The number of probes per cell was calculated based on the total photon count with the subtraction of the background count. The calibration of the set-up was performed by collection of luminescence light from a thin layer of the probes solution excited directly by the laser beam at the right angle from the bottom of a thin fused silica substrate. The microscope field of view in these experiments was 14 × 14 μm2. To achieve homogeneity of the excitation beam, the beam was http://www.selleckchem.com/products/nutlin-3a.html passed through a 0.32 cm2 diaphragm. The pulse energy was measured after the diaphragm (0.32 mJ pulse−1).

This allowed a reliable determination of the laser light fluence. Measured volume of the probes solutions (1.12 mM Probe 1-Eu3+ or 0.107 mM Probe 4-Tb3+) in glycerol was placed on the top of the substrate and spread upon the surface with a cover slip (the spot area of 3.80 cm2 and the thickness of the layer of 2.63 μm). The luminescence Etoposide light intensity was calculated based on the photon fluence, the absorption cross-sections of the probes at 351 nm (2.1 × 10−17 cm2 molecule−1 and 3.6 × 10−17 cm2 molecule−1 for probes Eu3+ and Tb3+respectively), the luminescence quantum yield (0.167 for Eu3+[14], and ca. 0.45 for Tb3+ probe), and the total number of probes in the field-of-view area. This was compared with the total

number of photons counted in the image. This procedure allowed determination of the calibration coefficients, which lump sum the solid angle of light collection of the objective lens, the microscope throughput coefficient, the photocathode quantum efficiency, as well as the photon counting efficiency. The average number of the probes per externally labeled E. coli cells determined in this way was 2.1 × 105 and 2.9 × 105 for Eu3+ and Tb3+ probes,

respectively. Externally labeled CHO cells were prepared in a similar manner. The cells were labeled with ADP ribosylation factor avidin conjugates carrying multiple Eu3+ chelates of probe 1 with an average 1.6 × 107 probes per cell. The detection of light emission of a lanthanide chelates and their conjugates with avidin as well as of BODIPY-modified avidin was performed in a measuring cell 150 μl) in a buffer containing 10 mM Hepes pH 8.0. Water-based or deuterium oxide-based solutions were used. In our previous study [15], we found a convenient modification reaction for the cs124CF3 fluorophore, which allows introduction of the crosslinking groups at N1 position. Here we performed the same reaction with parent cs124 compound in order to obtain probe 4 (Fig. 1). Similarly to corresponding trifluoro-derivative, alkylation of cs124 fluorophore by bifunctional biphenyl compound produced alkylation product at N1 with high yield (Fig. 2). Notably, alkylation proceeded almost exclusively at N-1 of the quinolone ring, while the same reactions with ethyl ester of 4-toluenesulfonic acid or with 1-iodo-3-azidopropane yielded detectable amount of O-alkylated products (15).

Yellow gummy solid, 1H NMR (400 MHz, CDCl3): δ 2.33 (s, 3H), 2.68 (brs, 4H), 3.04 (brs, 4H), 3.66 (s, 2H), 3.77 (s, 3H) 3.88 (s, 3H), 6.91–6.93 (m, 1H), 7.09–7.14 (m, 2H), 8.21 (s, 1H). 13C NMR (100 MHz, CDCl3): δ (ppm) 164.02, 156.19, 151.42, 148.6, 133.94, 127.43, 127.35, 126.09, 125.00, LY2109761 124.34, 118.54, 62.68, 59.83, 53.32, 51.30, 13.24, 11.00; MS (e/z): 379,

