A novel IgY-Aptamer hybrid system for cost-effective detection of SEB and its evaluation on food and clinical samples.

In the present study, we introduce a novel hybrid sandwich-ALISA employing chicken IgY and ssDNA aptamers for the detection of staphylococcal enterotoxin B (SEB). Cloning, expression and purification of the full length recombinant SEB was carried out. Anti-SEB IgY antibodies generated by immunizing white leg-horn chickens with purified recombinant SEB protein and were purified from the immunized egg yolk. Simultaneously, ssDNA aptamers specific to the toxin were prepared by SELEX method on microtiter well plates. The sensitivity levels of both probe molecules i.e., IgY and ssDNA aptamers were evaluated. We observed that the aptamer at 250 ngmL(-1) concentration could detect the target antigen at 50 ngmL(-1) and the IgY antibodies at 250 ngmL(-1), could able to detect 100 ngmL(-1) antigen. We further combined both the probes to prepare a hybrid sandwich aptamer linked immune sorbent assay (ALISA) wherein the IgY as capturing molecule and biotinylated aptamer as revealing probe. Limit of detection (LOD) for the developed method was determined as 50 ngmL(-1). Further, developed method was evaluated with artificially SEB spiked milk and natural samples and obtained results were validated with PCR. In conclusion, developed ALISA method may provide cost-effective and robust detection of SEB from food and environmental samples.

Identification of appropriate outcome indices in head and neck cancer and factors influencing them.

Audit of early postoperative outcomes adjusted for patient case mix is still in its infancy in head and neck surgery. Nevertheless the role for audit of early postoperative outcomes is obvious. The primary outcome measure of this study was to identify factors that are associated with early mortality or morbidity after surgery for head and neck squamous cell carcinoma (HNSCC). The secondary outcome measure was to develop a pilot score that allows for risk-adjustment of outcome data to facilitate departmental audit. In this series the mortality rate was low (2.8%), in keeping with other published series. Complications, including those causing death, occurred after 38.1% of operations. Independent risk factors for mortality on logistic regression were shown to be previous HNSCC (P=0.03), estimated blood loss (l) (P=0.03), and extracapsular spread (P=0.05). Age (P=0.01), tracheostomy (P<0.01), estimated blood loss (l) (P=0.05), and duration of anaesthesia (P<0.01) were independent predictors of complications. Models predicting for risk demonstrated good discrimination (area under the curve statistics) for mortality (0.86) and morbidity (0.81). These pilot scores need external validation and may herald a means of facilitating risk-adjustment in the audit of early outcomes, allowing meaningful comparison of surgeons and their units over time.

Molecular epidemiology and complete genome characterization of H1N1pdm virus from India.

BACKGROUND: Influenza A virus is one of world's major uncontrolled pathogen, causing seasonal epidemic as well as global pandemic. This was evidenced by recent emergence and continued prevalent 2009 swine origin pandemic H1N1 Influenza A virus, provoking first true pandemic in the past 40 years. In the course of its evolution, the virus acquired many mutations and multiple unidentified molecular determinants are likely responsible for the ability of the 2009 H1N1 virus to cause increased disease severity in humans. Availability of limited data on complete genome hampers the continuous monitoring of this type of events. Outbreaks with considerable morbidity and mortality have been reported from all parts of the country. METHODS/RESULTS: Considering a large number of clinical cases of infection complete genome based sequence characterization of Indian H1N1pdm virus and their phylogenetic analysis with respect to circulating global viruses was undertaken, to reveal the phylodynamic pattern of H1N1pdm virus in India from 2009-2011. The Clade VII was observed as a major circulating clade in phylogenetic analysis. Selection pressure analysis revealed 18 positively selected sites in major surface proteins of H1N1pdm virus. CONCLUSIONS: This study clearly revealed that clade VII has been identified as recent circulating clade in India as well globally. Few clade VII specific well identified markers undergone positive selection during virus evolution. Continuous monitoring of the H1N1pdm virus is warranted to track of the virus evolution and further transmission. This study will serve as a baseline data for future surveillance and also for development of suitable therapeutics.

Transcriptional regulation and activation of the mitogen-activated protein kinase pathway after Japanese encephalitis virus infection in neuroblastoma cells.

