Nevertheless, this hypothesis has been challenged by other studies suggesting that tourism activities stimulate deforestation and forest degradation. Research by Forsyth (1995) in northern Thailand showed that the growth of the tourism sector did not decrease agricultural pressure on forests and soil resources because households invested their income from tourism in the expansion of arable fields and increasing frequency of cultivation by hiring external Smoothened inhibitor labour. Additionally, Gaughan et al. (2009) showed that the increased number of visitors to the archaeological sites of Angkor Kwat in Cambodia accelerated deforestation in the Angkor

basin. The deforestation occurred due to increased charcoal production for new restaurants and hotels, which required wood products from forests. In the coastal areas of Hainan Island (Southern China) and the Mediterranean (Turkey), Wang and Liu (2013) and Atik et al. (2010) respectively indicated that tourism development led to a rapid increase of the built-up area. These activities resulted in a decrease of agricultural land and coastal forest, causing

landscape fragmentation and coastal erosion. In this study, we evaluate possible changes in the human–environment interactions after the development of tourism activities. Using Sa Pa district in the northern Vietnamese Highlands as a test case, we addressed the following questions: First, how has forest cover changed in http://www.selleckchem.com/products/bgj398-nvp-bgj398.html the period between 1993 and 2014? Second, how does forest cover change relate to tourism development? Third, what are the likely impacts of the changing human–landscape relationships on local livelihoods? Sa Pa district is located in Northern Vietnam (Fig. 1) and covers an area of ca. 680 km2. It has a total of 55,900 inhabitants (GSO, 2010) living in 17 communes and its administrative centre, Sa Pa town.

The district is considered as a gateway to the northern Vietnamese Highlands. The topography is rough, with an elevation of 180 m in the Muong Hoa valley and up to 3143 m at the Fansipan peak (highest elevation in Vietnam, located within Hoang Lien National Park). The major rivers are the Muong Hoa and Ta Trung Ho River that flow in the Red River nearby 4-Aminobutyrate aminotransferase Lao Cai. The region is characterized by a sub-tropical and temperate climate with an annual rainfall of 2763 mm (Frontier Vietnam, 1999). Sa Pa district is home to 6 major ethnic groups: the Hmong, the Yao, the Tày, the Giáy, the Xa Pho and the Kinh. The Tày occupied the fertile valleys and middle altitudes. The other ethnic groups such as the Hmong and Yao entered Northern Vietnam only in the 19th century (Michaud and Turner, 2006), and settled on steep forested slopes generally above 800 m. Before 1960s, there were only a few Kinh lowlanders living in Sa Pa town as the surveillance and maintenance staffs of French military (Michaud and Turner, 2006).

5, Varian Inc., Palo Alto, CA, USA) and a 30 m fused silica capillary column (ID = 0.25 mm) coated with 0.25 mm of CP-Wax 52CB (Chrompack, Minnesota, MN, USA). Helium carrier gas flow was 1.5 ml/min at a split ratio of 1:50. Injector temperature was 250 °C. Detector temperature was 280 °C. The oven temperature was set initially at 75 °C for 3 min, and then programmed at 37.5 °C/min to 150 °C and at 3 °C/min to 215 °C. After drawing up some air into the filled syringe (sample volume 1 μl) and inserting the needle into the heated injector, samples were injected manually after a dwell-time of 2 s. Qualitative FA composition was determined by comparing

the retention times of the peaks with the respective FA standards. Quantitative composition was accomplished OSI-906 in vitro by area normalization. The proportion of each individual FA (FAi) in the whole

samples was estimated according to the Equation (1): equation(1) FAi(g/100g)=FAMEi×(FAiMWFAMEiMW)×FAT×0.933∑FAMEi×(FAiMWFAMEiMW),where Selleckchem Vorinostat FAMEi is the percentage of each individual FAME (g/100 g total FAME), FAiMW and FAMEiMW are the corresponding FA and FAME individual molecular weights, FAT is the percentage of total fat in samples (g/100 g) and 0.933 is the coefficient for the mean FA proportion in total milk fat described by Glasser, Doreau, Ferlay, and Chilliard (2007). Protein was analyzed by measuring the nitrogen content of mousses through the micro Kjeldahl method and multiplying by a conversion factor (6.38), according to the AOAC official methods 690.52 and 991.20 (AOAC, 2003). Dietary fibre other than fructans (DFotf) estimates were adapted from AOAC method 985.29 for total dietary fibre (TDF) (Prosky et al., 1985). The modification lies

