Detection of Japanese Encephalitis Virus in Vector Mosquitoes in a Non-endemic Area, India

Japanese Encephalitis (JE) is a serious growing public health problem in India, gradually engulfing new areas. In north India, eastern districts of Uttar Pradesh had been highly endemic for JE since 1978. JE outbreak was reported to have jumped over 800 km distance from its earlier most endemic area (eastern parts of north India) to a location (Karnal) in the Haryana state of western parts of north India, during the year 1990, which never reported JE earlier.1 Later, the disease gradually spread to its adjoining districts in the state. First time, from western Uttar Pradesh, 7 cases and 4 deaths due to suspected JE were reported in 2003 from Saharanpur district. In the subsequent year, 13 deaths due to suspected viral encephalitis were reported from 26th September to 23rd October, 2004 from one village namely Khekra, from Baghpat district of Uttar Pradesh which is about 128 km west of Saharanpur. A serosurvey carried out on 24th and 25th October, 2004 among the family members of dead persons revealed hemagglutination inhibition (HI) antibodies against JE and West Nile (WN) virus. Further, an outbreak of suspected JE was reported from Saharanpur district in 2005, with 212 cases and 157 deaths. This caused matter of great concern to extend further studies in Baghpat where prevalence of flavivirus infection was already recorded in the locality. For monitoring of arbovirus activities in an area, detection of virus in human sera is cumbersome, difficult and also not desirable. Therefore, as an alternative approach, detection of virus antigen in mosquitoes by antigen capture enzyme linked immunosorbent assay (ELISA) has provided a reliable tool to comprehend the types of virus circulating in nature.2 Detecting arbovirus in mosquitos forms an important part of vector surveillance and may at times also serve as an early warning signal for outbreaks, if however linked with phonological and epidemiological studies. Vector infection and abundance were found to be good indicators of JE occurrence in surveillance studies conducted in South India.3 An attempt was thus made to detect the presence of JE virus (JEV) antigen in vector mosquitoes by ELISA method from the encephalitis-affected area of Baghpat district and to provide evidence of circulating of Japanese Encephalitis virus (JEV) in natural animal-mosquito cycle

Lungs deposition and pharmacokinetic study of submicron budesonide particles in Wistar rats intended for immediate effect in asthma

The purpose of the present investigation was to study the aerosolization, lungs deposition and pharmacokinetic study of inhalable submicron particles of budesonide in male Wistar rats. Submicron particles were prepared by antisolvent nanoprecipitation method and freeze-dried to obtain free flowing powder. The freeze-drying process yielded dry powder with desirable aerodynamic properties for inhalation therapy. An in-house model inhaler was designed to deliver medicine to lungs, optimized at dose level of 10 mg for 30 sec of fluidization. The in vitro aerosolization study demonstrates that submicron particles dissolve faster with improved aerosolization effect as compared to micronized budesonide. Both submicron and micron particles were compared for in vivo lungs dep- osition. The results showed that relatively high quantity of submicron particles reaches deep into the lungs as compared to micron particles. Most pronounced effect observed with submicron particles from pharmacokinetic parameters was the enhancement in peak plasma concentration (Cmax) by 28.85 %, and increase in area under concentration curve (AUC0–8h) by 30.33 % compared to micron sized particles. The results suggested that devel- oped submicronized formulation of budesonide can be used for pulmonary drug delivery for high deposition to deep lungs tissues

Regional growth and energy supply: Is there an energy security issue?

This study examines how the growth of the developing world might affect energy markets in the future. Based on recent growth trends, world energy demand could reasonably be expected to grow from about 350 Exajoules (EJ: 1.0E18=0.95 Quad) to nearly 1025 EJ by the year 2020, nearly 3x current consumption estimates. Introduction of more energy-efficient technologies could reduce this growth by about 17% to 830 EJ. But one cannot rely exclusively on current trends to forecast future energy demand. The growth of the developing world will interact with supply to affect prices, which in turn will mitigate the growth of demand, and growth rates of energy use will be much more modes. Under the Business as Usual scenario, energy demand will grow to 835 EJ by 2020, and this could be reduced a further 15% to 714 EJ through the adoption of more energy efficient technologies. Fuel prices based on model results are analyzed. Energy security implications of rapid growth in the developing world are considered and found to be of likely little significance

Diversity of SCCmec Elements in Methicillin-Resistant Coagulase-Negative Staphylococci Clinical Isolates

in MR-CoNS can generate useful information on the mobilization and evolution of this element.). in MR-CoNS

Prediction of Opioid-Induced Respiratory Depression on Inpatient Wards Using Continuous Capnography and Oximetry: An International Prospective, Observational Trial

