Search

Tumors can exert a far-reaching influence on the body, triggering systemic responses that contribute to debilitating conditions like cancer cachexia. To characterize the mechanisms underlying tumor-host interactions, we utilized a BioID-based proximity labeling method to identify proteins secreted by Yki act adult Drosophila gut tumors into the bloodstream/hemolymph. Among the major proteins identified are coagulation and immune-responsive factors that contribute to the systemic wasting phenotypes associated with Yki act tumors. The effect of innate immunity factors is mediated by NFκB transcription factors Relish, dorsal, and Dif, which in turn upregulate the expression of the cachectic factors Pvf1, Impl2, and Upd3. In addition, Yki act tumors secrete Eiger, a TNF-alpha homolog, which activates the JNK signaling pathway in neighboring non-tumor cells, leading to cell death. The release of damage-associated molecular patterns (DAMPs) from these dying cells presumably amplifies the inflammatory response, exacerbating systemic wasting. Targeting the inflammatory response, the JNK pathway, or the production of cachectic factors could potentially alleviate the debilitating effects of cancer cachexia., Competing Interests: Conflicts of Interest. The authors declare no conflict of interest.

This study examined the effect of a competitive season on salivary responses [cortisol (sC), testosterone (sT), Testosterone/Cortisol ratio (sT/C), Immunoglobulin A (sIgA), sIgA secretion rate (srIgA), alpha-amylase (sAA)] and upper respiratory symptoms (URS) occurrence in three teams of male soccer players (Under-15, Under-17 and Under-19 yrs.). Training and competition volumes, salivary biomarkers and URS were determined monthly. No differences were found for monthly training volume between teams. Incidence of URS was higher for the U15 (44.9% of the total cases). Higher sT and srIgA were observed for the U19, lower sC were found for the U17 and sAA showed higher values for the U15 throughout the season. In the U15, significant difference (p = .023) was found for sIgA concentration with higher concentration values in January compared to December (-42.7%; p = .008) and the sT showed seasonal variation (p < .001) with the highest value in January significantly different from October (-40.2%; p = .035), November (-38.5%; p = 0.022) and December (-51.6%; p = .008). The U19 presented an increase in sC in March compared to February (-66.1%, p = .018), sT/C were higher in February compared to March (-58.1%; p = .022) and sAA increased in March compared to September (-20.5%; p = .037). Negative correlations, controlled for age group, were found between URS occurrence and srIgA (r = -0.170, p = .001), sAA (r = -0.179, p = .001) and sT (r = -0.107, p = .047). Monitoring salivary biomarkers provides information on mucosal immunity with impact in URS occurrence. Coaches could manipulate training loads to attenuate the physical stressors imposed on athletes, especially at demanding and stressful periods., Competing Interests: The authors have declared that no competing interests exist.

Vitamin D supplementation is universal for postmenopausal women, but not for elderly men, in whom osteoporosis is also commonly neglected. This study aimed to evaluate vitamin D deficiency and its association with secondary hyperparathyroidism, bone resorption, and bone density in Brazilian men. A total of 120 men, 20-93 years, were evaluated for serum calcium, phosphorus, creatinine, 25-hydroxyvitamin D (25(OH)D), parathyroid hormone, biochemical markers of bone resorption (carboxy-terminal telopeptide, carboxy-terminal peptide of type I collagen), and bone mineral density (dual-energy X-ray absorptiometry). Glomerular filtration rate (GFR) below 30 mL/min/1.73 m 2 , chronic diseases, and medications affecting bone were the exclusion criteria. No participant reported previous low-impact fractures. In the overall population, 25(OH)D levels were below 30 ng/mL in 46.7%, and below 20 ng/mL in 27.6%. Among the 93 patients 50 years and older, 28 had osteoporosis. In those 70 years and older, the prevalence of vitamin D deficiency (42.1%), secondary hyperparathyroidism (46.4%), high bone resorption (39.6%), decreased GFR (39.2%), and osteoporosis (41.4%) was significantly higher than in the younger subjects (p < 0.005 for all comparisons). Serum parathyroid hormone increased with aging and declining GFR, but was not significantly associated with 25(OH)D or bone mineral density. There was a clear contribution of vitamin D deficiency to increased bone resorption and osteoporosis. Binary logistic regression model considering age, 25(OH)D, and bone resorption identified age ≥70 years as the main determinant of osteoporosis. Our data demonstrate a high prevalence of vitamin D deficiency in a male population living in Rio de Janeiro, and emphasize its participation on the pathogenesis of age-related bone loss. (Vitamin D deficiency and osteoporosis are common in elderly Brazilian men.)., (Copyright © 2017 The International Society for Clinical Densitometry. Published by Elsevier Inc. All rights reserved.)