in an additional enzymatic treatment of mousse samples with a purified fructanase mixture E-FRMXLQ (2000 units exo-inulinase and 200 units endo-inulinase per ml, Megazyme, Bray County, Ireland, 1 ml/g fructan, 60 °C, 30 min) for the complete hydrolysis of fructans, once these types of dietary fibres are not totally quantified through the original method, which leads to an underestimate of the real content of TDF. This analysis was carried out on duplicate samples. The fructan content of samples was estimated based on the fructan present in whole Beneo P95 and Beneo Farnesyltransferase HP-Gel batches used for the addition in the mousse trials, 95.2 g/100 g and 98.5 g/100 g, respectively (data given by Orafti). The DFotf plus the fructan values from those ingredients were used to estimate the TDF content of samples. Available carbohydrate content (carbohydrate excluding TDF), was obtained by difference in order to achieve 100 g/100 g of total composition (FAO, 2003). Total energy value from each mousse formulation was obtained from energy equivalents for available carbohydrate, fat, and protein, 4 kcal/g, 9 kcal/g, and 4 kcal/g, respectively (FAO, 2003), and mean energy from inulin and FOS (1.5 kcal/g) (Roberfroid, 1999).

Finally,

1:1 phase synchrony between the resultant filtered envelope and the original envelope of the slower oscillation was estimated using PLV1:1 statistics to quantify the strength of phase-amplitude modulation (referred to as PLVPAM), which is one of the most common manifestations of nesting. The analysis of spike timing with respect to LFP phase was performed for gamma, alpha and theta oscillations during an active attractor-coding state. The reference rhythms and their instantaneous phase were obtained by filtering and applying a Hilbert transform. In addition, a more detailed examination was made to distinguish peaks in the phase distribution Dolutegravir concentration for the gamma rhythm when studying individual Lumacaftor research buy contributions from basket and pyramidal cells. The signals generated within each hypercolumn were then matched with the spikes produced by the corresponding

local basket and pyramidal cells. In addition, spikes produced by basket cells from other hypercolumns were also examined to compare with the local phase distribution. All the firing phase distributions were statistically assessed using circular statistics. First, in order to test the null hypothesis about uniform circular distribution of the gamma phases of spikes produced locally by pyramidal and basket cells, Rao’s spacing test (Rao, 1976), Hodges–Ajne test (Zar, 1999) and the standard Rayleigh test (Fischer, 1995) were performed (Berens, 2009). Since the hypothesis about the uniformity of the phase firing was rejected in both cases at the 0.05 significance level, it was verified whether these circular random variables followed a von Mises distribution by estimating Watson’s U2 statistic ( Lockhart and Stephens, 1985). However, the hypotheses for both pyramidal and basket cells were rejected and in consequence, a nonparametric evaluation of the preferred phase of firing with 95% confidence intervals was performed using a bootstrap computation of the circular Hodges–Lehmann statistic as a point estimate along

with a so-called equal-tailed arc as an interval estimate ( Otieno, 2002 and Otieno Calpain and Anderson-Cook, 2006). These techniques have been demonstrated to perform well for skewed or nonsymmetrical sample distributions ( Otieno, 2002), as in the cases investigated here. Finally, the null-hypothesis that the two mean preferred phases (for pyramidal and basket cells) were equal was subjected to nonparametric permutation test (p<0.01). Circular visualization of firing phase distributions with respect to theta, alpha and gamma rhythms was made with the use of the circular statistics toolbox (Berens, 2009) in MATLAB. The analysis of instantaneous firing rates in the model during attractor activation was performed using peri-stimulus time histogram by aligning spike sequences with respect to the attractor onset.

