40 reported that TEE values obtained from HR monitoring were 190 kcal/day higher as compared to DLW. In contrast, Van den Berg-Emons and colleagues41 found that DLW gave higher TEE values than HR by 17 kcal/day. In the current study, we found that TEE estimated by HR analysis was similar to that assessed

by DLW in ordinary males and females. The average difference was only 9 kcal/day, which was better than the above-mentioned studies. Thus, our results indicate that HR analysis using Suunto’s software (MoveSense HRAnalyzer 2011a, RC1) can be applied for TEE estimation in a free-living ordinary population at the group level. The REE is the amount of energy expended by the metabolically active components of the Ibrutinib mw body at rest. FFM accounts for about 65%–90% of the individual variance in REE,27 and the REE accounts for about 60%–75% of the TEE.6 The knowledge of the source of the REE and its relationship with the TEE has been used as a basis for establishing effective weight management programs.42 Substantial efforts have been made to develop age- and gender-specific28, 43, 44 and 45 or body composition-specific27 and 46 equations to estimate the REE. A Swiss study group47 compared

five equations; the Harris–Benedict, Mifflin–St Jeor, Owen, World Health Organization and Lührmann methods, to indirect calorimetry, and found that the mean differences varied between −41 and 53 kcal/day in the elderly. We found mean differences of 25, 28, and 73 kcal/day in middle-aged women, men and young women, respectively, indicating that the Harris–Benedict equation overestimated the REE, NVP-BGJ398 mouse especially in the younger female population, as compared to indirect calorimetric GEA. Thalidomide Few studies have validated the Cunningham equation used by BIA against indirect calorimetry.10 and 48 We found that there were no significant differences in the REE estimates between GEA and the Cunningham equation used by BIA (InBody 720) among middle-aged

men and women. However, the Cunningham equation significantly underestimated the REE in 19-year-old young women. This indicates that the relationship between FFM and the REE is probably age-specific, and the predictive equations derived from adults may not be applicable to younger people. The major limitation of our study was that the TEE estimation using HR analysis was based on a 24-h recording, whereas the TEE derived from DLW reflects the average daily energy expenditure over 14 days. This is one of the main causes underlying the differences between TEE estimates from HR analysis vs. DLW and the large individual variation. However, the variance in TEE estimation between the DLW and HR methods found in our study has also been reported in other studies. 11, 16, 22, 40 and 49 The HR monitoring has an advantage in monitoring day-to-day energy expenditure, which is important for most practical purposes.

, 2009, Lee et al., 2010, Moon et al., 2006 and Moon et al., 2009). Analysis of Gr-GAL4 drivers has shown that Gr5a is expressed in sugar-sensitive neurons in each sensillum, while Gr66a is expressed in a distinct population

of ∼20 neurons that responds to a number of bitter compounds and that mediates aversion ( Chyb et al., 2003, Marella et al., 2006, Thorne et al., 2004 and Wang et al., 2004). Two Gr5a-related genes map to Gr5a-expressing neurons, while a number of other Gr genes appear to be expressed in subsets of Gr66a-expressing neurons ( Dahanukar et al., 2007, Lee et al., 2009, Moon et al., 2009, Thorne and Amrein, 2008, Thorne et al., 2004 and Wang et al., 2004). The sensilla associated with these subsets have not been identified in most

cases, however, and expression of the great majority of Gr genes has not been examined. Historically, a critical question check details in the field has been whether all taste sensilla are functionally equivalent (Hiroi PI3K inhibitor et al., 2002, Marella et al., 2006, Thorne et al., 2004 and Wang et al., 2004). Previous physiological analysis of the labellum revealed that three sensilla, L7, L8, and L9 (Figure 1A), were similar in their responses to all of 50 tested compounds, mostly sugars (Dahanukar et al., 2007). A study of 21 sensilla and four sugars showed that all sensilla responded to all tested sugars, with some quantitative differences among sensilla of different morphology (Hiroi et al., 2002). A survey of a few bitter compounds revealed that none of the longer sensilla on the labellum responded, while all of the shorter hairs that were tested gave indistinguishable responses (Hiroi et al., 2004). An imaging study found that different subpopulations of bitter cells responded to most bitter compounds tested; striking differences in response profiles were not observed (Marella GPX6 et al., 2006). Based on these studies, it has been suggested that bitter-sensitive neurons of the labellum may generally recognize the same bitter

