Lymphocytes were isolated from nasal-associated lymphoid tissues (NALT), nasal passages (NPs), head and neck lymph nodes (HNLNs), submaxillary glands (SMGs), spleens, small intestinal lamina propria (iLP), Peyer’s patches (PPs), lumbar lymph check details nodes (LLNs), sciatic lymph nodes (SLNs), and popliteal lymph nodes (PopLNs). HNLN, splenic, PP, LLN, SLN, and PopLN mononuclear cells were isolated by conventional methods using Dounce homogenization [26] and [27]. To isolate the mononuclear cells from NALT, NPs, SMGs, and iLP, the tissues were minced and digested using 300 units/ml of Clostridium histolyticum

Type IV collagenase (Worthington, Freehold, NJ) for 30 min at 37 °C in spinner flasks [26]. After incubation, the digestion mixtures were passed through Nitex mesh (FairviewFabrics, Hercules, CA) to remove undigested tissues. Mononuclear cells were separated by Percoll (Pharmacia, Uppsala, Sweden) density gradient centrifugation with cells interfacing between 40% and 60% Percoll. Greater than 95% viability was obtained for all lymphocytes isolated from

each tissue, as determined by trypan blue exclusion. On wk 14, sets of studies were terminated to collect NALT, NP, HNLN, SMG, splenic, iLP, PP, LLN, and PopLN mononuclear cells from the immunized mice. NALT, NP, HNLN, SMG, splenic, iLP, and PP mononuclear cells were used from i.n.-immunized mice, and NP, HNLN, splenic, iLP, LLN, and PopLN mononuclear cells were used from i.m.-immunized mice. Ag-specific Ab-forming cell (AFC) responses by the ELISPOT method were detected, using mixed Selleckchem LY294002 cellulose ester membrane-bottom microtiter plates (MultiScreen-HA; Millipore, Bedford, MA) by coating with 5 μg/ml F1- or V-Ag in sterile PBS, as previously described [27]. For total IgA or IgG AFC responses, wells were coated with 5 μg/ml goat anti-mouse IgA or IgG Abs (Southern Biotechnology Associates) in sterile PBS. On wk 7 or 14, groups of i.n.- or i.m.-immunized mice, respectively, were evaluated for cytokine responses to F1- and V-Ags. I.m.-immunized mice were boosted nasally with F1-Ag protein at 8 and 9 wks and with both DNA and nasally dosed with F1-Ag protein at 12 wks. From i.n.-immunized mice, HNLN,

splenic, and PP mononuclear cells were obtained, and HNLN, splenic, and peripheral lymph nodes (PLNs), containing Isotretinoin LLN, SLN, and PopLN mononuclear cells, were obtained from i.m.-immunized mice. Total mononuclear cells from each lymph tissue were resuspended in CM. Mononuclear cells were restimulated with 10 μg of recombinant F1-Ag, V-Ag, or with media as control in the presence of 10 U/ml human IL-2 (PeproTech) for 2 days at 37 °C in a humidified 5% CO2 incubator. Cells were washed and resuspended in CM, and then these stimulated lymphocytes were evaluated by IFN-γ-, IL-4-, IL-5-, IL-10-, and IL-13-specific ELISPOT assays, as described previously [24], [25] and [27]. To determine cytokine responses to F1- and V-Ags, on wk 7 or 14, groups of immunized i.n. or i.m. mice were used, respectively.

It can be scored from 0 to 3 for each response with a total possible score on the ranging check details from 0 to 84. Using this method, a total score of 23/24 is the threshold for the presence of distress. Alternatively the GHQ-28 can be scored with a binary method where Not at all, and No more than usual score 0, and Rather more than usual and Much more than usual score 1. Using this method any score above 4 indicates the presence of distress or ‘caseness’. Reliability and validity: Numerous studies have investigated reliability and validity of the GHQ-28 in various clinical populations. Test-retest reliability has been reported to be high (0.78 to 0 0.9) ( Robinson and Price 1982) and interrater and intrarater

reliability have both been shown to be excellent (Cronbach’s α 0.9–0.95) ( Failde and Ramos 2000). High internal consistency has also been reported ( Failde and Ramos 2000). The GHQ-28 correlates well with the Hospital Depression and Anxiety Scale (HADS) ( Sakakibara et al. 2009) and other measures of depression ( Robinson and Price 1982). The GHQ-28 was developed to be a screening tool and for this reason responsiveness in terms of Minimal Detectable Change (MDC) and Minimally Clinically Important

