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Objective  Translated and validated outcome instruments are of great importance, since they can be used for researchers studying different populations with the same problem. The objective of the present study was to translate, culturally adapt and validate the Hip Disability and Osteoarthritis Outcome Score (HOOS) into Brazilian Portuguese. Methods  The HOOS was translated from English into Brazilian Portuguese, translated back into English, and submitted to an experts committee. It was administered to 100 patients with hip osteoarthritis. The psychometric evaluation included factor analysis; internal reliability measures, test-retest reliability at 7 days, and construct validity comparison with the Brazilian version of the Graded Chronic Pain Scale (GCPS). Results  Factor analyses demonstrated a five-factor solution. The test-retest reliability showed a high degree of internal consistency for the five subscales ( pain and physical difficulties , 0.97 at baseline and 0.93 at 7 days; pain and difficulty sitting, lying down and getting up , 0.93 at baseline and 0.89 at 7 days; difficulty flexing the knee , 0.92 at baseline and 0.83 at 7 days; difficulty walking , 0.88 at baseline and 0.87 at 7 days; quality of life , 0.80 at baseline and 0.35 at 7 days). The construct validity was established during the comparison of the Brazilian version of the GCPS. Conclusions  A Brazilian version of the HOOS was developed with adequate reliability and validity. It will facilitate evaluation of the hip within a large patient population and across cultures.

Donepezil is one of the main compounds used in the therapy of Alzheimer's disease. Oral administration of this drug presents many drawbacks, resulting in treatment non-adherence among patients. Thus, the development of transdermal formulations for donepezil delivery is important. The aim of this study was to prepare and to evaluate nanostructured lipid carrier-based gels (NLC gel) able to improve the skin delivery of donepezil free base (DPB). The components of nanostructured lipid carriers (NLCs) were selected after evaluating their enhancing effects using in vitro DPB skin delivery assays. DPB-loaded NLC were prepared by a microemulsion technique, by employing stearic acid as a solid lipid, oleic acid as a liquid lipid, lecithin as a surfactant, and sodium taurodeoxycholate as a co-surfactant. The DPB-NLC dispersions were characterized morphologically using atomic force microscopy and physicochemically using dynamic light scattering and surface charge measurements. These data along, with the encapsulation studies, indicated that uniformly nano-sized particles with high drug encapsulation were fabricated. In vitro skin permeation assays were performed, and the results indicated that drug skin permeation from DPB-NLC gel was increased, not only by the enhancing effect of their components, but the lipid nanocarriers also presented an additional enhancing effect to increase drug flux across the skin. Therefore, DPB-NLC gel is an interesting formulation for the enhanced treatment of Alzheimer's disease., (Copyright © 2019. Published by Elsevier B.V.)

The treatment against leishmaniasis presents problems, since the currently used drugs are toxic and/or have high costs. In addition, parasite resistance has increased. As a consequence, in this study, a chloroquinolin derivative, namely 7-chloro-N,N-dimethylquinolin-4-amine or GF1059, was in vitro and in vivo tested against Leishmania parasites. Experiments were performed to evaluate in vitro antileishmanial activity and cytotoxicity, as well as the treatment of infected macrophages and the inhibition of infection using pre-treated parasites. This study also investigated the GF1059 mechanism of action in L. amazonensis. Results showed that the compound was highly effective against L. infantum and L. amazonensis, presenting a selectivity index of 154.6 and 86.4, respectively, against promastigotes and of 137.6 and 74.3, respectively, against amastigotes. GF1059 was also effective in the treatment of infected macrophages and inhibited the infection of these cells when parasites were pre-incubated with it. The molecule also induced changes in the parasites' mitochondrial membrane potential and cell integrity, and caused an increase in the reactive oxygen species production in L. amazonensis. Experiments performed in BALB/c mice, which had been previously infected with L. amazonensis promastigotes, and thus treated with GF1059, showed that these animals presented significant reductions in the parasite load when the infected tissue, spleen, liver, and draining lymph node were evaluated. GF1059-treated mice presented both lower parasitism and low levels of enzymatic markers, as compared to those receiving amphotericin B, which was used as control. In conclusion, data suggested that GF1059 can be considered a possible therapeutic target to be tested against leishmaniasis., (Copyright © 2019 Elsevier Inc. All rights reserved.)

New therapeutics against leishmaniasis are desirable, since the current drugs applied against this disease complex presents problems, such as the toxicity, high cost and/or parasite resistance. In the present study, a new fluoroquinoline derivate, namely 7-chloro-N-(4-fluorophenethyl)quinolin-4-amine or GF1061, was evaluated regarding to its in vitro antileishmanial action against Leishmania infantum and L. amazonensis species, as well as by its toxicity in mammalian cells and efficacy in the treatment of infected macrophages. The mechanism of action of this molecule in L. amazonensis and the therapeutic efficacy in infected BALB/c mice were also evaluated. Results showed that GF1061 was effective against both parasite species, showing selectivity index (SI) of 38.7 and 42.7 against L. infantum and L. amazonensis promastigotes, respectively, and of 45.0 and 48.9 against the amastigotes, respectively. Amphotericin B (AmpB), used as control, showed SI values of 6.6 and 8.8 against L. infantum and L. amazonensis promastigotes, respectively, and of 2.2 and 2.7 against the amastigotes, respectively. The molecule was effective in treat infected macrophages, as well as it induced alterations in the mitochondrial membrane potential, increase in the reactive oxygen species production, and in the cell integrity of the parasites. Regarding to the in vivo experiments, BALB/c mice (n = 8 per group) were subcutaneously infected with 10 6 L. amazonensis stationary promastigotes and, 60 days post-infection, they received saline or were treated during 10 days, once a day, with AmpB (1 mg/kg body weight) or GF1061 (5 mg/kg body weight). One day after the treatment, the infected tissue, spleen, liver, and draining lymph node (dLN) of the animals were collected, and the parasite load was evaluated. GF1061-treated mice, as compared to the saline and AmpB groups, showed significant reductions in the parasitism in the infected tissue (66% and 62%, respectively), liver (69% and 44%, respectively), spleen (71% and 38%, respectively), and dLN (72% and 48%, respectively). In conclusion, results suggested that GF1061 may be considered as a possible therapeutic target to be evaluated against leishmaniasis in other mammalian hosts., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Purpose: Tuberculosis (TB) treatment is often carried out empirically, based on clinical and radiological findings. Chest X-ray (CXR) has good sensitivity but poor specificity in TB diagnosis. Xpert MTB/RIF (Mycobacterium tuberculosis/Rifampicin) is increasingly used in many countries as the initial diagnostic test for TB. The aim of the present study was to evaluate the association of radiological findings with the Xpert MTB/RIF test in patients with suspected pulmonary TB., Methods: Cross-sectional study in an outpatient TB clinic. Sputum AFB smear, culture, Xpert MTB/RIF, and CXR were collected in patients with suspected pulmonary TB., Results: During the study period, 312 patients met the inclusion criteria and were included in the analysis. Among Xpert MTB/RIF-positive cases, the radiographic patterns were classified as typical of TB, compatible of TB, and normal in 78 (70.3%), 31 (27.9%), and 2 (1.8%) patients, respectively. CXRs were classified as typical of TB, compatible of TB, and normal in 20 (10.0%), 25 (12.4%), and 152 (75.6%) patients, respectively, in Xpert MTB/RIF-negative cases., Conclusions: We found an association between radiographic patterns and Xpert MTB/RIF results in patients with suspected pulmonary TB. These results confirm the current recommended diagnosis algorithm.

