumvent the further development MDR infections. PMID:19434239

An adaptive response of Enterobacter aerogenes to imipenem: regulation of porin balance in clinical isolates.

PubMed

Lavigne, Jean-Philippe; Sotto, Albert; Nicolas-Chanoine, Marie-Hélène; Bouziges, Nicole; Pagès, Jean-Marie; Davin-Regli, Anne

2013-02-01

Imipenem (IPM) is a carbapenem antibiotic frequently used in severe hospital infections. Several reports have mentioned the emergence of resistant isolates exhibiting membrane modifications. A study was conducted between September 2005 and August 2007 to survey infections due to Enterobacter aerogenes in patients hospitalised in a French university hospital. Resistant E. aerogenes clinical isolates obtained from patients treated with IPM and collected during the 3 months following initiation of treatment were phenotypically and molecularly characterised for β-lactamases, efflux pumps activity and outer membrane proteins. Among the 339 patients infected with E. aerogenes during the study period, 41 isolates (12.1%) were resistant to extended-spectrum cephalosporins and 17 patients (5.0%) were treated with IPM. The isolates from these 17 patients presented TEM-24 and basal efflux expression. Following IPM treatment, an IPM-intermediate-susceptible (IPM-I) isolate emerged in 11 patients and an IPM-resistant (IPM-R) isolate in 6 patients. A change in the porin balance (Omp35/Omp36) was observed in IPM-I isolates exhibiting ertapenem resistance. Finally, a porin deficiency (Omp35 and Omp36 absence) was detected in IPM-R isolates associated with efflux pump expression. This study indicates that the alteration in porin expression, including the shift of porin expression and lack of porins, contribute to the E. aerogenes adaptive response to IPM treatment. Copyright © 2012 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.

Experimental validation of the predicted binding site of Escherichia coli K1 outer membrane protein A to human brain microvascular endothelial cells: identification of critical mutations that prevent E. coli meningitis.

PubMed

Pascal, Tod A; Abrol, Ravinder; Mittal, Rahul; Wang, Ying; Prasadarao, Nemani V; Goddard, William A

2010-11-26

Escherichia coli K1, the most common cause of meningitis in neonates, has been shown to interact with GlcNAc1-4GlcNAc epitopes of Ecgp96 on human brain microvascular endothelial cells (HBMECs) via OmpA (outer membrane protein A). However, the precise domains of extracellular loops of OmpA interacting with the chitobiose epitopes have not been elucidated. We report the loop-barrel model of these OmpA interactions with the carbohydrate moieties of Ecgp96 predicted from molecular modeling. To test this model experimentally, we generated E. coli K1 strains expressing OmpA with mutations of residues predicted to be critical for interaction with the HBMEC and tested E. coli invasion efficiency. For these same mutations, we predicted the interaction free energies (including explicit calculation of the entropy) from molecular dynamics (MD), finding excellent correlation (R(2) = 90%) with experimental invasion efficiency. Particularly important is that mutating specific residues in loops 1, 2, and 4 to alanines resulted in significant inhibition of E. coli K1 invasion in HBMECs, which is consistent with the complete lack of binding found in the MD simulations for these two cases. These studies suggest that inhibition of the interactions of these residues of Loop 1, 2, and 4 with Ecgp96 could provide a therapeutic strategy to prevent neonatal meningitis due to E. coli K1.

OMPS Limb Profiler Instrument Performance Assessment

NASA Technical Reports Server (NTRS)

Jaross, Glen R.; Bhartia, Pawan K.; Chen, Grace; Kowitt, Mark; Haken, Michael; Chen, Zhong; Xu, Philippe; Warner, Jeremy; Kelly, Thomas

2014-01-01

Following the successful launch of the Ozone Mapping and Profiler Suite (OMPS) aboard the Suomi National Polar-orbiting Partnership (SNPP) spacecraft, the NASA OMPS Limb team began an evaluation of instrument and data product performance. The focus of this paper is the instrument performance in relation to the original design criteria. Performance that is closer to expectations increases the likelihood that limb scatter measurements by SNPP OMPS and successor instruments can form the basis for accurate long-term monitoring of ozone vertical profiles. The team finds that the Limb instrument operates mostly as designed and basic performance meets or exceeds the original design criteria. Internally scattered stray light and sensor pointing knowledge are two design challenges with the potential to seriously degrade performance. A thorough prelaunch characterization of stray light supports software corrections that are accurate to within 1% in radiances up to 60 km for the wavelengths used in deriving ozone. Residual stray light errors at 1000nm, which is useful in retrievals of stratospheric aerosols, currently exceed 10%. Height registration errors in the range of 1 km to 2 km have been observed that cannot be fully explained by known error sources. An unexpected thermal sensitivity of the sensor also causes wavelengths and pointing to shift each orbit in the northern hemisphere. Spectral shifts of as much as 0.5nm in the ultraviolet and 5 nm in the visible, and up to 0.3 km shifts in registered height, must be corrected in ground processing.

On the Five-Moment Hamburger Maximum Entropy Reconstruction

NASA Astrophysics Data System (ADS)

Summy, D. P.; Pullin, D. I.

2018-05-01

We consider the Maximum Entropy Reconstruction (MER) as a solution to the five-moment truncated Hamburger moment problem in one dimension. In the case of five monomial moment constraints, the probability density function (PDF) of the MER takes the form of the exponential of a quartic polynomial. This implies a possible bimodal structure in regions of moment space. An analytical model is developed for the MER PDF applicable near a known singular line in a centered, two-component, third- and fourth-order moment (μ _3 , μ _4 ) space, consistent with the general problem of five moments. The model consists of the superposition of a perturbed, centered Gaussian PDF and a small-amplitude packet of PDF-density, called the outlying moment packet (OMP), sitting far from the mean. Asymptotic solutions are obtained which predict the shape of the perturbed Gaussian and both the amplitude and position on the real line of the OMP. The asymptotic solutions show that the presence of the OMP gives rise to an MER solution that is singular along a line in (μ _3 , μ _4 ) space emanating from, but not including, the point representing a standard normal distribution, or thermodynamic equilibrium. We use this analysis of the OMP to develop a numerical regularization of the MER, creating a procedure we call the Hybrid MER (HMER). Compared with the MER, the HMER is a significant improvement in terms of robustness and efficiency while preserving accuracy in its prediction of other important distribution features, such as higher order moments.

Evaluation of recombinant porin (rOmp2a) protein as a potential antigen candidate for serodiagnosis of Human Brucellosis.

PubMed

Pathak, Prachi; Kumar, Ashu; Thavaselvam, Duraipandian

2017-07-11

Brucellosis is an important zoonotic disease caused by different Brucella species and human brucellosis is commonly prevalent in different states of India. Among various Brucella species, B. melitensis is most pathogenic to human and included as category B biothreat which can cause infection through aerosol, cut, wounds in skin and contact with infected animals. The diagnosis of human brucellosis is very important for proper treatment and management of disease as there is no vaccine available for human use. The present study was designed to clone, express and purify immunodominant recombinant omp2a (rOmp2a) porin protein of B. melitensis and to evaluate this new antigen candidate for specific serodiagnosis of human brucellosis by highly sensitive iELISA (indirect enzyme linked immunosorbent assay). Omp2a gene of B. melitensis 16 M strain was cloned and expressed in pET-SUMO expression system. The recombinant protein was purified under denaturing conditions using 8 M urea. The purified recombinant protein was confirmed by western blotting by reacting with anti-HIS antibody. The sero-reactivity of the recombinant protein was also checked by reacting with antisera of experimentally infected mice with B. melitensis 16 M at different time points. Serodiagnostic potential of recombinant porin antigen was tested against 185 clinical serum samples collected from regions endemic to brucellosis in southern part of India by iELISA. The samples were grouped into five groups. Group 1 contained cultured confirmed positive serum samples of brucellosis (n = 15), group 2 contained sera samples from positive cases of brucellosis previously tested by conventional methods of RBPT (n = 28) and STAT (n = 26), group 3 contained sera samples negative by RBPT(n = 36) and STAT (n = 32), group 4 contained sera samples of other febrile illness and PUO case (n = 35) and group 5 contained confirmed negative sera samples from healthy donors (n = 23). The rOmp2a was found to be immunoreactive by iELISA and western blotting. The test showed a sensitivity of 93.75% and specificity of 95.83% when tested against 185 serum samples. For determination of statistical significance between experimental groups and control groups, Student's t test was performed on the data. Omp2a emerges as a potential antigen candidate for serodiagnosis of human brucellosis.

Comparative diagnostic evaluation of OMP31 gene based TaqMan® real-time PCR assay with visual LAMP assay and indirect ELISA for caprine brucellosis.

PubMed

Saini, Suman; Gupta, V K; Gururaj, K; Singh, D D; Pawaiya, R V S; Gangwar, N K; Mishra, A K; Dwivedi, Deepak; Andani, Dimple; Kumar, Ashok; Goswami, T K

2017-08-01

Brucellosis is one of the leading causes of abortion in domestic animals that imposes costs on both economy and society. The disease is highly zoonotic and poses risk to animal handlers due to its zoonotic nature. It causes stillbirth, loss of kids and abortion in last term of pregnancy. Reproductive damage includes infertility in does and orchitis and epididymitis in breeding bucks, which result in high financial losses to farmers and the agriculture industry as a whole. It requires highly sensitive and specific assays to diagnose the disease at field level. In the current study, a visual loop-mediated isothermal amplification (LAMP) assay and the TaqMan® real-time PCR were developed with high sensitivity and specificity. For the TaqMan® probe, real-time PCR primers were developed using Omp31 gene as target and primers were designed using discontiguous conserved sequences of Omp31 gene. The Omp31 probes were designed by attaching 6-FAM reporter dye at the 5' end and BHQ-1 quencher at the 3' end. Published primers were used for visual LAMP assay targeting the Omp25 gene. Sensitivity of the standardized visual LAMP assay and TaqMan® real-time PCR assay was determined by serial dilution of positive Brucella melitensis DNA (10 2 to 10 -4  ng) obtained from standard culture. The TaqMan® probe real-time assay can detect as low as 100 fg of B. melitensis DNA, whereas culture from vaginal swab washings has a limit of detection (LOD) of only 1 cfu/ml. Similarly, the visual LAMP assay can detect as low as 10 fg of B. melitensis DNA as compared to an LOD of 30 cfu/ml from culture of vaginal swab washings. Both assays were compared with serological tests (serum tube agglutination test (STAT) and indirect enzyme-linked immunosorbent assay (iELISA)) for diagnostic sensitivity and specificity. Diagnostic sensitivities and specificities for TaqMan® real-time PCR vs. LAMP assays were 98 and 100% vs. 100 and 97.8%, respectively. Results of visual LAMP assay indicated that LAMP is a fast, specific, sensitive, inexpensive and suitable method for diagnosis of B. melitensis infection under field conditions. On the other hand, Omp31 TaqMan® probe real-time assay can be used in conjunction with the other field-based diagnostic tests due to its high specificity.

Detecting and quantifying stellar magnetic fields. Sparse Stokes profile approximation using orthogonal matching pursuit

NASA Astrophysics Data System (ADS)

Carroll, T. A.; Strassmeier, K. G.

2014-03-01

Context. In recent years, we have seen a rapidly growing number of stellar magnetic field detections for various types of stars. Many of these magnetic fields are estimated from spectropolarimetric observations (Stokes V) by using the so-called center-of-gravity (COG) method. Unfortunately, the accuracy of this method rapidly deteriorates with increasing noise and thus calls for a more robust procedure that combines signal detection and field estimation. Aims: We introduce an estimation method that provides not only the effective or mean longitudinal magnetic field from an observed Stokes V profile but also uses the net absolute polarization of the profile to obtain an estimate of the apparent (i.e., velocity resolved) absolute longitudinal magnetic field. Methods: By combining the COG method with an orthogonal-matching-pursuit (OMP) approach, we were able to decompose observed Stokes profiles with an overcomplete dictionary of wavelet-basis functions to reliably reconstruct the observed Stokes profiles in the presence of noise. The elementary wave functions of the sparse reconstruction process were utilized to estimate the effective longitudinal magnetic field and the apparent absolute longitudinal magnetic field. A multiresolution analysis complements the OMP algorithm to provide a robust detection and estimation method. Results: An extensive Monte-Carlo simulation confirms the reliability and accuracy of the magnetic OMP approach where a mean error of under 2% is found. Its full potential is obtained for heavily noise-corrupted Stokes profiles with signal-to-noise variance ratios down to unity. In this case a conventional COG method yields a mean error for the effective longitudinal magnetic field of up to 50%, whereas the OMP method gives a maximum error of 18%. It is, moreover, shown that even in the case of very small residual noise on a level between 10-3 and 10-5, a regime reached by current multiline reconstruction techniques, the conventional COG method incorrectly interprets a large portion of the residual noise as a magnetic field, with values of up to 100 G. The magnetic OMP method, on the other hand, remains largely unaffected by the noise, regardless of the noise level the maximum error is no greater than 0.7 G.

Immunization of cows with novel core glycolipid vaccine induces anti-endotoxin antibodies in bovine colostrum.

PubMed

Cross, Alan S; Karreman, Hubert J; Zhang, Lei; Rosenberg, Zeil; Opal, Steven M; Lees, Andrew

2014-10-21

Translocation of gut-derived Gram-negative bacterial (GNB) lipopolysaccharide (LPS, or endotoxin) is a source of systemic inflammation that exacerbates HIV, cardiovascular and gastrointestinal diseases and malnutrition. The oral administration of bovine colostrum (BC) reduces endotoxemia in patients with impaired gut barrier function. Consequently, BC enriched in antibodies to LPS may ameliorate endotoxemia-related morbidities. We developed a detoxified J5 LPS/group B meningococcal outer membrane protein (J5dLPS/OMP) vaccine that induces antibodies against a highly conserved core region of LPS and protects against heterologous GNB infection. We now examine the ability of this vaccine to elicit anti-core endotoxin antibodies in BC. Two cohorts of pregnant cows were immunized with this vaccine in combination with FICA (Cohort 1) or Emulsigen-D (Cohort 2) adjuvants. Antibody responses to the J5 core LPS antigen were measured in both serum and colostrum and compared to antibody levels elicited by a commercially available veterinary vaccine (J5 Bacterin) comprised of heat-killed Escherichia coli O111, J5 mutant bacteria, from which the J5 LPS was purified. The J5dLPS/OMP vaccine induced high titers of serum IgG antibody to J5 LPS in all seven cows. Both IgG and to a lesser extent IgA anti-J5 LPS antibodies were generated in the colostrum. The J5dLPS/OMP vaccine was significantly more immunogenic in mice than was the J5 Bacterin. BC enriched in anti-J5 LPS antibody reduced circulating endotoxin levels in neutropenic rats, a model of "leaky gut". The J5dLPS/OMP vaccine elicits high titers of serum anti-endotoxin antibodies in cows that is passed to the colostrum. This BC enriched in anti-core LPS antibodies has the potential to reduce endotoxemia and ameliorate endotoxin-related systemic inflammation in patients with impaired gut barrier function. Since this vaccine is significantly more immunogenic than the J5 Bacterin vaccine, this J5dLPS/OMP vaccine might prove to be more useful for veterinary indications as well. Copyright © 2014 Elsevier Ltd. All rights reserved.

Extending the NOAA SBUV(/2) Ozone Profile Record

NASA Astrophysics Data System (ADS)

Frith, S. M.; Wild, J.; Long, C. S.

2017-12-01

Since the signing of the Montreal Protocol in 1987 and its subsequent agreements banning anthropogenic ozone depleting substances (ODS) the climate community has been anticipating the ability to detect the recovery of the ozone layer. This recovery is complicated by climate changes associated with the increase of CO2 in the both the troposphere and stratosphere. The Climate Prediction Center (CPC) has generated a long term total column and profile ozone climate data record (CDR) based on the SBUV and SBUV/2 on Nimbus 7 and the NOAA Polar Orbiting Environmental Satellites (POES): NOAA-9, -11, -14, -16, -17, -18 and -19 spanning 38 years from 1978 to 2016. This dataset uses observations from a single instrument for each time period and an adjustment scheme to remove inter-satellite differences. The last of these SBUV/2 instruments resides on NOAA-19 launched in 2009, and with drifting equatorial crossing time will soon loose latitudinal coverage, and be impacted by an increasing solar zenith angle. The Ozone Mapping and Profiler Suite (OMPS) instrument has replaced the SBUV/2 as the primary ozone monitoring instrument at NOAA. It is taking observations on the Suomi-NPOESS Preparatory Project (S-NPP) satellite which was launched in 2011 and will be on future JPSS satellites. JPSS-1 is expected to be launched in late 2017, and later JPSS satellites will additionally carry the OMPS instrument. Reprocessed OMPS Nadir Profile (NP) and Nadir Mapper (NM) level 2 data has been made available by NESDIS/STAR covering the period from 2012 through 2016. The OMPS NP has been characterized and calibrated to be very similar to the SBUV/2. Results of extending the SBUV(/2) dataset with ozone profile data from OMPS will be reviewed. Stability of ozone recovery trend estimates using these datasets will be explored using the Hockey Stick approach of Reinsel (2002) near-globally (50N-50S), tropically and at mid-latitudes. Seasonality of the trend results will be examined. Reinsel, G.C., et al Journal of Geophys. Res., 107, p4078 (2002).

VDAC electronics: 1. VDAC-hexo(gluco)kinase generator of the mitochondrial outer membrane potential.

PubMed

Lemeshko, Victor V

2014-05-01

The simplest mechanism of the generation of the mitochondrial outer membrane potential (OMP) by the VDAC (voltage-dependent anion channel)-hexokinase complex (VHC), suggested earlier, and by the VDAC-glucokinase complex (VGC), was computationally analyzed. Even at less than 4% of VDACs bound to hexokinase, the calculated OMP is high enough to trigger the electrical closure of VDACs beyond the complexes at threshold concentrations of glucose. These results confirmed our previous hypothesis that the Warburg effect is caused by the electrical closure of VDACs, leading to global restriction of the outer membrane permeability coupled to aerobic glycolysis. The model showed that the inhibition of the conductance and/or an increase in the voltage sensitivity of a relatively small fraction of VDACs by factors like tubulin potentiate the electrical closure of the remaining free VDACs. The extrusion of calcium ions from the mitochondrial intermembrane space by the generated OMP, positive inside, might increase cancer cell resistance to death. Within the VGC model, the known effect of induction of ATP release from mitochondria by accumulated glucose-6-phosphate in pancreatic beta cells might result not only of the known effect of GK dissociation from the VDAC-GK complex, but also of a decrease in the free energy of glucokinase reaction, leading to the OMP decrease and VDAC opening. We suggest that the VDAC-mediated electrical control of the mitochondrial outer membrane permeability, dependent on metabolic conditions, is a fundamental physiological mechanism of global regulation of mitochondrial functions and of cell death. Copyright © 2014 Elsevier B.V. All rights reserved.

Haemophilus ducreyi Outer Membrane Determinants, Including DsrA, Define Two Clonal Populations

PubMed Central

White, Catherine Dinitra; Leduc, Isabelle; Olsen, Bonnie; Jeter, Chrystina; Harris, Chavala; Elkins, Christopher

2005-01-01

The Haemophilus ducreyi outer membrane component DsrA (for ducreyi serum resistance A) is necessary for complete resistance to normal human serum (NHS). When DsrA expression in 19 temporally and geographically diverse clinical isolates of H. ducreyi was examined by Western blotting, 5 of the strains expressed a different immunotype of the DsrA protein (DsrAII) than the well-characterized prototypical strain 35000HP (DsrAI). The predicted DsrA proteins expressed by the DsrAII strains were 100% identical to each other but only 48% identical to that of strain 35000HP. In addition to the DsrAII protein, class II strains also expressed variant forms of other outer membrane proteins (OMPs) including NcaA (necessary for collagen adhesion A), DltA (ducreyi lectin A), Hlp (H. ducreyi lipoprotein), major OMP, and/or OmpA2 (for OMP A2) and synthesized a distinct, faster-migrating lipooligosaccharide. Based on these data, strains expressing DsrAI were termed class I, and those expressing DsrAII were termed class II. Expression of dsrAII from strain CIP 542 ATCC in the class I dsrAI mutant FX517 (35000HP background), which does not express a DsrA protein, rendered this strain resistant to 50% NHS. This demonstrates that DsrAII protein is also critical to serum resistance. Taken together, these results indicate that there are two clonal populations of H. ducreyi. The implications of two classes of H. ducreyi strains differing in important antigenic outer membrane components are discussed. PMID:15784585

Merged SAGE II, Ozone_cci and OMPS ozone profiles dataset and evaluation of ozone trends in the stratosphere

NASA Astrophysics Data System (ADS)

Tamminen, J.; Sofieva, V.; Kyrölä, E.; Laine, M.; Degenstein, D. A.; Bourassa, A. E.; Roth, C.; Zawada, D.; Weber, M.; Rozanov, A.; Rahpoe, N.; Stiller, G. P.; Laeng, A.; von Clarmann, T.; Walker, K. A.; Sheese, P.; Hubert, D.; Van Roozendael, M.; Zehner, C.; Damadeo, R. P.; Zawodny, J. M.; Kramarova, N. A.; Bhartia, P. K.

2017-12-01

We present a merged dataset of ozone profiles from several satellite instruments: SAGE II on ERBS, GOMOS, SCIAMACHY and MIPAS on Envisat, OSIRIS on Odin, ACE-FTS on SCISAT, and OMPS on Suomi-NPP. The merged dataset is created in the framework of European Space Agency Climate Change Initiative (Ozone_cci) with the aim of analyzing stratospheric ozone trends. For the merged dataset, we used the latest versions of the original ozone datasets. The datasets from the individual instruments have been extensively validated and inter-compared; only those datasets, which are in good agreement and do not exhibit significant drifts with respect to collocated ground-based observations and with respect to each other, are used for merging. The long-term SAGE-CCI-OMPS dataset is created by computation and merging of deseasonalized anomalies from individual instruments. The merged SAGE-CCI-OMPS dataset consists of deseasonalized anomalies of ozone in 10° latitude bands from 90°S to 90°N and from 10 to 50 km in steps of 1 km covering the period from October 1984 to July 2016. This newly created dataset is used for evaluating ozone trends in the stratosphere through multiple linear regression. Negative ozone trends in the upper stratosphere are observed before 1997 and positive trends are found after 1997. The upper stratospheric trends are statistically significant at mid-latitudes in the upper stratosphere and indicate ozone recovery, as expected from the decrease of stratospheric halogens that started in the middle of the 1990s.

Is anaerobic digestion effective for the removal of organic micropollutants and biological activities from sewage sludge?

PubMed

Gonzalez-Gil, L; Papa, M; Feretti, D; Ceretti, E; Mazzoleni, G; Steimberg, N; Pedrazzani, R; Bertanza, G; Lema, J M; Carballa, M

2016-10-01

The occurrence of emerging organic micropollutants (OMPs) in sewage sludge has been widely reported; nevertheless, their fate during sludge treatment remains unclear. The objective of this work was to study the fate of OMPs during mesophilic and thermophilic anaerobic digestion (AD), the most common processes used for sludge stabilization, by using raw sewage sludge without spiking OMPs. Moreover, the results of analytical chemistry were complemented with biological assays in order to verify the possible adverse effects (estrogenic and genotoxic) on the environment and human health in view of an agricultural (re)use of digested sludge. Musk fragrances (AHTN, HHCB), ibuprofen (IBP) and triclosan (TCS) were the most abundant compounds detected in sewage sludge. In general, the efficiency of the AD process was not dependent on operational parameters but compound-specific: some OMPs were highly biotransformed (e.g. sulfamethoxazole and naproxen), while others were only slightly affected (e.g. IBP and TCS) or even unaltered (e.g. AHTN and HHCB). The MCF-7 assay evidenced that estrogenicity removal was driven by temperature. The Ames test did not show point mutation in Salmonella typhimurium while the Comet test exhibited a genotoxic effect on human leukocytes attenuated by AD. This study highlights the importance of combining chemical analysis and biological activities in order to establish appropriate operational strategies for a safer disposal of sewage sludge. Actually, it was demonstrated that temperature has an insignificant effect on the disappearance of the parent compounds while it is crucial to decrease estrogenicity. Copyright © 2016 Elsevier Ltd. All rights reserved.

Brucella abortus Inhibits Major Histocompatibility Complex Class II Expression and Antigen Processing through Interleukin-6 Secretion via Toll-Like Receptor 2▿

PubMed Central

Barrionuevo, Paula; Cassataro, Juliana; Delpino, M. Victoria; Zwerdling, Astrid; Pasquevich, Karina A.; Samartino, Clara García; Wallach, Jorge C.; Fossati, Carlos A.; Giambartolomei, Guillermo H.

2008-01-01

The strategies that allow Brucella abortus to survive inside macrophages for prolonged periods and to avoid the immunological surveillance of major histocompatibility complex class II (MHC-II)-restricted gamma interferon (IFN-γ)-producing CD4+ T lymphocytes are poorly understood. We report here that infection of THP-1 cells with B. abortus inhibited expression of MHC-II molecules and antigen (Ag) processing. Heat-killed B. abortus (HKBA) also induced both these phenomena, indicating the independence of bacterial viability and involvement of a structural component of the bacterium. Accordingly, outer membrane protein 19 (Omp19), a prototypical B. abortus lipoprotein, inhibited both MHC-II expression and Ag processing to the same extent as HKBA. Moreover, a synthetic lipohexapeptide that mimics the structure of the protein lipid moiety also inhibited MHC-II expression, indicating that any Brucella lipoprotein could down-modulate MHC-II expression and Ag processing. Inhibition of MHC-II expression and Ag processing by either HKBA or lipidated Omp19 (L-Omp19) depended on Toll-like receptor 2 and was mediated by interleukin-6. HKBA or L-Omp19 also inhibited MHC-II expression and Ag processing of human monocytes. In addition, exposure to the synthetic lipohexapeptide inhibited Ag-specific T-cell proliferation and IFN-γ production of peripheral blood mononuclear cells from Brucella-infected patients. Together, these results indicate that there is a mechanism by which B. abortus may prevent recognition by T cells to evade host immunity and establish a chronic infection. PMID:17984211

Expression of calmodulin mRNA in rat olfactory neuroepithelium.

PubMed

Biffo, S; Goren, T; Khew-Goodall, Y S; Miara, J; Margolis, F L

1991-04-01

A calmodulin (CaM) cDNA was isolated by differential hybridization screening of a lambda gt10 library prepared from rat olfactory mucosa. This cDNA fragment, containing most of the open reading frame of the rat CaMI gene, was subcloned and used to characterize steady-state expression of CaM mRNA in rat olfactory neuroepithelium and bulb. Within the bulb mitral cells are the primary neuronal population expressing CaM mRNA. The major CaM mRNA expressed in the olfactory mucosa is 1.7 kb with smaller contributions from mRNAs of 4.0 and 1.4 kb. CaM mRNA was primarily associated with the olfactory neurons and, despite the cellular complexity of the tissue and the known involvement of CaM in diverse cellular processes, was only minimally evident in sustentacular cells, gland cells or respiratory epithelium. Following bulbectomy CaM mRNA declines in the olfactory neuroepithelium as does olfactory marker protein (OMP) mRNA. In contrast to the latter, CaM mRNA makes a partial recovery by one month after surgery. These results, coupled with those from in situ hybridization, indicate that CaM mRNA is expressed in both mature and immature olfactory neurons. The program regulating CaM gene expression in olfactory neurons is distinct from those controlling expression of B50/GAP43 in immature, or OMP in mature, neurons respectively.

OpenMP-accelerated SWAT simulation using Intel C and FORTRAN compilers: Development and benchmark

NASA Astrophysics Data System (ADS)

Ki, Seo Jin; Sugimura, Tak; Kim, Albert S.

2015-02-01

We developed a practical method to accelerate execution of Soil and Water Assessment Tool (SWAT) using open (free) computational resources. The SWAT source code (rev 622) was recompiled using a non-commercial Intel FORTRAN compiler in Ubuntu 12.04 LTS Linux platform, and newly named iOMP-SWAT in this study. GNU utilities of make, gprof, and diff were used to develop the iOMP-SWAT package, profile memory usage, and check identicalness of parallel and serial simulations. Among 302 SWAT subroutines, the slowest routines were identified using GNU gprof, and later modified using Open Multiple Processing (OpenMP) library in an 8-core shared memory system. In addition, a C wrapping function was used to rapidly set large arrays to zero by cross compiling with the original SWAT FORTRAN package. A universal speedup ratio of 2.3 was achieved using input data sets of a large number of hydrological response units. As we specifically focus on acceleration of a single SWAT run, the use of iOMP-SWAT for parameter calibrations will significantly improve the performance of SWAT optimization.

Distribution of Chlamydia trachomatis serotypes in clinical urogenital samples from north-eastern Croatia.

PubMed

Bošnjak, Zinka; Džijan, Snježana; Pavlinić, Dinko; Perić, Magdalena; Ružman, Nataša; Križan, Ivana Roksandić; Lauc, Gordan; Antolović-Požgain, Arlen; Burazin, Jelena; Vuković, Dubravka

2012-06-01

The purpose of this study was to determine prevalence of Chlamydia trachomatis (Ct) urogenital infection and its serotype distribution from clinical samples in north-eastern Croatia. During a 3-year period, 2,379 urogenital samples were analyzed by real-time polymerase chain reaction (A group), while 4,846 genital swabs were analyzed by direct fluorescent antibody test (B group). 132 Ct positive specimens were genotyped by omp1 gene sequencing. The prevalence rate of Ct was 3.2 % in A and 1 % in B group. The most prevalent chlamydial genotype was E (44 %), followed by F (33 %), K (11.5 %), G (8 %), J/UW (5.3 %), D-IC (4.4 %), D-B120 (1.8 %), and B/IU, J/IU, Ia/IU (0.9 % each) serotypes. Single-nucleotide polymorphisms (SNPs) of omp1 gene were detected in E, K, and G serotypes. Some of these SNPs (C/T at position 272 and G/A at position 813 in E strain; C/T at position 884 in D strain) might represent novel omp1 variants.

Comparison of potential protection conferred by three immunization strategies (protein/protein, DNA/DNA, and DNA/protein) against Brucella infection using Omp2b in BALB/c Mice.

