The initial experience of COVID-19 vaccination from a tertiary care centre of India.

To the Editor Drugs Controller General of India (DCGI) has approved the Bharat biotech vaccine against COVID-19, which is a locally manufactured inactivated vaccine named ''COVAXIN'' in collaboration with the Indian council of medical research (ICMR) on 3rd  January 2021 for emergency use along with the "Covishield" Oxford-AstraZeneca vaccine manufactured locally by the Serum Institute of India... *The Safdarjung COVID-19 Vaccination group: KR Meena (Professor, Paediatrics), U Venkatesh (Assistant Professor, Community Medicine), Pushpa Kumari (Associate Professor, Medicine), Sonal Burman (Specialist, Medicine), Neeraj Kumar Gupta (Professor and Head, Pulmonary Medicine), Nitesh Gupta (Assistant Professor, Pulmonary Medicine), Rohit Kumar (Assistant Professor, Pulmonary Medicine), Swetabh Purohit (Senior resident, Pulmonary Medicine), Arjun Ramaswamy (Senior resident, Pulmonary Medicine).

Adsorptive removal of lead (Pb), copper (Cu), nickel (Ni) and mercury (Hg) ions from water using chitosan silica gel composite.

Silica gel chitosan composite was prepared to perform adsorptive experiment of different heavy metal ion solutions. The characterization of chitosan + silica gel (Ch + Sg) composite was done by FTIR and SEM-EDS to understand the presence of active sites and to have an insight on the surface morphology. The adsorption study of heavy metal ions by Ch + Sg composite gives maximum removal percent for Cu, Pb and Ni which were obtained at pH 5 and for Hg at pH 6.The trend of removal by Ch + Sg signifies that maximum removal percent was attained at 120 min. The surface of Ch + Sg is heterogeneous for the adsorption of Hg, Ni and Cu and homogeneous for Pb adsorption. The values obtained for Pb signify that its adsorption best fitted to pseudo first order with the R2 value of 0.986, whereas pseudo second order best fitted to the experimental data of Cu, Ni and Hg as R2 values which are 0.983, 0.819 and 0.957 respectively. The values of change in entropy (⊿S) obtained for Pb, Cu, Ni and Hg are - 69.33, - 118, - 63.33 and - 98.52 J/mol K respectively. Negative values of change in enthalpy, ⊿H in (kJ/mol) are in the range of - 18.2 to - 37.66 which indicates both physical and chemical adsorption involves in the process of adsorption.

Characterization and pollutant removal efficiency of biochar derived from baggase, bamboo and tyre.

Conversion of broad-spectrum organic waste into carbonaceous biochar has gained enormous interest in past few years. The present study aims to characterize feedstock (FS), i.e. bagasse (Bg), bamboo (Bm) and biochar (BC), i.e. baggase biochar (BBg), bamboo biochar (BBm) and tyre biochar (Ty). Significant changes in elemental composition, atomic ratio, proximate analyses, mineral content and heavy metal content were observed which was well supported by Fourier transform infrared (FTIR) spectroscopy and X-ray diffraction (XRD) analysis. Impregnation with ferric hydroxide was done, and resultant modified biochars (MBC), i.e. iron-impregnated baggase biochar (FeBBg), iron-impregnated bamboo biochar (FeBBm) and iron-impregnated tyre biochar (FeTy), along feedstock and biochar were used for PO4 (3-), Pb, Hg and Cu adsorption. In general, BBg, FeBBg, BBm, FeBBm, Ty and FeTy were found to adsorb PO4 (3-), Pb, Hg and Cu better than Bg and Bm, except in few cases. Results from adsorption experiments were fitted into Langmuir, Freundlich and Temkin models of isotherms and pseudo-first-order, pseudo-second-order and Elovich models of kinetics. Result of batch study adsorption revealed that maximum adsorption of PO4 (3-), Pb, Hg and Cu was done by FeBBg (adsorption mechanism explained by Freundlich model), FeTy (Temkin model), Ty (Langmuir model) and BBm (Langmuir model) respectively. According to R (2) values, pseudo-first-order reaction was well suited to PO4 (3-), Pb, Hg and Cu adsorption. The optimum pH for maximum adsorption was observed to be 7.4 for PO4 (3-), 5 for Cu and 6 for Pb and Hg respectively.