381 (M−, M+). Anal. calcd. for C19H23Cl2N3O: C, 60.00; H, 6.10; Cl, 18.64; N, 11.05; O, 4.21. Found: C, 60.11; H, 6.05; N, 11.15. Pale yellow gummy solid, Mass (e/z): 1H NMR (400 MHz, CDCl3): δ 2.06–2.27 (m, 2H), 2.27 (s, 3H), 2.69 (brs, 4H), 3.05 (brs, 4H), 3.36 (s, 3H), 3.58 (m, 2H), Selleck Wortmannin 4.10 (t, 3H), 6.69 (d J = 5.6 Hz, 2H), 6.91–6.93 (m, 1H), 7.09–7.14 (m, 2H), 8.30 (s, 1H). 13C NMR (100 MHz, CDCl3): δ (ppm) 163.01, 157.09, 151.82, 149.6, 134.25, 128.42, 127.43, 126.28, 125.12, 124.45, 118.65, 77.53, 71.42, 64.51, 62.82, 59.94, 53.45, 51.41, 11.28; MS (e/z): 423, 425 (M−, M+). Anal. calcd. for C21H27Cl2N3O2: C, 59.44; H, 6.41; Cl, 16.71; N, 9.90; O, 7.54. Found: C, 59.56; H, 6.34; N, 9.98. Light brown colour syrup. 1H NMR (400 MHz, CDCl3): δ 2.32 (s, 3H), 2.69 (brs, 4H), 3.05 (brs, 4H), 3.73 (s, 2H), 4.36–4.4.42 (q = 3H), 6.64 (d, J = 8 Hz, 1H), 6.91–6.93 (m, 1H), 7.01–7.05 (m, 4H), 8.36 (d, J = 5.6 Hz, 1H). 13C NMR (100 MHz, CDCl3): δ (ppm) 163.13, 157.18,

151.91, 149.62, 134.31, 128.52, 127.32, 126.34, 125.21, 124.52, 122.12, 118.72, 85.72, 62.99, 60.08, 53.65, 51.61, 11.45; Mass (e/z): 433, 435 (M−, M+). Anal. calcd. for C19H20Cl2F3N3O: C, 52.55; H, 4.64; Cl, 16.33; F, 13.12; N, 9.68; O, 3.68. Found: C, 52.66; H, 4.57; N, 9.78. Pale yellow colour syrup. 1H NMR (400 MHz, CDCl3): δ 2.76 (brs, 4H), 3.07 (brs, 4H), 3.77 (s, 2H), 3.77 (s, 3H) 3.88 (s, 3H), 6.81 (d, J = 7.6 Hz, Rebamipide 1H), 6.92–6.94 (m, 1H), 7.01–7.14 (m, 2H), 8.27 (d, J = 5.6 Hz, 1H). ); 13C NMR (100 MHz, CDCl3): δ (ppm) 158.6, 151.3, 145.6, 127.4, 124.3, 118.5, 106.7, 77.5, 77.17, 76.8, 61.08, 58.12, 55.61, 53.27, 51.13; MS (e/z): 381, 383 (M−, M+). Anal. calcd. for C18H21Cl2N3O2: C, 56.55; H, 5.54; Cl, 18.55; N, 10.99; O, 8.37. Found: C, 56.69; H, 5.47; N, 11.08. Pale yellow colour syrup.

An approximation of lifetime cost was obtained by multiplying the average annual cost by the estimated average survival time for patients with incident CC in each country over the 5 years post diagnosis. It was assumed that a cancer patient

alive for 5 years post diagnosis is cured and hence without any treatment and costs associated. The average survival time was estimated for each country using data on the number of annual incident cases and estimates Volasertib order of 5 year prevalence reported by Globocan 2008 [1] as follows: (5⁡ years prevalent cases/incident cases×5)×5=average survival over the 5 years post diagnosis(5⁡ years prevalent cases/incident cases×5)×5=average survival over the 5 years post diagnosis Costs for CC treatment were expressed in local currency and updated to 2011 values using the country-specific Consumer Price Index reported by the World Bank for each country [20]. Estimated survival times and lifetime costs are shown in Table 1. The potential annual effect of HPV vaccination on the burden related to CIN 2/3 at vaccination steady state was estimated in two countries: Italy and Malaysia, randomly selected based on data availability. The method used is identical to the one used to estimate the vaccine impact

