Une cause traumatique est retrouvée dans 20 à 25 % des cas des dé

Une cause traumatique est retrouvée dans 20 à 25 % des cas des décès liés au sport. Concernant les causes non traumatiques, les pathologies

cardiovasculaires sont les plus learn more fréquentes (75 à 80 %) [1]. Les autres causes non traumatiques sont dominées par le « coup de chaleur ». Plus rarement sont rapportées des causes neurologiques (épilepsie, rupture d’anévrysme) et pulmonaires (embolie ou état de mal asthmatique). Des complications de certaines hémoglobinopathies comme la drépanocytose et la prise de médicaments sont aussi parfois retrouvées. Le commotio cordis est très rare (≤ 3 %). Il est lié à un traumatisme thoracique (coup dans les sports de combat, balle dans le baseball ou puck de hockey) dans la région para-sternale basse gauche. Il concerne essentiellement les jeunes sportifs au thorax très dépressible. L’impact

peut induire un bloc atrioventriculaire complet ou une tachycardie ventriculaire dégénérant en fibrillation [2]. Sa prévention repose sur le port de matériel de protection adapté [2]. À partir des études actuellement disponibles, il n’est pas possible de proposer des statistiques précises sur la prévalence ou l’incidence des morts subites cardiovasculaires liées au sport [3], [4], [5], [6], [7], [8], [9] and [10]. En effet, les données à notre disposition proviennent pour DAPT in vitro la plupart des États-Unis (état du Minnesota surtout) et d’Italie (région de Vénétie), d’études très hétérogènes du point de vue de la méthodologie, concernant plus les compétiteurs que la population sportive générale, avec des modes de recueil (régional ou national, registres ou consultations des médias) variés, rarement associées à une autopsie systématique et jamais avec analyse génétique [11], [12], [13] and [14]. Il est bien démontré que les hommes sont largement plus concernés que les femmes (sex-ratio de 3 à 20 ! et en moyenne 7 à 9), que la pratique de la compétition est plus à risque (risque relatif 2,5 à 5 par rapport au sujet non entraîné apparié) que l’activité sportive modérée et que

les sportifs Afro-Caribéens sont plus touchés que les Caucasiens. Concernant les incidences, des chiffres peuvent être proposés. Ils sont sûrement Thiamine-diphosphate kinase sous-estimés. Chez les jeunes compétiteurs (12–35 ans), elle est comprise entre 1/25 000 à 1/50 000 (0,4–0,7/100 000 chez le sédentaire) dont 33 % de moins de 16 ans. Après 35 ans, elle est plus fréquente et varie entre 1/15 000 et 1/25 000. En France, une étude régionale prospective et un registre national ont estimé le nombre de morts subites liées au sport dans la population générale à au moins 1000 par an, soit près de 3 par jour [6] and [9]. L’augmentation de l’incidence de ces accidents, récemment rapportée, peut s’expliquer par un recueil plus exhaustif et l’augmentation exponentielle du nombre de pratiquants et de compétiteurs chez les vétérans (400 coureurs au marathon de New York en 1976 vs 48 000 en 2009 !). Au total, répétons que la mort subite lors de la pratique sportive reste très rare.

p i , but none of the animals showed rise in body temperature (da

p.i., but none of the animals showed rise in body temperature (data not shown). At both 3 and 14 d.p.i. there was no virus replication in the brains, spleen and intestine (data not shown). This study confirmed the attenuated phenotype of a A/17/California/2009/38 pandemic LAIV candidate in a ferret model. The results of immunogenicity study showed that a single dose of pandemic LAIV was sufficient to induce adequate immune responses against the wild type strain. Moreover, vaccinated animals

proved to be protected against challenge with a virulent wild type pandemic H1N1 virus (Table 2). The monovalent LAIV contained 7.0 log EID50 Baf-A1 nmr per 0.5 ml dose for adults and 6.5 log EID50 for children. Following successful preclinical studies, a Phase I/II randomized, controlled, double-blind clinical study was carried out in 120 adults aged 18–60 years randomly divided into groups to receive