For years, immunotoxin conjugates (e.g., HA-22) have been studied in patients with relapsed disease. In patients with resistant or relapsed hairy cell leukemia expressing CD22, the immunotoxin conjugate has been reported to result in complete responses in 46% of patients [62]. Many of these patients have had durable remissions with tolerable toxicities. The recent observation that patients with classic hairy cell leukemia carry BRAF p.V600E LY294002 molecular weight within their leukemic cells prompted the exploration of the BRAF inhibitor vemurafenib in the setting of relapsed and resistant disease [63]. In patients with well-documented failures to respond to multiple standard agents, this targeted

therapy has achieved durable remissions [64], [65] and [66]. Studies in both New York and Italy are formally exploring this agent in the setting of relapsed disease. Patients have been reported to show a rapid response to vemurafenib and other BRAF V600E inhibitors. The optimal therapeutic regimen with vemurafenib has yet to

be defined. Many of the current studies simply utilize the dose and schedule derived from other trials in patients with metastatic melanoma. Considering the cutaneous toxicities, there may be other doses and schedules of administration that would provide better outcomes in hairy cell leukemia. In patients who have relapsed, a search for pathways of resistance has suggested that MEK inhibition may be equally important. Tiacci and colleagues showed that the MEK–ERK pathway has

both diagnostic and therapeutic target potentials in hairy cell leukemia [67]. In a recent brief publication, the clinical Cabozantinib cost efficacy of vemurafenib was associated with BRAF inhibition, but surprisingly there was an uncoupling with phospho-ERK as measured in the in vivo leukemic cells obtained from the patient [68]. Thus, this intriguing pathway holds promise for therapeutic intervention while presenting ample opportunity for further investigation of pharmacodynamic effects. While BRAF inhibitors provide an option for patients with classic hairy cell leukemia in need of novel therapy, patients with the variant forms of this disease require other strategies. Patients with hairy cell leukemia variant do not respond well to standard purine analog treatment with low response rates and less durable remissions. They do not have BRAF mutations, eliminating Erastin both standard therapy as well as BRAF-targeted therapies. Patients with the variant may respond to HA-22, and responses have been observed with cladribine in combination with rituximab [17]. Recently, a multi-institutional trial involving the BTK inhibitor ibrutinib has begun to enroll patients with both the classic form in relapse and the variant of this disease (http://clinicaltrials.gov/ct2/show/NCT01841723). The necessity to explore novel therapies with tolerable side effects is highly warranted, as standard therapy is associated with toxicity and limited duration of response.

There are many mechanisms for achieving this, but the VVP program is extraordinarily effective and I am grateful to the Vallee Foundation for providing me with this opportunity. This says much about both the value of the scheme and, of course, the breadth of vision of Bert Vallee. Of course, the VVP scheme provided an excellent way of furthering long-standing interactions between a host institution and scientists. There had find more been a successful collaboration between Oxford and Gerard Canters from Leiden and when Bert agreed that he could be offered

a VVP, he, and his partner, were welcomed to Oxford. Gerard spells out the attractions to him succinctly: the excellent scientific reputation of the host institute; complete freedom of any bureaucratic obligations and the ability to focus on science and mutual contacts; the length of the stay. Longer than

three months a year would have caused problems in my own institution; shorter than a month would learn more have diminished the usefulness of my stay abroad disproportionately; the provision of adequate financial compensation. Needless to say, Gerard’s visit was incredibly valuable and the interactions started at that time still continue, indeed have expanded. The wish of Bert Vallee to make it easier for scientists to cross disciplines is well illustrated by Alan Bond’s appointment as a VVP. Alan has had a vast experience in electrochemistry both in its analytical use and in its application to organometallic chemistry. To strengthen his interest in its application to biochemical problems, he came to Oxford to work primarily with Fraser Armstrong “to further explore the mechanisms of catalytic processes associated with

cytochrome c peroxidase and on hydrogenases”. The analysis of the data obtained required the development of the second generation of Fourier Transform method of analysis. A wonderful aspect of the Vallee Foundation Progesterone has been facilitation of collaboration with experts from different fields. This has enabled stiff problems to be tackled at an in-depth level. To those who had known Bert Vallee for a long time, it was interesting to see the reaction of those who met him late in his life. This occurred for Professor Bond at his first Vallee meeting: It was an extraordinarily great pleasure to meet Bert Vallee in Boston and to see what he has achieved in his career and through the Foundation. Of course, there was special pleasure in having three VVPs from the Harvard Medical School: Peter Howley, Lew Cantley and Wade Harper. Peter spent his first week in Oxford at the DNA Tumour Virus Meeting, which included sessions on human papillomaviruses. It was a productive meeting and allowed Peter to discuss, with many of the other participants, the molecular mechanisms by which HPVs contribute to cervical cancer.

