, 2013, Forenbaher and Miracle, 2006, Greenfield, 2008, Legge and Moore, 2011, Manning et al., 2013, Miracle and Forenbaher, 2006, Özdoğan, 2011, Tringham and Krstić, 1990 and Tringham, 2000). Furthermore, current research suggests that the diffusion of food production was not a simple, straightforward process; different regions underwent distinct histories with varying types of farming

adaptations. In some parts of the Balkans, farming appears as a ‘package’ with a full commitment to plant and animal husbandry as a subsistence system and substantial villages with centuries PD0332991 molecular weight (and in some cases millennia) of occupation (e.g., Bailey, 2000, Legge and Moore, 2011, Marijanović, 2009, Moore et al., 2007 and Perlès, 2001). Other areas display a much greater diversity in both subsistence practices and degree of sedentism, such as in the Iron Gates region, where settled farming communities along the Danube emphasized aquatic resources (Bonsall et al., 2008), or parts of Romania where semi-sedentary pastoral gatherers interacted with more sedentary farmers (Greenfield and Jongsma, 2008), and possibly with indigenous hunter-gatherer groups (Bailey, 2000, Borić and Price, 2013 and Tringham, 2000). The connections between these regions and the

variations in the mechanisms are Selleckchem AUY 922 still a matter of debate. Cultural affinities based on ceramic styles point to the Balkans as a departure point for farming traditions throughout Europe, with interior trajectories exemplified by people who produced

Starčevo pottery toward central Europe, and Mediterranean linkages in the form of Impresso wares (pottery decorated with shell and non-shell impressions) throughout the Adriatic and into the Western Mediterranean ( Rowley-Conwy, MRIP 2011; see also Manning et al., 2013). In this way, the Balkan Peninsula is an ideal area to examine the varied effects of agricultural production on landscapes, human and animal populations, and issues of degradation. This diversity, however, also poses some key challenges in identifying regional trends within the forest of specific or local historicity. In all cases, early farming villages in the Balkans share some basic features of sedentary life and reliance on domesticated plants and animals for subsistence. Specifics in the relative proportions of domestic species in bone assemblages from these sites, the contribution of wild species to diets, and the interplay between species reflect not only variations in cultural adaptations but also ecological dynamics in interior and coastal regions. Table 1 and Fig. 2 summarize the available published data on the relative proportions of wild and domestic animals at a number of Early Neolithic villages in the region.

The animals were maintained under a controlled temperature (22 ± 2 °C) and with free access to water and commercial feed. The animals were kept in the experimental room for at least 1 week prior to testing for adaptation. The experiments were performed DAPT supplier in accordance with the guidelines established for the care of laboratory animals. This study was approved by the “Research Ethics

Committee on Animal Use” at the Federal University of Rio Grande do Norte, under protocol no. 003/2012. Chitosan (85% deacetylated, molecular weight: 90–190 kDa), aluminum hydroxide, TPP and T. serrulatus venom were purchased from Sigma-Aldrich Co. (St. Louis, Mo.) BCA Protein Assay Kit was purchased from Pierce Biotechnology (Woburn, MA) and Mouse IgG total ELISA Kit from eBioscience (San Diego, CA, USA). All other reagents and solvents used were of analytical grade. The electrophoretic profile of T. serrulatus venom was determined by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE), using the minigel system

(Mini-ProteanTM II) ( Laemmli, 1970). Dasatinib research buy The relative molecular mass of proteins was determined with polyacrylamide gel by comparing the electrophoretic migration pattern of a protein mixture obtained commercially (Gibco-BRL Life Technologies, Gaithersburg, MD, USA). The gels were stained in a solution of “”Commassie Brilliant Blue”" R-250, for 45 min and washed in bleach solution until the disappearance of background staining and then scanned (Morrissey, 1981). The cross-linked chitosan nanoparticles for the incorporation of T. serrulatus venom were obtained through the method of ionic gelation. Thus, a 0.1% w/v tripolyphosphate (TPP) in water obtained by reverse osmosis (<1.3 μS cm−1) was dripped in a 0.1% w/v of chitosan in a 0.175% w/v acetic acid solution under