Difference (MCID) have not been established. Physiotherapists are becoming more aware of the need to screen for psychological and psychiatric co-morbidity in patients under their care. This may be to adapt or modify the physiotherapy approach to management or to institute referral to appropriate Selleck PD0325901 mental health care providers. The GHQ-28 is one of the most widely used and validated questionnaires to screen for emotional distress and possible psychiatric morbidity. It has been tested in numerous populations including people with stroke (Robinson and Price

1982), spinal cord injury (Sakakibara et al 2009), heart disease (Failde and Ramos 2000), and various musculoskeletal conditions including whiplash associated disorders (Sterling et al 2003) and occupational low back pain (Feyer et al 2000) amongst others. Thus for much clinicians there is a wealth of data with which to relate patient outcomes. It assesses the client’s current state and asks if that differs from his or her usual state. It is therefore sensitive to short-term distress or psychiatric disorders but not to long-standing attributes of the client. There are some disadvantages to use of the GHQ-28 in physiotherapy practice. First, the questionnaire is not freely available and must be purchased. Second, there is the potential for confusion over the different scoring methods, and this has implications for interpretation of scores derived from the questionnaire. There may also be some concern over the severe depression subscale which includes some confronting questions for the patient to answer. Other tools such as the HADS may be less confronting for physiotherapy use.

In particular, Ag-adsorbed NP enhanced T-cell proliferation responses in human PBMC (TT) and mouse splenocytes (HIV gp140). Also, gp140-adsorbed NP greatly enhanced serum IgG and IgA after systemic immunization and, more importantly, induced high levels of vaginal IgG and IgA after intranasal immunization. Solid lipid NP were prepared using a low pressure melt-emulsify-chill (MEC) process. A molten yellow carnauba (YC) wax (Koster Keunen, Watertown, CT) was dispersed into a hot

aqueous emulsifier solution under control shear and then cooled to yield a stable dispersion of solid lipid NP. For the preparation of fluorescence NP, the oil-soluble fluorescent dye Pyrromethene-567A (emission wavelength 546 nm, Exciton, Dayton, OH) was encapsulated in the NP. Cationic, anionic and non-ionic emulsifiers comprised 5-FU in vivo of long carbon chains were used to stabilize and also modify the surface charge of the NP. Particle size was determined by photon correlation spectroscopy using a Brookhaven BI90

Plus (Brookhaven Instruments, Holtsville, NY). The zeta (Z) potential (a measure of the surface electrical charge) of the NP and Ags was measured in 1 mM KCl by phase analysis light scattering using a Malvern Zetasizer NanoZS90 (Malvern Instruments, Malvern, UK). Particle morphology was analyzed by electron microscopy. Serial dilutions of the NP in nanopure water were dispensed in 400 nl medroxyprogesterone drops onto a silicon chip, and left to dry. Samples LY2157299 research buy were kept in the sputtering chamber at 5 × 102 mbar for about 4 h, and then sputter-coated with 15 nM gold. All images were taken at 20 kV, and at various magnifications using a Hitachi S3500N scanning electron microscope. NP colloidal stability was determined by storing 10% solid NP dispersions in glass vials at 5 °C and 25 °C. Particle size and

zeta potential were measured over a 12 month period as described above. For viscosity assessment, NP suspensions were stored in 125 ml plastic bottles for the length of the stability studies and the viscosity measured at different time points using a Brookfield viscometer LVT (Brookfield Engineering Labs, Middleboro, MA). Spindle #4 (low viscosity sample spindle) was placed directly in the sample, and speed setting 6 was used for all measurements. A clade C HIV-1 envelope clone p97CN54 was originally isolated from a Chinese patient [23] and was made available by H. Wolf and R. Wagner, University of Regensburg, Germany. Trimeric gp140 (gp120 plus the external domain (ED) of gp41), designated CN54 gp140, was produced as a recombinant product in CHO cells and manufactured to GMP specification by Polymun Scientific, Vienna, Austria. Bovine serum albumin (BSA) and TT were obtained from Sigma–Aldrich, Ayrshire, UK and Statens Serum Institute, Denmark, Copenhagen, respectively.

