Coronal brain sections (40 μm thick) were prepared from injected BAY 73-4506 clinical trial mice at 14 dpi and processed for immunostaining as previously described (Duan et al., 2007 and Ge et al., 2006). For rescue experiments

(Figure 2), anti-GFP antibodies (rabbit, 1:1000, Abcam) were used to enhance the signaling for all conditions in parallel. The sections were incubated for 30 min in 4′,6′-diaminodino-2-phenylindole (DAPI, 1:5000) before washing and mounting. Images were acquired on a META multiphoton confocal system (Zeiss LSM 510) using a multi-track configuration. At least two animals for each condition were analyzed. Statistical comparison of data was performed by ANOVA. For analysis of cell morphology, Z-series stacks of confocal images were taken and a single confocal image slice with the largest

soma area for individual GFP+ neurons was used Screening Library mw for quantification using NIH ImageJ program. For analysis of neuronal positioning, single section confocal images of GFP+ neurons with DAPI staining were used to determine the cell localization within four areas as defined in Figure 1B. For analysis of dendritic development, three-dimensional (3D) reconstruction of entire dendritic processes of each GFP+ neuron was made from Z-series stacks of confocal images. The 2D projection images were traced with NIH ImageJ. All GFP+ dentate granule cells with largely intact dendritic trees were analyzed for total dendritic length as previously described (Duan et al., 2007 and Ge et al., 2006). The measurements did not include corrections for inclinations of dendritic process and therefore represented projected lengths. Sholl analysis for dendritic complexity was carried out by counting the number

of dendrites that crossed a series of concentric circles at 10 μm intervals from the cell soma as previously described (Duan et al., 2007). For analysis of axonal development, cross-sections (50 μm thick) taken perpendicular to the long axis (septal-temporal) were prepared from dissected hippocampi of fixed mouse brain at 14 dpi and processed for immunostaining using anti-GFP antibodies (goat, 1:500, Rockland) and DAPI (1:5000). 3D reconstructions of entire images were obtained using stitch and maximum intensity projection Terminal deoxynucleotidyl transferase functions in the Zen software (Zeiss). For the ZHH sample, the same study group as previously analyzed for DISC1 and NDEL1 interaction were used ( Burdick et al., 2008). Briefly, the cohort included 279 Caucasian patients with schizophrenia or schizoaffective disorder (SZ; 35.5% female) with a mean age of 38.6 ± 10.8 years and an estimated IQ of 96.6 ± 8.5 (based on WRAT-3 Reading). All subjects provided written informed consent to an Institutional Review Board of the North Shore-Long Island Jewish Health System (NSLIJHS)-approved protocol. Clinical characteristics that were collected included duration of illness (17.0 ± 10.8 years) and global assessment of function (GAF; 38.7 ± 15.6).

This is accomplished by nitrosylation (and therefore inactivation) of the GPCR kinases that target the receptors for internalization (Kokkola et al., 2005 and Whalen

et al., 2007). Although the production of eCBs and NO is similarly triggered by a rise in intracellular Ca2+ in the postsynaptic cell, these retrograde signals have opposing actions on GABA release (reviewed in Feil and Kleppisch, 2008 and Wilson and Nicoll, 2002). In this study, we demonstrate that a loss in CB1R signaling is a necessary prerequisite for Sunitinib solubility dmso NO-mediated LTPGABA following acute food deprivation. This finding, that abolition of CB1R signaling is associated with a drive to eat, appears to be at odds with the overwhelming support for an orexigenic role of the eCB system. For example, following acute food deprivation, most evidence points to an increase in hypothalamic eCB levels (Di Marzo et al., 2001 and Kirkham DAPT et al., 2002), and CB1−/− mice exhibit hypophagia compared with their wild-type littermates (Bellocchio et al.,

2010). Recent work, however, provides clear evidence that activation of CB1Rs on GABA terminals is associated with a reduction in feeding. In the ventral striatum, CB1R-mediated inhibition of GABAergic transmission is associated with a hypophagic action of eCBs (Bellocchio et al., 2010). Thus, it is possible that a loss of CB1Rs at GABAergic terminals in the DMH in response to food deprivation is akin to removing the brake from the brain’s feeding circuitry. Our findings provide a demonstration of state-dependent plasticity in the DMH, a hypothalamic nucleus that plays a vital role in integrating information