Context aware systems are able to adapt their behavior according to the environment in which the user is. They can be integrated into an Internet of Things (IoT) infrastructure, allowing a better perception of the user's physical environment by collecting context data from sensors embedded in devices known as smart objects. An IoT extension called the Internet of Mobile Things (IoMT) suggests new scenarios in which smart objects and IoT gateways can move autonomously or be moved easily. In a comprehensive view, Quality of Context (QoC) is a term that can express quality requirements of context aware applications. These requirements can be those related to the quality of information provided by the sensors (e.g., accuracy, resolution, age, validity time) or those referring to the quality of the data distribution service (e.g, reliability, delay, delivery time). Some functionalities of context aware applications and/or decision-making processes of these applications and their users depend on the level of quality of context available, which tend to vary over time for various reasons. Reviewing the literature, it is possible to verify that the quality of context support provided by IoT-oriented middleware systems still has limitations in relation to at least four relevant aspects: (i) quality of context provisioning; (ii) quality of context monitoring; (iii) support for heterogeneous device and technology management; (iv) support for reliable data delivery in mobility scenarios. This paper presents two main contributions: (i) a state-of-the-art survey specifically aimed at analyzing the middleware with quality of context support and; (ii) a new middleware with comprehensive quality of context support for Internet of Things Applications. The proposed middleware was evaluated and the results are presented and discussed in this article, which also shows a case study involving the development of a mobile remote patient monitoring application that was developed using the proposed middleware. This case study highlights how middleware components were used to meet the quality of context requirements of the application. In addition, the proposed middleware was compared to other solutions in the literature., Competing Interests: The authors declare no conflict of interest.

The aim of this study was to assess in vitro meiosis resumption and nuclear maturation of Rattus norvegicus oocytes after vitrification with different cryoprotective solutions. Cumulus-oocyte complexes (COCs) were exposed to an equilibration solution for 4 min placed in cryoprotective solutions for 1 min and vitrified in open pulled straws. Cryoprotective solutions were prepared with 15% ethylene glycol + 15% dimethyl sulfoxide + 0.5 M sucrose and different supplements, to form the following groups: G1, 20% fetal bovine serum in modified phosphate-buffered saline (mPBS); G2, 0.4% bovine serum albumine in mPBS; G3, 1% hyaluronic acid in mPBS; and G4, 0.4% polyvinyl alcohol in mPBS. Seven days after vitrification, the COCs from G1 to G4 were warmed and in vitro matured for 30 h along with the control group. Hoechst staining was performed to assess meiosis resumption and nuclear maturation rates. Control group showed higher meiosis resumption (77.88%) and nuclear maturation rates (55.75%) compared to all vitrified groups. Among the vitrified COCs, G3 showed the highest meiosis resumption and nuclear maturation rates (G1, 26.5 and 15.38%; G2, 22.12 and 11.54%; G3, 34.55 and 20%; G4, 20.17 and 9.24%). Supplementation of the vitrification solution with 1% hyaluronic acid provided better results, compared to the other supplements. Hyaluronic acid can be useful to vitrify rat COCs associated with other cryoprotectant agents.

