Spray-dried, water-extracted GJG powder was obtained from Tsumura & Co. (Tokyo, Japan). GJG was approved in 1986 as a drug for clinical use by the Japanese

Ministry of Health, Labour and Welfare. It is produced at the Shizuoka plant which meets Japanese pharmaceutical GMP (good manufacturing practice). The local pharmaceutical administration of Shizuoka Prefecture assesses the GMP status of the plant every 5 years. The plant has had permission for pharmaceutical production for more than 30 years, and the production process has been well validated. Since active substances are still ambiguous, quality control is conducted by quantitation of major components. In the case of GJG, paeoniflorin (moutan bark), loganin (Rehmannia root), and total alkaloids (processed aconite root) are chosen as marker compounds for quality control. Paeoniflorin, loganin, and total alkaloids in 1 g of GJG extract powder used in our experiments were 2.11, c-Met inhibitor Selleckchem GSK3 inhibitor 1.58, and 0.11 mg, respectively. In 10 lots (a total of 20 lots) produced before and behind this lot, paeoniflorin, loganin, and total alkaloids were within ± 10% of the range of this content, and quality was managed satisfactorily. Other physicochemical properties, e.g. loss on drying, water content, ash, heavy metals, etc., were also examined in all lots.

GJG extract is listed in the Japanese Pharmacopeia, and the material used in this study met that description. The general manufacturing procedure of GJG extract powder is as follows. Ten kinds of botanical raw materials are crushed and then weighed in accordance with the mixing ratio as shown in Table S1. The mixture of botanical raw materials is extracted 12 times with ion-exchanged water for 60 min at 100 °C. The extract is centrifuged to obtain a supernatant, which is then concentrated in vacuo. The

concentrated extract solution is dried by a spray dryer. The standard yield of extract powder is around 16% of the total weight of botanical raw materials. A three-dimensional high-performance liquid chromatography (HPLC) profile of a methanol solution of GJG was performed according to our aminophylline previous procedure ( Hattori et al. 2010) and is shown in Fig. 1. 3D-HPLC analysis and LC/MS analysis of the crude drugs involved in GJG are shown in Figs. S1–S3. Seven-week-old male SAMP8 mice were purchased from SLC, Inc. (Shizuoka, Japan) and divided into 2 groups: those fed a normal diet (powdered mouse food; Oriental Yeast Co. Ltd. (Tokyo, Japan; P8 + N group; n = 10)); and those fed a normal diet supplemented with 4% (w/w) GJG (P8 + GJG group; n = 10). As controls, 7-week-old male SAMR1 mice were purchased from SLC and also divided into 2 groups: those fed a normal diet (R + N group; n = 10) and those fed a normal diet supplemented with 4% (w/w) GJG (R + GJG group; n = 11). General conditions and body weight were recorded for all mice.

Twenty-nine items were deleted and nine items were added in total, leaving 62 items to enter psychometric testing. Emphasis was placed upon retaining a sufficient number of items to represent each of the five themes identified. Following expert and patient refinement, two independent item pools were confirmed as suitable to enter psychometric testing. The first item pool contained 23 items asking respondents about their general attitudes toward health websites whilst the second item pool contained 39 items asking the respondent about their attitudes

Epigenetics Compound Library purchase toward a specific health website. All items have a five point response scale (Strongly disagree–Strongly agree). Establishing a robust evidence base for the use of health websites is becoming increasingly important given that patients routinely turn to the web for information and support. This research developed items which will inform a new measure to evaluate the health related effects of websites and create a standardized method to compare health websites. Items constructed were

checked Kinase Inhibitor Library for their applicability across long term conditions, health behaviors and carers and for websites featuring facts and figures, health experiences and discussion forums. This paper documents the steps taken to inform items that may be included in the e-Health Impact Questionnaire. A recent literature review [14] relating to the potential effects of seeing and sharing experiences online and a secondary data analysis of interviews relating Pomalidomide mw to experiences of health were used to generate a range of items. Five themes were identified as relevant to the impact of using health websites containing scientific

information and to websites containing experiential information: (1) Information, (2) feeling supported, (3) relationships with others, (4) experiencing health services, and (5) affecting behavior. Confirmatory data sources were used to triangulate the findings. Comparing themes to issues raised in the focus group transcripts and user panel forms provided more depth in relation to negative aspects of using the internet, for example, becoming isolated from society through the overuse of discussion forums or misdiagnosing symptoms. Using a range of sources to identify and confirm themes provided strong evidence for their inclusion in the item pool. After a period of item selection, the item pool was evaluated by experts in the area of e-health.

