1). Upon discovery, a limited amount (<1 h) of video observations (inset image, Fig. 1) were collected. Subsequent inquiry of the shipping company by NOAA revealed the container’s cargo to be 1159 steel-belted automobile tires. In January 2005, the NOAA Damage Assessment Center (DAC) assessed the prospective financial impact of the deposition and deterioration of the 15 containers lost in the MBNMS. With consideration of NOAA-DAC’s evaluation, as well as potential fines, legal fees and costs to date, etc., the shipping company paid the MBNMS reparation of $3.25 million. The Compensatory Restoration Plan implemented by the MBNMS

includes assessment and monitoring

of the deep-sea benthos check details at the container site. The site was revisited for this purpose during a March 2011 research cruise as a collaborative venture between MBNMS and MBARI scientists. The aim of this cruise was to produce a detailed assessment of the diversity, abundance, and assemblages of benthic mega- and macrofauna on and around this intermodal container, seven years after its deposition in the MBNMS. Habitat heterogeneity increases biodiversity (Buhl-Mortensen et al., 2010, Levin et al., 2010 and Ramirez-Llodra et al., 2011), with natural click here and artificial structures typically attracting high densities and a

wide variety of marine taxa; so long as structures are not made from materials acutely toxic to prospective inhabitants (Bohnsack and Sutherland, 1985, Baine, 2001 and Collins et al., 2002). Indeed, artificial reefs are frequently installed in coastal regions at depths <100 m to enhance the diversity and abundance of ecologically and commercially important marine species (Bohnsack and Sutherland, 1985 and Baine, 2001). Artificial reefs have been shown to affect biological productivity and ecological connectivity; however, the types of organisms and their Ponatinib supplier persistence on and around a newly introduced structure depend largely on their shape, composition, and location (Bohnsack and Sutherland, 1985, Baine, 2001 and Macreadie et al., 2011). Although there is general scientific agreement that artificial reefs accumulate fish and other organisms (Bohnsack and Sutherland 1985), less is known about the effects of artificial reefs on living resource production, their ability to act as stepping-stones that facilitate the dispersal of native and non-native species, how they affect disease frequency in fish and invertebrates, toxicological impacts, their long-term structural integrity, and changes to socioeconomic conditions of adjacent coastal communities (Broughton 2012).

Settlement plates can be deployed to assess whether the colonising community has the same species composition as the previous community and/or

set aside area. Genetic analysis comparing the fauna colonising artificial or newly-generated natural substrate to the original populations could enable the source of colonisers to be identified drug discovery and the suitability of set aside areas to be assessed. The monitoring program needs to be implemented at suitable spatial and temporal scales (IMMS, 2011), although the appropriate length of long-term study required is at present unclear. Levels of natural variation need to be evaluated before any appreciable operations begin, in order to establish fluctuations that could, for example, be seasonal or related to changing chemical conditions. Also, following disturbance, succession of species composition and abundance is to be expected, and so any monitoring must span sufficient time. Recovery from natural disturbance at sites along the EPR (Lutz et al.,

1994 and Mullineaux et al., 2010) and Juan de Fuca Ridge (Tunnicliffe et al., 1997) and the rapid re-growth of deposits at Solwara 1 (Gwyther, 2008a) indicate that monitoring for a few years following the cessation of mining activities may be sufficient. However, experimental polymetallic nodule mining resulted in Tariquidar disturbance to the benthic community assemblage for at least 26 years following mining activity (Miljutin et al., 2011), suggesting that in keeping with the precautionary principle, suitable long-term monitoring could be on the scale of decades rather than years. Monitoring programmes by themselves are all very well, but they need to be evaluated against pre-determined decision rules. The latter will be derived from management objectives, and involve a management response when a monitored parameter value exceeds a certain level. For example, mining may have to stop in an area if sediment plume deposition thicknesses exceed a certain ZD1839 cell line depth. The design of baseline, impact and long-term monitoring studies also needs to consider the importance of replication to address the natural

environmental variability at SMS sites at both temporal and spatial scales. Ideally, this should utilise a design similar to BACI (before-after-control-impact, Green (1979)) or Beyond BACI (Underwood, 1991 and Underwood, 1992), with multiple unimpacted (control or set aside) and impacted (mined) sites (Collins et al., 2013a). However, BACI design at SMS sites will probably be asymmetrical with the potential for multiple unimpacted sites but only one impacted site (Underwood, 1991 and Underwood, 1992), as mining is likely to be concentrated at one site. There is also the question of cost. Coastal or shallow water impact studies may be able to investigate multiple sites but the logistics (time and cost) of investigating multiple sites in deep-sea SMS mining impact studies may be prohibitive.