Bcl-2-mediated drug resistance: inhibition of apoptosis by blocking nuclear factor of activated T lymphocytes (NFAT)-induced Fas ligand transcription.

Bcl-2 inhibits apoptosis induced by a variety of stimuli, including chemotherapy drugs and glucocorticoids. It is generally accepted that Bcl-2 exerts its antiapoptotic effects mainly by dimerizing with proapoptotic members of the Bcl-2 family such as Bax and Bad. However, the mechanism of the antiapoptotic effects is unclear. Paclitaxel and other drugs that disturb microtubule dynamics kill cells in a Fas/Fas ligand (FasL)-dependent manner; antibody to FasL inhibits paclitaxel-induced apoptosis. We have found that Bcl-2 overexpression leads to the prevention of chemotherapy (paclitaxel)-induced expression of FasL and blocks paclitaxel-induced apoptosis. The mechanism of this effect is that Bcl-2 prevents the nuclear translocation of NFAT (nuclear factor of activated T lymphocytes, a transcription factor activated by microtubule damage) by binding and sequestering calcineurin, a calcium-dependent phosphatase that must dephosphorylate NFAT to move to the nucleus. Without NFAT nuclear translocation, the FasL gene is not transcribed. Thus, it appears that paclitaxel and other drugs that disturb microtubule function kill cells at least in part through the induction of FasL. Furthermore, Bcl-2 antagonizes drug-induced apoptosis by inhibiting calcineurin activation, blocking NFAT nuclear translocation, and preventing FasL expression. The effects of Bcl-2 can be overcome, at least partially, through phosphorylation of Bcl-2. Phosphorylated Bcl-2 cannot bind calcineurin, and NFAT activation, FasL expression, and apoptosis can occur after Bcl-2 phosphorylation.

Quantification of nitrogen transformation and leaching response to agronomic management for maize crop under rainfed and irrigated condition.

Nitrogen (N) plays an important role in agriculture crop production but the increasing application of nitrogen increases the possibilities of groundwater contamination through nitrate leaching. Nitrate leaching is the inevitable part of agriculture production which occurs during nitrogen fertilization. Hence, the quantification of nitrogen fertilizer is required to reduce nitrate leaching. In this study, nitrogen transformation and transport such as ammonium (NH4+) and nitrate (NO3-) at different soil depths and maize crop growth stages were measured during field experiments for two sowing dates (timely and delay) and four N fertilization levels under irrigated (year 2013 and 2014) and rainfed (year 2012 and 2014) conditions for maize crop. NH4+, NO3- and total nitrogen concentrations were measured using spectrophotometer at 410 nm and Kjeldahl method at varying soil depths and maize crop growth stages. Thereafter, nitrogen balance approach was used to estimate the NO3- leaching. Results indicated that NO3- leaching in irrigated condition was higher 109% in N75, 179% in N100, and 292% in N125 level respectively in comparison to the N0 level in timely sowing date, while in delayed sowing date, leaching was higher 54% in N75, 123% in N100, and 184% in N125 level respectively in comparison to N0 level. In rainfed, the NO3- leaching was higher 30% in N60, 59% in N80, and 99% in N100 level respectively in comparison to N0 level for the timely sowing date, while in delayed sowing, leaching was higher 23% in N60, 44% in N80, and 78% in N100 level respectively in comparison to N0 level. The results indicate that leaching losses were less in timely sowing dates for both rainfed and irrigated maize. The study further reveals that sowing dates combination with N levels could be an effective management strategy to reduce NO3- leaching by minimizing the N fertilization.

Galapagos hydrothermal mounds: stratigraphy and chemistry revealed by deep-sea drilling.

The Galápagos mounds sea-floor hydrothermal system is at least 300,000 years old and once produced manganese-poor sediments, which nearly blanketed the area of the present mounds field. Present-day mound deposits are limited manganese-rich exposures, suggesting that the system has changed from rock-to water-dominated and has diminished in intensity with time.

