The absorbance was measured at 532 nm and results were expressed as MDA equivalents formed by Fe2+ and

H2O2. Nitric oxide was generated from spontaneous decomposition of sodium nitroprusside in 20 mM phosphate buffer (pH 7.4). Once generated NO interacts with oxygen to produce nitrite ions, which were measured by the Griess reaction (Basu and Hazra, 2006). The reaction mixture (1 ml) containing 10 mM sodium nitroprusside (SNP) in phosphate buffer and ATR at different concentrations were incubated at 37 °C for 1 h. A 0.5 ml aliquot was taken and homogenized with 0.5 ml Griess reagent. The absorbance of chromophore was measured at 540 nm. Percent inhibition of nitric oxide generated was measured by comparing the absorbance values of negative controls (only 10 mM sodium nitroprusside and vehicle) learn more and assay preparations. Results were expressed as percentage SB203580 mouse of nitrite formed by ATR alone. The ability

of ATR to scavenge H2O2 (“catalase-like activity” or “CAT-like activity”) was measured as described previously (Aebi, 1984). Briefly, H2O2 diluted in 0.02 M phosphate buffer (pH 7.0) to obtain a 5 mM final concentration was added to microplate wells in which different concentrations was placed. The microplate was immediately placed to monitor the rate of H2O2 decomposition in the microplate reader set at 240 nm. The ability of ATR to scavenge superoxide anion (“superoxide dismutase-like activity” or “SOD-like activity”) was measured as previously described. ATR was mixed to native purified catalase (100 U/ml stock solution) in glycine buffer (50 mM, pH 10.2). Superoxide generation was initiated by addition of adrenaline 2 mM and adrenochrome formation was monitored at 480 nm for 5 min at 32 °C. Superoxide production was determined by monitoring the reaction curves of samples and measured as percentage of the rate of adrenaline auto-oxidation into adrenochrome (Bannister and Calabrese, 1987). SH-SY5Y cells were cultured in 10% FBS DMEM/F12 medium. Cells were used for cytotoxicity measurements when reached 70–90% confluence. Cells were treated with different concentrations

of ATR alone or in the presence of H2O2 400 μM for 3 h, and cell viability mafosfamide was assessed by the MTT assay. This method is based on the ability of viable cells to reduce MTT (3-(4,5-dimethyl)-2,5-diphenyl tetrazolium bromide) and form a blue formazan product. MTT solution (sterile stock solution of 5 mg/ml) was added to the incubation medium in the wells at a final concentration of 0.2 mg/ml. The cells were left for 45 min at 37° C in a humidified 5% CO2 atmosphere. The medium was then removed and plates were shaken with DMSO for 30 min. The optical density of each well was measured at 550 nm (test) and 690 nm (reference). Data are expressed as mean ± SEM. The obtained data was evaluated by one-way analysis of variance (ANOVA) followed by Tukey’s test. All tests were performed in triplicate. Data analyses were performed using the GraphPad Prism 5.

Nestes doentes, o risco clínico de progressão para CHC foi assumido como idêntico ao do doente com HBC e carga viral indetectável. Quarto, em relação ao risco de progressão da doença foi assumido que doentes em supressão viral não estariam em risco de progressão de HBC para CC e de CC para CD9 and 10. Em doentes com carga viral detectável, o risco de progressão assume-se independente do nível de replicação viral. Tal pressuposto não acautela, portanto, o facto de a progressão poder depender, de forma mais gradual, do nível dessa replicação46. Efetivamente, no estudo REVEAL52, a razão de chances (odds ratio) do número de casos de cirrose, face a doentes

com ADN-VHB selleck screening library < 1000 cópias/mL, aumenta proporcionalmente com o nível de ADN-VHB. Acresce que as taxas de progressão utilizadas são também uma aproximação. As taxas de progressão de HBC para CC (de acordo com o padrão de AgHBe) foram assumidas como sendo iguais aos limites superiores reportados por de Francis et al. 31 d. Para as taxas de progressão de CC para CD (assumida como idêntica selleckchem nos 2 padrões de AgHBe) foram utilizados os dados reportados por Idris et al. 3 à semelhança do pressuposto utilizado no estudo de custo-efetividade do tratamento da HBC em Espanha 14. Quinto, em linha com os resultados publicados por Liu et al.53, assume-se que o risco clínico de CHC ou TH é dependente