, 1991 and Tallal et al., 1994). Although the correspondence between the two sets of studies is impressive the pattern of abnormalities in SLI also differs from that seen in the KE family

in several ways. In the current study, grey matter in the posterior temporal cortex in SLI is significantly find more decreased relative to controls, whereas it was increased in affected KE family members. Similarly, the putamen was found to have increased grey matter in affected KE family members, whereas we found no structural differences in the putamen in SLI. Finally, the caudate nucleus was found to be significantly reduced in volume in affected KE family members relative to their unaffected relatives and functionally overactive in a PET study of word repetition (Watkins et al., 1999). In our SLI study and the functional MRI study of the KE family, the caudate nucleus was not reliably activated by the task used and no group differences in function were detected. Also, the unaffected siblings in our study had significantly less grey matter in the caudate nucleus bilaterally relative to the typically developing controls

and did not differ significantly from their siblings with SLI. The latter suggests that reduced caudate nucleus volume can be considered a heritable risk factor for SLI but requires additional deficits selleck to affect language development. Alternatively, the siblings in our study have some protective factors, plasticity or compensatory mechanisms available to them that are unavailable to their affected

siblings. The increased grey Low-density-lipoprotein receptor kinase matter of the left central opercular cortex in the unaffected siblings relative to the SLI and control groups might reflect such compensatory mechanisms. The similarities between the functional and structural abnormalities in this group of people with SLI and the affected members of the KE family are likely a reflection of the similarities in their behavioural deficits. Both groups have impairments in nonword repetition and oromotor function. Whether any of the individuals with SLI that we studied also have a mutation in FOXP2 is unknown, but is unlikely, given the rarity of such mutations in individuals with SLI ( Newbury & Monaco, 2010). In a larger population of SLI, however, allelic variation in a downstream target gene of FOXP2, CNTNAP2 was found to correlate with performance on nonword repetition ( Vernes et al., 2008), so investigations of this gene in our participants are warranted. Previous developmental studies measuring grey matter volume and cortical thickness have revealed gradual linear and nonlinear reductions in these measures that continue into the second decade (e.g., Giedd et al., 1999, Giorgio et al., 2010 and Gogtay et al., 2004).

Com efeito, é necessário ter presente que até 20% dos doentes com história de abuso de álcool apresentam DNA Synthesis inhibitor uma causa secundária ou coexistente de doença hepática 26. O diagnóstico histológico de esteato-hepatite alcoólica baseia-se no achado de fígado gordo com um quadro de esteatose predominantemente macrovesicular, acompanhado de infiltrado inflamatório e lesão hepatocitária. O infiltrado inflamatório está

geralmente presente em focos lobulares dispersos, podendo atingir os espaços porta, constituído por neutrófilos, linfócitos, plasmócitos e macrófagos. A lesão hepatocitária mais frequente é a degenerescência em balão ou balonização dos hepatócitos. Normalmente, é mais proeminente na zona 3, onde se pode associar a fibrose perissinusoidal e outros hepatócitos esteatósicos. Outro achado histológico comum são os corpos de Mallory-Denk. A presença de colestase canalicular, proliferação ductular, lesões veno-oclusivas e Navitoclax nmr necrose hialina esclerosante é muito sugestiva da etiologia alcoólica da esteato-hepatite28.

Recentemente, foi relatada a utilidade de usar um corante imuno-histoquímico para K8/18, com vantagem de uma maior uniformização na interpretação das biopsias hepáticas para avaliar a gravidade da HAA, podendo produzir informação diagnóstica e prognóstica relevante29. A biopsia tem também a vantagem de permitir um estadiamento muito mais preciso da DHA. A ecografia hepática deve ser efetuada em todos os doentes com suspeita de HAA. É útil para excluir obstruções das vias biliares,