Background: Opioid-related adverse events are a serious problem in hospitalized patients. Little is known about patients who are likely to experience opioid-induced respiratory depression events on the general care floor and may benefit from improved monitoring and early intervention. The trial objective was to derive and validate a risk prediction tool for respiratory depression in patients receiving opioids, as detected by continuous pulse oximetry and capnography monitoring. Methods: PRediction of Opioid-induced respiratory Depression In patients monitored by capnoGraphY (PRODIGY) was a prospective, observational trial of blinded continuous capnography and oximetry conducted at 16 sites in the United States, Europe, and Asia. Vital signs were intermittently monitored per standard of care. A total of 1335 patients receiving parenteral opioids and continuously monitored on the general care floor were included in the analysis. A respiratory depression episode was defined as respiratory rate ≤5 breaths/min (bpm), oxygen saturation ≤85%, or end-tidal carbon dioxide ≤15 or ≥60 mm Hg for ≥3 minutes; apnea episode lasting >30 seconds; or any respiratory opioid-related adverse event. A risk prediction tool was derived using a multivariable logistic regression model of 46 a priori defined risk factors with stepwise selection and was internally validated by bootstrapping. Results: One or more respiratory depression episodes were detected in 614 (46%) of 1335 general care floor patients (43% male; mean age, 58 ± 14 years) continuously monitored for a median of 24 hours (interquartile range [IQR], 17-26). A multivariable respiratory depression prediction model with area under the curve of 0.740 was developed using 5 independent variables: age ≥60 (in decades), sex, opioid naivety, sleep disorders, and chronic heart failure. The PRODIGY risk prediction tool showed significant separation between patients with and without respiratory depression (P < .001) and an odds ratio of 6.07 (95% confidence interval [CI], 4.44-8.30; P < .001) between the high- and low-risk groups. Compared to patients without respiratory depression episodes, mean hospital length of stay was 3 days longer in patients with ≥1 respiratory depression episode (10.5 ± 10.8 vs 7.7 ± 7.8 days; P < .0001) identified using continuous oximetry and capnography monitoring. Conclusions: A PRODIGY risk prediction model, derived from continuous oximetry and capnography, accurately predicts respiratory depression episodes in patients receiving opioids on the general care floor. Implementation of the PRODIGY score to determine the need for continuous monitoring may be a first step to reduce the incidence and consequences of respiratory compromise in patients receiving opioids on the general care floor

Promotion of Intestinal Peristalsis by Bifidobacterium spp. Capable of Hydrolysing Sennosides in Mice

BACKGROUND:While there are a variety of identifiable causes of constipation, even idiopathic constipation has different possible mechanisms. Sennosides, the main laxative constituents of Daio, an ancient Kampo medicine, are prodrugs that are converted to an active principle, rheinanthrone, by intestinal microbiota. In this study, we aimed to determine the sennoside hydrolysis ability of lactic acid bacterial strains and bifidobacteria in the intestine and to investigate their effect on intestinal peristalsis in mice. METHODOLOGY/PRINCIPAL FINDINGS:A total of 88 lactic acid bacterial strains and 47 bifidobacterial strains were evaluated for their ability to hydrolyze sennosides. Our results revealed that 4 strains, all belonging to the genus Bifidobacterium, had strong sennoside hydrolysis ability, exhibiting a decrease of >70% of sennoside content. By thin-layer chromatography analysis, rheinanthrone was detected in the medium cultured with B. pseudocatenulatum LKM10070 and B. animalis subsp. lactis LKM512. The fecal sennoside contents significantly (P<0.001) decreased upon oral administration of these strains as compared with the control. Intestinal peristalsis activity was measured by the moved distance of the charcoal powder administered orally. The distance travelled by the charcoal powder in LKM512-treated mice was significantly longer than that of control (P<0.05). Intestinal microbiota were analysed by real-time PCR and terminal-restriction fragment length polymorphism. The diversity of the intestinal microbiota was reduced by kanamycin treatment and the diversity was not recovered by LKM512 treatment. CONCLUSION/SIGNIFICANCE:We demonstrated that intestinal peristalsis was promoted by rheinanthrone produced by hydrolysis of sennoside by strain LKM512 and LKM10070

Reprogramming of Escherichia coli K-12 Metabolism during the Initial Phase of Transition from an Anaerobic to a Micro-Aerobic Environment

Background: Many bacteria undergo transitions between environments with differing O2 availabilities as part of their natural lifestyles and during biotechnological processes. However, the dynamics of adaptation when bacteria experience changes in O2 availability are understudied. The model bacterium and facultative anaerobe Escherichia coli K-12 provides an ideal system for exploring this process. Methods and Findings: Time-resolved transcript profiles of E. coli K-12 during the initial phase of transition from anaerobic to micro-aerobic conditions revealed a reprogramming of gene expression consistent with a switch from fermentative to respiratory metabolism. The changes in transcript abundance were matched by changes in the abundances of selected central metabolic proteins. A probabilistic state space model was used to infer the activities of two key regulators, FNR (O2 sensing) and PdhR (pyruvate sensing). The model implied that both regulators were rapidly inactivated during the transition from an anaerobic to a micro-aerobic environment. Analysis of the external metabolome and protein levels suggested that the cultures transit through different physiological states during the process of adaptation, characterized by the rapid inactivation of pyruvate formate-lyase (PFL), a slower induction of pyruvate dehydrogenase complex (PDHC) activity and transient excretion of pyruvate, consistent with the predicted inactivation of PdhR and FNR. Conclusion: Perturbation of anaerobic steady-state cultures by introduction of a limited supply of O2 combined with time-resolved transcript, protein and metabolite profiling, and probabilistic modeling has revealed that pyruvate (sensed by PdhR) is a key metabolic signal in coordinating the reprogramming of E. coli K-12 gene expression by working alongside the O2 sensor FNR during transition from anaerobic to micro-aerobic conditions