The cells retrieved from the surface and from the gel were analysed for their content of lymphocyte subsets by flow cytometry (see below). Freshly isolated, non-adherent or the various migrated cells were labelled with a combination of the following antibodies: anti-CD4-PE, anti-CD8-FITC, anti-CD3-PerCP; anti-CD62L-FITC, CD45RA-PE (all from Becton Dickinson, Oxford, UK), anti-CD45RA-CY5 (Serotech, Oxford, UK), anti-CD4-efluor405; anti-CD8a-efluor605 and anti-CD19-PE-Cy7 (all from eBiosciences, Hatfield, UK)

for 30 min on ice. Labelled cells were spiked with a known volume of Flow-Count Fluorospheres (Beckman Coulter, High Wycombe, UK). Cells were counted and their fluorescence analysed using a Cyan flow cytometer and Summit software (both from Dako). In some cases, cells were enumerated by passing buy Ribociclib the entire sample through the flow cytometer. In this way, find more we could separately count and calculate the percentages of the following subsets that adhered, transmigrated or penetrated into the gels: CD4+ or CD8+ T-cells (CD3+), which were of naive (CD45RA +, CD62L +), effector memory (CD45RA −, CD62L −) or central memory (CD45RA −, CD62L +) phenotypes; CD19+ B-cells (Supplemental Fig. 1). Endothelial cells were incubated with non-conjugated

antibodies against E-selectin (1.2B6) or VCAM-1 (1.4C3; both Dako, Ely, UK) for 30 min at 4 °C, washed and incubated with goat anti-mouse FITC-conjugated secondary antibody (Dako) for 30 min

at 4 °C as previously described (McGettrick et al., 2009b and McGettrick et al., 2010). Fibroblasts were incubated with APC-conjugated anti-ICAM-1 (BD Pharmingen, UK) for 20 min at 4 °C. Subsequently, cells were washed and incubated with enzyme-free cell dissociation buffer (Gibco) for 30 min. The dissociated cells were analysed by flow cytometry and C1GALT1 data were expressed as median fluorescent intensity (MFI). Endothelial mRNA was isolated using the RNeasy Mini Kit (Qiagen, Crawley, UK). Gene expression of the chemokines CXCL9, -10, and -11 was analysed by reverse transcription (RT) PCR, followed by densitometry of product bands run on agarose gel containing ethidium bromide, as described (McGettrick et al., 2009b and McGettrick et al., 2010). Data were expressed as a percentage of the β-actin bands. Variation between multiple treatments was evaluated using analysis of variance (ANOVA), followed by comparison of treatments by Bonferroni (inter-treatment) or Dunnett (comparison to control) test as appropriate. Effects of single treatments were analysed by paired or unpaired t-test as appropriate. P < 0.05 was considered as statistically significant. The level of adhesion to EC was slightly higher for cytokine treated than unstimulated cultures, and co-culture with fibroblasts tended to increase this level, but neither effect was statistically significant (Fig. 2A).

They found that a simple model consisting of age and prior fractures performed as well as FRAX® and the Garvan calculator when BMD was unknown. As in our study, they based assessment on self-reported clinical risk factors; however, they used self-reported incident fractures during 2 years of follow-up while we collected fracture data from national registers. We invited participants from

a random selection in the general population and had a high responder rate (84%). In contrast, the GLOW study group acknowledged that their sample was prone to bias due to the selection of physicians and due to the sampling and recruitment of patients [36]. Also, their model

(with age and prior fracture) was not validated in independent populations. Several other studies have also compared PD0325901 price FRAX® with other more elaborate tools such as the QFracture algorithm [34] and the Garvan calculator [33] and [37] Selleck SCH900776 arriving to the same conclusions as the studies mentioned above. In our study, agreement between the tools with regard to categorizing women into quartiles of risk for major osteoporotic fracture was moderate. However, agreement between the tools in identifying women at the highest quartile of risk for major osteoporotic fracture was high. Approximately 80% of the women classified in the highest risk quartiles by FRAX® were also categorized as highest risk by all the other tools. Sambrook et al. [36] came to a similar conclusion in the GLOW study and our research supports that if women were selected for treatment based on being in the highest quartile of risk, virtually the same women would meet the threshold for treatment regardless of the tool Selleck Enzalutamide used. FRAX® is the most complex tool in this study and incorporate 11 risk factors in the algorithm (and may in addition include BMD), whereas the simpler tools only incorporated