PubMed

Golshani, Maryam; Rafati, Sima; Nejati-Moheimani, Mehdi; Ghasemian, Melina; Bouzari, Saeid

2016-12-25

In the present study, immunogenicity and protective efficacy of the Brucella outer membrane protein 2b (Omp2b) was evaluated in BALB/c mice using Protein/Protein, DNA/DNA and DNA/Protein vaccine strategies. Immunization of mice with three vaccine regimens elicited a strong specific IgG response (higher IgG2a titers over IgG1 titers) and provided Th1-oriented immune response. Vaccination of BALB/c mice with the DNA/Pro regimen induced higher levels of IFN-γ/IL-2 and conferred more protection levels against B. melitenisis and B. abortus challenge than did the protein or DNA alone. In conclusion, Omp2b is able to stimulate specific immune responses and to confer cross protection against B. melitensis and B. abortus infection. Therefore, it could be introduced as a new potential candidate for the development of a subunit vaccine against Brucella infection. Copyright © 2016 Elsevier B.V. All rights reserved.

Micro-scale NMR Screening of New Detergents for Membrane Protein Structural Biology

PubMed Central

Zhang, Qinghai; Horst, Reto; Geralt, Michael; Ma, Xingquan; Hong, Wen-Xu; Finn, M. G.; Stevens, Raymond C.; Wüthrich, Kurt

2008-01-01

The rate limiting step in biophysical characterization of membrane proteins is often the availability of suitable amounts of protein material. It was therefore of interest to demonstrate that micro-coil nuclear magnetic resonance (NMR) technology can be used to screen microscale quantities of membrane proteins for proper folding in samples destined for structural studies. Micoscale NMR was then used to screen a series of newly designed zwitterionic phosphocholine detergents for their ability to reconstitute membrane proteins, using the previously well characterized β-barrel E.coli outer membrane protein OmpX as a test case. Fold screening was thus achieved with μg-amounts of uniformly 2H,15N-labeld OmpX and affordable amounts of the detergents, and prescreening with SDS-gel electrophoresis ensured efficient selection of the targets for NMR studies. A systematic approach to optimize the phosphocholine motif for membrane protein refolding led to the identification of two new detergents, 138-Fos and 179-Fos, that yield 2D [15N,1H]-TROSY correlation NMR spectra of natively folded reconstituted OmpX. PMID:18479092

Molecular cloning, sequencing, and expression of the outer membrane protein P2 gene of Haemophilus parasuis.

PubMed

Li, Peng; Bai, Juan; Li, Jun-xing; Zhang, Guo-long; Song, Yan-hua; Li, Yu-feng; Wang, Xian-wei; Jiang, Ping

2012-10-01

Haemophilus parasuis is the etiological agent of Glässer's disease characterized by fibrinous polyserositis, polyarthritis, and meningitis in young pigs. But it is difficult to develop universal serological diagnostic tools and effective vaccines against this disease because of the serovar diversity of the isolates. In this study, enterobacterial repetitive intergenic consensus-polymerase chain reaction, were performed to investigate the gene profile of 111 isolates of H. parasuis from China. And a specific common gene of H. parasuis was cloned and identified as the outer-membrane protein (OMP) P2 gene. Sequencing results of OMP P2 genes of 22 isolates showed that they had high homology and could be divided into 2 genetic types. Moreover, the OMPP2 protein was expressed in Escherichia coli expressing system. And the purified recombinant protein provided partial protection against H. parasuis infection in mice. It suggested the OMP P2 was an immunogenic protein and had great potential to serve as a vaccine and diagnostic antigen. Copyright © 2011 Elsevier Ltd. All rights reserved.

Iron restriction and the growth of Salmonella enteritidis.

PubMed Central

Chart, H.; Rowe, B.

1993-01-01

Strains of Salmonella enteritidis were examined for their ability to remove ferric-ions from the iron chelating agents ovotransferrin, Desferal and EDDA. Growth of S. enteritidis phage type (PT) 4 (SE4) in trypticase soy broth containing ovotransferrin resulted in the expression of iron regulated outer membrane proteins (OMPs) of 74, 78 and 81 kDa, and unexpectedly the repression of expression of OMP C. The 38 MDa 'mouse virulence' plasmid was not required for the expression of the iron-regulated OMPs (IROMPs). SE4 was able to obtain iron bound to the iron chelator Desferal and EDDA without expressing a high-affinity iron uptake system. Strains of S. enteritidis belonging to PTs 7, 8, 13a, 23, 24 and 30 were also able to remove ferric ions from Desferal and EDDA without expressing a high-affinity iron uptake system. We conclude that strains of SE4 possess a high-affinity iron sequestering mechanism that can readily remove iron from ovotransferrin. It is likely that iron limitation, and not iron restriction, is responsible for the bacteriostatic properties of fresh egg whites. Images Fig. 2 PMID:8432322

Resolved single-molecule detection of individual species within a mixture of anti-biotin antibodies using an engineered monomeric nanopore.

PubMed

Fahie, Monifa; Chisholm, Christina; Chen, Min

2015-02-24

Oligomeric protein nanopores with rigid structures have been engineered for the purpose of sensing a wide range of analytes including small molecules and biological species such as proteins and DNA. We chose a monomeric β-barrel porin, OmpG, as the platform from which to derive the nanopore sensor. OmpG is decorated with seven flexible loops that move dynamically to create a distinct gating pattern when ionic current passes through the pore. Biotin was chemically tethered to the most flexible one of these loops. The gating characteristic of the loop's movement in and out of the porin was substantially altered by analyte protein binding. The gating characteristics of the pore with bound targets were remarkably sensitive to molecular identity, even providing the ability to distinguish between homologues within an antibody mixture. A total of five gating parameters were analyzed for each analyte to create a unique fingerprint for each biotin-binding protein. Our exploitation of gating noise as a molecular identifier may allow more sophisticated sensor design, while OmpG's monomeric structure greatly simplifies nanopore production.

Escherichia coli K1 invasion increases human brain microvascular endothelial cell monolayer permeability by disassembling vascular-endothelial cadherins at tight junctions.

PubMed

Sukumaran, Sunil K; Prasadarao, Nemani V

2003-11-01

We investigated the permeability changes that occur in the human brain microvascular endothelial cell (HBMEC) monolayer, an in vitro model of the blood-brain barrier, during Escherichia coli K1 infection. An increase in permeability of HBMECs and a decrease in transendothelial electrical resistance were observed. These permeability changes occurred only when HBMECs were infected with E. coli expressing outer membrane protein A (OmpA) and preceded the traversal of bacteria across the monolayer. Activated protein kinase C (PKC)-alpha interacts with vascular-endothelial cadherins (VECs) at the tight junctions of HBMECs, resulting in the dissociation of beta-catenins from VECs and leading to the increased permeability of the HBMEC monolayer. Overexpression of a dominant negative form of PKC-alpha in HBMECs blocked the E. coli-induced increase in permeability of HBMECs. Anti-OmpA and anti-OmpA receptor antibodies exerted inhibition of E. coli-induced permeability of HBMEC monolayers. This inhibition was the result of the absence of PKC-alpha activation in HBMECs treated with the antibodies.

Bandwidth efficient channel estimation method for airborne hyperspectral data transmission in sparse doubly selective communication channels

NASA Astrophysics Data System (ADS)

Vahidi, Vahid; Saberinia, Ebrahim; Regentova, Emma E.

2017-10-01

A channel estimation (CE) method based on compressed sensing (CS) is proposed to estimate the sparse and doubly selective (DS) channel for hyperspectral image transmission from unmanned aircraft vehicles to ground stations. The proposed method contains three steps: (1) the priori estimate of the channel by orthogonal matching pursuit (OMP), (2) calculation of the linear minimum mean square error (LMMSE) estimate of the received pilots given the estimated channel, and (3) estimate of the complex amplitudes and Doppler shifts of the channel using the enhanced received pilot data applying a second round of a CS algorithm. The proposed method is named DS-LMMSE-OMP, and its performance is evaluated by simulating transmission of AVIRIS hyperspectral data via the communication channel and assessing their fidelity for the automated analysis after demodulation. The performance of the DS-LMMSE-OMP approach is compared with that of two other state-of-the-art CE methods. The simulation results exhibit up to 8-dB figure of merit in the bit error rate and 50% improvement in the hyperspectral image classification accuracy.

First Observations of SO2 from the Satellite Suomi NPP OMPS: Widespread Air Pollution Events Over China

NASA Technical Reports Server (NTRS)

Yang, Kai; Dickerson, Russell R.; Carn, Simon A.; Ge, Cui; Wang, Jun

2013-01-01

Severe smog episodes over China in January 2013 received worldwide attention. This air pollution was distinguished by heavy loadings of fine particulate matter and SO2. To characterize these episodes, we employed the Ozone Mapping and Profiler Suite, Nadir Mapper (OMPS NM), an ultraviolet (UV) spectrometer flying on the Suomi National Polar-orbiting Partnership (SNPP) spacecraft since October 2011. We developed an advanced algorithm to quantify SO2 in the lower troposphere and achieved high-quality retrievals from OMPS NM, which are characterized by high precision, approx. 0.2 Dobson Units (DU; 1 DU = 2.69 x 10(exp 16) molecules/sq cm) for instantaneous field of view SO2 data and low biases (within +/-0.2 DU). Here we report SO2 retrievals and UV aerosol index data for these pollution events. The SO2 columns and the areas covered by high pollutant concentrations are quantified; the results reveal for the first time the full extent (an area of approx. 10(exp 6) sq km containing up to 60 kt of SO2) of these episodes.

Program budgeting and marginal analysis: a case study in chronic airflow limitation.

PubMed

Crockett, A; Cranston, J; Moss, J; Scown, P; Mooney, G; Alpers, J

1999-01-01

Program budgeting and marginal analysis is a method of priority-setting in health care. This article describes how this method was applied to the management of a disease-specific group, chronic airflow limitation. A sub-program flow chart clarified the major cost drivers. After assessment of the technical efficiency of the sub-programs and careful and detailed analysis, incremental and decremental wish lists of activities were established. Program budgeting and marginal analysis provides a framework for rational resource allocation. The nurturing of a vigorous program management group, with members representing all participants in the process (including patients/consumers), is the key to a successful outcome.

High-Level Carbapenem Resistance in a Klebsiella pneumoniae Clinical Isolate Is Due to the Combination of blaACT-1 β-Lactamase Production, Porin OmpK35/36 Insertional Inactivation, and Down-Regulation of the Phosphate Transport Porin PhoE

PubMed Central

Kaczmarek, Frank M.; Dib-Hajj, Fadia; Shang, Wenchi; Gootz, Thomas D.

2006-01-01

Clinical isolates of Klebsiella pneumoniae resistant to carbapenems and essentially all other antibiotics (multidrug resistant) are being isolated from some hospitals in New York City with increasing frequency. A highly related pair of K. pneumoniae strains isolated on the same day from one patient in a hospital in New York City were studied for antibiotic resistance. One (KP-2) was resistant to imipenem, meropenem, and sulopenem (MICs of 16 to 32 μg/ml) while the other (KP-1) was susceptible (MIC of 0.5 μg/ml); both contained the blaACT-1, blaSHV-1, and blaTEM-1 β-lactamases. blaACT-1 in both strains was encoded on a large ∼150-kb plasmid. Both isolates contained an identical class 1 integron encoding resistance to aminoglycosides and chloramphenicol. They each had identical insertions in ompK35 and ompK36, resulting in disruption of these key porin genes. The carbapenem-resistant and -susceptible isolates were extensively studied for differences in the structural and regulatory genes for the operons acrRAB, marORAB, romA-ramA, soxRS, micF, micC, phoE, phoBR, rpoS, and hfq. No changes were detected between the isolates except for a significant down-regulation of ompK37, phoB, and phoE in KP-2 as deduced from reverse transcription-PCR analysis of mRNA and polyacrylamide gel electrophoresis separation of outer membrane proteins. Backcross analysis was conducted using the wild-type phoE gene cloned into the vector pGEM under regulation of its native promoter as well as the lacZ promoter following transformation into the resistant KP-2 isolate. The wild-type gene reversed carbapenem resistance only when under control of the heterologous lacZ promoter. In the background of ompK35-ompK36 gene disruption, the up-regulation of phoE in KP-1 apparently compensated for porin loss and conferred carbapenem susceptibility. Down-regulation of phoE in KP-2 may represent the normal state of this gene, or it may have been selected from KP-1 in vivo under antibiotic pressure, generating the carbapenem-resistant clone. This is the first study in the Enterobacteriaceae where expression of the phosphate-regulated PhoE porin has been associated with resistance to antimicrobials. Our results with this pair of Klebsiella clinical isolates highlight the complex and evolving nature of multiple drug resistance in this species. PMID:17005822

High-level carbapenem resistance in a Klebsiella pneumoniae clinical isolate is due to the combination of bla(ACT-1) beta-lactamase production, porin OmpK35/36 insertional inactivation, and down-regulation of the phosphate transport porin phoe.

PubMed

Kaczmarek, Frank M; Dib-Hajj, Fadia; Shang, Wenchi; Gootz, Thomas D

2006-10-01

Clinical isolates of Klebsiella pneumoniae resistant to carbapenems and essentially all other antibiotics (multidrug resistant) are being isolated from some hospitals in New York City with increasing frequency. A highly related pair of K. pneumoniae strains isolated on the same day from one patient in a hospital in New York City were studied for antibiotic resistance. One (KP-2) was resistant to imipenem, meropenem, and sulopenem (MICs of 16 to 32 microg/ml) while the other (KP-1) was susceptible (MIC of 0.5 microg/ml); both contained the bla(ACT-1), bla(SHV-1), and bla(TEM-1) beta-lactamases. bla(ACT-1) in both strains was encoded on a large approximately 150-kb plasmid. Both isolates contained an identical class 1 integron encoding resistance to aminoglycosides and chloramphenicol. They each had identical insertions in ompK35 and ompK36, resulting in disruption of these key porin genes. The carbapenem-resistant and -susceptible isolates were extensively studied for differences in the structural and regulatory genes for the operons acrRAB, marORAB, romA-ramA, soxRS, micF, micC, phoE, phoBR, rpoS, and hfq. No changes were detected between the isolates except for a significant down-regulation of ompK37, phoB, and phoE in KP-2 as deduced from reverse transcription-PCR analysis of mRNA and polyacrylamide gel electrophoresis separation of outer membrane proteins. Backcross analysis was conducted using the wild-type phoE gene cloned into the vector pGEM under regulation of its native promoter as well as the lacZ promoter following transformation into the resistant KP-2 isolate. The wild-type gene reversed carbapenem resistance only when under control of the heterologous lacZ promoter. In the background of ompK35-ompK36 gene disruption, the up-regulation of phoE in KP-1 apparently compensated for porin loss and conferred carbapenem susceptibility. Down-regulation of phoE in KP-2 may represent the normal state of this gene, or it may have been selected from KP-1 in vivo under antibiotic pressure, generating the carbapenem-resistant clone. This is the first study in the Enterobacteriaceae where expression of the phosphate-regulated PhoE porin has been associated with resistance to antimicrobials. Our results with this pair of Klebsiella clinical isolates highlight the complex and evolving nature of multiple drug resistance in this species.

The Joint Polar Satellite System (JPSS) Program's Algorithm Change Process (ACP): Past, Present and Future

NASA Technical Reports Server (NTRS)

Griffin, Ashley

2017-01-01

The Joint Polar Satellite System (JPSS) Program Office is the supporting organization for the Suomi National Polar Orbiting Partnership (S-NPP) and JPSS-1 satellites. S-NPP carries the following sensors: VIIRS, CrIS, ATMS, OMPS, and CERES with instruments that ultimately produce over 25 data products that cover the Earths weather, oceans, and atmosphere. A team of scientists and engineers from all over the United States document, monitor and fix errors in operational software code or documentation with the algorithm change process (ACP) to ensure the success of the S-NPP and JPSS 1 missions by maintaining quality and accuracy of the data products the scientific community relies on. This poster will outline the programs algorithm change process (ACP), identify the various users and scientific applications of our operational data products and highlight changes that have been made to the ACP to accommodate operating system upgrades to the JPSS programs Interface Data Processing Segment (IDPS), so that the program is ready for the transition to the 2017 JPSS-1 satellite mission and beyond.

Identification of Novel Surface-Exposed Proteins of Rickettsia rickettsii by Affinity Purification and Proteomics

PubMed Central

Gong, Wenping; Xiong, Xiaolu; Qi, Yong; Jiao, Jun; Duan, Changsong; Wen, Bohai

2014-01-01

Rickettsia rickettsii, the causative agent of Rocky Mountain spotted fever, is the most pathogenic member among Rickettsia spp. Surface-exposed proteins (SEPs) of R. rickettsii may play important roles in its pathogenesis or immunity. In this study, R. rickettsii organisms were surface-labeled with sulfo-NHS-SS-biotin and the labeled proteins were affinity-purified with streptavidin. The isolated proteins were separated by two-dimensional electrophoresis, and 10 proteins were identified among 23 protein spots by electrospray ionization tandem mass spectrometry. Five (OmpA, OmpB, GroEL, GroES, and a DNA-binding protein) of the 10 proteins were previously characterized as surface proteins of R. rickettsii. Another 5 proteins (Adr1, Adr2, OmpW, Porin_4, and TolC) were first recognized as SEPs of R. rickettsii herein. The genes encoding the 5 novel SEPs were expressed in Escherichia coli cells, resulting in 5 recombinant SEPs (rSEPs), which were used to immunize mice. After challenge with viable R. rickettsii cells, the rickettsial load in the spleen, liver, or lung of mice immunized with rAdr2 and in the lungs of mice immunized with other rSEPs excluding rTolC was significantly lower than in mice that were mock-immunized with PBS. The in vitro neutralization test revealed that sera from mice immunized with rAdr1, rAdr2, or rOmpW reduced R. rickettsii adherence to and invasion of vascular endothelial cells. The immuno-electron microscopic assay clearly showed that the novel SEPs were located in the outer and/or inner membrane of R. rickettsii. Altogether, the 5 novel SEPs identified herein might be involved in the interaction of R. rickettsii with vascular endothelial cells, and all of them except TolC were protective antigens. PMID:24950252

Construction and phase I clinical evaluation of the safety and immunogenicity of a candidate enterotoxigenic Escherichia coli vaccine strain expressing colonization factor antigen CFA/I.

PubMed

Turner, Arthur K; Beavis, Juliet C; Stephens, Jonathan C; Greenwood, Judith; Gewert, Cornelia; Thomas, Nicola; Deary, Alison; Casula, Gabriella; Daley, Alexandra; Kelly, Paul; Randall, Roger; Darsley, Michael J

2006-02-01

Oral delivery of toxin-negative derivatives of enterotoxigenic Escherichia coli (ETEC) that express colonization factor antigens (CFA) with deletions of the aroC, ompC, ompF, and toxin genes may be an effective approach to vaccination against ETEC-associated diarrhea. We describe the creation and characterization of an attenuated CFA/I-expressing ETEC vaccine candidate, ACAM2010, from a virulent isolate in which the heat-stable enterotoxin (ST) and CFA/I genes were closely linked and on the same virulence plasmid as the enteroaggregative E. coli heat-stable toxin (EAST1) gene. A new suicide vector (pJCB12) was constructed and used to delete the ST and EAST1 genes and to introduce defined deletion mutations into the aroC, ompC, and ompF chromosomal genes. A phase I trial, consisting of an open-label dose escalation phase in 18 adult outpatient volunteers followed by a placebo-controlled double-blind phase in an additional 31 volunteers, was conducted. The vaccine was administered in two formulations, fresh culture and frozen suspension. These were both well tolerated, with no evidence of significant adverse events related to vaccination. Immunoglobulin A (IgA) and IgG antibody-secreting cells specific for CFA/I were assayed by ELISPOT. Positive responses (greater than twofold increase) were seen in 27 of 37 (73%) subjects who received the highest dose level of vaccine (nominally 5 x 10(9) CFU). Twenty-nine of these volunteers were secreting culturable vaccine organisms at day 3 following vaccination; five were still positive on day 7, with a single isolation on day 13. This live attenuated bacterial vaccine is safe and immunogenic in healthy adult volunteers.

Leptospiral outer membrane protein microarray, a novel approach to identification of host ligand-binding proteins.

PubMed

Pinne, Marija; Matsunaga, James; Haake, David A

2012-11-01

Leptospirosis is a zoonosis with worldwide distribution caused by pathogenic spirochetes belonging to the genus Leptospira. The leptospiral life cycle involves transmission via freshwater and colonization of the renal tubules of their reservoir hosts. Infection requires adherence to cell surfaces and extracellular matrix components of host tissues. These host-pathogen interactions involve outer membrane proteins (OMPs) expressed on the bacterial surface. In this study, we developed an Leptospira interrogans serovar Copenhageni strain Fiocruz L1-130 OMP microarray containing all predicted lipoproteins and transmembrane OMPs. A total of 401 leptospiral genes or their fragments were transcribed and translated in vitro and printed on nitrocellulose-coated glass slides. We investigated the potential of this protein microarray to screen for interactions between leptospiral OMPs and fibronectin (Fn). This approach resulted in the identification of the recently described fibronectin-binding protein, LIC10258 (MFn8, Lsa66), and 14 novel Fn-binding proteins, denoted Microarray Fn-binding proteins (MFns). We confirmed Fn binding of purified recombinant LIC11612 (MFn1), LIC10714 (MFn2), LIC11051 (MFn6), LIC11436 (MFn7), LIC10258 (MFn8, Lsa66), and LIC10537 (MFn9) by far-Western blot assays. Moreover, we obtained specific antibodies to MFn1, MFn7, MFn8 (Lsa66), and MFn9 and demonstrated that MFn1, MFn7, and MFn9 are expressed and surface exposed under in vitro growth conditions. Further, we demonstrated that MFn1, MFn4 (LIC12631, Sph2), and MFn7 enable leptospires to bind fibronectin when expressed in the saprophyte, Leptospira biflexa. Protein microarrays are valuable tools for high-throughput identification of novel host ligand-binding proteins that have the potential to play key roles in the virulence mechanisms of pathogens.

Identification of OmpR-Family Response Regulators Interacting with Thioredoxin in the Cyanobacterium Synechocystis sp. PCC 6803

PubMed Central

Kadowaki, Taro; Nishiyama, Yoshitaka; Hisabori, Toru; Hihara, Yukako

2015-01-01

The redox state of the photosynthetic electron transport chain is known to act as a signal to regulate the transcription of key genes involved in the acclimation responses to environmental changes. We hypothesized that the protein thioredoxin (Trx) acts as a mediator connecting the redox state of the photosynthetic electron transport chain and transcriptional regulation, and established a screening system to identify transcription factors (TFs) that interact with Trx. His-tagged TFs and S-tagged mutated form of Trx, TrxMC35S, whose active site cysteine 35 was substituted with serine to trap the target interacting protein, were co-expressed in E. coli cells and Trx-TF complexes were detected by immuno-blotting analysis. We examined the interaction between Trx and ten OmpR family TFs encoded in the chromosome of the cyanobacterium Synechocystis sp. PCC 6803 (S.6803). Although there is a highly conserved cysteine residue in the receiver domain of all OmpR family TFs, only three, RpaA (Slr0115), RpaB (Slr0946) and ManR (Slr1837), were identified as putative Trx targets. The recombinant forms of wild-type TrxM, RpaA, RpaB and ManR proteins from S.6803 were purified following over-expression in E. coli and their interaction was further assessed by monitoring changes in the number of cysteine residues with free thiol groups. An increase in the number of free thiols was observed after incubation of the oxidized TFs with Trx, indicating the reduction of cysteine residues as a consequence of interaction with Trx. Our results suggest, for the first time, the possible regulation of OmpR family TFs through the supply of reducing equivalents from Trx, as well as through the phospho-transfer from its cognate sensor histidine kinase. PMID:25774906

The Vibrio parahaemolyticus ToxRS Regulator Is Required for Stress Tolerance and Colonization in a Novel Orogastric Streptomycin-Induced Adult Murine Model

PubMed Central

Whitaker, W. Brian; Parent, Michelle A.; Boyd, Aoife; Richards, Gary P.

2012-01-01

Vibrio parahaemolyticus, a marine bacterium, is the causative agent of gastroenteritis associated with the consumption of seafood. It contains a homologue of the toxRS operon that in V. cholerae is the key regulator of virulence gene expression. We examined a nonpolar mutation in toxRS to determine the role of these genes in V. parahaemolyticus RIMD2210633, an O3:K6 isolate, and showed that compared to the wild type, ΔtoxRS was significantly more sensitive to acid, bile salts, and sodium dodecyl sulfate stresses. We demonstrated that ToxRS is a positive regulator of ompU expression, and that the complementation of ΔtoxRS with ompU restores stress tolerance. Furthermore, we showed that ToxRS also regulates type III secretion system genes in chromosome I via the regulation of the leuO homologue VP0350. We examined the effect of ΔtoxRS in vivo using a new orogastric adult murine model of colonization. We demonstrated that streptomycin-treated adult C57BL/6 mice experienced prolonged intestinal colonization along the entire intestinal tract by the streptomycin-resistant V. parahaemolyticus. In contrast, no colonization occurred in non-streptomycin-treated mice. A competition assay between the ΔtoxRS and wild-type V. parahaemolyticus strains marked with the β-galactosidase gene lacZ demonstrated that the ΔtoxRS strain was defective in colonization compared to the wild-type strain. This defect was rescued by ectopically expressing ompU. Thus, the defect in stress tolerance and colonization in ΔtoxRS is solely due to OmpU. To our knowledge, the orogastric adult murine model reported here is the first showing sustained intestinal colonization by V. parahaemolyticus. PMID:22392925

The nature of information, required for export and sorting, present within the outer membrane protein OmpA of Escherichia coli K-12.

PubMed

Freudl, R; Schwarz, H; Klose, M; Movva, N R; Henning, U

1985-12-16

Information, in addition to that provided by signal sequences, for translocation across the plasma membrane is thought to be present in exported proteins of Escherichia coli. Such information must also exist for the localization of such proteins. To determine the nature of this information, overlapping inframe deletions have been constructed in the ompA gene which codes for a 325-residue major outer membrane protein. In addition, one deletion, encoding only the NH2-terminal part of the protein up to residue 160, was prepared. The location of each product was determined by immunoelectron microscopy. Proteins missing residues 4-45, 43-84, 46-227, 86-227 or 160-325 of the mature protein were all efficiently translocated across the plasma membrane. The first two proteins were found in the outer membrane, the others in the periplasmic space. It has been proposed that export and sorting signals consist of relatively small amino acid sequences near the NH2 terminus of an outer membrane protein. On the basis of sequence homologies it has also been suggested that such proteins possess a common sorting signal. The locations of the partially deleted proteins described here show that a unique export signal does not exist in the OmpA protein. The proposed common sorting signal spans residues 1-14 of OmpA. Since this region is not essential for routing the protein, the existence of a common sorting signal is doubtful. It is suggested that information both for export (if existent) and localization lies within protein conformation which for the former process should be present repeatedly in the polypeptide.

Contribution of Efflux Pumps, Porins, and β-Lactamases to Multidrug Resistance in Clinical Isolates of Acinetobacter baumannii

PubMed Central

Rumbo, C.; Gato, E.; López, M.; Ruiz de Alegría, C.; Fernández-Cuenca, F.; Martínez-Martínez, L.; Vila, J.; Pachón, J.; Cisneros, J. M.; Rodríguez-Baño, J.; Pascual, A.

2013-01-01

We investigated the mechanisms of resistance to carbapenems, aminoglycosides, glycylcyclines, tetracyclines, and quinolones in 90 multiresistant clinical strains of Acinetobacter baumannii isolated from two genetically unrelated A. baumannii clones: clone PFGE-ROC-1 (53 strains producing the OXA-58 β-lactamase enzyme and 18 strains with the OXA-24 β-lactamase) and clone PFGE-HUI-1 (19 strains susceptible to carbapenems). We used real-time reverse transcriptase PCR to correlate antimicrobial resistance (MICs) with expression of genes encoding chromosomal β-lactamases (AmpC and OXA-51), porins (OmpA, CarO, Omp33, Dcap-like, OprB, Omp25, OprC, OprD, and OmpW), and proteins integral to six efflux systems (AdeABC, AdeIJK, AdeFGH, CraA, AbeM, and AmvA). Overexpression of the AdeABC system (level of expression relative to that by A. baumannii ATCC 17978, 30- to 45-fold) was significantly associated with resistance to tigecycline, minocycline, and gentamicin and other biological functions. However, hyperexpression of the AdeIJK efflux pump (level of expression relative to that by A. baumannii ATCC 17978, 8- to 10-fold) was significantly associated only with resistance to tigecycline and minocycline (to which the TetB efflux system also contributed). TetB and TetA(39) efflux pumps were detected in clinical strains and were associated with resistance to tetracyclines and doxycycline. The absence of the AdeABC system and the lack of expression of other mechanisms suggest that tigecycline-resistant strains of the PFGE-HUI-1 clone may be associated with a novel resistance-nodulation-cell efflux pump (decreased MICs in the presence of the inhibitor Phe-Arg β-naphthylamide dihydrochloride) and the TetA(39) system. PMID:23939894

Presumable role of outer membrane proteins of Salmonella containing sialylated lipopolysaccharides serovar Ngozi, sv. Isaszeg and subspecies arizonae in determining susceptibility to human serum.

PubMed

Futoma-Kołoch, Bożena; Godlewska, Urszula; Guz-Regner, Katarzyna; Dorotkiewicz-Jach, Agata; Klausa, Elżbieta; Rybka, Jacek; Bugla-Płoskońska, Gabriela

2015-01-01

The O48 group comprises Salmonella bacteria containing sialic acid in the lipopolysaccharide (LPS). Bacteria with sialylated surface structures are described as pathogens that avoid immunological response of the host by making similar their surface antigens to the host's tissues (molecular mimicry). It is known that the smooth-type LPS of Salmonella enterica and outer membrane proteins (OMP) PgtE, PagC and Rck mediate serum resistant phenotype by affecting complement system (C). The aim of this study was to investigate C3 component activation by Salmonella O48 LPS and OMP. In the present study, we examined C3 component deposition on the three Salmonella O48 strains: S. enterica subspecies enterica serovar Ngozi, S. enterica subsp. enterica sv. Isaszeg, and S. enterica subsp. arizonae containing sialic acid in the O-specific part of LPS. The greatest C3 deposition occurred on Salmonella sv. Isaszeg cells (p < 0.005) as well as on their LPS (low content of sialic acid in LPS) (p < 0.05) after 45 min of incubation in 50% human serum. Weaker C3 deposition ratio on the Salmonella sv. Ngozi (high content of sialic acid in LPS) and Salmonella subsp. arizonae (high content of sialic acid in LPS) cells correlated with the lower C3 activation on their LPS. Immunoblotting revealed that OMP isolated from the tested strains also bound C3 protein fragments. We suggest that activation of C3 serum protein is dependent on the sialic acid contents in the LPS as well as on the presence of OMP in the range of molecular masses of 35-48 kDa.