compounds (Cobb et al., 2009 and Marella et al., 2006). A similar model emphasizing functional homogeneity is often cited in mammals, in which multiple bitter receptors are coexpressed and taste receptor cells respond to a broad range of bitter compounds (Adler et al., 2000, Mueller et al., 2005 and Yarmolinsky et al., 2009). However, a systematic analysis of the responses of the labellar taste sensilla to bitter compounds, such as those carried out with Drosophila olfactory sensilla and odorants ( de Bruyne et al., 2001), has not been performed. Because of the limited scope of the extant studies, the basic principles of functional organization that underlie bitter coding in the fly remain unclear. Here we investigate basic principles of bitter coding through a systematic behavioral, physiological, and molecular analysis.

Nuclei were visualized by incubating click here for 10 min with 0.1 μg/ml 4,6-diamidino-2-phenylindole

(DAPI; Sigma-Aldrich, St. Louis, MO, USA) in PBS and F-actin filaments by incubation in Texas Red-labeled phalloidin (5 U/ml; Invitrogen, Grand Island, NY, USA). Fluorescent secondary antibodies were used according to the manufacturer’s protocol (Jackson ImmunoResearch, West Grove, PA, USA; Southern Biotechnology, Birmingham, AL, USA; Invitrogen), and sections were analyzed using Olympus or Leica laser-scanning microscopes. Pregnant mice were operated on as approved by the Government of Upper Bavaria under license number 55.2-1-54-2531-144/07 and were anaesthetized by intraperitoneal injection of saline solution containing Fentanyl (0.05 mg/kg), Midazolam (5 mg/kg), and Medetomidine (0.5 mg/kg;Betäubungsmittel license number: 4518395), and E13/E14 embryos were electroporated as described before (Saito, 2006). pCIG2 containing a GFP or CreGFP, RhoA∗GFP and Gap43GFP plasmids were mixed with Fast Green (2.5 mg/μl; Sigma) and injected at the concentration of 1μg/μl. Anesthesia was terminated S3I-201 manufacturer by Buprenorphine (0.1 mg/kg), Atipamezol (2.5 mg/kg), and

Flumazenil (0.5 mg/kg). Embryos were fixed in 4% paraformaldehyde (PFA), and vibratome sections of 100 μm were analyzed using Olympus laser-scanning microscopes. Cortical embryonic cells were dissociated and incubated in trypsin for 15 min with green cell tracker (Invitrogen C7025), and 70.000 cells/μl were resuspended in Dulbecco’s modified eagle medium, and 1 μl was injected into the ventricle of E13 or E14 embryos. Mice were anaesthetized as described previously. Embryos/pups were fixed in 4% PFA, and vibratome sections of 100 μm were analyzed using Olympus laser-scanning microscopes. GFP, Ph3, Tbr1, Ctip2, Cux1, and GAD67+ cells were quantified by counting all positive cells in a radial stripe comprising all cortical layers. Quantifications are given as the mean ± SEM; statistical significance was tested

with the unpaired student’s t test or unequal variance t test. c-Fos and Egr-1+ cells were quantified using a NeuroLucida device and StereoInvestigator software (MBF Bioscience, Magdeburg, Germany). Cells were counted in a vertical stripe (layers II–VI). At least five sections for each experimental next animal were examined. Cortices from embryonic brains were lysed in RIPA buffer containing protease and phosphatase inhibitors (Roche, Madison, WI, USA), and 20 μg of total protein were separated by 10% SDS-PAGE and transferred to PVDF membranes (Biorad, Berkley, CA, USA), which were incubated with primary antibodies followed by horseradish peroxidase-labeled secondary antibodies (1:25000; Amersham, Little Chalfont, UK) detected by ECL Western Blotting Detection (Millipore, Billerica, MA, USA). Quantification of bands was performed using ImageJ software. Primary antibodies are listed in Table S1. The G-actin and F-actin fractions were separated by centrifugation (Posern et al., 2002).

“
“In the original paper, we reported behavioural and ERP measures of response inhibition in the stop-signal task in young female heavy drinkers. We have recently discovered an error in the processing of the probability of inhibition for the earliest stop-signal delay, which had carry-on effects to calculations of stop-signal reaction time (SSRT). This results in minor changes to the significance level of analyses involving the probability selleck chemical of inhibition and SSRT, but does not change the interpretation of these results, and does not affect the ERP results.

