Large scale purification and characterization of A21 deamidated variant-most prominent post translational modification (PTM) for insulins which is also widely observed in insulins pharmaceutical manufacturing and storage.

Biopharmaceutical development demands appropriate understanding of product related variants, which are formed due to post-translational modification and during downstream processing. These variants can lead to low yield, reduced biological activity, and suboptimal product quality. In addition, these variants may undergo immune reactions, henceforth need to be appropriately controlled to ensure consistent product quality and patient safety. Deamidation of insulin is the most common post-translational modification occurring in insulin and insulin analogues. AsnA21 desamido variant is also the most prominent product variant formed during human insulin manufacturing process and/or during the storage. Often, this deamidated variant is used as an impurity standard during in-process and final product analysis in the QC system. However, purification of large quantity of purified deamidated material is always being challenging due to highly similar mass, ionic, hydrophobic properties, and high structural similarity of the variant compared to the parent product. Present work demonstrates the simplified and efficient scalable process for generation of AsnA21 deamidated variant in powder form with ~96% purity. The mixed-mode property of anion exchange resin PolyQuat was utilized to purify the deamidated impurity with high recovery. Subsequent reversed-phase high performance liquid chromatography (RP-HPLC) step was introduced for concentration of product in bind elute mode. Elution pool undergone isoelectric precipitation and lyophilisation. The lyophilized product allows users for convenient use of the deamidated impurity for intended purposes. Detailed characterization by Mass spectrometry revealed deamidation is at AsnA21 and further confirmed that, structural and functional characterization as well as the biological activity of isolated variant is equivalent to insulin.

Plasma Fibrinogen: An Independent Predictor of Pediatric Appendicitis.

AIMS: Appendicitis, in spite of all the diagnostic advances, achieving an accurate and timely diagnosis of this common condition in children remains a challenge. Plasma fibrinogen (FB) is an acute inflammatory mediator and has been proposed and evaluated as an adjunct laboratory marker for improving diagnostic accuracy. The study evaluates the plasma values of Se FB along with other serum markers in pediatric appendicitis patients, to determine their diagnostic accuracy. METHODS: Prospective observational study on 120 patients between the age group of 5 and 12 years. All eligible enrolled cases underwent total leukocyte count (TLC), plasma FB, C reactive protein (CRP), neutrophil-lymphocyte ratio (NLR), absolute neutrophil count (ANC), and erythrocyte sedimentation rate on admission along with pediatric appendicitis score. Final confirmation of diagnosis and allotment of cohort was made by intra operative findings and histopathological confirmation. Two groups were defined: (1) Histopathologically confirmed acute appendicitis-Cases (2) Nonspecific abdominal pain-Controls. Laboratory results were statistically analyzed between the case and the control groups for diagnostic accuracy. RESULTS: Study reflected strong statistical significance in terms of leukocyte count, ANC, NLR, CRP, and FB levels. However, plasma FB (value above 4.02 g/L) had the highest diagnostic accuracy rate of 82.50% compared to other laboratory values (TLC-70.83%, CRP-70.00%). CONCLUSION: Plasma FB has emerged as an accurate diagnostic tool and its diagnostic accuracy is superior to all other laboratory parameter studied (TLC, CRP, NLR, and ANC). Plasma FB values above 4.02 g/L is an independent predictor of appendicitis and can help in reducing negative laparotomy in pediatric age group.

Genetic diversity in leafy mustard (Brassica juncea var. rugosa) as revealed by agro-morphological traits and SSR markers.

Leafy mustard (B. juncea var. rugosa) constitutes an important group of vegetable mustard crops in India and is mainly cultivated in home-backyard and hilly regions of Uttarakhand and some North-eastern states. In the present study, various agro-morphological traits, physiological and biochemical traits along with SSR markers were used for genetic diversity evaluation in a germplasm collection of leafy mustard. This study revealed a significant variation among 59 accessions of leafy mustard in both qualitative and quantitative agro-morphological traits indicating the accessions' promising potential for consumption purpose and for use in breeding programs. Maximum variability was recorded for leaf area elongation rate (CV = 53.12%), followed by total plant weight (TPW) (CV = 50.63%) and seed yield per plant (CV = 44.33%). In molecular analysis, 155 SSRs evaluated resulted in 482 alleles and the number of alleles varied form 1 to 8 with an average of 3.11 alleles per marker. A total of 122 (78.70%) SSRs resulted into polymorphic amplicons. PIC value varied from 0.32 to 0.77 with an average value of 0.44 per SSR locus. The unweighted neighbour-joining-based dendrogram analysis divided all the 59 accessions into two major groups on the basis of both agro-morphological traits and SSR markers, whereas, three subpopulations/subgroups were predicted by population STRUCTURE analysis. AMOVA indicated the presence of more variability within population than among population. Overall, agro-morphologically better performing and genetically diverse genotypes have been identified which could be further used as donors for leafy mustard improvement programs.

