WHOLE GENOME TARGETED ENRICHMENT AND SEQUENCING OF HUMAN-INFECTING CRYPTOSPORIDIUM spp.

Cryptosporidium spp. are protozoan parasites that cause severe illness in vulnerable human populations. Obtaining pure Cryptosporidium DNA from clinical and environmental samples is challenging because the oocysts shed in contaminated feces are limited in quantity, difficult to purify efficiently, may derive from multiple species, and yield limited DNA (<40 fg/oocyst). Here, we develop and validate a set of 100,000 RNA baits (CryptoCap_100k) based on six human-infecting Cryptosporidium spp. (C. cuniculus, C. hominis, C. meleagridis, C. parvum, C. tyzzeri, and C. viatorum) to enrich Cryptosporidium spp. DNA from a wide array of samples. We demonstrate that CryptoCap_100k increases the percentage of reads mapping to target Cryptosporidium references in a wide variety of scenarios, increasing the depth and breadth of genome coverage, facilitating increased accuracy of detecting and analyzing species within a given sample, while simultaneously decreasing costs, thereby opening new opportunities to understand the complex biology of these important pathogens.

WHOLE GENOME TARGETED ENRICHMENT AND SEQUENCING OF HUMAN-INFECTING CRYPTOSPORIDIUM spp.

Cryptosporidium spp. are protozoan parasites that cause severe illness in vulnerable human populations. Obtaining pure Cryptosporidium DNA from clinical and environmental samples is challenging because the oocysts shed in contaminated feces are limited in quantity, difficult to purify efficiently, may derive from multiple species, and yield limited DNA (<40 fg/oocyst). Here, we develop and validate a set of 100,000 RNA baits (CryptoCap_100k) based on six human-infecting Cryptosporidium spp. ( C. cuniculus , C. hominis , C. meleagridis , C. parvum , C. tyzzeri , and C. viatorum ) to enrich Cryptosporidium spp. DNA from a wide array of samples. We demonstrate that CryptoCap_100k increases the percentage of reads mapping to target Cryptosporidium references in a wide variety of scenarios, increasing the depth and breadth of genome coverage, facilitating increased accuracy of detecting and analyzing species within a given sample, while simultaneously decreasing costs, thereby opening new opportunities to understand the complex biology of these important pathogens.

Use of ammonium chloride to prevent urolithiasis in sheep / [Utilização do cloreto de amônio na prevenção de urolitíase em ovinos]

Urolithiasis has a high incidence among confined sheep. It is multifactorial and may cause economic damage. Our aim was to determine the capacity of urinary acidification using ammonium chloride in sheep. Twenty-five 3-month-old male sheep were confined and randomly divided into three groups; the G200 and G500 groups received 200mg/kg/GW and 500mg/kg/GW of ammonium chloride daily for 56 consecutive days, respectively, whereas the CG group did not receive ammonium chloride. Sampling times and clinical evaluation were performed weekly, starting from the 14th day of confinement (M1 or immediately before administering ammonium chloride) until the 17th day (M9) of the feedlot. Hemogasometry, biochemical examination of serum urea and creatinine concentration and ultrasound evaluation of the urinary tract were performed. The urinalysis indicated a higher incidence of ammonium magnesium phosphate crystals at the beginning of the study, showing a migration to urate crystal formation, mainly in the G500 group because of urinary acidification. No hemogasometric, serum biochemistry, ruminal fluid, or ultrasonographic changes were observed. Urinary acidification was achieved and maintained after M7 during the administration of ammonium chloride in the G500 group, but not in the other study groups.(AU)

