A STUDY OF IMMUNOHISTOCHEMICAL EXPRESSION OF GALECTIN-3 IN NON-NEOPLASTIC AND NEOPLASTIC LESIONS OF THYROID

Introduction: Thyroid lesions are fairly common and have a wide spectrum of diseases ranging from functional, immune mediated to neoplastic lesions. Malignancy of thyroid constitutes approximately 1% of all cancers. These malignant tumors of thyroid gland exhibit a variety of histopathologies and clinical behavior. Immune markers are gaining more and more importance in diagnostic pathology, especially in the differential diagnostics and in the grading of thyroid gland tumors. In the recent times Galectin-3 has received notable recognition for its usefulness as a diagnostic marker for thyroid cancer. To evaluate the diagnostic role of galectin-3 (Gal-3) in Aims and objectives: differentiating malignant from benign thyroid neoplasm. In this observational study, we evaluated Gal-Material and method: 3 expression in a spectrum of all non-neoplastic and neoplastic lesions including benign and malignant lesions of thyroid gland. All types of thyroidectomy specimens were xed in 10% formal saline and subjected to histopathological examination. Sections were stained with H&E stain. Gal-3 immunoperoxidase reaction was carried out in histological sections from all the cases and descriptive analysis was done. In the study of 100 cases of thyroid swelling were included and evaluated by Result: histopathological and by using Gal-3 immunoperoxidase marker. Out of 100 cases, majority of thyroid lesions were seen in females with 76 cases (76%) while only 24 cases (24%) were seen in male. Among 66 non-neoplastic cases, only 6 cases (9.1%) and among 34 neoplastic lesions, 28 cases (82.4%) were positive for galectin-3 marker. Statistical signicance of galectin-3 expression between non-neoplastic and neoplastic thyroid lesions found to be statistically signicant (P value <0000.1). The sensitivity and specicity of galectin-3 detection of malignant lesions were found to be 82.4% and 91% respectively with 82.4% positive predictive value and 90.9% negative predictive value. We suggest that Galectin-3 expression is helpful in Conclusion: enabling better diagnosis and patient care by guiding appropriate therapeutic decisions.

Mycobacterium bovis induced human tuberculosis in India: Current status, challenges & opportunities

Tuberculosis (TB) caused by Mycobacterium tuberculosis is a leading cause of human deaths due to any infectious disease worldwide. However, infection of Mycobacterium bovis, primarily an animal pathogen, also leads to the development of ‘human tuberculosis’. Infected animals have been considered the major source of M. bovis infection and humans get exposed to M. bovis through close contact with infected animals or consumption of contaminated milk, unpasteurized dairy products and improperly cooked contaminated meat. The information on the global distribution of bovine TB (bTB) is limited, but the disease has been reported from all the livestock-producing middle- and low-income countries of the world. In recent years, there is a renewed interest for the control of bTB to minimize human infection worldwide. In India, while the sporadic presence of M. bovis has been reported in domestic animals, animal-derived food products and human beings from different geographical regions of the country, the information on the national prevalence of bTB and transmission dynamics of zoonotic TB is, however, not available. The present article reviewed published information on the status of M. bovis-induced zoonotic TB to highlight the key challenges and opportunities for intervention to minimize the risk of M. bovis infection in humans and secure optimum animal productivity in India.

Incidence & outcomes of clinically significant bleeding events in critically ill COVID-19 patients receiving Therapeutic dose AntiCoagulanTs: A retrospective cohort study (INTerACT study)