abcessos hepáticos e carcinoma hepatocelular, no diagnóstico diferencial da icterícia7 and 21. Masitinib (AB1010) Foi também demonstrado que a HAA está associada com um aumento do diâmetro e do fluxo da artéria hepática, que pode ser medido através do modo doppler duplex30. A elastografia hepática transitória é um avaliador não invasivo da fibrose hepática. É efetuada com um transdutor de ultrassons, que, baseado na elastografia transitória unidimensional, consegue medir a velocidade de propagação, que está diretamente relacionada com a elasticidade hepática. É um método rápido, indolor, reprodutível e pouco dependente do operador31. Contudo, a elastografia pode não ser adequada na presença de esteato-hepatite, sobrestimando a presença de fibrose32 and 33. A tomografia axial computadorizada abdominal não é usada por rotina no diagnóstico da HAA. Não existem dados sobre nenhum tipo de imagem característica da HAA, usando este método de imagem, como também pode ser um fator de confusão ao revelar lesões pseudotumorais na forma de áreas com hipervascularização arterial, que poderão corresponder a focos de intensa hiperplasia regenerativa focal34. Alguns centros diferenciados efetuam a medição do gradiente de pressão venoso hepático, cujo aumento está associado a uma maior mortalidade35.

In contrast, 5 patients with mutant cfDNA had no corresponding mutations in matched tumor tissue. This phenomenon has also been reported and could

be explained by tumor heterogeneity: these biopsied tumor tissue samples may not carry the EGFR mutations detected in blood, because these mutations come from different parts of the tumor [25], [26] and [27]. However, 4 of these 5 patients received EGFR-TKIs and had a comparable PFS with those who exhibited PS-341 wild type in both blood and tumor tissue, suggesting that these mutations detected in blood could be false positive results. There have been a limited number of studies on the correlation between EGFR mutation status in cfDNA and efficacy of EGFR-TKIs [28], [29], [30], [31] and [32]. Though the researchers tend to agree that EGFR activating mutations in cfDNA may be predictive of better response to EGFR-TKIs, they are still uncertain whether EGFR mutation status in cfDNA can predict survival benefit from EGFR-TKIs. In a subgroup analysis of IPASS, ORR was 75.0% (18/24) and 27.1% (19/70) with gefitinib in patients with or without EGFR mutant cfDNA, respectively.

PFS was significantly longer with INCB024360 order gefitinib than carboplatin/paclitaxel in the cfDNA mutant subgroup (hazard ratio [HR], 0.29; 95% CI, 0.14-0.60; P < 0.001) but not in the cfDNA wild-type subgroup (HR, 0.88; 95% CI, 0.61-1.28; P = 0.50) [22]. Xu et al. reported that an significant correlation between EGFR mutations status in plasma and tumor response to gefitinib was observed using ARMS but not denaturing high-performance liquid chromatography (DHPLC), whereas no association between EGFR mutation status

in plasma and PFS or overall survival (OS) was observed no matter using ARMS or DHPLC [33]. Bai et al. detected EGFR mutations in plasma using DHPLC and found that about 62.2% of patients with EGFR mutations responded to gefitinib, whereas 37.8% of patients with wild-type EGFR also responded. They noted that patients with EGFR mutant cfDNA had a significantly next longer PFS than those with wild-type cfDNA (11.1 months versus 5.9 months, P = 0.044), though no difference in OS was seen [25]. In the current study, patients with EGFR activating mutations in tumor tissue had significantly greater ORR and longer PFS with EGFR-TKIs, which accords with the finding of previous clinical trials [4], [5], [6], [7] and [8]. Patients harboring EGFR activating mutations in cfDNA also had significantly higher ORR, which was consistent to that of patients with mutant tumors. In addition, patients with mutant cfDNA tended to have longer PFS than those with wild-type cfDNA, though the difference was not significant. These data suggest that EGFR activating mutations detected in blood may be predictive of improved tumor response and survival benefit from EGFR-TKIs.

2 °C min−1), to http://www.selleckchem.com/products/dinaciclib-sch727965.html (i) progressively lower sub-zero temperatures (−12.5 to −19.5 °C) below the DTemp for 8 h, before re-warming to +4 °C at 0.2 °C min−1, and (ii) progressively longer periods (10–48 h) at the DTemp, before re-warming to +4 °C at 0.2 °C min−1. Soil temperature data available from previous seasons at Signy Island and Anchorage Island (67oS