, 2014). In conjunction with findings in animal models, these results are consistent with the hypothesis that stress-associated changes in connectivity in large-scale brain networks are PLX4032 an important feature of depression and other stress-related neuropsychiatric disorders, and that resilience and vulnerability may be determined

in part by individual differences in the capacity for plasticity within these circuits. Understanding the mechanisms by which stress alters connectivity in vulnerable circuits may reveal new avenues for treatment. Undoubtedly, many factors are involved, and some of them have been reviewed elsewhere (De Kloet et al., 1998a, McEwen, 2000, De Kloet et al., 2005b, Arnsten,

2009, Joëls and Baram, 2009 and Chen et al., 2010). Here we focus on a factor that has received relatively little attention, namely, endogenous glucocorticoid oscillations and their role in regulating synaptic plasticity. Glucocorticoids are hormones that are released from the adrenal gland in response to signals originating in the pituitary and hypothalamus, which receives projections from distinct circuits for detecting physiological and psychosocial stressors (Herman and Cullinan, 1997 and Ulrich-Lai and Herman, 2009) (Fig. 2a). In the short term, glucocorticoids serve to mobilize energy resources and facilitate sympathetic nervous system responses to maintain homeostasis and adapt click here to stress. In the long term, however, prolonged exposure to glucocorticoids in chronic stress states can have maladaptive effects, mediated in part by disruptions in negative feedback mechanisms (McEwen, 1998 and McEwen, 2003). Glucocorticoid activity also oscillates with diurnal activity rhythms, independent of external stressors (Fig. 2b): glucocorticoid secretion tends to peak in the early morning in diurnal animals (early Thymidine kinase evening in nocturnal animals), remains relatively elevated for most of the active period of the animal’s

day, and becomes relatively suppressed for most of the night. In addition, recent reports (Stavreva et al., 2009a and Lightman and Conway-Campbell, 2010) have shown that an ultradian oscillation with a period of 1–2 h is superimposed on this circadian rhythm and has equally important consequences for glucocorticoid signaling (reviewed below). In previous fixed tissue studies, stress and glucocorticoid effects on dendritic arborization and spine density took weeks to develop (Magariños et al., 1996, Wellman, 2001, Vyas et al., 2002, Radley et al., 2004 and Radley et al., 2006), which would imply that glucocorticoid oscillations occurring on a timescale of minutes to hours were unlikely to play a direct role in these changes. However, recent studies indicate that glucocorticoids and related signaling molecules can have much more rapid effects on dendritic spines than were previously suspected.

The concentrations find more of sodium and potassium ions were determined by the method of.11 The data obtained from the laboratory results of the tests were

subjected to One Way Analysis of Variance (ANOVA). Significant differences were observed at p ≤ 0.05. The results were expressed as means of five replicates ± standard deviations (SD). This analysis was done using the computer software known as Statistical Package for Social Sciences (SPSS), version 16. The qualitative phytochemical analyses showed, the presence of saponins in very high concentration in both fractions of the chloroform–methanol extract of the leaves of P. americana ( Table 1). Flavonoids were found to be present in very high concentration selleck and moderately high concentration in the chloroform and the methanol fractions respectively. Tannins, terpenoids and steroids were found to be present in moderately high concentrations in both fractions

as shown in Table 1. The phytochemical constituents of the chloroform and the methanol fractions of the chloroform–methanol extract of the leaves of P. americana are summarised in Table 2. The chloroform fraction of the extract contained higher percentages of alkaloids (2.67 ± 0.13%), flavonoids (3.20 ± 0.17%) and steroids (1.36 ± 0.04%) than the methanol fraction while the methanol fraction contained higher percentages of saponins (2.23 ± 0.09%) and tannins (2.73 ± 0.13%) than the chloroform fraction ( Table 2). The charcoal meal (gastro-intestinal motility) test was used to determine the propulsive movement along the gastro-intestinal tract (GIT) of rats. As shown in Fig. 1, the methanol and the chloroform fractions of the extract at the tested doses (100 and 200 mg/kg body weight of each) significantly (p < 0.05) decreased the percentage distance travelled by the charcoal meal along the gastro-intestinal tract of rats in groups 4, 5, 6 and 7 when compared to those the value obtained for rats in the charcoal meal-treated control

group (group 2). The observed effects were dose-dependent with percentage distance travelled by charcoal meal as 62.25 ± 4.57, 57.25 ± 1.50, 58.25 ± 2.22 and 35.25 ± 2.36 for rats in the 100 and 200 mg/kg body weight of the methanol fraction-treated groups (groups 4 and 5), 100 and 200 mg/kg body weight of the chloroform fraction-treated groups (groups 6 and 7) respectively when compared to the value (70.25 ± 3.30) obtained for rats in the charcoal meal-treated control group (group 2). The effects of the methanol and the chloroform fractions of the extract at the tested doses were comparable to that of the standard anti-diarrhoeal agent (hyoscine butylbromide) as shown in Fig. 1. Result of the intestinal fluid sodium ion concentration test as shown in Fig. 2, shows that the rats of the castor oil-treated control group (group 2) had significantly (p < 0.05) increased intestinal fluid sodium ion concentration (227.00 ± 3.46) when compared to the value (192.75 ± 11.