The evaluation of testicular hemodynamics can contribute significantly to the understanding of the thermoregulatory mechanisms and oxygen supply of the testis in domestic animals. The present study aimed to characterize circulatory dynamics using the mean velocity (MV), pulsatility index (PI) and resistive index (RI) of the supratesticular artery in bulls. We evaluated 334 bulls of five different breeds (Nelore, Hereford, Aberdeen Angus, Braford and Brangus) by performing a velocimetry analysis using Doppler ultrasonography. Data were compared by Welch's ANOVA, Games-Howell (post-hoc test) and Spearman correlation with a significance level of 5%. The overall MV of 12.14 ± 0.30 cm/s differed among breeds. In addition, we observed that Brangus bulls showed higher (P < 0.05) MV (16.28 ± 1.02 cm/s) compared to Nelore bulls (8.76 ± 0.40 cm/s). The RI had an overall mean of 0.41 ± 0.01 and differed among breeds. We observed higher (P < 0.05) RI values in Hereford (0.44 ± 0.01) compared to Brangus (0.36 ± 0.02) animals. Overall, the PI values (0.33 ± 0.01) did not differ (P > 0.05) among breeds. The correlation between the PI and RI (0.936; P < 0.001) was high and positive; however, the correlations were low and negative between MV and the PI (-0.228; P < 0.001) and between MV and the RI (-0.270; P < 0.001). We concluded that there are differences in the MV and RI of the bulls' supratesticular arteries among the different evaluated breeds. Moreover, the presented values attributed to blood flow dynamics can serve as parameters in future studies and can be used to identify alternative diagnostic tools for infertility or to understand issues of adaptability in bulls., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The genus Anopheles encompasses several species considered as vectors of human infecting Plasmodium. Environmental changes are responsible for behavior changes in these vectors and therefore the pattern of malaria transmission. To better understand the dynamics of malaria transmission, this study aimed at identify the species of adult anophelines found in a malaria endemic urban area of the Amazon region, Mâncio Lima, located in the Acre State Brazil. Using Shannon-type light traps installed at 11 collection points near fish ponds, a total of 116 anophelines were collected belonging to nine species. Anopheles darlingi Root 1926 and An. albitarsis s.l. Lynch-Arribalzaga 1878 were the most abundant and predominant species. Despite the low number of captured adult anophelines, the occurrence of An. darlingi throughout all urban area and the presence of secondary vectors reinforce the need of a permanent and continuous entomological surveillance.

This study evaluated the thermoregulation and spermatogenic changes by scrotal temperature gradient using infrared thermography in testicular compromised bulls. Bulls were insulated (n = 6) for 72 hr and control animals (n = 3) remained without insulation during all the experimental period. Seminal evaluation was performed prior, at insult removal and once per week for 13 consecutive weeks. Mean temperature gradient in insulated animals was lower at the time of insulation removal compared to the week prior and after the insult (p < .05). Two weeks after insult, sperm motility was lower in insulated compared to control animals (p < .01) and spermatozoa total defects were higher in insulated compared to control animals (p < .05). Two and seven weeks after insult, the major defects were higher in insulated compared to control animals (p < .05). Scrotal temperature gradient showed a positive correlation with sperm mass motion (p < .01) and a negative correlation with ocular globe temperature (p < .01) in insulated animals. The infrared thermography can be used to evaluate ocular globe temperature in bulls; however, it is only effective to detect changes in scrotal temperature gradient at the insult removal., (© 2017 Blackwell Verlag GmbH.)

Background: Xpert MTB/RIF is increasingly used in many countries as the initial diagnostic test for tuberculosis (TB). Few studies have evaluated the effect of Xpert on TB diagnosis under programmatic conditions in Brazil. The aim of the present study was to evaluate the impact of introduction of Xpert MTB/RIF on TB diagnosis in a city with high TB incidence in Brazil., Methods: We included patients evaluated with conventional diagnostic tests during one year before Xpert introduction (pre-Xpert group) and patients evaluated using Xpert during one year after the test introduction (post-Xpert group)., Results: 620 patients met the inclusion criteria (208 in the pre-Xpert group and 412 in the post-Xpert group) and were included in the analysis. The time until TB diagnosis was shorter in post-Xpert group (0.7 day, IQR: 0.5-1.0 day) than in pre-Xpert group (2.0 days, IQR: 2.0-2.0 days) (p<0.0001). Atypical disease characteristics, such as less weight loss, fever, dyspnea, night sweats, and hemoptysis; a negative sputum smear; a negative culture, and a chest X-ray atypical of TB were more common in post-Xpert group than in pre-Xpert group (p<0.0001 for all)., Conclusions: We found that the implementation of the Xpert MTB/RIF assay, under programmatic conditions, improve and facilitate TB diagnosis, especially in cases with atypical disease manifestations. These results are likely to be generalizable to settings with a similar high TB incidence.

Background: Lactose intolerance is characterized by the absence of the enzyme lactase (beta-galactosidase) and affects two thirds of the world adult population. Our aim was to evaluate a lactase gastro-resistant formulation to ensure increased activity in the action site of the enzyme (lumen of the small intestine). Simultaneously, we also evaluated the commercial product stability and enzyme activity, because the product containing beta-galactosidase is classified as food supplement according to the Food and Drug Administration (FDA), so it is free to pass quality testing, efficacy and stability. So, it is possible that contain some irregularities as to the content and enzymatic activity., Methods: The dissolution assay was performed using a dissolution test system and commercial product and the gastro-resistant formulation were evaluated according to a method adapted to the conditions recommended by United States Pharmacopeia (US Pharmacopeia) for gastro-resistant formulations. For the assessment of enzymatic activity throughout the dissolution test was employed the official method of lactase assay described in US Pharmacopoeia. This method is based on a colorimetric reaction which the substrate reacts with the enzyme generate a colored product further analyzed by UVVisible spectrophotometry., Results: When carrying out dissolution test in commercial product it is noted that the existing formulation is not able to protect the enzyme from degrading action of gastric environment (a loss of 86.0 ± 0.8% of lactase activity was observed). Our proposed gastro-resistant pharmaceutical form there was no loss of activity during the acid step and the end of the dissolution test the found activity was 95 ± 1.3%., Conclusion: The formulations proposed in this work using hypromellose capsules ensure the enzymatic activity of lactase, preventing its contact with the acid medium. For the other side, the results of commercial tablets for lactase release indicate a significant loss of enzyme activity due to the immediate release of the enzyme in the simulated gastric fluids., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.)

Lapachol is an abundant prenyl naphthoquinone occurring in Brazilian Bignoniaceae that was clinically used, in former times, as an antimalarial drug, despite its moderate effect. Aiming to search for potentially better antimalarials, a series of 1,2,3-triazole derivatives was synthesized by chemical modification of lapachol. Alkylation of the hydroxyl group gave its propargyl ether which, via copper-catalyzed cycloaddition (CuAAC) click chemistry with different organic azides, afforded 17 naphthoquinonolyl triazole derivatives. All the synthetic compounds were evaluated for their in vitro activity against chloroquine resistant Plasmodium falciparum (W2) and for cytotoxicity to HepG2 cells. Compounds containing the naphthoquinolyl triazole moieties showed higher antimalarial activity than lapachol (IC 50 123.5 μM) and selectivity index (SI) values in the range of 4.5-197.7. Molecular docking simulations of lapachol, atovaquone and all the newly synthesized compounds were carried out for interactions with PfDHODH, a mitochondrial enzyme of the parasite respiratory chain that is essential for de novo pyrimidine biosynthesis. Docking of the naphthoquinonolyl triazole derivatives to PfDHODH yielded scores between -9.375 and -14.55 units, compared to -9.137 for lapachol and -12.95 for atovaquone and disclosed the derivative 17 as a lead compound. Therefore, the study results show the enhancement of DHODH binding affinity correlated with improvement of SI values and in vitro activities of the lapachol derivatives., (Copyright © 2017 Elsevier Masson SAS. All rights reserved.)

Although boar semen productivity is affected by seasonality, its effects are not equal among different regions which raise concerns regarding the profitability of boar stud farms. Therefore, the goals of this study were (i) to evaluate the seasonal effect on semen production in a commercial boar stud farm located in a subtropical climate region and (ii) to verify whether the activities of superoxide dismutase and glutathione peroxidase in spermatozoa and seminal plasma were associated with seminal traits of fresh and cooled semen. Nine boars were collected twice per season, and routine seminal parameter analyses were performed together with superoxide dismutase and glutathione peroxidase activities in seminal plasma and spermatozoa. Despite a reduction in sperm concentration in spring and summer, most seminal parameters were constant year-round. Temperature-humidity index was higher in the summer compared to spring, autumn and winter (p < .05). Superoxide dismutase activity in spermatozoa was increased in summer compared to autumn and winter (p < .05). The activities of both enzymes in seminal plasma and spermatozoa glutathione peroxidase remained unaltered throughout the seasons. In conclusion, seasonality showed little influence in overall boar seminal parameters despite microclimatic differences among seasons, and spermatozoa collected during summer increased superoxide dismutase activity., (© 2018 Blackwell Verlag GmbH.)