on CC cases and deaths. The number of CIN2/3 cases prevented with vaccination irrespective of HPV type, the expected number of HPV-16/18 related CIN2/3 avoided by vaccination cases as well as the difference between the two were estimated. Angiogenesis inhibitor Vaccination coverage was assumed to be 80% in both countries. The prevalent annual numbers of CIN2/3 lesions prior to the introduction of vaccination for Italy and Malaysia were retrieved from literature (Table 2) [5] and [21]. The vaccine effectiveness

those against CIN2/3 lesions irrespective of HPV type was assumed at 64.9% based on the VE reported against CIN2+ lesions, irrespective of HPV type in the HPV-naïve1 TVC from the end-of-study results from the PATRICIA trial [9]. Vaccine effectiveness against CIN2/3 lesions related to HPV-16/18 was estimated based on the effectiveness against HPV-16/18 CIN2/3 lesion and the proportion of CIN2/3 related to HPV-16/18. The vaccine effectiveness against HPV-16/18-related CIN2/3 lesions was assumed to be 100%, based on VE against CIN2+ causally related to HPV-16/18 reported from the end-of-study from the PATRICIA trial among the HPV-naïve1 TVC [9]. The proportion of CIN2/3 related to HPV type-16/18 was calculated based on the HPV-16/18 distribution reported for high-grade cervical lesions in the ICO HPV Information Centre database for each country [2] (Table 2). The expected CIN2/3-related treatment costs potentially offset by HPV vaccination was estimated assuming that 100% of CIN2/3 lesions prevented by vaccination would be treated. The offset on treatment costs was estimated by multiplying the number of cases potentially prevented by the CIN2/3 treatment unit cost.

The Committee also established a sub-committee for the investigation of vaccine-related injuries, which was separated from the KACIP

and became the Advisory Committee on Vaccine Injury Compensation in 2003. Committee members are appointed to 2-year terms that all begin at the same time, and thus a new committee is formed every 2 years. However certain officials, who serve as a result of their position within the government will remain on the Committee for as long as they remain in their position (see next section). Despite this intention, the duration of the current – seventh – committee, which was formed in October 2007, has been extended selleckchem to a third year, because of the many issues it has been dealing with that still need to be resolved. This is the first time that the Committee’s term has been extended and the terms will go back to 2 years

in 2010. Among the items on the agenda of the current committee have been: a review of national immunization strategies; the control of measles; how to control a hepatitis A outbreak; the control of varicella and mumps; whether to change the strain of Bacillus Calmette–Guérin (BCG) vaccine and route of administration (from intradermal to transdermal); and the issue of subsidizing the cost of Expanded Program of Immunization (EPI) vaccines provided through the private sector, through which the majority of immunizations in Korea are given. Based on a recommendation by the KACIP, the Government has decided to partially SP600125 subsidize the

cost of all EPI vaccines administered at private health facilities that agree to participate in this program, starting in 2009 (with parents now paying 70% instead of 100% of the vaccine cost). The KACIP consists also of a Chairperson and specialists in internal medicine, paediatrics, obstetrics, microbiology, preventive medicine and nursing. The Committee also includes a representative from a consumer group, the Director of Disease Prevention at the Korea Centers for Disease Control and Prevention (KCDC), and the Director of Biologics at the Korea Food and Drug Administration (KFDA). Apart from the two government officials mentioned above, all other members usually come from the affiliated organizations shown in Fig. 1, which each nominate one member. The total number of Committee members is usually around 15. The Secretariat of the Committee is within the KCDC, which funds, organizes and prepares for the meetings, and at whose headquarters the meetings are held. The Chairperson rotates every term (i.e., 2 years) and can be selected from any field or affiliated organization. Over the years, Committee members have made recommendations to include more female members, representatives from civil society, and people from rural areas, though to date there are no minimum requirements or quotas for representation of these groups.