Background & objectives: The high mortality associated with the thrombotic events in hospitalized COVID-19 patients resulted in the usage of anticoagulants in varying doses. Whether high-dose anticoagulants have led to better outcomes or higher incidence of clinically significant bleeding events is debatable. Thus, this study was conducted to find the incidence of clinically significant bleeding events in moderate-to-severe COVID-19 ARDS (acute respiratory distress syndrome) patients on therapeutic anticoagulation and their outcomes. Methods: In this retrospective, single-centre study of 155 critically ill COVID-19 patients, the incidence of clinically significant bleeding was observed. Multivariate regression models were used to evaluate the association between anticoagulant regimen, coagulation and inflammatory markers with the incidence of bleeding and thrombotic events. Results: The incidence of clinically relevant non-major bleeding was 33.54 per cent (26.17-41.46%) and major bleeding was 9.03 per cent (5.02-14.69%). The anticoagulation intensity at baseline had a high odds of major bleeding when enoxaparin and dual antiplatelet therapy were used together [adjusted odds ratio OR of 434.09 (3.81-49502.95), P<0.05]. At admission, bleeders had a poorer PaO2/FiO2 ratio with more patients on invasive ventilation. At the time of bleeding, the bleeders had a higher D-dimer, ferritin, C-reactive protein and procalcitonin compared to non-bleeders. The subhazard ratio for death in bleeders was 3.35 (95% confidence interval, 1.97-5.65; P<0.001). Interpretation & conclusions: The incidence of bleeding in critically ill COVID-19 patients on therapeutic anticoagulation may increase with the severity of the disease as well as with concurrent use of dual antiplatelets. Major bleeding may also contribute to higher mortality.

Reporting adverse events of ChAdOx1 nCoV-19 coronavirus vaccine (Recombinant) among the vaccinated healthcare professionals: A cross-sectional survey

Background & objectives: The safety of the ChAdOx1 nCoV-19 vaccine is a cause of concern for many who have been vaccinated. The people have multiple concerns and fear regarding the adverse events of the vaccine. Thus, this study was undertaken to establish the safety profile of ChAdOx1 nCoV-19 Corona Virus Vaccine (Recombinant) among the healthcare professionals. Methods: This was a descriptive cross-sectional survey. After taking clearance from the institutional ethics committee 1500 healthcare professionals, who had their vaccination in the past two weeks were selected. They were provided with an online survey proforma regarding adverse events following immunization (AEFIs) of COVID-19 vaccine developed using google forms with an informed consent form affixed to it. Results: A total of 1036 individuals participated in the study. The mean and median (inter quartile range) age of the participants was 37.7 ±11.25 and 35 (29-46) yr, respectively. Of these, 52.1 per cent were female, 29.3 per cent were doctors, 33.4 per cent were nurses and 9.5 per cent were paramedical staff. Forty six per cent participants experienced one or more minor AEFIs such as pain, tenderness, redness, etc. at the injection site. Fatigue (31.75%), generalized feeling of unwell (28.57%), muscle pain (23.16%) and fever (21.71%) were the most commonly reported systemic AEFIs followed by headache (20.07%), dizziness (10.03%) and joint pains (15.25%). Most of them experienced these AEFIs within 24 h of the first dose of administration. About 42 per cent of the participants took oral antipyretics/analgesics for managing the AEFIs. Interpretation & conclusions: ChAdOx1 nCoV-19 Corona Virus Vaccine was found to be associated with mild local and systemic AEFIs that were more common after the first dose as compared to the second dose. There adverse events could be dealt with oral over-the-counter medications, with no requirement of hospitalization

Evaluating substance use in an urbanizing town of mid hills of Northern India

Background: Substance use is emerging as a major cause of morbidity and mortality across the world. Solan, a fast urbanizing town of India has witnessed mushrooming of industries and educational institutes. A surge in the persons booked under the Narcotic Drug and Psychoactive Substance Act 1985 led us to look into the determinants of the substance use in this region.Methods: We undertook a cross sectional study of one year secondary data analysis of 750 substance users screened at the de-addiction centre of Solan Hospital. The data mining was done by the cluster analysis technique. SPSS 16 and STATA 13 software were employed.Results: Mean age of users was 31 years with dominance of males (89.20 %), two third of total users were married, 75% were unemployed, 42% had upper school level education. About 60 and 38% were using cannabis and chitta (a synthetic opioid) respectively. Only 2% were consuming tobacco and alcohol. 62% of substance users had the fear of legal action and 44% had no family history of substance use. 39% had only single parent, 54% had started substance use under peer pressure and duration of use varied between 6 to 24 months.  Alcohol and cannabis were used more in urban and rural areas respectively. 63 and 70% had family history and experience of peer pressure respectively.Conclusions: Cluster analysis has generated substance specific socio-demographic determinants of substance use which would help in planning appropriate substance use alleviation strategies.