either the vaccine (100) or the placebo (20) administered intranasally in two doses given at 21 days apart. Standard haemagglutinin inhibition (HAI) assays were performed and influenza virus-specific serum IgG and IgA antibodies in nasal swabs were tested by enzyme-linked immunosorbent assays (ELISA) using whole purified virus at 16 HAU per 0.05 ml for absorption. No clinically significant solicited adverse events attributable to the LAIV RAD001 were detected seven days after vaccination (Table 3). The few reactions reported were of short duration and without sequelae. HAI and ELISA tests were also used to determine the serological

response in 66 adult subjects (Table 4). Although post-vaccination serum HA antibody titres were low, cumulative data from both assays resulted in 42.5% and 70.2% conversion after the first and second inoculation, respectively. Peripheral blood mononuclear cells were obtained for analysis by cytokine tests at various times following the first and second vaccination from a limited number of volunteers (16 vaccinees and 9 placebo recipients, respectively). Fig. 1 represents post-vaccination Cell press changes (n-fold) of cellular immune response mediated with virus-specific CD3+CD4+IFNγ+ and CD3+CD8+IFNγ+ memory T cells in volunteers who received LAIV and placebo. After revaccination, the mean increases of both CD4+ and CD8+ memory cells were significantly higher in vaccinated volunteers compared with the placebo group. Interestingly, the same effect of vaccination was also observed in vaccinees without reliable conversions of HAI antibody titres. Even after a single vaccination, the rate of volunteers with a significant increase of these cells in the blood (i.e. results exceeding 2 standard deviations of placebo mean value) was 37.5% (CD8+) and 75.0% (CD4+). After revaccination, the percentage of individuals with significant rises in CD8+ and in CD4+ cells was 68.8%. HAI test results in children were much higher, i.e. 41.4% and 83.

To construct the miR-558 expression plasmid, a precursor of the m

To construct the miR-558 expression plasmid, a precursor of the miR-558 sequence amplified from HepG2 genomic DNA was cloned into the pRNA-U6.1/Neo-siFluc vector. The 3′-UTR region of CXCR5 including the rs3922 locus was

amplified and inserted downstream of the luciferase reporter gene in pGL3-Control Vector. The luciferase reporter plasmid carrying an “A” allele in rs3922 was marked as pGL3-3922A-luc, while the pGL3-3922G-luc contains the SNP “G”. HEK 293T cells were seeded into 12-well plates. Twenty-four hours later, the cells were co-transfected with 1.5 μg of miR-558 expression plasmid or U6 control vector and 50 ng pGL3-3922 luciferase http://www.selleckchem.com/products/Pazopanib-Hydrochloride.html vectors. The pRL-TK (25 ng) plasmid was also transfected as a transfection efficiency control. The luciferase activity in each well was quantified 24 h after transfection using a dual luciferase reporter kit (Promega, Madison, WI, USA) according to the manufacturer’s instructions. In an additional experiment only luciferase vector (pGL3-3922A-luc or pGL3-3922G-luc) and pRL-TK plasmid were co-transfected, without the miR-558 expression plasmid or U6 control vectors. The majority of most experimental conditions were the same in this case except the quantity of pGL3-3922 vector added was Akt inhibitor 100 ng per well. For each SNP, the association between response statuses to HBV vaccine and various genotypes or allelotypes was estimated

by the chi-square test using SAS version 9.1.3 (SAS Institute, Inc., Cary, NC, USA). The Hardy–Weinberg equilibrium (H–W equilibrium) was calculated based on the control group using Haploview version 4.2 software [23]. Linkage disequilibrium