da carga viral mas independente do padrão de AgHBe. Relativamente à diferenciação do risco clínico de CHC, em função da supressão viral ou ausência desta, deve referir-se que, no estudo recentemente publicado por Papatheodoridis et al.54, não foi encontrada evidência Sunitinib clinical trial de que a supressão

viral prevenisse o CHC em doentes AgHBe-negativos com cirrose. Apesar das limitações e estimativas necessárias para concretizar este estudo, os resultados alcançados demonstram a dominância de uma das terapêuticas avaliadas. Concretamente, de acordo com a análise realizada o medicamento TDF é uma alternativa dominante, quando comparado com ETV no tratamento oral da HBC, uma vez que gera menores custos e maior efetividade. A análise de sensibilidade confirma este resultado para diferentes cenários em que adotamos variações de grande amplitude dos parâmetros do modelo (custos e efetividade). Assim, os resultados ora produzidos constituem um importante contributo como informação de apoio à decisão terapêutica, numa ótica de valorização e otimização dos recursos disponíveis (e escassos) no sistema de saúde português. O estudo de avaliação económica foi financiado pela empresa Gilead Sciences. O estudo foi desenvolvido de forma independente, sem que lhe tivessem sido impostos quaisquer condicionalismos sobre os resultados por parte do financiador. Assim sendo, as opiniões aqui expressas são fruto da análise e interpretação dos autores e não refletem necessariamente outros pontos de vista. J. Areias, A. Carvalho, G. Macedo, R. Marinho, L.

The numerical oscillations visible in the Fluidity output become negligible at 1000 m resolution, with little difference between results at resolutions between 1000 m and 125 m Veliparib ic50 ( Fig. 2). The observed numerical oscillations are caused by the sharpness of the leading and trailing edges of the slide, where minimal smoothing of 1000 m was used ( Haugen et al., 2005). Increasing

the smoothness of these edges (by increasing S in (9)) removes the oscillations. Clearly, the mesh resolution must be high enough to capture the smoothing length or the slide will have an effective flat front. To check that this was the cause of the spurious oscillations, the 5000 m resolution case was re-run with a smoothing length of 7500 m. The results show much reduced oscillations, but with the

wave form shifted due to the new location of maximum height ( Fig. 2). This experiment confirms the correct implementation of the boundary condition and shows how the assumed shape of the slide dictates the mesh resolution required in the slide area. A slide with steeper leading and trailing edges requires higher spatial resolution to eliminate numerical oscillations. To extend our validation 17-AAG in vivo of Fluidity’s new slide-tsunami model to three dimensions, we also replicated a simulation of landslide generated waves that are only weakly dispersive (Ma et al., 2013). Recent work by Ma et al. (2013) simulated the wave train produced by a rigid-block model in a three-dimensional domain on a constant slope. We can therefore compare Fluidity to the results shown in Ma et al. (2013). The domain is 8 ×× 8 km, with a constant slope of 4°. We set the minimum depth to be 12 m and the maximum to be 400 m. We used a horizontal model resolution

was 25 m in x   and y   and explored the influence of vertical resolution by performing simulations with 1–4 layers. Ma et al. (2013) use a different slide geometry to that described above, based on the work of Enet and Grilli (2007). The slide geometry is given by: equation(13) hs=hmax1-∊1coshkbx1coshkwy-∊where kb=2C/b,kw=2C/wkb=2C/b,kw=2C/w and C=acosh(1/∊)C=acosh(1/∊). The slide has length b=686b=686 m, width w=343w=343 m and thickness hmax=24hmax=24 ADP ribosylation factor m. The truncation parameter, ∊∊ is 0.717. The slides moves according to: equation(14) s(t)=s0lncoshtt0where s0=ut2/a0,t0=uta0,a0=0.27 m s−2, and ut=21.09ut=21.09 m s−1 as detailed in Ma et al. (2013). We use these definitions of the slide height and speed for comparisons to Ma et al. (2013). The resulting wave is very similar in magnitude and waveform to that shown in Ma et al. (2013), even using only a single layer in the vertical (Fig. 3). Convergence of the Fluidity model results is observed for three or more element layers (c.f. 40 layers used by Ma et al. (2013)), indicating that the wave is only weakly dispersive. In more detail, Fluidity produces slightly lower amplitude waves than those reported by Ma et al. (2013) (Fig.