Oocyte selection based on the brilliant cresyl blue (BCB) staining test has been successfully used to differentiate between competent and incompetent bovine oocytes. Here, the expression of genes involved in transport of monocarboxylates (Mct1-4) and oogenesis specific genes (Bmp15, Gdf9 and Has2) in BCB+ and BCB- selected immature and mature bovine cumulus-oocyte complexes (COC) was evaluated. In order to find specific molecular markers to characterize successful oocyte maturation, our study was also aimed at identifying the expression of Mcts and oogenesis specific genes in denuded oocytes and cumulus cells. Immature COCs morphological appropriate were (i) stained with 26 mm BCB for 90 min before IVM, (ii) exposed to same incubation conditions as stained COCs, but without BCB (holding group) or (iii) transferred into a maturation medium immediately after morphological selection (control group). mRNA expression was investigated by RT-PCR in COCs before and after IVM. No relationship was observed in the relative expression of Has2, Gdf9, Bmp15 or Mct1, 2 and 4 transcripts between BCB+ and BCB- COCs. Transcripts analysis showed that Gdf9 and Bmp15 in BCB+, BCB- and holding groups were up-regulated (p < 0.05) before IVM, while Has2 was up-regulated (p < 0.01) after IVM in the control group. Other genes remained stable during maturation (Mct1, 2 and 4). Our results showed, for the first time, Mct1, 2 and 4 expression in bovine COCs. Mct1 and Mct4 transcripts were present in denuded oocytes and cumulus cell, while Mct2 was detected only in cumulus cells. These differences between the three isoforms in localization suggest unique roles for each in monocarboxylate transport during maturation., (© 2015 Blackwell Verlag GmbH.)

Objective: To establish the injury profile of soccer players from a first division Brazilian soccer team. In addition, we investigated the association between the characteristics of the injuries and the player's age and position., Method: Forty-eight players from a Brazilian first division soccer team were followed during one season. Descriptive statistics were used to characterize the injury profile. Spearman's tests were used to verify the association between the number and severity of injuries and the player's age. Chi-square test was used to verify the association between type of injury and player's position. Fisher's exact test was used to verify the association between the severity of injuries and player's position., Results: The incidence of injuries was 42.84/1000 hours in matches and 2.40/1000 hours in training. The injury severity was 19.5±34.4 days off competition or training. Lower limb was the most common location of injury and most injuries were muscular/tendinous, overuse, non-recurrent, and non-contact injuries. Player's age correlated with the amount and severity of muscle and tendon injuries. Defenders had more minimal injuries (1-3 days lost), while forwards had more moderate (8-28 days lost) and severe injuries (>28 days lost). Furthermore, wingbacks had more muscle and tendon injuries, while midfielders had more joint and ligament injuries., Conclusion: The injury profile of the Brazilian players investigated in this study reflected regional differences in soccer practices. Results confirm the influence of the player's age and position on the soccer injuries profile.

Objectives/hypothesis: The aim of this study has been to establish an alternative approach in the form of regeneration of the thyroid cartilage., Study Design: Four 1-month old pigs (Sus scrofa) were used (divided into 3 groups) and submitted to general anesthetic to perform cervictomy with exposure of the thyroid cartilage in a total of 12 (twelve) samples., Method: A resection of 4.0 cm(2) of cartilage was carried out in the right upper region and in the left upper and lower left region of the cartilage, where a scaffold with or without stem cells was implanted. In the left lower region, no biomaterial was implanted and the defect was left open (lesion control [L])., Results: The average extension of the cartilaginous neoformation of L group was 136.3 μm (± 9.6) and 387.7 μm (± 43.2) in the scaffold (SCA) group, presenting a significant statistical difference (P < 0.01). The analysis carried out on the lesion site sections of the cartilage of the larynx of the animals from the SCA group + mesenchymal stem cells (SCA+MSC) showed an average of the extension of neocartilage of 825.4 μm (± 122.1), showing a more extensive area of neocartilage when compared to the other groups. These results demonstrated a high significantly statistical difference (P < 0.001) when compared with the L and SCA groups., Conclusion: In 100% of the cases for which SCA+MSCs were used, a significant success in the cartilage growth and closing of the lesion in the thyroid cartilage was obtained compared to the other two groups for which MSCs were not used., Level of Evidence: N/A., (© 2014 The American Laryngological, Rhinological and Otological Society, Inc.)