Milling interventions for the production of atta for Indian flat bread with low carbohydrate digestibility.

Study was aimed to produce atta for chapati, an Indian flat bread with low carbohydrate digestibility through different milling interventions; processing and formulating a functional ingredient mix (FM). Granulation, physico-chemical, rheological and chapati making characteristics of chakki atta, CA (control), roller mill atta (RA); RA replaced with 5, 10 and 15% FM (5, 10 and 15% RAFM) were evaluated. RA and RAFM samples showed lower water absorption, higher dough stability, pasting temperature and peak viscosity than CA. Evaluation of carbohydrate digestive profile showed differences in the pattern of carbohydrate digestibility and glucose release between the chapatis prepared from CA, RA and 10% RAFM. Rapidly available glucose (RAG), an indicator of glycemic response in vivo, was found to be lower in the 10% RAFM than CA. It can be concluded that milling interventions and compositional differences together determine the carbohydrate digestibility of the atta.

Directly observed therapy of sofosbuvir/ribavirin +/- peginterferon with minimal monitoring for the treatment of chronic hepatitis C in people with a history of drug use in Chennai, India (C-DOT).

We assessed the feasibility of field-based directly observed therapy (DOT) with minimal monitoring to deliver HCV treatment to people with a history of drug use in Chennai, India. Fifty participants were randomized 1:1 to sofosbuvir+peginterferon alfa 2a+ribavirin (SOF+PR) for 12 weeks (Arm 1) vs sofosbuvir+ribavirin (SOF+R) for 24 weeks (Arm 2). SOF+R was delivered daily at participant chosen venues and weekly peginterferon injections at the study clinic. HCV RNA testing was performed to confirm active HCV infection and sustained virologic response 12 weeks after treatment completion (SVR12). No baseline genotyping or on-treatment viral loads were performed. Median age was 46 years. All were male and 20% had significant fibrosis/cirrhosis. All self-reported history of injection drug use, 18% recent noninjection drug use and 38% alcohol dependence. Six discontinued treatment (88% completed treatment in each arm). Of 22 who completed SOF+PR, all achieved SVR12 (22/25=88%); 15 of 22 who completed SOF+R achieved SVR12 (15/25=60%; P=.05). Among those completing SOF+R, SVR12 was significantly less common in participants reporting ongoing substance use (36% vs 100%) and missed doses. Active substance use and missed doses did not impact SVR with SOF+PR. Field-based DOT of HCV therapy without real-time HCV RNA monitoring was feasible; however, achieving 100% adherence was challenging. SOF+PR appeared superior to SOF+R in achieving SVR12, even when doses were missed with no discontinuations due to side effects. Further exploration of short duration treatment with peginterferon plus direct-acting antivirals is warranted.

Effect of surfactants and hydrophilic polymers on the stability of an antihypertensive drug candesartan cilexetil: Evaluation by HPLC.

OBJECTIVES: The objective of this study is to investigate the effect of surfactants (polysorbate 80 and sodium lauryl sulphate) and hydrophilic polymers (polyvinylpyrrolidone and polyethylene glycol 6000) on the stability of candesartan cilexetil under isothermal stress conditions (100°C, 48h). METHODS: HPLC method was employed to evaluate the drug content and formation of degradation products in stress samples. Drug and degradation products were separated on Hypersil BDS C18 (250×4.6mm, 5µ) column using acetonitrile-water (pH 2.8) in the ratio of 85:15% v/v as a mobile phase. RESULT: Similar degradation behaviour of drug was observed with polyvinylpyrrolidone, polyethylene glycol 6000 and polysorbate 80; four common degradation peaks were observed at the retention time of 3.7, 4.5, 7.8 and 11minutes. One extra common degradation peak of very low intensity was also observed with polyethylene glycol 6000 and polysorbate 80 at the retention time of 4.2min. The drug was eluting at the retention time of 5.4min. In the case of sodium lauryl sulphate, two prominent degradation peaks were observed at the retention time of 3.7 and 13.25min along with few very low-intensity degradation peaks. CONCLUSION: The drug showed 41%, 64%, 72% and 98% degradation in presence of polyvinylpyrrolidone, polyethylene glycol 6000, polysorbate 80 and sodium lauryl sulphate, respectively.