There are many precedents for protection of these types of species SP600125 purchase in the terrestrial world; migratory birds are vigorously protected by some countries

while others actively hunt them (e.g. Fox and Madsen, 1997) and terrestrial parks do not protect the entire range of migratory mammals such a wildebeest (e.g. Thirgood et al., 2004). The Convention on the Conservation of Migratory Species of Wild Animals (CMS) is an environmental treaty within the United Nations Environmental Programme that focuses on the conservation and sustainable use of migratory animals and their habitats. CMS is currently engaged in efforts to develop a global conservation instrument for migratory sharks as well as addressing issues facing cetaceans and turtles, including bycatch. The pelagic realm represents the largest global ecosystem and 99% of the Earth biosphere volume (Angel, 1993) and is the least protected marine habitat (Game et al., 2009). It has become increasingly apparent Selleck AZD2281 that the structure and function of this ecosystem has significantly changed largely

due to fishing (Coleman and Williams, 2002, Hyrenbach et al., 2000, Myers and Worm, 2003 and Verity et al., 2002). Based on the greater scientific understanding of the nearshore environment, the most obvious solution to this problem is a no-take MPA. However, pelagic species and habitats are generally thought to be less amenable to spatial protection measures, a view that has translated into a lack of closed area designations within this environment (Day and Roff, 2000 and Game et al., 2009). Two aspects of the pelagic system have fostered the prevailing belief that the application of area closures is an inappropriate management approach; (1) the potentially highly migratory nature of many of the species that inhabit the pelagic system (Boersma and Parrish, 1999) and (2) the ephemeral nature of the physical processes that drive pelagic biological distributions (Etnoyer et al., 2004), though such models fail to adequately consider aspects of habitat heterogeneity and the effects of fishers’ behaviour

(Apostolaki et al., 2002 and Roberts and Sargant, 2002). Habitat heterogeneity is particularly true Adenosine around oceanic islands, with the island mass effect resulting in localised increases in oceanic productivity (e.g. Doty and Oguri, 1956, Hargraves et al., 1970, Gilmartin and Revelante, 1974, Simpson et al., 1982, Le Borgne et al., 1985 and Hernández-León, 1988). There are various theories (reviewed in Genin, 2004) as to why these islands are hotspots of pelagic biodiversity (Worm et al., 2003), particularly for apex predators (Stevenson et al., 2007). Seamounts can perform a similar function (Morato et al., 2008) and have been shown to host populations of bigeye (Holland et al., 1999, Itano and Holland, 2000 and Morato et al., 2008), yellowfin (Holland et al., 1999 and Itano and Holland, 2000) and skipjack tuna (Fonteneau, 1991 and Morato et al., 2008).

, 2010). More recently, Operation Cleansweep (www.opcleansweep.org), a joint initiative of the American Chemistry Council and Society of the Plastics Industry, is aiming for industries to commit to zero pellet loss during their operations. Within the marine environment, plastic is widely considered the primary constituent of ‘marine debris’, a category that includes both anthropogenic litter (e.g. glass, metal, wood), and naturally occurring flotsam (e.g. vegetation, pumice; Barnes et al., 2009, Moore, 2008, Ryan et al., 2009 and Thompson et al., 2004). However, PD-0332991 clinical trial small plastic debris (<0.5 mm

in diameter) is considered a widely under-researched component of marine debris (Doyle et al., 2011) due to the difficulties in assessing the abundance, density and distribution of this contaminant within the marine environment. Quantifying the input of plastics Vemurafenib order into the marine environment is precluded by the array of pathways by which plastics may enter the oceans and would require accurate timescales of the length at which plastics

remain at sea prior to degradation (Ryan et al., 2009). Meanwhile, quantifying debris that has already reached the marine environment is complicated by the vastness of the oceans compared to the size of the plastics being assessed. Spatial and temporal variability owing to oceanic currents and seasonal patterns further complicate this issue (Doyle

et al., 2011 and Ryan et al., 2009). Nevertheless, a suite of sampling techniques has been developed that allow the presence of small plastic debris to be determined. These include: (1) beach combing; (2) sediment sampling; Selleckchem Dolutegravir (3) marine trawls; (4) marine observational surveys; and (5) biological sampling. Beach combing is considered the easiest of the available techniques to conduct, requiring little logistical planning and relatively low costs (MCS, 2010). Typically carried out by researchers and environmental awareness groups, this technique involves collecting and identifying all litter items, in a systematic approach, along a specified stretch of coastline. By repeating the beach combing process on a regular basis, accumulation of plastic debris can be monitored over time (Ryan et al., 2009). This technique is particularly useful for determining the presence of macroplastics and plastic resin pellets, termed ‘Mermaid’s Tears’ by beach combers, but microplastics, especially those too small to be observed by the naked eye, are likely to go unnoticed using such a technique.