Japanese encephalitis virus (JEV), the most frequent and the single most important cause of encephalitis worldwide, has spread throughout most of Asia. For the development of appropriate and effective therapy, there is an immediate requirement to understand the role of host factors in JEV-induced neuropathogenesis. In the present study, we investigated the role of mitogen-activated protein kinases (MAPKs) in JEV infection of mouse neuroblastoma (N2A) cells. The MAPK pathway was studied at the transcriptional level to access the gene expression profile at different time points after JEV infection. The effector MAPK genes were also analyzed for protein expression and activation. Gene expression analysis showed a significant regulation of extracellular signal-regulated kinases (ERK)1, ERK2 and c-Jun N-terminal kinase (JNK)3 genes along with their downstream transcription factors such as Mef2c, c-Jun and Sfn. Experiments with the JNK inhibitor, SP600125, and the ERK inhibitor, PD98059, showed the involvement of JNK in JEV-induced caspase-3 activation and apoptosis, but ERK1/2 had no effect. Overall, our results show the transcriptional regulation of the MAPK pathway and the essential role of JNK in JEV-induced apoptosis in neuroblastoma cells. These findings provide a new insight into the role of the mitogen- and stress-activated kinases in JEV pathogenesis and opens up new avenues of therapeutics.

Appearance of E1: A226V mutant Chikungunya virus in Coastal Karnataka, India during 2008 outbreak.

Chikungunya has resurged in the form of unprecedented explosive epidemic in 2006 after a long gap in India affecting 1.39 million of persons. The disease continued for the next two consecutive years affecting 59,535 and 64,548 persons during 2007 and 2008 respectively. The 2008 outbreak being the second largest among these three years the information regarding the etiology and the mutations involved are useful for further control measures. Among the 2008 outbreaks the Coastal Karnataka accounts for the 46,510 persons. An in-depth investigation of Chikungunya epidemic of Coastal Karnataka, India, 2008 by serology, virus isolation, RT-PCR and genome sequencing revealed the presence and continued circulation of A226V mutant Chikungunya virus. The appearance of this mutant virus was found to be associated with higher prevalence of vector Aedes albopictus and the geographical proximity of coastal Karnataka with the adjoining Kerala state. This is the first report regarding the appearance of this mutation in Karnataka state of India. The present study identified the presence and association of A226V mutant virus with Chikungunya outbreak in India during 2008.

The epidemiology and treatment of mesothelioma in South East England 1985-2002.

OBJECTIVES: To describe trends in the incidence of mesothelioma for men and women in South East England and the geographical variation at the level of primary care trust. To describe treatment patterns by cancer network of residence, and relative survival by cancer network, disease stage and treatment modality. METHODS: 5753 cases were extracted from the Thames Cancer Registry database. We calculated age standardised incidence rates for each year, age specific incidence rates in 10 year age groups, and we used linear regression to compute the average annual percentage change in age standardised incidence. We used Poisson regression to analyse generational trends in incidence. RESULTS: Men had five times higher incidence of mesothelioma than women. In men, there was an overall 4% increase per year between 1985 and 2002. Over the same period, the overall increase in incidence for women was 5% per year. The incidence was highest in men aged over 70 years, and men aged over 80 years had the highest increase of 8% per year. The incidence rate ratio increased for men born between 1892 and 1942 and started to slow for those born from 1947 onwards. Areas along the Thames and its estuary had the highest incidence. There was some variation by cancer network in the proportion of patients receiving cancer surgery, radiotherapy and chemotherapy. There were no discernable differences in relative survival by cancer network of residence or disease stage but those receiving combined treatment had higher 5 year survival. CONCLUSIONS: Mesothelioma incidence has increased in South East England, particularly for men aged over 70 years. The highest incidence occurs along the Thames and its estuary, reflecting areas of asbestos use in shipbuilding and industry in the past. More research is needed to understand the interrelationships of prognostic factors, treatment choices and survival, and to determine the best care and support for these patients and their families.

Complications encountered in a prospective series of 182 patients treated surgically for mouth cancer.