magnetic stirring. When a spontaneously formed opalescent suspension was obtained, this remained under magnetic stirring at room temperature Glutamate dehydrogenase for 30 min. The particle size and zeta potential were determined using zeta sizer equipment (ZetaPlus – Brookhaven Instruments Corporation, EUA). A polydispersity smaller than 0.5 was required for all the equipment. Triplicate samples were analyzed and the arithmetic mean value of the three was adopted. Chitosan nanoparticles separated from suspension were dried in a freeze dryer, their FT-IR were taken with KBr pellets on Perkin Elmer Spectrum one FT-IR. For protein loading in chitosan nanoparticles, T. serrulatus venom in different ratios (5 and 10%) relative to used chitosan concentration, were dissolved previously in the TPP solution, which was maintained at a temperature of 20 ± 2 °C ( Gan and Wang, 2007) before the nanoparticle preparation procedure (Section 2.3.2). The different T.

Other toxins acting on Sodium channel site III, as Tx2-6, fail to induce priapism possibly by pharmacokinetic reasons but this should still be investigated experimentally. The question whether Metformin price these other toxins that act on Sodium channel site III interfere with NO/NOS/cGMP system was never addressed and could eventually explain why these other toxins don’t induce priapism. The cascade of events triggered by the toxin is currently under investigation

in our laboratory. It is clear though, that more investigations are needed to identify the ultimate mechanism of action involved in the erectogenic effect as well as the local consequences of a long-term use of this toxin. We conclude that crude venom and pure Tx2-6 toxin seem to produce identical effects on the organs examined suggesting that the possible cause of death is lung intra alveolar hemorrhage; toxin and crude venom seem to exert mild to moderate effects on brain tissue as suggested by our previous results (Troncone et al., 2011). In addition, the observed edema could be alternatively

attributed to the respiratory impairment caused by the severe lung hemorrhage. We gratefully acknowledge Dr. Daniel Pimenta and Dr. Isabel F. C. Correia (Biochemistry Laboratory – I. Butantan) for mass spectrometry of fractions and amino acid sequencing and the technical support of Mr. Wilson B. D’Ávila. Supported by research grants from FAPESP No. 98/02039-0 Crizotinib cell line to LRPT and 06/57922-3 to MS. “
“The PTK6 skin of fish constitutes a pivotal immunological protection against the external environment. The layer of mucus on the fish surface, considered the first line of defence, participates in a number of functions including disease resistance, respiration, ionic and osmotic regulation, locomotion, reproduction, communication, feeding

and nest building (Negus, 1963, Ingram, 1980, Shephard, 1994 and Zhao et al., 2008). The mucus, such as that produced by the skin of the stingrays, has a complex set of components, which may include amino acid residues, peptides, complex carbohydrates, glycopeptides, glycolipids and other chemicals (Klesius et al., 2008, Alexander and Ingram, 1992 and Birkemo et al., 2003). Fish epidermal mucus was found to display antimicrobial activity against broad range of infectious pathogens (Mozumder, 2005 and Hellio et al., 2002). We recently described the antimicrobial activity of catfish Cathorops spixii mucus ( Ramos et al., 2012). Moreover, histone H2B and two ribosomal proteins are examples of proteins with antimicrobial activity that have been isolated from epidermal mucus of Atlantic cod ( Bergsson et al., 2005). Members of some families of antimicrobial peptides (AMPs) were also found to be important innate defence components in the epidermal mucosal layer of Moses sole fish (Pardachirus marmoratus) ( Oren and Shai, 1996), winter flounder (Pleuronectes americanus) ( Cole et al.

39. Biopsies of all suspicious lesions are recommended to exclude dysplasia. This 35-year-old man with an indeterminate colitis had a 1-cm inflammatory-appearing polypoid lesion within a colitic area. Biopsies excluded dysplasia and confirmed chronic inflammation. Figure options Download full-size image Download high-quality image (189 K) Download as PowerPoint slide Fig. 40. Inflammatory polyps. In addition to enhancing the

border, chromoendoscopy makes it easier to examine the mucosal surface of lesions and facilitates the recognition of inflammatory patterns. Below, a few examples of hyperplastic polyps and sessile serrated adenomas/ polyps are presented. Figure options Download full-size image Download high-quality image (211 K) Download as PowerPoint slide Fig. 41. Hyperplastic polyp. Figure