Saccades are initiated if and only if the activity of movement neurons reaches a specific and constant threshold activation level independent of the response time 7, 9 and 11]. Fixation neurons are active during fixation and exhibit decreased discharge preceding saccades 12 and 13]. Neurons that participate in controlling movement generation must fulfill two criteria. First, neurons must be active differently when movements are generated or suppressed. Second, the change in activity on canceled trials must occur before SSRT. Some FEF http://www.selleckchem.com/products/DAPT-GSI-IX.html and SC neurons fulfill both of these criteria. On trials where the monkeys are able to respond to the stop signal and

inhibit the saccade, the activity of movement neurons stops increasing and starts to decline before the SSRT elapsed. The likely source of this inhibition is the simultaneous increased activity of fixation cells that also occurs before the SSRT elapses [2]. While our knowledge of response inhibition in the oculomotor system is fairly advanced, we do not understand inhibitory control of skeletomotor movements nearly as well. This is an important unresolved question, because there are a number of significant differences between the oculomotor Ganetespib datasheet and skeletomotor system both in the structure and complexity of their plant and their respective control systems. An important current

research aim has been therefore to investigate the mechanisms of response inhibition of skeletomotor movements. A crucial question is where exactly

in the brain the inhibition of skeletomotor movement preparation takes place and if the mechanism of this inhibition is similar to what is found in the oculomotor system. On multiple levels of the oculomotor system, there are neurons that serve as an inhibitory gate for producing eye movements: in premotor structures (fixation cells in FEF, SC), in the output of the basal ganglia (substantia nigra pars reticulate; SNr), and in the brainstem saccade generator (omnipause neurons) [14]. Functionally similar levels of the skeletomotor Nintedanib (BIBF 1120) system have been recently investigated and different hypothesis regarding inhibitory control mechanisms have been suggested. Pyramidal cells in primary motor cortex (M1) begin to discharge before the EMG burst in agonist muscles and movement onset 15 and 16]. The activation of corticospinal neurons is necessary for initiating and generating skeletomotor movements and stopping such a movement requires fundamentally that the activity in corticospinal neurons is either suppressed or rendered ineffective (Figure 1). M1 and premotor cortex (PMC) seem therefore a likely site of inhibitory control of movement preparation. Application of GABA antagonists to PMC reduced the ability of monkeys to withhold well-trained arm movements to visual targets [17].

These alterations directly increased the rate of biliary sterol excretion and increased

uptake of LDL cholesterol by the liver via the up-regulation of LDL-R. This study was supported by the National Council of Scientific and Technological Development (CNPq, Brazil; CNPq No. 480068/2009-7). We thank Maria Terezinha Bahia (Chagas’ Disease Laboratory, Federal University of Ouro Preto) for the use of the real-time PCR ABI 7300 equipment (Applied Biosystems). M.O.S and M.L.P were sponsored by a fellowship from CAPES and CNPq, respectively. We are also grateful to Rinaldo Cardoso dos Santos for his suggestions and careful review of the manuscript. “
“There has been an error with regard to Fig. 1. The orientation selleck chemicals llc of ICP gene cassette is given from EcoRI to HindIII where it should be from HindIII to EcoRI. This error is deeply regretted. The correct map of T-DNA is given below. “
“Acerola (Malpighia emarginata D.C.), also known as Barbados Cherry, is a tropical

fruit of great economic and nutritional value because of its high content of vitamin C, which is associated with the presence of carotenoids, anthocyanins, iron and calcium. Acerola’s consumption in natura is limited because it is a small fruit with relatively large seeds and is very perishable. The fruit, however, has a good pulp yield, facilitating the development of several click here industrial products. Acerola has been processed in the form of juices, jams, ice creams, syrups, liqueurs and fruit syrups, among other products ( Soares Filho & Oliveira, 2003). In this context, processed products, such as frozen pulp and concentrated pulp, have economic importance; pulp production is a profitable activity that allows the freshly harvested perishable fruit to be stored and reprocessed off-season. The preservation of nutritional constituents during processing represents a major challenge for the traditional

techniques of pulp production. Processing generally involves heat treatment that can reduce the nutritional and organoleptic quality of the product. Over the years, new processing technologies have emerged to reduce or to eliminate the exposure SPTLC1 of the fruit to heat. Ohmic heating is one alternative pulp pasteurization process. This technology can provide rapid and uniform heating, resulting in less thermal damage to labile substances such as vitamins and pigments (Castro et al., 2004 and Sarang et al., 2008). Ohmic heating is defined as a process where electric currents pass through foods to heat them by internally generated energy, without involving any heating medium or heat transfer surface (Castro, Teixeira, Salengke, Sastry, & Vicente, 2003). This heating technology is particularly interesting for viscous products and foods containing particulates because it simultaneously generates heat in both phases and does not need to transfer heat either through a solid–liquid interface or within a solid (de Alwis and Fryer, 1990, Imai et al.