68oW) were used as a basis to establish two thermoperiods; one that E. murphyi currently experiences in summer on Signy Island, and one that might be experienced in summer on Anchorage Island. This was undertaken to assess the ability of E. murphyi larvae to survive at a more extreme, higher latitude, location. Using these models, an alcohol bath was programmed to cycle between +6 and −1 °C, and between +4 and −3 °C, representing Signy and Anchorage Islands respectively,

over a 24 h period ( Fig. 1). Larvae were transferred to each thermoperiod (beginning at 4 °C). Three replicates of 10 individuals were removed at two points in the cycle (−1 and 6 °C [Signy Island model] and −3 and 4 °C [Anchorage Island model]) each day for 3 days during each thermoperiodic cycle and directly transferred to the DTemp for 8 h, before being re-warmed to +4 °C at 0.2 °C min−1. To determine the effect of RCH on the SCP, juvenile and mature larvae were cooled from +4 to −30 °C at either 0.2 °C min−1 (RCH treatment) or 1 °C min−1 (mature larvae only). Controls were directly transferred to the DTemp. Juvenile and mature larvae (8 and 24 individuals) were placed in contact with a thermocouple, within Beem capsules, in glass BMN 673 mw test tubes plugged with sponge, inside an alcohol bath, prior to each cooling regime. SCPs, defined as the temperature at the

onset of the freezing exotherm, were identified using an eight channel datalogger interfaced to a computer and recorded using PicoLog Recorder Software (Pico Technology Limited, UK) (cf. Hawes et al., 2006). The time at which all mature larvae froze at −7 °C, having been cooled at 1 °C min−1 from +4 °C, was calculated as 4 min using PicoLog Recorder Software (Pico Technology Limited, UK). Three groups of 10 mature larvae were subsequently cooled from +4 to −7 °C at 1 °C min−1, held for 4 min or 1 h 4 min, and transferred to the DTemp for 8 h, before being re-warmed Tyrosine-protein kinase BLK to +4 °C at 0.2 °C min−1. Survival was assessed 24 and 72 h after each treatment. The Kolmogorov–Smirnov test was used to confirm that all percentage survival and SCP data were normally distributed. The data were subsequently analyzed using analysis of variance (ANOVA) and Tukey’s multiple range test. The mean survival of both juvenile and mature larvae decreased significantly following exposure to progressively lower sub-zero temperatures for 8 h (Fig. 2; P < 0.05 Tukey’s multiple range test), declining from more than 80% at −9 °C to 0% at −14 °C.

Suspicious aspects include a large nodule, depression and loss of pit pattern, and a masslike appearance (Fig. 2).13 The presence of any of these signs should lead to a careful consideration of whether endoscopic resection is appropriate. Unfortunately, these techniques, which are reasonably reliable in noncolitic colons, perform less well in colitis, because the scarring may lead to pseudodepression and inflammation distorts pit patterns. The nonlifting sign, which in combination with macroscopic

appearance gives a good estimate of likely invasion in the assessment of noncolitis-associated lesions, is by definition poor in colitis. Submucosal scarring impedes mucosal lift14 and also disrupts the mucosal layers needed

to clearly assess invasion at endoscopic ultrasonography. In noncolitis cases, submucosal scarring can LDN-193189 cell line be seen in lesions with a previous attempt at resection, recurrence on a scar from previous EMR, or nongranular type LSTs.15 In colitis cases, if the patient has a tubular colon with evidence of scarring, postinflammatory polyps, loss of vascular pattern, or active inflammation, the submucosal scarring is likely to be severe and typically involves the entire lesion. Location of the lesion near technically difficult areas such as the appendix orifice, ileocecal valve, at a flexure, especially on the inside of the bend, and at the anal verge should also be considered.16 Although polyps Crenolanib in all these positions can be resected in noncolitic colon by experienced endoscopists, the technical difficulty is substantially increased. In combination with the other inherent challenges that colitic lesions present, this may make the likelihood of a successful resection so low that an Erastin endoscopic attempt is not appropriate. The final stage is to consider endoscopic access. This is one of the few areas in which working in a colitic colon may have advantages because a scarred and tubular colon makes for a straight endoscope and associated

accurate tip movements and a lack of haustral folds to be negotiated. Before starting, endoscopists should be satisfied that they can easily reach all areas of the lesion with submillimeter precision. There is no specific combination of factors or scoring system that suggests that lesions are or are not safely and effectively resectable. Ultimately, at least at present, it comes down to the experience and judgment of the assessing endoscopist. Given the fine nature of these judgments, the authors recommend that if possible the endoscopist who is going to do the resection procedure should perform the endoscopy for lesion assessment before resection. Lifting or the failure of lifting of lesions in colitis is one of the major obstacles to resection.