A time discrete age-structured model for modeling the spread of Dengue fever is built. The demographic dynamics is introduced trough the Leslie model. The basic reproductive number is introduced, and an approximation for it is built. The final age distributions for the susceptibles, infected and removed are obtained, and we show how they can be used to produce an actual estimate for R0 from stratified serological data. An application is made using data from Recife, Brazil, and explicit estimates for R0 are given.

The objective of this work was to evaluate the rate of meiosis resumption and nuclear maturation of rat (Rattus norvegicus) oocytes selected for in vitro maturation (IVM) after staining of cumulus-oocyte complexes (COCs) with blue cresyl brilliant (BCB) using different protocols: exposure for 30, 60 or 90 min at 26 μM BCB (Experiment 1), and exposure for 60 min at 13, 20 or 26 μM BCB (Experiment 2). In Experiment 1, the selection of oocytes exposed to BCB for 60 min was found to be the most suitable, as meiosis resumption rates in the BCB(+) group (n = 35/61; 57.37%) were the closest to the observed in the control (not exposed) group (n = 70/90; 77.77%) and statistically higher than the values observed for the BCB(-) group (n = 3/41; 7.32%). Additionally, the more effective evaluation of diagnostic tests (sensitivity and negative predictive value 100%) was observed in COCs exposed for 60 min. In Experiment 2, the 13 μM BCB(+) group presented rates of meiosis resumption (n = 57/72; 72.22%) similar to the control group (n = 87/105; 82.86%) and higher than other concentration groups. However, this results of the analysis between BCB(-) oocytes was also higher in the 13 μM BCB group (n = 28/91; 30.78%) when compared with BCB(-) COCs exposed to 20 μM (n = 3/62; 4.84%) or 26 μM (n = 3/61; 4.92%) BCB. The nuclear maturation rate in the 13 μM BCB group was similar between BCB(+) or BCB(-) oocytes. The 20 μM BCB group had a lower rate of nuclear maturation of BCB(-) oocytes than other groups. Thus, our best results in the selection of Rattus norvegicus oocytes by staining with BCB were obtained using the concentration of 13 μM and 20 μM, and an incubation period of 60 min.

Background: Persistent asthma in children is a chronic inflammatory disease and glucocorticoids (GCs) are currently recognized as the mainstay of therapy. Clinical and in vitro steroid resistance has been demonstrated in severe asthma. However, GC insensitivity has not been studied in children with controlled persistent asthma., Objectives: To analyze peripheral blood mononuclear cell (PBMC) sensitivity to GC in children (6-15 years) with persistent asthma and healthy controls., Methods: Children with persistent asthma were selected and lung function and skin-prick tests were performed in all studied asthmatic children. PBMCs were isolated and cultured in vitro to assess mitogen-induced proliferation and cellular sensitivity to dexamethasone., Results: Fifty-seven children with persistent and controlled asthma (mean age 10 years) were recruited and divided into 3 groups (severe, moderate and mild), and compared to healthy children (n = 18). Children with asthma, regardless of the severity of disease, presented similar sensitivity to GCs when compared to healthy children. Patients with mild asthma showed significantly less sensitivity to dexamethasone and children with severe asthma had similar sensitivity to dexamethasone when compared to controls., Conclusions: In vitro insensitivity to GCs was not demonstrated in children with controlled persistent asthma, even in those with severe disease. Our findings suggest that resistance to GCs in older patients with severe asthma might be an acquired process. However, future longitudinal studies are necessary to confirm this hypothesis., (Copyright © 2010 S. Karger AG, Basel.)