These properties are very promising devices for gene therapy of new age (Cytoplasmic Gene Therapy) because of its genotoxicity-free nature. Further, it is non-pathogenic for humans. Since Sendai virus is a murine parainfluenza virus (PIV) with certain homologies to human PIV, it was tested

as xenotropic vaccine in African Green monkeys and humans without any significant adverse reactions [34] and [35]. Recombinant SeV vector carrying human PIV was also tested in rats [36] and [37]. Further, recombinant SeV vaccine for human immunodeficiency virus (HIV) infection is going to be tested in humans (http://www.dnavec.co.jp/en/index.html). Thus, safety of Sendai virus vector is gradually established. We inserted mouse IL-10 cDNA to construct rSeV-Aβ with the aim of helping antibodies productions and suppressing Th1 type T cell activations. Nasal administration of rSeV-Aβ without IL-10 had less effect to remove Rapamycin Aβ depositions (data not shown). Recently, soluble Aβ oligomers, but not fibrils nor GDC-0199 concentration monomers, have been considered responsible for cognitive dysfunction prior to the formation of Aβ plaques [22], and Aβ*56, a 56-kDa soluble Aβ dodecamer was found responsible in Tg2576 mice [29]. Our nasal vaccine efficiently reduced not only senile plaque amyloid but also the contents of Aβ*56 oligomer without changing sAPPα and improved cognitive dysfunction in water

maze, Y-maze and contextual fear test which could evaluate hippocampus-related cognition. Thus, our vaccine, if applicable, can be given at the stage of mild cognitive impairment or earlier. Aβ is released from presynaptic sites and deposited in and extracellular plaques [38], and APP and synaptophysin are co-localized at the growth cones of developing neurons in culture [39]. These reports have indicated that Aβ deposition plays an important role in degeneration of presynaptic structures. In addition, it is reported that Aβ oligomers

directly disrupt synaptic structures [40]. In our study, synaptophysin staining showed amelioration of presynaptic degeneration following our nasal vaccine at 24 months old, suggesting prevention of synaptic degeneration or repair of synaptic structures after removal of Aβ deposits including Aβ oligomers. Our next plan is to see whether Tg2576 mice show improvement of cognitive functions by eliminating senile plaque amyloid even at 24 months old. In conclusion, a new vaccine using Sendai virus vector with Aβ and IL-10 cDNA was developed. A nasal administration of this vaccine reduced amyloid burden including Aβ oligomers significantly in AD mice and improved cognitive functions without causing side effects such as brain inflammation. This vaccine can be used to treat and prevent Alzheimer disease. Authors are grateful to Dr. Y. Noda at Meijo University, Dr. T. Nagai at Nagoya University, and Dr. M.

Marine natural products provide a novel and rich source of chemical diversity that can contribute to design and development of new and potentially useful anticancer agents. All Selumetinib chemical structure authors have none to declare. “
“Cancer is a second leading cause of mortality and morbidity all over the world.1 Plant derived anticancer agents are widely used for the treatment of cancer. In our continued interest in search of anticancer agents from Indian medicinal plants, we have investigated anticancer potential of Cuscuta reflexa Roxb. (Family: Convolvulaceae, Amervel in Hindi). It is used indigenously in Indian system

of medicine in the remedy for various ailments. Various parts of the plant are used by tribes for treating Screening Library molecular weight ailments like fits, melancholy and insanity 2 and to control fertility. 3 It is also used externally against itch and internally in fevers, ‘retention of wind’ and induration of the liver 4, 5 and 6 in the indigenous system of medicines. Preliminary pharmacological studies are reported in literature. 7 The plant is reported for α-glucosidase

inhibitory, 8 free radical scavenging, 9 antibacterial, 10 psychopharmacological, 11 antisteroidogenic 12 and hair-growth promoting 13 activities. The chemical compounds isolated from the plants are mainly flavonoids. 14, 15 and 16 The present work was undertaken to evaluate the antiproliferative potential of Cuscuta reflexa Roxb. RPMI-1640, Dulbecco’s minimum essential medium (DMEM), Fetal calf serum (FCS), trypsin, gentamycin, Terminal deoxynucleotidyl transferase penicillin, ethylene diaminetetraacetic acid (EDTA), 5-Flurouracil, dimethyl sulphoxide, sulforhodamine-B were purchased from Sigma Chemical Co; USA. All other chemicals were