Chronic kidney disease in India: challenges and solutions.

Chronic diseases have become a major cause of global morbidity and mortality even in developing countries. The burden of chronic kidney disease (CKD) in India cannot be assessed accurately. The approximate prevalence of CKD is 800 per million population (pmp), and the incidence of end-stage renal disease (ESRD) is 150-200 pmp. The most common cause of CKD in population-based studies is diabetic nephropathy. India currently has 820+ nephrologists, 710+ hemodialysis units with 2,500+ dialysis stations and 4,800+ patients on CAPD. There are 172+ transplant centers, two-thirds of which are in South India and mostly privately run. Nearly 3,500 transplants are done annually, the total number of cadaver donors being approximately 700 till now. Thus, taken together, nearly 18,000-20,000 patients (10% of new ESRD cases) in India get renal replacement therapy. The cost of single hemodialysis varies between USD 15 and 40 with an additional cost of erythropoietin being USD 150-200/month. The cost of CAPD using a 'Y' set with 3 exchanges/week is USD 400/month. The cost of the transplant procedure in a state-run hospital is USD 800-1,000, and the cost of immunosuppression using tacrolimus, steroid and mycophenolate is USD 350-400/month. Until recently, the government did not recognize CKD/ESRD as a significant problem in India. However, some illustrious activities in relation to CKD brought attention of the media and policymakers to this very common but till now deprived group of diseases. On the one side the government has initiated a process by which it is planning to establish stand-alone hemodialysis units in the country to increase the facilities at an affordable cost, and on the transplant side it had launched a National Organ Transplant Program to facilitate transplantation on a national scale. Hemodialysis program is halfway to being implemented. Thus, in India there is still a long way to go with respect to CKD. Until then, in a country like India, screening of high-risk individuals for CKD and the risk factors is the best bet.

In vitro effect of fluoroquinolones against Mycobacterium tuberculosis isolates from Agra & Kanpur region of north India.

BACKGROUND & OBJECTIVE: Fluoroquinolones (FQs) are important drugs used for treatment of drug resistant tuberculosis and are also now being considered as first line drugs to shorten the duration of treatment of tuberculosis (TB). In order to find out useful FQs for treatment of tuberculosis, the comparative efficacy of five FQs, namely, ofloxacin (OFL), ciprofloxacin (CIP), sparfloxacin (SPX), gatifloxacin (GAT) and levofloxacin (LEVX) was studied against Mycobacterium tuberculosis (MTB) isolates obtained from both treated and untreated patients from Agra and Kanpur regions of north India. METHODS: A total of 162 MTB isolates [including 110 MTB isolates obtained from untreated patients (Cat-I) and 52 isolates from treated patients (Cat-II)] were tested for their susceptibilities to FQs using standard minimum inhibitory concentration (MIC) method on Löwenstein-Jensen medium. RESULTS: Keeping in view the therapeutically achievable drug levels, it was found that in Cat-I 97.2 per cent (107/110) isolates were sensitive to GAT, 89 per cent (98/110) to LEVX at 1 microg/ml whereas 92.7 per cent (102/110) isolates were inhibited by OFL at 2 microg/ml and 73.6 per cent (81/110) to SPX at 0.5 microg/ml. Only 63.6 per cent (70/110) isolates were found to be sensitive to CIP at 2 microg/ml which increased to 89 per cent (98/110) at 4 microg/ml (higher than achievable peak serum level). On the other hand, among 52 isolates for Cat-II, 37 (71.2%) were found to be sensitive to GAT and 33 (63.5%) to LEVX at 1 microg/ml concentration, 28 (53.8%) to SPX at 0.5 microg/ml whereas 33 (63.5%) and 24 (46.2%) isolates were found to be sensitive to OFL and CIP at 2 microg/ml, respectively. INTERPRETATION & CONCLUSION: It appears that GAT has higher activity against MTB isolates followed by OFL, LEVX and SPX whereas CIP showed the lowest activity. GAT was also found to be the most effective FQ against multi-drug resistant (MDR) isolates both from Cat-I and Cat-II patients. Thus, except CIP, other FQs showed potential to be included in the treatment regimens of tuberculosis including MDR-TB.

