a edição); AJCC –estádio IV. No 10.° dia pós-operatório, o doente desenvolveu quadro de dispneia progressiva e febre, associado a hipoxemia e aumento

dos parâmetros inflamatórios (leucócitos 17,5 G/L; PCR 20,8 mg/dL). A radiografia do tórax e a TC com contrate endovenoso identificaram a presença de tromboembolia pulmonar bilateral, uma pneumonia do lobo inferior esquerdo e a existência de 2 coleções intra-abdominais, uma posterior à cauda do pâncreas (com 6 cm de maior diâmetro) e outra retropancreática e estendendo-se até ao bordo hepático, de configuração alongada (com 2 x 13 cm). Foi mantida a drenagem abdominal externa por drenos multicapilar, iniciando-se antibioterapia ev de largo espectro (piperacilina + tazobactam 4.500 mg 3 id e vancomicina 1.000 mg 2 id) e anticoagulação em dose terapêutica (enoxaparina ZD1839 60 mg sc 2 id). A introdução destas medidas levou a uma melhoria clínica e laboratorial, mantendo-se, contudo, as 2 coleções intra-abdominais com características sobreponíveis à avaliação imagiológica inicial. O doente teve alta (ao 39.° dia pós-cirúrgico), sob anticoagulação oral e com revisão Galunisertib molecular weight imagiológica programada. Uma semana após a alta, o doente foi readmitido por um quadro de tosse, dispneia, febre e dor abdominal. Analiticamente apresentava novamente leucócitos e PCR aumentados (18,4 G/L;

21,7 mg/dL, respetivamente). Na ecografia abdominal era evidente a manutenção de coleções intra-abdominais, de localização retropancreática e subdiafragmática, agora com algumas bolhas gasosas, sugerindo a presença de fístula intra-abdominal. Apesar da instituição de medidas agressivas de suporte, terapêutica antimicrobiana e antifúngica de largo espectro, o doente apresentou uma rápida evolução desfavorável, com

sépsis grave e falência multiorgânica (insuficiência respiratória parcial e falência circulatória). A TC toraco-abdominal demonstrou a presença de solução de continuidade transdiafragmática (fig. 1a) entre as coleções abdominais previamente existentes e um abcesso da base pulmonar esquerda (fig. 1b). A realização do trânsito esófago-jejunal contrastado revelou extravasamento de produto de contraste para as coleções abdominais e destas para a árvore brônquica Metalloexopeptidase esquerda (fig. 1c). A avaliação endoscópica permitiu identificar uma anastomose esófago-jejunal íntegra, mas no fundo da ansa cega do Y-de-Roux constatou-se a existência de uma deiscência com cerca de 1 cm, com bordos inflamados, espessados e de aspeto fibrosado, prolongando-se por orifício fistuloso (fig. 2a). A realização de laparotomia exploradora foi afastada pelo elevado risco cirúrgico. A ausência de alternativa cirúrgica e o agravamento progressivo do quadro clínico, conduziu à tentativa, até então não considerada, de resolução do quadro através de métodos endoscópicos.

This group found that the expression of these receptors is restricted to tumorous prostate tissues whereas the B2 appeared more widely expressed in normal and diseased prostate [52]. In brief bradykinin antagonists are under investigation as new antitumoral drugs and the B1 receptor appears

to be a potential target for adjunctive therapy of hormone-refractory prostate cancers. The major advantage of a combination in cancer chemotherapy in a unique agent blocking all features of cancer growth stimulation is the aim of several investigators [50]. In search of more potent and more selective bradykinin antagonists XL184 research buy as potential anticancer agents, the effects of R-954 in mouse and rat models of Ehrlich tumor were evaluated. All experiments were performed with male Balb/C mice (20–25 g) or male Wistar rats (150–200 g) obtained from our own KU-57788 purchase animal facility. Animals were maintained in a room with controlled temperature 22 ± 2 °C for 12 h light/dark cycle, with free access to food and water. Animals were killed in a chamber with saturated CO2 atmosphere to avoid hemorrhage in the peritoneal cavity. Animal care, research and animal sacrifice protocols were in accordance with the principles and guidelines adopted by the Brazilian College of Animal Experimentation (COBEA), were