Extending the SBUV MOD Ozone Profile data record with OMPS Nadir Profiler Data: Updated Trends and Uncertainties

NASA Astrophysics Data System (ADS)

Frith, S. M.; Stolarski, R. S.; McPeters, R. D.; Kramarova, N. A.

2017-12-01

The Ozone Monitoring and Profile Suite (OMPS) on the Suomi NPP satellite comprises three instruments measuring profile and total column ozone. The Nadir Profiler sensor measures broadly-resolved vertical ozone profiles retrieved from backscattered UV radiances, and continues a nearly unbroken record of measurements from the Solar Backscatter Ultraviolet (SBUV and SBUV/2) series of instruments dating back to late 1978. The SBUV Merged Ozone Dataset (MOD) combines data from the SBUV instrument series into a single coherent data record. The last instrument in the series, operating on the NOAA 19 satellite, is expected to encounter higher measurement uncertainties as the N19 orbit drifts closer to the terminator, necessitating a move to the next generation OMPS instruments. Here we incorporate OMPS NP v2.3 data from 2012-2017 into the MOD record and evaluate the effects of the new data on theoverall record, particularly the sensitivity of long-term trend estimates derived from MOD. We will evaluate the uncertainty associated with merging multiple records. We use a Monte Carlo modeling approach to estimate the potential for uncertainties in the calibration and drift of individual instruments to mimic long-term variations in the merged data set. Intra-instrument comparisons during overlap periods are used to quantify the uncertainty of each instrument in the Monte Carlo simulations. Current error estimates using this approach are likely conservative because we model a Gaussian distribution of potential offsets and drifts when the actual distributions are more complicated. In this work we will investigate the effects of the additional data set, but also pursue approaches to define the Monte Carlo model more precisely to better characterize the potential error.

A Negative Regulator of Cellulose Biosynthesis, bcsR, Affects Biofilm Formation, and Adhesion/Invasion Ability of Cronobacter sakazakii.

PubMed

Gao, Jian-Xin; Li, Ping; Du, Xin-Jun; Han, Zhong-Hui; Xue, Rui; Liang, Bin; Wang, Shuo

2017-01-01

Cronobacter sakazakii is an important foodborne pathogen that causes neonatal meningitis and sepsis, with high mortality in neonates. However, very little information is available regarding the pathogenesis of C. sakazakii at the genetic level. In our previous study, a cellulose biosynthesis-related gene ( bcsR ) was shown to be involved in C. sakazakii adhesion/invasion into epithelial cells. In this study, the detailed functions of this gene were investigated using a gene knockout technique. A bcsR knockout mutant (Δ bcsR ) of C. sakazakii ATCC BAA-894 showed decreased adhesion/invasion (3.9-fold) in human epithelial cell line HCT-8. Biofilm formation by the mutant was reduced to 50% of that exhibited by the wild-type (WT) strain. Raman spectrometry was used to detect variations in biofilm components caused by bcsR knockout, and certain components, including carotenoids, fatty acids, and amides, were significantly reduced. However, another biofilm component, cellulose, was increased in Δ bcsR , suggesting that bcsR negatively affects cellulose biosynthesis. This result was also verified via RT-PCR, which demonstrated up-regulation of five crucial cellulose synthesis genes ( bcsA, B, C, E, Q ) in Δ bcsR . Furthermore, the expression of other virulence or biofilm-related genes, including flagellar assembly genes ( fliA, C, D ) and toxicity-related genes ( ompA, ompX, hfq ), was studied. The expression of fliC and ompA in the Δ bcsR mutant was found to be remarkably reduced compared with that in the wild-type and the others were also affected excepted ompX . In summary, bcsR is a negative regulator of cellulose biosynthesis but positively regulates biofilm formation and the adhesion/invasion ability of C. sakazakii .

The Interaction of N-Glycans in Fcγ Receptor I α-Chain with Escherichia coli K1 Outer Membrane Protein A for Entry into Macrophages

PubMed Central

Krishnan, Subramanian; Liu, Fan; Abrol, Ravinder; Hodges, Jacqueline; Goddard, William A.; Prasadarao, Nemani V.

2014-01-01

Neonatal meningitis, caused by Escherichia coli K1, is a serious central nervous system disease. We have established that macrophages serve as permissive niches for E. coli K1 to multiply in the host and for attaining a threshold level of bacterial load, which is a prerequisite for the onset of the disease. Here, we demonstrate experimentally that three N-glycans in FcγRIa interact with OmpA of E. coli K1 for binding to and entering the macrophages. Adoptive transfer of FcγRIa−/− bone marrow-derived macrophages transfected with FcγRIa into FcγRIa−/− newborn mice renders them susceptible to E. coli K1-induced meningitis. In contrast, mice that received bone marrow-derived macrophages transfected with FcγRIa in which N-glycosylation sites 1, 4, and 5 are mutated to alanines exhibit resistance to E. coli K1 infection. Our molecular dynamics and simulation studies predict that N-glycan 5 exhibits strong binding at the barrel site of OmpA formed by loops 3 and 4, whereas N-glycans 1 and 4 interact with loops 1, 3, and 4 of OmpA at tip regions. Molecular modeling data also suggest no role for the IgG binding site in the invasion process. In agreement, experimental mutations in IgG binding site had no effect on the E. coli K1 entry into macrophages in vitro or on the onset of meningitis in newborn mice. Together, this integration of experimental and computational studies reveals how the N-glycans in FcγRIa interact with the OmpA of E. coli K1 for inducing the disease pathogenesis. PMID:25231998

Enzyme architecture: deconstruction of the enzyme-activating phosphodianion interactions of orotidine 5'-monophosphate decarboxylase.

PubMed

Goldman, Lawrence M; Amyes, Tina L; Goryanova, Bogdana; Gerlt, John A; Richard, John P

2014-07-16

The mechanism for activation of orotidine 5'-monophosphate decarboxylase (OMPDC) by interactions of side chains from Gln215 and Try217 at a gripper loop and R235, adjacent to this loop, with the phosphodianion of OMP was probed by determining the kinetic parameters k(cat) and K(m) for all combinations of single, double, and triple Q215A, Y217F, and R235A mutations. The 12 kcal/mol intrinsic binding energy of the phosphodianion is shown to be equal to the sum of the binding energies of the side chains of R235 (6 kcal/mol), Q215 (2 kcal/mol), Y217 (2 kcal/mol), and hydrogen bonds to the G234 and R235 backbone amides (2 kcal/mol). Analysis of a triple mutant cube shows small (ca. 1 kcal/mol) interactions between phosphodianion gripper side chains, which are consistent with steric crowding of the side chains around the phosphodianion at wild-type OMPDC. These mutations result in the same change in the activation barrier to the OMPDC-catalyzed reactions of the whole substrate OMP and the substrate pieces (1-β-D-erythrofuranosyl)orotic acid (EO) and phosphite dianion. This shows that the transition states for these reactions are stabilized by similar interactions with the protein catalyst. The 12 kcal/mol intrinsic phosphodianion binding energy of OMP is divided between the 8 kcal/mol of binding energy, which is utilized to drive a thermodynamically unfavorable conformational change of the free enzyme, resulting in an increase in (k(cat))(obs) for OMPDC-catalyzed decarboxylation of OMP, and the 4 kcal/mol of binding energy, which is utilized to stabilize the Michaelis complex, resulting in a decrease in (K(m))(obs).

Rapid Induction of High-Level Carbapenem Resistance in Heteroresistant KPC-Producing Klebsiella pneumoniae

PubMed Central

Adams-Sapper, Sheila; Nolen, Shantell; Donzelli, Grace Fox; Lal, Mallika; Chen, Kunihiko; Justo da Silva, Livia Helena; Moreira, Beatriz M.

2015-01-01

Enterobacteriaceae strains producing the Klebsiella pneumoniae carbapenemase (KPC) have disseminated worldwide, causing an urgent threat to public health. KPC-producing strains often exhibit low-level carbapenem resistance, which may be missed by automated clinical detection systems. In this study, eight Klebsiella pneumoniae strains with heterogeneous resistance to imipenem were used to elucidate the factors leading from imipenem susceptibility to high-level resistance as defined by clinical laboratory testing standards. Time-kill analysis with an inoculum as low as 3 × 106 CFU/ml and concentrations of imipenem 8- and 16-fold higher than the MIC resulted in the initial killing of 99.9% of the population. However, full recovery of the population occurred by 20 h of incubation in the same drug concentrations. Population profiles showed that recovery was mediated by a heteroresistant subpopulation at a frequency of 2 × 10−7 to 3 × 10−6. Samples selected 2 h after exposure to imipenem were as susceptible as the unexposed parental strain and produced the major outer membrane porin OmpK36. However, between 4 to 8 h after exposure, OmpK36 became absent, and the imipenem MIC increased at least 32-fold. Individual colonies isolated from cultures after 20 h of exposure revealed both susceptible and resistant subpopulations. Once induced, however, the high-level imipenem resistance was maintained, and OmpK36 remained unexpressed even without continued carbapenem exposure. This study demonstrates the essential coordination between blaKPC and ompK36 expression mediating high-level imipenem resistance from a population of bacteria that initially exhibits a carbapenem-susceptibility phenotype. PMID:25801565

TprC/D (Tp0117/131), a trimeric, pore-forming rare outer membrane protein of Treponema pallidum, has a bipartite domain structure.

PubMed

Anand, Arvind; Luthra, Amit; Dunham-Ems, Star; Caimano, Melissa J; Karanian, Carson; LeDoyt, Morgan; Cruz, Adriana R; Salazar, Juan C; Radolf, Justin D

2012-05-01

Identification of Treponema pallidum rare outer membrane proteins (OMPs) has been a longstanding objective of syphilis researchers. We recently developed a consensus computational framework that employs a battery of cellular localization and topological prediction tools to generate ranked clusters of candidate rare OMPs (D. L. Cox et al., Infect. Immun. 78:5178-5194, 2010). TP0117/TP0131 (TprC/D), a member of the T. pallidum repeat (Tpr) family, was a highly ranked candidate. Circular dichroism, heat modifiability by SDS-PAGE, Triton X-114 phase partitioning, and liposome incorporation confirmed that full-length, recombinant TprC (TprC(Fl)) forms a β-barrel capable of integrating into lipid bilayers. Moreover, TprC(Fl) increased efflux of terbium-dipicolinic acid complex from large unilamellar vesicles and migrated as a trimer by blue-native PAGE. We found that in T. pallidum, TprC is heat modifiable, trimeric, expressed in low abundance, and, based on proteinase K accessibility and opsonophagocytosis assays, surface exposed. From these collective data, we conclude that TprC is a bona fide rare OMP as well as a functional ortholog of Escherichia coli OmpF. We also discovered that TprC has a bipartite architecture consisting of a soluble N-terminal portion (TprC(N)), presumably periplasmic and bound directly or indirectly to peptidoglycan, and a C-terminal β-barrel (TprC(C)). Syphilitic rabbits generate antibodies exclusively against TprC(C), while secondary syphilis patients fail to mount a detectable antibody response against either domain. The syphilis spirochete appears to have resolved a fundamental dilemma arising from its extracellular lifestyle, namely, how to enhance OM permeability without increasing its vulnerability to the antibody-mediated defenses of its natural human host.

Entropy-enthalpy compensation at the single protein level: pH sensing in the bacterial channel OmpF.

PubMed

Alcaraz, Antonio; Queralt-Martín, María; Verdiá-Báguena, Carmina; Aguilella, Vicente M; Mafé, Salvador

2014-12-21

The pH sensing mechanism of the OmpF channel operates via ligand modification: increasing acidity induces the replacement of cations with protons in critical binding sites decreasing the channel conductance. Aside from the change in enthalpy associated with the binding, there is also a change in the microscopic arrangements of ligands, receptors and the surrounding solvent. We show that the pH-modulation of the single channel conduction involves small free energy changes because large enthalpic and entropic contributions change in opposite ways, demonstrating an approximate enthalpy-entropy compensation for different salts and concentrations.

Mismatch and resolution in compressive imaging

NASA Astrophysics Data System (ADS)

Fannjiang, Albert; Liao, Wenjing

2011-09-01

Highly coherent sensing matrices arise in discretization of continuum problems such as radar and medical imaging when the grid spacing is below the Rayleigh threshold as well as in using highly coherent, redundant dictionaries as sparsifying operators. Algorithms (BOMP, BLOOMP) based on techniques of band exclusion and local optimization are proposed to enhance Orthogonal Matching Pursuit (OMP) and deal with such coherent sensing matrices. BOMP and BLOOMP have provably performance guarantee of reconstructing sparse, widely separated objects independent of the redundancy and have a sparsity constraint and computational cost similar to OMP's. Numerical study demonstrates the effectiveness of BLOOMP for compressed sensing with highly coherent, redundant sensing matrices.

A New Algorithm for Detecting Cloud Height using OMPS/LP Measurements

NASA Technical Reports Server (NTRS)

Chen, Zhong; DeLand, Matthew; Bhartia, Pawan K.

2016-01-01

The Ozone Mapping and Profiler Suite Limb Profiler (OMPS/LP) ozone product requires the determination of cloud height for each event to establish the lower boundary of the profile for the retrieval algorithm. We have created a revised cloud detection algorithm for LP measurements that uses the spectral dependence of the vertical gradient in radiance between two wavelengths in the visible and near-IR spectral regions. This approach provides better discrimination between clouds and aerosols than results obtained using a single wavelength. Observed LP cloud height values show good agreement with coincident Cloud-Aerosol Lidar and Infrared Pathfinder Satellite Observation (CALIPSO) measurements.

Reconstruction of hyperspectral image using matting model for classification

NASA Astrophysics Data System (ADS)

Xie, Weiying; Li, Yunsong; Ge, Chiru

2016-05-01

Although hyperspectral images (HSIs) captured by satellites provide much information in spectral regions, some bands are redundant or have large amounts of noise, which are not suitable for image analysis. To address this problem, we introduce a method for reconstructing the HSI with noise reduction and contrast enhancement using a matting model for the first time. The matting model refers to each spectral band of an HSI that can be decomposed into three components, i.e., alpha channel, spectral foreground, and spectral background. First, one spectral band of an HSI with more refined information than most other bands is selected, and is referred to as an alpha channel of the HSI to estimate the hyperspectral foreground and hyperspectral background. Finally, a combination operation is applied to reconstruct the HSI. In addition, the support vector machine (SVM) classifier and three sparsity-based classifiers, i.e., orthogonal matching pursuit (OMP), simultaneous OMP, and OMP based on first-order neighborhood system weighted classifiers, are utilized on the reconstructed HSI and the original HSI to verify the effectiveness of the proposed method. Specifically, using the reconstructed HSI, the average accuracy of the SVM classifier can be improved by as much as 19%.

Identification of outer membrane proteins with emulsifying activity by prediction of beta-barrel regions.

PubMed

Walzer, Gil; Rosenberg, Eugene; Ron, Eliora Z

2009-01-01

Microbial bioemulsifiers are secreted by many bacteria and are important for bacterial interactions with hydrophobic substrates or nutrients and for a variety of biotechnological applications. We have recently shown that the OmpA protein in several members of the Acinetobacter family has emulsifying properties. These properties of OmpA depend on the amino acid composition of four putative extra-membrane loops, which in various strains of Acinetobacter, but not in E. coli, are highly hydrophobic. As many Acinetobacter strains can utilize hydrophobic carbon sources, such as oil, the emulsifying activity of their OmpA may be important for the utilization and uptake of hydrocarbons. We assumed that if outer membrane proteins with emulsifying activity are physiologically important, they may exist in additional oil degrading bacteria. In order to identify such proteins, it was necessary to obtain bioinformatics-based predictions for hydrophobic extra-membrane loops. Here we describe a method for using protein sequence data for predicting the hydrophobic properties of the extra-membrane loops of outer membrane proteins. The feasibility of this method is demonstrated by its use to identify a new microbial bioemulsifier - OprG - an outer membrane protein of the oil degrading Pseudomonas putida KT2440.

Selection of organic process and source indicator substances for the anthropogenically influenced water cycle.

PubMed

Jekel, Martin; Dott, Wolfgang; Bergmann, Axel; Dünnbier, Uwe; Gnirß, Regina; Haist-Gulde, Brigitte; Hamscher, Gerd; Letzel, Marion; Licha, Tobias; Lyko, Sven; Miehe, Ulf; Sacher, Frank; Scheurer, Marco; Schmidt, Carsten K; Reemtsma, Thorsten; Ruhl, Aki Sebastian

2015-04-01

An increasing number of organic micropollutants (OMP) is detected in anthropogenically influenced water cycles. Source control and effective natural and technical barriers are essential to maintain a high quality of drinking water resources under these circumstances. Based on the literature and our own research this study proposes a limited number of OMP that can serve as indicator substances for the major sources of OMP, such as wastewater treatment plants, agriculture and surface runoff. Furthermore functional indicators are proposed that allow assessment of the proper function of natural and technical barriers in the aquatic environment, namely conventional municipal wastewater treatment, advanced treatment (ozonation, activated carbon), bank filtration and soil aquifer treatment as well as self-purification in surface water. These indicator substances include the artificial sweetener acesulfame, the anti-inflammatory drug ibuprofen, the anticonvulsant carbamazepine, the corrosion inhibitor benzotriazole and the herbicide mecoprop among others. The chemical indicator substances are intended to support comparisons between watersheds and technical and natural processes independent of specific water cycles and to reduce efforts and costs of chemical analyses without losing essential information. Copyright © 2014 Elsevier Ltd. All rights reserved.

OMPS TC EDR Algorithm: Improvement and Verification

NASA Astrophysics Data System (ADS)

Novicki, M.; Sen, B.; Hao, X.; Qu, J. J.

2009-12-01

The Ozone Mapper and Profiler Suite (OMPS) is scheduled to be launched on the NPOESS Preparatory Project (NPP) platform in early 2011. The OMPS will continue monitoring ozone from space, using three instruments, namely the Total Column Mapper (heritage: TOMS), the Nadir Profiler (heritage: SBUV) and the Limb Profiler (heritage: SOLSE/LORE). The Total Column Mapper (TC) sensor images the Earth through a slit, nadir-cell horizontally spaced at 49.5 km cross-track with an along-track reporting interval of 50 km. The total field of view (FOV) cross track is 110 degrees to provide daily global coverage. The TC sensor, a grating spectrometer, provides 0.45 nm spectral sampling across the wavelength range of 300-380 nm. The calibration stability, which is essential to enable long-term ozone monitoring, is maintained by periodic observations of the Sun, using a diffuser to redirect the solar irradiance into the sensor. We describe the data analysis method being presently implemented to retrieve the total column ozone Earth Data Record (EDR) from the radiance data measured by the TC sensor. We discuss the software changes, the test data used to verify the functional performance and the test results.

GD3/proteosome vaccines induce consistent IgM antibodies against the ganglioside GD3.

PubMed

Livingston, P O; Calves, M J; Helling, F; Zollinger, W D; Blake, M S; Lowell, G H

1993-09-01

The gangliosides of melanoma and other tumours of neuroectodermal origin are suitable targets for immune intervention with tumour vaccines. The optimal vaccines in current use contain ganglioside plus bacillus Calmette-Guérin and induce considerable morbidity. We have screened a variety of new adjuvants in the mouse, and describe one antigen-delivery system, proteosomes, which is especially effective. Highly hydrophobic Neisserial outer membrane proteins (OMP) form multimolecular liposome-like vesicular structures termed proteosomes which can readily incorporate amphiphilic molecules such as GD3 ganglioside. The optimal GD3/proteosome vaccine formulation for induction of GD3 antibodies in the mouse is determined. Interestingly, the use of potent immunological adjuvants in addition to proteosomes augments the IgM and IgG antibody titres against OMP in these vaccines but GD3 antibody titres are unaffected. The application of proteosomes to enhance the immune response to GD3 extends the concept of the proteosome immunopotentiating system from lipopeptides to amphipathic carbohydrate epitopes such as cell-surface gangliosides. The demonstrated safety of meningococcal OMP in humans and the data in mice presented here suggest that proteosome vaccines have potential for augmenting the immunogenicity of amphipathic tumour antigens in humans.

Real-time Volcanic Cloud Products and Predictions for Aviation Alerts

NASA Astrophysics Data System (ADS)

Krotkov, N. A.; Hughes, E. J.; da Silva, A. M., Jr.; Seftor, C. J.; Brentzel, K. W.; Hassinen, S.; Heinrichs, T. A.; Schneider, D. J.; Hoffman, R.; Myers, T.; Flynn, L. E.; Niu, J.; Theys, N.; Brenot, H. H.

2016-12-01

We will discuss progress of the NASA ASP project, which promotes the use of satellite volcanic SO2 (VSO2) and Ash (VA) data, and forecasting tools that enhance VA Decision Support Systems (DSS) at the VA Advisory Centers (VAACs) for prompt aviation warnings. The goals are: (1) transition NASA algorithms to NOAA for global NRT processing and integration into DSS at Washington VAAC for operational users and public dissemination; (2) Utilize Direct Broadcast capability of the Aura and SNPP satellites to process Direct Readout (DR) data at two high latitude locations in Finland and Fairbanks, Alaska to enhance VA DSS in Europe and at USGS's Alaska Volcano Observatory (AVO) and Alaska-VAAC; (3) Improve global Eulerian model-based VA/VSO2 forecasting and risk/cost assessments with Metron Aviation. Our global NRT OMI and OMPS data have been fully integrated into European Support to Aviation Control Service and NOAA operational web sites. We are transitioning OMPS processing to our partners at NOAA/NESDIS to integrate into operational processing environment. NASA's Suomi NPP Ozone Science Team, in conjunction with GSFC's Direct Readout Laboratory (DRL), have implemented Version 2 of the OMPS real-time DR processing package to generate VSO2 and VA products at the Geographic Information Network of Alaska (GINA) and the Finnish Meteorological Institute (FMI). The system provides real-time coverage over some of the most congested airspace and over many of the most active volcanoes in the world. The OMPS real time capability is now publicly available via DRL's IPOPP package. We use satellite observations to define volcanic source term estimates in the NASA GOES-5 model, which was updated allowing for the simulation of VA and VSO2 clouds. Column SO2 observations from SNPP/OMPS provide an initial estimate of the total cloud SO2 mass, and are used with backward transport analysis to make an initial cloud height estimate. Later VSO2 observations are used to "nudge" the SO2 mass within the model. The GEOS-5 simulations provide qualitative forecasts, which locate the extent of regions hazardous to aviation. Air traffic flow algorithms have been developed by Metron Aviation to use GEOS-5 volcanic simulations to determine the most cost-effective rerouting paths around hazardous volcanic clouds.

The effects of IL-1β, IL-8, G-CSF and TNF-α as molecular adjuvant on the immune response to an E. tarda subunit vaccine in flounder (Paralichthys olivaceus).

PubMed

Guo, Ming; Tang, Xiaoqian; Sheng, Xiuzhen; Xing, Jing; Zhan, Wenbin

2018-06-01

Cytokines play vital roles in mounting immune responses and activating host defense network. In this study, the expression plasmid pcDNA3.1 (pcN3) encoding four flounder (Paralichthys olivaceus) cytokines including IL-1β, TNF-α, IL-8 or G-CSF (pcIL-1β, pcTNF-α, pcIL-8 and pcG-CSF) were successfully constructed, and their adjuvant potential on an Edwardsiella tarda (E. tarda) subunit vaccine OmpV (rOmpV) were comparatively analyzed in vaccinated flounder model. Results revealed that flounder vaccinated with rOmpV plus pcIL-1β, pcIL-8 or pcG-CSF produced the relative percent survivals (RPS) of 71%, 65% and 49% respectively, which were higher than that in flounder vaccinated with rOmpV plus pcTNF-α (39%) or pcN3 (36%, the control group). Immunological analysis showed that: (1) except pcTNF-α, higher levels of anti-E. tarda serum antibodies and sIg + lymphocytes in spleen, head kidney and peripheral blood were significantly enhanced by pcIL-1β, pcIL-8 or pcG-CSF, however, pcIL-8 and pcIL-1β enhanced higher levels of sIg + lymphocytes and anti-E. tarda antibodies than pcG-CSF; (2) pcTNF-α could promote the up-regulation of genes participated in cellular immunity (MHCIα, IFN-γ, CD8α and CD8β), pcIL-1β could enhance the expression of genes related to humoral immunity (CD4-1, CD4-2, MHCIIα and IgM), and all the detected genes were augmented by pcIL-8 and pcG-CSF; Among the four cytokines, pcIL-8 and pcIL-1β could strengthen the highest levels of genes participated in cellular immunity and humoral immunity, respectively. These results demonstrated that pcIL-8 and pcIL-1β could enhance stronger cellular and/or humoral immunity induced by rOmpV than pcG-CSF and pcTNF-α, and evoked higher RPS against E. tarda challenge in flounder, which indicated that pcIL-8 and pcIL-1β are promising adjuvants of vaccines in controlling E. tarda infection. Copyright © 2018 Elsevier Ltd. All rights reserved.

Retrieval of ozone profiles from OMPS limb scattering observations

NASA Astrophysics Data System (ADS)

Arosio, Carlo; Rozanov, Alexei; Malinina, Elizaveta; Eichmann, Kai-Uwe; von Clarmann, Thomas; Burrows, John P.

2018-04-01

This study describes a retrieval algorithm developed at the University of Bremen to obtain vertical profiles of ozone from limb observations performed by the Ozone Mapper and Profiler Suite (OMPS). This algorithm is based on the technique originally developed for use with data from the SCanning Imaging Absorption spectroMeter for Atmospheric CHartographY (SCIAMACHY) instrument. As both instruments make limb measurements of the scattered solar radiation in the ultraviolet (UV) and visible (Vis) spectral ranges, an underlying objective of the study is to obtain consolidated and consistent ozone profiles from the two satellites and to produce a combined data set. The retrieval algorithm uses radiances in the UV and Vis wavelength ranges normalized to the radiance at an upper tangent height to obtain ozone concentrations in the altitude range of 12-60 km. Measurements at altitudes contaminated by clouds in the instrument field of view are identified and filtered out. An independent aerosol retrieval is performed beforehand and its results are used to account for the stratospheric aerosol load in the ozone inversion. The typical vertical resolution of the retrieved profiles varies from ˜ 2.5 km at lower altitudes ( < 30 km) to ˜ 1.5 km (about 45 km) and becomes coarser at upper altitudes. The retrieval errors resulting from the measurement noise are estimated to be 1-4 % above 25 km, increasing to 10-30 % in the upper troposphere. OMPS data are processed for the whole of 2016. The results are compared with the NASA product and validated against profiles derived from passive satellite observations or measured in situ by balloon-borne sondes. Between 20 and 60 km, OMPS ozone profiles typically agree with data from the Microwave Limb Sounder (MLS) v4.2 within 5-10 %, whereas in the lower altitude range the bias becomes larger, especially in the tropics. The comparison of OMPS profiles with ozonesonde measurements shows differences within ±5 % between 13 and 30 km at northern middle and high latitudes. At southern middle and high latitudes, an agreement within 5-7 % is also achieved in the same altitude range. An unexpected bias of approximately 10-20 % is detected in the lower tropical stratosphere. The processing of the 2013 data set using the same retrieval settings and its validation against ozonesondes reveals a much smaller bias; a possible reason for this behaviour is discussed.

Validation of OMPS Ozone Profile Data with Expanded Dataset from Brewer and Automated Dobson Network.

NASA Astrophysics Data System (ADS)

Petropavlovskikh, I.; Weatherhead, E.; Cede, A.; Oltmans, S. J.; Kireev, S.; Maillard, E.; Bhartia, P. K.; Flynn, L. E.

2005-12-01

The first NPOESS satellite is scheduled to be launched in 2010 and will carry the Ozone Mapping and Profiler Suite (OMPS) instruments for ozone monitoring. Prior this, the OMPS instruments and algorithms will be tested by flight on the NPOESS/NPP satellite, scheduled for launch in 2008. Pre-launch planning for validation, post launch data validation and verification of the nadir and limb profile algorithm are key components for insuring that the NPOESS will produce a high quality, reliable ozone profile data set. The heritage of satellite instrument validation (TOMS, SBUV, GOME, SCIAMACHY, SAGE, HALOE, ATMOS, etc) has always relied upon surface-based observations. While the global coverage of satellite observations is appealing for validating another satellite, there is no substitute for the hard reference point of a ground-based system such as the Dobson or Brewer network, whose instruments are routinely calibrated and intercompared to standard references. The standard solar occultation instruments, SAGE II and HALOE are well beyond their planned lifetimes and might be inoperative during the OMPS period. The Umkehr network has been one of the key data sets for stratospheric ozone trend calculations and has earned its place as a benchmark network for stratospheric ozone profile observations. The normalization of measurements at different solar zenith angle (SZAs) to the measurement at the smallest SZA cancels out many calibration parameters, including the extra-terrestrial solar flux and instrumental constant, thus providing a "self-calibrating" technique in the same manner relied upon by the occultation sensors on satellites. Moreover, the ground-based Umkehr measurement is the only technique that provides data with the same altitude resolution and in the same units (DU) as do the UV-nadir instruments (SBUV-2, GOME-2, OMPS-nadir), i.e., as ozone amount in pressure layers, whereas, occultation instruments measure ozone density with height. A new Umkehr algorithm will enhance the information content of the retrieved profiles and extend the applicability of the technique. Automated Dobson and Brewer instruments offer the potential for greatly expanded network of Umkehr observations once the new algorithm is applied. We will discuss the new algorithm development and present results of its performance in comparisons of retrievals between co-located Brewer and Dobson ozone profiles measured at Arosa station in Switzerland.

Organic micropollutant removal from wastewater effluent-impacted drinking water sources during bank filtration and artificial recharge.