Molecular identification of multi drug resistant bacteria from urinary tract infected urine samples.

Urinary tract infections (UTIs) are of great concern in both developing and developed countries all over the world. Even though the infections are more common in women and children, they are at a considerable rate in men and of all ages. The uropathogens causing the infections are spread through various routes. The treatment generally recommended by the physicians is antibiotic usage. But, most of the uropathogens have evolved antibiotic resistance mechanisms. This makes the present situation hectic in control and prevention of UTIs. The present study aims to illustrate the multidrug resistance patterns among isolated bacterial strains from infected urine samples in Odisha state, India. Four bacterial strains were isolated and identified as Proteus sp. SK3, Pseudomonas sp. ADMK77, Proteus sp. BLKB2 and Enterobacter hormaechei strain CW-3 by 16S rRNA gene sequencing. Phylogenetc analysis indicated the strains belong to three various genera namely, Proteus, Pseudomonas and Enterobacter. The evolutionary timeline of the bacteria was studied by constructing phylogenetic trees by Neighborhood Joining method. The presence of ESBL gene and biofilm forming capability were studied for the four strains. Antibiotic susceptibility patterns of the isolates were studied toward the commonly recommended antibiotics. Both the Proteus strains were found commonly susceptible to aminoglycoside and sulphonamide groups. Pseudomonas strain was found to be susceptible to cephems, aminoglycosides and fluoroquinolones. Enterobacter sp was found to be resistant to almost all antibiotic groups and susceptible to only sulphonamides group. The antibiotic susceptibility patterns of the bacteria help in choosing the empirical antibiotic treatment for UTI.

Chlorpyrifos-induced changes in the antioxidants and fatty acid compositions of Chroococcus turgidus NTMS12.

UNLABELLED: This study investigated the effect of pesticide chlorpyrifos (CP) on a freshwater cyanobacterium Chroococcus turgidus NTMS12. The changes in chlorophyll-a, proline, superoxide dismutase (SOD), catalase (CAT) activities and fatty acid composition of the test organism were analysed. Organism was grown at 6, 9 and 12 mg l(-1) of CP, and based on the chlorophyll-a content, 6 mg l(-1) of CP was found to be the tolerable concentration. Hence, 6 mg l(-1) of CP was taken to evaluate the concentration of proline and activities of SOD and CAT at 48-h exposure. The changes in the fatty acid profile were analysed after 7 days of exposure. Upon pesticide exposure, increased concentration of proline and activities of SOD and CAT were found. Significant changes in fatty acid profile have also been observed. However, polyunsaturated fatty acid content was decreased in treated cultures when compared with the untreated control. Changes in biochemical activities indicate that cyanobacteria C. turgidus NTMS12 undergo adaptive changes against CP-induced oxidative stress. SIGNIFICANCE AND IMPACT OF THE STUDY: : Chlorpyrifos induces oxidative stress in Chroococcus turgidus NTMS12. A strong inference was made on increased activity of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT) and proline content and lowering the level of unsaturated fatty acids under the pesticide-exposed condition. These significant changes are the defence mechanisms against the oxidative stress. Thus, this organism holds great promise in resisting toxic pesticide.

Role of gamma-oryzanol in drought-tolerant and susceptible cultivars of rice (Oryza sativa L.).