approved by the Biomedical Science Institute/UFRJ Ethical Committee for Animal Research, and received the protocol number ICBDFBC-015. The bradykinin B1 receptor antagonist R-954 (Ac-Orn-[Oic2, a-Me Phe5, D-b Nal7, Ile8] desArg9 bradykinin) [36] was dissolved in sterile phosphate buffer saline (PBS) and administered subcutaneously at the dose of 2 mg/kg in a final volume of 0.1 ml per animal. Vincristine sulfate (Sigma Chem., St Louis, MO, USA) was used at the optimal

concentration of 0.5 mg/kg for comparison purpose. The control group was given the vehicle (PBS). Mice and rats were given R-954 or vehicle every 24 h after inoculation of Ehrlich ascitic tumor cells until the end of experiment. Ehrlich ascitic tumor (EAT) cells derived from a spontaneous murine mammary adenocarcinoma, were maintained in the ascitic form by sequential passages in Balb/C mice by means of weekly i.p. transplantations of 5 × 105 tumor cells. For the experiments on ascitic pheromone tumor, mice were given an i.p. inoculation of 5 × 105 tumor cells in 0.5 ml and were sacrificed 10 days after. Samples of blood, bone marrow lavage and ascitic fluid were colleted for several measurements as described. For the series of experiments on rat solid tumor, 5 × 105 tumor cells were injected in a volume of 0.1 ml in the footpad of rats and the contralateral paw was administered the vehicle [19]. Every 24 h and until the 7th day, the paw edema was measured by pletismography as described in [16]. Bone marrow cells were obtained by flushing the femoral cavity with 1 ml of PBS. A blood aliquot was collected for cell count.

All patients had negative laboratory, cross sectional imaging and routine endoscopy. EUS was performed with Olympus radial echoendoscope. SOM was performed during ERCP using a triple lumen perfusion catheter

with evaluation of the appropriate sphincter. Sedation was performed under general anesthesia. In the patients presenting with ARP (n=150), 87 (58%) had abnormal studies, including 66 with features suggestive or diagnostic of chronic pancreatitis (Rosemont Criteria) and 6 patients with pancreatic neoplasm (adenocarcinoma, neuroendocrine tumors, lymphoma), pancreatic cysts (9). In patients presenting with PCS (n=207), only 27 (13%) had abnormal studies including 15 with features suggestive of Chronic Pancreatitis, Venetoclax 6 with bile duct stones and 6 with dilated ducts (pancreatic and or biliary). In patients presenting with pain only (n=165), 33 (20%) had abnormal studies including 22 with features suggestive of Chronic Pancreatitis, 1 each with pancreatic neoplasm and bile duct stone, 3 with pancreatic cysts and 5 with dilated ducts. Patients presenting with ARP or obscure abdominal pain with or without prior cholecystectomy may benefit from Endoscopic Ultrasound.

Advanced endoscopic procedures such as ERCP with SOM are often requested in this group of patients. Simultaneous ERCP/SOM and EUS results in establishing a final diagnosis much superior than either 6-phosphogluconolactonase study alone. This results in more expedient completion of medical testing and institution of appropriate management in these challenging selleck inhibitor groups of patients. The group most likely to benefit from both studies are those presenting with ARP. Category Patient (n) EUS EUS Findings NL (%) ABN (%) CP PN BDS DBD/PD PC SMT ARP 150 63 (42%) 87 (58%) 66 6 2 4 9 0 PCS 207 180 (87%) 27 (13%) 15 0 6 6 0 0 Pain 165 132 (80%) 33 (20%) 22 1 1 5 3 1 Total