Epidermal growth factor receptor-activating mutation(E746_T751>VP) in pancreatic ductal adenocarcinoma responds to erlotinib, followed by epidermal growth factor receptor resistance-mediating mutation (A647T): A case report and literature review

Despite recent advances in treatment with multidrug chemotherapy regimens, outcomes of patients with advanced pancreatic ductal adenocarcinoma (PDAC) remain very poor. Treatment with targeted therapies has shown marginal benefits due to intrinsic or acquired resistance. Actionable mutations, while detected infrequently in patients with PDAC, are becoming increasingly used in personalized medicine. Here, we describe an epidermal growth factor receptor (EGFR)-activating mutation (E746_T751>VP) to erlotinib, a first-generation tyrosine kinase inhibitor (TKI), in a patient with metastatic PDAC. After an initial partial response to erlotinib for 12 months, the patient's disease progressed with emergence of the EGFR A647T mutation. Certainly, the patient also progressed after switching therapy to a third-generation EGFR TKI (osimertinib). This case illustrates the posttreatment evolution of EGFR A647T-mediated resistance to the first- and third-generation TKIs. To our knowledge, this is the first case to report the aforementioned activating and resistance-mediated mutations. In summary, genomic analysis performed in this patient with PDAC on the tumor biopsy and peripheral blood provided tools to understand mechanisms of response and resistance to targeted therapy with EFGR TKIs

Hemophagocytic lymphohistiocytosis: a rare and fatal manifestation of dengue fever

Authors describe a rare case of dengue fever manifesting as hemophagocytic lymphohistiocytosis. A 26-year-old man presented with acute gastroenteritis along with high grade fever, leukopenia, thrombocytopenia, acute kidney injury, liver dysfunction. Further work-up revealed elevated serum ferritin and LDH levels, and bone marrow biopsy showed hemophagocytes. As dengue fever is in rising trend all over the world, especially in Asian countries, clinicians should look out for this rare but potentially fatal complication of dengue fever.

Spectrum of Chronic Kidney Disease-Mineral Bone Disorders in Newly Detected Advanced Renal Failure Elderly Patients (CKD Stage 4 and 5)

Introduction: CKD MBD remains a complex issue in elderlypatients that has yet to be clearly defined. We aimed to evaluatethe disturbances in mineral bone disease in newly detected,untreated stage 4 and 5 elderly chronic kidney disease patients.Material and Methods: A cross-sectional observational studywith total of 93 newly detected patients underwent clinicalevaluation, biochemical assessment [serum albumin, calcium,intact parathyroid hormone(iPTH), 25- hydroxyvitamin D,phosphorus, alkaline phosphatase(ALP), creatinine], BMDmeasurement by dual-energy X-ray absorptiometry(DXA)and Lateral Abdominal X ray for aortic calcification(AAC).Results: Symptoms related to CKD-mineral bone disorderwere seen in 33.6% of the study patients. Prevalence ofhypocalcemia, hyperphosphatemia, hyperparathyroidism,and hypovitaminosis D were 64.2%, 81.1%, 49.5%,and 89.5%, respectively. Prevalence and severity ofhyperphosphatemia, hyperparathyroidism, hypocalcemia andraised ALP increases from stage 4 to stage 5 CKD, whilehypovaitaminosis D is equally prevalent in both stages.Secondary hyperparathyroidism is most common form ofCKD MBD in untreated elderly CKD population. BMD byDXA showed a low bone mass in 26.81% of our patients atdistal forearm. Patients older than 75 years more commonlyhad osteoporosis, lower ALP, phosphorus and iPTH. AAC wasseen in 13.98% of study group. Patients with AAC had higherphosphorus, iPTH and ALP. Compared to non diabetic CKDpatients, lower levels of phosphorus, ALP and iPTH wereobserved in diabetic CKD patients.Conclusion: Our study shows CKD MBD is prevalent inelderly population where symptoms alone are not enough todiagnose the bone disease.