Drought-tolerant cultivars and their phytochemical composition, which has a role in providing drought tolerance are gaining importance. In this study, rice bran oil and semi-purified oryzanol (SPO) obtained from five rice (Oryza sativa L.) cultivars, namely P1401 and PB1 (drought-susceptible) and N22, PNR381 and APO (drought-tolerant) were analyzed for the gamma-oryzanol content, an antioxidant present in considerable amount in the rice bran. The higher level of gamma-oryzanol and its antioxidant activity was observed in drought-tolerant cultivars (N22, PNR381 and APO) as compared to drought-susceptible (PB1 and P1401), suggesting the role of gamma-oryzanol in drought tolerance, as antioxidants are known to play an important role by scavenging free radicals. The total antioxidant activity of gamma-oryzanol might be attributed to 24-methylene cycloartanyl ferulate, a major component of gamma-oryzanol. By enhancing the level of active oryzanol components identified in this study by genetic and molecular means could impart increased drought tolerance.

Socio-economic impact assessment of large-scale recycling of treated municipal wastewater for indirect groundwater recharge.

Reusing treated wastewater is an emerging solution to address freshwater scarcity, and surface water contamination faced worldwide. A unique large-scale wastewater recycling project was implemented to replenish groundwater by filling secondary treated wastewater (STW) into existing irrigation tanks in severely drought-hit areas of the Kolar districts of Southern India. This study quantifies the socio-economic impacts of this large-scale indirect groundwater recharge scheme. The changes in areas receiving STW i.e., impacted areas and those areas which did not receive STW i.e., non-impacted areas was studied. Also, pre and post recycling changes were quantified in the Kolar district. The results show that surface water quality meets India's most stringent treated wastewater discharge standards prescribed by the Hon'ble National Green Tribunal. Due to these recycling efforts, significant improvements in groundwater level and quality were found. It was observed that there was a noticeable difference in agricultural cropping areas, seasons, patterns, and production between impacted and non-impacted areas. Post-recycling, farmers tended to cultivate cash and water-intensive crops over less water-intensive crops. During the post-recycling period, livestock and milk production also increased, and in impacted areas, it was significantly higher. Post-recycling, fish production increased and land prices per hectare increased by 118 % in impacted areas. The farmer's net income under flowers and vegetable farming increased by 202 % and 150 % respectively in impacted areas compared to non-impacted areas. Furthermore, this project contributes to a circular economy transition in the water sector, which has economic, environmental, social, and cultural benefits. A key recommendation from the outcomes of the study is to draft and implement a policy that encourages the reuse of recycled water for groundwater recharge which in turn will improve the agro-economic system and food security.

Investigating the effects of irrigation with indirectly recharged groundwater using recycled water on soil and crops in semi-arid areas.

The utilization of direct wastewater for irrigation poses many environmental problems such as soil quality deterioration due to the accumulation of salts, heavy metals, micro-pollutants, and health risks due to undesirable microorganisms. This hampers its agricultural reuse in arid and semi-arid regions. To address these concerns, the present study introduces a recent approach that involves using indirectly recharged groundwater (GW) with secondary treated municipal wastewater (STW) for irrigation through a Soil Aquifer Treatment-based system (SAT). This method aims to mitigate freshwater scarcity in semi-arid regions. The study assessed GW levels, physicochemical properties, and microbial diversity of GW, and soil in both impacted (receiving recycled water) and non-impacted (not receiving recycled water) areas, before recycling (2015-2018) and after recycling (2019-2022) period of the project. The results indicated a significant increase of 68-70% in GW levels of the studied boreholes in the impacted areas. Additionally, the quality of indirectly recharged GW in the impacted areas improved notably in terms of electrical conductivity (EC), hardness, total dissolved solids (TDS), sodium adsorption ratio (SAR), along with certain cations and anions (hard water to soft water). No significant difference was observed in soil properties and microbial diversity of the impacted areas, except for EC and SAR, which were reduced by 50% and 39%, respectively, after the project commenced. The study also monitored specific microbial species, including total coliforms, Escherichia coli (as indicator organisms), Shigella, and Klebsiella in some of the harvested crops (beetroot, tomato, and spinach). However, none of the analysed crops exhibited the presence of the studied microorganisms. Overall, the study concludes that indirectly recharged GW using STW is a better sustainable and safe irrigation alternative compared to direct wastewater use or extracted hard GW from deep aquifers.

Added value of the measles-rubella supplementary immunization activity in reaching unvaccinated and under-vaccinated children, a cross-sectional study in five Indian districts, 2018-20.