522 375 (72%) 147 (28%) 103 7 9 15 12 1 CP= Chronic pancreatits; PN= Pancreatc Neoplasm; BDS= Bile Duct Stones DBD/PD= Dialted Biliary Buct/Pancreatic Duct; PC= Pancreatic Cyst; SMT= Submucosal Tumor “
“Currently, there are no quality measures specific to children undergoing gastrointestinal endoscopy. To determine the baseline quality of pediatric colonoscopy reports, key quality indicators must be monitored and analyzed. The consortium of the Pediatric Endoscopy Database System-Clinical Outcomes Research Initiative (PEDS-CORI) use a structured computerized endoscopy report generator, which includes fields for specific quality indicators. We conducted prospective data collection using a standard computerized report generator and central registry (PEDS-CORI) to examine key quality indicators from 14 pediatric centers between Jan 2000 and Dec 2011. Reports were queried to determine if specific quality indicators were recorded.

, 1984; Gutierrez and Ownby, 2003). Conventional antivenoms are prepared by immunizing horses with venom from a single snake species or a mixture of venoms from different species. The aim of immunization is to elicit high levels of antibodies that bind to and neutralize most relevant toxins. Conversely,

immunization also elicits undesirable antibodies directed to non-toxic venom components and irrelevant venom epitopes, AZD9291 purchase according to Harrison et al. (2011) 95% of IgGs comprising current antivenoms are not therapeutic. All the irrelevant proteins contribute to some antivenom therapy side effects. For instance, even though immunoglobulin G(T) is effective in the treatment of envenomed patients, a high incidence (37–87%) of early anaphylatic reactions requiring urgent treatment with adrenalin and antihistamines have been observed (Cardoso et al., 1993). Mixtures containing mono-specific antibodies against a repertoire of epitopes in toxic venoms could help achieve two desirable immunotherapy requirements: the use of smaller amounts of antivenom, and higher specificity. In addition, the development JNJ-26481585 cost of bothropic antivenoms should consider the need to reduce components other than the desired venom-specific IgG or their F(ab′)2 fragments and the use of a mixture of antibodies restricted to the relevant toxic venom components. The aim of our work was to develop in mice monoclonal antibodies against some B. atrox venom components.

Their

neutralizing properties were analyzed using some well known pathological process induced by venom components as indicators. Three specific neutralizing mAbs (thrombin-like 6AD2-G5 clone, PLA2 A85/9-4 clone, and Zn-metalloprotease 59/2-E4 clone) were prepared and tested by their ability to neutralize the main B. atrox venom toxins. These monoclonal antibodies will be used selleck products in the future as raw reagents to prepare hybrid antibodies expressing the mouse LV and HV regions molecular linked into human LC and HC regions. B. atrox venom was kindly provided by the Laboratório de Hepertologia, Instituto Butantan, São Paulo, SP, Brazil, in the lyophilized form. The venom used in this work is a pool of snake venom from Tucuruí, Pará, Brazil. Venom was weighed, diluted in distilled water to a final concentration of 10 mg/ml, and stored at −20 °C. Bothropic antivenom (batch 0512219/B, expiry date April 2009) was provided by Instituto Butantan. Swiss mice weighing between 18 and 22 g were used throughout this study. Male and female adult BALB/c mice were also used. Animals were bred at the Vivarium of Isogenic Mice at the Center for Biosciences and Biotechnology of Universidade Estadual do Norte Fluminense Darcy Ribeiro (UENF). All animals were housed in controlled rooms and received water and food ad libitum until used. When necessary, 250 μg of ketamine chloridrate were used to anesthetize mice. This study was approved by the Experimental Animals Committee of UENF.