related to caloric status and stress. Evidence suggests that DMH neurons send glutamatergic projections to the PVN (Thompson et al., 1996), an autonomic nucleus that plays a critical role in regulating food intake (Cowley et al., 1999). In agreement with this, our unpublished observations indicate that the majority of DMH neurons from which we record demonstrate antidromic activation from the PVN. An increase in GABAergic drive in the DMH would therefore 3-mercaptopyruvate sulfurtransferase suppress glutamatergic signaling to PVN neurons that suppress food intake. This signaling would ultimately result in an enhanced drive to eat in the face of minimal food availability. Although stress-induced increases in CORT down-regulate CB1Rs and therefore promote the increase in GABA drive, signaling at the genomic glucocorticoid receptor has no effect on food intake or body weight (unpublished observations). Overall, these data demonstrate a potentially new form of state-dependent plasticity in a feeding circuit that may ensure satiety-sensing neurons in DMH neurons are not recruited when food is not available. All experiments were performed according to protocols approved by the University of Calgary Animal Care and Use Committee in accordance with guidelines established by the Canadian Council on Animal Care.

Thus, O. armillata appears to be protected from eosinophil degranulation, but the mechanism involved for this putatively motile species may differ from that observed in nodule-forming Onchocerca spp. None of the authors declares any conflict of interest. This study was funded by a Wellcome Trust Vacation Scholarship, the British Veterinary Association (BVA) Harry Steele-Bodger Memorial fund, a British Cattle Veterinary Association (BCVA) Student Clinical Research Grant, an Intervet Student Vacation Bursary, a Pfizer Vacation Study Grant

and the University of Cambridge Veterinary School Jowett Fund. The study sponsors did not have any role in the study design; in the collection, analysis and interpretation BVD-523 cost of data; in the writing of the manuscript; or in the decision to submit the manuscript for publication. “
“Vaccines are an important tool in livestock production, not only as a means of maintaining VE-822 mw health and freedom from clinical diseases, but also in some cases as a means of preventing zoonotic disease and thus enhancing food safety and public health. Bovine respiratory

disease (BRD) in calves and young cattle is a significant source of morbidity and mortality, and is a major contributing factor for economic losses in cattle industry (Snowder et al., 2006). BRD is a multifactorial disease (Babiuk et al., 1988); the most common bacterial pathogens associated with it are Mannheimia haemolytica, Pasteurella multocida, Mycoplasma bovis as well as various viral pathogens; bovine respiratory syncytial virus (BRSV), parainfluenza virus type 3 (PI-3), bovine viral diarrhoea virus (BVDV) and bovine herpes virus type 1 (BHV-1). Even though there is limited published information that definitively establishes the efficacy of respiratory vaccines in the field in reducing the burden of bovine respiratory disease ( Perino and Hunsaker, 1997), vaccinations against one or more of the respiratory

pathogens are commonly used. Helminth infections can influence the immune response to unrelated antigens (Kullberg et al., 1992). Furthermore, they have been shown to decrease the response to vaccinations in various host species (Elias et al., 2001 and Urban et al., 2007). Fasciola hepatica, a helminth parasite, causes fasciolosis ALOX15 in cattle and sheep and fasciolosis is also a zoonosis. It is a common parasite, especially in the temperate climate of the UK and Ireland, where the prevalence in cattle is as high as 84% ( McCann et al., 2010). F. hepatica has been proven to have immunoregulatory effects in mice ( Brady et al., 1999). In cattle, it can alter the response to immune-mediated diagnostic tests ( Flynn et al., 2007) but its effect on vaccine responsiveness in this species has not yet been studied. The aim of this study was to establish whether a concurrent F.

Overexpression of CAMKK2 had a similar negative effect on spine density, presumably by increasing calcium sensitization and AMPK activity. The CAMKK2-AMPK pathway appears critical with regard to AD pathology since its blockade mitigates the synaptotoxic effects of Aβ oligomers in vitro and blocks the dendritic spine loss observed in the APPSWE,IND mouse model in vivo. AMPK activity is increased in the hippocampus

of the J20 transgenic mouse model as early as 4 months of age, a time when Aβ oligomer levels are high and signs Cyclopamine concentration of hippocampal network dysfunction already detectable (Palop et al., 2007). Similarly, AMPK activity is increased in the brain of other AD mouse models such as the double APP/PS2 or APPsw/PS1 dE9 mutants at 6 months (Lopez-Lopez et al., 2007; Son et al., 2012), supporting a link between Aβ oligomers and AMPK activation. In agreement with these results, we found that 1 μM Aβ42 oligomer exposure for 24 hr significantly increased AMPK activity in mature cortical cells, confirming previous studies by Thornton et al. (2011). Whether Aβ42 oligomers can activate other members of the AMPK-like family is still unclear, although recent studies report that acute treatment of Aβ42 oligomers does

not activate BRSK2 or MARK3 in primary hippocampal neurons (Thornton et al., 2011). Many kinases can act as direct upstream activators of AMPK, including LKB1 (Hawley et al., 2003; Shaw et al., 2004), CAMKK2, to a lesser extent CAMKK1 (Anderson et al., 2008; Green et al., 2011; Hawley et al., 2005; Hurley et al., 2005; Woods et al., 2005), and TAK1 (Momcilovic