From −110 to +10 mV we observed the

normal decrease of the Af and As components, but also the increase of the Ass component as shown in the inset to Fig. 2 upper-right (see under Nav1.1 panel). The steady-state inactivation resulted to have a sigmoidal voltage-dependent curve, which, differently from control curves, was characterized by a KU 57788 non complete inactivation and a pedestal at depolarized potentials. This last Ass effect indirectly and strongly affected the window current (normally negligible in control), thus producing a small and not always significant left-shift of the activation curves. When necessary the dose-response relationships of As and Ass components were computed in Fig. 4. In order to visualize the 3D structure of each studied toxin, models of CGTX-II, δ-AITX-Bcg1a and δ-AITX-Bcg1b were constructed using the SWISS-MODEL structure homology-modeling server (http://swissmodel.expasy.org/workspace/) [3]. The tri-dimensional structure of Anthopleurin-A toxin (determined by NMR) [24] (PDB ID: 1ahl) was employed as a template for all models.

The structures were find more drawn and visualized by DeepView/PDB viewer [12] (http://www.expasy.org/spdbv, version 4.0.1) and PyMOL (The PyMOL Molecular Graphics System, Version 1.2, Schrödinger, LLC., http://www.pymol.org), and were rendered by PovRay (version 3.6 by Persistence of Vision Raytracer, Pty., Ltd.). All the three models were validated by the tools Anolea, DFire, QMEAN, Gromos, Promotif and ProCheck, available in the “structure assessment” tool of the SWISS-MODEL structure homology-modeling server. The toxins employed in this study were obtained according to the previously described procedures [35] and [36]. Considering an urgent need for standardization of nomenclature of animal toxins, especially in sea anemones, we employed a rationale recently suggested [17]

for the novel components eluted during RP-HPLC at 30.24 and 30.57 min from the neurotoxic fraction of B. cangicum venom. Decitabine These peptides were named as δ-Actitoxin-Bcg1a (δ-AITX-Bcg1a) and δ-Actitoxin-Bcg1b (δ-AITX-Bcg1b), respectively. This rationale follows the biological effects exerted by the toxin (δ letter for toxins that delay the inactivation process of ion channels) and the family of the organism which the toxin is derived (“Actitoxin” for toxins isolated from sea anemones of the “Actiniidae” family). Also, full sequences of each peptide were determined in this work. Both sequences were deposited at Uniprot server (http://www.ebi.ac.uk/uniprot/) and their accession numbers were assigned as P86459 and P86460, respectively. As CGTX-II (Uniprot ID: P0C7P9) had already been published [35], we have not changed its name.

The mTOR inhibitor first-pass assembly was performed with Velvet and ABySS to assemble short contigs using both our Illumina reads and available ESTs ( Simpson et al., 2009 and Zerbino

and Birney, 2008). Second-pass assembly was completed with the MIRA assembler to combine Velvet and ABySS contigs, steelhead Illumina reads, and tentative consensus transcripts into longer transcript scaffolds ( Chevreux et al., 1999). A total of 86,157 transcript scaffolds were greater than 100 bp with an average length of 902 bp (Fig. 1; Supplementary file 1). We assembled 27,095 transcript scaffolds greater than 1 kb averaging 1509 bp. Of the 86,402 transcript scaffolds, 49,149 homologous genes were identified using E-value cutoff of 10− 5. We assigned GO terms to each sequence using the Blast2GO tools (Supplementary file 2; Gotz et al., 2008). A total of 4030 gene ontology definitions were identified among the group of 24,624 transcript scaffolds. The biological processes class was the most highly Sotrastaurin mw represented (68.3%), followed by molecular function (22.2%) and cellular component (9.5%; Fig. 1). See Supplementary Methods for more details regarding functional annotation. The top BLASTx hits to other fishes included

zebrafish (Danio rerio), Atlantic salmon (Salmo salar), and pufferfish (Tetraodon nigroviridis and Takifugu rubripes) ( Fig. 2). In addition, we conducted comparisons between the rainbow trout/steelhead assemblies and zebrafish proteins ( Fig. 2;