Written informed consent was obtained in all patients who participated in this study. An admission blood sample was provided by patients followed by serial samples at 1, 4, 12, and 24 h, then once daily until discharge or death, as allowed by clinical factors. Blood was collected into an EDTA tube which was promptly centrifuged and the plasma was removed and frozen at −23 °C until the time of analysis. Ethics approval for this observational study was obtained

from Sri Lanka (the Universities of Colombo, Peradeniya and Sri Lankan Medical Association) and the grantholder’s universities (Oxfordshire Clinical Research Ethics Committee (UK) and Australian Osimertinib manufacturer National University). The total and free SB203580 (unbound) concentrations of MCPA were quantified in the samples collected above in addition to admission samples collected for a previous study (Roberts et al., 2005). The total MCPA concentration was measured in the above-mentioned plasma samples. 300 μL of plasma was then ultrafiltered using Millipore Centrifree Micropartition Device® (Millipore, Bedford, MA, USA) yielding approximately 100 μg of plasma ultrafiltrate. The concentration of MCPA in the ultrafiltrate is the free (unbound) concentration. The concentrations of MCPA were determined by Queensland Health

Scientific Services (Australia) using a method derived from that of the United States Environmental Protection Agency (EPA, 1980). 100 μg of plasma or ultrafiltrate was hydrolysed in diluted sodium hydroxide and then buffered with acetic acid. The concentration of MCPA was determined by HPLC–MS/MS using an AB/Sciex API4000Q mass spectrometer in the negative ion mode equipped with an electrospray (TurboV) interface. This was coupled to a Shimadzu Prominence HPLC system (Shimadzu Corp., Kyoto, Japan) and a 50 mm × 2 mm C6-phenyl column (Phenomenex, Torrance, CA). The limit of reporting for the LCMSMS method was 1 μg/L for MCPA and the method was linear from at least 1–300 μg/L. Method recovery was confirmed using MPCA concentrations of 2.5 mg/L to around 300 mg/L with an average recovery of 105% and a standard deviation 0.25. Therefore, the limit

of detection (LOD; 3× standard GBA3 deviation) is 0.75 mg/kg and the limit of reporting (LOR; using 6× LOD) is 4.5 mg/kg. The resulting concentrations (mg/kg plasma) were multiplied by 1.0205 which is the specific gravity of plasma at 37 °C (Trudnowski and Rico, 1974) to allow reporting with the unit mg/L. To validate the Centrifree® ultrafiltration device, plasma from a patient with MCPA poisoning was ultrafiltered, analysed for MCPA, re-ultrafiltered and then re-analysed for MCPA. There was no change in the concentration of MCPA between the 2 ultrafiltrates so MCPA does not appear to be adsorbed to the ultrafiltration device. Further, control plasma which did not contain MCPA was ultrafiltered and the filtrate was analysed for protein. It was noted that <0.

In 2003, a project was initiated to assess reactions to 11 major diseases of maize inbred lines that are used in current breeding programs. The objective of the present study was to evaluate the reactions to NCLB, SCLB, CLS, GLS, common rust, and southern rust of a collection of parental inbred lines that are actively used in most maize breeding programs or are widely find more grown cultivars. One hundred and fifty-two inbred lines of maize were collected from the major maize breeding

programs in China and the seeds were increased at the Maize Centre, Chinese Academy of Agricultural Sciences (CAAS), Beijing, China. Based on information of their pedigrees and genetic structures [19], [20] and [21], 129 inbred lines were categorized into heterotic group A or B. Group A contained subgroups PA (group A germplasm derived from modern U.S. hybrids) (30 lines), BSSS (Iowa Stiff Stalk Synthetic population) (25 lines), and LRC (derivatives of Lvda red cob Chinese landrace) (19 lines); and group B consisted of subgroups PB (group B germplasm derived from modern U.S. hybrids) (18 lines), Lan (Lancaster Surecrop) (17 lines), and SPT (derivatives of Tangshan Sipingtou Chinese landrace) (20 lines). Twenty-three lines were not assigned to any subgroup, owing to a lack of pedigree or molecular