options Download full-size image Download high-quality image (275 K) Download as PowerPoint slide Fig. 42. Sessile serrated adenoma/polyp. Figure IDO inhibitor www.selleckchem.com/products/Rapamycin.html options Download full-size image Download high-quality image (466 K) Download as PowerPoint slide Fig. 43. Sessile serrated adenoma/polyp. Figure options Download full-size image Download high-quality image (471 K) Download as PowerPoint slide Fig. 44. Depressed neoplasm. Visualization of the depressed morphology required the application of chromoendoscopy. The depressed center of this nonpolypoid (0-IIc) lesion with LGD can only be shown by spraying indigo carmine to show it Demeclocycline pooling in the depressed part. Figure options Download

full-size image Download high-quality image (278 K) Download as PowerPoint slide Fig. 45. See above (Fig. 44). Visualization of the depressed morphology required the application of chromoendoscopy. It is important to understand that the depressed area likely contains the most advanced histology. Thus, both biopsy can be targeted and removal can be optimized. Figure options Download full-size image Download high-quality image (343 K) Download as PowerPoint slide Fig. 46. (A–D) Polypoid neoplasms can be endoscopically resected. Whenever possible, lesions less than 2 cm in size should be resected in one piece (ie, en bloc) using EMR. The use of chromoendoscopy can facilitate delineation of the neoplastic borders and ensure complete resection. Following resection, the mucosa around the site should be biopsied to exclude the presence of invisible dysplasia. Figure options Download full-size image Download high-quality image (248 K) Download as PowerPoint slide Fig. 47. Dynamic injection can be useful in IBD. Sessile and non-polypoid colorectal lesions in patients with IBD may be best cut after injection. Using the dynamic injection technique the injection is directed into the lumen, to mold the fluid bleb formation. Using slight upward tip deflection, the lumen is suctioned and the needle catheter nominally pulled back while directing the injection into the lumen. In this case, the lesion lifted nicely to form a large bleb.

[21] and [22]. The stover sugar hydrolysate contained various impurities, including fine solid particles, degradation compounds (acetic acid, furfural, 5-hydromethylfurfural, phenol derivatives etc.), sodium sulfate salt from neutralization of sulfuric acid, and cellulase

enzyme residues. These impurities would significantly reduce the activity and life time of nickel catalyst in the consequent hydrogenolysis of sugars into polyols [23] and [24], unless an extensive purification step was processed. Similar purification procedures used for the corn-based glucose preparation were applied to the stover Staurosporine nmr sugar hydrolysate, including the two major steps: decolorization and desalting. In the first purification step, the hydrolysate was adsorbed by activated charcoal to remove the pigmented impurities which gave the hydrolysate dark black color. Addition of activated charcoal at 3% (w/w) dosage was found to be sufficient to remove the pigmented impurities. Table

1 shows that all furfural and most 5-hydroxymethylfurfural were removed from the hydrolysate, while the sugars and organic acids maintained the same or even increased slightly due to the water loss. The results were in agreement with the previous studies [25] and [26]. It is worth noting that the protein content in the hydrolysate selleck compound was not detected after decolorization, indicating that the cellulase enzyme protein in the hydrolysate was completely removed by the activated charcoal. In the second purification

step, the Na2SO4 and other salts in the decolorized stover sugar hydrolysate were removed by ion exchange absorption in two steps: the positive ions such as Na+ were removed by the Protein tyrosine phosphatase cation resins 732, and then the negative ions such as SO42− were removed by anion resins D315, respectively. Fig. 3(a) shows that the conductivity of the hydrolysate elute increased quickly in the first 2 min of cation ions exchange, indicating the exchanging of positive ions in the hydrolysate with hydrogen ions on resins started. The hydrolysate conductivity was maintained at a higher value (44,000 μS/cm) until the resins were saturated by the ions such as Na+. Then the hydrolysate was sent for anion ion exchange using the resin D315 to remove negative ions such as SO42−. Fig. 3(b) shows that the conductivity of the stover sugar hydrolysate decreased sharply from 44,000 μS/cm to 4000 μS/cm, indicating the negative ions such as SO42− were sufficiently absorbed by D315 resins. No apparent change of the sugar concentrations (glucose and xylose) between the purified and the original hydrolysates, implying that the sugar loss was negligible during the purification steps. The catalytic hydrogenolysis of stover sugars for short-chain polyols synthesis was conducted as shown in Table 2. The polyols product here refers to ethanediol, 1,2-propanediol, and butanediol.