INTRODUCTION: Supplementary immunization activities (SIAs) aim to interrupt measles transmission by reaching susceptible children, including children who have not received the recommended two routine doses of MCV before the SIA. However, both strategies may miss the same children if vaccine doses are highly correlated. How well SIAs reach children missed by routine immunization is a key metric in assessing the added value of SIAs. METHODS: Children aged 9 months to younger than 5 years were enrolled in cross-sectional household serosurveys conducted in five districts in India following the 2017-2019 measles-rubella (MR) SIA. History of measles containing vaccine (MCV) through routine services or SIA was obtained from documents and verbal recall. Receipt of a first or second MCV dose during the SIA was categorized as "added value" of the SIA in reaching un- and under-vaccinated children. RESULTS: A total of 1,675 children were enrolled in these post-SIA surveys. The percentage of children receiving a 1st or 2nd dose through the SIA ranged from 12.8% in Thiruvananthapuram District to 48.6% in Dibrugarh District. Although the number of zero-dose children prior to the SIA was small in most sites, the proportion reached by the SIA ranged from 45.8% in Thiruvananthapuram District to 94.9% in Dibrugarh District. Fewer than 7% of children remained measles zero-dose after the MR SIA (range: 1.1-6.4%) compared to up to 28% before the SIA (range: 7.3-28.1%). DISCUSSION: We demonstrated the MR SIA provided considerable added value in terms of measles vaccination coverage, although there was variability across districts due to differences in routine and SIA coverage, and which children were reached by the SIA. Metrics evaluating the added value of an SIA can help to inform the design of vaccination strategies to better reach zero-dose or undervaccinated children.

Hemostatic interference of Indian king cobra (Ophiophagus hannah) Venom. Comparison with three other snake venoms of the subcontinent.

Unlike Naja naja, Bungarus caeruleus, Echis carinatus, and Daboia/Vipera russellii venoms, Ophiophagus hannah venom is medically ignored in the Indian subcontinent. Being the biggest poisonous snake, O. hannah has been presumed to inject several lethal doses of venom in a single bite. Lack of therapeutic antivenom to O. hannah bite in India makes any attempt to save the victim a difficult exercise. This study was initiated to compare O. hannah venom with the above said venoms for possible interference in hemostasis. Ophiophagus hannah venom was found to actively interfere in hemostatic stages such as fibrin clot formation, platelet activation/aggregation, and fibrin clot dissolution. It decreased partial thromboplastin time (aPTT), prothrombin time (PT), and thrombin clotting time (TCT). These activities are similar to that shown by E. carinatus and D. russellii venoms, and thus O. hannah venom was found to exert procoagulant activity through the common pathway of blood coagulation, while N. naja venom increased aPTT and TCT but not PT, and hence it was found to exert anticoagulant activity through the intrinsic pathway. Venoms of O. hannah, E. carinatus, and D. russellii lack plasminogen activation property as they do not hydrolyze azocasein, while they all show plasmin-like activity by degrading the fibrin clot. Although N. naja venom did not degrade azocasein, unlike other venoms, it showed feeble plasmin-like activity on fibrin clot. Venom of E. carinatus induced clotting of human platelet rich plasma (PRP), while the other three venoms interfered in agonist-induced platelet aggregation in PRP. Venom of O. hannah least inhibited the ADP induced platelet aggregation as compared to D. russellii and N. naja venoms. All these three venoms showed complete inhibition of epinephrine-induced aggregation at varied doses. However, O. hannah venom was unique in inhibiting thrombin induced aggregation.

Reduction and fixation of cranial cervical fracture/luxations using screws and polymethylmethacrylate (PMMA) cement: a distraction technique applied to the base of the skull in thirteen dogs.