genetic information (Table 1). For accurate evaluation of disease reactions under appropriate environments, the screening nursery was located in disease epidemic areas: the NCLB nursery was in Harbin, Heilongjiang province; SCLB and CLS nurseries were in Beijing; GLS and common rust nurseries were in Shenyang, BLZ945 research buy Liaoning province; and the southern rust nursery was in Sanya, Hainan province in

the winter growing season. The first screen for resistance to NCLB, SCLB, CLS, GLS, and common rust was conducted in 2003 and 2004 for 106 and 46 lines, respectively, and was repeated from 2004 to 2005. Reactions to southern rust were evaluated in 2004 and repeated in 2005. Seeds were planted on the farm of the Heilongjiang Academy of Agricultural Sciences (HAAS), Harbin, Heilongjiang province, China. The inbred lines Mo 17 and Huobai were used as resistant and susceptible controls, respectively. Race 1 of E. turcicum was amplified on sorghum (Sorghum bicolor [L.] Moench) grain medium [22] these at 23–25 °C in the dark to promote sporulation. Spores were suspended in distilled water at concentrations of 1 × 105 mL− 1 to 1 × 106 mL− 1 before inoculation. At growth stage V10 [23], inoculation was performed by spraying approximately 10 mL of spore suspension onto the leaf surfaces of each plant. Seeds of each line were grown on the experimental farm of the Institute of Crop Science, CAAS, Beijing, China. Lines Mo 17 and Luo 31 were used as resistant and susceptible controls for assessment of SCLB reactions, and Shen 137 and Huangzaosi were grown as resistant and susceptible controls for evaluation of reactions to CLS.

Fishers average weekly takings after fuel costs, ranged from $US 450 to 3150 for fish ($US 1671±730) and from $US 210 to 1753 for lobster ($US 836±458), highlighting the profitability of fishing in Anguilla. The most recent hurricanes

that severely impacted Anguilla are hurricanes Luis in 1995 and Lenny in 1999. Hurricane Lenny caused significant flooding and damage to land-based infrastructure, but less impact at sea or on the fishing community. Consequently, when recounting impacts suffered from hurricanes, respondents NSC 683864 cost predominantly focused their responses to the effects of hurricane Luis (Table 1). The accuracy of these recollections may be enhanced by both the age of these fishers and that many were fishing during hurricane Luis, in addition to the general significance of hurricane Luis for the whole island. The majority of respondents (75%) lost gear (fish and/or lobster traps) as a consequence of hurricane Luis, with losses per fisher ranging from 13 to 250 (mean±SD, 86±67) traps. The combination of lost gear and the impact of the hurricane on hotels meant that fishers were unable to fish for at least two months (Table 1),

although one fisher stated he did not return to fishing for approximately three years. Respondents stated that the Anguilla government provided some financial assistance to the fishing community by giving each

fisher three traps to re-start fishing, and offering subsidies on wire mesh and buoys to help fishers rebuild traps. In addition to the substantial financial Fasudil in vitro impacts accrued, six respondents stated that the fishing grounds had been altered by the hurricane. Another six respondents mentioned that the fishing grounds had been completely destroyed. All respondents continued to fish after the devastation of this hurricane, even though some took several years to return to fishing. It would appear that, despite the destruction of the Fenbendazole hurricane, fishing remained a viable occupation, and the profitability of fishing in Anguilla will likely have influenced the decision of these fishers to continue fishing. The personal and cultural ties that fishers have with their occupation, their ‘fisher ethic’, may provide an additional explanation for why fishers continued to fish after hurricane Luis. When asked why they fished, 63% (n=15/24) of respondents stated their motive was because of an ingrained cultural or personal desire to fish. By comparison, fewer respondents (33%, n=8/24) mentioned the financial motivation. Examples of respondent response categories and selected quotes illustrating ‘fisher ethic’ are shown in Table 2. The impact of hurricane Luis was manifest in seasonal changes in the fishing practices on Anguilla.