(LD) analysis and haplotype construction were carried out with the same software. Specific parameters were set as previously published almost [4]. P-values, odds ratios (OR), and 95% confidence intervals (95% CIs) were obtained for correlation analysis. A P-value < 0.05 was taken to be statistically significant. A total of 24 SNPs from TfH associated molecules were analyzed in the 20 non-responders and 45 responders. The genotype and allele frequencies of all the SNPs in the study and control groups are listed in Supplementary Table 3. The H–W equilibrium was evaluated in the normal response group and two SNPs (rs3092945, rs715762) in CD40L were excluded from the analysis due to disequilibrium (P < 0.001). Of the remaining 22 SNPs, four (rs3922, rs676925, rs497916 and rs355687) showed significant associations with the immune response triggered by HBV vaccination (P < 0.05, Table 1). Three of these were located in the CXCR5 gene: rs3922 (in 3′-UTR), rs676925 (in 3′-UTR) and rs497916 (in intron), while the fourth one rs355687 was located in the intron of CXCL13. As collected by the international HapMap project, the distributions of these 4 SNPs in different populations were summarized in the Supplementary Table 4.

1) [28] This study was conducted in 4 Latin American countries (

1) [28]. This study was conducted in 4 Latin American countries (Mexico, Costa Rica, Guatemala, and Brazil) where OPV was given at ∼2, 4, and 6 months of age. RotaTeq® was given either

click here at the same time as OPV or 2–4 weeks before OPV. After vaccination with the full 3-dose RotaTeq® series, antirotavirus IgA GMC was 47% lower when RotaTeq® was given with OPV (155 U/mL; 95% CI = 126–190) compared to when RotaTeq® was given separately from OPV (293 U/mL; 95% CI = 249–345), with non-overlapping confidence limits. Seroconversion (defined as ≥3-fold rise in serum antirotavirus IgA level) was 5% lower without OPV (93%) than with OPV (98%). Country specific data were not provided. The experience with current and previous rotavirus vaccines provides several important insights relevant for understanding the potential impact of OPV on rotavirus vaccination. The immune response to the first dose of rotavirus vaccination given concomitantly with the first dose of OPV has almost

always been lower than when vaccine was given without OPV, indicating interference with immune response selleckchem to rotavirus vaccination by OPV. However, a review of the older rotavirus vaccines (i.e., not in current use) suggest that OPV’s negative effect on the first dose of rotavirus vaccination has generally been overcome by administration of subsequent rotavirus vaccine doses [20], so that comparable immune responses were seen after the full vaccine series among infants receiving vaccine with or without OPV. The three-doses of RotaTeq® are to be given with the routine EPI schedule, which is at 2, 4, and 6 months in the Americas, but at somewhat younger ages of 6, 10, and 14 weeks in Africa and Asia. For the two-doses of Rotarix™, the WHO recommended that the vaccine should be given with the first two doses of the EPI schedule at 6 and 10 weeks of age [32]. The interference from OPV is likely to have a greater negative impact on efficacy of and rotavirus vaccine

during the first EPI visit at 6 weeks of age, when circulating maternal antibodies are also high and are known to also interfere with vaccine take [13], compared to the second and third EPI visits at 10 and 14 weeks of age. Indeed, an earlier immunogenicity study in South Africa demonstrated better immune responses after two doses of the monovalent rotavirus vaccine, RIX4144, at 10 and 14 weeks of age compared to 6 and 10 weeks of age [26]. Therefore, more evaluations are needed in Asia and Africa to assess the efficacy of Rotarix™ when administered at the WHO-recommended 6–10 week schedule compared to alternative schedules such as 10–14 or perhaps 3 doses at 6–10–14 weeks of age. The key question is whether the impact of OPV on the immune response to rotavirus vaccines translates to a reduced protective efficacy, as measured immune responses to rotavirus vaccination do not necessarily correlate with efficacy.