Background and Objectives: As for all tests, the diagnostic performances of Xpert MTB/RIF might be different in settings with different tuberculosis prevalence. Aim of the study is to evaluate the performances of Xpert MTB/RIF to diagnose tuberculosis in Brazil, where 407 culture-confirmed tuberculosis patients were retrospectively enrolled in Rio Grande do Sul, between 2015 and 2016., Methods: Sensitivity, specificity, positive and negative predictive values of the test were calculated and a logistic regression analysis was performed to assess the role played by explanatory variables in the occurrence of true positive and negative diagnostic results., Results: Sensitivity of Xpert MTB/RIF was 100.0%, specificity 92.8%; positive and negative predictive values were 71.4% and 100.0%, respectively. In the HIV- infected sub-group specificity was 59.3%. In the multivariate logistic regression analysis, true positivity was associated with increasing age (1.0; p-value: 0.02) while true positivity and negativity were negatively associated with alcohol abuse., Conclusions: Xpert is sensitive and specific in the Brasilian settings., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The objective of this study was to compare the presence of ovarian activity and pregnancy rates to temperature variation at the vulvar skin measured by infrared thermography (IRT). In addition, we also aimed to evaluate the IRT as a non-invasive method to evaluate animal breeding from fixed timed artificial insemination (FTAI). The study comprises 150 non-lactating beef Braford cows (5/8 Hereford × 3/8 Nellore) aged between 3 and 10 years. Data were collected along the FTAI protocol period during animal management. Animals were subjected to reproductive ultrasound evaluation and thermal images were performed by an infrared camera. Mean skin vulvar temperature (°C) and ovarian structures data were compared using Tukey's t test used as follow-up test to ANOVA. We observed a statistical difference in the mean vulvar skin temperature between animals that had the presence of ovarian follicles (34.2 ± 1.8) compared to no activity (35.4 ± 1.0; P < 0.05). However, vulvar skin temperature were similar between pregnant (34.5 ± 1.5) compared to non-pregnant (34.3 ± 1.9) animals (P > 0.05). In conclusion, the IRT technique was efficient to detect changes on vulvar skin temperature observed during FTAI protocol in Braford cows. Therefore, the use of IRT as an indirectly diagnostic tool to detect ovarian activity seems promising and further studies are required to validate their potential in beef cattle production.

This study was conducted on 78 13-month-old crossbred beef heifers that weighed 215 kg in Southern Rio Grande do Sul (RS) State, Brazil. We evaluated the performance of beef heifers that were reared in a pasture system that received a mineral supplement energy-type protein with added sodium lasalocid (LAS). The heifers were randomly and uniformly divided into 2 groups, with 39 animals in each group. One group of animals received a mineral supplement energy-type protein without sodium lasalocid (CON), and the other group received a mineral supplement energy-type protein with added LAS. The mean feed intake, the body weight (BW), the average daily gain (ADG), the body condition score (BCS), and ovarian cyclicity were recorded, and economic parameters were calculated. No differences in supplement intake were observed between the groups, which ensures adequate intake of the other components of the mineral mixture, which are part of the nutritional requirements for the production process. Similarly, no difference in the ADG was observed between treatments. We observed that the heifers in the LAS group had a higher BW gain (51 kg) that the CON heifers (40 kg; P < 0.05). In addition, LAS-supplemented heifers had a higher BCS (3.53) than CON heifers (3.38) at the end of the experiment (P < 0.05). The heifers supplemented with LAS had a higher profitability than the CON heifers, even with the higher cost of the supplement containing LAS; this effect was due to the higher live BW at the end of the study. We concluded that the administration of a mineral supplement energy-type protein with added LAS has beneficial effects on beef heifers in terms of production and economic feasibility.

The aim of this study was to evaluate the bioclimatic thermal stress assessed by Equivalent Temperature Index (ETI) and Temperature Humidity Index (THI) on Braford and Nellore bulls sperm quality during the reproductive seasons at the tropical region in the Brazilian Pantanal. We used 20 bulls aged approximately 24 months at the beginning of the study. Five ejaculates per animal were collected using an electroejaculator. Temperature, air humidity, and wind speed data were collected every hour from the automatic weather station at the National Institute of Meteorology. Infrared thermography images data were collected to assess the testicular temperature gradient in each animal. Data were analyzed with ANOVA using MIXED procedure of SAS and means were compared using Tukey's HSD test. The THI and ETI at 12 days (epididymal transit) were higher in January (89.7 and 28.5, respectively) and February (90.0 and 29.0, respectively) compared to other months (P < 0.01). Total seminal defects differ only in Bradford bulls between the months of November and February. Nellore bulls had lower major defects (MaD) and total defects (TD) compared to Braford. Nellore bulls showed correlation between minor defects (MiD) and THI for 30 days (0.90) and 18 days (0.88; P < 0.05). Braford bulls showed correlation for MaD (0.89) in ETI for 12 days (P < 0.05). Infrared thermography showed no difference between animals. Reproductive response to environmental changes is a consequence of Nellore and Braford adaptation to climate stress conditions. Both THI and ETI environmental indexes can be used to evaluate the morphological changes in the seminal parameters in Nellore or Braford bulls; however, more experiments should be performed focusing on larger sample numbers and also in reproductive assessment during the consecutive years to assess fertility potential.

Donepezil is a drug administered for Alzheimer's disease treatment, and it is a potential template molecule for imprinted microparticles. The precipitation polymerization technique allows the synthesis of spherical imprinted microparticles, and the intermolecular interactions among drug and molecularly imprinted polymers (MIPs) play a promising role for delineating drug delivery systems. Once that donepezil is a poorly-water soluble compound, lipid based-formulations (LBFs) may enhance its oral administration. Based on this, LBFs are useful vehicles to incorporate imprinted microparticles synthesized by precipitation polymerization. In these formulations, the drug dissolved in lipids is accessible to adsorbate in the polymers, and the hydrophobic environment of lipids increases the molecular recognition of MIPs. The formulations based on MIPs using pure oleic acid as vehicle prolong the in vitro release of donepezil up to several hours by a Fickian diffusion mechanism, and it provides a multiphasic release pattern related to the heterogeneity of the binding sites. The modulation of donepezil release from MIPs-based formulations using oil vehicles may contribute to decrease its side effects, possibly regulating its absorption rate in the gastrointestinal tract. These systems represent a novel technological platform to prolong the delivery not only for donepezil, but also for a variety of therapeutics., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Data structures have been explored for several domains of computer applications in order to ensure efficiency in the data store and retrieval. However, data structures can present different behavior depending on applications that they are being used. Three-dimensional interactive environments offered by techniques of Virtual Reality require operations of loading and manipulating objects in real time, where realism and response time are two important requirements. Efficient representation of geometrical models plays an important part so that the simulation may become real. In this paper, we present the implementation and the comparison of two topologically efficient data structures - Compact Half-Edge and Mate-Face - for the representation of objects for three-dimensional interactive environments. The structures have been tested at different conditions of processors and RAM memories. The results show that both these structures can be used in an efficient manner. Mate-Face structure has shown itself to be more efficient for the manipulation of neighborhood relationships and the Compact Half-Edge was more efficient for loading of the geometric models. We also evaluated the data structures embedded in applications of biopsy simulation using virtual reality, considering a deformation simulation method applied in virtual human organs. The results showed that their use allows the building of applications considering objects with high resolutions (number of vertices), without significant impact in the time spent in the simulation. Therefore, their use contributes for the construction of more realistic simulators., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Previous studies have shown that the negative effects of density of Ae. albopictus on Ae. aegypti exceed those of Ae. aegypti on Ae. albopictus for population growth, adult size, survivorship, and developmental rate. This competitive superiority has been invoked to explain the displacement of Ae. aegypti by Ae. albopictus in the southeastern USA. In Brazil, these species coexist in many vegetated suburban and rural areas. We investigated a related, but less-well-studied question: do effects of Ae. albopictus on Ae. aegypti larval development and survival occur under field conditions at realistic densities across multiple seasons in Brazil? We conducted additive competition experiments in a vegetated area of Rio de Janeiro where these species coexist. We tested the hypothesis that Ae. aegypti (the focal species, at a fixed density) suffers negative effects on development and survivorship across a gradient of increasing densities of Ae. albopictus (the associate species) in three seasons. The results showed statistically significant effects of both season and larval density on Ae. aegypti survivorship, and significant effects of season on development rate, with no significant season-density interactions. Densities of Aedes larvae in these habitats differed among seasons by a factor of up to 7x. Overall, Spring was the most favorable season for Ae. aegypti survivorship and development. Results showed that under natural conditions the negative competitive effects of Ae. albopictus on Ae. aegypti were expressed primarily as lower survivorship. Coexistence between Ae. aegypti and Ae. albopictus in vegetated areas is likely affected by seasonal environmental differences, such as detrital resource levels or egg desiccation, which can influence competition between these species. Interactions between these Aedes are important in Brazil, where both species are well established and widely distributed and vector dengue, Zika and chikungunya viruses.

