FORMULATION, OPTIMIZATION, CHARACTERIZATION AND IN VIVO ANTI-ULCER ACTIVITY OF ESOMEPRAZOLE MAGNESIUM TRIHYDRATE GASTRORESISTANT MICROSPHERES

Objective: The objective of the present investigation was to prepare gastro-resistant microspheres of esomeprazole magnesium trihydrate (EMT) to prevent its degradation in the acidic environment of the stomach and enhance its bioavailability via intestinal absorption.Methods: EMT loaded gastro-resistant microspheres were prepared using hypromellose acetate succinate (HPMCAS) as the gastro-resistant polymer by ‘non-aqueous solvent evaporation' technique. A 3-factor 3 level factorial design was used to optimise EMT: HPMCAS ratio, the concentration of Span 80 and stirring speed with respect to percent entrapment efficiency and particle size. Further characterization was carried out using scanning electron microscopy (SEM), differential scanning calorimetry (DSC), In vitro release study and In vivo anti-ulcer activity.Results: Fourier transform infrared (FTIR) study indicated compatibility between drug and polymer. DSC study revealed that the drug was molecularly dispersed in the polymer. The optimised batch showed 49.63±1.23% drug entrapment and 170.12±3.36 μm particle size. SEM study showed that microspheres were spherical in shape. In vitro drug release study showed only 4.28±1.23% drug release in simulated gastric media in 2 hr and 93.46±1.20% release in simulated intestinal media after 1 hr from the optimised batch.Conclusion: Results of in vitro release studies indicated the gastro-resistant nature of the developed microspheres. In vivo anti-ulcer activity demonstrated that EMT loaded microspheres were able to significantly reduce ethanol-induced ulcer formation in rats' stomach as compared to the aqueous solution of EMT. So it can be concluded that the developed gastro-resistant microspheres of EMT prevented drug release in the stomach which would lead to a significant improvement in its bioavailability through enhanced intestinal absorptio

DEVELOPMENT AND STABILITY INDICATING HPLC METHOD FOR DAPAGLIFLOZIN IN API AND PHARMACEUTICAL DOSAGE FORM

Objective: To develop precise, accurate and reproducible stability assay method by RP-HPLC for estimation of dapagliflozin in API and pharmaceutical dosage form.Methods: The adequate separation was carried using agilent C18 (4.6 ml (millimeter)*150,5 µm (micromiter), mixture of acetonitrile: di-potassium hydrogen phosphate with pH-6.5 adjusted with OPA (40:60 %v/v) as a mobile phase with the flow rate of 1 ml/min (milliliter/minute) and the effluent was monitored at 222 nm (nanometer) using photo diode array detector. The retention time of dapagliflozin API and dapagliflozin tablet were 3.160 min (minute) and 3.067 min (minute) respectively.Results: Linearity for dapagliflozin was found in the range of 50-150µg/ml (microgram/milliliter) (R2 = 0.99) respectively. The accuracy of the present method was evaluated at 50 %, 100% and 150%. The % recoveries of dapagliflozin API and tablet were found to be in the range of 99.00–99.99 % and 98.50–99.99 % respectively. Precision studies were carried out and the relative standard deviation values were less than two. The method was found to be robust.Conclusion: The proposed method was found to be specific, accurate, precise and robust can be used for estimation of dapagliflozin in API and Pharmaceutical dosage form

A study of chronic kidney disease patients with no known risk factors coming to tertiary care hospital

Background: Chronic kidney disease (CKD) has become a major cause of morbidity and. in some parts of the world CKD incidence has increased regardless of hypertension, diabetes mellitus or metabolic syndrome. This study was done to identify the unknown factors which can be contributing to the increased incidence of CKD.Methods: It was a case control study. There were 61 cases and 50 controls. A detailed history regarding residence, occupation, addiction, drug intake, family history, diet and environmental factors was taken. The data was analysed to identify a common factor amongst the CKD patients who did not have history of any known risk factors of CKD.Results: Age of onset of CKD in 48% of cases was 5 years (30%) as compared to controls. Much more cases as compared to controls gave history of mixed diet (46% vs 26%). Much higher proportion of cases had history of heat exposure, excessive heating and sugarcane exposure (72%, 70% and 48% respectively) as compared to controls.Conclusions: This study supports the association of sugarcane exposure, heat exposure and excessive sweating with CKD and reports a changing trend of renal involvement starting at an earlier age. It highlights need of study with sufficient sample size and greater emphasis on family history, smoking, extent of heat exposure and sugarcane exposure to help identifying area of further research and guide policy making