Nuclear Magnetic Resonance spectroscopy, with the notorious limitation in size due to relaxation-dependent line broadening, seems unsuitable for these particles. Yet, the progresses in both hardware and methodology seen in the last two decades suggest that the goal of studying high-molecular-weight RNP complexes by NMR might be in reach. Fulvestrant chemical structure The first requirement for applying NMR to large particles is the ability to observe the NMR resonances of their components. For the protein parts of the RNP complexes this task is no longer a challenge. The development of the methyl TROSY (transverse relaxation-optimized spectroscopy) technique [18] has

made the observation of methyl groups of Ile, Val, Leu and Met residues feasible Selleckchem Epigenetic inhibitor in molecules as large as 670 kDa [19]. The motion of methyl groups is partially decoupled

from the slow overall tumbling of the complex (low order parameter S2methyls); in addition, like in TROSY spectroscopy, a simple HMQC (heteronuclear multiple quantum correlation) experiment achieves transfer of magnetization among slowly relaxing coherences in the CH3 spin system [18]. Both these facts, together with the steadily improving sensitivity of the instrumentation through the development of high field magnets (a 1.2 GHz magnet is expected to be commercialized in 2016) and of better probe heads, allowed detection of methyl groups in high-molecular weight protein complexes at concentrations as low as tens of micromolar. In seminal work on the 20S proteasome, the group of L.E. Kay has acetylcholine demonstrated that methyl group resonances can be used to probe intermolecular interaction interfaces at atomic precision [19]. This technique requires selectively

13C, 1H labeling of side-chains methyl groups in an otherwise fully deuterated protein. Using commercially available precursors it is possible to obtain 13C, 1H labeling of one of the two prochiral methyls of Val/Leu and of Ile-δ1[20]. Additional strategies have been developed to obtain proS or proR specifically 13CH3 methyl labelled Leu and Val [21] as well as 13CH3 methyl labeling of Ala [22], Met [23] and Ile (γ2) [24]. The assignment of the methyl groups to single amino acid position can either be transferred from single protein domains or sub-complexes, where the classical three-dimensional experiments to correlate side-chains resonances with backbone resonances are still feasible [25], or obtained by single-point mutagenesis. For the RNA part of the RNP complex, the situation is more complex as nucleic acids do not display any moiety with very low order parameters, such as side-chain methyl groups in proteins. On the other hand, the proton density in RNA is not uniform with the base protons of purine being quite isolated in the aromatic ring.

This assumption also yields marginally conservative values of the deficit-volume which is a desirable feature in the design of water resources systems towards ameliorating the drought conditions. It is worthy to mention that Millan and Yevjevich (1971) developed the regression equations for predicting E(LT) and E(MT) which were also tested for the annual and monthly hydrological droughts using Canadian

river flows. These relationships were found reasonably check details reliable although at times they tended to under predict in the range of 3–10%. As a note in the context of analysis of monthly droughts, it is prudent to mention that the values of ρ1 in the SHI sequences were low suggesting a weak dependence structure. Therefore, the first order Markov chain model (Markov chain-1, Eq. (8)) was tried to estimate E(LT). It was noted that the predictions of LT tended to be almost the same as predicted by the extreme number theorem. However, at times the predictions by the extreme

number theorem tended to be marginally higher than the Markov chain-1 model and also be nearer to the observed counterparts. This observation vindicates the applicability of the extreme number theorem on monthly as well as annual basis. In fact as the name reads “theorem of extremes of random numbers of random variables” essentially is meant for random sequences, which is evidenced by the results in the present case (annual flows). It has the capability to perform reasonably well in the presence of weak dependence structure and for this reason Staurosporine mouse it performed satisfactorily even in monthly streamflow series. It was also observed that when the degree of the first order dependence is remarkable (i.e. ρ1 being above 0.5) then the extreme number theorem breaks down and recourse to the Markov chain models,

among others becomes a necessity. The weekly SHI sequences of rivers with negligible lake effects such as those in Atlantic Canada tended to follow AR-1 process, therefore the extreme number theorem based relationships (Eqs. (1), (2), (3), (4) and (5)) were attempted to model E(LT). In general, such a model resulted in consistent under prediction. As noted earlier, the weekly SHI sequences Rapamycin purchase of rivers riddled with significant lake storages tend to obey AR-2 process or even higher order dependence processes ( Table 2). For such rivers, the extreme number theorem does not hold because of a lack of accountability for the second order dependence. Therefore, a second order Markov chain model (Eq. (7)) was envisaged in which the parameters were computed using the counting method ( Sen, 1990 and Sharma and Panu, 2010). The best estimates of the first order probabilities were obtained using the non-standardized weekly flow series ( Table 2).