Objective: To compare the activation of the vastus lateralis (VL) and biceps femoris (BF) muscles during gait, as well VL/BF muscular co-contraction (MCC) between healthy (CG) and anterior cruciate ligament reconstructed (ACL-R) subjects., Methods: Nineteen subjects, ten controls and nine ACL-R patients had a VL and BF electromyogram (EMG) captured to calculate the MCC ratio. A Principal Component (PC) Analysis was applied to reduce the dimensionality effect of each of the MCC, VL and BF curves for both healthy and ACL reconstructed groups. The PC scores were used to calculate the standard distance (SD). SD values were employed in order to compare each dependent variable (MCC, VL and BF) between the two groups using unpaired t-test., Results: ACL-R group presented a lower VL activation at the beginning and at the end of the gait cycle, as compared to the control group. However, no difference was found for BF or VL/BF MCC., Conclusion: The gait analysis of ACL reconstructed patients demonstrated a persistent deficit in VL activation when compared to the control group, even one year after surgery. Level of Evidence III. Case Control Study.

Objective: : To compare gait spatiotemporal parameters of healthy and ACL reconstructed subjects in order to classify the status of gait normality., Methods: : Fourteen healthy subjects and eight patients submitted to ACL reconstruction walked along a walkway while the lower limbs movement was captured by an infrared camera system. The frames where the initial contact and toe-off took place were determined and the following dependent variables, which were compared between groups through the Mann-Whitney test (a=0.05) were calculated: percentage of time in initial double stance, percentage of time in single stance, percentage of time in terminal double stance, stride length and gait velocity. Initially, all variables were compared between groups using a Mann-Whitney test. A logistic regression was applied, including all dependent variables, to create a model that could differentiate healthy and ACL reconstructed subjects., Results: : ACL reconstructed group showed no differences in any spatiotemporal parameter of gait (p > 0.05) in relation to the control group, although the angular kinematic differences of the knee remained altered, as evidenced in a study with a similar sample., Conclusion: : The regression classified all subjects as healthy, including the ACL reconstructed group, suggesting the spatiotemporal variables should not be used as the sole criterion of return to sports activities at the same level as prior to injury. Level of Evidence III, Case Control Study.

The objective of this study was to evaluate the seasonal effects of the environment on sperm quality in subtropical region determined by temperature and humidity index (THI). We used 20 Brangus bulls (5/8 Angus × 3/8 Nellore) aged approximately 24 months at the beginning of the study. Semen evaluations were performed twice per season during 1 year. Climate THI data were collected from an automatic weather station from the National Institute of Meteorology. Infrared thermography images were used to determine the temperature of the proximal and distal poles of the testis to assess the testicular temperature gradient (TG). The seasonal effects on seminal and climatic variables were analyzed with ANOVA using MIXED procedure of SAS. Sperm motility in spring (60.1%), summer (57.6%), and autumn (64.5%) showed difference compared to winter (73.0%; P < 0.01). TG was negatively correlated with THI at 18 days (spermiogenesis) (-0.76; P < 0.05) and at 12 days (epididymal transit) (-0.85; P < 0.01). Ocular temperature (OcT) had a positive correlation with THI at 18 days (0.78; P < 0.05) and at 12 days (0.84; P < 0.01). Motility showed a negative correlation with THI only at 18 days (-0.79; P < 0.05). During spermiogenesis, the TG had higher negative correlation compared to OcT (-0.97; P < 0.01) and rectal temperature (-0.72; P < 0.05). Spermatozoa with distal midpiece reflex were correlated with THI during transit epididymis (0.72; P < 0.05). Seminal parameters are not affected when THI reaches 93.0 (spermiogenesis) and 88.0 (epididymal transit). We concluded that infrared thermography can be adopted as an indirect method in order to assess the effect of environmental changes in TG and OcT of Brangus bulls.

Donepezil is a drug usually administered by oral route for Alzheimer disease treatment, but several gastric side effects have been reported as diarrhea, nausea, and anorexia. We explored the phase behavior of lyotropic liquid crystalline (LLC) mesophases composed by monoolein/oleic acid/water for enhanced administration of donepezil. Polarized light microscopy suggested that these systems ranged from isotropic inverse micellar solutions (L2) to viscous and birefringent reverse hexagonal (HII) mesophases according to the amount of water in the ternary systems. Phase transition was observed from a L2 phase to HII mesophase after swelling studies, an interesting property to be explored as a precursor of LLC mesophases for mucosal administration that increases its viscosity in situ. Mucoadhesive properties of LLC mesophases were characterized using a texture analyzer indicating that these systems can have an increased residence time in the site of absorption. Donepezil-free base was incorporated in the evaluated formulations, and their in vitro release was controlled up to 24 h. The phase behavior of the systems demonstrated a great potential for enhanced donepezil administration once these mucoadhesive-controlled release formulations can incorporate the drug and prolong its release, possibly reducing its side effects., (Copyright © 2016 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.)

The objectives of this study were firstly to determine whether the stimulatory function of equine growth hormone (eGH) on equine oocyte maturation in vitro is mediated via cyclic adenosine monophosphate (cAMP); and secondly if the addition of eGH in vitro influences oocyte nuclear maturation and if this effect is removed when GH inhibitors are added to the culture. Cumulus-oocyte complexes (COCs) were recovered from follicles <25 mm in diameter and randomly allocated as follows: (i) control (no additives); and (ii) 400 ng/ml of eGH. A specific inhibitor against cyclic AMP-dependent protein kinase (H-89; 10-9, 10-11 or 10-15 M concentration) and a specific adenylate cyclase inhibitor, 2',3'-dideoxyadenosine (DDA; 10-8, 10-10 or 10-14 M concentration) were used to observe whether they could block the eGH effect. After 30 h of in vitro maturation at 38.5°C with 5% CO2 in air, oocytes were stained with 10 μg/ml of Hoechst to evaluate nuclear status. More mature oocytes (P < 0.05) were detected when COCs were incubated with eGH (29 of 84; 34.5%) than in the control group (18 of 82; 21.9%). The H-89 inhibitor used at a concentration of 10-9 M (4 of 29; 13.8%) decreased (P < 0.05) the number of oocytes reaching nuclear maturation when compared with eGH (11 of 29; 38%). The DDA inhibitor at a concentration of 10-8 M (2 of 27; 7.4%) also reduced (P < 0.05) the number of oocytes reaching maturity when compared with the eGH group (9 of 30; 30%). Results from the present study show that H-89 and DDA can be used in vitro to block the eGH effect on equine oocyte maturation.

Background: At present, dengue control focuses on reducing the density of the primary vector for the disease, Aedes aegypti, which is the only vulnerable link in the chain of transmission. The use of new approaches for dengue entomological surveillance is extremely important, since present methods are inefficient. With this in mind, the present study seeks to analyze the spatio-temporal dynamics of A. aegypti infestation with oviposition traps, using efficient computational methods. These methods will allow for the implementation of the proposed model and methodology into surveillance and monitoring systems., Methods: The study area includes a region in the municipality of Rio de Janeiro, characterized by high population density, precarious domicile construction, and a general lack of infrastructure around it. Two hundred and forty traps were distributed in eight different sentinel areas, in order to continually monitor immature Aedes aegypti and Aedes albopictus mosquitoes. Collections were done weekly between November 2010 and August 2012. The relationship between egg number and climate and environmental variables was considered and evaluated through Bayesian zero-inflated spatio-temporal models. Parametric inference was performed using the Integrated Nested Laplace Approximation (INLA) method., Results: Infestation indexes indicated that ovipositing occurred during the entirety of the study period. The distance between each trap and the nearest boundary of the study area, minimum temperature and accumulated rainfall were all significantly related to the number of eggs present in the traps. Adjusting for the interaction between temperature and rainfall led to a more informative surveillance model, as such thresholds offer empirical information about the favorable climatic conditions for vector reproduction. Data were characterized by moderate time (0.29 - 0.43) and spatial (21.23 - 34.19 m) dependencies. The models also identified spatial patterns consistent with human population density in all sentinel areas. The results suggest the need for weekly surveillance in the study area, using traps allocated between 18 and 24 m, in order to understand the dengue vector dynamics., Conclusions: Aedes aegypti, due to it short generation time and strong response to climate triggers, tend to show an eruptive dynamics that is difficult to predict and understand through just temporal or spatial models. The proposed methodology allowed for the rapid and efficient implementation of spatio-temporal models that considered zero-inflation and the interaction between climate variables and patterns in oviposition, in such a way that the final model parameters contribute to the identification of priority areas for entomological surveillance.