Prion protein conversion at two distinct cellular sites precedes fibrillisation

The self-templating nature of prions plays a central role in prion pathogenesis and is associated with infectivity and transmissibility. Since propagation of proteopathic seeds has now been acknowledged a principal pathogenic process in many types of dementia, more insight into the molecular mechanism of prion replication is vital to delineate specific and common disease pathways. By employing highly discriminatory anti-PrP antibodies and conversion-tolerant PrP chimera, we here report that de novo PrP conversion and formation of fibril-like PrP aggregates are distinct in mechanistic and kinetic terms. De novo PrP conversion occurs within minutes after infection at two subcellular locations, while fibril-like PrP aggregates are formed exclusively at the plasma membrane, hours after infection. Phenotypically distinct pools of abnormal PrP at perinuclear sites and the plasma membrane show differences in N-terminal processing, aggregation state and fibril formation and are linked by exocytic transport via synaptic and large-dense core vesicles

Evaluation of immunization coverage among children aged 12-23 months in Surendranagar city

Background: Infectious diseases are a major cause of morbidity & mortality in children. One of the most cost effective & easy methods for child survival is immunization. In May 1974, the World Health Organization (WHO) officially launched a global immunization programme known as Expanded Programme of immunization (EPI) to protect all the children of the world against 6 Vaccine Preventable Diseases by the year 2000. It was later redesignated as Universal Immunization Programme (UIP) since 1985. The objectives of this study were to assess the dropout rate and primary immunization coverage of children aged 12-23 months in Surendranagar city and to know the various reasons for partially or not immunizing the child.Methods: A community-based cross-sectional study. Thirty clusters were selected out of a total of 282 blocks of Surendranagar using the cluster sampling method. Cluster sampling method was used for sample selection and the proforma designed by UNICEF was used as a study tool. Sample size was 210 children (7 Children from each cluster) of aged 12-23 months. The obtained data were analyzed using appropriate statistical tests like Z test and X2 test. Results: Out Of the 210 surveyed children, 121(57.62%) were males and 89(42.38%) were females. Immunization card was available for 69.52% of children and fully immunized were 70.47%. Coverage was highest for BCG (95.71%) followed by OPV3 (82.85%), DPT3 (79.52%) and lowest for measles (75.23%). As far as the dropout rate is concerned, it was 21.39%, 10.21%, and 9.37% for BCG to measles, DPT1 to DPT3, and OPV1 to OPV3, respectively. Amongst the various reasons main reasons for dropout or unimmunization of children were ignorance in about 64% and lack of information regarding time, place and schedule (21%).Conclusions: Improvement should focus on reducing the dropout rate from DPT2/OPV2 to DPT3/OPV3 and improving coverage of measles and also Vitamin A

Festivals and deterioration of aquatic environment: A case study of Idol immersion in Tapi River, India

In the civil society different festivals are celebrated, these are the integral part of human life and many of festivals are religious, seasonal change and culturally important. The present study elucidated environmental impact of Ganesh idol immersion on water quality of Tapi River and for this purpose water samples were collected during different durations (pre immersion, during immersion and post immersion) from selected sampling stations or idol immersion points of Tapi River. The important water quality parameters like pH, temperature, dissolved oxygen, free carbon dioxide, total hardness, total alkalinity, biological oxygen demand, chemical oxygen demand, oil & grease and total calcium were analysed for the study. Result shows that dissolved oxygen was depleted while remaining parameter were increased during the idol immersion and it was concluded that aquatic ecosystem of Tapi river was deteriorated and pollution and nutrient load were increased due to these religious activities. The celebration of festivals and these religious activities can’t stop but pollution can reduce to save the river