PubMed

Maeng, Sung Kyu; Ameda, Emmanuel; Sharma, Saroj K; Grützmacher, Gesche; Amy, Gary L

2010-07-01

Natural treatment systems such as bank filtration (BF) and artificial recharge (via an infiltration basin) are a robust barrier for many organic micropollutants (OMPs) and may represent a low-cost alternative compared to advanced drinking water treatment systems. This study analyzes a comprehensive database of OMPs at BF and artificial recharge (AR) sites located near Lake Tegel in Berlin (Germany). The focus of the study was on the derivation of correlations between the removal efficiencies of OMPs and key factors influencing the performance of BF and AR. At the BF site, shallow monitoring wells located close to the Lake Tegel source exhibited oxic conditions followed by prolonged anoxic conditions in deep monitoring wells and a production well. At the AR site, oxic conditions prevailed from the recharge pond along monitoring wells to the production well. Long residence times of up to 4.5 months at the BF site reduced the temperature variation during soil passage between summer and winter. The temperature variations were greater at the AR site as a consequence of shorter residence times. Deep monitoring wells and the production well located at the BF site were under the influence of ambient groundwater and old bank filtrate (up to several years of age). Thus, it is important to account for mixing with native groundwater and other sources (e.g., old bank filtrate) when estimating the performance of BF with respect to removal of OMPs. Principal component analysis (PCA) was used to investigate correlations between OMP removals and hydrogeochemical conditions with spatial and temporal parameters (e.g., well distance, residence time and depth) from both sites. Principal component-1 (PC1) embodied redox conditions (oxidation-reduction potential and dissolved oxygen), and principal component-2 (PC2) embodied degradation potential (e.g., total organic carbon and dissolved organic carbon) with the calcium carbonate dissolution potential (Ca(2+) and HCO(3)(-)) for the BF site. These two PCs explained a total variance of 55% at the BF site. At the AR site, PCA revealed redox conditions (PC1) and degradation potential with temperature (PC2) as principal components, which explained a total variance of 56%. Copyright 2010 Elsevier Ltd. All rights reserved.

A Lactococcus lactis BFE920 feed vaccine expressing a fusion protein composed of the OmpA and FlgD antigens from Edwardsiella tarda was significantly better at protecting olive flounder (Paralichthys olivaceus) from edwardsiellosis than single antigen vaccines.

PubMed

Beck, Bo Ram; Lee, Soon Ho; Kim, Daniel; Park, Ji Hye; Lee, Hyun Kyung; Kwon, San-Sung; Lee, Kwan Hee; Lee, Jae Il; Song, Seong Kyu

2017-09-01

Edwardsiellosis is a major fish disease that causes a significant economic damage in the aquaculture industry. Here, we assessed vaccine efficacy after feeding oral vaccines to olive flounder (Paralichthys olivaceus), either L. lactis BFE920 expressing Edwardsiella tarda outer membrane protein A (OmpA), flagellar hook protein D (FlgD), or a fusion antigen of the two. Feed vaccination was done twice with a one-week interval. Fish were fed regular feed adsorbed with the vaccines. Feed vaccination was given over the course of one week to maximize the interaction between the feed vaccines and the fish intestine. Flounder fed the vaccine containing the fusion antigen had significantly elevated levels T cell genes (CD4-1, CD4-2, and CD8α), type 1 helper T cell (Th1) subset indicator genes (T-bet and IFN-γ), and antigen-specific antibodies compared to the groups fed the single antigen-expressing vaccines. Furthermore, the superiority of the fusion vaccine was also observed in survival rates when fish were challenged with E. tarda: OmpA-FlgD-expressing vaccine (82.5% survival); FlgD-vaccine (55.0%); OmpA-vaccine (50%); WT L. lactis BFE920 (37.5%); Ctrl (10%). In addition, vaccine-fed fish exhibited increased weight gain (∼20%) and a decreased feed conversion ratio (∼20%) during the four week vaccination period. Flounder fed the FlgD-expressing vaccine, either the single or the fusion form, had significantly increased expression of TLR5M, IL-1β, and IL-12p40, suggesting that the FlgD may be a ligand of olive flounder TLR5M receptor or closely related to the TLR5M pathway. In conclusion, the present study demonstrated that olive flounder fed L. lactis BFE920 expressing a fusion antigen composed of E. tarda OmpA and FlgD showed a strong protective effect against edwardsiellosis indicating this may be developed as an E. tarda feed vaccine. Copyright © 2017 Elsevier Ltd. All rights reserved.

Reframing Resilience: Pilot Evaluation of a Program to Promote Resilience in Marginalized Older Adults

ERIC Educational Resources Information Center

Fullen, Matthew C.; Gorby, Sean R.

2016-01-01

Resilience has been described as a paradigm for aging that is more inclusive than models that focus on physiological and functional abilities. We evaluated a novel program, Resilient Aging, designed to influence marginalized older adults' perceptions of their resilience, self-efficacy, and wellness. The multiweek group program incorporated an…

A general program to compute the multivariable stability margin for systems with parametric uncertainty

NASA Technical Reports Server (NTRS)

Sanchez Pena, Ricardo S.; Sideris, Athanasios

1988-01-01

A computer program implementing an algorithm for computing the multivariable stability margin to check the robust stability of feedback systems with real parametric uncertainty is proposed. The authors present in some detail important aspects of the program. An example is presented using lateral directional control system.

Margins within Margins?: Voices Speaking through a Study of the Provision of an Educational Program for the Children of One Australian Show Circuit.

ERIC Educational Resources Information Center

Rose, C. G.; And Others

This paper examines the tactics used by the Showmen's Guild of Australasia in successfully lobbying for the development of a distance education program for their children. The Guild is considered to be a "marginalized" group, meaning members have less access to wealth, power, and status. Since 1930, members of the Showmen's Guild and…

Isolation and characterization of a novel Rickettsia species (Rickettsia asembonensis sp. nov.) obtained from cat fleas (Ctenocephalides felis).

PubMed

Maina, Alice N; Luce-Fedrow, Alison; Omulo, Sylvia; Hang, Jun; Chan, Teik-Chye; Ade, Fredrick; Jima, Dereje D; Ogola, Eric; Ge, Hong; Breiman, Robert F; Njenga, Moses K; Richards, Allen L

2016-11-01

A novel rickettsial agent, 'Candidatus Rickettsia asembonensis' strain NMRCiiT, was isolated from cat fleas, Ctenocephalides felis, from Kenya. Genotypic characterization of the new isolate based on sequence analysis of five rickettsial genes, rrs, gltA, ompA, ompB and sca4, indicated that this isolate clustered with Rickettsia felis URRWXCal2. The degree of nucleotide similarity demonstrated that isolate NMRCiiT belongs within the genus Rickettsia and fulfils the criteria for classification as a representative of a novel species. The name Rickettsia asembonensis sp. nov. is proposed, with NMRCiiT (=DSM 100172T=CDC CRIRC RAS001T=ATCC VR-1827T) as the type strain.

Operations mission planner beyond the baseline

NASA Technical Reports Server (NTRS)

Biefeld, Eric; Cooper, Lynne

1991-01-01

The scheduling of Space Station Freedom must satisfy four major requirements. It must ensure efficient housekeeping operations, maximize the collection of science, respond to changes in tasking and available resources, and accommodate the above changes in a manner that minimizes disruption of the ongoing operations of the station. While meeting these requirements the scheduler must cope with the complexity, scope, and flexibility of SSF operations. This requires the scheduler to deal with an astronomical number of possible schedules. The Operations Mission Planner (OMP) is centered around minimally disruptive replanning and the use of heuristics limit search in scheduling. OMP has already shown several artificial intelligence based scheduling techniques such as Interleaved Iterative Refinement and Bottleneck Identification using Process Chronologies.

The NASA Applied Sciences Program: Volcanic Ash Observations and Applications

NASA Technical Reports Server (NTRS)

Murray, John J.; Fairlie, Duncan; Green, David; Haynes, John; Krotkov, Nickolai; Meyer, Franz; Pavolonis, Mike; Trepte, Charles; Vernier, Jean-Paul

2016-01-01

Since 2000, the NASA Applied Sciences Program has been actively transitioning observations and research to operations. Particular success has been achieved in developing applications for NASA Earth Observing Satellite (EOS) sensors, integrated observing systems, and operational models for volcanic ash detection, characterization, and transport. These include imager applications for sensors such as the MODerate resolution Imaging SpectroRadiometer (MODIS) on NASA Terra and Aqua satellites, and the Visible Infrared Imaging Radiometer Suite (VIIRS) on the NASA/NOAA Suomi NPP satellite; sounder applications for sensors such as the Atmospheric Infrared Sounder (AIRS) on Aqua, and the Cross-track Infrared Sounder (CrIS) on Suomi NPP; UV applications for the Ozone Mapping Instrument (OMI) on the NASA Aura Satellite and the Ozone Mapping Profiler Suite (OMPS) on Suomi NPP including Direct readout capabilities from OMI and OMPS in Alaska (GINA) and Finland (FMI):; and lidar applications from the Caliop instrument coupled with the imaging IR sensor on the NASA/CNES CALIPSO satellite. Many of these applications are in the process of being transferred to the Washington and Alaska Volcanic Ash Advisory Centers (VAAC) where they support operational monitoring and advisory services. Some have also been accepted, transitioned and adapted for direct, onboard, automated product production in future U.S. operational satellite systems including GOES-R, and in automated volcanic cloud detection, characterization and alerting tools at the VAACs. While other observations and applications remain to be developed for the current constellation of NASA EOS sensors and integrated with observing and forecast systems, future requirements and capabilities for volcanic ash observations and applications are also being developed. Many of these are based on technologies currently being tested on NASA aircraft, Unmanned Aerial Systems (UAS) and balloons. All of these efforts and the potential advances that will be realized by integrating them are shared in this presentation.

Automated design and optimization of flexible booster autopilots via linear programming. Volume 2: User's manual

NASA Technical Reports Server (NTRS)

Hauser, F. D.; Szollosi, G. D.; Lakin, W. S.

1972-01-01

COEBRA, the Computerized Optimization of Elastic Booster Autopilots, is an autopilot design program. The bulk of the design criteria is presented in the form of minimum allowed gain/phase stability margins. COEBRA has two optimization phases: (1) a phase to maximize stability margins; and (2) a phase to optimize structural bending moment load relief capability in the presence of minimum requirements on gain/phase stability margins.

Solvent-Free Self-Assembly to the Synthesis of Nitrogen-Doped Ordered Mesoporous Polymers for Highly Selective Capture and Conversion of CO2.

PubMed

Liu, Fujian; Huang, Kuan; Wu, Qin; Dai, Sheng

2017-07-01

A solvent-free induced self-assembly technology for the synthesis of nitrogen-doped ordered mesoporous polymers (N-OMPs) is developed, which is realized by mixing polymer precursors with block copolymer templates, curing at 140-180 °C, and calcination to remove the templates. This synthetic strategy represents a significant advancement in the preparation of functional porous polymers through a fast and scalable yet environmentally friendly route, since no solvents or catalysts are used. The synthesized N-OMPs and their derived catalysts are found to exhibit competitive CO 2 capacities (0.67-0.91 mmol g -1 at 25 °C and 0.15 bar), extraordinary CO 2 /N 2 selectivities (98-205 at 25 °C), and excellent activities for catalyzing conversion of CO 2 into cyclic carbonate (conversion >95% at 100 °C and 1.2 MPa for 1.5 h). The solvent-free technology developed in this work can also be extended to the synthesis of N-OMP/SiO 2 nanocomposites, mesoporous SiO 2 , crystalline mesoporous TiO 2 , and TiPO, demonstrating its wide applicability in porous material synthesis. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

From LIMS to OMPS-LP: limb ozone observations for future reanalyses

NASA Astrophysics Data System (ADS)

Wargan, K.; Kramarova, N. A.; Remsberg, E. E.; Coy, L.; Harvey, L.; Livesey, N. J.; Pawson, S.

2017-12-01

High vertical resolution and accuracy of ozone data from satellite-borne limb sounders have made them an invaluable tool in scientific studies of the middle and upper atmosphere. However, it was not until recently that these measurements were successfully incorporated in atmospheric reanalyses: of the major multidecadal reanalyses only ECMWF's ERA-Interim/ERA5 and NASA's MERRA-2 use limb ozone data. Validation and comparison studies have demonstrated that the addition of observations from the Microwave Limb Sounder (MLS) on EOS Aura greatly improved the quality of ozone fields in MERRA-2 making these assimilated data sets useful for scientific research. In this presentation, we will show the results of test experiments assimilating retrieved ozone from the Limb Infrared Monitor of the Stratosphere (LIMS, 1978/1979) and Ozone Mapping Profiler Suite Limb Profiler (OMPS-LP, 2012 to present). Our approach builds on the established assimilation methodology used for MLS in MERRA-2 and, in the case of OMPS-LP, extends the excellent record of MLS ozone assimilation into the post-EOS era in Earth observations. We will show case studies, discuss comparisons of the new experiments with MERRA-2, strategies for bias correction and the potential for combined assimilation of multiple limb ozone data types in future reanalyses for studies of multidecadal stratospheric ozone changes including trends.

Structure and inhibition of orotidine 5'-monophosphate decarboxylase from Plasmodium falciparum.

PubMed

Langley, David B; Shojaei, Maryam; Chan, Camilla; Lok, Hiu Chuen; Mackay, Joel P; Traut, Thomas W; Guss, J Mitchell; Christopherson, Richard I

2008-03-25

Orotidine 5'-monophosphate (OMP) decarboxylase from Plasmodium falciparum (PfODCase, EC 4.1.1.23) has been overexpressed, purified, subjected to kinetic and biochemical analysis, and crystallized. The native enzyme is a homodimer with a subunit molecular mass of 38 kDa. The saturation curve for OMP as a substrate conformed to Michaelis-Menten kinetics with K m = 350 +/- 60 nM and V max = 2.70 +/- 0.10 micromol/min/mg protein. Inhibition patterns for nucleoside 5'-monophosphate analogues were linear competitive with respect to OMP with a decreasing potency of inhibition of PfODCase in the order: pyrazofurin 5'-monophosphate ( K i = 3.6 +/- 0.7 nM) > xanthosine 5'-monophosphate (XMP, K i = 4.4 +/- 0.7 nM) > 6-azauridine 5'-monophosphate (AzaUMP, K i = 12 +/- 3 nM) > allopurinol-3-riboside 5'-monophosphate ( K i = 240 +/- 20 nM). XMP is an approximately 150-fold more potent inhibitor of PfODCase compared with the human enzyme. The structure of PfODCase was solved in the absence of ligand and displays a classic TIM-barrel fold characteristic of the enzyme. Both the phosphate-binding loop and the betaalpha5-loop have conformational flexibility, which may be associated with substrate capture and product release along the reaction pathway.

A targeted boost-and-sort immunization strategy using Escherichia coli BamA identifies rare growth inhibitory antibodies.

PubMed

Vij, Rajesh; Lin, Zhonghua; Chiang, Nancy; Vernes, Jean-Michel; Storek, Kelly M; Park, Summer; Chan, Joyce; Meng, Y Gloria; Comps-Agrar, Laetitia; Luan, Peng; Lee, Sophia; Schneider, Kellen; Bevers, Jack; Zilberleyb, Inna; Tam, Christine; Koth, Christopher M; Xu, Min; Gill, Avinash; Auerbach, Marcy R; Smith, Peter A; Rutherford, Steven T; Nakamura, Gerald; Seshasayee, Dhaya; Payandeh, Jian; Koerber, James T

2018-05-08

Outer membrane proteins (OMPs) in Gram-negative bacteria are essential for a number of cellular functions including nutrient transport and drug efflux. Escherichia coli BamA is an essential component of the OMP β-barrel assembly machinery and a potential novel antibacterial target that has been proposed to undergo large (~15 Å) conformational changes. Here, we explored methods to isolate anti-BamA monoclonal antibodies (mAbs) that might alter the function of this OMP and ultimately lead to bacterial growth inhibition. We first optimized traditional immunization approaches but failed to identify mAbs that altered cell growth after screening >3000 hybridomas. We then developed a "targeted boost-and-sort" strategy that combines bacterial cell immunizations, purified BamA protein boosts, and single hybridoma cell sorting using amphipol-reconstituted BamA antigen. This unique workflow improves the discovery efficiency of FACS + mAbs by >600-fold and enabled the identification of rare anti-BamA mAbs with bacterial growth inhibitory activity in the presence of a truncated lipopolysaccharide layer. These mAbs represent novel tools for dissecting the BamA-mediated mechanism of β-barrel folding and our workflow establishes a new template for the efficient discovery of novel mAbs against other highly dynamic membrane proteins.

Fed batch fermentation and purification strategy for high yield production of Brucella melitensis recombinant Omp 28 kDa protein and its application in disease diagnosis.

PubMed

Karothia, B S; Athmaram, T N; D, Thavaselvam; Ashu, Kumar; Tiwari, Sapna; Singh, Anil K; Sathyaseelan, K; Gopalan, N

2013-07-01

Brucellosis is a disease caused by bacteria belonging to the genus Brucella. It affects cattle, goat, sheep, dog and humans. The serodiagnosis of brucellosis involves detection of antibodies generated against the LPS or whole cell bacterial extracts, however these tests lack sensitivity and specificity. The present study was performed to optimize the culture condition for the production of recombinant Brucella melitensis outer membrane protein 28 kDa protein in E.coli via fed batch fermentation. Expression was induced with 1.5mM isopropyl β thiogalactoside and the expressed recombinant protein was purified using Ni-NTA affinity chromatography. After fed-batch fermentation the dry cell weight of 17.81 g/L and a purified protein yield of 210.10 mg/L was obtained. The purified Brucella melitensis recombinant Omp 28 kDa protein was analyzed through SDS- poly acrylamide gel electrophoresis and western blotting. The obtained recombinant protein was evaluated for its diagnostic application through Indirect ELISA using brucellosis suspected human sera samples. Our results clearly indicate that recombinant Omp28 produced via fed batch fermentation has immense potential as a diagnostic reagent that could be employed in sero monitoring of brucellosis.

Analysis of major antigens of Haemophilus (Actinobacillus) pleuropneumoniae and related organisms.

PubMed Central

MacInnes, J I; Rosendal, S

1987-01-01

Outer membrane protein (OMP)-enriched extracts and whole-cell protein preparations of Haemophilus (Actinobacillus) pleuropneumoniae and related organisms were examined by polyacrylamide gel electrophoresis and immunoblotting. Both the OMP-enriched and whole-cell protein profiles of Actinobacillus suis, A. pleuropneumoniae (NAD-independent biovar), A. lignieresii, and Pasteurella haemolytica were very similar to those of H. pleuropneumoniae serotypes 1 to 8. Antisera prepared against H. pleuropneumoniae typically recognized three major OMP antigens with approximate molecular weights of 17,000 (17K), 32K, and 42K in immunoblots of H. pleuropneumoniae serotypes 1 to 8, Actinobacillus spp., and P. haemolytica. Antisera prepared against Actinobacillus spp. and Haemophilus sp. "minor group" also recognized these 17K, 32K, and 42K antigens. Using absorbed sera, we demonstrated that the 17K antigen had an epitope (or epitopes) common to all the gram-negative organisms examined, including Escherichia coli. The 32K and 42K antigens had epitopes common to members of the family Pasteurellaceae but, in the case of the 32K antigen, also contained unique epitopes. These results provide a basis for understanding the lack of specificity of serodiagnostic tests for H. pleuropneumoniae infection and provide another line of evidence for the association of H. pleuropneumoniae with the genus Actinobacillus. Images PMID:3298061

Immunoproteomic Analysis To Identify Shiga Toxin-Producing Escherichia coli Outer Membrane Proteins Expressed during Human Infection

PubMed Central

Montero, David; Orellana, Paz; Gutiérrez, Daniela; Araya, Daniela; Salazar, Juan Carlos; Prado, Valeria; Oñate, Ángel; del Canto, Felipe

2014-01-01

Shiga-toxin producing Escherichia coli (STEC) is the etiologic agent of acute diarrhea, dysentery, and hemolytic-uremic syndrome (HUS). There is no approved vaccine for STEC infection in humans, and antibiotic use is contraindicated, as it promotes Shiga toxin production. In order to identify STEC-associated antigens and immunogenic proteins, outer membrane proteins (OMPs) were extracted from STEC O26:H11, O103, O113:H21, and O157:H7 strains, and commensal E. coli strain HS was used as a control. SDS-PAGE, two-dimensional-PAGE analysis, Western blot assays using sera from pediatric HUS patients and controls, and matrix-assisted laser desorption ionization–tandem time of flight analyses were used to identify 12 immunogenic OMPs, some of which were not reactive with control sera. Importantly, seven of these proteins have not been previously reported to be immunogenic in STEC strains. Among these seven proteins, OmpT and Cah displayed IgG and IgA reactivity with sera from HUS patients. Genes encoding these two proteins were present in a majority of STEC strains. Knowledge of the antigens produced during infection of the host and the immune response to those antigens will be important for future vaccine development. PMID:25156722

A lower size limit exists for export of fragments of an outer membrane protein (OmpA) of Escherichia coli K-12.

PubMed

Freudl, R; Schwarz, H; Degen, M; Henning, U

1989-02-20

The ompA gene codes for a 346 residue precursor of a 325 residue protein of the outer membrane of Escherichia coli K-12. Internally and/or COOH-terminally deleted genes were constructed that encode 123, 116, 88, 72 or 68 residue precursors. The former three were processed and localized to the periplasmic space; the latter two were not processed and remained cytosolic. These data suggest that the signal sequence has to interact with a component of the export apparatus (the Sec pathway) before translation is finished. Comparison of these results with others obtained for prokaryotic and eukaryotic systems shows that: (1) a very similar lower size limit exists for membrane translocation of the 147 residue chicken prelysozyme or the 229 residue bovine preprolactin; (2) precursors smaller than those reported here can be translocated in both systems; (3) the latter translocation, in contrast to, for example, the ompA gene products, does not depend on the cellular export machinery but most likely requires folding of the precursors into an export-competent conformation. In general, at least two quite different, not necessarily mutually exclusive, mechanisms for translocation of a protein across or assembly into a membrane appear to exist.

Proteomic analysis of Brucella abortus cell envelope and identification of immunogenic candidate proteins for vaccine development.

PubMed

Connolly, Joseph P; Comerci, Diego; Alefantis, Timothy G; Walz, Alexander; Quan, Marian; Chafin, Ryan; Grewal, Paul; Mujer, Cesar V; Ugalde, Rodolfo A; DelVecchio, Vito G

2006-07-01

Brucella abortus is the etiologic agent of bovine brucellosis and causes a chronic disease in humans known as undulant fever. In livestock the disease is characterized by abortion and sterility. Live, attenuated vaccines such as S19 and RB51 have been used to control the spread of the disease in animals; however, they are considered unsafe for human use and they induce abortion in pregnant cattle. For the development of a safer and equally efficacious vaccine, immunoproteomics was utilized to identify novel candidate proteins from B. abortus cell envelope (CE). A total of 163 proteins were identified using 2-DE with MALDI-TOF MS and LC-MS/MS. Some of the major protein components include outer-membrane protein (OMP) 25, OMP31, Omp2b porin, and 60 kDa chaperonin GroEL. 2-DE Western blot analyses probed with antiserum from bovine and a human patient infected with Brucella identified several new immunogenic proteins such as fumarate reductase flavoprotein subunit, F0F1-type ATP synthase alpha subunit, and cysteine synthase A. The elucidation of the immunome of B. abortus CE identified a number of candidate proteins for developing vaccines against Brucella infection in bovine and humans.

Vaccination with recombinant L7/L12-truncated Omp31 protein induces protection against Brucella infection in BALB/c mice.

PubMed

Golshani, Maryam; Rafati, Sima; Dashti, Amir; Gholami, Elham; Siadat, Seyed Davar; Oloomi, Mana; Jafari, Anis; Bouzari, Saeid

2015-06-01

Brucellosis is the most common bacterial zoonotic disease worldwide and no vaccine is available for the prevention of human brucellosis. In humans, brucellosis is mostly caused by Brucella melitensis and Brucella abortus. The Outer membrane protein 31 (Omp31) and L7/L12 are immunodominant and protective antigens conserved in human Brucella pathogens. In the present study, we evaluated the humoral and cellular immune responses induced by a fusion protein designed based on the Truncated form of Omp31 (TOmp31) and L7-L12 antigens. Vaccination of BALB/c mice with the recombinant fusion protein (rL7/L12-TOmp31) provided the significant protection level against B. melitensis and B. abortus challenge. Moreover, rL7/L12-TOmp31 elicited a strong specific IgG response (higher IgG2a titers) and significant IFN-γ/IL2 production and T-cell proliferation was also observed. The T helper1 (Th1) oriented response persisted for 12 weeks after the first immunization. The rL7/L12-TOmp31 could be a new potential antigen candidate for the development of a subunit vaccine against B. melitensis and B. abortus. Copyright © 2015 Elsevier Ltd. All rights reserved.

Aerosol layer height from synergistic use of VIIRS and OMPS

NASA Astrophysics Data System (ADS)

Lee, J.; Hsu, N. Y. C.; Sayer, A. M.; Kim, W.; Seftor, C. J.

2017-12-01

This study presents an Aerosol Single-scattering albedo and Height Estimation (ASHE) algorithm, which retrieves the height of UV-absorbing aerosols by synergistically using the Visible Infrared Imaging Radiometer Suite (VIIRS) and the Ozone Mapping and Profiler Suite (OMPS). ASHE provides height information over a much broader area than ground-based or spaceborne lidar measurements by benefitting from the wide swaths of the two instruments used. As determination of single-scattering albedo (SSA) of the aerosol layer is the most critical part for the performance and coverage of ASHE, here we demonstrate three different strategies to constrain the SSA. First, ASHE is able to retrieve the SSA of UV-absorbing aerosols when Cloud-Aerosol Lidar with Orthogonal Polarization (CALIOP) provides vertical profiles of the aerosol layer of interest. Second, Aerosol Robotic Network (AERONET) inversions can directly constrain the SSA of the aerosol layer when collocated with VIIRS or OMPS. Last, a SSA climatology from ASHE, AERONET, or other data sources can be used for large-scale, aged aerosol events, for which climatological SSA is well-known, at the cost of a slight decrease in retrieval accuracy. The same algorithm can be applied to measurements of similar type, such as those made by the Moderate Resolution Imaging Spectroradiometer (MODIS) and Ozone Monitoring Instrument (OMI), for a long-term, consistent data record.

Subinhibitory Concentrations of Antimicrobial Agents Reduce the Uptake of Legionella pneumophila into Acanthamoeba castellanii and U937 Cells by Altering the Expression of Virulence-Associated Antigens

PubMed Central

Lück, P. Christian; Schmitt, Jürgen W.; Hengerer, Arne; Helbig, Jürgen H.

1998-01-01

We determined the MICs of ampicillin, ciprofloxacin, erythromycin, imipenem, and rifampin for two clinical isolates of Legionella pneumophila serogroup 1 by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay and by quantitative culture. To test the influence of subinhibitory concentrations (sub-MICs) of antimicrobial agents on Legionella uptake into Acanthamoeba castellanii and U937 macrophage-like cells, both strains were pretreated with 0.25 MICs of the antibiotics for 24 h. In comparison to that for the untreated control, subinhibitory concentrations of antibiotics significantly reduced Legionella uptake into the host cells. Measurement of the binding of monoclonal antibodies against several Legionella antigens by enzyme-linked immunoassays indicated that sub-MIC antibiotic treatment reduced the expression of the macrophage infectivity potentiator protein (Mip), the Hsp 60 protein, the outer membrane protein (OmpM), an as-yet-uncharacterized protein of 55 kDa, and a few lipopolysaccharide (LPS) epitopes. In contrast, the expression of some LPS epitopes recognized by monoclonal antibodies 8/5 and 30/4 as well as a 45-kDa protein, a 58-kDa protein, and the major outer membrane protein (OmpS) remained unaffected. PMID:9797218

Ionic liquid biodegradability depends on specific wastewater microbial consortia.

PubMed

Docherty, Kathryn M; Aiello, Steven W; Buehler, Barbara K; Jones, Stuart E; Szymczyna, Blair R; Walker, Katherine A

2015-10-01

Complete biodegradation of a newly-synthesized chemical in a wastewater treatment plant (WWTP) eliminates the potential for novel environmental pollutants. However, differences within- and between-WWTP microbial communities may alter expectations for biodegradation. WWTP communities can also serve as a source of unique consortia that, when enriched, can metabolize chemicals that tend to resist degradation, but are otherwise promising green alternatives. We tested the biodegradability of three ionic liquids (ILs): 1-octyl-3-methylpyridinium bromide (OMP), 1-butyl-3-methylpyridinium bromide (BMP) and 1-butyl-3-methylimidazolium chloride (BMIM). We performed tests using communities from two WWTPs at three time points. Site-specific and temporal variation both influenced community composition, which impacted the success of OMP biodegradability. Neither BMP nor BMIM degraded in any test, suggesting that these ILs are unlikely to be removed by traditional treatment. Following standard biodegradation assays, we enriched for three consortia that were capable of quickly degrading OMP, BMP and BMIM. Our results indicate WWTPs are not functionally redundant with regard to biodegradation of specific ionic liquids. However, consortia can be enriched to degrade chemicals that fail biodegradability assays. This information can be used to prepare pre-treatment procedures and prevent environmental release of novel pollutants. Copyright © 2015 Elsevier Ltd. All rights reserved.

A conserved OmpA-like protein in Legionella pneumophila required for efficient intracellular replication.