06 (95% CI: 1 05–1 08) Age over 35 years, residing in urban area

06 (95% CI: 1.05–1.08). Age over 35 years, residing in urban areas or in the Auckland region, riding in a bunch, using a road bike and history of a crash at baseline predicted a higher risk whereas being overweight or obese, cycling off-road and using lights in the dark lowered the risk. Bicycle commuting, however, did not increase the risk. There were 10 collisions per 1000 person-years or 38 collisions per million hours spent road cycling per year (Table 4). The adjusted HR for one click here hour increase in average time spent

cycling each week was 1.08 (95% CI: 1.05–1.12). Due to a very small number of events, “overweight” and “obese” categories were combined and helmet use was excluded in the multivariate models. Residing in urban areas, riding a road bike and having a crash history were associated with an increased risk. There were 50 crashes per 1000 person-years (Table 5). The risk was lower in university graduates, overweight or obese

cyclists and less experienced cyclists but higher in those who cycled in the dark or in a bunch and those who had a crash history. The effect estimates mentioned above were similar to those obtained from complete case analyses. Potential misclassification of crash outcomes during the linkage process may underestimate the actual incidence rate and may bias the hazard ratios to the null (Appendix A). Likewise, potential misclassification of exposures check details (due to changes over time) may underestimate the risk estimates in most cases (Appendix B). In this study, cyclists experienced 116 crashes attended medically or by police per 1000 person-years, of which 66 occurred on the road and 10 involved a collision PD184352 (CI-1040) with a motor vehicle. There were 240 on-road crashes and 38 collisions per million hours spent road cycling and the risk increased by 6% and 8% respectively for one hour increase in cycling each week.

After adjusting for all covariates, participants’ age, body mass index, urbanity, region of residence, cycling off road, in the dark or in a bunch, type of bicycle used and prior crash history predicted the crash risk with variations in effect estimates by crash type. This is one of the very few prospective cohort studies involving cyclists and used record linkage to obtain objective information on bicycle crashes from multiple databases. This resource efficient method of data collection was also designed to minimise potential biases associated with loss to follow-up (Greenland, 1977) and self-reports (af Wåhlberg et al., 2010, Jenkins et al., 2002 and Tivesten et al., 2012). While emigration during follow-up is a potential issue in using the linked data, this accounted for less than 2% of the participants resurveyed in 2009 and may not substantially influence outcome occurrences (Kristensen and Bjerkedal, 2010).

aeruginosa (0 0156 mg/ml), K pneumonia (0 0156 mg/ml),

B

aeruginosa (0.0156 mg/ml), K. pneumonia (0.0156 mg/ml),

B. subtilis (0.0156 mg/ml) and activity against E. coli (0.0156 mg/ml) is comparable with the standard antibacterial agent Tetracycline. Compound 2g shows good activity among all the compounds against S. typhi (0.0625 mg/ml). In antifungal assay, all the compounds 2a–j displayed good antifungal activities against fungi A. flavus (0.0625–0.46 mg/ml), A. fumigatus (0.125–3.75 mg/ml). Compounds 4a–i demonstrated moderate results against the fungi A. niger (0.125–7.5 mg/ml). The evaluation of the antioxidant effects of the newly synthesized compounds having different concentrations were examined by well documented in vitro assay i.e. DPPH free radical scavenging assay. Antioxidant reacts with a stable free radical DPPH and converts it to 2,2-diphenyl-1-picryl hydrazine. The degree of Galunisertib solubility dmso decoloration indicates the scavenging potentials of the compound. The percentage (%) DPPH activities