between 2 and 6 risk factors (Table 1). All the tools included age and BMI. Additional variables did not appear to improve the performance of the tools. Both age and BMI are associated with fracture risk, however, age is the strongest risk factor [1]. Our study also showed that even age alone performed as well as the FRAX® tool without BMD. Kanis et al. [23] recently discussed potential pitfalls in external validation of FRAX®. Several studies [33], [35], [38], [39] and [40] compared the AUC of ROC curves across studies. In the present study we compared the AUC of the different predefined tools within the same well defined study population.

Para medir a perceção do estado de saúde e da qualidade de vida usou-se a versão portuguesa da escala de medição Short-Form 36 (SF-36), devidamente validada para a população portuguesa 26, 27, 28 and 29. A recolha de dados efetuou-se entre os meses de abril a julho de 2011. Após o estudo piloto, o questionário foi disponibilizado online e iniciou-se a sua divulgação através dos contactos de correio eletrónico dos investigadores, esperando-se um efeito «bola de neve». A APC também contribuiu ativamente para o estudo, divulgando o questionário, por e-mail, pelos seus associados. De forma a chegar-se GSK1120212 ic50 ao maior número de doentes celíacos

possível, solicitou-se igualmente à Associação Portuguesa dos Nutricionistas a divulgação do questionário pelos parceiros e associados. Alguns blogues dedicados à DC e ao seu Neratinib cost tratamento iniciaram a divulgação do mesmo de forma voluntária. As redes sociais constituíram ferramentas essenciais de divulgação do questionário, que foi partilhado através das páginas pessoais dos investigadores e dos seus contactos e também pela página da APC. Além disso, o questionário incluía um botão de partilha automática pelo Facebook®. Na apresentação do questionário mencionava-se, de forma explícita, que o mesmo deveria ser preenchido apenas por doentes celíacos, com idade igual

ou superior a 16 anos, a residir em Portugal (eram solicitados dados relativos ao local de residência). Além disso, os participantes eram solicitados a responder apenas uma vez ao questionário, apesar de poderem tomar conhecimento do mesmo várias vezes. O questionário esteve online entre 18 de abril e 15 de julho de 2011. Neste período obtiveram-se 201 questionários válidos. Para este estudo em particular

foram apenas consideradas as respostas de 195 indivíduos, uma vez que os restantes 6 eram menores de 18 anos e o instrumento SF-36 encontra-se validado somente para adultos. Após a conclusão do período de recolha de dados as respostas foram extraídas da base de dados MySQL e convertidas para o software de análise estatística IBM SPSS Statistics®, versão 19.0. As variáveis categóricas foram descritas através de Venetoclax cost proporções, enquanto as variáveis contínuas foram descritas através de média e respetivo desvio-padrão (variáveis com distribuição normal) ou através de mediana e respetivo intervalo interquartil (variáveis que não apresentavam uma distribuição normal). A normalidade da distribuição das variáveis foi verificada através do teste de Kolmogorov-Smirnov. Foi considerado um nível de significância de 5%. Como apresentado na tabela 1, verifica-se que a maior parte dos participantes eram associados da APC (66,2%). A participação dos indivíduos do sexo feminino foi 8 vezes superior à dos indivíduos do sexo masculino (88,7 vs. 11,3%).

All other chemicals (e.g., acetic acid, sodium sulfate anhydrous, tetracycline,

cycloheximide, glucose and xylose) were of analytical grade and purchased from Sigma–Aldrich (USA). The Cellic CTec 2 cellulose enzyme was obtained from Novozyme (Canada). Experiments were conducted with a Leistritz co-rotating twin screw extruder (American Leistritz Extruder Corp, USA). The extruder was composed of twelve modular barrels that were each 200 mm long. The barrels were electrically heated using thermal induction and cooled by water circulation. Barrel temperature, water flow rate, feed flow rate and pressure were monitored from a control panel. The material was fed into the extruder inlet port (Barrel 0, Fig.