We investigated the differences between weaning rates and technologies adopted by farmers in cow-calf production systems in Rio Grande do Sul State, Brazil. Interviews were carried out with 73 farmers about 48 technologies that could affect reproductive performance. Data were analyzed by multivariate analysis using a non-hierarchical cluster method. The level of significance was set at P < 0.05. Three distinct clusters of farmers were created (R (2) = 0.90), named as low (LWR), intermediate (IWR), and high (HWR) weaning rate, with 100, 91, and 96 % of the farmers identified within their respective groups and average weaning rates of 59, 72, and 83 %, respectively. IWR and HWR farmers used more improved natural pasture, fixed-time artificial insemination, selection for birth weight, and proteinated salt compared to LWR. HWR farmers used more stocking rate control, and IWR farmers used more ultrasound to evaluate reproductive performance compared to the LWR group. IWR and HWR adopted more technologies related to nutrition and reproductive aspects of the herd in comparison to LWR. We concluded that farmers with higher technology use on farm had higher weaning rates which could be used to benefit less efficient farmers.

Rheum emodi is principally known to consist 1,8-dihydroxyanthraquinones (DHAQs) that find immense use in the chemical, pharmaceutical, cosmetic industries and in herbal medication and food sector. The aim of this study was to compare non-conventional and classical methods for extraction of anthraquinones from R. emodi. Optimisation of the extraction parameters for various methods was done and their extraction efficiency was evaluated. In preliminary screening experiments, choice of solvent and solid : solvent ratio was optimised. Comparison of extraction efficiency for classical methods like maceration, heat-reflux, soxhletion and non-conventional methods like ultra-sonication and sublimation was done for five DHAQs - aloe emodin, rhein, emodin, chrysophanol and physcion using HPLC-UV and fluorescence detection in native and acid hydrolysed samples. It was observed that ethanol was the best solvent for extraction of anthraquinones with a solid : solvent ratio of 1:20. A prior acid hydrolysis led to significant increase in anthraquinone extraction. Among the extraction methods heat reflux for 45 min was the most prominent extraction method with highest recovery of the DHAQs. In ultrasonic assisted extraction, an increase in the anthraquinone extraction was seen till 45 min after which the concentration declined. A novel, solvent-free, green and selective method of extraction by sublimation was found to be effective for extraction of anthraquinones.

Fish farming in the Amazon has been stimulated as a solution to increase economic development. However, poorly managed fish ponds have been sometimes associated with the presence of Anopheles spp. and consequently, with malaria transmission. In this study, we analyzed the spatial and temporal dynamics of malaria in the state of Acre (and more closely within a single county) to investigate the potential links between aquaculture and malaria transmission in this region. At the state level, we classified the 22 counties into three malaria endemicity patterns, based on the correlation between notification time series. Furthermore, the study period (2003-2013) was divided into two phases (epidemic and post-epidemic). Higher fish pond construction coincided both spatially and temporally with increased rate of malaria notification. Within one malaria endemic county, we investigated the relationship between the geolocation of malaria cases (2011-2012) and their distance to fish ponds. Entomological surveys carried out in these ponds provided measurements of anopheline abundance that were significantly associated with the abundance of malaria cases within 100 m of the ponds (P < 0.005; r = 0.39). These results taken together suggest that fish farming contributes to the maintenance of high transmission levels of malaria in this region.

Introduction: This study was based on the hypothesis that some components of the angiotensin-(1-7) (Ang-(1-7)) system are differentially expressed during follicular development and can be involved in the follicular health/atresia transition in bovine., Material and Methods: The largest (F1) and second largest follicles (F2) were collected from cows before (Day 2), during (Day 3), or after (Day 4) the expected moment of follicular deviation. In the second experiment, F1 was induced to atresia through intrafollicular injection of fulvestrant (estrogen receptor-antagonist) and, in both experiments, mRNA expression of the Mas receptor, ACE2, NEP, and PEP was evaluated in the granulosa and theca cells., Results: The mRNA expression of Mas receptor was upregulated in the granulosa cells of F2 after the establishment of follicular deviation, while PEP mRNA increased during and after the deviation process. The mRNA expression of ACE2 was upregulated in the granulosa cells of F1 during and after the follicular deviation. The mRNA expression of NEP was not regulated in F1 and F2. Mas receptor expression increased in the F1 induced to atresia., Conclusions: mRNA for Mas receptor, ACE2, and PEP are differentially expressed in granulosa cells throughout follicular development and the Mas receptor can be involved with the establishment of follicular dominance., (© The Author(s) 2013.)

The multidrug-resistant bacteria have become a serious problem to public health. In this scenery the antimicrobial peptides (AMPs) derived from animals and plants emerge as a novel therapeutic modality, substituting or in addition to the conventional antimicrobial. The anurans are one of the richest natural sources of AMPs. In this work several cycles of cDNA cloning of the skin of the Brazilian treefrog Hypsiboas semilineatus led to isolation of a precursor sequence that encodes a new AMP. The sequence comprises a 27 residue signal peptide, followed by an acidic intervening sequence that ends in the mature peptide at the carboxy terminal. The AMP, named Hs-1, has 20 amino acids residues, mostly arranged in an alpha helix and with a molecular weight of 2144.6 Da. The chemically synthesized Hs-1 showed an antimicrobial activity against all Gram-positive bacteria tested, with a range of 11-46 μM, but it did not show any effect against Gram-negative bacteria, which suggest that Hs-1 may have a selective action for Gram-positive bacteria. The effects of Hs-1 on bacterial cells were also demonstrated by transmission electron microscopy. Hs-1 is the first AMP to be described from H. semilineatus., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Introduction/objective: We evaluated the predictability of early changes in serum albumin (sAlb) on the two-year mortality of incident hemodialysis patients., Methods: Observational, longitudinal retrospective study using the database of Fresenius Medical Care of Latin America. Adult patients starting dialysis from January/2000 to June/2004, from 25 centers were included. Changes in sAlb during the first 3 months on hemodialysis were used as the main predictor. The outcome was death from any cause., Results: 1,679 incident patients were included. They were 52 ± 15 years old, 58.7% male and 21.5% diabetic, with a median sAlb of 38 g/L (bromocresol green). 923 patients had sAlb < 38 g/L (Low sAlb Group) and 756 ones had sAlb > 38.0 g/L (Adequate sAlb Group). The mortality was significantly higher in Low sAlb Group (17% vs. 11%, p < 0.001). Early changes in sAlb significantly affected two-year mortality. Factoring the Kaplan Meier curve of Low sAlb Group by the presence of an increase in sAlb uncovered of a statistically significant difference in mortality favoring the ones whose sAlb went up (19% vs. 15%, p = 0.043). Differently, patients from Adequate sAlb Group with a decrease in their sAlb had a statistically higher mortality rate (13% vs. 8%, p = 0.029)., Conclusions: Early sAlb changes showed a significant predictive power on mortality at 2 years in incident hemodialysis patients. Those with low initial sAlb may have a better prognosis if their sAlb rises. In contrast, patients with satisfactory initial levels can have a worsening of their prognosis in the case of an early reduction in sAlb.