e., incomplete outcome data). In all cases, an answer of ‘yes’ indicates a low risk of bias, and an answer of ‘no’ indicates a high risk of bias [11]. Heterogeneity was quantified by χ2 and I2. The quantity, I2, describes the percentage of total variation across studies that is due to heterogeneity rather than to chance. Negative values of I2 are made equal to zero so that I2 selleck chemicals llc lies between 0% and 100%. A value of 0% indicates no observed heterogeneity,

and larger values show increasing heterogeneity. The results for individual studies and pooled statistics are reported as the risk ratio (RR) between the experimental and control groups with 95% confidence intervals (95% CI). The data were analyzed using RevMan. Fig. 1 shows the flow of studies through the selection process. A total of 714 records were identified from the primary electronic databases. Ten potentially relevant studies

BMS-777607 in vitro were identified for full-text review. Six RCTs met the inclusion criteria [12], [13], [14], [15], [16] and [17]. The characteristics of the included trials are presented in Table I. Excluded studies are described in Table II. The included RCTs randomized a total of 1343 patients (690 in the experimental group and 653 in the control group). Five included studies were double blind, placebo-controlled trials [13], [14], [15], [16] and [18]. All trials had some methodological limitations such as unclear allocation concealment [13], [14], [15] and [17] and/or no intention-to-treat

analysis [14] and [18]. Patients were hospitalized in pediatric departments for acute or chronic diseases. In the study by Saavedra et al. [18], children were admitted to a chronic medical care hospital. The most common reason for hospitalization was upper respiratory tract infection. One exception was the study by Hojsak et al. [13], in which children with respiratory tract infections were excluded, as this was one of the outcomes. Patients’ ages ranged from 1 month to 18 years. Five RCTs O-methylated flavonoid [14], [15], [16], [17] and [18] included only infants and young children under the age of 48 months. In contrast, in the study by Hojsak et al. [13], the mean age of the participants was 9.9 years, and children below the age of 12 months were excluded. Exclusion criteria for participants were mostly similar and included breastfeeding [15], [16] and [18], probiotic use within 7 days before admission [13], [15] and [16], acute gastroenteritis [13], [14], [15], [16], [17] and [18], gastroenteritis in the first 24 h after admission [15] and [17], and chronic gastrointestinal diseases [13], [15] and [16]. Only a limited number of probiotic microorganisms were tested. Three RCTs tested LGG [13], [14] and [15] at a daily dose ranging from 1 × 109 CFU [13] to 1 × 1010 CFU [14] to 6 × 109 CFU [15].

Additionally, it has been theorized that release of nitric oxide by nerves, vessels, or brain tissue may be part of the trigger of for migraine pain [79]. Hyperbaric oxygen causes cerebral vasoconstriction, likely though scavenging of nitric oxide [80] and thus the effect of HBO2T might improve pain directly through decreases in NO as well as through vasoconstriction and anti-inflammatory

mechanisms. There is some evidence that HBO2T is an effective treatment of acute migraine attack. Wilson et al. [81] assigned female subjects Nivolumab clinical trial with confirmed migraine to either 100% oxygen at normal pressures, or hyperbaric oxygen. They found that subjective pain was significantly reduced in the group receiving hyperbaric oxygen, but not following control treatment. They concluded that selleck HBO2T is effective for migraine pain, and the patient’s subjective pain assessment was the best indicator of relief. In a double blind, placebo-controlled

study by Eftedal et al. [82] the prophylactic effect of HBO2T on migraine was investigated. Forty patients were randomly assigned to a treatment group receiving three sessions of hyperbaric oxygen, or a control group receiving three hyperbaric air treatments. Patients kept a standardized migraine diary for eight weeks before and following treatments. Thirty-four patients completed the study. Their primary measure of efficacy was the difference between pre- and post-treatment hours of headache per week. The results showed a non- significant reduction in hours of headache between groups. Levels of endothelin-1 in venous blood pre- and post-treatment showed no difference between the hyperbaric oxygen and control groups. They concluded that the tested protocol does not show a significant prophylactic effect on migraine and does not influence the level of endothelin-1 in venous blood. Bennett et al. [83] conducted a meta-analysis on randomized trials comparing HBO2T or normobaric oxygen with placebo or no treatment in patients with migraine headache or cluster headache. Nine small Methane monooxygenase trials were included

which involved 201 participants. Five trials compared HBO2T vs sham therapy for migraine. Pooling data from three trials suggested that HBO2T was effective in relieving migraine headache compared to sham (relative risk (RR) 5.97, 95% confidence interval (CI) 1.46–24.38, P = 0.01). However, no evidence was found for prophylactic use. No reduction in the incidence of nausea and vomiting was seen. Neither was there a reduction in rescue medication requirements. We are not aware of data looking at HBO2T as a therapy for status migrainosus. Patients arriving to the Emergency Department with a presumed diagnosis of status migrainosus by history will be evaluated by a neurologist. Inclusion in the study requires that the patient, either male or female, be at least 18 years-old and have prior history of migraine consistent with current headache except in duration.