1) at 4 kg/h by a gravimetric feeder (Brabender Obeticholic Acid chemical structure Technology, Canada). Water was injected into Barrel 8 by a positive displacement pump (Milton Roy USA). A solid/liquid separator was positioned in Barrel 9 to collect the filtrate mainly containing dissolved xylose. Two pressure sensors were positioned in Barrels 8 and 10, respectively, to detect the pressure on both sides of the filter. Two screw configuration profiles (Fig. 1A and B) were used to produce the extruded corncobs with 7% and 80% xylose removals, respectively. These two screw configuration profiles were built by placing conveying, kneading and reverse screw elements at different positions and intervals. The conveying screw elements were used for material I-BET-762 in vivo transportation and their smaller pitch could Amobarbital compress the products and achieve a high degree of filling within each barrel. Kneading screw elements oriented at different angles were used to break down large solids and to mix biomass and water to achieve a homogeneous distribution. In addition, reverse screw

elements carrying the materials in the opposite direction were placed immediately before and after the filter to increase forward and backward pressure. The only differences between these two screw configuration profiles concerned their backward pressure development zones, situated in zone 11. The backward pressure development zone was composed of two reverse screw elements for Profile A, but only one for Profile B, which caused lower backward pressure, resulting in less xylose removal. All experiments were conducted at a barrel temperature of 100 °C, screw speed of 100 rpm, and a L/S ratio of 1.2. The concentration of glucose was quantified by an Agilent 1260 Infinity high-performance liquid chromatography (HPLC) using a MetaCarb H Plus Column 300 × 7.8 mm (Agilent Technologies, USA), equipped with a refractive index detector. Before analysis, hydrolyzed liquid samples were subjected to 50× dilutions and filtered through a 0.2 μm cellulose acetate membrane (VWR International, USA). The column temperature was maintained at 60 °C and the flow rate was 0.7 ml/min (5 mM H2SO4).

91 m ha (Central Water Commission, 2010). These reservoirs also support a wide variety of wildlife. Many of the reservoirs such as Govind Sagar Lake formed by diverting river Satluj (Bhakra Dam, Punjab) and Hirakud reservoir (Sambalpur, Orissa) are a major tourist attraction. As per official estimates, tourism contribution to India’s GDP and employment in 2007–2008 was 5.92% and 9.24% respectively (Government of India, 2012). These are very important numbers as wetlands (such as coral reefs, beaches, reservoirs, lakes and rivers) are considered

to be a significant part of the tourism experience and are likely to be a key part of the expansion in demand for Veliparib tourism locations (MEA, 2005 and Ramsar Convention on Wetlands and WTO, 2012). Every year, on an average nearly seven million tourist visit Kerala’s backwaters, beaches and wildlife sanctuaries; three million visit Uttarakhand’s lakes and other natural wetlands; one million visit Dal lake; and 20,000 visit lake Tsomoriri. In terms of growth in fish production in India, wetlands play a significant role. At the moment,

majority of fish production in the country is from inland water bodies (61% of total production), i.e. rivers; canals; reservoirs; tanks; ponds; and lakes (Table 2). It increased from 0.2 million tonne in 1950–1951 to about 5.1 million tonne in 2010–2011. Carp constitute about 80% of the total inland aquaculture production. Presently, the State of West Bengal occupies the topmost position (30% of total inland fish production) followed by Andhra Pradesh, Uttar Pradesh, Capmatinib research buy Bihar and Orissa (Ministry of Agriculture, 2012). Overall, fisheries accounts for 1.2% of India’s total Gross Domestic Product (GDP) and 5.4% of total agricultural GDP. Swamps, mangroves, peat lands, mires and marshes ADP ribosylation factor play an important role in carbon cycle. While wetland sediments are the long-term stores of carbon, short-term stores are in wetland existing biomass (plants, animals, bacteria and fungi) and dissolved components in the surface and groundwater (Wylynko, 1999). Though wetlands contribute about 40% of the global methane (CH4) emissions, they have the highest

carbon (C) density among terrestrial ecosystems and relatively greater capacities to sequester additional carbon dioxide (CO2) (Pant et al., 2003). Wetlands sequester C through high rates of organic matter inputs and reduced rates of decompositions (Pant et al., 2003). Wetland soils may contain as much as 200 times more C than its vegetation. However, drainage of large areas of wetlands and their subsequent cultivation at many places had made them a net source of CO2. Restoration of wetlands can reverse them to a sink of atmospheric CO2 (Lal, 2008). As per the estimations, carbon sequestration potential of restored wetlands (over 50 year period) comes out to be about 0.4 tonnes C/ha/year (IPCC, 2000). In India, coastal wetlands are playing a major role in carbon sequestration.