The aim of this study was to assess the seasonal effects of the environment on semen quality in bulls, using infrared thermography. Sperm motility (M), mass motion (MM), and vigor (VIG) were evaluated in sperm samples from 17 Bradford bulls aged approximately 24 months at the beginning of the study. Infrared thermography images and data were collected using an infrared FLIR T 300 camera and Quick Report 1.2 SP2 software to determine the temperature of the proximal and distal poles of the testis and to assess the testicular temperature gradient. The seasonal effects on physiological, seminal, and climatic variables were analyzed by the GLM ANOVA and CORR procedures using SAS®. The microclimatic factors were recorded in hourly intervals, and the daily mean temperature and mean relative humidity were calculated to determine the daily temperature-humidity index (THI) every day for 1 year. The temperature gradient (TG) variations of the testes were significantly higher in the autumn (4.5 °C), winter (4.0 °C), and spring (2.9 °C) compared to summer (0.9 °C) (P < 0.05). Ocular globe temperatures were lower in the winter (27.6 °C) and autumn (26.8 °C) compared to summer (33.9 °C) and spring (31.1 °C) (P < 0.05). The average MM (2.58), M (52.64), and VIG (2.70) of the semen decreased in the summer compared to other seasons (P < 0.01). The TG was negatively correlated with THI (-0.44; P < 0.05). For the seminal variables, MaD (-0.45; P < 0.05) and TD (-0.50; P < 0.01) presented a negative correlation with TG. The TG had a positive correlation between M and VIG, which had values of 0.36 and 0.35, respectively (P < 0.05). We have concluded that infrared thermography can be used to assess the testicular temperature gradient and its consequences on physical and quantitative aspects of sperm.

Immature oocytes synthesize a variety of proteins that include the enzyme glucose-6-phosphate dehydrogenase (G6PDH). Brilliant cresyl blue (BCB) is a vital blue dye that assesses intracellular activity of G6PDH, an indirect measure of oocyte maturation. The objective was to evaluate the BCB test as a criterion to assess developmental competence of equine oocytes and to determine if equine growth hormone (eGH) enhanced in vitro maturation (IVM) of equine oocyte. Cumulus-oocytes complexes (COCs) were recovered by aspirating follicles <30 mm in diameter from abattoir-derived ovaries and were evaluated morphologically. Thereafter, COCs were exposed to BCB (26 μM) for 90 min at 39°C and selected based on the colour of their cytoplasm (BCB positive/BCB+ or BCB negative/BCB-). The COCs were allocated as follows: (a) IVM medium; (b) eGH group; (c) BCB-/IVM; (d) BCB+/IVM; (e) BCB-/eGH; and (f) BCB+/eGH. Then, COCs were cultured in vitro for 30 h, at 39°C in a 5%CO2 humidified air atmosphere. Cumulus-free oocytes were incubated in 10 μg/ml of bis-benzamide for 20 min at 39°C and nuclear maturation was evaluated with epifluorescence microscopy. Of the 39 COCs selected morphologically and subjected to BCB staining, 18/39 (46.2%) were classified as BCB+ and 21/39 (53.8%) as BCB- (P > 0.05). Maturation was not affected significantly by BCB classification, but the maturation rate was higher for oocytes that had been exposed to exogenous eGH versus controls (16/28, 57.1% versus 8/26, 30.8%, P < 0.05). In the present study, the BCB test was not useful for predicting competent equine oocytes prior to IVM. However, eGH enhanced equine oocyte maturation in vitro.

To understand sperm plug dissolution and spermatophore dehiscence in Portunidae, histological and ultrastructural changes in the seminal receptacle (SR) of Arenaeus cribrarius were investigated during ovarian development. In juvenile females the SR was filled with acid polysaccharides and the dorsal epithelium was stratified. Mated females with rudimentary ovaries showed a large SR filled by a glycoprotein sperm plug. This plug was present until the developing-ovary stage, when spermatophore dehiscence and intense holocrine secretions in the dorsal dense layer occurred. The plug was absent after the intermediate stage, and the SR became flaccid. The secretion produced moved the spermatophores into the ventral region. The modified dorsal epithelium in the transition between the dorsal and ventral regions released acid polysaccharides, which were found among the sperm, by exocytosis. The morphological changes of the SR in A. cribrarius, including the presence of the sperm plug, followed the macroscopic pattern observed in other members of Portunidae, such as blue crabs. However, in this species dissolution of the sperm plug was synchronized with ovarian development and occurred simultaneously with spermatophore dehiscence, showing the evolutionary relationship of the seminal receptacle and the female reproductive system to the storage of spermatophores and spermatozoa., (© 2014 Marine Biological Laboratory.)

This study was conducted to determine the age and liveweight at puberty of 120 crossbred beef heifers submitted to four diets to achieve predetermined weight gains (kg/day): 0.5 (G500; n = 32), 0.75 (G750; n = 32), 1.0 (G1000; n = 29), and 1.25 (G1250; n = 27). Animals were classified depending on their level of crossing between Nelore (N) and Hereford (H): 25%N-75%H, 37.5%N-63.5%H, 43.7%N-56.7%H, 50%N-50%H, and 75%N-25%H. Reproductive evaluation was performed at the beginning and at the end of the experimental period by ultrasonography and hormone analysis. The average age and liveweight at puberty were 388.0 ± 1.9 days and 331.4 ± 1.3 kg, respectively. Animals from the 25%N-75%H group reached puberty earlier than heifers from other genetic groups supplemented with G1250 diet (P < 0.05). Heifers with higher degree of Nelore (75%N-25%H) fed with G1000 diet showed estrus 42 days prior to mating, but only 57% reached puberty at mating (P < 0.05). Heifers with follicles of higher diameter reached puberty (P < 0.05) earlier. Higher average daily weight gain showed a positive effect on follicular diameter and IGF-I level at puberty (P < 0.01). Concentrations of GH were lower in heifers fed G1250 compared to G1000 diet (P < 0.05). There was a significant interaction between nutritional level and insulin levels at puberty (P < 0.01). We demonstrated the relationship between IGF-I and average daily gain on the onset of puberty in heifers. In conclusion, heifers submitted to the higher feeding level showed a higher follicular diameter and were younger at puberty.

Aging is associated with decreases in bone quality and in glomerular filtration. Consequently, osteoporosis and chronic kidney disease (CKD) are common comorbid conditions in the elderly, and often coexist. Biochemical abnormalities in the homeostasis of calcium and phosphorus begin early in CKD, leading to an increase in fracture risk and cardiovascular complications since early stages of the disease. The ability of DXA (dual energy X-ray absorptiometry) to diagnose osteoporosis and to predict fractures in this population remains unclear. The management of the disease is also controversial: calcium and vitamin D, although recommended, must be prescribed with caution, considering vascular calcification risk and the development of adynamic bone disease. Furthermore, safety and effectiveness of osteoporosis drugs are not established in patients with CKD. Thus, risks and benefits of antiosteoporosis treatment must be considered individually.

The Grb14 protein is a member of the Grb7 protein family. This protein family acts by binding to tyrosine kinase receptors, promoting cell proliferation and differentiation. There is evidence of the involvement of tyrosine kinase factors in the bovine oocyte maturation process. However, Grb14 has not been studied for bovine cumulus-oocyte complexes (COCs). The aim of the present study was to characterize Grb14 mRNA expression in bovine COCs during follicular development. Furthermore, we demonstrated that the expression of Grb14 mRNA is not regulated by estradiol. mRNA expression of Grb14 was assessed in 480 COCs from follicles of different sizes (1-3, 4-6, 6-8 or >8 mm) by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Grb14 mRNA expression decreased in COCs throughout follicular growth (P < 0.05). The role of estradiol in the expression of Grb14 mRNA in COCs was studied. Grb14 mRNA abundance did not differ in COCs cultured in the presence or absence of 17β-estradiol or fulvestrant. In conclusion, we showed that Grb14 mRNA is downregulated in COCs during antral follicle development, a finding that suggests a role for Grb14 in oocyte competence.