Moffat et BMS-754807 order al. (2007) have shown that liver miRNAs in both mouse and rat respond to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) treatment, a potent AHR agonist. However, the changes in expression levels for most of the miRNAs were modest and could not be confirmed by real-time RT-PCR, suggesting that hepatic miRNAs may play only a minimal role in AHR-mediated transcriptional hepatic response. Lastly, the gene expression responses in the liver and the lungs

of the BaP exposed mice were significantly different, with lungs exhibiting a pronounced immunosuppressive expression profile, in addition to xenobiotic metabolism, p53 signalling and oxidative stress. In conclusion, we have demonstrated a strong response in mRNA and miRNA in the lungs of

mice exposed to BaP by oral gavage. The pulmonary profiles of both gene and miRNA expressions resemble those of certain Galunisertib chemical structure types of lymphomas. The present study details the molecular mechanisms underlying BaP-induced immunotoxicity in lungs and its potential implications. Further research is needed to determine whether these molecular signatures can be used as markers for screening other environmental immunotoxicants, or if the miRNA expression profiles provide a better biomarker of immunotoxicity. The present study also highlights the importance of studying non-target organ toxicity in understanding the overall effects of a widespread chemical like BaP. There are no conflicts of interest to disclose. We thank Kelly Jackson for animal experimental design and sample collection, Lynn Berndt-Weis and Julie Buick for liver mRNA microarray data, and Karen

Leingardner for clinical chemistry. We also thank Christine Lemieux and David Lefebvre for helpful comments on the manuscript. “
“Hydroquinone (HQ) and benzene play an important role in both indoor and outdoor pollution as both are present Adenosine in high concentrations in cigarette smoke, where HQ is the most pro-oxidant compound (McGregor, 2007) and benzene is an environmental pollutant released from adulterated fuel. In addition, HQ is endogenously produced during benzene biotransformation, mainly in the lungs, liver and bone marrow, and it is also responsible for the myelotoxicity and immunosuppression detected during benzene toxicity (McGregor, 2007, Snyder, 2002 and Snyder, 2004). Cells and tissues present in the respiratory system are easy targets of the toxic actions of pollutants and pathogens dispersed in the atmosphere. In this context, a pool of active resident cells is necessary to provide a protective environment against inhaled microbes and to maintain host defence effectiveness (Soehnlein and Lindbom, 2010). The functional pool of alveolar macrophages (AMs) represents 90% of haematopoietic cells in the alveolar space and is responsible for eliminating invading agents, such as particles and microorganisms, by phagocytosis and killing activities (Geiser, 2002, Gordon and Read, 2002 and Laskin et al., 2001).

In those studies, filamentous algae, including Cladophora glomerata, Dictyosiphon foeniculaceus (Hudson) Greville and Ectocarpus siliculosus (Dillwyn) Lyngbye, were dominant at sheltered sites, whereas these species were present in only low biomasses during our spring study. C. glomerata possesses a number of traits that gives it a competitive advantage compared to other algae in shallow areas. It is promoted by higher temperature ( Snoeijs & Prentice 1989), it has a superior nutrient and carbon uptake capability ( Wallentinus, 1984 and Choo et al., 2005), as well as a better ability to cope with light stress in the upper littoral zone ( Choo et al. 2005). This is probably

the main reason for our contrasting results compared to the earlier studies, and the reason Bcl2 inhibitor OTX015 why we rejected our hypothesis that biomass would be higher at wave-sheltered sites. To describe the spring development in greater detail, the first species to exhibit increased biomass was the brown alga

P. littoralis. The explanation for the successful early establishment of P. littoralis is that it reproduces in winter ( Kiirikki & Lehvo 1997) and has the ability to grow rapidly at low temperatures (5 °C), compared to other competitive filamentous species like C. glomerata, D. foeniculaceus and E. siliculosus ( Lotze et al. 1999). The biomass produced by P. littoralis was substantially less than that found in the only other quantitative investigation conducted in the spring in the Baltic Sea: Kraufvelin et al. (2007) reported a 2 to 6 times higher Bacterial neuraminidase biomass of P. littoralis. This difference may be due to the higher nutrient content in the Tvärminne archipelago in southern