However, it is reasonable to assume that some other mechanisms may be in place in non-proliferating cells in which no telomeric attrition due to the end replication problem is expected to occur, either because these cells are quiescent or differentiated. Surprisingly however,

we and others have shown that telomeres might have a central role in senescence establishment independently from their shortening [ 36•• and 37••]. In these reports, random DNA damage generated by ionizing radiation, genotoxic drugs, or H2O2, leads to DDR Nintedanib cost activation that preferentially persists at telomeres over time. Cells with persistent DDR activation show a senescent phenotype that cannot be prevented by exogenous expression of telomerase, further excluding a contribution of telomere shortening. The mechanism proposed to explain this phenomenon is the suppression of effective DNA repair at telomeres by TRF2, a telomeric DNA binding protein [ 36••]. Inhibition of DNA repair might reflect the evolutionary role of check details telomeres in preventing chromosomal fusions, illegitimate DNA repair events among chromosome ends, in order to maintain the linear structure of chromosomes. TRF2 and the associated RAP1 protein are indeed able to inhibit NHEJ in vitro [ 38, 39 and 40]

and knock out of TRF2 leads to dramatic chromosomal fusions [ 41 and 42], most of which depend on NHEJ [ 43•]. Similarly, TRF2 has been shown to inhibit NHEJ also when a DSB occurs within a telomere, and not only at its end ( Figure 1), revealing that telomeric proteins, rather than telomeric DNA, are responsible for telomere irreparability. Consistent with this model, DDR activation at telomeres is more frequent in mouse and baboon tissues from aged animals, when compared with their young counterparts [ 36•• and 37••]. This observation also suggests that having long telomeres

may have an important drawback, since more telomeric DNA can offer a wider target for random DNA damage that cannot be repaired. Indeed, in different mammalian species, telomere length and lifespan are inversely correlated [ 44]. In addition to its potential role in promoting ageing and age related Exoribonuclease disorders, telomere-initiated senescence, fuelled by oncogenic signals, plays a prominent role in suppressing malignant cancer progression in humans. In cells with functional DDR, oncogene expression usually results in cellular senescence after just a few population doublings [45]. This proliferative arrest is called oncogene-induced senescence (OIS) and, depending on cell type and oncogene expression levels, is caused by activation of a number of diverse pathways [46]. Thus, by preventing cancer onset, in addition to causing impairment of regenerative capacity during ageing, cellular senescence has been considered as an example of antagonistic pleiotropy, although this has recently put to question [47].

The study conforms to the APS Guiding Principles for the Care and Use of Animals in Research. A total of 20 ESDs were performed (10 with each technique). The dissection time was shorter with HK-CB (4.9±3.2 vs. 13.8 ±10.8 min; p=0.002), even if the time to apply the CB device was included (9.0±6.5 vs. 13.8±10.8 min; p=0.008). The dissection speed was faster with HK-CB (0.8±0.4 vs. 0.4±0.3 cm2/min; p=0.014), No differences were observed in the remaining variables. There was one perforation in each group. The CB traction method shortens the duration of the dissection phase of gastric ESD in a live porcine model. This method may facilitate the introduction of ESD

especially in the beginning of the learning curve. Z-VAD-FMK datasheet “
“After non-curative endoscopic submucosal dissection (ESD) for differentiated-type early gastric cancer (EGC), if a positive lateral margin (LM) or piecemeal resection is the only non-curative factor, non-surgical management (close observation or immediate additional endoscopic treatment) can be performed instead of gastrectomy due to the negligible risk

of metastasis. However, the most appropriate management is unknown because of http://www.selleckchem.com/products/fg-4592.html limited research. To examine the long-term outcomes and risk factors for residual/recurrent cancer in non-curative ESD cases of differentiated-type EGC with a positive LM or piecemeal resection. Among 3,782 EGC lesions (3,316 Pregnenolone patients) treated with ESD at our institution between 1997 and 2010, 85 non-curative differentiated-type EGC lesions (83 patients) were included in this study which met both of the following criteria: i) non-curative factor limited to a positive LM or piecemeal resection, and ii) follow-up period > 1 year. These patients underwent gastrectomy, immediate additional endoscopic treatment, or close observation after ESD. Close observation was performed only when no residual tumor was found endoscopically just after