Summary: Osteoporosis in men is underestimated, but our data point to an increasing prevalence rate in those over 70 years old with body mass index (BMI) <25 kg/m(2), bioavailable testosterone <2.7 nmol/L, bioavailable estradiol <40 pmol/L, and high bone turnover, defined in this study as serum carboxyterminal cross-linked telopeptide of type I collagen (ICTP) >4.3 microg/L., Introduction: The association of sex steroids and osteoporosis was evaluated in 104 men, aged 50-93 years old., Methods: Bone mineral density (BMD), bone turnover (ICTP), testosterone (T), and estradiol (E(2)) were measured; free and bioavailable hormones (free testosterone index [FTI], BioT, free estradiol index [FEI], and BioE(2)) were calculated from T, E(2), sex hormone-binding globulin (SHBG), and albumin. Nonparametric analysis and Poisson regression models were used., Results: Significant increases in SHBG and ICTP and decreases in femoral neck BMD, FTI, FEI, BioT, and BioE(2) were observed with each additional decade of age. Femoral neck BMD was inversely correlated with ICTP, and both were significantly associated with SHBG, FTI, BioT, FEI, and BioE. There was a direct and graded association between age and osteoporosis prevalence rate (OP PR; p = 0.028). Compared to participants less than 70 years old, the crude OP PR of those 80 years and older was 3.2 (95%CI = 1.4-7.3). Adjusting sequentially for BMI and bioavailable sex hormones attenuated the association between age and osteoporosis prevalence by 55% and 77%, respectively., Conclusion: Our data support the view that low BMI and declining sex steroids explain most of the association between aging, increased bone turnover, and osteoporosis in men.

In order to study the benefit of adding recombinant human growth hormone (rhGH) to antiresorptive therapy, six patients with idiopathic osteoporosis (IO) receiving alendronate plus calcium and vitamin D were started on daily subcutaneous injections of rhGH 2.0 IU for one year. Fasting morning urine and serum samples were collected for N telopeptide of type-1 collagen (NTX), serum bone-specific alkaline phosphatase (BSAP) and insulin-like growth factor 1 (IGF-1) during the study. Bone mineral density (BMD) was determined by dual-energy x-ray absorptiometry at baseline and 01 year. The effect of rhGH was evaluated comparing the percentage changes in BMD during the last year on ALN with the results obtained with the combined therapy. Serum IGF-1 increased in all patients but variations were not significant (p=0.266). Serum BSAP did not significantly change (p=0.078) but median NTX increased at 45 days from 12.3 to 19.8 nMBCE/mMCr (p=0.012) and tended to return to baseline values at 12 months (15.2 nMBCE/mMCr). Comparing with isolated ALN therapy, a beneficial effect on bone density was observed in 2/3 of the patients at lumbar spine, and percentage change (median and quartiles) varied from -0.65% (-2.33 and 2.23) on ALN to 0.70% (-0.35 and 3.03) on ALN+GH. Although no bone gain occurred at the femoral neck, our data point to a positive effect of rhGH in patients with idiopathic osteoporosis.

The objective of this study was to determine the developmental rates and relative abundance of Hsp 70.1 and Glut-1 transcripts in in vivo- and in vitro-produced (IVP) bovine embryos in media supplemented with bovine serum albumin (BSA) or different oestrous cow serum concentrations. In experiment 1, in vitro maturation and culture media were supplemented with 0.4% BSA or 1, 5, 10 or 20% of oestrous cow serum (ECS). The analysis of the expression of Hsp 70.1 and Glut-1 was carried out in individual days 7 and 8 embryos by a semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) assay. In experiment 2, in vivo-produced morulae were collected on day 7 of the oestrous cycle and employed for the comparison of the relative abundances of Hsp 70.1 and Glut-1 transcripts with IVP morulae produced using two protein sources (10% ECS or 0.4% BSA). No differences were observed in cleavage rate among groups, but blastocyst formation (27%) and hatching rates (78%) were significantly higher in IVP embryos produced with 20% ECS than the other groups (p<0.05). No significant differences were observed in the relative abundances of Hsp 70.1 and Glut-1 mRNA in days 7 and 8 blastocysts expanded blastocysts between groups. The abundances of mRNA for those genes were similar between IVP and in vivo-produced morulae. In spite of the alterations observed in embryonic development, the presence of serum at distinct concentrations did not appear to alter the relative abundance profiles of Hsp 70.1 and Glut-1 compared with controls or the BSA supplementation to the IVP media.