Treatment of rayon grade pulp drain effluent by upflow anaerobic fixed packed bed reactor (UAFPBR).

The Rayon grade pulp (RGP) drain effluent of pulp and paper mill was studied to find out pollutant loading and its control measures by low cost and efficient treatment method. Upflow anaerobic fixed packed bed reactor (UAFPBR) with brick ballasts as packing material was used for this purpose. This was compared with conventional anaerobic treatment method. The digested slurry was taken as inoculum from the active cow dung biogas plant. After stabilization of the reactors the reduction in pollutant loading was found to be higher in UAFPBR than conventional anaerobic reactor (CAR). Hydraulic retention time (HRT) of 12 hr was optimum for the treatment of effluent when 74.5% COD and 81% BOD reduction was obtained. 30% inoculum concentration was best for the anaerobic treatment of RGP colour drain effluent. The maximum biogas production (1.37 l l(-1) of effluent) was when the effluent was inoculated with 30% seeding material. Thus, UAFPBR system was very efficient in terms of BOD, COD, TSS and TDS removal from RGP drain of paper mills in ambient environmental conditions.

Assessment of utilization of RCH services and client satisfaction at different level of health facilities in Varanasi District.

OBJECTIVE: To assess the various factors influencing utilization and non-utilization of RCH services and extent of client satisfaction. METHODS: A cross-sectional study was conducted during October to December 2008 at two selected blocks of Varanasi district, Uttar Pradesh. Principal study subjects were 509 women having children less than 12 months old, selected through a multistage sampling technique. Data were collected through in-depth interview and Focus Group Discussions conducted among the beneficiaries of the services. RESULTS: The study revealed that utilisation of the RCH services in the government facilities was higher among the backward classes than the general category; higher the level of education the lower was the utilisation of government services. Over all, 16% of the respondents were not satisfied with government facilities. 25% of the SC category was not satisfied with the services in spite of being the main users. Among RCH services utilization was highest (89%) for antenatal care services (ANC). 41.6% respondents did not receive any Post Natal Care (PNC) after their most recent birth. About 30% deliveries were at home out of which only 10% received PNC whereas out of 70% institutional deliveries about 80% received PNC. Overall 16.3 % of the respondents were not satisfied with the services provided by government health facilities. Around 16% and 14% were not satisfied with the behavior of medical officer and the health workers respectively and non-satisfaction was highest among SC category. CONCLUSION: All health facilities need to be made functional according to Indian Public Health Standards (IPHS) of NRHM.

Experience with various reconstructive techniques for meningomyelocele defect closure in India.

BACKGROUND: The estimated incidence of spina bifida is 1-2 cases per 1000 population. In earlier literature, the global prevalence of meningomyelocele (MMC) is reported as 0.8-1.0 per 1000 live births. This retrospective study analyses the outcome of various surgical procedures performed for the closure of MMC defects. METHOD: A total of 22 patients with MMC defects who underwent repair at our institute from July 2016 to August 2018 were included in the study. A retrospective review of all the cases operated was completed to analyse patient demography including defect size, defect location, surgical procedures, complications and the final outcome. RESULTS: Out of 22 cases, wherein the neurosurgery department sought help from the plastic surgery department, 11 defects were closed using the Limberg flap technique, 4 defects were closed with either primary closure or the double flap rotation flaps, one defect was closed using the triple rotation flap and 2 defects were closed using the local transposition flap cover technique. Complications were noted in only three cases. One patient had a local wound infection, while in two other cases, wound dehiscence was observed. All 3 cases were managed conservatively. On average, it takes approximately 70 days in India to close such defects. CONCLUSION: MMC defects can be effectively managed with local flap options such as Limberg flap, local transposition flap or rotation flaps. Various reasons for the delay in closure were reported in patients late to our centre, when the first point of contact was with other departments.

Influence of diabetes on liver injury induced by antitubercular drugs and on silymarin hepatoprotection in rats.