Acanthosis Nigricans: A Marker Of Metabolic Syndrome Among Obese Children

xThis was a prospective case controlled study to determine the association of Acanthosis Nigricans (AN) and Metabolic Syndrome (MS) in obese children 10–16 years of age. We recruited 50 obese children with AN (cases) and 50 obese children without AN (controls) attending the Pediatric OPD from May 2013 to August 2014 after obtaining informed consent from their parents. Children were defined as obese if their BMI was ≥ 95th percentile for age and sex (CDC). International Diabetes Federation (IDF) 2007 criteria were used to define MS. Detailed history, physical examination, anthropometry and blood sampling were done in each case. Samples for High Density Lipoprotein Cholesterol (HDL-C), Fasting Plasma Glucose (FPG) and Triglycerides (TG) were evaluated at the in-hospital NABL certified SRL laboratory. MS was found in 64% of obese children with AN as compared to 30% of obese children without AN which was highly significant. Therefore, AN is a significant marker of MS.

Concurrent hepatitis B and autoimmune hepatitis a rare presentation of acute liver failure: a case report

Acute liver failure with concurrent Hepatitis b and autoimmune hepatitis is an extremely rare case. We report a 25 year female presenting to our emergency with fatigue and somnolence, distention of abdomen, jaundice, melena and increased hair loss leading to alopecia. Physical examination revealed jaundice, an enlarged liver, ascites and tenderness of upper right abdomen. Laboratory tests revealed an increased level, bilirubin, GGT, increased INR and prothrombin time with elevated IgG levels, and the presence of anti-smooth muscle antibodies, Anti-nuclear antibodies and HBV infection markers. The patient was diagnosed with liver failure resulting from chronic active hepatitis B with an autoimmune component. The treatment consisted of steroids, azathioprine, vitamin K, low-protein diet and lactulose enemas. After undergoing a molecular test (HBV DNA 3.23 × 10 IU/mL and HBeAg reactive), the treatment was modified by adding tenofovir disoproxil fumarate. After one month the patient was discharged in good clinical condition, with the recommendation of continued tenofovir disoproxil fumarate and prednisone. In subsequent follow-ups, no clinical deterioration or abnormal biochemical liver function test results were found.

Ages and Stages Questionnaire: a global screening scale / Ages and Stages Questionnaire: una escala de evaluación global

Abstract: With standardized screening tools, research studies have shown that developmental disabilities can be detected reliably and with validity in children as young as 4 months of age by using the instruments such as the Ages and Stages Questionnaire. In this review, we will focus on one tool, the Ages and Stages Questionnaire, to illustrate the usefulness of developmental screening across the globe.

Resumen: Mediante el uso de herramientas de evaluación estandarizada, algunos estudios de investigación han demostrado que discapacidades de desarrollo se pueden detectar con fiabilidad y validez en niños desde los 4 meses de edad mediante el uso de los instrumentos estandarizados como el Ages and Stages Questionnaire (Cuestionario de las Edades y Etapas). Para ilustrar la utilidad de la evaluación del desarrollo infantil a escala global, en este trabajo se revisará la herramienta Ages and Stages Questionnaire.

Early detection of multidrug resistant (MDR) Mycobacterium tuberculosis in a single tube with in-house designed fluorescence resonance energy transfer (FRET) probes using real-time PCR.

Rapid and correct diagnosis is crucial for the management of multidrug resistance (MDR) in Mycobacterium tuberculosis (MTB). The present study aims at rapid diagnosis for identification of multidrug resistance tuberculosis (MDR-TB) using real-time PCR. FRET hybridization probes targeting most prominent four selected codons for rpoB526 and 531 and for katG314 and 315 genes were designed and evaluated on 143 clinical MTB isolates and paired sputa for rapid detection of MDR-TB. The results of real-time PCR were compared with gold standard L-J proportion method and further validated by DNA sequencing. Of the 143 MTB positive cultures, 85 and 58 isolates were found to be ‘MDR’ and ‘pan susceptible’, respectively by proportion L-J method. The sensitivity of real-time PCR for the detection of rifampicin (RIF) and isoniazid (INH) were 85.88 and 94.11%, respectively, and the specificity of method was found to be 98.27%. DNA sequencing of 31 MTB isolates having distinct melting temperature (Tm) as compared to the standard drug susceptible H37Rv strain showed 100% concordance with real-time PCR results. DNA sequencing revealed the mutations at Ser531Leu, His526Asp of rpoB gene and Ser315Thr, Thr314Pro of katG gene in RIF and INH resistance cases. This real-time PCR assay that targets limited number of loci in a selected range ensures direct and rapid detection of MDR-TB in Indian settings. However, future studies for revalidation as well as refinement are required to break the limitations of MDR-TB detection.