The correct results for Table 1, first paragraph of the Results section, and an updated Fig. 1 appear below. Table 1. Group characteristics and performance data for controls (n = 17) and heavy drinkers (n = 13). All values are represented as mean ± SD. F statistics have (1, 28) degrees of freedom. Fig. 1a shows that the probability of successfully inhibiting a response on stop-signal trials decreased with increasing stop-signal delay (F = 349.357, p = .000), and heavy-drinkers were less able to inhibit EPZ-6438 mouse responses at every delay compared to controls (approaching significance; see Table 1 for group statistics). There was no difference in the slope of the inhibition functions between groups (F < 1). The heavy drinking group showed a significantly longer SSRT than the control group,

indicating deficient inhibition. Mean RT and accuracy to Go trials were not significantly different between groups. We also observed a strong positive correlation between scores on also the AUDIT and SSRT, such that more hazardous and harmful drinking was associated with a longer SSRT (p = .000, see Fig. 1b). If groups were created based on AUDIT scores, hazardous and harmful drinkers (score ≥ 8) showed longer SSRT (263.90 ± 37.61 ms, mean ± SD) than low-risk drinkers (score ≤ 7, 224.90 ± 33.61 ms; F = 8.82, p = .006). “
“The article to follow was accidently omitted from the March 2011 issue. We apologize for the error. “
“Attention deficit hyperactivity disorder (ADHD) is a

childhood developmental disorder characterized by symptoms of inattention, hyperactivity and impulsivity. In children with ADHD, a wide range of impairments in cognitive functions are found, particularly regarding executive functions (i.e., response inhibition, working memory, planning, selective and divided attention, set-shifting, and time processing; O’Brien et al., 2010, Pasini et al., 2007, Valko et al., 2010 and Willcutt et al., 2005). While ADHD symptoms often wane in adulthood, these symptoms may persist in some patients. Studies in adult ADHD patients reported on deficits in working memory (Finke et al., 2011 and Marx et al., 2011), reward and emotional processing (Wilbertz et al., 2012, Marx et al., 2011 and Ibáñez et al., 2011), time processing (Valko et al., 2010), and inhibitory control (Bramham et al., 2012, Cummins et al., 2011 and Wilbertz et al., 2012).

Within the piriform cortex, layer 2/3 pyramidal cells receive direct sensory input from M/T cells on their apical dendrites. Whereas olfactory information is encoded as a spatial map of activated M/T cells in the olfactory bulb,

odor representations in layer 2/3 of the piriform cortex are distributed Selleckchem BKM120 among spatially dispersed cell ensembles and lack stereotypy (Illig and Haberly, 2003, Rennaker et al., 2007 and Stettler and Axel, 2009). The mechanisms governing this transformation from a spatially segregated representation in the olfactory bulb to one that is highly distributed and nonstereotyped in the cortex are not well understood. Individual pyramidal cells in the piriform cortex are thought to receive converging input from M/T cells belonging to different glomeruli (Apicella et al., 2010, Davison and Ehlers, 2011, Miyamichi et al., 2011 and Wilson, 2001), and M/T cell axons from individual glomeruli project diffusely throughout the piriform cortex without obvious spatial patterning (Ghosh et al., 2011 and Sosulski et al., 2011). Although it is tempting to account for cortical odor responses PI3K inhibitor entirely by the convergence and divergence of direct olfactory bulb inputs, the dendrites of the piriform cortex pyramidal cells also receive extensive intracortical associational (ASSN) connections from excitatory neurons

within the piriform cortex and other cortical regions (Haberly, 2001, Haberly and Price, 1978 and Johnson et al., 2000). Although much effort has focused on elucidating how olfactory bulb afferent sensory inputs shape cortical odor representations, the contribution of intracortical excitatory circuits

to odor responses has been largely unexplored. In this study, we examine the relative contributions of sensory afferent input and intracortical connections to odor-driven excitatory synaptic transmission in the anterior piriform cortex (APC). We take advantage of the only differential expression of presynaptic GABAB receptors in APC to selectively silence intracortical synapses while leaving afferent sensory fibers unaffected. We show that intracortical connections in APC underlie the strength of odor-evoked excitatory synaptic transmission and expand the range of odors over which pyramidal cells can respond. Our results indicate that intracortical ASSN circuits make a major contribution to odor-evoked excitation, suggesting that odor representations in the piriform cortex cannot simply be accounted for by the convergence and divergence of M/T cell inputs. GABAB receptors are expressed on nerve terminals, and activation of presynaptic GABAB receptors causes a potent inhibition of neurotransmitter release from both pyramidal cells and local interneurons throughout the cortex (Bowery, 1993).