Isoniazid, rifampicin and pyrazinamide during short-course chemotherapy for tuberculosis can result in liver injury. The coexistence of tuberculosis and diabetes is common in patients who receive inadequate treatment. The risk of hepatotoxicity from many toxicants is increased in diabetic rats. Silymarin provides protection against liver injury caused by many hepatotoxicants, including antitubercular drugs (ATDs). In the wake of increased severity of ATD-induced hepatotoxicity in diabetes we report here the results of a study on the influence of diabetes on silymarin hepatoprotection in rats. Rats with diabetes induced via intraperitoneally injected streptozotocin (50 mg/kg), nondiabetic rats and insulin-treated diabetic rats received isoniazid (7.5 mg/kg/day), rifampicin (10 mg/kg/day) and pyrazinamide (35 mg/kg/day) orally (p.o.) with or without silymarin (100 mg/kg/day p.o.) treatment for 45 days. Compared to nondiabetic rats, liver function tests and histological changes of liver revealed exaggerated liver injury in diabetic rats caused by ATDs which was evident by 5- to 8-fold increases in serum levels of marker enzymes (aspartate and alanine aminotransferase, alkaline phosphatase and gamma-glutamyltranspeptidase) and 1- to 2-fold increases in bilirubin accompanied by a 2-fold decrease in total serum proteins, intense fatty and inflammatory infiltrations, necrosis and fibrosis. Coadministration of silymarin provided protection against ATD hepatotoxicity in all animals. However, insulin-treated diabetic animals showed greater silymarin-induced hepatoprotection against ATD-induced liver injury, which was characterized by near normal levels of marker enzymes, an increase in total proteins and normal hepatic structure. These results thus indicate that diabetes exaggerates ATD-induced liver injury and attenuates silymarin-induced hepatoprotection. However, insulin treatment for diabetes offers greater silymarin-induced hepatoprotection against ATD-induced liver injury.

A review on sustainable yeast biotechnological processes and applications.

Yeast is very well known eukaryotic organism for its remarkable biodiversity and extensive industrial applications. Saccharomyces cerevisiae is one of the most widely used microorganisms in biotechnology with successful applications in the biochemical production. Biological conversion with the focus on the different utilization of renewable feedstocks into fuels and chemicals has been intensively investigated due to increasing concerns on sustainability issues worldwide. Compared with its counterparts, Saccharomyces cerevisiae, the baker's yeast, is more industrially relevant due to known genetic and physiological background, the availability of a large collection of genetic tools, the compatibility of high-density and large-scale fermentation, and optimize the pathway for variety of products. Therefore, S. cerevisiae is one of the most popular cell factories and has been successfully used in the modern biotech industry to produce a wide variety of products such as ethanol, organic acids, amino acids, enzymes, and therapeutic proteins. This study explores how different sustainable solutions used to overcome various environmental effects on yeast. This work targets a broad matrix of current advances and future prospect in yeast biotechnology and discusses their application and potential in general.

Tuberculosis diagnostic and treatment practices in private sector: Implementation study in an Indian city.

SETTING: Implementation study in private health facilities in an Indian metropolis. OBJECTIVES: Improve Tuberculosis (TB) care by private practitioners (PPs). METHODS: PPs from a defined city area were imparted short training in TB care and linkages made with public facilities; subsequent practices were recorded. RESULTS: Of 364 presumptive TB patient records, 70 (19.3%) did not conform to its definition. Of the conforming, 174 (59.2%) had presumptive pulmonary TB (PTB), 53 (18%) presumptive extra-pulmonary (EPTB) and 67 (24%) had both. Of conforming presumptive PTB, most underwent Chest X-ray and sputum examination in private laboratories. Tissue based diagnostics were not advised for most presumptive EPTB patients. Of 101 cases diagnosed with TB, 82% were new, 23% known diabetic and 4.7% human immune deficiency virus (HIV) reactive out of 64 tested. Most were notified and initiated treatment within 15 days of diagnosis. One-fourth was prescribed standard treatment regimen and treatment was not directly observed for most. One third was initial defaulters or lost during treatment; 62% of PTB and 46% EPTB cases initiated on treatment in private were successfully treated. Of successfully treated PTB cases, 61% had undergone follow-up sputum examination. CONCLUSION: Much intensified support mechanisms are needed to improve TB care in private sector.