The objective of this study was to determine the effects of oocyte/embryo density (embryo number:medium volume ratio) on developmental competence and relative transcription of developmentally important genes in in vitro-produced bovine embryos. Using standard procedures, bovine oocytes were matured, fertilized and cultured in vitro in groups of 5, 10, 20 or 30 oocytes/embryos per 100 microL medium drop (embryo number/medium volume ratios of 1:20, 1:10, 1:5 and 1:3.3, respectively). A semi-quantitative RT-PCR assay was used to determine the relative abundance of Hsp 70.1 and Glut-1 transcripts in Day-6 morulae, and in Day-7 and Day-8 blastocysts and expanded blastocysts. Cleavage rates (55.9, 49.1, 47.1 and 34.6%, respectively) and blastocyst developmental rates (24.1, 18.9, 20.4 and 12.6%, respectively) were higher in groups containing 5, 10 and 20 embryos versus 30 embryos per drop, but there were no significant differences among groups in hatching rates. The relative abundance of Hsp 70.1 transcripts in Day-7 blastocysts was higher in embryos produced in the 20-embryo than in the 5-embryo groups. There were no differences among embryos from all groups in the amount of Glut-1 mRNA (regardless of embryonic stage). However, there were differences in the pattern of Hsp 70.1 gene expression between distinct stages of embryo development and embryo density; these dissimilarities appeared to be associated with compromised development at higher embryo densities. In conclusion, the increase in embryo density during IVM, IVF, and IVC phases of in vitro embryo production might negatively affect embryo development and the pattern of gene expression of in vitro-produced bovine embryos.

Improvements on embryo micromanipulation techniques led to the use of embryo bisection technology in commercial embryo transfer programs, and made possible the direct genetic analysis of preimplantation bovine embryos by biopsy. For example, aspiration and microsection, allow bovine embryos sexing by detection of male-specific Y-chromosome in a sample of embryonic cells. We report on the application of the methodologies of splitting and biopsy of bovine embryos in field conditions, and on the results of embryo sex determination by the polymerase chain reaction (PCR). Pregnancy rates achieved with fresh bisected or biopsied embryos (50 to 60%) were similar to the fresh intact embryos (55 to 61%). The PCR protocol used for embryo sexing showed 92% to 94% of efficiency and 90 to 100% of accuracy. These results demonstrate these procedures are suitable for use in field conditions.

The aim of this study was to develop a polymerase chain reaction (PCR) protocol for the detection of Salmonella in artificially contaminated chicken meat. Tests were performed with different dilutions of Salmonella Typhimurium or Salmonella Enteritidis cells (10(-7), 10(-8) or 10(-9) CFU/mL) inoculated in chicken meat samples, in order to establish the limits of detection, incubation times (0, 6, 8 and 24 hours of pre-enrichment in PBW 1%) and three DNA extraction protocols (phenol-chloroform, thermal treatment and thermal treatment and Sephaglass). The assay was able to detect until 10(-9) CFU/mL of initial dilution of Salmonella cells inoculated in chicken meat, which allows detection of Salmonella within 48 hours, including 24 hours of pre-enrichment and using the phenol-chloroform DNA extraction protocol. As the results are obtained in a shorter time period than that of microbiological culture, this procedure will be useful in the methodology for detection of Salmonella in chicken.