In the LC of line 3.1, 97% ± 1% of neurons that expressed ChR2-YFP also expressed TH, while 72% ± 6% of neurons that expressed TH also expressed ChR2-YFP (n = 122 for VTA, n = 86 for SN, n = 63 for LC, where n refers to counted cells; Figure 1A). To further characterize these lines, we quantified the copy number of Cre in Th::Cre rats with digital PCR ( Supplemental Experimental Procedures). Across multiple

sublines (Th::Cre 3.1, 3.5, and 4.4), we observed a single copy number of Cre in the genome ( Table S1). We performed a systematic in vitro electrophysiological study of the cellular and optogenetic properties of ChR2-YFP-expressing VTA neurons in Th::Cre+ rats, along with a comparison http://www.selleckchem.com/products/byl719.html of the same properties of neurons in Th::Cre+ littermates injected with a control virus that expressed only

YFP. Figure 2A shows a sample trace from a ChR2-expressing Th::Cre neuron in response to current injection steps, demonstrating the classical “sag” response induced by the hyperpolarizing pulse, the result of a hyperpolarization-activated cation current (Ih) that is present in many TH+ VTA neurons ( Margolis et al., 2006, Lammel et al., 2008, Lammel et al., 2011 and Neuhoff et al., 2002). Given that the VTA TH+ neurons are heterogeneous and do not all express a prominent Ih ( Lammel et al., 2008 and Lammel et al., 2011), in addition to analyzing light responses and intrinsic properties of neurons with a prominent Ih current (Ih/large neurons), we have also included in this http://www.selleckchem.com/products/s-gsk1349572.html analysis cells without a prominent Ih (Ih/small neurons), in either case comparing the properties of neurons that express ChR2-YFP to neurons

that express YFP only. Continuous blue light elicited large inward currents (peak photocurrent: −2950 ± 1574 pA for Ih/large neurons, steady-state photocurrent: −756.5 ± 225.5 pA; n = 7 Ih/large neurons, Figure 2B), and optical stimulation trains produced neural responses that were similar to those evoked by isothipendyl electrical stimulation in both ChR2-expressing or YFP-only-expressing neurons (Figure 2C for pulse trains, Figure S2D for individual waveforms); notably, the amplitude of both optically and electrically evoked spike trains attenuated during the course of the pulse train. Th::Cre ChR2-expressing neurons reliably responded to light-induced spike trains over a range of frequencies from 5 to 40 Hz ( Figure 2D for Ih/large neurons, Figure S2B for Ih/small neurons). Multiple spikes in response to a single light pulse were never observed during the presentation of pulse trains under these expression, illumination, and opsin (ChR2) conditions. We also confirmed that light stimulation at various frequencies (5 to 40 Hz) failed to evoke neural responses in YFP-only-expressing neurons ( Figure S2C).

Lastly, while the mutant mouse in the current study and the hearing loss described in patients with DFNA25 are both due to mutations in the gene coding for VGLUT3,

the comparison may not be straightforward. First, it is not certain that the missense mutation described in SLC17A8 is the cause of the hearing loss seen in DFNA25, though a strong correlation was observed (Ruel et al., 2008). Second, the null mutation studied in these experiments would represent a much more severe phenotype than the missense mutation described as potentially causative for DNFA25. Thus, whether this technique could ultimately be beneficial to patients with DFNA25 remains unclear. Despite these differences, as our study documents restoration of normal ABR levels in such a null mutant model, it nonetheless represents an important initial step for the potential treatment PD0332991 manufacturer of inherited deafness. VGLUT3 null mutant mice were generated as described in a C57 (Seal et al., 2008) strain Ponatinib chemical structure then backcrossed with FVB mice (less than seven generations) to obtain a homogeneous genetic background. P1–P12 mice were used for AAV1-VGLUT3 delivery. All procedures and animal handling complied with NIH ethics guidelines

and approved protocol requirements at the University of California, San Francisco (IACUC). All surgical procedures were done in a clean, dedicated space. Instruments were thoroughly cleaned with 70% ETOH and autoclaved prior surgery. Surgery was carried out under a Leica MZ95 dissecting scope and animals were situated with neck extended over solid support. Mice were anesthetized by intraperitoneal injection of a mixture of aminophylline ketamine hydrochloride (Ketaset, 100 mg/kg), xylazine hydrochloride (Xyla-ject, 10 mg/kg), and acepromazine (2 mg/kg) and boosted with one-fifth the original dose