Bleomycin order et al., 2006). We show that Aβ42 oligomer-induced activation of AMPK depends on CAMKK2 in mature synaptically active cortical cultures. Importantly, AMPK is the only member of the AMPK-like family known to be regulated by CAMKK2, whereas other related members of the family are presumably not (Bright et al., 2008; Fogarty et al., 2010). Thus, AMPK may represent the main member of much this family that responds to increased intracellular calcium mediated by NMDAR activation and/or membrane depolarization. Aβ42 oligomer-induced activation of AMPK through CAMKK2 supports the hypothesis that Aβ oligomers may disrupt calcium homeostasis (Demuro et al., 2005; Mattson et al., 1992). Preferential targets of Aβ42 oligomers are dendritic spines (Lacor et al., 2004; Lacor et al., 2007), where they interfere with NMDAR signaling to trigger rise in cytoplasmic calcium (De Felice et al., 2007). Our results provide a mechanism whereby increased neuronal excitation activates the CAMKK2-AMPK pathway leading to Tau phosphorylation on S262 and compromises spine stability. In line with this hypothesis, (1) acute exposure of neuronal cultures to Aβ oligomers leads to local calcium level increase, hyperphosphorylation, and mislocalization of Tau into dendritic spines, which was associated with spine collapse (De Felice et al., 2008; Zempel et al.

Only one study (Galindo-Barboza et al., 2011) has specifically examined the persistence of efficacy of COWP based on worm counts in sheep.

Recent data from goats managed under communal farming conditions suggest that egg counts are reduced two weeks, but not six weeks, after treatment with COWP (Spickett et al., 2012). However, no worm count data are available on the duration of efficacy of COWP in groups of goats subjected to similar levels of parasite exposure, nutrition and management. The present study therefore sought to examine the effect of COWP treatment in goats treated and removed from infective pasture at three different stages, namely at 7, 28 and 56 days post treatment. The use of animals for this experiment met the requirements Thiazovivin in vivo of the Onderstepoort Veterinary Institute Animal Ethics Committee. A 0.67 ha pasture of star grass (Cynodon incompletus Nees) at Onderstepoort Veterinary Institute, Pretoria was utilized for the study in 2006–2007. In the spring of 2006, six months prior to the start of the actual experiment, the grass

was cut and fertilized. The pasture was irrigated through the spring and summer until the conclusion of the experiment in the following autumn if less than 25 mm rain fell during the previous week. Rainfall data were collected at Onderstepoort Veterinary Institute while temperature data were obtained from the IDO inhibitor South African Weather Service for central Pretoria, which is approximately 16 km south of the Institute. Since the pasture had not been used for animal grazing for several years prior to the experiment, it was seeded with H. contortus larvae by grazing infected sheep on it. Initially, twenty indigenous check sheep were purchased from a commercial vendor, transported to Onderstepoort Veterinary Institute and maintained in concrete pens which were swept clean daily to preclude accidental nematode infection. The

animals were fed a commercial pelleted feed and lucerne (Medicago sativa) hay and the animals had free access to water. The sheep were dewormed with 10 mg/kg fenbendazole (Panacur BS®, Intervet South Africa) and 7.5 mg/kg levamisole (Tramisol®, Coopers, Afrivet Business Management, South Africa) daily for 5 days, followed by 0.3–0.5 mg/kg ivermectin (Ivomec Injection®, Merial South Africa) administered 6 days and 13 days after the combination treatment with fenbendazole and levamisole. Thirty-three days later, the animals were infected with 5000 third-stage larvae of a susceptible strain of H. contortus given as 1000 larvae per day for five days, as low-level, trickle dosing has been shown to be the optimal method for achieving establishment of parasites ( Barger et al., 1985 and Dobson et al., 1990).