Outcomes of surgical treatment for patients with mouth cancer can be limited by the risk of perioperative complications. This study identifies factors that predict for complications in such patients. Between 1992 and 2000, 182 patients had surgery for mouth cancer. The patient, their medical and surgical characteristics as well as perioperative complications were identified. Univariate analysis was carried out to determine which characteristics were associated with complications. Complications occurred in 85 patients (47%). Fifty-three percent of the complications were of intermediate severity and 15.6% were major. The operative death rate was 3.2%. Factors predicting complications included pre-existing cardiovascular (p<0.01) and respiratory disease (p=0.02), alcohol consumption (p<0.01), stage of disease, nature and scale of surgery, duration of surgery (p<0.01). Tracheostomies (p<0.01, OR 3.05), poor differentiation of tumour (p<0.05) and presence of extracapsular spread were predictive of complications. Patients with more complications are at increase risk of operative death or dying with head and neck cancer (p=0.04). Complications were also analysed into those that may be related to surgical technique and medical management. 113 (37%) complications were in this category. Factors influencing complications are multifactorial. Identification of risk factors allows individualised approach should improve outcome of treatment.

Japanese Encephalitis Outbreak, India, 2005.

An outbreak of viral encephalitis occurred in Gorakhpur, India, from July through November 2005. The etiologic agent was confirmed to be Japanese encephalitis virus by analyzing 326 acute-phase clinical specimens for virus-specific antibodies and viral RNA and by virus isolation. Phylogenetic analysis showed that these isolates belonged to genogroup 3.

Exposure-response relations for work related respiratory symptoms and sensitisation in a cohort exposed to alpha-amylase.

AIMS: To explore relations between exposure to fungal alpha-amylase and the risk of new work related respiratory symptoms or sensitisation. METHODS: A prospective cohort study among 300 bakers and millers was followed up for a maximum of seven years. Exposure to alpha-amylase was estimated by air measurements and questionnaires and classified into three categories. Symptoms were recorded with a self-administered questionnaire and skin sensitisation assessed using skin prick test (SPT). RESULTS: There were 36 new cases of chest symptoms, 86 of eyes/nose symptoms, and 24 of a positive SPT to alpha-amylase. There were exposure-response relations for chest and eyes/nose symptoms and for sensitisation, and a significantly increased prevalence ratio for chest symptoms in the highest exposure category. CONCLUSION: A reduction in alpha-amylase exposure is likely to reduce the risk for respiratory morbidity in bakery workers.

Perioperative complications and survival in patients with oral cancer.

This study identifies factors that predict for outcome and complications in patients with mouth cancer. Out of a total of 276 patients, one third received radiotherapy alone and the remainder surgery (182) of which 62 also received adjuvant radiotherapy. Factors predicting an adverse outcome at a univariate level were male gender, recurrent disease, no partner, co-existing systemic disease (abdomen and respiratory), alcohol intake, non Caucasian, stage of disease, scale of surgery, decreasing differentiation of tumour, increasing hospital stay and blood loss. On multivariate analysis, female gender, reduced scale of surgery, absence of recurrence, excess alcohol consumption, increased hospital stay and duration of surgery were predictive of improved survival. Complications occurred in 85 patients (47%). Predictive variables on univariate analysis were preexisting cardio-respiratory disease, alcohol consumption, stage of disease, nature and scale of surgery. The 5 year disease specific survival was 87% for stage I, 75% for stage II, 62% for stage III and 43% for stage IV with a 3.2% operative death rate.

Exposure-response relations among laboratory animal workers exposed to rats.

AIM: To explore exposure-response relations in a cohort of laboratory animal workers. METHODS: Exposure-response modelling was carried out in a cohort of 342 laboratory animal workers. Three exposure indices, divided into different exposure categories, were used in the analyses: intensity of exposure to rat urinary aeroallergen (RUA, the main allergen workers were exposed to), weekly duration of exposure to rats, and the product of the intensity and weekly duration of exposure. Outcomes studied were work related chest, eyes and nose, and skin symptoms that had started after employment at the sites, specific sensitisation, and a combination of symptoms and sensitisation. Cox proportional hazard modelling was used to explore exposure-response relations. Smoking, atopic status, age, and gender were taken into account. RESULTS: We observed the clearest exposure-response relations for the intensity of exposure to RUA and the various endpoints. No clear exposure-response relations were observed for the weekly duration of exposure or the product of the intensity and weekly duration of exposure. The strongest and clearest exposure-response relations for symptoms were observed among rat sensitised workers, while the non-sensitised workers only showed small increased risks of developing symptoms without clear exposure-response relations. Sensitised workers were almost four times more likely to go on to develop chest symptoms compared to non-sensitised workers.