as required. Depth of anesthesia was continuously checked by deep tissue response to toe pinch. Body temperature was maintained with a heating pad and monitored with a rectal probe throughout procedures. Preoperatively and every 24 hr postoperatively, animals were given subcutaneous carprofen analgesia (2 mg/kg) to manage inflammation and pain. Animals were closely monitored for signs of distress and abnormal weight loss postoperatively. Mouse VGLUT3 cDNA was subcloned into the multiple cloning site of vector AM/CBA-WPRE-BGH (kindly provided by R. Palmiter). Human embryonic kidney 293 cells were cotransfected with three plasmids—AAV-mVGLUT3 plasmid, appropriate helper plasmid-encoding rep and AAV1 cap genes, and adenoviral helper pF Δ6—using standard CaPO4 transfection. Cells were harvested 60 hr after transfection, cell pellets were lysed with sodium deoxycholate, and AAV vectors were purified from the cell lysate by ultracentrifugation through an iodixanol density gradient, then concentrated and dialyzed against phosphate-buffered saline (PBS), as previously described ( Cao et al.

Using this model, the authors examined the appearance and progression of tau pathology in diverse areas of the brain in animals of different ages. The results show that tau pathology starts in neurons of the EC expressing the human transgene and over time progresses to cells without detectable human tau expression, first in the vicinity of the

EC and later in more distant regions located downstream in the synaptic circuit, such find more as the dentate gyrus, hippocampus, and cingulate cortex. Human tau protein appears to spread to these brain regions and to interact with and induce aggregation of endogenous mouse tau. The progressive accumulation of tau aggregates leads to synaptic degeneration and later to axonal damage and neuronal death. The exquisite regional specificity of the human transgene expression combined with the use of sophisticated techniques to analyze the brain of these animals enabled the authors to obtain a number of important conclusions, namely: (1) tau aggregates can transfer to neighboring cells and to synaptically connected neurons in distant parts of the brain, all of which do not express detectable levels

of the human protein; (2) misfolded human mutant tau recruits endogenous mouse tau into the aggregates, leading to learn more its progressive intraneuronal accumulation; (3) spreading of tau pathology induces a slow synaptic destruction, followed by axonal and later somatic degeneration of neurons. These are important findings in order to understand the progression of tau pathology and associated damage in AD, and they fit well with recent observations indicating that tau misfolding and aggregation can spread from cell to cell in a prion-like manner (Clavaguera et al., 2009, Frost et al., 2009, Guo and Lee, 2011 and Nonaka et al., 2010). However, a potential weakness of the current study is Ketanserin that, despite

all the diverse techniques used to evaluate human tau expression, the authors cannot completely rule out a low expression (below the level of detection of the methods employed) of the transgene in other brain areas. Indeed, some leakiness of expression has been reported previously for similar mouse models (Santacruz et al., 2005). In this scenario, low widespread expression of human P301L tau, and not spreading of aggregates from one site to another, may have seeded aggregation of endogenous mouse tau and triggered neurodegeneration. Although the authors provide convincing evidence that expression beyond the targeted areas must be very low (or nonexistent), it is also noticeable that because of the high efficiency of the seeding process, these minute quantities may be enough to induce tau aggregation.

A sniff cue (red cross-hair) was then displayed for 667 ms, and recurred with a stimulus-onset asynchrony of 2 s to prompt additional sniffs, as necessary. On the open-sniff trials, subjects made a binary choice with the left or right keyboard arrow once they had accumulated sufficient evidence that clove or lemon was dominating the mixture. Subjects were instructed to emphasize accuracy, ensuring that a decision would be made only when sufficient evidence had been accumulated to the

criterion threshold. This was the primary instruction Afatinib chemical structure given to the subjects. They were incidentally reminded that upon reaching their decision, they should respond by button press as quickly as possible, so that recorded decision times closely reflected the time that they reached their decision. At the end of each trial, subjects also made a perceptual rating on a visual analog scale ranging from pure clove to pure lemon, by moving a cursor from the midpoint of this continuum (representing