of all the synthesized compounds have been shown in Table 2. The investigation of DPPH assay reveals that the, BMS 354825 compound 2g shows good activity comparable with the standard compound Ascorbic acid. Remaining compounds exhibited moderate to good activities as shown in Table 2. A series of novel formazans containing 3,4-dimethylpyrrole moiety were synthesized and their structures were confirmed by IR, 1H & 13C NMR, mass spectroscopy. The antimicrobial and antioxidant activities of the new compounds were evaluated. The results of preliminary bioassays of derivatisation of Schiff bases to formazan indicate that a number of these molecules exhibits moderate to good antibacterial, antifungal and antioxidant activities some of which are comparable to standard used in this study. The outcome indicates that there is a good scope for evaluation of this class of compounds as potential leads towards antimicrobial and antioxidant agents. All authors have none to declare. JDB is thankful to UGC-SAP for RFSMS Fellowship. “
“Natural products principally medicinal plants have long been prescribed in traditional medicine for centuries for treating different diseases. The significance

of herbs in the management of human ailments cannot be overemphasizing. The repetitive below investigation into the secondary plant metabolites for anti-infective agents has gained consequence because of the alarming increase in the rate of antibiotic resistance of pathogenic microorganism to existing antibiotics. Therefore the need to develop proficient, safe and inexpensive drugs from plant sources is of great importance.1 Antibiotic resistance has become a global concern. There has been an increasing incidence of multiple resistances in human pathogenic microorganisms in recent years, mostly due to haphazard use of commercial antimicrobial drugs commonly employed in the treatment of infectious diseases. This has forced scientists to search for new antimicrobial substances from various sources like the medicinal plants.

N-acylated benzyl carbamate with 86% yield was achieved in 20 min

N-acylated benzyl carbamate with 86% yield was achieved in 20 min of time. Then we examined the reaction conditions in presence of anhydrous cerium Chloride with the same substrates, observed that the reaction learn more was completed within 6 min of time with 95% yield ( Scheme. 2, Entry-1 in Table 2) and decided to go with anhydrous cerium chloride to explore the substrate scope in this case as well. These reaction conditions were success

full while exploring the possibilities with structurally diversed acid anhydrides like propionic, pivalic and benzoic anhydrides. We have examined the same reaction conditions to find out the applicability in case of secondary carbamates like amino acid carbamates and amine carbamates and found positive results. All the results regarding the N-acylation of carbamates were mentioned in Table 2. Synthesized compounds were screened check details for their antifungal activity by anti Malassezia in vitro liquid broth culture in high-throughput assay format for anti-dandruff activity testing against two virulent organisms M. furfur and M. pachydermatis MF-ATCC44338 MP-ATCC42757 were the corresponding strains. The compounds were tested in four replicates in the concentration range of 200 uM, 180 uM, 160 uM, 140 uM, 120 uM, 100 uM, 75 uM, 50 uM,

25 uM, 10 uM and 1 uM by incubating them for stipulated time period of 72 h and taking their growth observations in the form of optical density at 600 nm wavelength at different time intervals. The growth in the treated wells was compared with the growth in the untreated wells. Ketoconazole was used as control, among heptaminol the compounds

screened 2a, 2i and 4a showed activity than the standard antifungal drug i.e. Ketoconazole, corresponding results were mentioned in Table 3. We have developed a novel and efficient method for of N-acylation of sulfonamides and carbamates with carboxylic acid anhydrides under solvent free and mild reaction conditions in presence of cerium (III) chloride. Synthesized compounds were evaluated for their antifungal activity against M. furfur and M. pachydermatis. Three compounds 2a, 2i, and 4a showed very good activity against both the organisms, for the first time N-acyl sulfonamides and carbamates class was evaluated as potential anti-Malassezia agents. This outcome indicates that there is a good scope for evaluation of this class of compounds as potential leads towards anti Malassezia activity. All authors have none to declare. “
“Tissue engineering is very fast growing scientific area in this era and used to create, repair, and/or replace cells, tissues and organs by using cell and/or combinations of cells with biomaterials and/or biologically active molecules and helps to produce materials which very much resembles to body’s native tissue/tissues. Tissue engineering is the connecting discipline between engineering materials science, medicine and biology.