PubMed

Goodwin, Ian P; Kumova, Ogan K; Ninio, Shira

2016-08-01

The OmpA-like protein domain has been associated with peptidoglycan-binding proteins, and is often found in virulence factors of bacterial pathogens. The intracellular pathogen Legionella pneumophila encodes for six proteins that contain the OmpA-like domain, among them the highly conserved uncharacterized protein we named CmpA. Here we set out to characterize the CmpA protein and determine its contribution to intracellular survival of L. pneumophila Secondary structure analysis suggests that CmpA is an inner membrane protein with a peptidoglycan-binding domain at the C-teminus. A cmpA mutant was able to replicate normally in broth, but failed to compete with an isogenic wild-type strain in an intracellular growth competition assay. The cmpA mutant also displayed significant intracellular growth defects in both the protozoan host Acanthamoeba castellanii and in primary bone marrow-derived macrophages, where uptake into the cells was also impaired. The cmpA phenotypes were completely restored upon expression of CmpA in trans The data presented here establish CmpA as a novel virulence factor of L. pneumophila that is required for efficient intracellular replication in both mammalian and protozoan hosts. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Evolution of the Translocation and Assembly Module (TAM)

PubMed Central

Heinz, Eva; Selkrig, Joel; Belousoff, Matthew J.; Lithgow, Trevor

2015-01-01

Bacterial outer membrane proteins require the beta-barrel assembly machinery (BAM) for their correct folding and function. The central component of this machinery is BamA, an Omp85 protein that is essential and found in all Gram-negative bacteria. An additional feature of the BAM is the translocation and assembly module (TAM), comprised TamA (an Omp85 family protein) and TamB. We report that TamA and a closely related protein TamL are confined almost exclusively to Proteobacteria and Bacteroidetes/Chlorobi respectively, whereas TamB is widely distributed across the majority of Gram-negative bacterial lineages. A comprehensive phylogenetic and secondary structure analysis of the TamB protein family revealed that TamB was present very early in the evolution of bacteria. Several sequence characteristics were discovered to define the TamB protein family: A signal-anchor linkage to the inner membrane, beta-helical structure, conserved domain architecture and a C-terminal region that mimics outer membrane protein beta-strands. Taken together, the structural and phylogenetic analyses suggest that the TAM likely evolved from an original combination of BamA and TamB, with a later gene duplication event of BamA, giving rise to an additional Omp85 sequence that evolved to be TamA in Proteobacteria and TamL in Bacteroidetes/Chlorobi. PMID:25994932

Olfactory marker protein is critical for functional maturation of olfactory sensory neurons and development of mother preference

PubMed Central

Lee, Anderson C.; He, Jiwei; Ma, Minghong

2011-01-01

Survival of many altricial animals critically depends on the sense of smell. Curiously, the olfactory system is rather immature at birth and undergoes a maturation process, which is poorly understood. Using patch clamp technique on mouse olfactory sensory neurons (OSNs) with a defined odorant receptor (OR), we demonstrate that OSNs exhibit functional maturation during the first month of postnatal life by developing faster response kinetics, higher sensitivity, and most intriguingly, higher selectivity. OSNs expressing the receptor MOR23 are relatively broadly tuned in neonates and become selective detectors for the cognate odorant within two weeks. Remarkably, these changes are prevented by genetic ablation of olfactory marker protein (OMP), which is exclusively expressed in mature OSNs. Biochemical and pharmacological evidence supports that alteration in odorant-induced phosphorylation of signaling proteins underlie some of the OMP−/− phenotypes. Furthermore, in a novel behavioral assay in which the mouse pups are given a choice between the biological mother and another unfamiliar lactating female, wild-type pups prefer the biological mother, while OMP knockout pups fail to show preference. These results reveal that OSNs undergo an OMP-dependant functional maturation process that coincides with early development of the smell function, which is essential for pups to form preference for their mother. PMID:21414919

Rickettsia vini n. sp. (Rickettsiaceae) infecting the tick Ixodes arboricola (Acari: Ixodidae).

PubMed

Novakova, Marketa; Costa, Francisco B; Krause, Frantisek; Literak, Ivan; Labruna, Marcelo B

2016-08-26

Recently, a new rickettsia named 'Candidatus Rickettsia vini' belonging to the spotted fever group has been molecularly detected in Ixodes arboricola ticks in Spain, the Czech Republic, Slovakia and Turkey, with prevalence reaching up to 100 %. The aim of this study was to isolate this rickettsia in pure culture, and to describe it as a new Rickettsia species. A total of 148 ornitophilic nidicolous ticks Ixodes arboricola were collected in a forest near Breclav (Czech Republic) and examined for rickettsiae. Shell vial technique was applied to isolate rickettsiae in Vero cells. Rickettsial isolation was confirmed by optical microscopy and sequencing of partial sequences of the rickettsial genes gltA, ompA, ompB, and htrA. Laboratory guinea pigs and chickens were used for experimental infestations and infections. Animal blood sera were tested by immunofluorescence assay employing crude antigens of various rickettsiae. Rickettsia vini n. sp. was successfully isolated from three males of I. arboricola. Phylogenetic analysis of fragments of 1092, 590, 800, and 497 nucleotides of the gltA, ompA, ompB, and htrA genes, respectively, showed closest proximity of R. vini n. sp. to Rickettsia japonica and Rickettsia heilongjiangensis belonging to the spotted fever group. Experimental infection of guinea pigs and chickens with R. vini led to various levels of cross-reactions of R. vini-homologous antibodies with Rickettsia rickettsii, Rickettsia parkeri, 'Candidatus Rickettsia amblyommii', Rickettsia rhipicephali, Rickettsia bellii, and Rickettsia felis. Laboratory infestations by R. vini-infected I. arboricola larvae on chickens led to no seroconversion to R. vini n. sp., nor cross-reactions with R. rickettsii, R. parkeri, 'Ca. R. amblyommii', R. rhipicephali, R. bellii or R. felis. Our results suggest that R. vini n. sp. is possibly a tick endosymbiont, not pathogenic for guinea pigs and chickens. Regarding specific phenotypic characters and significant differences of DNA sequences in comparison to the most closely related species (R. japonica and R. heilongjiangensis), we propose to classify the isolate as a new species, Rickettsia vini.

New Rickettsia species in soft ticks Ornithodoros hasei collected from bats in French Guiana.

PubMed

Tahir, Djamel; Socolovschi, Cristina; Marié, Jean-Lou; Ganay, Gautier; Berenger, Jean-Michel; Bompar, Jean-Michel; Blanchet, Denis; Cheuret, Marie; Mediannikov, Oleg; Raoult, Didier; Davoust, Bernard; Parola, Philippe

2016-10-01

In French Guiana, located on the northeastern coast of South America, bats of different species are very numerous. The infection of bats and their ticks with zoonotic bacteria, especially Rickettsia species, is so far unknown. In order to improve knowledge of these zoonotic pathogens in this French overseas department, the presence and diversity of tick-borne bacteria was investigated with molecular tools in bat ticks. In the beginning of 2013, 32 bats were caught in Saint-Jean-du-Maroni, an area close to the coast of French Guiana, and the ticks of these animals were collected. A total of 354 larvae of Argasidae soft ticks (Ornithodoros hasei) from 12 bats (Noctilio albiventris) were collected and 107 of them were analysed. DNA was extracted from the samples and quantitative real-time PCR was carried out to detect Rickettsia spp., Bartonella spp., Borrelia spp. and Coxiella burnetii. All tested samples were negative for Bartonella spp., Borrelia spp. and Coxiella burnetii. Rickettsia DNA was detected in 31 (28.9%) ticks. An almost entire (1118 base pairs long) sequence of the gltA gene was obtained after the amplification of some positive samples on conventional PCR and sequencing. A Bayesian tree was constructed using concatenated rrs, gltA, ompA, ompB, and gene D sequences. The study of characteristic sequences shows that this Rickettsia species is very close (98.3-99.8%) genetically to R. peacockii. Nevertheless, the comparative analysis of sequences obtained from gltA, ompA, ompB, rrs and gene D fragments demonstrated that this Rickettsia is different from the other members of the spotted fever group. The sequences of this new species were deposited in GenBank as Candidatus Rickettsia wissemanii. This is the first report showing the presence of nucleic acid of Rickettsia in Ornithodoros hasei ticks from South American bats. Copyright © 2016 Elsevier GmbH. All rights reserved.

Potential Applications of JNPP to Infrared-Based Remote Sensing of Volcanic Emissions

NASA Astrophysics Data System (ADS)

Realmuto, V. J.

2016-12-01

The simultaneous collection of VIIRS, CrIS, and OMPS data will make JNPP an ideal platform for monitoring volcanic emissions. For daytime overpasses we will obtain three contemporaneous, but independent, estimates of SO2 column density, as well as information on the quantity and composition of aerosols and volcanic ash. We will use the independent measurements to validate individual retrieval techniques, and exploit the synergy between UV and TIR remote sensing. The finer spatial resolution of VIIRS (750 m at nadir), relative to OMPS (50 km) and CrIS (14 km), will allow us to characterize variations in surface conditions, plume composition, and the distribution of clouds within an IFOV of CrIS or OMPS, and assess the impact of these variations on the SO2retrievals. Atmospheric profiles are an essential input to the retrieval procedures, and the profiles derived from CrIS soundings will provide us with an accurate description of atmospheric conditions local to the plumes. In addition, the fine spectral resolution of CrIS will enable us to identify and quantify the components of heterogeneous (gas + particulate) plumes. We will demonstrate the potential use of JPSS to map volcanic planes through the analyses of TIR data acquired by EOS (ASTER, MODIS, and AIRS) and SNPP (VIIRS and CrIS) instruments over the plumes generated by recent eruptions of Eyjafallajökull, Bardarbunga (Iceland), Calbuco (Chile), and Ontake (Japan) Volcanoes. We will present comparisons of the TIR-based retrievals to OMI and SNPP-OMPS data products. Finally, we will outline a path to operations through collaboration with the Alaska Volcano Observatory (USGS), Anchorage Volcanic Ash Advisory Center (NWS + FAA), NASA-GSFC Direct Readout Lab, and University of Alaska-Fairbanks. This research was conducted at the Jet Propulsion Laboratory, California Institute of Technology, under contract to National Atmospheric and Space Administration.

Satellite Observations of Tropospheric BrO over Salt Lakes and Northern High Latitudes from EOS/OMI and SNPP/OMPS

NASA Astrophysics Data System (ADS)

Kurosu, T. P.; Stutz, J.; Brockway, N.; Saiz-Lopez, A.; Suleiman, R. M.; Natraj, V.; Jaross, G.; Seftor, C. J.

2017-12-01

We present observations of tropospheric bromine monoxide (BrO) derived from two satellite instruments: the Ozone Monitoring Instrument (OMI) on EOS-Aura, and the Nadir Mapper component of the Ozone Mapping and Profiler Suite (OMPS) on Suomi/NPP. BrO observations from OMPS constitute a new and experimental measurement that we first report on here and compare with the standard BrO data product from OMI. BrO is a halogen oxide present mostly in the lower stratosphere, where it catalytically destroys ozone with about 25 times the efficiency of ClO. BrO also has a tropospheric component, where it is released from sea surfaces, at the interface of ocean water and sea ice in the polar spring, in volcanic plumes, and in the vicinity of salt lakes. Tropospheric BrO has been linked to mercury (Hg) deposition through BrO-induced conversion of gaseous Hg to reactive Hg, which is then deposited on the surface and enters the food chain, ultimately affecting human health. As part of NASA's Aura Science Team, we are developing an OMI Tropospheric BrO data product that provides a unique global data set on BrO spatial and vertical distribution in the troposphere and stratosphere. Information of this kind is currently unavailable from any of the past and present bromine-monitoring instruments. In this presentation, we focus on multi-year time series of BrO released from a range of salt lakes - the Rann of Kutch, Salar de Uyuni, the Aral Sea, and others. We quantify the amount of bromine released from the lakes and investigate the possibility of lake desiccation monitoring based on independent BrO observations. The quality and limits of OMI and OMPS tropospheric BrO observations is investigated by comparison with ground-based MAX-DOAS observations over central Greenland.

Study on the resistance mechanism via outer membrane protein OprD2 and metal β-lactamase expression in the cell wall of Pseudomonas aeruginosa.

PubMed

Cai, Shuangqi; Chen, Yiqiang; Song, Dezhi; Kong, Jinliang; Wu, Yanbin; Lu, Huasong

2016-11-01

The aim of the present study was to evaluate the imipenem-resistant mechanism via the outer membrane protein (OMP) OprD2 and metal β-lactamase expression in the cell wall of Pseudomonas aeruginosa . The Pseudomonas aeruginosa was clinically separated and validated by VITEK-2 full-automatic bacteria analyzer. Drug resistance, sensitive antibiotics and minimum inhibitory concentration (MIC) were tested using the drug sensitivity analysis system. The phenotype positive strains of MBL genes were screened using the Kirby-Bauer diffusion method by adding metal ion-chelating agent EDTA on the imipenem susceptibility paper. IMP-1, VIM-1 and SPM metaloenzyme genes were tested by polymerase chain reaction (PCR)-telomeric repeat amplification protocol (TRAP). The OMP OprD2 genes were tested by PCR-TRAP, and the protein expression was tested using western blot analysis. The location of OMP OprD2 was confirmed using the sodium salicylate inhibition test. The results showed that 80 portions (40%) of MBL-positive strains were screened out of 200 specimens. Imipenem-resistant Pseudomonas aeruginosa (IRPA) and MIC values were significantly higher than quality control bacteria and control bacteria (P<0.05). A total of 35 cases with IMP-1 positive, 20 with VIM-1 positive, 16 with SPM positive, 5 with 2 positive genes and 4 with 3 positive genes were screened among MBL positive strains. A total of 150 portions (75%) of OprD2 deficiencies were screened from 200 specimens. The standard strains and sensitive strains showed OprD2 protein bands at 45 kDa while no OprD2 protein bands appeared in OprD2 deficiency strains. It was in accordance with gene detection. In conclusion, OMP OprD2 deficiency and MBL phenotype positivity may be important mechanisms of IRPA.

Characterization of the Leptospiral Outer Membrane and Description of Three Novel Leptospiral Membrane Proteins

PubMed Central

Haake, David A.; Matsunaga, James

2002-01-01

The outer membrane (OM) of the mammalian pathogen Leptospira kirschneri was isolated in the form of membrane vesicles by alkaline plasmolysis and separated from the protoplasmic cylinder by sucrose density gradient ultracentrifugation. All four components of the alkaline plasmolysis buffer, including 1.0 M NaCl, 27% sucrose (wt/vol), 2 mM EDTA, and 10 mM Tris (pH 9), were required for efficient OM release, as judged by recovery of leptospiral lipopolysaccharide. Two populations of OM vesicles (OMVs) were recovered, with peak concentrations found in the sucrose gradient at densities of 1.16 and 1.18 g/ml. Transmission electron microscopy revealed that the more buoyant OMV population was smaller (<0.1 μm in diameter) than the denser OMV population (0.2 to 0.3 μm in diameter). The densities of both populations of OMVs were distinct from that of the protoplasmic-cylinder material, which was found in the sucrose gradient at a density of 1.20 g/ml. The OMV fractions were free of protoplasmic-cylinder material, as judged by immunoblotting with antibodies to the endoflagellar sheath protein, heat shock protein GroEL, and two novel cytoplasmic membrane proteins, lipoprotein LipL31 and transmembrane protein ImpL63. The protein components of the OMVs were characterized by one- and two-dimensional immunoblotting and found to include previously described OM proteins (OMPs), including the porin OmpL1; the lipoproteins LipL32, LipL36, and LipL41; and the peripheral membrane protein P31LipL45. A number of less well-characterized OMPs were also identified, including those with molecular masses of 16, 21, 21.5, 22, 31, 36, 44, 48, 90, and 116 kDa. The 48-kDa OMP was identified as a novel OM lipoprotein designated LipL48. The use of membrane-specific markers in OM isolation techniques facilitates an accurate description of the leptospiral OM and its components. PMID:12183539

The interaction of N-glycans in Fcγ receptor I α-chain with Escherichia coli K1 outer membrane protein A for entry into macrophages: experimental and computational analysis.

PubMed

Krishnan, Subramanian; Liu, Fan; Abrol, Ravinder; Hodges, Jacqueline; Goddard, William A; Prasadarao, Nemani V

2014-11-07

Neonatal meningitis, caused by Escherichia coli K1, is a serious central nervous system disease. We have established that macrophages serve as permissive niches for E. coli K1 to multiply in the host and for attaining a threshold level of bacterial load, which is a prerequisite for the onset of the disease. Here, we demonstrate experimentally that three N-glycans in FcγRIa interact with OmpA of E. coli K1 for binding to and entering the macrophages. Adoptive transfer of FcγRIa(-/-) bone marrow-derived macrophages transfected with FcγRIa into FcγRIa(-/-) newborn mice renders them susceptible to E. coli K1-induced meningitis. In contrast, mice that received bone marrow-derived macrophages transfected with FcγRIa in which N-glycosylation sites 1, 4, and 5 are mutated to alanines exhibit resistance to E. coli K1 infection. Our molecular dynamics and simulation studies predict that N-glycan 5 exhibits strong binding at the barrel site of OmpA formed by loops 3 and 4, whereas N-glycans 1 and 4 interact with loops 1, 3, and 4 of OmpA at tip regions. Molecular modeling data also suggest no role for the IgG binding site in the invasion process. In agreement, experimental mutations in IgG binding site had no effect on the E. coli K1 entry into macrophages in vitro or on the onset of meningitis in newborn mice. Together, this integration of experimental and computational studies reveals how the N-glycans in FcγRIa interact with the OmpA of E. coli K1 for inducing the disease pathogenesis. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

Organization of the main olfactory bulbs of some mammals: musk shrews, moles, hedgehogs, tree shrews, bats, mice, and rats.

PubMed

Kosaka, Katsuko; Kosaka, Toshio

2004-04-19

We immunohistochemically examined the organization of the main olfactory bulbs (MOBs) in seven mammalian species, including moles, hedgehogs, tree shrews, bats, and mice as well as laboratory musk shrews and rats. We focused our investigation on two points: 1) whether nidi, particular spheroidal synaptic regions subjacent to glomeruli, which we previously reported for the laboratory musk shrew MOBs, are also present in other animals and 2) whether the compartmental organization of glomeruli and two types of periglomerular cells we proposed for the rat MOBs are general in other animals. The general laminar pattern was similar among these seven species, but discrete nidi and the nidal layer were recognized only in two insectivores, namely, the mole and laboratory musk shrew. Olfactory marker protein-immunoreactive (OMP-IR) axons extended beyond the limits of the glomerular layer (GL) into the superficial region of the external plexiform layer (EPL) or the nidal layer in the laboratory musk shrew, mole, hedgehog, and tree shrew but not in bat, mouse, and rat. We observed, in nidi and the nidal layer in the mole and laboratory musk shrew MOBs, only a few OMP-IR axons. In the hedgehog, another insectivore, OMP-IR processes extending from the glomeruli were scattered and intermingled with calbindin D28k-IR cells at the border between the GL and the EPL. In the superficial region of the EPL of the tree shrew MOBs, there were a small number of tiny glomerulus-like spheroidal structures where OMP-IR axons protruding from glomeruli were intermingled with dendritic branches of surrounding calbindin D28k-IR cells. Furthermore, we recognized the compartmental organization of glomeruli and two types of periglomerular cells in the MOBs of all of the mammals we examined. These structural features are therefore considered to be common and important organizational principles of the MOBs. Copyright 2004 Wiley-Liss, Inc.

Rate and Equilibrium Constants for an Enzyme Conformational Change during Catalysis by Orotidine 5'-Monophosphate Decarboxylase.

PubMed

Goryanova, Bogdana; Goldman, Lawrence M; Ming, Shonoi; Amyes, Tina L; Gerlt, John A; Richard, John P

2015-07-28

The caged complex between orotidine 5'-monophosphate decarboxylase (ScOMPDC) and 5-fluoroorotidine 5'-monophosphate (FOMP) undergoes decarboxylation ∼300 times faster than the caged complex between ScOMPDC and the physiological substrate, orotidine 5'-monophosphate (OMP). Consequently, the enzyme conformational changes required to lock FOMP at a protein cage and release product 5-fluorouridine 5'-monophosphate (FUMP) are kinetically significant steps. The caged form of ScOMPDC is stabilized by interactions between the side chains from Gln215, Tyr217, and Arg235 and the substrate phosphodianion. The control of these interactions over the barrier to the binding of FOMP and the release of FUMP was probed by determining the effect of all combinations of single, double, and triple Q215A, Y217F, and R235A mutations on kcat/Km and kcat for turnover of FOMP by wild-type ScOMPDC; its values are limited by the rates of substrate binding and product release, respectively. The Q215A and Y217F mutations each result in an increase in kcat and a decrease in kcat/Km, due to a weakening of the protein-phosphodianion interactions that favor fast product release and slow substrate binding. The Q215A/R235A mutation causes a large decrease in the kinetic parameters for ScOMPDC-catalyzed decarboxylation of OMP, which are limited by the rate of the decarboxylation step, but much smaller decreases in the kinetic parameters for ScOMPDC-catalyzed decarboxylation of FOMP, which are limited by the rate of enzyme conformational changes. By contrast, the Y217A mutation results in large decreases in kcat/Km for ScOMPDC-catalyzed decarboxylation of both OMP and FOMP, because of the comparable effects of this mutation on rate-determining decarboxylation of enzyme-bound OMP and on the rate-determining enzyme conformational change for decarboxylation of FOMP. We propose that kcat = 8.2 s(-1) for decarboxylation of FOMP by the Y217A mutant is equal to the rate constant for cage formation from the complex between FOMP and the open enzyme, that the tyrosyl phenol group stabilizes the closed form of ScOMPDC by hydrogen bonding to the substrate phosphodianion, and that the phenyl group of Y217 and F217 facilitates formation of the transition state for the rate-limiting conformational change. An analysis of kinetic data for mutant enzyme-catalyzed decarboxylation of OMP and FOMP provides estimates for the rate and equilibrium constants for the conformational change that traps FOMP at the enzyme active site.

Purification and Bicelle Crystallization for Structure Determination of the E. coli Outer Membrane Protein TamA.

PubMed

Gruss, Fabian; Hiller, Sebastian; Maier, Timm

2015-01-01

TamA is an Omp85 protein involved in autotransporter assembly in the outer membrane of Escherichia coli. It comprises a C-terminal 16-stranded transmembrane β-barrel as well as three periplasmic POTRA domains, and is a challenging target for structure determination. Here, we present a method for crystal structure determination of TamA, including recombinant expression in E. coli, detergent extraction, chromatographic purification, and bicelle crystallization in combination with seeding. As a result, crystals in space group P21212 are obtained, which diffract to 2.3 Å resolution. This protocol also serves as a template for structure determination of other outer membrane proteins, in particular of the Omp85 family.

Differential in vivo gene expression of major Leptospira proteins in resistant or susceptible animal models.

PubMed

Matsui, Mariko; Soupé, Marie-Estelle; Becam, Jérôme; Goarant, Cyrille

2012-09-01

Transcripts of Leptospira 16S rRNA, FlaB, LigB, LipL21, LipL32, LipL36, LipL41, and OmpL37 were quantified in the blood of susceptible (hamsters) and resistant (mice) animal models of leptospirosis. We first validated adequate reference genes and then evaluated expression patterns in vivo compared to in vitro cultures. LipL32 expression was downregulated in vivo and differentially regulated in resistant and susceptible animals. FlaB expression was also repressed in mice but not in hamsters. In contrast, LigB and OmpL37 were upregulated in vivo. Thus, we demonstrated that a virulent strain of Leptospira differentially adapts its gene expression in the blood of infected animals.

Draft Genome Sequence of a Virulent Strain of Pasteurella Multocida Isolated From Alpaca

PubMed Central

Hurtado, Raquel Enma; Aburjaile, Flavia; Mariano, Diego; Canário, Marcus Vinicius; Benevides, Leandro; Fernandez, Daniel Antonio; Allasi, Nataly Olivia; Rimac, Rocio; Juscamayta, Julio Eduardo; Maximiliano, Jorge Enrique; Rosadio, Raul Hector; Azevedo, Vasco; Maturrano, Lenin

2017-01-01

Pasteurella multocida is one of the most frequently isolated bacteria in acute pneumonia cases, being responsible for high mortality rates in Peruvian young alpacas, with consequent social and economic costs. Here we report the genome sequence of P. multocida strain UNMSM, isolated from the lung of an alpaca diagnosed with pneumonia, in Peru. The genome consists of 2,439,814 base pairs assembled into 82 contigs and 2,252 protein encoding genes, revealing the presence of known virulence-associated genes (ompH, ompA, tonB, tbpA, nanA, nanB, nanH, sodA, sodC, plpB and toxA). Further analysis could provide insights about bacterial pathogenesis and control strategies of this disease in Peruvian alpacas. PMID:28698737

Adults in Programs for the "Academically Underprepared"

ERIC Educational Resources Information Center

Isserlis, Janet

2008-01-01

Adult basic education represents a marginalized field that addresses the needs of a largely marginalized population: adults who are viewed through a deficit lens labeling them as being underprepared. This article examines particular challenges and strengths of adult learners in basic education programs (including literacy, English language, and…

Brazilian spotted fever: real-time PCR for diagnosis of fatal cases.

PubMed

dos Santos, Fabiana Cristina Pereira; do Nascimento, Elvira Maria Mendes; Katz, Gizelda; Angerami, Rodrigo Nogueira; Colombo, Silvia; de Souza, Eliana Rodrigues; Labruna, Marcelo Bahia; da Silva, Marcos Vinicius

2012-12-01

Suspicion of Brazilian spotted fever (BSF) should occur in endemic regions upon surveillance of the acute febrile icteric hemorrhagic syndrome (AFIHS). However, limitations associated with currently available laboratory tests pose a challenge to early diagnosis, especially in fatal cases. Two real-time PCR (qPCR) protocols were evaluated to diagnose BSF in 110 fatal AFIHS cases, collected in BSF-endemic regions in 2009-2010. Of these, 24 were positive and 86 negative by indirect immunofluorescence (IFA) assay (cut-off IgG and/or IgM ≥ 128). DNA from these samples was used in the qPCR protocols: one to detect Rickettsia spp. (citrate synthase gene) and another to determine spotted fever group (SFG) Rickettsia species (OmpA gene). Of the 24 IFA-positive samples, 5 (21%) were positive for OmpA and 9 (38%) for citrate synthase. In the IFA-negative group (n=86), OmpA and citrate synthase were positive in 23 (27%) and 27 (31%), respectively. These results showed that the 2 qPCR protocols were about twice as sensitive as the IFA test alone (93% concordance). In conclusion, qPCR is a sensitive method for the diagnosis of fatal BSF cases and should be considered for routine surveillance of AFIHS in places like Brazil, where spotted fever-related lethality is high and other endemic diseases like dengue and leptospirosis can mislead diagnosis. Copyright © 2012 Elsevier GmbH. All rights reserved.

Evaluation of cell binding activities of Leptospira ECM adhesins.

PubMed

Robbins, Gregory T; Hahn, Beth L; Evangelista, Karen V; Padmore, Lavinia; Aranda, Patrick S; Coburn, Jenifer

2015-04-01

Pathogenic spirochetes of the genus Leptospira are the causative agents of leptospirosis, a zoonotic infection that occurs globally. The bacteria colonize the renal proximal tubules of many animals and are shed in the urine. Contact with the urine, or with water contaminated with the urine of infected animals can cause infection of new host animals, including humans. Mechanisms of colonization of the proximal tubule and other tissues are not known, but specific interactions between bacterial adhesins and host substrates are likely to be critical in this process. Several extracellular matrix (ECM) adhesins have been previously identified, but more recently, it has been shown that Leptospira bind more efficiently to cells than ECM. In this work, recombinant forms of five putative Leptospira ECM adhesins, namely LipL32, Loa22, OmpL1, p31/LipL45, and LenA were evaluated for binding to cells as well as an expanded variety of ECM components. Reproducible and significant adhesin activity was demonstrated only for OmpL1, which bound to both mammalian cell lines tested and to glycosaminoglycans (GAGs). While determination of biologically significant bacterial adhesion activity will require generation of site-directed mutant strains, our results suggest that OmpL1 is a strong candidate for future evaluation regarding the roles of the adhesin activity of the protein during L. interrogans infection.

Evaluation of Cell Binding Activities of Leptospira ECM Adhesins

PubMed Central

Robbins, Gregory T.; Hahn, Beth L.; Evangelista, Karen V.; Padmore, Lavinia; Aranda, Patrick S.; Coburn, Jenifer

2015-01-01

Pathogenic spirochetes of the genus Leptospira are the causative agents of leptospirosis, a zoonotic infection that occurs globally. The bacteria colonize the renal proximal tubules of many animals and are shed in the urine. Contact with the urine, or with water contaminated with the urine of infected animals can cause infection of new host animals, including humans. Mechanisms of colonization of the proximal tubule and other tissues are not known, but specific interactions between bacterial adhesins and host substrates are likely to be critical in this process. Several extracellular matrix (ECM) adhesins have been previously identified, but more recently, it has been shown that Leptospira bind more efficiently to cells than ECM. In this work, recombinant forms of five putative Leptospira ECM adhesins, namely LipL32, Loa22, OmpL1, p31/LipL45, and LenA were evaluated for binding to cells as well as an expanded variety of ECM components. Reproducible and significant adhesin activity was demonstrated only for OmpL1, which bound to both mammalian cell lines tested and to glycosaminoglycans (GAGs). While determination of biologically significant bacterial adhesion activity will require generation of site-directed mutant strains, our results suggest that OmpL1 is a strong candidate for future evaluation regarding the roles of the adhesin activity of the protein during L. interrogans infection. PMID:25875373

Detection of Iss and Bor on the surface of Escherichia coli.

PubMed

Lynne, A M; Skyberg, J A; Logue, C M; Nolan, L K

2007-03-01

To confirm the presence of Iss and Bor on the outer membrane of Escherichia coli using Western blots of outer membrane protein (OMP) preparations and fluorescence microscopy, and explore the use of fluorescence microscopy for the detection of avian pathogenic E. coli (APEC) and diagnosis of avian colibacillosis. Knockout mutants of iss and bor were created using a one-step recombination of target genes with PCR-generated antibiotic resistance cassettes. Anti-Iss monoclonal antibodies (Mabs) that cross-react with Bor protein were used to study the mutants relative to the wild-type organism. These Mabs were used as reagents to study OMP preparations of the mutants with Western blotting and intact E. coli cells with fluorescence microscopy. Iss and Bor were detected in Western blots of OMP preparations of the wild type. Also, Iss was detected on Deltabor mutants, and Bor was detected on Deltaiss mutants. Iss and Bor were also detected on the surface of the intact, wild-type cells and mutants using fluorescence microscopy. These results demonstrate that Bor and Iss are exposed on E. coli's outer membrane where they may be recognized by the host's immune system. To our knowledge, this is the first report confirming Iss' location in the outer membrane of an E. coli isolate. Such surface exposure has implications for the use of these Mabs for APEC detection and colibacillosis control.

A Two-Stage Reconstruction Processor for Human Detection in Compressive Sensing CMOS Radar.

PubMed

Tsao, Kuei-Chi; Lee, Ling; Chu, Ta-Shun; Huang, Yuan-Hao

2018-04-05

Complementary metal-oxide-semiconductor (CMOS) radar has recently gained much research attraction because small and low-power CMOS devices are very suitable for deploying sensing nodes in a low-power wireless sensing system. This study focuses on the signal processing of a wireless CMOS impulse radar system that can detect humans and objects in the home-care internet-of-things sensing system. The challenges of low-power CMOS radar systems are the weakness of human signals and the high computational complexity of the target detection algorithm. The compressive sensing-based detection algorithm can relax the computational costs by avoiding the utilization of matched filters and reducing the analog-to-digital converter bandwidth requirement. The orthogonal matching pursuit (OMP) is one of the popular signal reconstruction algorithms for compressive sensing radar; however, the complexity is still very high because the high resolution of human respiration leads to high-dimension signal reconstruction. Thus, this paper proposes a two-stage reconstruction algorithm for compressive sensing radar. The proposed algorithm not only has lower complexity than the OMP algorithm by 75% but also achieves better positioning performance than the OMP algorithm especially in noisy environments. This study also designed and implemented the algorithm by using Vertex-7 FPGA chip (Xilinx, San Jose, CA, USA). The proposed reconstruction processor can support the 256 × 13 real-time radar image display with a throughput of 28.2 frames per second.