Opis badania powinien obejmować: wielkość (długość), echostrukturę i echogeniczność nerek, ewentualne poszerzenie układu kielichowo-miedniczkowego (miedniczka i kielichy), szerokość moczowodów oraz wielkość i grubość ścian pęcherza moczowego. Poród dziecka, u którego podejrzewa się poważną wadę wrodzoną układu moczowego, powinien odbywać się w ośrodku referencyjnym III stopnia, zapewniającym możliwość konsultacji urologa i nefrologa dziecięcego. Zaleca

się, by wszystkie dzieci z podejrzeniem prenatalnym wady układu moczowego miały wykonane HSP inhibition badanie ultrasonograficzne jamy brzusznej w pierwszych dobach życia (doba 1.–7.). O terminie badania decyduje stan dziecka i rodzaj podejrzewanej wady (badanie pilne w 1.–2. dobie, a badanie planowe w 3.–7. dobie). Do ustalenia postępowania zalecane jest kolejne badanie ultrasonograficzne jamy brzusznej, które powinno zostać wykonane w terminie 4.–6. tygodni od pierwszego. Do ustalenia właściwego postępowania z noworodkiem niezbędna jest możliwość analizy: ilości wód płodowych, prenatalnej wielkości nerek i szerokości dróg moczowych, stanu klinicznego noworodka (skala Apgar) i wielkości diurezy po porodzie. Poród dziecka

z podejrzeniem poważnej wady wrodzonej układu moczowego (skąpowodzie, brak miąższu obu nerek, zastawki cewki tylnej) powinien odbywać się w ośrodku BIBW2992 molecular weight referencyjnym zapewniającym intensywną opiekę okołoporodową. Należy wykonać badanie USG w pierwszej dobie życia, monitorować diurezę poprzez założenie cewnika do pęcherza moczowego, włączyć profilaktykę zakażeń układu moczowego oraz ocenić czynność nerek poprzez pomiar diurezy godzinowej, a także pomiar stężenia mocznika i kreatyniny w surowicy (z uwzględnieniem wartości tych wskaźników u matki). Konsultacja urologa i nefrologa powinna odbyć Farnesyltransferase się w trybie pilnym (Ryc. 1). Planowa diagnostyka u noworodka w dobrym stanie ogólnym obejmuje badanie ultrasonograficzne w 3.–7. dobie po urodzeniu, co pozwala uniknąć

wyników fałszywie ujemnych spowodowanych przejściowym, fizjologicznym, gorszym nawodnieniem dziecka w 1.–2. dobie życia 2., 3., 4. and 5.. Jeśli w prenatalnym badaniu USG rozpoznano izolowane jedno-lub obustronne poszerzenie układu kielichowo-miedniczkowego (UKM), nie istnieje podejrzenie obecności wady złożonej. Poród dziecka i wstępna postnatalna weryfikacja wady mogą być przeprowadzone w szpitalu rejonowym. Za istotne poszerzenie UKM, wymagające monitorowania, uznaje się poszerzenie miedniczki nerkowej w projekcji A-P powyżej 5 mm w 3.–7. dobie życia i 10 mm w 4.–6. tygodniu lub później. W przypadku izolowanego, niepowikłanego, jednolub obustronnego poszerzenia UKM nie ma wskazań do wykonania cystografii mikcyjnej. Przyczyną poszerzenia UKM u płodu jest najczęściej przeszkoda zlokalizowana na wysokości połączenia miedniczkowo-moczowodowego.