of high purity and obtained locally. Whole plant of Cuscuta reflexa was collected locally in the month of December and was authenticated at source by the taxonomist of the institute. A voucher specimen has been deposited at the herbarium of the Institute vide IIIM collection No.17148, Acc. No.17719. The authenticated and freshly collected whole plant was chopped and dried under shade. Three extracts of the plant material were made with 95% alcohol, alcohol-water (1:1) and water using repeated solvent extraction procedure. 17 Dried powdered plant material (1 kg) was percolated in 95% ethanol (5 L) at ambient temperature for 16 h. The solvent was decanted and the process was repeated four times. The pooled solvent was evaporated under reduced pressure to yield alcoholic extract (160 g). Similarly, hydro-alcoholic extract was prepared. The dried plant material (200 g) was soaked in alcohol-water (1:1, 1 L) and the extract obtained was 72 g. The dried powdered plant material (200 g) was heated with distilled water (1.

The electropherograms obtained were analyzed using the sequencing analysis software (Sequence Navigator, version 1.01, Applied Biosystems). The nt and deduced aa sequences were compared with sequences available in the NCBI (National Center for Biotechnology Information) GenBank database using the BLAST (Basic Local Alignment Search Tool) program. Phylogenetic and molecular buy Y-27632 evolutionary analyses were conducted using MEGA version 4.0 [36]. Dendrograms constructed were confirmed by two different methods,

neighbor joining and maximum parsimony. The data were analyzed using Epi Info 2002 and Stata 10.0. Chi square and Mann Whitney U tests were performed to determine the significance of differences observed between groups. Partial nucleotide Androgen Receptor Antagonist sequences of VP1, VP2, VP3, VP4, VP6, VP7, NSP1, NSP2, NSP3, NSP4 and NSP5 of the G10P[15] strains were submitted to the GenBank database and their accession numbers are HQ660637, HQ660638, HQ660639, FJ798615, FJ798616, FJ798617, HQ660640, HQ660641, HQ660642, FJ798618, HQ660643 respectively. The median (interquartile range [IQR]) age of the 394 children enrolled in the study was 10 (7) months, with >90% of children less than 2 years of age. The median Vesikari score of diarrheal severity was 11.0 and the children required

admission for a mean duration of 2.8 days. Of 394 children screened, we found that 158 children were infected with rotavirus (40%). The common G types identified in order of frequency were G1 (47/158, 29.7%), G2 (43/158, 27.2%), G9 (22/158, 13.9%), G10 (2/158, 1.2%), G12 (1/158, 0.6%) and mixed infections (27/158, 17.8%). The common P types were P[4] accounting for 57/158 (36%) samples, P[8] 57/158 (36%), P[11] 3/158 (1.8%) and P[6] 2/158 (1.2%). Mixed infections with varied P types were seen in 5 (3.2%). G typing alone was possible in 23 samples Thalidomide (14.4%), only P typing in 5 samples (3.6%) and 11 samples were completely untypeable (6.9%). The common G:P combinations seen

in children were, G2P[4] in 39/158 (24.6%) samples, G1P[8] in 29/158 (18.3%) samples, G9P[8] in 21/158 (13.2%) samples, G1P[4] in 4/158 (2.5%) samples and G10P[11] in 1/158 sample (0.6%) (Fig. 1a). We collected total of 627 samples from animals with diarrhea, including 589 cows (25 were calves), 2 buffaloes, 11 bullocks and 25 goats (11 were kids). The mean duration of diarrhea was 4.5 days for adult animals, 4 days for calves and 3 days for goat kids. Out of 627 animals we found 35 (1 bullock, 2 goats, 32 cows) infected with rotavirus (5.5%). The common G types identified in order of frequency were G6 (17/35, 48.5%), G2 (10/35, 28%), G10 (4/35, 11%), G8 (2/35, 5.7%) and mixed infections (2/35, 5.7%).