Background: In vitro fertilization (IVF) procedures are limited by the inability to mature equine oocytes on in vitro methods., Objective: The aim of this study was to evaluate structural integrity of equine oocytes subjected to vitrification with a synthetic polymer (PVA)., Methods: The effect of eGH and its relationship with IGF-I on in vitro maturation (IVM) were evaluated. Compact cumulus oocytes complexes (n=122) were cultured in TCM-199 with eGH, IGF-I or eGH+IGF-I for 30h at 38.5C in air with 5 % CO2. Oocytes were fixed after IVM or subjected to the vitrification protocol. Cryopreserved oocytes were exposed to 1.4M DMSO+1.8M EG+1 percent PVA for 3min, and then transfer to 2.8M DMSO+3.6M EG+0.6M sucrose+1 % PVA for 1min. After rewarming, oocytes were evaluated by confocal microscopy., Results: Maturation rates of oocytes were not significant different among groups (P > 0.05), however eGH+IGF-I group can develop the assessment of resumption of meiosis (MI+MII = 86.7 %)., Conclusion: The oocyte did not show morphological alterations. The use of PVA-copolymer may represent a potential alternative for vitrification of equine oocytes after IVM.

Twenty-seven 7-chloroquinolinotriazole derivatives with different substituents in the triazole moiety were synthesized via copper-catalyzed cycloaddition (CuAAC) click chemistry between 4-azido-7-chloroquinoline and several alkynes. All the synthetic compounds were evaluated for their in vitro activity against Plasmodium falciparum (W2) and cytotoxicity to Hep G2A16 cells. All the products disclosed low cytotoxicity (CC50 > 100 μM) and five of them have shown moderate antimalarial activity (IC50 from 9.6 to 40.9 μM). As chloroquine analogs it was expected that these compounds might inhibit the heme polymerization and SAR studies were performed aiming to explain their antimalarial profile. New structural variations can be designed on the basis of the results obtained., (Copyright © 2013 Elsevier Masson SAS. All rights reserved.)

Gonadal hormones influence physiological responses to stress and cutaneous wound healing. The aim of this study was to investigate the role of gonadal hormones on cutaneous wound healing in chronically stressed mice. Male and female mice were gonadectomized, and after 25 days, they were spun daily at 115 rpm for 15 min every hour until euthanasia. Twenty-eight days after the gonadectomy, an excisional lesion was created. The animals were killed 7 or 14 days after wounding, and the lesions were collected. Myofibroblast density, macrophage number, catecholamine level, collagen deposition, and blood vessel number were evaluated. In the intact and gonadectomized groups, stress increased the plasma catecholamine levels in both genders. In intact groups, stress impaired wound contraction and re-epithelialization and increased the macrophage number in males but not in females. In addition, stress compromised myofibroblastic differentiation and blood vessel formation and decreased collagen deposition in males but not in females. In contrast to intact mice, wound healing in ovariectomized female mice was affected by stress, while wound healing in castrated male mice was not. In conclusion, gender differences contribute to the cutaneous wound healing of chronically stressed mice. In addition, androgens contribute to the stress-induced impairment of the healing of cutaneous wounds but estrogens inhibit it., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Neosporosis has been considered the main cause of abortion between the first and the second trimester of pregnancy in cattle. Therefore, the objective of this study was to identify the presence of Neospora caninum DNA obtained from experimental models based on the evaluation of different areas of the fetal nervous system and organs from heifers previously inoculated with NC-1 after or before insemination. This study was performed with Hereford × Nelore (n=29) heifers and all animals were considered free of diseases at the beginning of the experiment. All animals were bred by fixed-time artificial insemination (TAI) and allocated as follows: (a) seronegative heifers subjected to TAI (TAI, n=9), (b) heifers infected with N. caninun 60 days prior to TAI (NC-1+TAI, n=9), and (c) heifers submitted to TAI and infected with N. caninum 60 days later (TAI+NC-1, n=11). The pregnancy was confirmed by transrectal ultrasonography 35 days after TAI and evaluated every 30 days until the end of gestation. Fetuses were collected surgically at 170 days of gestation, and immediately necropsied to remove tissues aseptically. Samples of the central nervous system (CNS), heart, kidney, lung, liver, skeletal muscle and caruncle were collected for DNA extraction. Days of gestation at abortion and interval from abortion to first insemination were examined by Student's t-test. At 35 days of gestation the pregnancy rates in the group NC-1+TAI (4/9, 44.4%) was lower than in the control group (8/9, 88.8%, P<0.05). At 60 days, the pregnancy rates in the NC-1+TAI group (0/4, 0%) was lower compared to TAI+NC-1 (5/7, 71.4%) and control (6/8, 75.0%) groups (P<0.05). Animals from the group NC-1+TAI were re-inseminated 60 days after the first TAI. After pregnancy losses throughout the study, 5 animals (TAI), 3 animals (NC-1+TAI) and 5 animals (TAI+NC-1) maintained pregnancy until 170 days of gestation. TaqMan RT-PCR demonstrated the presence of N. caninum DNA in the medulla and right posterior cortex in 3 out of 5 fetuses from the TAI+NC-1 group. We concluded that heifers infected after TAI had a higher incidence of the parasite at the fetus CNS. Identification of N. caninum by TaqMan RT-PCR would assist in the investigation of infection and in the evaluation of vaccines or therapeutic drugs to control neosporosis in cattle., (Copyright © 2013 Elsevier B.V. All rights reserved.)

The susceptibility of two fish and four mosquito species to the Caiman yacare haemoparasite Hepatozoon caimani was experimentally investigated. Mosquitoes belonging to four species (Aedes fluviatilis, Aedes albopictus, Aedes aegypti and Culex quinquefasciatus) were blood-fed on two naturally infected C. yacare from the Central-West Region of Brazil that exhibited distinct levels of parasitaemia: caimans A (11.05%) and B (1.25%). None of the engorged A. fluviatilis, A. albopictus or A. aegypti mosquitoes fed on caiman A survived for the duration of the sporogonic cycle; the great majority of the engorged mosquitoes died within 48 h of the blood meal. All A. aegypti fed on caiman B were negative, whereas 91.3% of dissected C. quinquefasciatus fed on the same caiman contained oocysts. Characid fish-Metynnis sp. and Astyanax sp.-were individually fed with C. quinquefasciatus females previously engorged (21-23 days) on caiman B. No parasite was found in the Astyanax fish. By contrast, 100% of the Metynnis fish depicted numerous cysts harbouring cystozoites identical to those of H. caimani, even more than 8 months after the ingestion of the infected mosquitoes. The cysts were located near the veins of the liver and, in some cases, close to the tunica intima of these vessels. No inflammatory reaction was observed. Gametocytes were observed in the blood smears of juvenile caimans that had ingested infected fish 9-12 weeks earlier. The potential role of fish as paratenic vertebrate hosts of H. caimani in nature is discussed.

Chlorthalidone (CTD) is an antihypertensive drug and exhibits four crystalline forms: I, II, III and IV. In this paper, the incidence of CTD polymorphs in raw materials and in tablets as well as the solubility and dissolution properties of forms I and II have been studied. Raw materials were named as A, B, C, D, and E and tablets as Reference, G1, G2 and S. Using powder X-ray diffraction and infrared spectroscopy analyses we found that A, B, E, Reference and G1 contain CTD form I; C, D and S contain predominantly form II; and G2 contain a mixture of both forms. Solubility experiments showed that form II is up to 49% more soluble than form I and dissolution studies showed a significantly effect of the polymorphism on the dissolution of CTD from tablets. Based on these results, it was concluded that only the CTD form I is acceptable for preparation of tablet form. Moreover, we proposed the polymorphic quality control of CTD raw materials and tablets., (Copyright © 2013 Elsevier B.V. All rights reserved.)