In vitro validation of self designed “universal human Influenza A siRNA”.

The genomic variability of Influenza A virus (IAV) makes it difficult for the existing vaccines or anti-influenza drugs to control. The siRNA targeting viral gene induces RNAi mechanism in the host and silent the gene by cleaving mRNA. In this study, we developed an universal siRNA and validated its efficiency in vitro. The siRNA was designed rationally, targeting the most conserved region (delineated with the help of multiple sequence alignment) of M gene of IAV strains. Three level screening method was adopted, and the most efficient one was selected on the basis of its unique position in the conserved region. The siRNA efficacy was confirmed in vitro with the Madin Darby Canine Kidney (MDCK) cell line for IAV propagation using two clinical isolates i.e., Influenza A/H3N2 and Influenza A/pdmH1N1. Of the total 168 strains worldwide and 33 strains from India, 97 bp long (position 137-233) conserved region was identified. The longest ORF of matrix gene was targeted by the selected siRNA, which showed 73.6% inhibition in replication of Influenza A/pdmH1N1 and 62.1% inhibition in replication of Influenza A/H3N2 at 48 h post infection on MDCK cell line. This study provides a basis for the development of siRNA which can be used as universal anti-IAV therapeutic agent.

Application of IS1311 locus 2 PCR-REA assay for the specific detection of ‘Bison type’ Mycobacterium avium subspecies paratuberculosis isolates of Indian origin.

Background & objectives: Of the three major genotypes of Mycobacterium avium subspecies paratuberculosis (MAP), ‘Bison type’ is most prevalent genotype in the domestic livestock species of the country, and has also been recovered from patients suffering from Crohn’s disease. Recently, a new assay based on IS1311 locus 2 PCR- restriction endonuclease analysis (REA) was designed to distinguish between ‘Indian Bison type’ and non-Indian genotypes. The present study investigated discriminatory potential of this new assay while screening of a panel of MAP isolates of diverse genotypes and from different geographical regions. Methods: A total of 53 mycobacterial isolates (41 MAP and 12 mycobacterium other than MAP), three MAP genomic DNA and 36 MAP positive faecal DNA samples from different livestock species (cattle, buffaloes, goat, sheep and bison) and geographical regions (India, Canada, USA, Spain and Portugal) were included in the study. The extracted DNA samples (n=92) were analyzed for the presence of MAP specific sequences (IS900, ISMav 2 and HspX) using PCR. DNA samples were further subjected to genotype differentiation using IS1311 PCR-REA and IS1311 L2 PCR-REA methods. Results: All the DNA samples (except DNA from non-MAP mycobacterial isolates) were positive for all the three MAP specific sequences based PCRs. IS1311 PCR-REA showed that MAP DNA samples of Indian origin belonged to ‘Bison type’. Whereas, of the total 19 non-Indian MAP DNA samples, 2, 15 and 2 were genotyped as ‘Bison type’, ‘Cattle type’ and ‘Sheep type’, respectively. IS1311 L2 PCR-REA method showed different restriction profiles of ‘Bison type’ genotype as compared to non-Indian DNA samples. Interpretation & conclusions: IS1311 L2 PCR-REA method successfully discriminated ‘Indian Bison type’ from other non-Indian genotypes and showed potential to be future epidemiological tool and for genotyping of MAP isolates.

A Comparative Evaluation of the Laryngeal Mask Airway Proseal and Tracheal Intubation for Laparoscopic Cholecystectomy.