Knowledge of private practitioners of Bangalore city in diagnosis, treatment of pulmonary tuberculosis and compliance with case notification.

One hundred and twenty-nine qualified private practitioners (PPs) were assessed on their knowledge in diagnosis of pulmonary tuberculosis (PTB), treatment of a new drug sensitive PTB case and practices of case notification, using semi-structured questionnaire. About 20% had adequate knowledge of diagnosis, 29% of treatment regimen, 54% the need for Direct Observation Treatment and 57% about role of sputum smear examination in monitoring treatment response. Of 85 (68%) PPs who had diagnosed any TB case during last two years, 54 (64%) had practised notification. These findings suggest the need for upgrading knowledge of PPs in TB diagnosis, treatment and notification.

Bcl-2 and Bcl-X(L) block thapsigargin-induced nitric oxide generation, c-Jun NH(2)-terminal kinase activity, and apoptosis.

The proteins Bcl-2 and Bcl-X(L) prevent apoptosis, but their mechanism of action is unclear. We examined the role of Bcl-2 and Bcl-X(L) in the regulation of cytosolic Ca(2+), nitric oxide production (NO), c-Jun NH(2)-terminal kinase (JNK) activation, and apoptosis in Jurkat T cells. Thapsigargin (TG), an inhibitor of the endoplasmic reticulum-associated Ca(2+) ATPase, was used to disrupt Ca(2+) homeostasis. TG acutely elevated intracellular free Ca(2+) and mitochondrial Ca(2+) levels and induced NO production and apoptosis in Jurkat cells transfected with vector (JT/Neo). Buffering of this Ca(2+) response with 1, 2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetra(acetoxymethyl) ester (BAPTA-AM) or inhibiting NO synthase activity with N(G)-nitro-L-arginine methyl ester hydrochloride (L-NAME) blocked TG-induced NO production and apoptosis in JT/Neo cells. By contrast, while TG produced comparable early changes in the Ca(2+) level (i.e., within 3 h) in Jurkat cells overexpressing Bcl-2 and Bcl-X(L) (JT/Bcl-2 or JT/Bcl-X(L)), NO production, late (36-h) Ca(2+) accumulation, and apoptosis were dramatically reduced compared to those in JT/Neo cells. Exposure of JT/Bcl-2 and JT/Bcl-X(L) cells to the NO donor, S-nitroso-N-acetylpenacillamine (SNAP) resulted in apoptosis comparable to that seen in JT/Neo cells. TG also activated the JNK pathway, which was blocked by L-NAME. Transient expression of a dominant negative mutant SEK1 (Lys-->Arg), an upstream kinase of JNK, prevented both TG-induced JNK activation and apoptosis. A dominant negative c-Jun mutant also reduced TG-induced apoptosis. Overexpression of Bcl-2 or Bcl-X(L) inhibited TG-induced loss in mitochondrial membrane potential, release of cytochrome c, and activation of caspase-3 and JNK. Inhibition of caspase-3 activation blocked TG-induced JNK activation, suggesting that JNK activation occurred downstream of caspase-3. Thus, TG-induced Ca(2+) release leads to NO generation followed by mitochondrial changes including cytochrome c release and caspase-3 activation. Caspase-3 activation leads to activation of the JNK pathway and apoptosis. In summary, Ca(2+)-dependent activation of NO production mediates apoptosis after TG exposure in JT/Neo cells. JT/Bcl-2 and JT/Bcl-X(L) cells are susceptible to NO-mediated apoptosis, but Bcl-2 and Bcl-X(L) protect the cells against TG-induced apoptosis by negatively regulating Ca(2+)-sensitive NO synthase activity or expression.

Involvement of microtubules in the regulation of Bcl2 phosphorylation and apoptosis through cyclic AMP-dependent protein kinase.