, 2006) Similarly, a primate study showed that fluoxetine treatm

, 2006). Similarly, a primate study showed that fluoxetine treatment prevented the onset of depression-like this website behaviours and increased the number of newly-born neurons that were at the threshold of maturation within a specific region of the dentate gyrus (anterior region), thus leading to the suggestion that adult hippocampal neurogenesis may contribute to the recovery promoted by

fluoxetine (Perera et al., 2011). On the other hand the antidepressant-like effects of non-monoaminergic based antidepressant-like drugs, such as CRH1 or V1b antagonists, are not affected by inhibition of adult hippocampal neurogenesis (Surget et al., 2011 and Bessa et al., 2009) which is in contrast to many findings with antidepressants that target the monoaminergic system such as fluoxetine and imipramine (Surget et al., 2011, Perera et al., 2011 and Santarelli et al., 2003). Thus, it has been suggested that antidepressant drugs increase adult hippocampal neurogenesis,

independently of their behavioural effects and that antidepressant-induced increases in adult hippocampal neurogenesis might not be the final process in the recovery from stress-induced depressive-like behaviour BMS-354825 purchase (Bessa et al., 2009). The hippocampus can be divided along its septotemporal axis into dorsal and ventral regions in rodents and into anterior and posterior regions in primates, based on their distinct afferent and efferent connections (Fanselow and Dong, 2010). Lesion, optogenetic and electrophysiological studies in rodents suggest that this anatomical segregation results in a dichotomy in the Adenylyl cyclase function of the dorsal hippocampus (dHi) and the ventral hippocampus (vHi) (Fanselow and Dong, 2010 and Bannerman et al., 2004). While the dHi (analogous to the posterior hippocampus in primates) seems to play a preferential role in spatial learning and memory processes, the vHi (analogous to the anterior hippocampus in primates) preferentially regulates anxiety and the response to stress (Fanselow and Dong, 2010, Bannerman et al., 2004 and Moser and Moser, 1998). Since adult hippocampal

neurogenesis has been implicated in processes preferentially regulated by the dHi (spatial learning and memory) and the vHi (stress response), it is possible that adult neurogenesis might be regulated preferentially in the dHi or the vHi, depending upon the stimulus (Tanti and Belzung, 2013 and O’Leary and Cryan, 2014). Indeed, several studies have reported that stress affects several stages of adult neurogenesis, preferentially in the vHi rather than the dHi (Tanti and Belzung, 2013 and O’Leary and Cryan, 2014). Some (but not all) studies also report that antidepressant-induced increases in cytogenesis and neurogenesis occur preferentially in the vHi but not dHi (Tanti et al., 2012, Jayatissa et al., 2006, O’Leary et al., 2012, O’Leary and Cryan, 2014 and Banasr et al., 2006).

In special circumstances like the DPT-hepatitis B-Hib vaccine iss

In special circumstances like the DPT-hepatitis B-Hib vaccine issue, the ACCD requests

external technical assistance to inform recommendations. WHO, for instance, was invited to carry out an independent assessment of causality in the DPT-hepatitis B-Hib and rubella vaccine incidents. The WHO assessment provided an unbiased, second opinion for the Committee to consider. The Committee discussed the findings from both the Expert Committee on AEFI and the WHO assessments – both of which found no conclusive evidence that the DPT-hepatitis B-Hib vaccine caused the deaths – before recommending that the NPI reintroduce the vaccine. Though the decision was not unanimous, the discussions that took place between the Expert Committee on AEFI and WHO further strengthened the capacity of the ACCD to arrive at practical, evidence-based conclusions regarding the future course of action for this vaccine. A similar process was used to respond to the rubella incident, check details which helped the ACCD to counter the widely held belief among the public