The study evaluated 60 adult patients of either sex belonging to ASA grade I & II, who were randomly allocated into 2 groups of 30 each; Group I-Proseal LMA group and Group II -ETT group. They were premedicated with inj. glycopyrrolate 5-10 mcg/kg and inj.butarphanol 2mg intramuscular 45 minutes prior to surgery. Patients were preoxygenated & anaesthesia was induced using oxygen + inj. propofol (2mg/kg) + isoflurane + inj. vecuronium bromide (0.1 mg/kg). PLMA or ETT was inserted as per group. Insertion characteristics i.e. ease of insertion and insertion time were noted. A 16Fr gastric tube was passed into the stomach in every patient and connected to continuous suction. Anesthesia was maintained with nitrous oxide, oxygen, isoflurane and inj. vecuronium bromide. Ventilation was set at 10 ml/kg and 15 breaths/min.The demographic data of both the groups were comparable. The mean age in this study was 42.2 years (22 females and 8 males) in Group I (PLMA) and 40.8 years (23 females and 7 males) in Group II (ETT). The mean BMI of patients in Group I was 22.7 kg/m2 and in Group II was 23.1 kg/m2. Insertion of device was graded as easy in 93% of patients in PLMA group and 96% of patients in ETT group. Mean time taken for successful placement of device in PLMA group was 15.83 s and in ETT group was 17.1 s, which was, however, statistically insignificant (p value = 0.095).The insertion of NGT through drain tube of PLMA was easier than via nose in ETT group. The mean insertion time taken to insert NGT through PLMA was significantly less (10.03 s) than via nose (12.7 s) in intubated patients.The anesthetic and peritoneal insufflation times in both groups were comparable. . The ventilation was adequate to maintain Sp02 of 99% - 100% in both the groups. Based on the study ,it may be said that ,Proseal laryngeal mask airway seemed to be a safe and effective alternative to endotracheal intubation in patients of laparoscopic cholecystectomy.

Type 2 diabetes mellitus affects male fertility potential.

Objective: Diabetes is a syndrome that affects all the physiological systems of the body, therefore this study was undertaken to compare the seminogram parameters in diabetics and non- diabetics. Study design: The study was carried out at Male Infertility and Reproductive Physiology unit in the Department of Physiology, MGIMS, Sevagram, Wardha. 25 normozoospermic subjects with type 2 diabetes and 25 normozoospermic non diabetic subjects were recruited in the study. The semen samples were analyzed for sperm concentration, motility and morphology. Results: In diabetic group the sperm concentration was 24.6 millions/ml with the motility of 52.3% and normal morphology 31.5%, while in non-diabetic group the sperm concentration was 42.7 millions/ml with 63.1% motility and 47.2% normal morphology. Conclusion: Thus our observations indicate that there is a detrimental effect of type 2 diabetes mellitus on semen parameters.

EFFECTS OF SINGLE, BINARY AND TERTIARY COMBINATIONS WITH Jatropha gossypifolia AND OTHER PLANT-DERIVED MOLLUSCICIDES ON REPRODUCTION AND SURVIVAL OF THE SNAIL Lymnaea acuminata / Efeitos de combinações unitárias, binárias e terciárias de Jatropha gossypifolia e outros muluscicidas derivados de plantas na reprodução e sobrevivência do caramujo Lymnaea acuminata

The effect of sub-lethal doses (40% and 80% of LC50/24h) of plant derived molluscicides of singly, binary (1:1) and tertiary (1:1:1) combinations of the Rutin, Ellagic acid, Betulin and taraxerol with J. gossypifolia latex, leaf and stem bark powder extracts and their active component on the reproduction of freshwater snail Lymnaea acuminata have been studied. It was observed that the J. gossypifolia latex, stem bark, individual leaf and their combinations with other plant derived active molluscicidal components caused a significant reduction in fecundity, hatchability and survival of young snails. It is believed that sub-lethal exposure of these molluscicides on snail reproduction is a complex process involving more than one factor in reducing the reproductive capacity.

O efeito de doses sub-letais (40% e 80% de LC50/24h) de moluscicidas derivados de plantas com combinações unitárias, binárias (1:1) e terciárias (1:1:1) de Rutin, ácido Elágico, Betulin e taraxerol com látex da J. gossypifolia, folhas e extrato em pó de casca de caule e seus componentes ativos foram estudados na reprodução do caramujo de água fresca Lymnaea acuminata. Foi observado que o látex da J. gossypifolia, casca do caule, folhas individualmente e suas combinações com componentes moluscicidas ativos derivados de outras plantas causaram redução significante na fecundidade, incubação e sobrevivência dos caramujos jovens. Acredita-se que a exposição sub-letal destes moluscicidas sobre a reprodução dos caramujos é processo complexo envolvendo mais de um fator na redução da capacidade reprodutiva.

In silico designing of siRNA targeting PB 1 gene of Influenza A virus and in vitro validation.