The objectives of this study were to evaluate the effects of equine growth hormone (eGH) on nuclear and cytoplasmic maturation of equine oocytes in vitro, steroid production by cumulus cells, and expression and subcellular localization of eGH-receptors (eGH-R) on equine ovarian follicles. Cumulus-oocyte complexes (COCs) were recovered by aspirating follicles <30 mm in diameter from abattoir-derived ovaries. The COCs were morphologically evaluated and randomly allocated to be cultured in either a control maturation medium or supplemented with 400 ng/mL eGH, for 30 h at 38.5°C in air with 5% CO2. The COCs were stained with 10 μg/mL propidium iodide and 10 μg/mL fluorescein isothiocyanate-labeled Lens culinaris agglutinin. Chromatin configuration and distribution of cortical granules were assessed via confocal microscopy. Compared to control, COCs incubated with eGH had: more oocytes that reached metaphase II (35/72, 48.6% vs. 60/89, 67.4%, respectively; P=0.02); greater concentrations of testosterone (0.21 ± 0.04 vs. 0.06 ± 0.01 ng/mL; P=0.01), progesterone (0.05 ± 0.01 vs. 0.02 ± 0.00 ng/mL; P=0.04), and oestradiol (76.80 ± 14.26 vs. 39.58 ± 8.87 pg/mL; P=0.05) in the culture medium, but no significant differences in concentration of androstenedione. Based on Real Time RT-PCR analyses, expression of the eGH-R gene was greater in cumulus cells and COCs at the start than at the end of in vitro maturation. Positive immunostaining for eGH-R was present in cumulus cells, the oocytes and granulosa cells. In conclusion, addition of eGH to maturation medium increased rates of cytoplasmic maturation and had an important role in equine oocyte maturation, perhaps mediated by the presence of eGH-R in ovarian follicles., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

In horses, successful in vitro fertilization procedures are limited by our inability to consistently mature equine oocytes by in vitro methods. Growth hormone (GH) is an important regulator of female reproduction in mammals, playing an important role in ovarian function, follicular growth and steroidogenesis. The objectives of this research were to investigate: the effects of equine growth hormone (eGH) and insulin-like growth factor-I (IGF-I) on the in vitro maturation (IVM) of equine oocytes, and the effects of eGH in addition to estradiol (E2), gonadotropins (FSH and LH) and fetal calf serum (FCS) on IVM. We also evaluated the cytoskeleton organization of equine oocytes after IVM with eGH. Equine oocytes were aspirated from follicles <30 mm in diameter and matured for 30 h at 38.5°C in air with 5% CO2. In experiment 1, selected cumulus-oocyte complexes (COCs) were randomly allocated as follows: (a) control (no additives); (b) 400 ng/ml eGH; (c) 200 ng/ml IGF-I; (d) eGH + IGF-I; and (e) eGH + IGF-I + 200 ng/ml anti-IGF-I. In addition to these treatment groups, we also added 1 μg/ml E2, 5 IU/ml FSH, 10 IU/ml LH and 10% FCS in vitro (experiment 2). Oocytes were stained with markers for microtubules (anti-α-tubulin antibody), microfilaments (AlexaFluor 488 Phalloidin) and chromatin (TO-PRO3-iodide) and assessed via confocal microscopy. No difference was observed when eGH and IGF-I was added into our IVM system. However, following incubation with eGH alone (40%) and eGH, E2, gonadotropins and FCS (36.6%) oocytes were classified as mature v. 17.6% of oocytes in the control group (P < 0.05). Matured equine oocytes showed that a thin network of filaments concentrated within the oocyte cortex and microtubules at the metaphase spindle showed a symmetrical barrel-shaped structure, with chromosomes aligned along its midline. We conclude that the use of E2, gonadotropins and FCS in the presence of eGH increases the number of oocytes reaching oocyte competence.

This study aims at investigating the effect of removing the negative values of Choi-Wiliams distribution (CWD) related to the electromyogram (EMG) for visualization and instantaneous median frequency (IMF) estimation. Beyond the EMG signals from triceps surae and biceps brachialis, the CWD was applied in a simulated sinusoidal signal as like in stationary and non-stationary simulated EMG signals (SES). The CWD negative values of all simulated and EMG signals were removed. The IMF values were obtained for SES and EMG. The CWD IMF values from SES and EMG were thus compared with the IMF values from short time Fourier transform (STFT) by means of correlation. The suppression of negative values from the CWD reduced cross terms influence and improved visualization, as shown by the increased correlation coefficient between the IMF values. Before this suppression, the extracted IMF values showed large oscillation along the time, with various spurious values beyond 500Hz, which disappeared after the suppression. Moreover, this procedure seems to be especially useful for non-stationary signals., (Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.)

Objective: To examine the influence of a preventative training program (PTP) on sagittal plane kinematics during different landing tasks and vertical jump height (VJH) in males., Design: Six weeks prospective exercise intervention., Participants: Fifteen male volleyball athletes (13 ± 0.7 years, 1.70 ± 0.12 m, 60 ± 12 kg)., Interventions: PTP consisting of plyometric, balance and core stability exercises three times per week for six weeks. Bilateral vertical jumps with double leg (DL) and single leg (SL) landings were performed to measure the effects of training., Main Outcome Measurements: Kinematics of the knee and hip before and after training and VJH attained during both tasks after training. The hypothesis was that the PTP would produce improvements in VJH, but would not generate great changes in biomechanical behavior., Results: The only change identified for the SL was the longest duration of landing, which represents the time spent from initial ground contact to maximum knee flexion, after training, while increased angular displacement of the knee was observed during DL. The training did not significantly alter the VJH in either the SL (difference: 2.7 cm) or the DL conditions (difference: 3.5 cm)., Conclusions: Despite the PTP's effectiveness in inducing some changes in kinematics, the changes were specific for each task, which highlights the importance of the specificity and individuality in selecting prevention injury exercises. Despite the absence of significant increases in the VJH, the absolute differences after training showed increases corroborating with the findings of statistically powerful studies that compared the results with control groups. The results suggest that short-term PTPs in low risk young male volleyball athletes may enhance performance and induce changes in some kinematic parameters., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

The objective of this study was to test the hypothesis that equine growth hormone (eGH), in combination with insulin growth factor-I (IGF-I), influences positively in vitro nuclear and cytoplasmic maturation of equine oocytes. Cumulus-oocyte complexes were recovered from follicles that were < 25 mm in diameter, characterized by morphology and were allocated randomly as follow: (a) control (no additives); (b) 400 ng/ml eGH; (c) 200 ng/ml IGF-I; (d) eGH + IGF-I; and (e) eGH + IGF-I + 400 ng/ml anti-IGF-I antibody. Oocytes were matured for 30 h at 38.5°C in air with 5% CO2 and then stained with 10 μg/ml propidium iodide (PI) to evaluate nuclear status and 10 μg/ml Lens culinaris agglutinin-fluorescein complex (FITC-LCA) to assess cortical granule migration by confocal microscopy. The proportion of immature oocytes that developed to the metaphase II (MII) stage in the eGH + IGF-I group (15 of 45) was greater than in the groups that were treated only with IGF-I (7 of 36, p = 0.03). Oocytes that reached MII in the control group (20 of 56; 35.7%) showed a tendency to be different when compared with eGH + IGF-I group (15 of 45; 33.3%, p = 0.08). The treated group that contained anti-IGF-I (15 of 33; 45.4%) decreased the number of oocytes reaching any stage of development when compared with eGH (47 of 72; 65.3%) and eGH + IGF-I (33 of 45; 73.3%) groups (p = 0.05) when data from MI and MII were combined. We concluded that the addition of eGH to in vitro maturation (IVM) medium influenced the in vitro nuclear and cytoplasmic maturation of equine oocytes. The use of GH and IGF-I in vitro may represent a potential alternative for IVM of equine oocytes.

An X-ray transmission microtomography (CT) system combined with an X-ray fluorescence microtomography (XRFμCT) system was implemented in the Brazilian Synchrotron Light Laboratory (LNLS), in order to determine the elemental distribution in prostate samples aiming at establishing a correlation between the concentration of some elements and the characteristics and pathology of the tissues. The CT images were reconstructed using a filtered-back projection algorithm and the XRFμCT images were reconstructed using a filtered-back projection algorithm with absorption corrections., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The aim of this study was to evaluate the effect of leukemia inhibitory factor (LIF) on the activation and survival of preantral follicles cultured in vitro enclosed in ovarian fragments (in situ). Goat ovarian cortex was divided into fragments to be used in this study. One fragment was immediately fixed (fresh control - FC) and the remaining fragments were cultured in supplemented minimum essential medium (MEM) without (cultured control - CC) or with different concentrations of LIF (1, 10, 50, 100 or 200 ng/ml) for 1 or 7 days, at 39°C in air with 5% CO2. Fresh control, CC and treated ovarian fragments were processed for histological and fluorescence analysis. The percentage of histological normal preantral follicles cultured for 7 days with 1 ng/ml (49.3%), 10 ng/ml (58.6%) and 50 ng/ml (58%) of LIF was higher than in the CC (32.6%; p < 0.05). After 7 days of culture, the percentage of primordial follicles in situ cultured with LIF decreased and primary follicles increased in all LIF concentrations compared with FC and CC (p < 0.05). In conclusion, LIF induced primordial follicle activation and supported preantral follicle viability of goat ovarian tissues cultured for 7 days.