See Supplementary Methods for details). The O. mykiss raw sequence data from this study is accessioned in the NCBI Sequence Read Archive (SRA accession SRP009644.1). We have also made a searchable BLAST database to facilitate research using aminophylline steelhead trout (http://salmon.cgrb.oregonstate.edu/). All transcriptome reference contigs, predicted proteins, and associated GO terms are available via FTP on the Salmon Resource website. The following are the supplementary data related to this article. Supplementary Methods. We thank the Oregon Department of Fish and Wildlife for their assistance in collecting the hatchery and wild steelhead trout, and staff of the Parkdale and Oak Springs hatchery for crossing and raising the fish, in particular, Jim Gidley and Lyle Curtis. We are grateful to Anne-Marie Girard and Caprice Rosato for the qualitative assessment of RNA and cDNA and Mark Dasenko (Center for Gene Research and Biocomputing, Oregon State University) for Illumina cluster generation and sequencing. We thank Matthew Peterson and especially Chris Sullivan (Center for Gene Research and Biocomputing, Oregon State University) for computational support. “
“By virtue of their abundance, surface area and metabolic versatility, microbes mediate the vast majority of organic matter transformations in the ocean and control the flow of energy and key nutrients (C, N, P, S, Fe) to higher trophic levels (Falkowski et al., 2008).

1B). The incubation of mouse diaphragm muscle with P2 (30 μg/ml) resulted in a significant decrease in quantal content from 15 min onwards [from 70 ± 6 (basal) to 19 ± 3 after 60 min; n = 5; p < 0.05] ( Fig. 1C) and in the frequency of MEPPs from 5 min onwards [from 33 ± 3 (basal) to 16 ± 1 after 60 min; n = 5; p < 0.05] ( Fig. 2A). There was no change in MEPPs amplitude in PND preparations treated with

P2 (30 μg/ml) (0.8 ± 0.06 mV at t0 compared to 0.8 ± 0.04 mV at t60). In contrast to P2, P3 (30 μg/ml) produced an increase in quantal content from 5 min onwards although statistical significance BMS-907351 cost was seen only after 30 min [from 63 ± 6 (basal) to 85 ± 6 after 60 min; n = 5; p < 0.05] ( Fig. 1D). At this same concentration, P3 also increased the frequency of MEPPs from t30 onwards [from 20 ± 2 (basal) to 30 ± 3 after 60 min; n = 5; p < 0.05] ( Fig. 2B) without altering their amplitude (1.0 ± 0.2 mV

at t0 compared to 0.8 ± 0.1 mV at t60). Bbil-TX (0.5–10 μg/ml) produced irreversible time- and concentration-dependent neuromuscular blockade in indirectly stimulated BC preparations, with complete blockade occurring after 41 ± 2 min (n = 6) at the highest concentration (10 μg/ml); Bbil-TX reproduced the neuromuscular blockade seen with peak P2 (10 μg/ml) from which the toxin was purified ( Fig. 3A). The times required for 50% blockade were 87 ± 7, 41 ± 7 and 19 ± 2 min for Bbil-TX concentrations of 1, 5 and 10 μg/ml, respectively; the time required for 90% blockade was 37 ± 2 min for the highest Bbil-TX concentration. Fig. 3B1 (upper trace) shows a representative Carfilzomib order recording of the neuromuscular blockade produced by Bbil-TX (10 μg/ml) under indirect stimulation at 37 °C. There were no consistently significant changes in the contractures to exogenous ACh and KCl after complete neuromuscular blockade by Bbil-TX ( Fig. 3C). When the experiments were done at 22–24 °C Bbil-TX (5 μg/ml) caused only 21 ± 5% blockade

after 120 min whereas complete blockade was seen at 37 °C after 90 min (Fig. 3A). Pretreatment Cobimetinib price of Bbil-TX with p-BPB abolished the PLA2 activity (88 ± 6 units/mg vs. 2 ± 2 units/mg before and after p-BPB, respectively; n = 3) and also abolished the neuromuscular blockade by this toxin ( Fig. 3A; responses superposed on control preparations). The neuromuscular blockade normally caused by Bbil-TX (10 μg/ml) was absent in curarized (d-Tc, 10 μg/ml) directly stimulated BC preparations, with the twitch-tension response being similar to that of control preparations (Fig. 3B2, D). Bbil-TX caused partial time- and concentration-dependent neuromuscular blockade in indirectly stimulated PND preparations (maximum blockade of 15.2 ± 3%, 29.8 ± 3% and 52.2 ± 2% of the control for concentrations of 3, 10 and 30 μg/ml, respectively, after 120 min; n = 4–6).