OBJECTIVE: To (1) describe a surgical distraction technique for C1-2 cervical fractures/luxations or atlantoaxial (AA) subluxations using the base of the skull (basion of the foramen magnum) and either C2-3 or C3-4 for the purchase points of intraoperative axial distraction and (2) report outcome in 13 dogs. STUDY DESIGN: Retrospective case series. ANIMALS: Dogs (n = 13). MATERIALS AND METHODS: Medical records (September 1995-December 2005) of dogs with fracture/luxation of the cervical spine, or AA subluxations, were reviewed. Only dogs that had intraoperative linear distraction using the base of the skull as a purchase point for a self-retaining retractor were included. Signalment, presenting neurologic deficits, fracture location, and concurrent injury were recorded. Both short-term in-hospital follow-up, including healing and any complications, and long-term telephone follow-up were obtained. RESULTS: Realignment of the spinal vertebrae, reestablishing the normal properties of the spinal canal, was achieved after distraction in all dogs. Screws and small pins incorporated into polymethylmethacrylate cement were used to span the fracture ventrally and achieve rigid internal fixation. Eight dogs had a complete neurologic recovery, 2 dogs had slight residual ataxia, 2 dogs died, and 1 dog was lost to follow-up. Healing was good (mean, 7.5 weeks) or excellent (mean, 5.1 months) based on in-hospital follow-up (mean, 5.1 months). On final telephone follow-up (mean, 7.7 years), no dogs were reported to have had any associated problems or additional surgery. CONCLUSIONS: This surgical distraction technique was a reliable, relatively simple method to obtain reduction of fracture/luxations of C1-2 to re-align the spinal canal. Mortality in this series appears lower than that previously reported and supports surgical management of these injuries.

Evaluation of orbital volume in unilateral orbital fracture using computed tomography.

Aim and Objective: Our study was an observational and evolutional to analyze the significance of orbital volume calculation in predicting the probability or tendency of developing late enopthalmous on patients with unilateral orbital fracture with or without associated midface injury reported to our hospital. Materials and Methods: The subjects in our study included fifteen patients between18 and 60years. They were treated surgically and conservatively. Patients were exposed to series of examination from the day of injury or reporting for a consecutive period of up to six months following injury. CT was taken prior to treatment following management to evaluate the volume change in orbit. Its correlation and significance to treatment outcome were analyzed using MIMICS software by Materialise by comparing injured orbit to opposite side normal eye. Result: Conclusive of clinical examination and summary of statistical analysis patients were categorized into three groups: Low (with volume change of less than 2%), Medium (volume change between 2 to 20%) and High (volume change more than 20%) by comparing fractured orbit to uninjured orbit and a statistically significant mean value of 27.7 ± 2.6 cu. mm for fractures eye and 25.9 ± 2.6 cu. mm for normal eye was found (P ≤ 0.05). Conclusion: Orbital volume can be considered as useful criteria in evaluation of patients with orbital fracture. Evaluation and follow up in correlation with resultant volume assessment provide a better indicator for assessment of prognosis and enopthalmous. Unlike other methods for volume assessment MIMICS software by Materialise provide a better method and it can be incorporated to working station in hospital. Every CT assessment in orbital trauma should include orbital volume as a parameter and this software is easy and user-friendly software that does not require its technical knowledge for application.

The metalloprotease, NN-PF3 from Naja naja venom inhibits platelet aggregation primarily by affecting α2ß1 integrin.

NN-PF3 is a non-toxic, anticoagulant, high-molecular-mass (67.81 kDa) metalloprotease from Indian cobra (Naja naja) venom. In the present study, NN-PF3 was investigated for the mechanism of inhibition of collagen-induced aggregation of human platelets. The complete inhibition of collagen-induced aggregation and partial inhibition of ADP- and epinephrine-induced aggregation has the respective IC(50) of 75 ± 5, 185 ± 10, and 232 ± 12 nM, whereas no inhibition of thrombin-, arachidonic acid-, and ristocetin-induced aggregation of platelets was observed in platelet-rich plasma. Further, native NN-PF3 and EDTA-inactivated NN-PF3 inhibited collagen-induced aggregation of washed platelets with respective IC(50) of 75 ± 4 and 180 ± 6 nM. The higher inhibitory effect of native NN-PF3 compared with EDTA-inactivated NN-PF3 suggests the enzymatic and non-enzymatic mechanism of inhibition. NN-PF3 pretreatment affected the collagen binding but not the fibrinogen, and fibronectin binding of washed platelets in adhesion assay suggested that the collagen receptors are affected. Western blot study using anti-integrin α2ß1 mAb 6F1 suggested that NN-PF3 binds to integrin α2ß1 in a primary structure-dependent manner only and is not cleaved. There was a drastic reduction in the intensity of several intracellular signaling phosphotyrosine protein bands when monoclonal anti-phosphotyrosine antibody was used, suggesting that the major activation pathway of platelets get affected, which occurs through glycoprotein VI. NN-PF3 did not bind to collagen as revealed by Western blot using anti-collagen mAb. Furthermore, neither the proteolytic cleavage of fibrinogen nor its degradation products by NN-PF3 contributed for the collagen-induced platelet aggregation inhibition.