ESD, and involved endoscopic follow-up every 3 to 6 months with computed tomography as needed for the first 3 years, followed by annual follow-ups. We retrospectively evaluated the occurrence of residual/locally recurrent cancer, metastasis, metachronous gastric cancer and death. To identify risk factors for residual/recurrent cancer among various clinicopathological features (see Table 1), univariate and multivariate logistic regression analyses were performed. The clinical course is summarized in Figure 1. The median follow-up period was 50 months (range 13-163). A total of 16 of the 85 lesions (18.8%) had residual/locally recurrent cancer; no metastasis occurred. The median period for local recurrence after ESD was 13 months (range 4-89). All residual/locally recurrent lesions were mucosal differentiated-type adenocarcinoma. There were 12 cases of metachronous gastric cancer. The 5-year disease-specific and overall survival rates were 100% and 92.1%, respectively.

The time for maximum facilitation after the return to 37 °C was 7.6 ± 0.3 min (n = 4) compared to 30 ± 5.8 min (n = 3) without prior incubation at 22 °C (p < 0.05); similarly,

the return to basal values was faster after pre-incubation at 22 °C compared to no pre-incubation (60 ± 12.3 min vs. 96.7 ± 12 min, respectively; p < 0.05); however, there was no difference in the maximum facilitation seen at 37 °C with or without pre-incubation at 22 °C (overall increase in tension of 106 ± 17% vs. 110 ± 12%, respectively). Venom PLA2 activity decreased by 91.4% at 22 °C compared to 37 °C (from 8.2 ± 1.3 U/mg to 0.7 ± 0.03 U/mg; n = 4). Incubation with BPB inhibited venom PLA2 activity by 89% and markedly attenuated venom (0.3 μg/ml)-induced neuromuscular blockade in chick biventer cervicis preparations (Fig. 1A); this inhibition also retarded the facilitation and attenuated the blockade by 30 μg selleck of venom/ml in mouse phrenic nerve preparations, without affecting the maximum facilitation observed (Fig. 2C) (the slower initial rise in facilitation in the presence of BPB-inhibited PLA2 probably

reflected the attenuated release of presynaptic ACh, as did Gefitinib nmr the attenuation of neuromuscular blockade from 60 min onwards). The finding that the inhibition of PLA2 activity delayed the onset but did not attenuate the maximum facilitation caused by the venom suggested that at least two components are involved in the neuromuscular

responses to venom in PND preparations, i.e., one that causes prolonged facilitation (non-PLA2) and one that contributes partially to the initial phase of facilitation and causes neuromuscular TCL blockade (most likely PLA2). To investigate this possibility, we examined the responses to venom in directly stimulated curarized PND preparations (to prevent the effects of presynaptically-released ACh). Fig. 2D shows that the venom indeed had a direct facilitatory effect on striated muscle that was independent of the neuromuscular blocking activity. Note that the time-scale and profile of this facilitatory response were very similar to those seen with BPB-treated (PLA2-inhibited) venom (Fig. 2C). The results described here show that B. b. smargadina venom causes potent neuromuscular blockade in avian and mammalian preparations in vitro, with avian preparations being ∼10 times more sensitive than mammalian preparations. This finding agrees with studies showing that Bothrops venoms and their basic PLA2 can cause neuromuscular blockade in vitro ( Zamunér et al., 2004 and Gallacci and Cavalcante, 2010). Although classic α-neurotoxins (nicotinic receptor antagonists) have not been identified in these venoms, various studies have shown that the venoms of some Bothrops species, e.g., B. insularis ( Cogo et al., 1993), B. pauloensis ( Borja-Oliveira et al., 2003 and Rodrigues-Simioni et al.