Analysis of obstetric complications and uterine connective tissue in tenascin-X-deficient humans and mice

Tenascin-X (TNX) is a large, multi-domain, extracellular matrix glycoprotein. Complete deficiency of TNX in humans leads to a recessive form of Ehlers-Danlos syndrome (EDS), and TNX haploinsufficiency is a cause of hypermobility type EDS. EDS patients appear to have a higher risk of several complications during pregnancy, such as pelvic instability, premature rupture of membranes, and postpartum hemorrhage. Here, we present a study of genitourinary and obstetric complications in TNX-deficient women of reproductive age. We have found complications, such as uterus prolapses, that are in agreement with previous findings in other EDS types. In TNX knockout (KO) mice, we have observed mild pregnancy-related abnormalities. Morphological and immunohistological analysis of uterine tissues has not revealed obvious quantitative or spatial differences between TNX KO and wildtype mice with respect to collagen types I, III, V, and XII or elastic fibers. We conclude that TNX-deficient women are at risk of obstetric complications, but that TNX KO mice show only a mild phenotype. Furthermore, we show that TNX is involved in the stability of elastic fibers rather than in their initial deposition

Genetically Engineered Alginate Lyase-PEG Conjugates Exhibit Enhanced Catalytic Function and Reduced Immunoreactivity

Alginate lyase enzymes represent prospective biotherapeutic agents for treating bacterial infections, particularly in the cystic fibrosis airway. To effectively deimmunize one therapeutic candidate while maintaining high level catalytic proficiency, a combined genetic engineering-PEGylation strategy was implemented. Rationally designed, site-specific PEGylation variants were constructed by orthogonal maleimide-thiol coupling chemistry. In contrast to random PEGylation of the enzyme by NHS-ester mediated chemistry, controlled mono-PEGylation of A1-III alginate lyase produced a conjugate that maintained wild type levels of activity towards a model substrate. Significantly, the PEGylated variant exhibited enhanced solution phase kinetics with bacterial alginate, the ultimate therapeutic target. The immunoreactivity of the PEGylated enzyme was compared to a wild type control using in vitro binding studies with both enzyme-specific antibodies, from immunized New Zealand white rabbits, and a single chain antibody library, derived from a human volunteer. In both cases, the PEGylated enzyme was found to be substantially less immunoreactive. Underscoring the enzyme's potential for practical utility, >90% of adherent, mucoid, Pseudomonas aeruginosa biofilms were removed from abiotic surfaces following a one hour treatment with the PEGylated variant, whereas the wild type enzyme removed only 75% of biofilms in parallel studies. In aggregate, these results demonstrate that site-specific mono-PEGylation of genetically engineered A1-III alginate lyase yielded an enzyme with enhanced performance relative to therapeutically relevant metrics.Cystic Fibrosis Foundation (Research Development Program)National Center for Research Resources (U.S.) (P20RR018787-06

In-Vitro Activity of Polymyxin B, Rifampicin, Tigecycline Alone and in Combination against Carbapenem-Resistant Acinetobacter baumannii in Singapore

OBJECTIVE: Carbapenem-resistant Acinetobacter baumannii (CR-AB) is an emerging cause of nosocomial infections worldwide. Combination therapy may be the only viable option until new antibiotics become available. The objective of this study is to identify potential antimicrobial combinations against CR-AB isolated from our local hospitals. METHODS: AB isolates from all public hospitals in Singapore were systematically collected between 2006 and 2007. MICs were determined according to CLSI guidelines. All CR-AB isolates were genotyped using a PCR-based method. Clonal relationship was elucidated. Time-kill studies (TKS) were conducted with polymyxin B, rifampicin and tigecycline alone and in combination using clinically relevant (achievable) unbound concentrations. RESULTS: 31 CR AB isolates were identified. They are multidrug-resistant, but are susceptible to polymyxin B. From clonal typing, 8 clonal groups were identified and 11 isolates exhibited clonal diversity. In single TKS, polymyxin B, rifampicin and tigecycline alone did not exhibit bactericidal activity at 24 hours. In combination TKS, polymyxin plus rifampicin, polymyxin B plus tigecycline and tigecycline plus rifampicin exhibited bactericidal killing in 13/31, 9/31 and 7/31 isolates respectively at 24 hours. Within a clonal group, there may be no consensus with the types of antibiotics combinations that could still kill effectively. CONCLUSION: Monotherapy with polymyxin B may not be adequate against polymyxin B susceptible AB isolates. These findings demonstrate that in-vitro synergy of antibiotic combinations in CR AB may be strain dependant. It may guide us in choosing a pre-emptive therapy for CR AB infections and warrants further investigations