E., 1993. Water-balance of over-wintering beetles in relation to strategies for cold tolerance. Journal of Comparative Physiology B 163, 1–4. Olsvik, P.A., Gundersen, P., Andersen, R.A., Zachariassen, K.E., 2000. Metal accumulation and metallothionein in two populations of brown trout, Salmo trutta, exposed

to different natural water environments during a run-off episode. Aquatic Toxicology 50, 301–316. Pedersen, S.A., Kristiansen, E., Andersen, R.A., Zachariassen, K.E., 2007. Isolation and preliminary characterization of a Cd-binding protein from Tenebrio molitor (Coleoptera). Comparative Biochemistry and Physiology C 145, 457–463. Pedersen, S.A., Kristiansen, E., Andersen, R.A., Zachariassen, K.E., 2008. Cadmium is deposited in the gut content PD-L1 inhibitor of larvae of the beetle Tenebrio molitor and involves a Cd-binding protein of the low cysteine type. Comparative Biochemistry and Physiology C 148, 217–222. Pedersen, S.A., Zachariassen,

K.E., 2002. Sodium regulation during dehydration of a herbivorous and a carnivorous beetle from African ABT-199 molecular weight dry savannah. Journal of Insect Physiology 48, 925–932. Somme, L., Zachariassen, K.E., 1981. Adaptations to low-temperature in high-altitude insects from Mount Kenya. Ecological Entomology 6, 199–204. Zachariassen, K.E., 1979. Mechanism of the cryoprotective effect of glycerol in beetles tolerant to freezing. Journal of Insect Physiology 25, 29–32. Zachariassen, K.E., 1980. The role of polyols and nucleating-agents in cold-hardy beetles. Journal of Comparative Physiology 140, 227–234. Zachariassen, K.E., 1982. Nucleating-agents in cold-hardy insects. Comparative Biochemistry and Physiology A 73, 557–562. Zachariassen, K.E., 1985. Physiology of cold tolerance in insects. Physiological Reviews 65, 799–832. Zachariassen, K.E., 1989. Thermal adaptations to polar environments. In: Mercer, J.M. (Ed.), Thermal Physiology, Miconazole pp. 23–34. Zachariassen, K.E., 1991. Routes of transpiratory water-loss in a dry-habitat tenebrionid beetle. Journal of Experimental Biology 157, 425–437. Zachariassen, K.E., 1991. The water relations of overwintering insects. In: Lee, R.E.,

Denlinger, D. (Eds.), Insects at Low Temperature. Chapman and Hall, New York, pp. 47–63. Zachariassen, K.E., 1996. The water conserving physiological compromise of desert insects. European Journal of Entomology 93, 359–367. Zachariassen, K.E., Andersen, J., Kamau, J.M.Z., Maloiy, G.M.O., 1988. Water-loss in insects from arid and humid habitats in East-Africa. Acta Entomologica Bohemoslovaca 85, 81–93. Zachariassen, K.E., Andersen, J., Maloiy, G.M.O., Kamau, J.M.Z., 1987. Transpiratory water-loss and metabolism of beetles from arid areas in East-Africa. Comparative Biochemistry and Physiology A 86, 403–408. Zachariassen, K.E., Baust, J.G., Lee, R.E., 1982. A method for quantitative determination of ice nucleating-agents in insect hemolymph. Cryobiology 19, 180–184. Zachariassen, K.E., DeVries, A.L., Hunt, B., Kristiansen, E., 2002.

16 and 17 In more than 3000 patients treated to date, sofosbuvir has been shown to be safe, viral breakthrough during treatment has been rare (and associated with nonadherence), and few drug interactions

have been observed.18 and 19 In addition, patients with hepatic impairment do not require modification of sofosbuvir dosage.20 We conducted this open-label study to determine if the administration of up to 48 weeks of sofosbuvir and ribavirin to HCV-infected patients with liver cancer before liver transplantation could prevent post-transplant recurrence of HCV infection. Patients were enrolled at 13 centers in the United States, 1 in New Zealand, and 1 in Spain. Eligible patients were at least 18 years old with a body mass index selleck products of ≥18 kg/m2 and documented HCV infection of any genotype with an HCV-RNA value greater than 104 IU/mL. Patients who had failed previous treatment for HCV were eligible. Patients were required to be on the waiting list for liver transplantation (with anticipated time until transplantation of <1 y) from a deceased donor. Patients had HCC meeting the Milan criteria,21 with a biological model for end-stage liver disease (MELD) score of less Epigenetics inhibitor than 22, and a Child–Turcotte–Pugh score of 7 or less, and had to