Depending on the location of the sea level pressure centres and the

resulting main flow directions to central Europe, the types ‘North’, ‘South’ and ‘East’ can be distinguished. In addition, all troughs with a north to south axis are classified as meridional circulations. The major types ‘North-East’ and ‘South-East’ are also included in the meridional circulation group because they normally coincide with blocking highs over Northern and Eastern Europe. The meridional circulation group during winter is due to 25% of the Selleck Selumetinib satellite data (Krüger & Graßl 2002) for JFND8589 and 35% for JFND9699, and during summer 38% for MJJA8589 and 39% for MJJA9699. The analysis confirms the same tendencies of cloud albedo changes independently of the circulation group. The changes are in line with the results presented in Krüger & Graßl (2002) and Krüger et al. (2004). The cloud albedo for the zonal and meridional circulation groups during winter (JF and ND) is shown in Figure 1. The tendencies for the zonal as well as the meridional circulation groups appear to be conspicuously connected to PM emission changes on the one hand (during ND) and SO2 emission

changes on the other (during JF). Firstly, a decrease in reflectance from the GSK-3 beta pathway early 1980s to the late 1990s occurs during early winter (ND). The albedo decreases primarily following the reduction in PM emissions in Germany. It is more pronounced for the meridional circulation. The highest cloud albedo in ND during the early 1980s pollution episode can be explained by the existence of the radius effect (Twomey 1974). Enhanced turbulence during ND, as compared to JF, may well have favoured the effective lifting of primary aerosols to cloud level. Nitroxoline The cloud albedo during ND8184 as compared to ND9699 was 4% higher for zonal circulation and even 6% higher for meridional circulation (see Figure 2). Secondly,

the magnitude of the cloud albedo in JF for both circulation groups tends to follow the level of SO2 emissions, which originated mainly from large power plants in the former GDR (as described by Krüger et al. 2004). The highest value of the albedo is for JF8589, which points to the major influence of secondary aerosols. As before, the changes for the meridional circulation group are stronger. The most likely explanation is that the episodes in late winter with the more often stably stratified atmospheres favour the formation of sulphate layers, i.e. haze, which in turn enhance the cloud albedo through the radius effect (Krüger et al. 2004). Thirdly, the trend in cloud reflectance variability seems to be highly influenced by the PM emissions, because of the higher BC content, which can lead to greater absorption and a lower cloud albedo. In addition, secondary particles are contributing to the overall variability through an albedo increase (Figure 2).

, 2009 and Durban et al., 2011; Junqueira-de-Azevedo and Ho, 2002, Kashima et al., 2004, Wagstaff and Harrison, 2006 and Zhang et al., 2006).

More recent application of next-generation sequencing technology (Chatrath et al., 2011, Jiang et al., 2011 and Rodrigues et al., 2012) to transcriptomics will further accelerate this process, as will the increasing ability to directly access the genome through extended read length and targeted sequencing (Glenn, 2011). However, current methods for studying pharmacological activity are generally labour-intensive and the functional characterisation of these new toxins is unlikely to keep pace (not unique to toxins, as the majority of protein sequences in databases lack functional annotation). Computer-generated annotations have been shown to be highly inaccurate (Schnoes et al., 2009) mainly as a result Selleck CT99021 of over-prediction (i.e., annotation to functions that are more specific than the available evidence supports, sometimes naively based on homology to primary structures). This is likely to be the case for most animal toxins, which often retain the ancestral non-toxic structural scaffold, while evolving diverse potent and highly specific toxic activities. In some cases, the substitution of a single amino acid is enough to change the selectivity

for another target (Ohno et al., 1998). In the case of PLA2 toxins, buy 3-MA the ancestral phospholipase activity may be readily predicted while failing to predict the main biological activity of Baricitinib the protein in question. Thus, predicting the function of snake venom proteins based on a common scaffold presents a challenge to bioinformaticians interested

in the analysis of protein sequence–function relationships in general. Solving this problem will have a number of beneficial outcomes as many of the activities of these proteins are of great utility as research tools and potential drugs (Koh et al., 2006), especially in neurological (Sun et al., 2004), anti-cancer (Bazaa et al., 2010 and Lomonte et al., 2010), anti-viral (Fenard et al., 1999 and Meenakshisundaram et al., 2009) and anti-inflammatory (Coulthard et al., 2011) research. In this paper, we report a model-based analysis of the largest dataset of PLA2 Group II toxins to date, comprising 251 protein sequences. Of these, 73 are novel sequences derived from a genome-based survey of PLA2 genes in pitvipers (Viperidae: Crotalinae), including 16 species for which no PLA2 sequences exist in databases. Most of the newly investigated species belong to the Asian Trimeresurus radiation ( Malhotra and Thorpe, 2004), which have been relatively understudied by toxinologists ( Gowda et al., 2006, Soogarun et al., 2008, Tan and Tan, 1989, Tan et al., 1989 and Wang et al., 2005). We used two methods with different conceptual bases.