A urolitíase apresenta alta incidência em ovinos confinados, etiologia multifatorial, e pode causar prejuízo econômico. O objetivo do presente estudo foi determinar a capacidade da acidificação urinária mediante o uso de cloreto de amônio em ovinos. Foram utilizados 25 ovinos de três meses de idade, confinados e divididos aleatoriamente em três grupos: grupo CG (controle) não recebeu cloreto de amônio; grupo G200 (200mg/kg/PV) recebeu cloreto de amônio por 56 dias consecutivos; grupo G500 (500mg/kg/PV) recebeu cloreto de amônio por 56 dias consecutivos, administrados diariamente por via oral. Os momentos (M) de colheita de amostras e de avaliação clínica foram realizados com intervalo de sete dias, sendo M1 (imediatamente antes do cloreto de amônio), M2 (sete dias após) até M9, totalizando 70 dias de confinamento. Foram realizadas hemogasometria, concentração sérica de ureia e creatinina e avaliação ultrassonográfica do trato urinário. Na urinálise, houve uma maior incidência de cristais de fosfato amônio magnesiano no início do estudo, com migração para formação de cristais de urato, principalmente no G500, devido à acidificação urinária. Não houve alterações hemogasométricas, na bioquímica sérica, no líquido ruminal, ou alterações ultrassonográficas. A acidificação urinária foi obtida e mantida a partir do M7 durante a administração do cloreto de amônio no grupo G500, não ocorrendo nos outros grupos de estudo.(AU)

Porosity and pore design influence on fatigue behavior of 3D printed scaffolds for trabecular bone replacement.

Polymeric scaffolds provide several advantages when compared with other bone replacement and regenerating techniques. Namely, when compared with the current gold standard, bone autografts, there is no shortage of supply nor donor site morbidity. Contrarily to metallic implants, their mechanical properties are similar to those of cortical bone and they are biodegradable, therefore stress shielding is not expected to occur, and they will be gradually replaced by new bone tissue. Yet, there are still several challenges to overcome. After implantation scaffolds are subjected to dynamic loads, thus understanding polymeric scaffolds' fatigue behavior plays a major role on the design of better products. In this work PLA scaffolds were manufactured using 3D printing with optimized parameters. A total of six configurations were tested under static and dynamic load conditions. Static compression testing and numerical simulation showed good correlation. Numerical simulation provided a viable resource for scaffold design and innovation. Four different low-cycle fatigue loads were applied, during 3600 cycles with a frequency of 0.25 Hz. While under dynamic conditions, with a maximum stress of 24 MPa and R = 0.1, the apparent compressive modulus reached 973 MPa, due to pore collapse. Even after 3600 cycles no significant fatigue damage mechanisms were found on low porosity scaffolds, rendering them useful for trabecular bone replacement under dynamic conditions.

Morphological and mechanical characterization of 3D printed PLA scaffolds with controlled porosity for trabecular bone tissue replacement.

Bone transplant is still the gold standard approach when dealing with orthopedic trauma or disease. When this solution is not possible, scaffolding is a possibility provided by bone tissue engineering. To support the regeneration process, damaged bone tissue is removed and replaced by porous scaffold structures. In recent years, additive manufacturing has shown huge potential to produce scaffold structures with the required performance. In the current work, PLA scaffolds with different designs were 3D printed, using optimal manufacturing parameters. Scaffolds with three different porosity values were obtained by changing the filament offset from 571 to 1333 µm. A total of twelve designs were tested under monotonic and dynamic compression conditions. Numerical analysis showed good correlation with experimental results, allowing for a better assessment of scaffold mechanical behavior. Stress relaxation was measured on four different strain levels, assessing scaffold's behavior after implantation and consequent static response over time. Overall, orthogonal design provided better performance, due to improved material deposition. With lower porosity scaffolds equilibrium stress reached 24 MPa after 300 s relaxation time under 4% deformation, and the obtained equilibrium modulus was 428 MPa. Overall, attained results show that 3D printing with PLA can be applied in the manufacture of scaffolds for trabecular bone replacement.