Finland ( Bernes 2005) than in our study area, which could be stimulating annual algal growth ( Worm & Lotze 2006). P. littoralis appears to be a strong competitor irrespective of wave exposure, since we did not see any differences between the sheltered and exposed sites for this species. This assumption is supported by observations made by Lotze et al. (1999), along with the demonstrated plasticity of this species to different environmental conditions ( Müller & Stache 1989). We did not find any specific correlation between P. littoralis and any of the macrofaunal species, probably because the alga had a similar biomass across both exposures and on all sampling occasions. In early spring, Ulva intestinalis L. has been shown to be superior to P. littoralis in occupying space ( Lotze et al. 2000), and grazing experiments have shown that P. littoralis is preferred by gammarids as a food source over Ulva, Ceramium, Cladophora, Fucus and Furcellaria ( Orav-Kotta et al. 2009). Although contradictory to our results, these findings may still support the results of our study. Among the first faunal species to occur in high numbers was from Hydrobiidae. Being a grazer, it may have indirectly supported the growth of P.

Sequence reagents and all other reagents and chemicals were from Calbiochem-Merck (Darmstadt, Germany). Tetravalent anti-bothropic (B. jararacussu, Bothrops jararaca, find more Bothrops neuwiedi and Bothrops alternatus) and monovalent anti-crotalic (C. d. terrificus) horse antivenom were produced and kindly provided by the Vital Brazil Institute, Niteroi, RJ, Brazil. Two libraries of sixty-nine, 14-mer peptides were designed to represent

a consecutive overlapping coverage that was offset by nine amino acids across the entire coding region (121–122 amino acids) of the three PLA2s present in the venom of B. jararacussu. Sequences were obtained from the UniProtKB – Protein knowledgebase (http://www.uniprot.org/): BthTX-I (Swiss-Prot ID.: Q90249), BthTX-II (Swiss-Prot ID.: P45881) and BthA-I (Swiss-Prot ID.: Q8AXY1). The peptides were automatically prepared onto Amino-PEG500-UC540 cellulose membranes according to standard SPOT synthesis protocols ( Frank, 2002) using an Auto-Spot Robot ASP-222 (Intavis Bioanalytical Instruments AG, Köln, Germany). In brief, coupling reactions were followed by acetylation

with acetic anhydride (4%, v/v) in N, N-dimethylformamide to render peptides unreactive during the subsequent steps. Enzalutamide After acetylation, Fmoc protective groups were removed by the addition of piperidine to render nascent peptides reactive. The remaining amino acids were added by this same process of coupling, blocking and deprotection until the expected desired peptide was generated. After the addition of the last amino acid in the peptide, the amino acid side chains were deprotected

using a solution of dichloromethane–trifluoracetic acid–triisobutylsilane (1:1:0.05, v/v/v) and washed with methanol. Membranes containing the synthetic peptides were either probed immediately or stored at −20 °C until needed. Negative controls [without peptide; IHLVNNESSEVIVHK (Clostridium tetani) precursor peptide] and positive controls were included in each assay. SPOT membranes were washed with Cyclin-dependent kinase 3 TBS (50 mM Tris-buffer saline, pH 7.0) and blocked with TBS-CT (50 mM Tris-buffer saline, 3% casein, 0.1% Tween 20, pH 7.0) at room temperature under agitation or overnight at 4 °C. After extensive washing with TBS-T (50 mM Tris-buffer saline, 0.1% Tween 20, pH 7.0), two membranes presenting the same peptide library were incubated separately for two hours with either horse anti-crotalic or anti-bothropic antivenom (1:250) in TBS-CT and them washed again with TBS-T. Afterward, the membranes were incubated with alkaline phosphatase-labeled sheep anti-horse IgG (1:5000 in TBS-CT) for one hour, and then washed with TBS-T and CBS (50 mM citrate-buffer saline, pH 7.0). Chemiluminenscente CDP-Star® Substrate (0.25 mM) with Nitro-Block-II™ Enhancer (Applied Biosystems, USA) was added to complete the reaction. Chemiluminescent signals were detected on MF-ChemiBis 3.2 (DNR Bio-Imaging Systems, Israel) at a resolution of 5 MP.