A portion of the work described herein was carried out by Jennifer Kasper in partial fulfilment of the requirements for a biological doctoral degree at the Johannes Gutenberg University, Mainz, Germany. The authors wish to thank Ms. Elke Hübsch and Ms Michaela Moisch for their excellent assistance with the cell culture and immunocytochemical

studies. This study was supported by the DFG priority program SPP 1313 within the Cluster BIONEERS and also by the European Union, FP6 Project NanoBioPharmaceutics. “
“The applications of microparticles and nanoparticles Ibrutinib ic50 as delivery vehicles or therapeutic entities are widely described in the literature. Their combination, for example, as nanoparticle-in-microparticle (NIM) systems, offers the possibility of dual or multiple functionalities within a formulation. For example, multiple release profiles (burst release from outer particles selleck chemical and sustained release from internal components) and/or combinations of features allowing site

specificity, in vivo protection, cellular interactions, imaging capabilities and embolisation can all be envisaged. In recent examples, Veiseh et al. proposed multifunctional delivery systems comprising both imaging and therapeutic agents, in addition to a functionalised surface to enhance specific cell interactions [1]. Pouponneau et al. produced a microparticle system that encapsulated magnetic Ketanserin nanoparticles and showed that under the influence of a magnetic field, the particles could be steered in vitro [2]. Another example includes theophylline-loaded NIM suitable for asthmatic treatment in which Jelvehgari et al. utilised the outer microparticle as a means to reduce burst release [3]. Various methods have been proposed for the preparation of NIM systems. Spray drying techniques have been used to produce NIMs for aerosols [4], [5], [6] and [7], oral [8] and [9] and intravitreal

formulations [10]. Other methods include supercritical fluid techniques [11], [12] and [13]. There is, however, little information on how NIMs can be produced using the standard emulsion techniques that are widely and conveniently used in the preparation of particles for drug delivery research. Such methods for preparing single-component particles (i.e. microparticles or nanoparticles alone) are renowned for their application to both hydrophilic or hydrophobic drugs and a variety of polymer systems [14]. Additionally, through modification of process parameters, characteristics such as particle size distribution and morphology can be readily altered. While work such as Jelvehgari et al. [3] provides methodology for NIM formation, there is little convincing information in the drug delivery literature on the internal structure of NIMs or the distribution of nanoparticles therein.

Range was established with five replicate readings of each concentration. Precision of the method was determined in the terms of intra-day and inter-day variation (%RSD). Intra-day precision (%RSD) was assessed by analysing standard drug solutions within the calibration range, three times on the same day. %RSD was found to be 0.30–1.14 for TDF and 0.51–1.37 for ETB. Inter-day precision (%RSD) was assessed by analysing drug solutions within the calibration range on three different days over a period of a week. Vismodegib concentration %RSD was found to be for TDF and 0.57–1.08 for ETB. This indicates that adequate preciseness of the method. Detection limit and quantification limit was calculated by the method as described in Section 2.4.2. The LOQ

and LOD for DAPT TDF were 13.99 ng and 42.40 ng. For ETB, LOQ and LOD were found to be 7.37 ng and 22.32 ng, respectively. This indicates that adequate sensitivity of the method. To the preanalysed sample a known amount of standard solution of pure drug (TDF and ETB) was over spotted at three different levels. These solutions were subjected to re-analysis by the proposed method and results of the same are shown in Table 2. The standard deviation of peak areas was calculated for each parameter and %R.S.D. was found 0.65–2.00. The low %R.S.D. indicates robustness of the method. The ruggedness of the proposed method was evaluated

by two different analysts. The results for TDF and ETB 3-mercaptopyruvate sulfurtransferase were found to be 99.78%, 99.50% and 100.64%, 100.28%, respectively. Repeatability of sample application was assessed by spotting (300 ng/spot) of drug solution seven times on a TLC, followed by development of plate and recording the peak area for seven spots. The %R.S.D. for peak

area values of TDF and ETB was found to be 1.21 and 0.57, respectively. The summery of validation parameters were listed in Table 3. The chromatogram of samples degraded with acid, base, hydrogen peroxide and light showed well separated spots of pure TDF and ETB as well as some additional peaks at different Rf values. The number of degradation product with their Rf values, content of TDF and ETB remained, and percentage recovery were calculated and listed in Table 4. The proposed HPTLC method provides simple, accurate and reproducible quantitative analysis for simultaneous determination of TDF and ETB in tablets. The method was validated as per ICH guidelines. All authors have none to declare. The authors are thankful to R.C. Patel College of Pharmacy for providing necessary facilities. “
“Diabetes associated complications have become a public health problem of considerable magnitude, because of huge premature morbidity and mortality associated with diabetes. Hyperglycemia inherent to diabetes patients accelerates accumulation of advanced glycation end-products (AGEs). Formation of AGEs is a slow non-enzymatic glycation process when reducing sugar reacts with proteins through a series of irreversible reaction and rearrangement.