5% ± 7.0% reduction in amplitudes, n = 7). In the miniSOG-VAMP2 expressing cells, a decrease of 29.4% ± 3.3% (n = 4, p = 0.006) in EPSC amplitude was observed after light illumination (Figures 1B and 1C). In the SYP1-miniSOG expressing cells, the reduction of EPSC amplitude was significantly greater at 82.6% ± 8.5% (n = 6, p = 0.0002). In

two of the cells that exhibited the reduction of EPSC with SYP1-miniSOG, a stable recording was maintained for 45 min to 1 hr and no recovery in EPSC amplitude was observed. The reduction of EPSC amplitudes was associated with insignificant changes of electrophysiological properties such as membrane BAY 73-4506 resistance (105.6 ± 35.6 MΩ to 73.6 ± 23.2 MΩ, n = 6, p = 0.20) or capacitance (62.5 ± 13.9 pF to 56.9 ± 6.7 pF, n = 6, p = 0.55). The reduction of EPSC amplitude did not alter the ability of the cell to fire action potentials in response to depolarizing current injections (Figure 1D; n = 5). Active and continual synaptic release during illumination was not essential for the inhibition of synaptic release, as the inhibition was still observed in cells where the self-stimulation was discontinued during light illumination (63.7% and 45.7% inhibition in two cells tested with SYP1-miniSOG). In order to examine whether we can selectively inhibit specific synapses with high spatial resolution, we performed an imaging-based assay with the membrane dye FM4-64 in combination with SYP1-miniSOG. In this assay, SYP1-miniSOG fused to eGFP

or Citrine was expressed in cultured cortical neurons. A DNA Damage inhibitor quadrant of the field of view (159 μm × 159 μm) was scanned with 488 nm laser to conduct CALI of the presynaptic terminals. Vesicular release was induced with 40 mM KCl solution containing 10 μM FM4-64. The high potassium solution and dye solution was subsequently washed out to repolarize the membrane and remove membrane bound FM4-64 (Cousin, 2008). We then

measured and averaged the FM4-64 fluorescence of the puncta positive for both FM4-64 and SYP1-miniSOG-eGFP/Citrine inside and outside the CALI region (Figure 1E). The SYP1-miniSOG-eGFP/Citrine positive puncta within the CALI region had significant less FM4-64 dye uptake compared to the puncta outside the CALI region (5911.2 ± 687.5 and 8118.3 ± 763.2 arbitrary units, n = 78 and n = 95, respectively; p = 0.037). However, this measurement is likely an underestimate of the true level Astemizole of synaptic inhibition, as we only quantified puncta that are positive for both eGFP/Citrine and FM4-64 and omitted eGFP/Citrine positive puncta that did not have FM4-64 fluorescence. This selection was necessary as not all fluorescent protein-positive puncta were functional presynaptic boutons capable of dye uptake even in the absence of light illumination (see Figure S1 available online). No significant difference was detected between the puncta inside and outside the CALI region in SYP1-eGFP positive puncta (6562.6 ± 466.9 and 7551.1 ± 560.6 arbitrary units, n = 114 and n = 157, respectively; p = 0.

Interestingly, the degree of correlation was dramatically reduced when the burst firing was filtered out from analysis, indicating that the Selleck ABT-263 synchrony was a reflection of the abnormal burst firing. The pooled data from the recordings

in five intact and eight hemi-PD rats confirmed this observation (Figure 4B). Power spectrum analysis of the local field potential (LFP) revealed the appearance of oscillatory activity at the beta band (∼20–30 Hz) after dopamine depletion (Figures 4C and 4D). STN-DBS at 125 Hz was highly effective in removing the abnormal synchrony among the neurons and the beta oscillations (Figures 4B and 4D). The strength of coupling between spike times and LFP at any given frequency, known as spike-field coherence, was also investigated. 6-OHDA lesion caused the coherence value to increase specifically at the beta band (20–30 Hz; Figures 5A and 5B). Interestingly, the delivery of 125 Hz STN-DBS eliminated this beta band spike-field coherence. In addition, the phase synchronization (or coherence phase) between spikes and LFP at the beta band was derived and presented as polar histogram (Figure 5C). It was clear that the 6-OHDA GDC-0941 purchase lesion induced a specific phase-locking phenomenon between spikes

and LFP in beta band (91/115 pairs, eight rats), whereas the pairs from the intact and unlesioned side showed randomly distributed phases, ranging much from 0° to 360° (intact: 88 pairs from five rats; unlesioned: 98 pairs from eight rats). During the delivery of 125 Hz STN-DBS, the specific phase-locking bias present in the 6-OHDA-lesioned condition was not notable. To elucidate the possible mechanism underlying antidromic spikes-mediated beneficial effect, we next asked the crucial question of whether antidromic spikes can directly modulate the firing properties of the CxFn. Since the DBS was delivered at a high frequency of 125 Hz, i.e., with an interstimulus interval of 8 ms, we analyzed the impact