and health worker trade unions that it was not anaphylaxis but the inferior quality of the vaccine that caused the death of the child. The ACCD can also recommend health system improvements that will help ensure the success of immunization and other disease control measures. As demonstrated during the DPT-hepatitis B-Hib incident, one selleck screening library drawback in investigating deaths among vaccine recipients in Sri Lanka was the absence of a definitive cause of death, even for deaths in which post mortems had been performed. This was attributed to the fact that Judicial Medical Officers (JMOs), forensic experts who perform autopsies and determine cause of death in homicide cases, conducted these post mortems, but had not been trained to look for pathological causes. The ACCD was able to rectify this by mandating that consultant JMOs use a standardized autopsy protocol when conducting post mortem examinations of all deaths suspected to be immunization-related. A summary of the data required and questions to be answered before the ACCD makes a recommendation about a new vaccine is shown in Fig. 2. To second formulate policy recommendations regarding the

introduction of new vaccines, the ACCD requests a set of data from the Epidemiology Unit. The Unit then appoints a working group, consisting of experts from Ministry of Health agencies, major hospitals, universities and the private sector, to help gather and analyze relevant data concerning the disease and vaccine in question. The Epidemiology Unit may also request technical or financial support from international partners for the collection or analysis of data, in the form of, for instance, an expert, such as a health economist, financing to conduct a local clinical trial, or laboratory training for surveillance studies. The compilation of data on the burden of the disease in question in Sri Lanka is a necessity before the ACCD can approve the introduction of any new vaccine.

HPLC data acquisition and processing

HPLC data acquisition and processing IOX1 was performed by Shimadzu LC Solutions Ver 1.23 SP 1 software. PZA belongs to the basic class of drugs due to its amide functional group. Therefore adjusting the pH of mobile phase to the acidic side ionizes the PZA present in plasma thereby leading to poor recovery. In order to extract the un-ionized form of the drug, it is imperative to adjust the pH to the alkaline side, however, alkaline mobile phase characteristics causes deterioration of the bonded phase in the column due to alkaline hydrolysis of end-capped silica. Compared to acid catalyzed hydrolysis, the hydrolysis of end-capped

silica in alkaline conditions is usually very rapid. Therefore experiments were performed using potassium dihydrogen phosphate in a limited range of pH 7.0–8.0. The response was checked at the detector using a connector (without the column). A pH value of 7.4 ± 0.1 gave maximum Selleck ABT-199 response for the analyte at 268 nm. The run time of analysis was higher when a longer reverse phase column (250 × 4.6 mm i.d,) was used. The resolution

between the peaks was decreased and peaks were not of acceptable peak shape when the experiment was performed using a shorter column (50 × 4.6 mm i.d,). However better resolution, less tailing and high theoretical plates were obtained with Phenomenex column C18 150 × 4.6 cm 5 μm column. The mobile consists of 15:85 v/v methanol and 10 mM potassium dihydrogen phosphate (pH 7.4). The flow rate of the method was 1.0 ml/min. The column temperature was maintained at 25 °C. At the reported flow rate peak shape was excellent,

however, increasing or decreasing the Astemizole flow rate increased the tailing factor and resulted in poor peak shape and in decreased resolution between the drug and internal standard. There was no interference in the drug and the internal standard, from the extracted blank. The peak shape and symmetry were found to be good when the mobile phase composition of 15:85 v/v was used with better resolution of the drug and internal standard. Increasing the organic portion of the mobile phase caused PZA to elute with high tailing and also merging of the peaks for PZA and MTZ. A mobile phase containing aqueous portion greater than 85% led to very late elution and very poor peak shape for MTZ. The peaks were also broad and had unacceptable asymmetry factor. Extraction methods were initially attempted using protein precipitation technique. Organic solvents such as acetonitrile and/or methanol were used as reagents for protein precipitation.13 Initial experiments of protein precipitation were done using 1:3 ratio of plasma:organic solvents. The recovery of the PZA was poor while that of the internal standard was relatively unchanged as compared with liquid–liquid extraction. Since the noise effects in solid phase extraction (SPE) method are similar to that of liquid–liquid extraction, the final analysis was carried out using liquid–liquid extraction (LLE).