Experimental investigations on airborne gravimetry based on compressed sensing.

PubMed

Yang, Yapeng; Wu, Meiping; Wang, Jinling; Zhang, Kaidong; Cao, Juliang; Cai, Shaokun

2014-03-18

Gravity surveys are an important research topic in geophysics and geodynamics. This paper investigates a method for high accuracy large scale gravity anomaly data reconstruction. Based on the airborne gravimetry technology, a flight test was carried out in China with the strap-down airborne gravimeter (SGA-WZ) developed by the Laboratory of Inertial Technology of the National University of Defense Technology. Taking into account the sparsity of airborne gravimetry by the discrete Fourier transform (DFT), this paper proposes a method for gravity anomaly data reconstruction using the theory of compressed sensing (CS). The gravity anomaly data reconstruction is an ill-posed inverse problem, which can be transformed into a sparse optimization problem. This paper uses the zero-norm as the objective function and presents a greedy algorithm called Orthogonal Matching Pursuit (OMP) to solve the corresponding minimization problem. The test results have revealed that the compressed sampling rate is approximately 14%, the standard deviation of the reconstruction error by OMP is 0.03 mGal and the signal-to-noise ratio (SNR) is 56.48 dB. In contrast, the standard deviation of the reconstruction error by the existing nearest-interpolation method (NIPM) is 0.15 mGal and the SNR is 42.29 dB. These results have shown that the OMP algorithm can reconstruct the gravity anomaly data with higher accuracy and fewer measurements.

Experimental Investigations on Airborne Gravimetry Based on Compressed Sensing

PubMed Central

Yang, Yapeng; Wu, Meiping; Wang, Jinling; Zhang, Kaidong; Cao, Juliang; Cai, Shaokun

2014-01-01

Gravity surveys are an important research topic in geophysics and geodynamics. This paper investigates a method for high accuracy large scale gravity anomaly data reconstruction. Based on the airborne gravimetry technology, a flight test was carried out in China with the strap-down airborne gravimeter (SGA-WZ) developed by the Laboratory of Inertial Technology of the National University of Defense Technology. Taking into account the sparsity of airborne gravimetry by the discrete Fourier transform (DFT), this paper proposes a method for gravity anomaly data reconstruction using the theory of compressed sensing (CS). The gravity anomaly data reconstruction is an ill-posed inverse problem, which can be transformed into a sparse optimization problem. This paper uses the zero-norm as the objective function and presents a greedy algorithm called Orthogonal Matching Pursuit (OMP) to solve the corresponding minimization problem. The test results have revealed that the compressed sampling rate is approximately 14%, the standard deviation of the reconstruction error by OMP is 0.03 mGal and the signal-to-noise ratio (SNR) is 56.48 dB. In contrast, the standard deviation of the reconstruction error by the existing nearest-interpolation method (NIPM) is 0.15 mGal and the SNR is 42.29 dB. These results have shown that the OMP algorithm can reconstruct the gravity anomaly data with higher accuracy and fewer measurements. PMID:24647125

Evaluation of immunogenicity and protective efficacy of recombinant outer membrane proteins of Haemophilus parasuis serovar 5 in a murine model.

PubMed

Li, Miao; Cai, Ru-Jian; Song, Shuai; Jiang, Zhi-Yong; Li, Yan; Gou, Hong-Chao; Chu, Pin-Pin; Li, Chun-Ling; Qiu, Hua-Ji

2017-01-01

Glässer's disease is an economically important infectious disease of pigs caused by Haemophilus parasuis. Few vaccines are currently available that could provide effective cross-protection against various serovars of H. parasuis. In this study, five OMPs (OppA, TolC, HxuC, LppC, and HAPS_0926) identified by bioinformatic approaches, were cloned and expressed as recombinant proteins. Antigenicity of the purified proteins was verified through Western blotting, and primary screening for protective potential was evaluated in vivo. Recombinant TolC (rTolC), rLppC, and rHAPS_0926 proteins showing marked protection of mice against H. parasuis infection, and were further evaluated individually or in combination. Mice treated with these three OMPs produced humoral and host cell-mediated responses, with a significant rise in antigen-specific IgG titer and lymphoproliferative response in contrast with the mock-immunized group. Significant increases were noted in CD4+, CD8+ T cells, and three cytokines (IL-2, IL-4, and IFN-γ) in vaccinated animals. The antisera against candidate antigens could efficiently impede bacterial survival in whole blood bactericidal assay against H. parasuis infection. The multi-protein vaccine induced more pronounced immune responses and offered better protection than individual vaccines. Our findings indicate that these three OMPs are promising antigens for the development of multi-component subunit vaccines against Glässer's disease.

Data Continuity of Aerosol Index from Suomi NPP/OMPS Observations

NASA Astrophysics Data System (ADS)

Ahn, C.; Torres, O.; Tiruchirapalli, R.; Taylor, S.; Jethva, H. T.

2017-12-01

Since the development of the Aerosol Index (AI) concept from Nimubs-7 TOMS near-UV measurements, the AI product has been widely used by the aerosol community in a variety of applications including monitoring of the sources and sinks of carbonaceous and desert dust aerosols. The AI uses a pair of near-UV radiances to detect the presence of absorbing particles even over bright backgrounds such as clouds and snow/ice covered areas. Since its inception in the mid 90's, the AI has been available as a by-product of the total ozone product. Due to the implementation of a new total ozone algorithm, the standard AI product will no longer be available starting in 2018. To assure the continuity of the AI record, we have developed an improved AI algorithm that uses a better forward modeling method of the top of atmosphere radiances. The enhanced modelling capability accounts for the scattering of clouds using Mie theory, and includes the effect of wavelength and angle dependent surface reflectance effects. By doing this, we have significantly reduced angular dependent false AI signals such as sun glint over the ocean. We will discuss the improved AI algorithm and present the long term AI record from various UV space borne sensors including TOMS, OMI, OMPS, and EPIC with consistent AI algorithms, followed by future plans for near-real time processing and operational production of a new OMPS AI product.

Synthesis of an oligonucleotide-derivatized amphipol and its use to trap and immobilize membrane proteins

PubMed Central

Bon, Christel Le; Della Pia, Eduardo Antonio; Giusti, Fabrice; Lloret, Noémie; Zoonens, Manuela; Martinez, Karen L.; Popot, Jean-Luc

2014-01-01

Amphipols (APols) are specially designed amphipathic polymers that stabilize membrane proteins (MPs) in aqueous solutions in the absence of detergent. A8–35, a polyacrylate-based APol, has been grafted with an oligodeoxynucleotide (ODN). The synthesis, purification and properties of the resulting ‘OligAPol’ have been investigated. Grafting was performed by reacting an ODN carrying an amine-terminated arm with the carboxylates of A8–35. The use of OligAPol for trapping MPs and immobilizing them onto solid supports was tested using bacteriorhodopsin (BR) and the transmembrane domain of Escherichia coli outer membrane protein A (tOmpA) as model proteins. BR and OligAPol form water-soluble complexes in which BR remains in its native conformation. Hybridization of the ODN arm with a complementary ODN was not hindered by the assembly of OligAPol into particles, nor by its association with BR. BR/OligAPol and tOmpA/OligAPol complexes could be immobilized onto either magnetic beads or gold nanoparticles grafted with the complementary ODN, as shown by spectroscopic measurements, fluorescence microscopy and the binding of anti-BR and anti-tOmpA antibodies. OligAPols provide a novel, highly versatile approach to tagging MPs, without modifying them chemically nor genetically, for specific, reversible and targetable immobilization, e.g. for nanoscale applications. PMID:24744236

Simultaneous determination of 17 sulfonamides and the potentiators ormetoprim and trimethoprim in salmon muscle by liquid chromatography with tandem mass spectrometry detection.

PubMed

Potter, Ross A; Burns, B Garth; van de Riet, Jeffrey M; North, David H; Darvesh, Rozina

2007-01-01

A simple, robust method using liquid chromatography/tandem mass spectrometry (LC/MS/MS) for the simultaneous determination of 17 sulfonamides [sulfanilamide (SNL), sulfacetamide (SAA), sulfaguanidine (SGD), sulfapyridine (SPY), sulfadiazine (SDZ), sulfathiazole (STZ), sulfamerazine (SMR), sulfamethoxazole (SOZ), sulfamoxole (SXL), sulfisoxazole (SXZ), sulfamethizole (SML), sulfamethazine (SMZ), sulfamethoxypyridazine (SMP), sulfamonomethoxine (SMM), sulfachloropyridazine (SCP), sulfaquinoxaline (SQX), and sulfadimethoxine (SDM)] and 2 potentiators [ormetoprim (OMP) and trimethoprim (TMP)] in fish tissue has been developed. The analytes were extracted from homogenized fish tissue with water-acetonitrile (50 + 50). The extract was clarified by centrifugation and a portion defatted with hexane. The analytes were partitioned into chloroform and evaporated to dryness. The redissolved residue was applied to a C18 reversed-phase column with a water-acetonitrile (0.1% acetic acid) gradient. All of the compounds were completely separated and detected in <10 min at 30 degrees C using LC/MS/MS. Standard curves were linear over the range of 0.02 to 5 ng injected. The limit of detection varied from 0.1 ng/g for SMZ and OMP to 0.9 ng/g for SXL and SOZ. Recoveries varied from 100% for SDM, SOZ, and SQX and 85% for SMR, OMP, and TMP to approximately 30% for SAA. Relative standard deviations for repeat analysis varied from 4% for SMZ and SCP to 23% for SAA.

Marginalized Student Access to Technology Education

NASA Astrophysics Data System (ADS)

Kurtcu, Wanda M.

The purpose of this paper is to investigate how a teacher can disrupt an established curriculum that continues the cycle of inequity of access to science, technology, engineering, and math (STEM) curriculum by students in alternative education. For this paper, I will focus on the technology components of the STEM curriculum. Technology in the United States, if not the world economy, is developing at a rapid pace. Many areas of day to day living, from applying for a job to checking one's bank account online, involve a component of science and technology. The 'gap' in technology education is emphasized between the 'haves and have-nots', which is delineated along socio-economic lines. Marginalized students in alternative education programs use this equipment for little else than remedial programs and credit recovery. This level of inequity further widens in alternative education programs and affects the achievement of marginalized students in credit recovery or alternative education classes instead of participation technology classes. For the purposes of this paper I focus on how can I decrease the inequity of student access to 21st century technology education in an alternative education program by addressing the established curriculum of the program and modifying structural barriers of marginalized student access to a technology focused curriculum.

Presurgical language fMRI: Clinical practices and patient outcomes in epilepsy surgical planning.

PubMed

Benjamin, Christopher F A; Li, Alexa X; Blumenfeld, Hal; Constable, R Todd; Alkawadri, Rafeed; Bickel, Stephan; Helmstaedter, Christoph; Meletti, Stefano; Bronen, Richard; Warfield, Simon K; Peters, Jurriaan M; Reutens, David; Połczyńska, Monika; Spencer, Dennis D; Hirsch, Lawrence J

2018-03-12

The goal of this study was to document current clinical practice and report patient outcomes in presurgical language functional MRI (fMRI) for epilepsy surgery. Epilepsy surgical programs worldwide were surveyed as to the utility, implementation, and efficacy of language fMRI in the clinic; 82 programs responded. Respondents were predominantly US (61%) academic programs (85%), and evaluated adults (44%), adults and children (40%), or children only (16%). Nearly all (96%) reported using language fMRI. Surprisingly, fMRI is used to guide surgical margins (44% of programs) as well as lateralize language (100%). Sites using fMRI for localization most often use a distance margin around activation of 10mm. While considered useful, 56% of programs reported at least one instance of disagreement with other measures. Direct brain stimulation typically confirmed fMRI findings (74%) when guiding margins, but instances of unpredicted decline were reported by 17% of programs and 54% reported unexpected preservation of function. Programs reporting unexpected decline did not clearly differ from those which did not. Clinicians using fMRI to guide surgical margins do not typically map known language-critical areas beyond Broca's and Wernicke's. This initial data shows many clinical teams are confident using fMRI not only for language lateralization but also to guide surgical margins. Reported cases of unexpected language preservation when fMRI activation is resected, and cases of language decline when it is not, emphasize a critical need for further validation. Comprehensive studies comparing commonly-used fMRI paradigms to predict stimulation mapping and post-surgical language decline remain of high importance. © 2018 The Authors Human Brain Mapping Published by Wiley Periodicals, Inc.

Access and Retention of Marginalized Populations within Undergraduate Music Education Degree Programs

ERIC Educational Resources Information Center

Fitzpatrick, Kate R.; Henninger, Jacqueline C.; Taylor, Don M.

2014-01-01

The purpose of this collective instrumental case study was to examine the experiences of six undergraduate students from traditionally marginalized populations with regard to their preparation for, admission to, and retention within a music education degree program. Analyzed and reported through the lens of critical theory, data sources included…

12 CFR 221.122 - Applicability of margin requirements to credit in connection with Insurance Premium Funding...

Code of Federal Regulations, 2011 CFR

2011-01-01

... in connection with Insurance Premium Funding Programs. 221.122 Section 221.122 Banks and Banking...) Interpretations § 221.122 Applicability of margin requirements to credit in connection with Insurance Premium Funding Programs. (a) The Board has been asked numerous questions regarding purpose credit in connection...

12 CFR 221.122 - Applicability of margin requirements to credit in connection with Insurance Premium Funding...

Code of Federal Regulations, 2010 CFR

2010-01-01

... in connection with Insurance Premium Funding Programs. 221.122 Section 221.122 Banks and Banking...) Interpretations § 221.122 Applicability of margin requirements to credit in connection with Insurance Premium Funding Programs. (a) The Board has been asked numerous questions regarding purpose credit in connection...

Differential In Vivo Gene Expression of Major Leptospira Proteins in Resistant or Susceptible Animal Models

PubMed Central

Matsui, Mariko; Soupé, Marie-Estelle; Becam, Jérôme

2012-01-01

Transcripts of Leptospira 16S rRNA, FlaB, LigB, LipL21, LipL32, LipL36, LipL41, and OmpL37 were quantified in the blood of susceptible (hamsters) and resistant (mice) animal models of leptospirosis. We first validated adequate reference genes and then evaluated expression patterns in vivo compared to in vitro cultures. LipL32 expression was downregulated in vivo and differentially regulated in resistant and susceptible animals. FlaB expression was also repressed in mice but not in hamsters. In contrast, LigB and OmpL37 were upregulated in vivo. Thus, we demonstrated that a virulent strain of Leptospira differentially adapts its gene expression in the blood of infected animals. PMID:22729538

Identification and strain differentiation of Vibrio cholerae by using polyclonal antibodies against outer membrane proteins.

PubMed

Martínez-Govea, A; Ambrosio, J; Gutiérrez-Cogco, L; Flisser, A

2001-07-01

Cholera is caused only by O1 and O139 Vibrio cholerae strains. For diagnosis, 3 working days are needed for bacterial isolation from human feces and for biochemical characterization. Here we describe the purification of bacterial outer membrane proteins (OMP) from V. cholerae O1 Ogawa, O1 Inaba, and O139 strains, as well as the production of specific antisera and their use for fecal Vibrio antigen detection. Anti-OMP antisera showed very high reactivity and specificity by enzyme-linked immunosorbent assay (ELISA) and dot-ELISA. An inmunodiagnostic assay for V. cholerae detection was developed; this assay avoids preenrichment and costly equipment and can be used for epidemiological surveillance and clinical diagnosis of cases, considering that prompt and specific identification of bacteria is mandatory in cholera.

A novel aliasing-free subband information fusion approach for wideband sparse spectral estimation

NASA Astrophysics Data System (ADS)

Luo, Ji-An; Zhang, Xiao-Ping; Wang, Zhi

2017-12-01

Wideband sparse spectral estimation is generally formulated as a multi-dictionary/multi-measurement (MD/MM) problem which can be solved by using group sparsity techniques. In this paper, the MD/MM problem is reformulated as a single sparse indicative vector (SIV) recovery problem at the cost of introducing an additional system error. Thus, the number of unknowns is reduced greatly. We show that the system error can be neglected under certain conditions. We then present a new subband information fusion (SIF) method to estimate the SIV by jointly utilizing all the frequency bins. With orthogonal matching pursuit (OMP) leveraging the binary property of SIV's components, we develop a SIF-OMP algorithm to reconstruct the SIV. The numerical simulations demonstrate the performance of the proposed method.

Prokaryotic 2-component systems and the OmpR/PhoB superfamily.

PubMed

Nguyen, Minh-Phuong; Yoon, Joo-Mi; Cho, Man-Ho; Lee, Sang-Won

2015-11-01

In bacteria, 2-component regulatory systems (TCSs) are the critical information-processing pathways that link stimuli to specific adaptive responses. Signals perceived by membrane sensors, which are generally histidine kinases, are transmitted by response regulators (RRs) to allow cells to cope rapidly and effectively with environmental challenges. Over the past few decades, genes encoding components of TCSs and their responsive proteins have been identified, crystal structures have been described, and signaling mechanisms have been elucidated. Here, we review recent findings and interesting breakthroughs in bacterial TCS research. Furthermore, we discuss structural features, mechanisms of activation and regulation, and cross-regulation of RRs, with a focus on the largest RR family, OmpR/PhoB, to provide a comprehensive overview of these critically important signaling molecules.

Organic micropollutants (OMPs) in natural waters: Oxidation by UV/H2O2 treatment and toxicity assessment.

PubMed

Rozas, Oscar; Vidal, Cristiane; Baeza, Carolina; Jardim, Wilson F; Rossner, Alfred; Mansilla, Héctor D

2016-07-01

Organic micropollutants (OMPs) are ubiquitous in natural waters even in places where the human activity is limited. The presence of OMPs in natural water sources for human consumption encourages the evaluation of different water purification technologies to ensure water quality. In this study, the Biobío river (Chile) was selected since the watershed includes urban settlements and economic activities (i.e. agriculture, forestry) that incorporate a variety of OMPs into the aquatic environment, such as pesticides, pharmaceuticals and personal care products. Atrazine (herbicide), caffeine (psychotropic), diclofenac (anti-inflammatory) and triclosan (antimicrobial) in Biobío river water and in different stages of a drinking and two wastewater treatment plants downstream Biobío river were determined using solid phase extraction (SPE) and liquid chromatography/tandem mass spectrometry (LC-MS/MS) and electrospray ionization (ESI). Quantification of these four compounds showed concentrations in the range of 8 ± 2 to 55 ± 10 ng L(-1) in Biobío river water, 11 ± 2 to 74 ± 21 ng L(-1) in the drinking water treatment plant, and 60 ± 10 to 15,000 ± 1300 ng L(-1) in the wastewater treatment plants. Caffeine was used as an indicator of wastewater discharges. Because conventional water treatment technologies are not designed to eliminate some emerging organic pollutants, alternative treatment processes, UV and UV/H2O2, were employed. The transformation of atrazine, carbamazepine (antiepileptic), diclofenac and triclosan was investigated at laboratory scale. Both processes were tested at different UV doses and the Biobío river water matrix effects were evaluated. Initial H2O2 concentration used was 10 mg L(-1). Results showed that, the transformation profile obtained using UV/H2O2 at UV doses up to 900 mJ cm(-2), followed the trend of diclofenac > triclosan > atrazine > carbamazepine. Furthermore acute toxicity tests with Daphnia magna were carried out after UV/H2O2 treatments of the OMPs mixture studied. At the lower UV doses tested (300 mJ cm(-2)) a higher toxicity was observed, suggesting the formation of toxic intermediates in the course of the reaction. As expected, at higher UV doses the toxicity declined. Considering the treatment of the mixture of ATZ, CBZ, DCL and TCS with a UV dose of 1200 mJ cm(-2) and 10 mg L(-1) of H2O2 the acute toxicity results exhibits values for Daphnia magna immobilization equal to 20 and 42% evaluated after 24 and 48 h, respectively. Copyright © 2016 Elsevier Ltd. All rights reserved.

Supramolecular architectures in Co(II) and Cu(II) complexes with thiophene-2-carboxylate and 2-amino-4,6-dimethoxypyrimidine ligands.

PubMed

Karthikeyan, Ammasai; Thomas Muthiah, Packianathan; Perdih, Franc

2016-05-01

The coordination chemistry of mixed-ligand complexes continues to be an active area of research since these compounds have a wide range of applications. Many coordination polymers and metal-organic framworks are emerging as novel functional materials. Aminopyrimidine and its derivatives are flexible ligands with versatile binding and coordination modes which have been proven to be useful in the construction of organic-inorganic hybrid materials and coordination polymers. Thiophenecarboxylic acid, its derivatives and their complexes exhibit pharmacological properties. Cobalt(II) and copper(II) complexes of thiophenecarboxylate have many biological applications, for example, as antifungal and antitumor agents. Two new cobalt(II) and copper(II) complexes incorporating thiophene-2-carboxylate (2-TPC) and 2-amino-4,6-dimethoxypyrimidine (OMP) ligands have been synthesized and characterized by X-ray diffraction studies, namely (2-amino-4,6-dimethoxypyrimidine-κN)aquachlorido(thiophene-2-carboxylato-κO)cobalt(II) monohydrate, [Co(C5H3O2S)Cl(C6H9N3O2)(H2O)]·H2O, (I), and catena-poly[copper(II)-tetrakis(μ-thiophene-2-carboxylato-κ(2)O:O')-copper(II)-(μ-2-amino-4,6-dimethoxypyrimidine-κ(2)N(1):N(3))], [Cu2(C5H3O2S)4(C6H9N3O2)]n, (II). In (I), the Co(II) ion has a distorted tetrahedral coordination environment involving one O atom from a monodentate 2-TPC ligand, one N atom from an OMP ligand, one chloride ligand and one O atom of a water molecule. An additional water molecule is present in the asymmetric unit. The amino group of the coordinated OMP molecule and the coordinated carboxylate O atom of the 2-TPC ligand form an interligand N-H...O hydrogen bond, generating an S(6) ring motif. The pyrimidine molecules also form a base pair [R2(2)(8) motif] via a pair of N-H...N hydrogen bonds. These interactions, together with O-H...O and O-H...Cl hydrogen bonds and π-π stacking interactions, generate a three-dimensional supramolecular architecture. The one-dimensional coordination polymer (II) contains the classical paddle-wheel [Cu2(CH3COO)4(H2O)2] unit, where each carboxylate group of four 2-TPC ligands bridges two square-pyramidally coordinated Cu(II) ions and the apically coordinated OMP ligands bridge the dinuclear copper units. Each dinuclear copper unit has a crystallographic inversion centre, whereas the bridging OMP ligand has crystallographic twofold symmetry. The one-dimensional polymeric chains self-assemble via N-H...O, π-π and C-H...π interactions, generating a three-dimensional supramolecular architecture.

A high-resolution conceptual model for diffuse organic micropollutant loads in streams

NASA Astrophysics Data System (ADS)

Stamm, Christian; Honti, Mark; Ghielmetti, Nico

2013-04-01

The ecological state of surface waters has become the dominant aspect in water quality assessments. Toxicity is a key determinant of the ecological state, but organic micropollutants (OMP) are seldom monitored with the same spatial and temporal frequency as for example nutrients, mainly due the demanding analytical methods and costs. However, diffuse transport pathways are at least equally complex for OMPs as for nutrients and there are still significant knowledge gaps. Moreover, concentrations of the different compounds would need to be known with fairly high temporal resolution because acute toxicity can be as important as the chronic one. Fully detailed mechanistic models of diffuse OMP loads require an immense set of site-specific knowledge and are rarely applicable for catchments lacking an exceptional monitoring coverage. Simple empirical methods are less demanding but usually work with more temporal aggregation and that's why they have limited possibilities to support the estimation of the ecological state. This study presents a simple conceptual model that aims to simulate the concentrations of selected organic micropollutants with daily resolution at 11 locations in the stream network of a small catchment (46 km2). The prerequisite is a known hydrological and meteorological background (daily discharge, precipitation and air temperature time series), a land use map and some historic measurements of the desired compounds. The model is conceptual in the sense that all important diffuse transport pathways are simulated separately, but each with a simple empirical process rate. Consequently, some site-specific observations are required to calibrate the model, but afterwards the model can be used for forecasting and scenario analysis as the calibrated process rates typically describe invariant properties of the catchment. We simulated 6 different OMPs from the categories of agricultural and urban pesticides and urban biocides. The application of agricultural pesticides was also simulated with the model using a heat-sum approach. Calibration was carried out with weekly aggregated samples covering the growing season in 2 years. The model could reproduce the observed OMP concentrations with varying success. Compounds that are less persistent in the environment and thus have a dominant temporal dynamics (pesticides with a short half-life) could be simulated in general better than the persistent ones. For the latter group the relatively stable available stock meant that there were no clear seasonal dynamics, which revealed that transport processes are quite uncertain even when daily rainfall is used as the main driver. Nevertheless the daily concentration distribution could still be simulated with higher accuracy than the individual peaks. Thus we can model the concentration-duration relationship for daily resolution in an acceptable way for each compound.

76 FR 19512 - Self-Regulatory Organizations; The Options Clearing Corporation; Notice of Filing of Proposed...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-04-07

... clearing organizations. In the internal cross-margining program, there is no need for two separate accounts... Organizations; The Options Clearing Corporation; Notice of Filing of Proposed Rule Change to Allow for an Expansion of OCC's Internal Cross-Margining Program to Include the Ability of a Pair of Affiliated Clearing...

Fulfilling Their Dreams: Marginalized Urban Youths' Perspectives on a Culturally Sensitive Social and Emotional Learning Program

ERIC Educational Resources Information Center

Slaten, Christopher D.; Rivera, Roberto C.; Shemwell, Daniel; Elison, Zachary M.

2016-01-01

A growing body of research suggests educators need to focus on cultivating social and emotional competencies that youth will need to thrive in the new knowledge economy (Durlak, Weissberg, Dymnicki, Taylor, & Schellinger, 2011). For marginalized urban youth, in particular, few have derived programs and interventions to assist with these…

Antibodies to CBir1 are Associated with Glycogen Storage Disease Type Ib

PubMed Central

Davis, Michael K.; Valentine, John F.; Weinstein, David A.; Polyak, Steven

2009-01-01

Objectives Glycogen storage disease (GSD) type Ib is a congenital disorder of glycogen metabolism that is associated with neutropenia, neutrophil and monocyte dysfunction, and an inflammatory bowel disease (IBD) that mimics a Crohn's disease phenotype. The enteric microflora is implicated in the pathogenesis of IBD; however, its role in the development of GSD-associated IBD is unknown. Antibody reactivity to Saccharomyces cerevisiae antibodies (ASCA), Escherichia coli outer membrane porin C (anti-OmpC) and bacterial flagellin (anti-CBir1) have been associated with Crohn's disease in the general population, but they have an undetermined association in children and adults with GSD-Ib. Our goal was to examine the association of ASCA, anti-OmpC and anti-CBir1 with the clinical features of GSD-Ib enterocolitis. Methods A retrospective review identified 19 GSD-Ib patients with or without a known diagnosis of enterocolitis. Radiographic, endoscopic and serologic data were collected and assays for ASCA, anti-OmpC, and anti-CBir1 obtained. Results Seven patients had combined radiographic, endoscopic, and histological evidence of intestinal inflammation; the majority had ileocolonic involvement. Seventeen of 19 (89%) patients had elevated anti-CBir1 levels (6/7 in the IBD group and 11/12 in the no clinical evidence of IBD group). Thirteen of 19 (68%) had elevated anti-OmpC levels (5/7 in the IBD group and 8/12 in the no clinical evidence of IBD group). Eleven of 19 (58%) patients had elevated ASCA IgA levels (4/7 in the IBD group and 7/12 in the no clinical evidence of IBD group). Conclusion Nearly all of the GSD type Ib patients had elevated anti-CBir1 levels. The antibody did not differentiate those with and without a diagnosis of GSD-Ib-associated IBD. Seroreactivity to flagellin may represent immune dysfunction rather than active enterocolitis in this patient population. Long-term follow-up of the group without known IBD is required to determine if these antibodies can predict intestinal inflammation. PMID:20410847

Serum antibody response to Moraxella catarrhalis proteins OMP CD, OppA, Msp22, Hag, and PilA2 after nasopharyngeal colonization and acute otitis media in children.

PubMed

Ren, Dabin; Almudevar, Anthony L; Murphy, Timothy F; Lafontaine, Eric R; Campagnari, Anthony A; Luke-Marshall, Nicole; Casey, Janet R; Pichichero, Michael E

2015-10-26

There is no licensed vaccine for Moraxella catarrhalis (Mcat), which is a prominent bacterium causing acute otitis media (AOM) in children and lower respiratory tract infections in adults. Nasopharyngeal (NP) colonization caused by respiratory bacteria results in natural immunization of the host. To identify Mcat antigens as vaccine candidates, we evaluated the development of naturally induced antibodies to 5 Mcat surface proteins in children 6-30 months of age during Mcat NP colonization and AOM. Human serum IgG against the recombinant Mcat proteins, outer membrane protein (OMP) CD, oligopeptide permease (Opp)A, hemagglutinin (Hag), Moraxella surface protein (Msp)22, and PilA clade 2 (PilA2) was quantitated by using an ELISA assay. There were 223 Mcat NP colonization episodes documented in 111 (60%) of 184 children in the study. Thirty five Mcat AOM episodes occurred in 30 (16%) of 184 children. All 5 Mcat candidate vaccine antigens evaluated stimulated a significant rise in serum IgG levles over time from 6 to 36 months of age (P<0.001), with a rank order as follows: Msp22=OppA>OMP CD=Hag=PilA2. Children with no detectable Mcat NP colonization showed a higher serum IgG level against OppA, Hag, and Msp22 compared to those with Mcat NP colonization (P<0.05). Individual data showed that some children responded to AOM with an antibody increase to one or more of the studied Mcat proteins but some children failed to respond. Serum antibody to Mcat candidate vaccine proteins OMP CD, OppA, Msp22, Hag, and PilA2 increased with age in naturally immunized children age 6-30 months following Mcat NP colonization and AOM. High antibody levels against OppA, Msp22, and Hag correlated with reduced carriage. The results support further investigation of these vaccine candidates in protecting against Mcat colonization and infection. Copyright © 2015 Elsevier Ltd. All rights reserved.