and GlaxoSmithKline. Several other indigenously manufactured rotavirus vaccines are in development in India, some of which are in late stages of clinical testing. With an effective, indigenously produced rotavirus vaccine on the near-term horizon, India, which singularly accounts for almost one fifth of the world’s burden of rotavirus deaths in children [2], is poised to have a new tool in the arsenal of interventions to reduced morbidity and mortality from childhood diarrhea. To help assess

the public health value of the vaccine, understanding the current rotavirus disease burden and epidemiology, circulating strains, and economic burden of rotavirus in India is important. This supplement contains papers summarizing the most up-to-date data on these issues. In addition, the supplement addresses areas relevant for post-introduction monitoring of rotavirus vaccine, including potential safety concerns associated with see more other rotavirus vaccines such as intussusception, a condition in which one portion of the bowel telescopes into another causing a blockage. Finally, this supplement contains papers looking at the performance of rotavirus vaccines, both the indigenous and internationally available vaccines, in India and explores strategies to improve vaccine

performance. This PF-02341066 manufacturer collection of papers will help provide a complete picture of rotavirus disease in India and the potential for a rotavirus vaccination program, and also set the platform to assess the impact of vaccines post-introduction. Rotavirus persists as a major cause of severe acute diarrhea in Indian children. By 5 years of age, an estimated 1 out of every 344 Indian children will die

from rotavirus diarrhea, 1 in every 23–46 children will be hospitalized for rotavirus diarrhea, and 1 in every 6 to 12 children will have an outpatient visit due to rotavirus diarrhea [3]. This translates into 78,500 deaths, 872,000 hospitalizations, over 3.2 million outpatient visits and 11.37 million diarrhea episodes due to rotavirus in children <5 years of age each year in India [3]. Most previous disease burden estimates have provided figures for mortality and hospitalizations alone, and hence the availability of these updated estimates, which include outpatient visits (-)-p-Bromotetramisole Oxalate and diarrheal episodes managed at home, will provide a tool to better assess the health and economic burden of disease that might be alleviated by rotavirus vaccination. Rotavirus causes a significant proportion of the severe health burden due to diarrhea. Sentinel hospital-based surveillance, often conducted as part of the Indian Rotavirus Surveillance Network, found the proportion of diarrheal hospitalizations among children <5 years of age associated with rotavirus ranging from 26% in Vellore, 35% in Pune, 38–40% in Delhi, 50% Trichy, and 53% in Kolkata [4], [5], [6], [7] and [8] (Fig. 1).

AS and HCQ Sulphate were obtained as gift samples from Indian Printed Circuit Association India. Sodium chloride (NaCl), potassium dihydrogen orthophosphate, alcohol and HCl were analytical grades as required and were obtained from Qualigens, India. The solubility of AS and HCQ was studied in various hydrophilic and lipophilic solvents and pharmaceutical buffers. In each case, 25 mg of AS and HCQ were mixed separately with 25 ml of respective solvents and shaken gently at room temperature for 10 min and the degree of solubility was observed. A definite quantity of drug powder (AS) (10 mg) was kept in glass bottles and these bottles are stored at 2–8 °C/60%

Relative humidity (RH), 25 °C/65% RH, 40 °C/75% RH and 50 °C/60% RH in a humidity LY294002 purchase control oven. Drug analysis was carried out after time interval of 24 h after, 1 week, 3 weeks and 5 weeks by colorimetric method.18 Drug degradation that involves reaction with water is called hydrolysis. Hydrolysis is affected by pH, buffer salts, ionic strength, solvent, and other additives such as complexing agents, surfactants, and excipients.19 and 20 AS drug powder (10 mg) was kept in amber glass vials containing phosphate

buffer of different pH ranging from 5.8 to 8.0 and these vials were stored at 2–8 °C and 25 °C. Drug analysis was carried out after time interval of 0 day, 1st week, 3rd Ribociclib research buy weeks and 5th weeks by colorimetric method. The photo reactivity screening of HCQ was performed. To study photochemical