Avaliação da maturidade pulmonar de cabritos nascidos a termo e prematuros / [Evaluation of lung maturity of term and premature kid]

Avaliou-se a maturidade pulmonar de cabritos no líquido amniótico de suas mães pela coloração de Shor, pelo azul de Nilo e pela contagem de corpos lamelares, bem como a vitalidade e os níveis de glicose e lactato séricos em cabritos nascidos a termo e prematuros. Para tanto, foram utilizados 32 cabritos, divididos em três grupos, a saber: grupo I: cabritos nascidos de cesarianas com 149 dias de gestação; grupo II: cabritos nascidos de cesarianas com 143 dias de gestação; e grupo III: cabritos nascidos de cesarianas com 143 dias de gestação, oriundos de mães que receberam, por via intramuscular, 20mg/cabra de dexametasona, 36 horas antes da cirurgia eletiva. A coloração de Shorr e a contagem de corpos lamelares demonstraram ser métodos diagnósticos promissores para a avaliação da maturidade pulmonar em neonatos caprinos. Contudo, a administração de dexametasona nas cabras no período antenatal não influenciou a maturidade fetal. Constatou-se, entretanto, que a avaliação física do paciente, logo após o nascimento, também se mostra fundamental no que tange à percepção da vitalidade e da viabilidade de cabritos recém-nascidos.(AU)

Pulmonary maturity of goats in their amniotic fluid was evaluated by Shor, Nile blue staining, and lamellar body count, as well as vitality and serum glucose and lactate levels in term and premature goats. Twenty-four kids were divided into three groups: group I: comprised of eight animals born by cesarean section with 149 days of gestation; group II: comprised of eight animals born by cesarean section with 143 days of gestation; and group III: comprised of eight animals born by cesarean section with 143 days of gestation, in which the does received intramuscular dexamethasone (20mg/goat) 36 hours prior to elective cesarean section. Shorr staining and lamellar body count have shown to be promising diagnostic methods for the assessment of lung maturity in goat neonates. However, the administration of dexamethasone to goats during antenatal period did not influence fetal maturity. It was verified that the physical evaluation of the patient, shortly after birth, is fundamental for the perception of vitality and viability of newborn goats.(AU)

Ocorrência de espécies do gênero Eimeria Schneider, 1875, em cordeiros confinados / Occurrence of Eimeria Schneider, 1875 species in feedlot lambs

Este trabalho descreve as espécies do gênero Eimeria Schneider, 1875, que ocorreram em um confinamento de cordeiros, bem como as dinâmicas da eliminação de oocistos no ambiente, a correlação com o ganho de peso médio diário (GMD) e as variáveis climáticas, durante nove semanas. Cento e quatro cordeiros de diversas raças e cruzas, com aproximadamente 60 dias de vida, foram confinados e submetidos a pesagens e avaliações clínicas e coprológicas periódicas. Amostras de fezes com mais de 500 oocistos de Eimeria por grama de fezes (OoPG) foram separadas para esporulação e identificação das espécies. Entre os oocistos avaliados, foram identificadas as espécies: E. parva, E. crandallis, E. ovinoidalis, E. weybridgensis, E. bakuensis, E. marsica, E. ahsata, E. granulosa, E. pallida e E. faurei. Eimeria crandallis foi a mais frequente, presente em 44 das 58 amostras avaliadas, enquanto E. parva foi a mais abundante nas contagens individuais. Nenhum dos animais apresentou quadro de eimeriose, e coeficientes negativos foram encontrados nas correlações OoPG vs. GMD (-0,075) e OoPG vs. pluviosidade (-0,1164), enquanto para OoPG vs. temperatura foi encontrado coeficiente positivo (0,2914). Animais positivos para a eliminação de oocistos apresentaram infecção mista nas avaliações semanais, com até sete espécies parasitando um mesmo cordeiro.(AU)