be eligible for a MELD exception score as per the policy of the United Network for Organ Sharing. We chose to study patients with HCC because they could be expected to undergo liver transplantation within 1 year. This phase 2, open-label, pilot study had 2 phases: a pretransplant treatment phase and a post-transplant follow-up phase. During the pretransplant treatment phase, patients received sofosbuvir (Gilead Sciences) administered orally

at a dose of 400 mg once daily, along with ribavirin (Ribasphere; Kadmon) administered orally as a divided dose according to body weight (1000 mg/day in patients with a body weight of <75 kg, and 1200 mg/day in patients with a body weight of ≥75 kg). According to the original study protocol, treatment mafosfamide lasted up to 24 weeks or until time of transplant, whichever occurred first. Patients who completed treatment before transplantation also were assessed for sustained virologic response. Patients who relapsed after stopping the study drug during the pretransplant treatment phase and who were not found to have the S282T NS5B mutation (which is associated with resistance to sofosbuvir) were allowed to restart treatment and continue for an additional 24 weeks or until transplant. In a subsequent amendment, the study design was changed to allow all patients who had not reached 24 weeks of treatment at the time of the amendment to continue treatment uninterrupted to 48 weeks or transplant. This change was made after observing virologic relapse in 3 patients before transplantation in patients who stopped treatment after completing 24 weeks.

1A). In contrast, 1 h after venom injection (100 μg/100 μl) the muscles showed intense interstitial hemorrhage, edema (fiber enlargement and an increase in the tissue interfascicular space), and emaciated myofibers; neutrophils were very few or absent. At 3 h, some of the swollen edematous cells showed “delta” lesions and, more rarely, dense clumps of hypercontracted myofibrils; an inflammatory neutrophilic infiltrate was observed

along with extravascular red blood cells (Fig. 1B,C). At 6 h, hemorrhage diminished and was replaced by foci of a dense polymorphonuclear-rich Tyrosine Kinase Inhibitor Library price inflammatory infiltrate (not shown), while at 18 h macrophages gradually appeared in the interstitial space and in necrotic fibers (Fig. 1D). At 3 days post-venom, clusters of proliferative myoblasts were observed in the damaged area (Fig. 1E) and at 7 days post-venom basophilic myoblasts and myotubes of different sizes were abundant (Fig. 1F).

Doxorubicin manufacturer By 21 days post-venom, the former damaged region was now occupied by discrete foci of small centrally nucleated myofibers (shown in Fig. 3B). Masson’s trichrome, which stains collagen tissue blue, showed that in control muscle the collagenous matrix was clearly visible only in the perimysium around venous and arterial branches; the endomysial matrix was barely seen (Fig. 2A). At 1 h post-venom, the collagen bundles appeared ragged (Fig. 2B), while at 3 and 7 days post-venom there was a noticeable deposition in some regions, with proliferative myoblasts and differentiating myotubes giving these regions a fibrotic aspect (Fig. 2C,D, respectively). One hour after venom injection, the mean small diameter of fibers in the affected muscle region was significantly greater (59.8 ± 8.2 μm; n = 1200 fibers) than in controls injected with PBS (39.9 ± 6.7 μm; n = 1200 fibers) ( Fig. 3A). At 21 days post-venom, only 205 regenerated cells (those with central nuclei) were detected in cross-sections of gastrocnemius (n = 6 rats). The mean diameter of these fibers was significantly smaller (21.42 ± 5.65 μm) than that of apparently normal fibers

with peripheral nuclei in venom-injected muscle (41.16 ± 7.96 μm; Fig. 3B). The diameter of the latter fibers was not significantly different from Ribonuclease T1 that of normal fibers in rats injected with PBS (42.0 ± 6.03 μm). The activation of resident macrophages (M1 phagocytic population) in the damaged region was evaluated by counting the number of cells expressing CD68 protein in envenomed and control muscles; the number of CD68-positive macrophages increased significantly up to 24 h (461.67 ± 144.67), but decreased gradually thereafter. From 7 days post-venom onwards the number of CD68-positive cells was lower than observed at 1 h (Fig. 4A,B). Osteopontin was expressed in the cytoplasm of intact and venom-damaged fibers (Fig. 5A,B), macrophages, myoblasts and myotubes and fibroblast (Fig. 6 and Fig. 7).