Stringently Defined Otitis Prone Children Demonstrate Deficient Naturally Induced Mucosal Antibody Response to Moraxella catarrhalis Proteins

PubMed Central

Ren, Dabin; Murphy, Timothy F.; Lafontaine, Eric R.; Pichichero, Michael E.

2017-01-01

Moraxella catarrhalis (Mcat) is a prominent mucosal pathogen causing acute otitis media (AOM). We studied Mcat nasopharyngeal (NP) colonization, AOM frequency and mucosal antibody responses to four vaccine candidate Mcat proteins: outer membrane protein (OMP) CD, oligopeptide permease (Opp) A, hemagglutinin (Hag), and Pilin A clade 2 (PilA2) from stringently defined otitis prone (sOP) children, who experience the greatest burden of disease, compared to non-otitis prone (NOP) children. sOP children had higher NP colonization of Mcat (30 vs. 22%, P = 0.0003) and Mcat-caused AOM rates (49 vs. 24%, P < 0.0001) than NOP children. Natural acquisition of mucosal antibodies to Mcat proteins OMP CD (IgG, P < 0.0001), OppA (IgG, P = 0.018), Hag (IgG and IgA, both P < 0.0001), and PilA2 (IgA, P < 0.0001) was lower in sOP than NOP children. Higher levels of mucosal IgG to Hag (P = 0.039) and PilA2 (P = 0.0076), and IgA to OMP CD (P = 0.010), OppA (P = 0.030), and PilA2 (P = 0.043) were associated with lower carriage of Mcat in NOP but not sOP children. Higher levels of mucosal IgG to OMP CD (P = 0.0070) and Hag (P = 0.0003), and IgA to Hag (P = 0.0067) at asymptomatic colonization than those at onset of AOM were associated with significantly lower rate of Mcat NP colonization progressing to AOM in NOP compared to sOP children (3 vs. 26%, P < 0.0001). In conclusion, sOP children had a diminished mucosal antibody response to Mcat proteins, which was associated with higher frequencies of asymptomatic NP colonization and NP colonization progressing to Mcat-caused AOM. Enhancing Mcat antigen-specific mucosal immune responses to levels higher than achieved by natural exposure will be necessary to prevent AOM in sOP children. PMID:28848555

Stringently Defined Otitis Prone Children Demonstrate Deficient Naturally Induced Mucosal Antibody Response to Moraxella catarrhalis Proteins.

PubMed

Ren, Dabin; Murphy, Timothy F; Lafontaine, Eric R; Pichichero, Michael E

2017-01-01

Moraxella catarrhalis ( Mcat ) is a prominent mucosal pathogen causing acute otitis media (AOM). We studied Mcat nasopharyngeal (NP) colonization, AOM frequency and mucosal antibody responses to four vaccine candidate Mcat proteins: outer membrane protein (OMP) CD, oligopeptide permease (Opp) A, hemagglutinin (Hag), and Pilin A clade 2 (PilA2) from stringently defined otitis prone (sOP) children, who experience the greatest burden of disease, compared to non-otitis prone (NOP) children. sOP children had higher NP colonization of Mcat (30 vs. 22%, P  = 0.0003) and Mcat -caused AOM rates (49 vs. 24%, P  < 0.0001) than NOP children. Natural acquisition of mucosal antibodies to Mcat proteins OMP CD (IgG, P  < 0.0001), OppA (IgG, P  = 0.018), Hag (IgG and IgA, both P  < 0.0001), and PilA2 (IgA, P  < 0.0001) was lower in sOP than NOP children. Higher levels of mucosal IgG to Hag ( P  = 0.039) and PilA2 ( P  = 0.0076), and IgA to OMP CD ( P  = 0.010), OppA ( P  = 0.030), and PilA2 ( P  = 0.043) were associated with lower carriage of Mcat in NOP but not sOP children. Higher levels of mucosal IgG to OMP CD ( P  = 0.0070) and Hag ( P  = 0.0003), and IgA to Hag ( P  = 0.0067) at asymptomatic colonization than those at onset of AOM were associated with significantly lower rate of Mcat NP colonization progressing to AOM in NOP compared to sOP children (3 vs. 26%, P  < 0.0001). In conclusion, sOP children had a diminished mucosal antibody response to Mcat proteins, which was associated with higher frequencies of asymptomatic NP colonization and NP colonization progressing to Mcat -caused AOM. Enhancing Mcat antigen-specific mucosal immune responses to levels higher than achieved by natural exposure will be necessary to prevent AOM in sOP children.

Serum antibody response to Moraxella catarrhalis proteins OMP CD, OppA, Msp22, Hag, and PilA2 after nasopharyngeal colonization and acute otitis media in children

PubMed Central

Ren, Dabin; Almudevar, Anthony L.; Murphy, Timothy F.; Lafontaine, Eric R.; Campagnari, Anthony A.; Luke-Marshall, Nicole; Casey, Janet R.; Pichichero, Michael E.

2015-01-01

Background There is no licensed vaccine for Moraxella catarrhalis (Mcat), which is a prominent bacterium causing acute otitis media (AOM) in children and lower respiratory tract infections in adults. Nasopharyngeal (NP) colonization caused by respiratory bacteria results in natural immunization of the host. To identify Mcat antigens as vaccine candidates, we evaluated the development of naturally induced antibodies to 5 Mcat surface proteins in children 6–30 months of age during Mcat NP colonization and AOM. Methods Human serum IgG against the recombinant Mcat proteins, outer membrane protein (OMP) CD, oligopeptide permease (Opp)A, hemagglutinin (Hag), Moraxella surface protein (Msp)22, and PilA clade 2 (PilA2) was quantitated by using an ELISA assay. Results There were 223 Mcat NP colonization episodes documented in 111 (60%) of 184 children in the study. Thirty five Mcat AOM episodes occurred in 30 (16%) of 184 children. All 5 Mcat candidate vaccine antigens evaluated stimulated a significant rise in serum IgG levles over time from 6 to 36 months of age (P < 0.001), with a rank order as follows: Msp22 = OppA > OMP CD = Hag = PilA2. Children with no detectable Mcat NP colonization showed a higher serum IgG level against OppA, Hag, and Msp22 compared to those with Mcat NP colonization (P < 0.05). Individual data showed that some children responded to AOM with an antibody increase to one or more of the studied Mcat proteins but some children failed to respond. Conclusions Serum antibody to Mcat candidate vaccine proteins OMP CD, OppA, Msp22, Hag, and PilA2 increased with age in naturally immunized children age 6–30 months following Mcat NP colonization and AOM. High antibody levels against OppA, Msp22, and Hag correlated with reduced carriage. The results support further investigation of these vaccine candidates in protecting against Mcat colonization and infection. PMID:26392013

Immunogenicity of 2 serogroup B outer-membrane protein meningococcal vaccines: a randomized controlled trial in Chile.

PubMed

Tappero, J W; Lagos, R; Ballesteros, A M; Plikaytis, B; Williams, D; Dykes, J; Gheesling, L L; Carlone, G M; Høiby, E A; Holst, J; Nøkleby, H; Rosenqvist, E; Sierra, G; Campa, C; Sotolongo, F; Vega, J; Garcia, J; Herrera, P; Poolman, J T; Perkins, B A

1999-04-28

Meningococcal disease occurs worldwide, and serogroup B disease accounts for a large proportion of cases. Although persons younger than 4 years are at greatest risk for serogroup B meningococcal disease, vaccine efficacy has not been demonstrated in this age group. To evaluate serum bactericidal activity (SBA) against homologous vaccine type strains and a heterologous Chilean epidemic strain of Neisseria meningitidis as a potential correlate for vaccine efficacy. Double-blind, randomized controlled trial conducted between March 14 and July 20, 1994. All blood samples were taken by December 1994. Santiago, Chile, where a clonal serogroup B meningococcal disease epidemic began in 1993. Infants younger than 1 year (n = 187), children aged 2 to 4 years (n = 183), and adults aged 17 to 30 years (n = 173). Participants received 3 doses of outer-membrane protein (OMP) meningococcal vaccine developed in either Cuba or Norway or a control vaccine, with each dose given 2 months apart. Blood samples were obtained at baseline, prior to dose 3, and at 4 to 6 weeks after dose 3. Immune response, defined as a 4-fold or greater rise in SBA titer 4 to 6 weeks after dose 3 compared with prevaccination titer. Children and adult recipients of either meningococcal vaccine were more likely than controls to develop an immune response to the heterologous epidemic strain. After 3 doses of vaccine, 31% to 35% of children responded to the vaccine vs 5% to placebo; 37% to 60% of adults responded to vaccine vs 4% to placebo (P<.05 vs control for all). Infants, however, did not respond. In contrast, against homologous vaccine type strains, the response rate was 67% or higher among children and adults and 90% or higher among infants (P<.001 vs control for all). Subsequent SBA against 7 isogenic homologous target strains identified class 1 OMP as the immunodominant antigen. These data suggest that neither serogroup B OMP meningococcal vaccine would confer protection during a heterologous epidemic. However, epidemic strain-specific vaccines homologous for class 1 OMP are promising candidates for the control of epidemic serogroup B meningococcal disease.

Carbapenem-Resistant Strain of Klebsiella oxytoca Harboring Carbapenem-Hydrolyzing β-Lactamase KPC-2

PubMed Central

Yigit, Hesna; Queenan, Anne Marie; Rasheed, J. Kamile; Biddle, James W.; Domenech-Sanchez, Antonio; Alberti, Sebastian; Bush, Karen; Tenover, Fred C.

2003-01-01

We investigated a Klebsiella oxytoca isolate demonstrating resistance to imipenem, meropenem, extended-spectrum cephalosporins, and aztreonam. The MICs of both imipenem and meropenem were 32 μg/ml. The β-lactamase activity against imipenem and meropenem was inhibited in the presence of clavulanic acid. Isoelectric focusing studies demonstrated five β-lactamases with pIs of 8.2 (SHV-46), 6.7 (KPC-2), 6.5 (unknown), 6.4 (probable OXY-2), and 5.4 (TEM-1). The presence of the blaSHV and blaTEM genes was confirmed by specific PCR assays and DNA sequence analysis. Transformation and conjugation studies with Escherichia coli showed that the β-lactamase with a pI of 6.7, Klebsiella pneumoniae carbapenemase-2 (KPC-2), was encoded on an approximately 70-kb conjugative plasmid that also carried SHV-46, TEM-1, and the β-lactamase with a pI of 6.5. The blaKPC-2 determinant was cloned in E. coli and conferred resistance to imipenem, meropenem, extended-spectrum cephalosporins, and aztreonam. The amino acid sequence of KPC-2 showed a single amino acid difference, S174G, when compared with KPC-1, another carbapenem-hydrolyzing β-lactamase from K. pneumoniae 1534. Hydrolysis studies showed that purified KPC-2 hydrolyzed not only carbapenems but also penicillins, cephalosporins, and aztreonam. KPC-2 had the highest affinity for meropenem. The kinetic studies revealed that KPC-2 was inhibited by clavulanic acid and tazobactam. An examination of the outer membrane proteins of the parent K. oxytoca strain demonstrated that it expressed detectable levels of OmpK36 (the homolog of OmpC) and a higher-molecular-weight OmpK35 (the homolog of OmpF). Thus, carbapenem resistance in K. oxytoca 3127 is due to production of the Bush group 2f, class A, carbapenem-hydrolyzing β-lactamase KPC-2. This β-lactamase is likely located on a transposon that is part of a conjugative plasmid and thus has a very high potential for dissemination. PMID:14638498

DOE Office of Scientific and Technical Information (OSTI.GOV)

Vikraman, S; Ramu, M; Karrthick, Kp

Purpose: The purpose of this study was to validate the advent of COMPASS 3D dosimetry as a routine pre treatment verification tool with commercially available CMS Monaco and Oncentra Masterplan planning system. Methods: Twenty esophagus patients were selected for this study. All these patients underwent radical VMAT treatment in Elekta Linac and plans were generated in Monaco v5.0 with MonteCarlo(MC) dose calculation algorithm. COMPASS 3D dosimetry comprises an advanced dose calculation algorithm of collapsed cone convolution(CCC). To validate CCC algorithm in COMPASS, The DICOM RT Plans generated using Monaco MC algorithm were transferred to Oncentra Masterplan v4.3 TPS. Only finalmore » dose calculations were performed using CCC algorithm with out optimization in Masterplan planning system. It is proven that MC algorithm is an accurate algorithm and obvious that there will be a difference with MC and CCC algorithms. Hence CCC in COMPASS should be validated with other commercially available CCC algorithm. To use the CCC as pretreatment verification tool with reference to MC generated treatment plans, CCC in OMP and CCC in COMPASS were validated using dose volume based indices such as D98, D95 for target volumes and OAR doses. Results: The point doses for open beams were observed <1% with reference to Monaco MC algorithms. Comparisons of CCC(OMP) Vs CCC(COMPASS) showed a mean difference of 1.82%±1.12SD and 1.65%±0.67SD for D98 and D95 respectively for Target coverage. Maximum point dose of −2.15%±0.60SD difference was observed in target volume. The mean lung dose of −2.68%±1.67SD was noticed between OMP and COMPASS. The maximum point doses for spinal cord were −1.82%±0.287SD. Conclusion: In this study, the accuracy of CCC algorithm in COMPASS 3D dosimetry was validated by compared with CCC algorithm in OMP TPS. Dose calculation in COMPASS is feasible within < 2% in comparison with commercially available TPS algorithms.« less

Toward Affordable Systems: Portfolio Analysis and Management for Army Science and Technology Programs

DTIC Science & Technology

2009-01-01

PROGRAM ELEMENT NUMBER 6. AUTHOR (S) 5d. PROJECT NUMBER 5e. TASK NUMBER 5f. WORK UNIT NUMBER 7. PERFORMING ORGANIZATION NAME(S) AND ADDRESS(ES) Rand...Decision Authority MDAP major defense acquisition program Abbreviations xxvii MIC marginal implementation cost MOMC marginal operating and...the Milestone Decision Authority (MDA).1 1 This section is based on U.S. Army, 2003, pp. 30–33. This 2003 Army document had taken into account the

Global budgets in Maryland: early evidence on revenues, expenses, and margins in regulated and unregulated services.

PubMed

Malmmose, Margit; Mortensen, Karoline; Holm, Claus

2018-04-02

Maryland implemented one of the most aggressive payment innovations the nation has seen in several decades when it introduced global budgets in all its acute care hospitals in 2014. Prior to this, a pilot program, total patient revenue (TPR), was established for 8 rural hospitals in 2010. Using financial hospital report data from the Health Services Cost Review Commission from 2007 to 2013, we examined the hospitals' financial results including revenue, costs, and profit/loss margins to explore the impact of the adoption of the TPR pilot global budget program relative to the remaining hospitals in the state. We analyze financial results for both regulated (included in the global budget and subject to rate-setting) and unregulated services in order to capture a holistic image of the hospitals' actual revenue, cost and margin structures. Common size and difference-in-differences analyses of the data suggest that regulated profit ratios for treatment hospitals increased (from 5% in 2007 to 8% in 2013) and regulated expense-to-gross patient revenue ratios decreased (75% in 2007 and 68% in 2013) relative to the controls. Simultaneously, the profit margins for treatment hospitals' unregulated services decreased (- 12% in 2007 and - 17% in 2013), which reduced the overall margin significantly. This analysis therefore indicates cost shifting and less profit gain from the program than identified by solely focusing on the regulated margins.

New approaches to merging multi-sensor satellite measurements of volcanic SO2 emissions

NASA Astrophysics Data System (ADS)

Carn, S. A.; Telling, J. W.; Krotkov, N. A.

2015-12-01

As part of the NASA MEaSUREs program, we are developing a unique long-term database of volcanic sulfur dioxide (SO2) emissions for use by the scientific community, using observations from multiple satellite instruments collected since 1978. Challenges to creating such a database include assessing data continuity between multiple satellite missions and SO2 retrieval algorithms and estimating measurement uncertainties. Here, we describe the approaches that we are using to merge multi-decadal SO2 measurements from the ultraviolet (UV) Total Ozone Mapping Spectrometer (TOMS), Ozone Monitoring Instrument (OMI) and Ozone Monitoring and Profiler Suite (OMPS) sensors. A particular challenge has involved accounting for the OMI row anomaly (ORA), a data gap in OMI measurements since 2008 that partially or wholly obscures some volcanic eruption clouds, whilst still profiting from the high OMI spatial resolution and data quality, and prior OMI SO2 validation. We present a new method to substitute missing SO2 information in the ORA with near-coincident SO2 data from OMPS, providing improved estimates of eruptive volcanic SO2 emissions. The technique can also be used to assess consistency between different satellite instruments and SO2 retrieval algorithms, investigate the impact of variable sensor spatial resolution, and estimate measurement uncertainties. It is particularly effective for larger eruptions producing extensive SO2 clouds where the ORA obscures the volcanic plume in multiple contiguous orbits. Application of the technique is demonstrated using recent volcanic eruptions including the 2015 eruption of Calbuco, Chile. We also provide an update on the status of the multi-satellite long-term volcanic SO2 database (MSVOLSO2L4).

Enabling the Continuous EOS-SNPP Satellite Data Record thru EOSDIS Services

NASA Astrophysics Data System (ADS)

Hall, A.; Behnke, J.; Ho, E. L.

2015-12-01

Following Suomi National Polar-Orbiting Partnership (SNPP) launch of October 2011, the role of the NASA Science Data Segment (SDS) focused primarily on evaluation of the sensor data records (SDRs) and environmental data records (EDRs) produced by the Joint Polar Satellite System (JPSS), a National Oceanic and Atmosphere Administration (NOAA) Program as to their suitability for Earth system science. The evaluation has been completed for Visible Infrared Imager Radiometer Suite (VIIRS), Advanced Technology Microwave Sounder (ATMS), Cross-track Infrared Sounder (CrIS), and Ozone Mapper/Profiler Suite (OMPS) Nadir instruments. Since launch, the SDS has also been processing, archiving and distributing data from the Clouds and the Earth's Radiant Energy System (CERES) and Ozone Mapper/Profiler Suite (OMPS) Limb instruments and this work is planned to continue through the life of the mission. As NASA transitions to the production of standard, Earth Observing System (EOS)-like science products for all instruments aboard Suomi NPP, the Suomi NPP Science Team (ST) will need data processing and production facilities to produce the new science products they develop. The five Science Investigator-led Processing Systems (SIPS): Land, Ocean. Atmosphere, Ozone, and Sounder will produce the NASA SNPP standard Level 1, Level 2, and global Level 3 products and provide the products to the NASA's Distributed Active Archive Centers (DAACs) for distribution to the user community. The SIPS will ingest EOS compatible Level 0 data from EOS Data Operations System (EDOS) for their data processing. A key feature is the use of Earth Observing System Data and Information System (EOSDIS) services for the continuous EOS-SNPP satellite data record. This allows users to use the same tools and interfaces on SNPP as they would on the entire NASA Earth Science data collection in EOSDIS.

Color perception and ATC job performance.

DOT National Transportation Integrated Search

1983-07-01

Current OMP policy and guidance requires demonstrated job-relatedness and reasonable accommodation in the application of physical qualifications. The OPM has accomplished an analysis of the Air Traffic Control Specialist (ATCS) series and recommended...

Investigations on Rickettsia in Ticks at the Sino-Russian and Sino-Mongolian Borders, China.

PubMed

Liu, Lijuan; Chen, Qian; Yang, Yu; Wang, Jiancheng; Cao, Xiaomei; Zhang, Sheng; Li, Hong; Hou, Yong; Wang, Fuxiang; Xu, Baoliang

2015-12-01

To describe the prevalence of Rickettsia in ticks at the Sino-Russian and Sino-Mongolian borders, a total of 292 ticks were collected and tested by conventional PCR assays. The prevalence of Rickettsia was 53.4%, and phylogenetic analysis showed that they belonged to R. raoultii species after alignment for the ompA, ompB, and gltA genes, respectively. Coxiella burnetii DNA was detected for 14%, and no Ehrlichia, Borrelia burgdorferi, and Babesia species were found. Co-infection of two pathogens was 9.9%, and no co-infection with three or more pathogens was found. This study suggested Rickettsia was the most common pathogen in the ticks and co-infection was found. The findings might be helpful to provide advice on the prevention and control of tick-borne disease potential for tourists and residents.

Immunohistochemical studies for the neuronal elements in the vomeronasal organ of the one-humped camel

PubMed Central

IBRAHIM, Dalia; ABDEL-MAKSOUD, Fatma; TANIGUCHI, Kazumi; YAMAMOTO, Yoshio; TANIGUCHI, Kazuyuki; NAKAMUTA, Nobuaki

2014-01-01

The neuronal elements of the vomeronasal organ (VNO) of camel were investigated immunohistochemically. PGP 9.5 labeled the receptor cells in the vomeronasal sensory epithelium, but not the supporting or basal cells. OMP stained some receptor cells, but no immunoreactive signals for OMP were detected in the non-sensory epithelium. PLCβ2 labeled scattered cells in the sensory epithelium and a larger number of cells in the non-sensory epithelium. Double labeling immunohistochemistry revealed that the PLCβ2-positive cells were surrounded by substance P-positive nerve fibers. Collectively, these data suggest that the camel VNO bears, in addition to the mature vomeronasal receptor cells, trigeminally-innervated solitary chemosensory cells which are expected to play a substantial role in the control of stimulus access to the VNO. PMID:25319516

Exposure to Glycolytic Carbon Sources Reveals a Novel Layer of Regulation for the MalT Regulon

PubMed Central

Reimann, Sylvia A.; Wolfe, Alan J.

2011-01-01

Bacteria adapt to changing environments by means of tightly coordinated regulatory circuits. The use of synthetic lethality, a genetic phenomenon in which the combination of two nonlethal mutations causes cell death, facilitates identification and study of such circuitry. In this study, we show that the E. coli ompR malT con double mutant exhibits a synthetic lethal phenotype that is environmentally conditional. MalTcon, the constitutively active form of the maltose system regulator MalT, causes elevated expression of the outer membrane porin LamB, which leads to death in the absence of the osmoregulator OmpR. However, the presence and metabolism of glycolytic carbon sources, such as sorbitol, promotes viability and unveils a novel layer of regulation within the complex circuitry that controls maltose transport and metabolism. PMID:21912549

Exposure to Glycolytic Carbon Sources Reveals a Novel Layer of Regulation for the MalT Regulon.

PubMed

Reimann, Sylvia A; Wolfe, Alan J

2011-01-01

Bacteria adapt to changing environments by means of tightly coordinated regulatory circuits. The use of synthetic lethality, a genetic phenomenon in which the combination of two nonlethal mutations causes cell death, facilitates identification and study of such circuitry. In this study, we show that the E. coli ompR malT(con) double mutant exhibits a synthetic lethal phenotype that is environmentally conditional. MalT(con), the constitutively active form of the maltose system regulator MalT, causes elevated expression of the outer membrane porin LamB, which leads to death in the absence of the osmoregulator OmpR. However, the presence and metabolism of glycolytic carbon sources, such as sorbitol, promotes viability and unveils a novel layer of regulation within the complex circuitry that controls maltose transport and metabolism.

Immunological study of an attenuated Salmonella Typhimurium expressing ApxIA, ApxIIA, ApxIIIA and OmpA of Actinobacillus pleuropneumoniae in a mouse model.

PubMed

Hur, Jin; Eo, Seong Kug; Park, Sang-Youel; Choi, Yoonyoung; Lee, John Hwa

2016-01-01

Salmonella Typhimurium strain expressing the Actinobacillus pleuropneumoniae antigens, ApxIA, ApxIIA, ApxIIIA and OmpA, was previously constructed as a vaccine candidate for porcine pleuropneumonia. This strain was a live attenuated (∆lon∆cpxR∆asd)Salmonella as a delivery host and contained a vector containing asd. An immunological study of lymphocyte proliferation, T-lymphocyte subsets and cytokines in the splenocytes of a mouse model was carried out after stimulation with the candidate Salmonella Typhimurium by intranasal inoculation. The splenic lymphocyte proliferation and the levels of IL-4, IL-6 and IL-12 of the inoculated mice were significantly increased, and the T- and B-cell populations were also elevated. Collectively, the candidate may efficiently induce the Th1- and Th2-type immune responses.

A novel electrochemical sensor based on zirconia/ordered macroporous polyaniline for ultrasensitive detection of pesticides.

PubMed

Wang, Yonglan; Jin, Jun; Yuan, Caixia; Zhang, Fan; Ma, Linlin; Qin, Dongdong; Shan, Duoliang; Lu, Xiaoquan

2015-01-21

A simple and mild strategy was proposed to develop a novel electrochemical sensor based on zirconia/ordered macroporous polyaniline (ZrO2/OMP) and further used for the detection of methyl parathion (MP), one of the organophosphate pesticides (OPPs). Due to the strong affinity of phosphate groups with ZrO2 and the advantages of OMP such as high catalytic activity and good conductivity, the developed sensor showed a limit of detection as low as 2.28 × 10(-10) mol L(-1) (S/N = 3) by square-wave voltammograms, and good selectivity, acceptable reproducibility and stability. Most importantly, this novel sensor was successfully applied to detect MP in real samples of apple and cabbage. It is expected that this method has potential applications in electrochemical sensing platforms with simple, sensitive, selective and fast analysis.

From LIMS to OMPS-LP: Limb Ozone Observations for Future Reanalyses

NASA Technical Reports Server (NTRS)

Wargan, K.; Kramarova, N.; Remsberg, E.; Coy, L.; Harvey, L.; Livesey, N.; Pawson, S.

2017-01-01

High vertical resolution and accuracy of ozone data from satellite-borne limb sounders has made them an invaluable tool in scientific studies of the middle and upper atmosphere. However, it was not until recently that these measurements were successfully incorporated in atmospheric reanalyses: of the major multidecadal reanalyses only ECMWF's (European Centre for Medium-Range Weather Forecasts') ERA (ECMWF Re-Analysis)-Interim/ERA5 and NASA's MERRA-2 (Modern-Era Retrospective Analysis for Research and Applications-2) use limb ozone data. Validation and comparison studies have demonstrated that the addition of observations from the Microwave Limb Sounder (MLS) on EOS (Earth Observing System) Aura greatly improved the quality of ozone fields in MERRA-2 making these assimilated data sets useful for scientific research. In this presentation, we will show the results of test experiments assimilating retrieved ozone from the Limb Infrared Monitor of the Stratosphere (LIMS, 1978/1979) and Ozone Mapping Profiler Suite Limb Profiler (OMPS-LP, 2012 to present). Our approach builds on the established assimilation methodology used for MLS in MERRA-2 and, in the case of OMPS-LP, extends the excellent record of MLS ozone assimilation into the post-EOS era in Earth observations. We will show case studies, discuss comparisons of the new experiments with MERRA-2, strategies for bias correction and the potential for combined assimilation of multiple limb ozone data types in future reanalyses for studies of multidecadal stratospheric ozone changes including trends.

Improvement of n-butanol tolerance in Escherichia coli by membrane-targeted tilapia metallothionein

PubMed Central

2013-01-01

Background Though n-butanol has been proposed as a potential transportation biofuel, its toxicity often causes oxidative stress in the host microorganism and is considered one of the bottlenecks preventing its efficient mass production. Results To relieve the oxidative stress in the host cell, metallothioneins (MTs), which are known as scavengers for reactive oxygen species (ROS), were engineered in E. coli hosts for both cytosolic and outer-membrane-targeted (osmoregulatory membrane protein OmpC fused) expression. Metallothioneins from human (HMT), mouse (MMT), and tilapia fish (TMT) were tested. The host strain expressing membrane-targeted TMT showed the greatest ability to reduce oxidative stresses induced by n-butanol, ethanol, furfural, hydroxymethylfurfural, and nickel. The same strain also allowed for an increased growth rate of recombinant E. coli under n-butanol stress. Further experiments indicated that the TMT-fused OmpC protein could not only function in ROS scavenging but also regulate either glycine betaine (GB) or glucose uptake via osmosis, and the dual functional fusion protein could contribute in an enhancement of the host microorganism’s growth rate. Conclusions The abilities of scavenging intracellular or extracellular ROS by these engineering E. coli were examined, and TMT show the best ability among three MTs. Additionally, the membrane-targeted fusion protein, OmpC-TMT, improved host tolerance up to 1.5% n-butanol above that of TMT which is only 1%. These results presented indicate potential novel approaches for engineering stress tolerant microorganism strains. PMID:24020941

Improvement of n-butanol tolerance in Escherichia coli by membrane-targeted tilapia metallothionein.

PubMed

Chin, Wei-Chih; Lin, Kuo-Hsing; Chang, Jui-Jen; Huang, Chieh-Chen

2013-09-11

Though n-butanol has been proposed as a potential transportation biofuel, its toxicity often causes oxidative stress in the host microorganism and is considered one of the bottlenecks preventing its efficient mass production. To relieve the oxidative stress in the host cell, metallothioneins (MTs), which are known as scavengers for reactive oxygen species (ROS), were engineered in E. coli hosts for both cytosolic and outer-membrane-targeted (osmoregulatory membrane protein OmpC fused) expression. Metallothioneins from human (HMT), mouse (MMT), and tilapia fish (TMT) were tested. The host strain expressing membrane-targeted TMT showed the greatest ability to reduce oxidative stresses induced by n-butanol, ethanol, furfural, hydroxymethylfurfural, and nickel. The same strain also allowed for an increased growth rate of recombinant E. coli under n-butanol stress. Further experiments indicated that the TMT-fused OmpC protein could not only function in ROS scavenging but also regulate either glycine betaine (GB) or glucose uptake via osmosis, and the dual functional fusion protein could contribute in an enhancement of the host microorganism's growth rate. The abilities of scavenging intracellular or extracellular ROS by these engineering E. coli were examined, and TMT show the best ability among three MTs. Additionally, the membrane-targeted fusion protein, OmpC-TMT, improved host tolerance up to 1.5% n-butanol above that of TMT which is only 1%. These results presented indicate potential novel approaches for engineering stress tolerant microorganism strains.

Product deuterium isotope effects for orotidine 5'-monophosphate decarboxylase: effect of changing substrate and enzyme structure on the partitioning of the vinyl carbanion reaction intermediate.