RNA was reverse transcribed with oligo(dT) primers using the SuperScript III First-Strand Synthesis System for reverse transcription–polymerase chain reaction (RT-PCR; Invitrogen) according to manufacturer’s instructions. Primers specific to each gene were designed using the Primer3Plus software (http://www.bioinformatics.nl/cgi-bin/primer3plus/primer3plus.cgi) and synthesized by Invitrogen. The resulting cDNA was amplified by PCR using the gene-specific

primers and the 7300 Real Time PCR System (Applied Biosystems, Foster City, CA) and a QuantiTectTM SYBR Green PCR Kit (Qiagen, Hilden, Germany). A log-linear relationship between the amplification curve and quantity of cDNA in the range of 1 to 1000 copies was observed. The cycle number at the threshold was used as the threshold cycle (Ct). Changes in the expression of mRNA were detected from 2− ΔΔCt using the 7300 Real Time PCR System with Sequence Detection Software (version PLX-4720 mw 1.4; Applied Biosystems). The amount of cDNA in each sample was normalized to the crossing point of the housekeeping gene glyceraldehyde-3-phosphate

dehydrogenase (GAPDH). The following thermal cycling parameters were used: denaturation at 95°C for 10 minutes followed by 45 cycles at 94°C for 15 seconds, 55°C for 30 seconds, and 72°C for 30 seconds. The relative up-regulation of mRNA for each gene in the control was calculated using their respective crossing points with buy Vorinostat the following formula, as previously described [14]: F=2(TH−TG)−(OH−OG)F=2TH−TG−OH−OGwhere

F is the fold difference, T is control, O is the treated cell or tumor, H is the housekeeping gene (GAPDH), G protein-coupled receptor kinase and G is the gene of interest. C-src tyrosine kinase primers: CSK F (forward), 5′-GAATACCTGGAGGGCAACAA-3 Caveolin 3 primers: CAV3 F (forward), 5′-TTTGCCAAGAGGCAGCTACT-3 GAPDH primers: GAPDH F (forward), 5′- GAGTCAACGGATTTGGTCGT-3 To assess the gene expression of caveolin 3 and c-src tyrosine kinase with quantitative RT-PCR, athymic mice harboring U87ΔEGFR brain tumors were killed at 18 days after tumor implantation. The tumor-bearing right hemispheres of the brains were excised and processed for RNA. Student’s t-test was used to test for statistical significance. Data are presented as the means ± standard error. P < .05 was considered statistically significant. All statistical analyses were performed with the use of SPSS statistical software (version 20; SPSS, Inc, Chicago, IL). U87ΔEGFR orthotopic tumors proliferate with aggressive angiogenic growth (Figure 1A). Treatment with bevacizumab reduced angiogenesis in U87ΔEGFR orthotopic tumor tissues in the rat with a regression of tumor size ( Figure 1B). The density of tumor vessels was significantly decreased by bevacizumab ( Figure 1C). The short diameter of tumor vessels tended to be larger, but there was no significant difference ( Figure 1D).

This situation is also likely to be quite different after poisoning with OP nerve agents (e.g. sarin) in which there are no solvents and the onset is much faster, making it likely that AChE inhibition is responsible selleck for all toxic features (Maxwell et al., 2006). Toxicokinetic and dynamic studies indicated that the differences were not due to variation in absorption alone. Red cell AChE activity in pigs poisoned with dimethoate EC40 and dimethoate AI were identical, despite very different poisoning severity. This discrepancy raises questions about the usefulness of this biomarker in OP pesticide poisoning (Eddleston et al., 2009a,