Subcritical water extraction of antioxidant compounds from Seabuckthorn (Hippophae rhamnoides) leaves for the comparative evaluation of antioxidant activity.

A novel environmentally friendly technique, subcritical water extraction (SWE) was employed for the extraction of antioxidant compounds from Seabuckthorn leaves (SBT). Antioxidant activity of the extracts was evaluated using commonly accepted chemical assays. Also, present study reports the cytoprotective and antioxidant properties of SBT against tertiary-butyl hydroperoxide (tert-BOOH) induced oxidative stress in murine macrophages (Raw 264.7). Exposure of cells to tert-BOOH resulted, increase in cytotoxicity, reactive oxygen species (ROS) production and decrease in mitochondrial membrane potential, which is responsible for fall in intracellular antioxidant levels. Pretreatment of cells with SBT extracts inhibited cytotoxicity, ROS production and maintained antioxidants levels similar to that of control cells. The chemical composition of the SWE extracts studied showed total phenol content (76.07-93.72mg/g GAE) and total flavonoid content (47.06-66.03mg/g rutin). Further, some of its phenolic constituents; (1) Quercetin-3-galactoside, (2) Kaempferol and (3) Isorhamnetin were quantified by RP-HPLC.

To Study the Role of Perfusion Index as a Predictor of Hypotension during Spinal Anesthesia in Lower Segment Cesarean Section - A Prospective Observational Study.

Background and Aims: Spinal anesthesia has become the choice of neuraxial anesthesia for cesarean delivery. Hypotension, the most common side effect of spinal anesthesia is more in parturients. Perfusion index (PI) is one of the newest methods, used noninvasively for the prediction of hypotension postspinal anesthesia, evaluation of regional block success, and a tool for pain assessment. In our study, we have used PI for predicting the occurrence of hypotension postsubarachnoid block in elective lower segment cesarean section (LSCS). Methods: In this prospective observational study, parturients posted for elective LSCS under spinal anesthesia were included. Spinal anesthesia was performed at L3-L4 interspace using 25G (gauge) Quincke needle, with 1.8 mL of 0.5% hyperbaric bupivacaine plus 20 µg fentanyl. Hypotension was defined as a decrease in systolic blood pressure >25% from the baseline. Statistical Analysis Used: Statistical analysis was performed using the paired sample t-test, Chi-square test, and independent t-test. Results: From our study, we had found that a positive correlation exists between the baseline PI of the patient and the occurrence of hypotension intraoperatively. A cutoff value of 1.75 (with P < 0.001) was found to be significant; above which the occurrence of hypotension was definite. The sensitivity of this cutoff value was 75% and specificity was 71% for this value. Conclusion: Baseline perfusion of >1.75 can predict hypotension postspinal anesthesia in LSCS. The incidence of hypotension was about 93% in parturients whose baseline was above the cutoff value.

Anti-coagulant activity of a metalloprotease: further characterization from the Indian cobra (Naja naja) venom.

A high molecular mass, non toxic metalloprotease the NN-PF3 with the bound Ca(2+) and Zn(2+) from the Naja naja venom has been studied further for its anticoagulant property. The molecular mass by MALDI-TOF mass spectrometry was 67.81 kDa. The NN-PF3 exhibited fibrin(ogen)olytic activity. In addition to fibrinogen, NN-PF3 hydrolyzed blood and plasma clot with the later hydrolyzed about one fold higher. The alpha polymer of fibrin was preferentially hydrolyzed over the alpha chain but the beta chain and gamma-gamma dimer remained untouched. It was devoid of plasminogen activation property. It prolonged the activated partial thromboplastin time, prothrombin time and the thrombin clotting time of citrated human plasma. It did not affect the thrombin activity. In mice, defibrinogentaion, prolonged bleeding time (P < 0.01) and reduced fibrinogen level were observed following intravenous injection. Human plasma or alpha2-macroglobulin did not, but the polyvalent anti-venom inhibited the NN-PF3 activity. In contrast to most snake venom metalloproteases, it did not degrade extra cellular matrix proteins.