degradation in solid state HCQ drug powder (10 mg, 3 mm thick) was Phosphoprotein phosphatase kept in glass bottles and these bottles were stored at 25 °C in UV cabinet at 240–600 nm. Drug analysis was carried out after time interval of 24 h and 1st week, 3rd week, 5th week.21 To perform compatibility studies HCQ drug powder (10 mg) was dissolved in different solvent system (10 ml) and these volumetric flasks are stored at 4 °C and 30 °C in humidity control oven. Drug analysis was carried out after time interval of 24 h, 1st week, 3rd week and 5th weeks.22 The solubility analysis performed with AS reveals that the compound is maximum soluble in methanol (99% solubility). The solubility analysis performed in ethanol states that as percentage of alcohol increases the solubility increases. The drug was more soluble in methanol than ethanol. The drug was 29.8% soluble in acidic media i.e. 0.1 N HCl. Addition of alcohol in 0.1 N HCl increased solubility, from 29.8% to 98%. The drug had poor solubility in water and normal saline. The analysis in alkaline medium i.e. phosphate buffer saline of alkaline pH range reveals that as the pH increased from pH 5 to pH 7 the solubility increased, while increase in pH beyond 7 decreased solubility. Hence from results it is concluded that alcohol can be used as co solvent to increase solubility of AS (Table 1). HCQ was also analyzed for solubility in various solvents.

The ratio of apoptotic cells was significantly increased, dependent on PPD concentration (i.e., >20 μM, consistent with the above cell proliferative data), compared with control (Fig. 4A; P < 0.01). HCT-116 and SW-480 cells were treated with different concentrations (15, 20, 25, 30, and 35 μM) of PPD for 48 h and the cell cycle was examined by flow cytometry. As shown in Fig. 4B, PPD-induced G1 cell cycle arrest in a concentration-dependent manner in both cell lines (both P < 0.01). HCT-116 cells were selected to perform mRNAs expression profiling analysis on six samples, learn more including three control vehicle treated cells and different concentrations and time points of PPD-treated cells.

We first performed an unsupervised, two-way (genes against samples), hierarchical cluster analysis (HCA). Remarkably, three PPD-treated cell samples (24p20, 48p20, 48p25) clearly grouped into one cluster, while three normal control cell samples also grouped together and formed a cluster (Fig. 5A). 204 genes significantly changed (over 1.5-fold) after PPD treatment. A sub-analysis based 79 genes significantly altered (over 2-fold) (Fig. 5B). 20 of the most upregulated and downregulated genes were compiled based on the microarray data, shown in Table 1 and Table 2. Among the genes that were Alpelisib in vivo significantly altered when treated

with PPD in HCT-116 cells, six downregulated genes (CLSPN, CCNA2, SPAG5, DNM3, DHCR24, DSCC1) and five upregulated genes (BTG1, DDIT4, PDCD4, KLF4, NRP1) were validated by quantitative real-time RT-PCR. The same RNA samples for microarray were used to generate cDNA templates for reverse transcription reactions. The SYBR green-based real-time RT-PCR analysis was then carried out. Consistent found with the microarray data, the 11 selected genes showed the same expression profile as the microarray data presented (Fig. 5C and D). We performed gene network analysis using the 204 significant genes from our microarray analysis through the Ingenuity Pathway Analysis (IPA). A bar plot presenting ten classic

pathways related to tumorigenesis is shown in Fig. 6A. Among them, apoptosis, proliferation, and angiogenesis were significantly induced. This is consistent with our in vitro data, suggesting that PPD is probably involved in cancer cell growth by modulating these processes. The selected regulatory cell death pathway gene network is shown in Fig. 6B, in which 23 affected genes of this network were either upregulated or downregulated after PPD treatment. Among the genes, DR4 and DR5 are important members of the tumor necrosis factors (TNF) family. It appears that HCT-116 cell apoptosis was induced after PPD exposure by the interaction of p53 and DR4/DR5, and suggests that the TRAIL pathway was associated with the PPD activities. CRC is one of the most common cancers worldwide (18).