The Bcl2 family of proteins plays a significant role in regulation of apoptosis. In this study, the microtubule-damaging drugs paclitaxel, vincristine, and vinblastine induced Bcl2 hyperphosphorylation and apoptosis in MCF-7 and MDA-MB-231 cells and reduced Bcl2-Bax dimerization. Paclitaxel or vincristine induced increased expression of Bax, while overexpression of Bcl2 in these cell lines counteracted the effects of low doses of these drugs. In addition, paclitaxel- and vincristine-induced activation of cyclic AMP (cAMP)-dependent protein kinase (protein kinase A [PKA]) induced Bcl2 hyperphosphorylation and apoptosis, which were blocked by the PKA inhibitor Rp diastereomers of cAMP (Rp-cAMP). This finding suggests that activation of PKA due to microtubule damage is an important event in Bcl2 hyperphosphorylation and induction of apoptosis. These microtubule-damaging drugs caused growth arrest in G2-M phase of the cell cycle and had no effect on p53 induction, suggesting that hyperphosphorylation mediated inactivation of Bcl2 and apoptosis without the involvement of p53. By comparison, the DNA-damaging drugs methotrexate and doxorubicin had no effect on Bcl2 hyperphosphorylation but induced p53 expression. Interestingly, paclitaxel or vincristine induced activation of caspase 3 and cleavage of poly(ADP-ribose) polymerase downstream of Bcl2 hyperphosphorylation. These data suggest that there may be a signaling cascade induced by agents that disrupt or damage the cytoskeleton that is distinct from (i.e., p53 independent), but perhaps related to (i.e., involves kinase activation and leads to apoptosis), the cellular response to DNA damage.

Application of Doehlert design in optimizing the determination of degraded products of nerve agents by ion-pair liquid chromatography electrospray ionization tandem mass spectrometry.

A qualitative method was developed for the determination of degraded products of nerve agents by using ion-pair liquid chromatography electrospray ionization tandem mass spectrometry (IP-LC-ESI-MS(n)). Generally, alkylphosphonic acids (APAs) and O-alkyl alkylphosphonic acids (AAPAs) give deprotonated molecular ion [M-H](-) in negative mode. Interestingly, first time we obtained the molecular radical anion [M](.-) of phosphonic acids in negative mode by using tri-n-butyl amine as an ion-pairing agent. We interpreted this observation as an indication of electrochemical reduction of phosphonic acids in electrospray needle. Three variables such as sheath gas flow, electrospray needle voltage and pH of the mobile phase were investigated to enhance the molecular radical anion [M](.-) signal of each analyte. The Doehlert design was used to obtain the region in which the optimum value of such variables is simultaneously achieved. Limit of detection achieved was 0.5 microg mL(-1) for AAPAs and 10 microg mL(-1) for APAs. Excellent precision was observed with less than 8.61% RSD. Finally, the method was applied for the detection of ethyl methylphosphonic in aqueous extract of soil sample.

Extraction of acidic degradation products of organophosphorus chemical warfare agents. Comparison between silica and mixed-mode strong anion-exchange cartridges.

The analysis of alkyl alkylphosphonic acids (AAPAs) and alkylphosphonic acids (APAs), the hydrolyzed products of nerve agents, constitutes an important aspect for verifying the compliance to the Chemical weapons convention (CWC). This work devotes on the development of solid-phase extraction method using polymeric mixed-mode strong anion-exchange (Oasis MAX) cartridges for extraction of AAPAs and APAs from water. The extracted analytes were analyzed by GC-MS under full scan and selected ion monitoring mode. The extraction efficiencies of MAX and silica-based anion-exchange cartridges were compared, and results revealed that MAX sorbents yielded better recoveries. Extraction parameters, such as loading capacity, extraction solvent, its volume, and washing solvent were optimized. Best recoveries were obtained using 1 mL of acidic methanol (0.1 M), and limits of detection could be achieved up to 5 x 10(-4) microg mL(-1) (in SIM) and 0.05 microg mL(-1) in full scan mode. The method was successfully employed for the detection and identification of alkylphosphonic acids present in soil sample sent by the Organization for Prohibition of Chemical Weapons (OPCW) in the official proficiency tests.