Genomic variability makes Influenza A virus (IAV) ‘the least susceptible’ to existing vaccines or anti-influenza drugs. siRNA targeting viral gene silents the gene by cleaving mRNA. Present study aimed to develop siRNA targeting polymerase basic 1 (PB1) gene and to validate its efficiency in vitro. siRNA was designed rationally, targeting the most conserved region of PB1 gene of IAV strains. Total 147 strains worldwide and 42 Indian strains, when aligned, showed seven sets of conserved regions (> 30 bp stretch and < 5% mismatches). To choose the most efficient siRNA, three levels screening method was developed. Finally one pair of siRNA was chosen due to its unique position in conserved region. siRNA efficacy was confirmed in vitro on Madin Darby Canine Kidney (MDCK) cell line propagating two clinical isolates i.e. Influenza A/H3N2 [A/India/LKO864/ 2011(H3N2)] and Influenza A/pandemicH1N1 [A/India/LKO2151/2012(H1N1)]. The longest ORF was targeted by the selected siRNA, which showed 57 % inhibition in replication of Influenza A/pdmH1N1 and 60.6 % inhibition in replication of Influenza A/H3N2 at 72 hpi and 48 hpi respectively on MDCK cell line. This study shows that siRNA targeting PB1 may be moderately effective in controlling IAV replication so can be used as anti-IAV therapeutic agent.

Deciphering the binding modes of hematoporphyrin to bovine serum albumin.

Interaction of proteins with small molecules is important in understanding delivery and transport of different therapeutic agents, including drugs. In the present study, we investigated the interaction between hematoporphyrin (HP), the principal component of photosensitizing drug with bovine serum albumin (BSA) in aqueous buffer solution using UV-Vis absorption spectroscopy and fluorescence measurements. The results were further substantiated by molecular docking and molecular dynamics (MD) simulation. Our results revealed that fluorescence of BSA was dominantly quenched by the ground-state complex formation with HP accompanied by the electronic energy transfer (EET) to the later. We experimentally determined the thermodynamic parameters such as G0, H0, and S0 for the HP-BSA system which were -35.5 kJ mole-1, -56.4 kJ mole-1 and -0.06 kJ mole-1 K-1, respectively. These parameters suggested hydrogen-bonding and Van der Waals forces playing major role in the complexation. This was also supported by the binding energy parameters calculated by molecular docking. Moreover, the experimentally determined G0 nicely correlated with those determined by molecular docking and MD-simulation. Further, computational results clearly showed that the binding of HP with BSA in the subdomains IB and IIA.

Influenza virus genotypes circulating in and around Lucknow, Uttar Pradesh, India, during post pandemic period, August 2010 - September 2012.

Background & objectives: During the post influenza pandemic period, continuous surveillance of influenza virus and its subtypes is mandatory to help the policy makers to take effective and appropriate decisions. Therefore, this study was planned to determine the pattern of influenza virus activity in context to various meteorological and clinical parameters in and around Lucknow, Uttar Pradesh, India, during post pandemic period August 2010 - September 2012. Methods: Nasal swabs/throat swabs/nasopharyngeal aspirates of 2669 patients were collected. One-step real time PCR for detection of influenza virus was done according to the Centers for Disease Control and Prevention (CDC) protocol. Results: Influenza positivity was 15.8 per cent (423/2669) in symptomatic patients. Of the 423 total positives, 192 (7.2%) were influenza A and 231 (8.7%) were influenza B. Positivity for influenza virus was significantly (P=0.001, OR=2.9, CI=1.9-4.3) higher in patients with Influenza like illness (ILI) (17.4%, 396/2271) than those with severe acute respiratory illness (SARI) (6.8%, 27/398). Influenza A positive samples were subtyped as; pdmH1N1 (67.2%, 129/192) and seasonal H3N2 (32.8%, 63/192). It significantly correlated with monthly mean rainfall, humidity and dew point while atmospheric pressure was inversely related. No significant association was found with temperature and wind speed. Clinical variations were observed between different strains of Influenza virus. Interpretation & conclusions: The findings provide a clear picture of different clinical presentations of various strains of influenza A and B viruses and epidemiology of influenza infection from Lucknow (UP), India. The seasonality of influenza virus infection showed variation in relation to different environmental factors. Pandemic H1N1 caused more systemic infection than seasonal influenza A/H3N2 virus.