Le recours aux techniques neurochirurgicales de section (drezotomie, radicellectomie sélective postérieure, intervention de Nashold, cordotomie antérolatérale) ou de stimulation (stimulation cordonale postérieure, stimulation corticale) est exceptionnel en situation palliative avancée. Les

recommandations formalisées d’experts de la SFAR et de la SFETD, publiées en 2013, portent notamment sur les techniques analgésiques locorégionales dans la douleur chronique cancéreuse, entre autres pathologies [22]. La prise en charge de la douleur nécessite d’avoir de bonnes connaissances théoriques sur les maladies causales, l’évaluation des caractéristiques douloureuses, les propriétés pharmacologiques et les effets indésirables potentiels des médicaments à prescrire pour obtenir un soulagement (antalgiques et co-antalgiques), mais aussi des connaissances pratiques MK0683 molecular weight sur les techniques et soins applicables en parallèle et sur les thérapeutiques non médicamenteuses. À côté de la connaissance et du savoir-faire scientifiques, la relation en soins est une dimension qui prend ici toute sa place pour un savoir-être auprès du patient douloureux. L’écoute

attentive sera l’un des éléments-clés de la prise en charge de la douleur du cancer : écouter la plainte douloureuse du malade nécessite de la disponibilité et concerne l’ensemble des professionnels de santé. C’est une rencontre interpersonnelle, un échange learn more de paroles, une circulation Adenylyl cyclase de sentiments et d’émotions qu’il faut savoir partager, écouter, et canaliser. Cette relation qui requiert de la

disponibilité, demande également une connaissance de soi et de ses propres limites ; elle se construit et s’élabore au fil du temps, dans un climat de confiance et de responsabilisation mutuelle par rapport au traitement proposé. Cette mission d’humanité exige une relation de vérité, d’authenticité du rapport à autrui. L’information donnée au malade (sur le diagnostic, le projet thérapeutique et l’évolution de la maladie) doit être claire, appropriée et loyale et nécessite d’avoir connaissance des limites de la médecine ; elle repose certes sur un « savoir-faire » scientifique spécifique, mais aussi et surtout sur un « savoir être » de tous les instants auprès de celui qui souffre. Il faut établir avec le patient, au fil du temps, au rythme des consultations successives, un climat de confiance de façon à faire émerger un projet thérapeutique aux objectifs partagés, tout en préservant l’autonomie du malade, en respectant ses choix de vie et en essayant de le rendre progressivement acteur dans la prise en charge de sa douleur. Il convient de travailler en coordination avec tous les acteurs de santé prenant en charge le patient.

The relative gene transfer was calculated by dividing the % value of each treatment by the % value for the standard. Here transconjugants serve as standard. Data were analyzed using Graph Pad InStat-3 and expressed as mean ± standard

deviation (SD) of three independent experiment. The continuous variables were tested with one-way analysis of variance (ANOVA) and Dunnett’s test. Values < 0.05 was considered statistically significant. Re-identification of all of the clinical isolates were done and found to be of A. baumannii, C. braakii, E. coli, P. aeruginosa and K. pneumoniae. A. baumannii and C. braakii were positive for both qnrA and qnrB gene, whereas E. coli, P. aeruginosa and K. pneumoniae were positive for qnrB gene and none of the clinical isolates harbored qnrS ( Fig. 1). As shown in the Table 1, Potentox emerged as the most active antibacterial against A. baumannii, P. aeruginosa, E. coli and K. pneumoniae with MIC values 8 μg/ml. EPZ5676 nmr The corresponding MIC for C. braakii was 16 μg/ml. The imipenem MIC values for A. baumannii and K. pneumoniae were 256 μg/ml each; 64 μg/ml for P. aeruginosa and C. braakii and 32 μg/ml for E. coli. The meropenem MIC values for A. baumannii, and K. pneumoniae were 128 μg/ml