Eddleston et al., 2009b and Eyer et al., 2010). Plasma dimethoate and omethoate concentrations were similar in the first few hours after poisoning with dimethoate EC40, dimethoate AI, and dimethoate AI + cyclohexanone, when differences in toxicity were apparent. The dimethoate and omethoate concentrations after poisoning with dimethoate AI then decreased. The dimethoate concentration after poisoning with the new dimethoate EC formulation

was markedly less than with the other formulations; however, the omethoate concentration was significantly higher and red cell AChE more inhibited, suggesting BIBW2992 manufacturer again that pesticide toxicokinetic differences were not the basis for the differences in toxicity. Plasma cyclohexanol concentrations were substantially lower after poisoning with cyclohexanone alone compared to dimethoate EC40 or dimethoate + cyclohexanone. Plasma cyclohexanone concentrations were also lower after cyclohexanone compared to dimethoate EC40 but less so than its metabolite. These differences

suggest that the presence of dimethoate alters metabolism of the solvent; it is known that dimethoate induces cytochrome P450 activity and its own metabolism (Buratti and Testai, 2007). There was little evidence for dimethoate increasing the absorption of cyclohexanone. The mechanism for the effect of cyclohexanone on dimethoate toxicity is unclear. Both dimethoate AI and cyclohexanone caused a fall in systemic vascular resistance; it is possible that their effects are additive. Alternatively, selleck products the solvent may alter the distribution of the dimethoate and thereby alter toxicity. Further studies are required to address this point. We used arterial lactate concentration as a marker of global toxicity. Its substantial increase in pigs poisoned with dimethoate and cyclohexanone probably represents a combination of tissue hypoxia, hepatic dysfunction reducing lactate clearance, and catecholamine-induced changes in muscle metabolism. The main limitation of this study is that it was performed in anaesthetised minipigs and not humans. An anaesthetised minipig is clearly different to self-poisoned humans and we cannot be sure that the results are a “true reflection” of the human situation.

25, 0.5, 1.0, 2.0, and 4.0 μM) at different phases of the cell cycle based I-BET-762 in vitro on the protocol described by Cavalcanti et al. (2008) with minor modifications. Doxorubicin (0.5 μM) was used as a positive control. All experimental protocols were performed in the presence or absence of colchicine. In the experimental procedures adopted, when PHT was added after 24 h, cells in both G1 and S stages were exposed, while it can be assumed that when PHT was added after 69 h, cells in G2 stage were exposed. When PHT was added in same time of PHA stimulation (in begin of the culture, 0 h) cells were exposed in G1 stage. In order to obtain

a sufficient number of analyzable metaphases, colchicine was added at a final concentration of 0.0016%, 2 h prior to harvesting. The cells were harvested by centrifugation and treated with 0.075 M KCl at 37 °C for 20 min. The cells were then centrifuged and fixed in 1:3 (v/v) acetic acid:methanol. Finally, slides were prepared, air-dried and stained with 3% Giemsa solution (pH 6.8) for 8 min (Moorhead et al., 1960). Slides were analyzed with a light microscope, and structural and numerical CAs were examined in metaphases from the PHT-treated cultures and from the respective controls. The frequency of CAs (in 100 metaphases per culture) and the mitotic index (MI, number of metaphases per 2.000 lymphocytes per culture) MG-132 clinical trial were determined. The differences between experimental groups were compared by one-way analysis of variance

(ANOVA) followed by Tukey’s test. All analyses were performed using the Graphpad program (Intuitive Software for Science, San Diego, CA). The Alamar Blue assay was performed to evaluate the effect of PHT in human lymphocytes. Based on data collected from three independent experiments carried out in duplicate, the IC50 values obtained in human lymphocytes

for PHT and doxorubicin were 5.68 (4.17–7.28) and 1.78 (0.96–3.31) μM, respectively, after 72 h of incubation (Fig. 2). All subsequent experiments were conducted in human lymphocytes at concentrations of 0.25, 0.5, Quisqualic acid 1.0, 2.0, and 4.0 μM. The alkaline comet assay was used to evaluate induction of single-strand and double-strand breaks (DSB) in human lymphocytes. Fig. 3 shows the effect of PHT on the damage index and on damage frequency, as measured by effects on DNA. At 2.0 and 4.0 μM, PHT clearly produced a significant increase in damage index and damage frequency as compared to the control groups. In addition, this increase in damage score occurred in a dose-related manner. CA analysis was performed to evaluate the clastogenic effects of PHT during G1 (Table 1), G1/S transition (Table 2), and G2 (Table 3) of the cell cycle. In addition, the experimental protocols of the CAs were performed in the presence or absence of colchicine to evaluate the action of PHT in the mitotic phase. PHT was clastogenic in all phases of the cell cycle in the presence or absence of colchicine. Chromatid gaps and chromatid breaks were the most frequent CAs.