of STN-DBS by selecting and then aligning all those 8 ms time segments that contained antidromic spikes. A typical example is shown in Figure 6A (left panel). These time-aligned segments of neuronal activities revealed that the probability of firing of the CxFn was influenced by the 125 Hz HFS: a complete cessation of firing immediately following the antidromic spike that lasted for about 1 ms followed by an elevated firing probability in the subsequent 2 ms interval. The early depression of firing probably represents the refractory period of the antidromic spikes, while the delayed increase in firing probability could reflect a change in the intrinsic excitability of the neurons or functional connectivity within the local circuit as a consequence of antidromic spikes. In contrast, for those 8 ms segments that did not contain antidromic spikes, there was no change in the firing probability (Figure 6A, right panel).

In addition to influenza, pharmacists have also become significant providers of Tdap vaccinations [29]. Pharmacists are currently authorized to administer Tdap vaccinations under a protocol or with a patient specific prescription in 43 states and the District of Columbia [30]. On the Northwestern Memorial Hospital (NMH) campus, Prentice Women’s Hospital (PWH) delivers 10,000–12,000 babies each year. PWH DAPT research buy has implemented and achieved success with a program to vaccinate postpartum women; they reported 78.87% of postpartum patients received the Tdap vaccination between June 2008 and November 2009 [31]. The objective

of this study is to investigate the rate of Tdap vaccination among close contacts of neonates in a women’s hospital pharmacy and to assess the impact of a coordinated pharmacy

and hospital Tdap vaccination program. Walgreens operates a retail pharmacy on the Northwestern Memorial Hospital (NMH) campus. The pharmacists at this location are certified immunizers and maintain an ample supply of Tdap vaccine. While the Prentice Women’s Hospital (PWH) has achieved a high vaccination rate of postpartum patients, the number of close contacts receiving the Tdap vaccination at the retail pharmacy has been minimal. On occasion, some fathers and close contacts presented Kinase Inhibitor Library to the pharmacy to request the vaccine, which was administered under a standing order protocol. On December 9, 2010, Walgreens and PWH implemented a program to increase Tdap vaccination uptake among close contacts of neonates through educating this population on the importance of receiving the vaccine and referring them to the pharmacy for vaccination. Prior to this initiative, there was no formal education or referral for close contacts

of neonates. Educational materials regarding the risks of pertussis, importance of the Tdap vaccination, and promotion of the hospital vaccination clinic were added to the existing admission packet given to delivering families. Also included in the admission packet were a vaccine administration record (VAR) and vaccine information sheet (VIS). These materials included the time and location of pharmacist daily vaccination clinics. For up DNA ligase to two hours each weekday, an on-site pharmacist held a pertussis vaccination clinic at PWH. The entire staff of the delivery unit was educated on the program and was responsible for its promotion. Pharmacists and staff were available to respond to any questions from patients. This cross-sectional study analyzed all Tdap vaccinations administered at the Walgreens pharmacy located on the Prentice Women’s Hospital campus (intervention pharmacy with in-hospital vaccination) between December 2008 and November 2012. The pre-study period was defined as 24 months prior to initiation of the program, with Tdap vaccination claims administered from December 2008 through November 2010.

, 2011). Second, they labeled a small number of sister cells with retroviruses in a late phase of embryonic development (embryonic days [E] 15–E17), while we labeled the entire progeny from a single progenitor starting about E12 (Magavi et al., 2012). The sister pairs analyzed in their study were, on average, more closely related in lineage than two randomly selected cells in a clone labeled in our

mice, which could result in different degrees of shared orientation selectivity. Third, they examined only vertically aligned pairs, while we examined all possible pairs in a clone. It is not clear yet whether vertical alignment affects functional similarity. Fourth, they reported that retrovirally infected cells were much more responsive (34 responsive pairs in 52 pairs; 65% of pairs) than the entire selleck chemical population (38% of neurons, or 14% of pairs assuming independent selection), suggesting that retrovirus infection might have affected the responsiveness of the infected neurons. Our findings may explain the salt-and-pepper functional architecture in rodent visual cortex. In mice, neurons derived from the same progenitors tend to share orientation preference, and neurons derived from different MAPK inhibitor progenitors are spatially intermingled. This distribution