PubMed

Toth, Krisztina; Amyes, Tina L; Wood, Bryant M; Chan, Kui; Gerlt, John A; Richard, John P

2010-05-26

A product deuterium isotope effect (PIE) of 1.0 was determined as the ratio of the yields of [6-(1)H]-uridine 5'-monophosphate (50%) and [6-(2)H]-uridine 5'-monophosphate (50%) from the decarboxylation of orotidine 5'-monophosphate (OMP) in 50/50 (v/v) HOH/DOD catalyzed by orotidine 5'-monophosphate decarboxylase (OMPDC) from Saccharomyces cerevisiae, Methanothermobacter thermautotrophicus, and Escherichia coli. This unitary PIE eliminates a proposed mechanism for enzyme-catalyzed decarboxylation in which proton transfer from Lys-93 to C-6 of OMP provides electrophilic push to the loss of CO(2) in a concerted reaction. We propose that the complete lack of selectivity for the reaction of solvent H and D, which is implied by the value of PIE = 1.0, is enforced by restricted C-N bond rotation of the -CH(2)-NL(3)(+) group of the side chain of Lys-93. A smaller PIE of 0.93 was determined as the ratio of the product yields for OMPDC-catalyzed decarboxylation of 5-fluoroorotidine 5'-monophosphate (5-FOMP) in 50/50 (v/v) HOH/DOD. Mutations on the following important active-site residues of OMPDC from S. cerevisiae have no effect on the PIE on OMPDC-catalyzed decarboxylation of OMP or decarboxylation of 5-FOMP: R235A, Y217A, Q215A, S124A, and S154A/Q215A.

Product Deuterium Isotope Effects for Orotidine 5'-Monophosphate Decarboxylase: Effect of Changing Substrate and Enzyme Structure on the Partitioning of the Vinyl Carbanion Reaction Intermediate

PubMed Central

Toth, Krisztina; Amyes, Tina L.; Wood, Bryant M.; Chan, Kui; Gerlt, John A.

2010-01-01

A product deuterium isotope effect (PIE) of 1.0 was determined as the ratio of the yields of [6-1H]-uridine 5'-monophosphate (50%) and [6-2H]-uridine 5'-monophosphate (50%) from the decarboxylation of orotidine 5'-monophosphate (OMP) in 50/50 (v/v) HOH/DOD catalyzed by orotidine 5'-monophosphate decarboxylase (OMPDC) from S. cerevisiae, M. thermautotrophicus and E. coli. This unitary PIE eliminates a proposed mechanism for enzyme-catalyzed decarboxylation in which proton transfer from Lys-93 to C-6 of OMP provides electrophilic push to the loss of CO2 in a concerted reaction. We propose that the complete lack of selectivity for the reaction of solvent H and D, that is implied by the value of PIE = 1.0, is enforced by restricted C-N bond rotation of the -CH2-NL3+ group of the side-chain of Lys-93. A smaller PIE of 0.93 was determined as the ratio of the product yields for OMPDC-catalyzed decarboxylation of 5-fluoroorotidine 5'-monophosphate (5-FOMP) in 50/50 (v/v) HOH/DOD. The following mutations of important active site residues of OMPDC from S. cerevisiae have no effect on the PIE on OMPDC-catalyzed decarboxylation of OMP or decarboxylation of 5-FOMP: R235A, Y217A, Q215A, S124A and S154A/Q215A. PMID:20441167

Correlation between direction and severity of temporomandibular joint disc displacement and reduction ability during mouth opening.

PubMed

Litko, M; Berger, M; Szkutnik, J; Różyło-Kalinowska, I

2017-12-01

The most common temporomandibular joint (TMJ) internal derangement is an abnormal relationship of the disc with respect to the mandibular condyle, articular eminence and glenoid fossa-disc displacement. The aim of our study was to analyse the correlation between partial/complete disc displacement in the intercuspal position (IP) and its reduction in the open-mouth position (OMP) in both oblique sagittal and coronal planes on magnetic resonance imaging (MRI) in patients with temporomandibular disorders. Multisection MRI analysis of 382 TMJs was conducted in 191 patients with disc displacement according to the RDC/TMD criteria (148 women, 43 men; aged 14-60 years). The disc position was evaluated on all oblique sagittal and coronal images in the IP and the OMP. Univariate logistic regression analysis showed that the severity of disc displacement in the sagittal plane is a statistically significant predictor of reduction ability during mouth opening (B = 3.118; P < .001). Moreover, the severity of disc displacement in both planes is also a significant predictor of disc reduction in OMP (B = 2.200; P < .05). In conclusion, reduction ability during mouth opening is associated with the severity of disc displacement in IP, in both sagittal and coronal planes. Multisection analysis of all MR images allows distinguishing the correct disc position from disc displacement and can improve the ability to distinguish between various stages of TMJ internal derangement. © 2017 John Wiley & Sons Ltd.

The Escherichia coli O157:H7 cattle immuno-proteome includes outer membrane protein A (OmpA), a modulator of adherence to bovine recto-anal junction squamous epithelial (RSE) cells

PubMed Central

Kudva, Indira T.; Krastins, Bryan; Torres, Alfredo G.; Griffin, Robert W.; Sheng, Haiqing; Sarracino, David A.; Hovde, Carolyn J.; Calderwood, Stephen B.; John, Manohar

2015-01-01

SUMMARY Building on previous studies, we defined the repertoire of proteins comprising the immuno-proteome of E. coli O157:H7 (O157) cultured in DMEM supplemented with norepinephrine (NE; O157 immuno-proteome), a β-adrenergic hormone that regulates E. coli O157 gene expression in the gastrointestinal tract, using a variation of a novel proteomics-based platform proteome mining tool for antigen discovery, called Proteomics-based Expression Library Screening (PELS; Kudva et al., 2006). The E. coli O157 immuno-proteome (O157-IP) comprised 91 proteins, and included those identified previously using PELS, and also proteins comprising DMEM- and bovine rumen fluid- proteomes. Outer membrane protein A (OmpA), a common component of the above proteomes, and reportedly a contributor to E. coli O157 adherence to cultured Hep-2 epithelial cells, was interestingly found to be a modulator rather than a contributor to E. coli O157 adherence to bovine recto-anal junction squamous epithelial (RSE) cells. Our results point to a role for yet to be identified members of the O157-IP in E. coli O157 adherence to RSE-cells, and additionally implicate a possible role for the OmpA regulator, TdcA, in the expression of such adhesins. Our observations have implications for development of efficacious vaccines for preventing E. coli O157 colonization of the bovine gastrointestinal tract. PMID:25643951

Evaluation of immunogenicity and protective efficacy of recombinant outer membrane proteins of Haemophilus parasuis serovar 5 in a murine model

PubMed Central

Li, Miao; Cai, Ru-Jian; Song, Shuai; Jiang, Zhi-Yong; Li, Yan; Gou, Hong-Chao; Chu, Pin-Pin; Li, Chun-Ling; Qiu, Hua-Ji

2017-01-01

Glässer’s disease is an economically important infectious disease of pigs caused by Haemophilus parasuis. Few vaccines are currently available that could provide effective cross-protection against various serovars of H. parasuis. In this study, five OMPs (OppA, TolC, HxuC, LppC, and HAPS_0926) identified by bioinformatic approaches, were cloned and expressed as recombinant proteins. Antigenicity of the purified proteins was verified through Western blotting, and primary screening for protective potential was evaluated in vivo. Recombinant TolC (rTolC), rLppC, and rHAPS_0926 proteins showing marked protection of mice against H. parasuis infection, and were further evaluated individually or in combination. Mice treated with these three OMPs produced humoral and host cell-mediated responses, with a significant rise in antigen-specific IgG titer and lymphoproliferative response in contrast with the mock-immunized group. Significant increases were noted in CD4+, CD8+ T cells, and three cytokines (IL-2, IL-4, and IFN-γ) in vaccinated animals. The antisera against candidate antigens could efficiently impede bacterial survival in whole blood bactericidal assay against H. parasuis infection. The multi-protein vaccine induced more pronounced immune responses and offered better protection than individual vaccines. Our findings indicate that these three OMPs are promising antigens for the development of multi-component subunit vaccines against Glässer's disease. PMID:28448603

Teaching the One-minute Preceptor

PubMed Central

Furney, Scott L; Orsini, Alex N; Orsetti, Kym E; Stern, David T; Gruppen, Larry D; Irby, David M

2001-01-01

OBJECTIVE The One-Minute Preceptor (OMP) model of faculty development is used widely to improve teaching, but its effect on teaching behavior has not been assessed. We aim to evaluate the effect of this intervention on residents' teaching skills. DESIGN Randomized controlled trial. SETTING Inpatient teaching services at both a tertiary care hospital and a Veterans Administration Medical Center affiliated with a University Medical Center. PARTICIPANTS Participants included 57 second- and third-year internal medicine residents that were randomized to the intervention group (n = 28) or to the control group (n = 29). INTERVENTION The intervention was a 1-hour session incorporating lecture, group discussion, and role-play. MEASUREMENTS AND MAIN RESULTS Primary outcome measures were resident self-report and learner ratings of resident performance of the OMP teaching behaviors. Residents assigned to the intervention group reported statistically significant changes in all behaviors (P < .05). Eighty-seven percent of residents rated the intervention as “useful or very useful” on a 1–5 point scale with a mean of 4.28. Student ratings of teacher performance showed improvements in all skills except “Teaching General Rules.” Learners of the residents in the intervention group reported increased motivation to do outside reading when compared to learners of the control residents. Ratings of overall teaching effectiveness were not significantly different between the 2 groups. CONCLUSIONS The OMP model is a brief and easy-to-administer intervention that provides modest improvements in residents' teaching skills. PMID:11556943

A Two-Stage Reconstruction Processor for Human Detection in Compressive Sensing CMOS Radar

PubMed Central

Tsao, Kuei-Chi; Lee, Ling; Chu, Ta-Shun

2018-01-01

Complementary metal-oxide-semiconductor (CMOS) radar has recently gained much research attraction because small and low-power CMOS devices are very suitable for deploying sensing nodes in a low-power wireless sensing system. This study focuses on the signal processing of a wireless CMOS impulse radar system that can detect humans and objects in the home-care internet-of-things sensing system. The challenges of low-power CMOS radar systems are the weakness of human signals and the high computational complexity of the target detection algorithm. The compressive sensing-based detection algorithm can relax the computational costs by avoiding the utilization of matched filters and reducing the analog-to-digital converter bandwidth requirement. The orthogonal matching pursuit (OMP) is one of the popular signal reconstruction algorithms for compressive sensing radar; however, the complexity is still very high because the high resolution of human respiration leads to high-dimension signal reconstruction. Thus, this paper proposes a two-stage reconstruction algorithm for compressive sensing radar. The proposed algorithm not only has lower complexity than the OMP algorithm by 75% but also achieves better positioning performance than the OMP algorithm especially in noisy environments. This study also designed and implemented the algorithm by using Vertex-7 FPGA chip (Xilinx, San Jose, CA, USA). The proposed reconstruction processor can support the 256×13 real-time radar image display with a throughput of 28.2 frames per second. PMID:29621170

Prevalence and characterization of Chlamydia DNA in zoo animals in Japan.

PubMed

Kabeya, Hidenori; Sato, Shingo; Maruyama, Soichi

2015-09-01

Because many people visit zoos, prevention of zoonoses is important from the standpoint of public health. This study examined the prevalence of Chlamydia among zoo animals in Japan by PCR and characterized these bacteria by performing phylogenetic analyses of the sequences of the variable domain (VD) 2 and VD4 regions of the ompA gene, which encodes the Chlamydia major outer membrane protein. Fecal samples were collected from 1150 zoo animals in five zoos and examined for Chlamydia DNA. Chlamydia psittaci DNA was found in 3.9% of mammals, 7.2% of birds and 8.1% of reptiles. The prevalence of Chlamydia pneumoniae DNA was significantly higher in reptiles (5.8%) than in mammals (0.3%) and birds (0.3%). Phylogenetic analysis of the ompA VD2 region from 18 samples showed that nine were in three different clusters containing C. psittaci strains, six were in a cluster containing C. pneumoniae strains and three each formed a distinct branch. Furthermore, phylogenetic analysis of the ompA VD4 region showed that C. pneumoniae DNAs from reptiles were close to those from human patients. The C. pneumoniae DNAs from the European glass lizard, Emerald tree boa, and Panther chameleon were classified in clusters that were distinct from other strains, suggesting that these reptiles had each been infected with a specific C. pneumoniae genotype. This study showed that diverse Chlamydia strains have been prevalent among a variety of zoo animals. © 2015 The Societies and Wiley Publishing Asia Pty Ltd.

PCR-based diagnosis, molecular characterization and detection of atypical strains of avian Chlamydia psittaci in companion and wild birds.

PubMed

Madani, S A; Peighambari, S M

2013-02-01

Chlamydiosis is one of the most important infectious diseases of birds. In this study, 253 clinical samples were taken from 27 bird species belonging to seven orders. Thirty-two (12.6%) samples were positive for Chlamydia psittaci major outer membrane gene (ompA) DNA by a nested polymerase chain reaction (PCR). Twelve nested PCR-positive specimens were typed by ompA gene-based PCR-restricted fragment length polymorphism, using CTU/CTL primers and AluI restriction enzyme. Four restriction patterns were identified, including genotype A (two specimens from an African grey parrot [Psittacus erithacus] and a lorikeet [Trichoglossus haematodus]), genotype B (two specimens from a rock dove [Columbia livia] and a canary [Serinus canaria]), a third new restriction pattern (six specimens from African grey parrots), and a fourth new restriction pattern (two specimens from a ring-necked parakeet [Psittacula krameri] and an Alexandrine parakeet [Psittacula eupatria]). The third and the fourth restriction patterns are suggested to be provisional genotypes I and J, respectively. Partial sequencing of the ompA gene of seven specimens completely correlated with the results of PCR-restricted fragment length polymorphism and confirmed the presence of genotypes A and B and the two new provisional genotypes I and J. The two new genotypes have the closest identity with C. psittaci genotype F and Chlamydia abortus, respectively. From an evolutionary perspective, both new genotypes, particularly genotype J, are intermediate between the two species, C. psittaci and C. abortus.

76 FR 52937 - Ball Bearings and Parts Thereof From France, Germany, and Italy: Final Results of Antidumping...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-08-24

... programming and other errors in the margin calculations. Therefore, the final results are different from the... certain companies. We have corrected programming and other errors in the margins we included in the... Technologies GmbH; Italy--Schaeffler Italia S.r.l./WPB Water Pump Bearing GmbH & Co. KG/The Schaeffler Group...

The Marginal Effects of High School Visits: A Step toward an Empirically Driven Marketing Program. AIR 2002 Forum Paper.

ERIC Educational Resources Information Center

Aguilar, Lore; Gillespie, Donald A.

Admissions officers devote substantial resources to high school visits and other recruiting events. Economic theory indicates that the best measure of effectiveness is marginal productivity. Subjects in this study were applicants to a large, Catholic, urban university and all students who participated in recruiting programs in fall 2002.…

The Single Sex Debate for Girls in Science: a Comparison Between Two Informal Science Programs on Middle School Students' STEM Identity Formation

NASA Astrophysics Data System (ADS)

Hughes, Roxanne M.; Nzekwe, Brandon; Molyneaux, Kristen J.

2013-10-01

Currently, there are policy debates regarding the efficacy and legality of single sex formal and informal education programs. This issue is particularly poignant in science education due to the historical marginalization of women in these fields. This marginalization has resulted in women being positioned as a stigmatized group within many science, technology, engineering, and mathematics (STEM) related fields. Research points to adolescence as the age where this sense of marginalization begins to develop. As a result, policy responses have utilized various frameworks such as: increased access for women, changing pedagogy to address women's learning styles, changing the language and culture of science to prevent marginalization of stigmatized groups, and finally exploring the role that individual identity plays in the marginalization of women. This study adds to the policy debate as it applies to single sex education by comparing middle school participants' STEM identity formation during two informal science learning environments (an all girls' STEM camp and a co-educational STEM camp). Additionally, this study focuses on the influence of camp activities within two informal science education programs: particularly the provision of role models and authentic STEM research activities, as means to improve STEM identity and make these fields relevant to the lives of middle school students. The results indicate that both camps improved girls' STEM identities. These findings suggest that the single sex environment is not as important to STEM identity as the pedagogy used within the program.

DRI Technical Program: Emerging Dynamics of the Marginal Ice Zone Ice, Ocean and Atmosphere Interactions in the Arctic Marginal Ice Zone. Year 3 Annual Report

DTIC Science & Technology

2014-09-30

Institution The Scottish Association for Marine Science tmaksym@whoi.edu Phil.Hwang@sams.ac.uk LONG-TERM GOALS This DRI TECHNICAL PROGRAM (Emerging...jpw28@bas.ac.uk tmaksym@whoi.edu Co-PRINCIPAL INVESTIGATOR: Byongjun (Phil) Hwang The Scottish Association for Marine Science Phil.Hwang@sams.ac.uk 2

A Collective Pursuit of Learning the Possibility to Be: The CAMP Experience Assisting Situationally Marginalized Mexican American Students to a Successful Student Identity

ERIC Educational Resources Information Center

Reyes, Reynaldo, III

2007-01-01

Many students of Mexican descent must learn how to be successful students. This study describes 5 students of Mexican descent from situationally marginalized lives who were a part of a support and retention scholarship program (College Assistance Migrant Program--CAMP). These case studies document how they perceived their learning and how they…

The Single Sex Debate for Girls in Science: A Comparison between Two Informal Science Programs on Middle School Students' STEM Identity Formation

ERIC Educational Resources Information Center

Hughes, Roxanne M.; Nzekwe, Brandon; Molyneaux, Kristen J.

2013-01-01

Currently, there are policy debates regarding the efficacy and legality of single sex formal and informal education programs. This issue is particularly poignant in science education due to the historical marginalization of women in these fields. This marginalization has resulted in women being positioned as a stigmatized group within many…

The Development of Dynamic Human Reliability Analysis Simulations for Inclusion in Risk Informed Safety Margin Characterization Frameworks

DOE Office of Scientific and Technical Information (OSTI.GOV)

Jeffrey C. Joe; Diego Mandelli; Ronald L. Boring

2015-07-01

The United States Department of Energy is sponsoring the Light Water Reactor Sustainability program, which has the overall objective of supporting the near-term and the extended operation of commercial nuclear power plants. One key research and development (R&D) area in this program is the Risk-Informed Safety Margin Characterization pathway, which combines probabilistic risk simulation with thermohydraulic simulation codes to define and manage safety margins. The R&D efforts to date, however, have not included robust simulations of human operators, and how the reliability of human performance or lack thereof (i.e., human errors) can affect risk-margins and plant performance. This paper describesmore » current and planned research efforts to address the absence of robust human reliability simulations and thereby increase the fidelity of simulated accident scenarios.« less

Power, discourse, and resistance: Poststructuralist influences in nursing.

PubMed

Holmes, Dave; Gagnon, Marilou

2018-01-01

Based on our respective research programs (psychiatry, forensic psychiatry, public health, HIV/AIDS, harm reduction) this article aims to use purposely non-conventional means to present the substantial contribution of poststructuralist perspectives to knowledge development in nursing science in general and in our current research in particular. More specifically, we call on the work of Michel Foucault and Deleuze & Guattari to politicize nursing science using examples from our empirical research programs with marginal and often highly marginalized populations. We discuss the concepts of power, discourse, and resistance to illustrate the essential contribution of poststructuralism to marginal, even "nomadic", nursing research. © 2017 John Wiley & Sons Ltd.

Comparison of SO2 and NO2 observations from OMI and OMPS from 2012 to 2016

NASA Astrophysics Data System (ADS)

Wang, Y.; Wang, J.; Xu, X.; Yang, K.

2017-12-01

Both Sulfur dioxide (SO2) and nitrogen dioxide (NO2) are precursors of PM2.5 which has significant impacts on human health. We compare observations from Ozone Monitoring Instrument (OMI) which has data gap due to row anomaly and Ozone Mapping Profiler Suite (OMPS) that is currently the only operational UV satellite sensor providing contiguous daily global coverage. In this study, we examine changes of SO2 and NO2 in several polluted regions and see both upward trends and downward trends in different areas but trends observed by the two sensors are consistent in general. Some of these upward and downward trends are associated with economic development and implementation of emission control policy. In addition, we analyzed probability distribution function of SO2 and NO2 from the two sensors and how row anomaly effect the intercomparison.

Affinity purification of bacterial outer membrane vesicles (OMVs) utilizing a His-tag mutant.

PubMed

Alves, Nathan J; Turner, Kendrick B; DiVito, Kyle A; Daniele, Michael A; Walper, Scott A

To facilitate the rapid purification of bacterial outer membrane vesicles (OMVs), we developed two plasmid constructs that utilize a truncated, transmembrane protein to present an exterior histidine repeat sequence. We chose OmpA, a highly abundant porin protein, as the protein scaffold and utilized the lac promoter to allow for inducible control of the epitope-presenting construct. OMVs containing mutant OmpA-His6 were purified directly from Escherichia coli culture media on an immobilized metal affinity chromatography (IMAC) Ni-NTA resin. This enabling technology can be combined with other molecular tools directed at OMV packaging to facilitate the separation of modified/cargo-loaded OMV from their wt counterparts. In addition to numerous applications in the pharmaceutical and environmental remediation industries, this technology can be utilized to enhance basic research capabilities in the area of elucidating endogenous OMV function. Published by Elsevier Masson SAS.

Enhancements to NASA's Land Atmosphere Near Real-Time Capability for EOS (LANCE)

NASA Technical Reports Server (NTRS)

Davies, Diane; Michael, Karen; Schmaltz, Jeffrey; Boller, Ryan A.; Masuoka, Ed; Ye, Gang; Roman, Miguel; Vermote, Eric; Harrison, Sherry; Rinsland, Pamela;

Validated spectrophotometric methods for simultaneous determination of Omeprazole, Tinidazole and Doxycycline in their ternary mixture

NASA Astrophysics Data System (ADS)

Lotfy, Hayam M.; Hegazy, Maha A.; Mowaka, Shereen; Mohamed, Ekram Hany

2016-01-01

A comparative study of smart spectrophotometric techniques for the simultaneous determination of Omeprazole (OMP), Tinidazole (TIN) and Doxycycline (DOX) without prior separation steps is developed. These techniques consist of several consecutive steps utilizing zero/or ratio/or derivative spectra. The proposed techniques adopt nine simple different methods, namely direct spectrophotometry, dual wavelength, first derivative-zero crossing, amplitude factor, spectrum subtraction, ratio subtraction, derivative ratio-zero crossing, constant center, and successive derivative ratio method. The calibration graphs are linear over the concentration range of 1-20 μg/mL, 5-40 μg/mL and 2-30 μg/mL for OMP, TIN and DOX, respectively. These methods are tested by analyzing synthetic mixtures of the above drugs and successfully applied to commercial pharmaceutical preparation. The methods that are validated according to the ICH guidelines, accuracy, precision, and repeatability, were found to be within the acceptable limits.

Revealed preferences towards the appraisal of orphan drugs in Poland - multi criteria decision analysis.

PubMed

Kolasa, Katarzyna; Zwolinski, Krzysztof Miroslaw; Zah, Vladimir; Kaló, Zoltán; Lewandowski, Tadeusz

2018-04-27

A Multi Criteria Decision Analysis (MCDA) technique was adopted to reveal the preferences of the Appraisal Body of the Polish HTA agency towards orphan drugs (OMPs). There were 34 positive and 23 negative HTA recommendations out of 54 distinctive drug-indication pairs. The MCDA matrix consisted of 13 criteria, seven of which made the most impact on the HTA process. Appraisal of clinical evidence, cost of therapy, and safety considerations were the main contributors to the HTA guidance, whilst advancement of technology and manufacturing costs made the least impact. MCDA can be regarded as a valuable tool for revealing decision makers' preferences in the healthcare sector. Given that only roughly half of all criteria included in the MCDA matrix were deemed to make an impact on the HTA process, there is certainly some room for improvement with respect to the adaptation of a new approach towards the value assessment of OMPs in Poland.

Study of orotidine 5'-monophosphate decarboxylase in complex with the top three OMP, BMP, and PMP ligands by molecular dynamics simulation.

PubMed

Jamshidi, Shirin; Jalili, Seifollah; Rafii-Tabar, Hashem

2015-01-01

Catalytic mechanism of orotidine 5'-monophosphate decarboxylase (OMPDC), one of the nature most proficient enzymes which provides large rate enhancement, has not been fully understood yet. A series of 30 ns molecular dynamics (MD) simulations were run on X-ray structure of the OMPDC from Saccharomyces cerevisiae in its free form as well as in complex with different ligands, namely 1-(5'-phospho-D-ribofuranosyl) barbituric acid (BMP), orotidine 5'-monophosphate (OMP), and 6-phosphonouridine 5'-monophosphate (PMP). The importance of this biological system is justified both by its high rate enhancement and its potential use as a target in chemotherapy. This work focuses on comparing two physicochemical states of the enzyme (protonated and deprotonated Asp91) and three ligands (substrate OMP, inhibitor, and transition state analog BMP and substrate analog PMP). Detailed analysis of the active site geometry and its interactions is properly put in context by extensive comparison with relevant experimental works. Our overall results show that in terms of hydrogen bond occupancy, electrostatic interactions, dihedral angles, active site configuration, and movement of loops, notable differences among different complexes are observed. Comparison of the results obtained from these simulations provides some detailed structural data for the complexes, the enzyme, and the ligands, as well as useful insights into the inhibition mechanism of the OMPDC enzyme. Furthermore, these simulations are applied to clarify the ambiguous mechanism of the OMPDC enzyme, and imply that the substrate destabilization and transition state stabilization contribute to the mechanism of action of the most proficient enzyme, OMPDC.

Stubborn contaminants: influence of detergents on the purity of the multidrug ABC transporter BmrA.

PubMed

Wiseman, Benjamin; Kilburg, Arnaud; Chaptal, Vincent; Reyes-Mejia, Gina Catalina; Sarwan, Jonathan; Falson, Pierre; Jault, Jean-Michel

2014-01-01

Despite the growing interest in membrane proteins, their crystallization remains a major challenge. In the course of a crystallographic study on the multidrug ATP-binding cassette transporter BmrA, mass spectral analyses on samples purified with six selected detergents revealed unexpected protein contamination visible for the most part on overloaded SDS-PAGE. A major contamination from the outer membrane protein OmpF was detected in purifications with Foscholine 12 (FC12) but not with Lauryldimethylamine-N-oxide (LDAO) or any of the maltose-based detergents. Consequently, in the FC12 purified BmrA, OmpF easily crystallized over BmrA in a new space group, and whose structure is reported here. We therefore devised an optimized protocol to eliminate OmpF during the FC12 purification of BmrA. On the other hand, an additional band visible at ∼110 kDa was detected in all samples purified with the maltose-based detergents. It contained AcrB that crystallized over BmrA despite its trace amounts. Highly pure BmrA preparations could be obtained using either a ΔacrAB E. coli strain and n-dodecyl-β-D-maltopyranoside, or a classical E. coli strain and lauryl maltose neopentyl glycol for the overexpression and purification, respectively. Overall our results urge to incorporate a proteomics-based purity analysis into quality control checks prior to commencing crystallization assays of membrane proteins that are notoriously arduous to crystallize. Moreover, the strategies developed here to selectively eliminate obstinate contaminants should be applicable to the purification of other membrane proteins overexpressed in E. coli.

Identification of unusual Chlamydia pecorum genotypes in Victorian koalas (Phascolarctos cinereus) and clinical variables associated with infection.

PubMed

Legione, Alistair R; Patterson, Jade L S; Whiteley, Pam L; Amery-Gale, Jemima; Lynch, Michael; Haynes, Leesa; Gilkerson, James R; Polkinghorne, Adam; Devlin, Joanne M; Sansom, Fiona M

2016-05-01

Chlamydia pecorum infection is a threat to the health of free-ranging koalas (Phascolarctos cinereus) in Australia. Utilizing an extensive sample archive we determined the prevalence of C. pecorum in koalas within six regions of Victoria, Australia. The ompA genotypes of the detected C. pecorum were characterized to better understand the epidemiology of this pathogen in Victorian koalas. Despite many studies in northern Australia (i.e. Queensland and New South Wales), prior Chlamydia studies in Victorian koalas are limited. We detected C. pecorum in 125/820 (15 %) urogenital swabs, but in only one ocular swab. Nucleotide sequencing of the molecular marker C. pecorum ompA revealed that the majority (90/114) of C. pecorum samples typed were genotype B. This genotype has not been reported in northern koalas. In general, Chlamydia infection in Victorian koalas is associated with milder clinical signs compared with infection in koalas in northern populations. Although disease pathogenesis is likely to be multifactorial, the high prevalence of genotype B in Victoria may suggest it is less pathogenic. All but three koalas had C. pecorum genotypes unique to southern koala populations (i.e. Victoria and South Australia). These included a novel C. pecorum ompA genotype and two genotypes associated with livestock. Regression analysis determined that significant factors for the presence of C. pecorum infection were sex and geographical location. The presence of 'wet bottom' in males and the presence of reproductive tract pathology in females were significantly associated with C. pecorum infection, suggesting variation in clinical disease manifestations between sexes.

Aspirin increases susceptibility of Helicobacter pylori to metronidazole by augmenting endocellular concentrations of antimicrobials.

PubMed

Zhang, Xiao-Ping; Wang, Wei-Hong; Tian, Yu; Gao, Wen; Li, Jiang

2009-02-28

To investigate the mechanisms of aspirin increasing the susceptibility of Helicobacter pylori (H pylori) to metronidazole. H pylori reference strain 26695 and two metronidazole-resistant isolates of H pylori were included in this study. Strains were incubated in Brucella broth with or without aspirin (1 mmol/L). The rdxA gene of H pylori was amplified by PCR and sequenced. The permeability of H pylori to antimicrobials was determined by analyzing the endocellular radioactivity of the cells after incubated with [7-(3)H]-tetracycline. The outer membrane proteins (OMPs) of H pylori 26695 were depurated and analyzed by SDS-PAGE. The expression of 5 porins (hopA, hopB, hopC, hopD and hopE) and the putative RND efflux system (hefABC) of H pylori were analyzed using real-time quantitative PCR. The mutations in rdxA gene did not change in metronidazole resistant isolates treated with aspirin. The radioactivity of H pylori increased when treated with aspirin, indicating that aspirin improved the permeability of the outer membrane of H pylori. However, the expression of two OMP bands between 55 kDa and 72 kDa altered in the presence of aspirin. The expression of the mRNA of hopA, hopB, hopC, hopD, hopE and hefA, hefB, hefC of H pylori did not change when treated with aspirin. Although aspirin increases the susceptibility of H pylori to metronidazole, it has no effect on the mutations of rdxA gene of H pylori. Aspirin increases endocellular concentrations of antimicrobials probably by altering the OMP expression.