each and 32 μg/ml for C. braakii and P. aeruginosa whereas 16 μg/ml for E. coli. For the other comparator drugs, the overall MIC values ranged from 32 to 1024 μg/ml. On the other hands, P. aeruginosa and K. pneumoniae found to be resistant to cefoperazone + sulbactam, amoxicillin plus clavulanic acid and levofloxacin; A. baumannii also showed resistant to amoxicillin plus clavulanic this website acid. There was a significant (p < 0.01) reduction in the MIC values of Potentox when compared

with the other comparator antibacterial agents ( Table 2). The zones of inhibition were calculated in millimeter for all strains and presented in the Table 3. Potentox was found to be sensitive against all clinical isolates as evident by zone of inhibition values, 23.5 ± 1.2, 20.8 ± 2.8, 25.8 ± 3.0, 27.2 ± 2.8, 23.2 ± 2.5 for A. baumannii, C. braakii, P. aeruginosa, E. coli and K. pneumoniae, respectively. Imipenem was found to be sensitive only against E. coli, already whereas meropenem was sensitive against P. aeruginosa and E. coli. Piperacillin plus tazobactam and cefoperazone plus sulbactam exhibited sensitivity toward C. braakii and E. coli. Cefepime was found to be sensitive only against C. braakii. Other tested drugs including amoxicillin plus clavulanic acid, moxifloxacin, levofloxacin and amikacin were observed to be resistant against all of the clinical isolates. The statistical analysis of AST values of Potentox vs other comparator drugs are shown in Table 4. Following conjugation, transconjugants were selected on MacConkey agar plates containing sodium azide and streptomycin. Analysis of transconjugants through PCR confirmed that transconjugants carrying the same gene as donor (Fig. 2).

The filtrate was used for the preliminary phytochemical analysis. The tests were performed according to methods described by Khandelwal (1998) and Kokate (2007). 12 and 13 TLC for various phytoconstituents was carried out as per methods described by Wagner and Bladt (1996).14 Albino Wistar rats, 8–12 weeks old, weighing in range of 120–180 g, was procured from Haffkine Institute, Parel. The animals were accommodated Selleckchem EPZ6438 in groups of five in polypropylene cages with stainless steel grill

top and a bedding of clean paddy husk was provided. The animals were maintained in air conditioned room with controlled temperature maintained in the range of 22–25 °C and alternating 12 h periods of light and dark cycle. The relative humidity was close to 60%. The animals were acclimatized to standard laboratory conditions prior to experimentation. The guidelines issued by Institutional Animal Ethics Committee of Ramnarain Ruia College, Mumbai, India with CPC SEA registration GSK1210151A no. CPC SEA/315, regarding the maintenance and dissection of small animals were strictly followed. Rats were administered a dose of 2000 mg/kg body weight for 14 days and were then examined for any signs of behavioural changes and mortality. All experiments were performed on female Albino Wistar rats (200–250 g)

obtained from the Haffkine Institute, Parel, Mumbai, Maharashtra, India. The animals were accommodated in groups of six in polypropylene

cages with stainless steel grill top and a bedding of clean paddy husk. Animals were maintained under a constant 12-h period of light and dark cycle and an environmental temperature of 22–25 °C. The Tryptophan synthase animals were acclimatized for 15 days before being used for the experiments. The guidelines issued by Institutional Animal Ethics Committee of Ramnarain Ruia College, Mumbai, India with CPC SEA registration no. CPC SEA/315, regarding the maintenance and dissection of small animals were strictly followed. The animals were fed on the standard pellet diet (Amrut Feed, Pune) and water was given ad libitum. The overnight fasted rats were made diabetic with streptozotocin (STZ) (Sigma, St Louis, MO; 60 mg/kg; intraperitoneally). The STZ was prepared freshly by dissolving it in Na-citrate buffer (0.01 M, pH 4.5) and maintained on ice prior to use; the injection volume was 0.2 ml. Diabetes was confirmed in the rats by measuring the fasting blood glucose concentration after 72 h of STZ administration. The rats with glucose level above 300 mg/dl were considered to be diabetic and were used in the experiment. Animals had free access to food and water after the STZ injection.