of clonally related neurons may work as the scaffold to generate the salt-and-pepper architecture observed in rodents. If so, could lineage also account for the architecture of the homogeneous through functional columns (Hubel and Wiesel, 1962 and Hubel and Wiesel, 1968) observed in higher mammals, such as carnivores and primates? The distribution of clonally related cells seems less laterally dispersed and more radially aligned in the monkey cortex (Kornack and Rakic, 1995; but cf. more laterally dispersed

clones in the ferret cortex, Reid et al., 1997), but the complete picture of the progeny of single progenitors has not yet been described. In higher mammals, a large expansion of the subventricular zone has been reported, with each progenitor giving rise to a very large number of neurons through intermediate progenitors (Kriegstein et al., 2006 and Lui et al., 2011). In this scenario, individual cortical stem cells in higher mammals may produce a large cohort of neurons that may comprise an entire functional column with little intermingling of neurons derived from other clones. Alternatively, in higher mammals, each single functional column may be derived from multiple clones (Rakic, 1988 and Rakic, 1995), and some mechanisms may group neighboring neurons (Yuste et al., 1992) derived from multiple clones to give rise to their homogeneous functional columns. Procedures are described in detail in Supplemental Experimental Procedures. Z/EG (Novak et al., 2000) and Ai9 (Madisen et al., 2010) mice were obtained from the Jackson Laboratory. TFC.09 mice were generated by enhancer trapping, in which the minimal promoter of mouse Thy-1.2 gene regulates Cre recombinase expression ( Magavi et al., 2012).

1 It is found in wooded areas of Senegal, southern part of Nigeria, Central and Eastern Africa. 2 It is used for the treatment of backache, diabetes and as an anti-scorbutic. The leaves of the plant boiled in its own sap are used for the treatment of gastrointestinal sores. 1 Its sap is used for toothache and cough. 3 It is used in the treatment of jaundice and haemorrhoids among the Baka Pygmies of Cameroon and also used in the traditional

treatment of inflammatory, skin infection and ulcer. 4 and 5 The presence of alkaloids, tannins, saponins, phlobatannins, terpenoids and flavonoids in the leaves of T. potatoria has been reported. 6T. potatoria root has also been found to contain phytochemicals such as tannins, flavonoids, phlobatannins and cardiac glycosides. 7 Betulinic acid, 3β-hydroxy-lup-20(29)-en-28-oic acid, a C-28 carboxylic acid derivative of the ubiquitous triterpene BMS-777607 research buy betulin, is a member of the class of the lupane-type pentacyclic triterpenes. Figure options Download full-size image Download as PowerPoint slide It was isolated at the beginning of the 20th century and originally called gratiolone.8 However unlike betulin, the oxidized derivative

ON-01910 mw betulinic acid possesses a number of intriguing pharmacological effects including: anti-inflammatory, anticancer and anti-HIV.5, 9 and 10 T. potatoria root was collected from Ilesa, Osun state, Nigeria and authenticated by Mr. G. Ibhanesebhor, plant taxonomist, Herbarium, Obafemi Awolowo University, Ile-Ife, Nigeria. Voucher specimen (IFE Herbarium 16419) was deposited in the herbarium. The plant material those was air-dried, pulverised

and extracted by soaking 1.2 kg sample in aspirator bottles containing distilled methanol at room temperature (25 °C) for 48 h. The extract was filtered and solvent was completely removed by vacuum evaporator at 50 °C to give viscous mass (18.55 g, 1.5% yield), which was stored inside a dessicator for further usage. Phytochemical screenings of MeTp were performed using standard procedures.11, 12 and 13 0.5 g of the extract was boiled with 10 ml of sulphuric acid (H2SO4) and filtered hot. The filtrate was shaken with 5 ml of chloroform. The chloroform layer was pipetted into another test tube and 1 ml of dilute ammonia solution was added. The presence of pink colour in the aqueous layer indicated the presence of anthraquinones. 5 ml dilute ammonia was added to a portion of an aqueous filtrate of the extract. Concentrated sulphuric acid (1 ml) was added. A yellow colouration that disappears on standing indicates the presence of flavonoids. About 0.5 g of the extract was boiled in 10 ml of water in a test tube and then filtered. A few drops of 0.1% ferric chloride was added and observed for brownish green or a blue-black colouration. To 0.5 g of extract was added 5 ml of distilled water in a test tube. The solution was shaken vigorously and observed for a stable persistent froth.