equal proportions of the two odors). For the fixed-sniff trials, this estimate yielded binary choice measures according to which side of the midpoint the rating fell on. The next odor was presented 18 s after the end of the previous odor presentation, to minimize olfactory habituation. Binary decisions, analog ratings, and odor presentation times were recorded for each trial. Olfactory and visual stimuli presentations were controlled using Cogent2000 (http://www.vislab.ucl.ac.uk/cogent.php). This was the same as Experiment 1, except that all trials were of the open-sniff www.selleckchem.com/products/MLN-2238.html type. Because this experiment the took place in an MRI scanner, subjects responded using one of two button boxes held in either hand, one representing clove, the other lemon (hand side counter-balanced across runs). These buttons were also used to make the perceptual rating along a visual analog scale. Subjects were not told the outcomes of their decision, to prevent cognitive feedback or reward processing from confounding the neuroimaging findings. Sniffs were visually cued, as before, but were back-projected from a computer monitor

onto a tilted mirror that was affixed to the MRI headbox in front of the subject’s eyes. The letters “L” and “C” (lemon and clove) were presented on opposite sides of the screen to indicate which side represented which odor, and this was counterbalanced across subjects and sessions. Sniff rate was again set at two seconds in order to time-lock this to the data-acquisition rate of the MRI scanner (2,000 ms; see below). Subjects completed two runs of 36 trials on 2 consecutive days (four runs total) to minimize subject fatigue and odor habituation. Each of the nine mixtures was presented eight times each day (144 trials in total over 2 days), and trials were arranged in pseudorandom order such that every mixture preceded every other mixture one time to minimize effects of mixture sequence.

Intake of acetaminophen like drugs and certain chemicals may also lead to hepatocellular carcinoma. N-nitrosodiethylamine (NDEA) is a potent carcinogenic dialkyl nitrosoamine present in tobacco smoke, water, cheddar cheese, cured and fried meals and in a number of alcoholic beverages. It is a hepatocarcinogen producing reproducible HCC after repeated administration. 1 The formation of reactive

oxygen species (ROS) during the metabolism of NDEA may be one of the key factors in the etiology of cancer. 2 HCC is associated with over expression of vascular endothelial growth factor (VEGF) which are produced by hepatocytes in the periportal area of liver tissue. 3 In addition to the animal experimental models of cancer, human cancer cell lines have been widely used to study the antiproliferative effect. AZD2281 datasheet Numerous components of plants, collectively termed “phytochemicals” have been reported to possess substantial chemopreventive properties. Development of nontoxic and biologically safe anticarcinogenic agent has been highlighted as a promising way to treat carcinogenesis.4 Several herbal drugs like Acacia nilotica, Achyranthes aspera, Scutia myrtina, etc have been evaluated for its potential as liver protectant against NDEA

induced hepatotoxicity in rats. 1, 5 and 6 Woodfordia fruticosa (Lythraceae) is a traditional medicinal plant and its dried flowers are used as tonic in Libraries disorders http://www.selleckchem.com/products/Paclitaxel(Taxol).html of mucous membrane, hemorrhoids and in derangement of liver. 7 Phenolics, particularly hydrolyzable tannins and flavonoids were identified as major components of W. fruticosa flowers. In view of these the present work was undertaken to evaluate the protective effect of W. fruticosa against NDEA induced hepatocellular carcinoma in experimental rats and in human hepatoma PLC/PRF/5 cell lines. NDEA, Silymarin, anti-mouse IgG horseradish peroxidase,

streptavidin horseradish peroxidase conjugate, diaminobenzidine, Fetal bovine serum (FBS) and N-2-hydroxyethylpiperazine-N-2-ethane-sulphonic STK38 acid (HEPES) were purchased from Sigma Chemical Co., St. Louis, MO, USA. VEGF antibody from Santa Cruz Biotechnology, Santa Cruz, CA, USA. Alpha feto-protein (AFP) assay kit was purchased from Creative diagnostics, USA. Assay kits for serum alkaline phosphatase (ALP), lactate dehydrogenase (LDH) and bilirubin were purchased from Agappe Diagnostics, India. 5-flourouracil (5-FU) was purchased from Biochem Pharmaceutical Industries, Mumbai, India. RPMI Medium and antibiotic-antimycotic were purchased from Gibco, Grand Island, N.Y, USA. Cell Proliferation Assay kit [3-(4,5-dimethylthiazol-2-yl)-2,5diphenyltetrazoliumbromide (MTT)] was purchased from HiMedia, India. Dimethyl sulfoxide (DMSO) was obtained from Merck, Mumbai, India. All other chemicals were of analytical grade.