Accuracy of Immunofluorescence in the Diagnosis of Primary Ciliary Dyskinesia

RATIONALE The standard approach to diagnosis of primary ciliary dyskinesia (PCD) in the UK consists of assessing ciliary function by high-speed-microscopy and ultrastructure by election microscopy, but equipment and expertise is not widely available internationally. The identification of bi-allelic disease causing mutations is also diagnostic, but many disease causing genes are unknown, and testing is not widely available outside the USA. Fluorescent antibodies to ciliary proteins are used to validate research genetic studies, but diagnostic utility in this disease has not been systematically evaluated. OBJECTIVES: Determine utility of a panel of six fluorescent labelled antibodies as a diagnostic tool for PCD. METHODS: Immunofluorescent labelling of nasal brushings from a discovery cohort of 35 patients diagnosed with PCD by ciliary ultrastructure, and a diagnostic accuracy cohort of 386 patients referred with symptoms suggestive of disease. The results were compared to diagnostic outcome. MEASUREMENTS AND MAIN RESULTS: Immunofluorescence correctly identified mislocalised or absent staining in 100% of the discovery cohort. In the diagnostic cohort immunofluorescence successfully identified 22 of 25 patients with PCD and normal staining in all 252 in whom PCD was considered highly unlikely. Immunofluorescence additionally provided a result in 55% (39) of cases which were previously inconclusive. Immunofluorescence results were available within 14 days, costing $187 per sample compared to electron microscopy (27 days, cost$1452). CONCLUSIONS: Immunofluorescence is a highly specific diagnostic test for PCD, and improves the speed and availability of diagnostic testing, however, sensitivity is limited and immunofluorescence is not suitable as a stand-alone test

Defective airway intraflagellar transport underlies a combined motile and primary ciliopathy syndrome caused by IFT74 mutations

Ciliopathies are inherited disorders caused by defective cilia. Mutations affecting motile cilia usually cause the chronic muco-obstructive sinopulmonary disease primary ciliary dyskinesia (PCD) and are associated with laterality defects, while a broad spectrum of early developmental as well as degenerative syndromes arise from mutations affecting signalling of primary (non-motile) cilia. Cilia assembly and functioning requires intraflagellar transport of cargos assisted by IFT-B and IFT-A adaptor complexes. Within IFT-B, the N-termini of partner proteins IFT74 and IFT81 govern tubulin transport to build the ciliary microtubular cytoskeleton. We detected a homozygous 3 kb intragenic IFT74 deletion removing the exon 2 initiation codon and 40 N-terminal amino acids in two affected siblings. Both had clinical features of PCD with bronchiectasis, but no laterality defects. They also had retinal dysplasia and abnormal bone growth, with a narrowed thorax and short ribs, shortened long bones and digits and abnormal skull shape. This resembles short-rib thoracic dysplasia, a skeletal ciliopathy previously linked to IFT defects in primary cilia, not motile cilia. Ciliated nasal epithelial cells collected from affected individuals had reduced numbers of shortened motile cilia with disarranged microtubules, some mis-orientation of the basal feet, and disrupted cilia structural and IFT protein distributions. No full length IFT74 was expressed, only truncated forms that were consistent with N-terminal deletion and inframe translation from downstream initiation codons. In affinity purification mass spectrometry, exon 2-deleted IFT74 initiated from the nearest inframe downstream methionine 41 still interacts as part of the IFT-B complex, but only with reduced interaction levels and not with all its usual IFT-B partners. We propose that this is a hypomorphic mutation with some residual protein function retained, that gives rise to a non-lethal primary skeletal ciliopathy combined with defective motile cilia and PCD

Primary ciliary dyskinesia with normal ultrastructure:three-dimensional tomography detects absence of DNAH11

In primary ciliary dyskinesia (PCD), motile ciliary dysfunction arises from ciliary defects usually confirmed by transmission electron microscopy (TEM). In 30% of patients, such as those with DNAH11 mutations, apparently normal ultrastructure makes diagnosis difficult. Genetic analysis supports diagnosis, but may not identify definitive causal variants. Electron tomography, an extension of TEM, produces three-dimensional ultrastructural ciliary models with superior resolution to TEM. Our hypothesis is that tomography using existing patient samples will enable visualisation of DNAH11-associated ultrastructural defects. Dual axis tomograms from araldite-embedded nasal cilia were collected in 13 PCD patients with normal ultrastructure (DNAH11 n=7, HYDIN n=2, CCDC65 n=3 and DRC1 n=1) and six healthy controls, then analysed using IMOD and Chimera software. DNAH11 protein is localised to the proximal ciliary region. Within this region, electron tomography indicated a deficiency of >25% of proximal outer dynein arm volume in all patients with DNAH11 mutations (n=7) compared to other patients with PCD and normal ultrastructure (n=6) and healthy controls (n=6). DNAH11 mutations cause a shared abnormality in ciliary ultrastructure previously undetectable by TEM. Advantageously, electron tomography can be used on existing diagnostic samples and establishes a structural abnormality where ultrastructural studies were previously normal