This study describes the Eimeria Schneider, 1875 species that occurred in a lamb feedlot, as well as the dynamics of oocyst output in the environment and its correlation with daily weight gain (DWG) and climatic variables during nine weeks. One hundred and four lambs of various breeds and crossbreeds, at approximately 60 days old, were placed in a feedlot and submitted to periodic weighing and clinical and coprological evaluations. Fecal samples presenting more than 500 Eimeria spp. oocysts per gram (OPG) were separated for sporulation, and oocysts were identified at species level. Among evaluated oocysts, the following species were identified: E. parva, E. crandallis, E. ovinoidalis, E. weybridgensis, E. bakuensis, E. marsica, E. ahsata, E. granulosa, E. pallida and E. faurei. Eimeria crandallis was the most frequent one, being identified in 44 of the 58 evaluated samples, while E. parva was the more abundant in individual counts in most weeks. None of the animals presented clinical signs of eimeriosis and negative correlation coefficients were found for OPG vs DWG (-0.075) and OPG vs rainfall, as for OPG vs temperature this coefficient was positive. Animals shedding oocysts presented mixed infection, with up to seven species parasitizing the same lamb.(AU)

Circulating ß cell-specific CD8+ T cells restricted by high-risk HLA class I molecules show antigen experience in children with and at risk of type 1 diabetes.

In type 1 diabetes (T1D), autoreactive cytotoxic CD8+ T cells are implicated in the destruction of insulin-producing ß cells. The HLA-B*3906 and HLA-A*2402 class I genes confer increased risk and promote early disease onset, suggesting that CD8+ T cells that recognize peptides presented by these class I molecules on pancreatic ß cells play a pivotal role in the autoimmune response. We examined the frequency and phenotype of circulating preproinsulin (PPI)-specific and insulin B (InsB)-specific CD8+ T cells in HLA-B*3906+ children newly diagnosed with T1D and in high-risk HLA-A*2402+ children before the appearance of disease-specific autoantibodies and before diagnosis of T1D. Antigen-specific CD8+ T cells were detected using human leucocyte antigen (HLA) class I tetramers and flow cytometry was used to assess memory status. In HLA-B*3906+ children with T1D, we observed an increase in PPI5-12 -specific transitional memory CD8+ T cells compared to non-diabetic, age- and HLA-matched subjects. Furthermore, PPI5-12 -specific CD8+ T cells in HLA-B*3906+ children with T1D showed a significantly more antigen-experienced phenotype compared to polyclonal CD8+ T cells. In longitudinal samples from high-risk HLA-A*2402+ children, the percentage of terminal effector cells within the InsB15-24 -specific CD8+ T cells was increased before diagnosis relative to samples taken before the appearance of autoantibodies. This is the first study, to our knowledge, to report HLA-B*3906-restricted autoreactive CD8+ T cells in T1D. Collectively, our results provide evidence that ß cell-reactive CD8+ T cells restricted by disease-associated HLA class I molecules display an antigen-experienced phenotype and acquire enhanced effector function during the period leading to clinical diagnosis, implicating these cells in driving disease.

Transferrina como marcador de maturação pulmonar em cordeiros nascidos a termo ou prematuros / Transferrin as a marker of pulmonary maturation in lambs born at term or premature

O objetivo do estudo foi procurar proteínas de fase aguda que possam indicar sinais de maturação no neonato prematuro, por meio da quantificação sérica delas. Identificou-se a imunoglobulina A, a ceruloplasmina, a haptoglobina, a glicoproteína ácida, a transferrina, a albumina e as imunoglobulinas G de cadeias leve e pesada, pela comparação do perfil dos proteinogramas de cordeiros nascidos a termo com os prematuros submetidos a diferentes protocolos terapêuticos, a fim de estimular a atividade respiratória. Constituíram-se seis grupos: PN (n= 9): nascidos de parto normal; CN (n= 7): nascidos de cesariana em tempo normal de gestação; CP (n= 6): nascidos de cesariana prematura sem nenhum tipo de tratamento; DEX (n= 9): prematuros cujas mães receberam dexametasona pré-parto; SURF (n= 6): prematuros tratados com surfactante; e DEXSURF (n= 6): prematuros tratados com surfactante cujas mães receberam dexametasona pré-parto. As avaliações foram realizadas nos momentos imediatamente após o nascimento (M0), após 24 (M24) e após 48 horas (M48). As amostras foram processadas por meio de eletroforese em gel de poliacrilamida contendo dodecil sulfato de sódio (SDS-PAGE). A albumina, as imunoglobulinas e a proteína total dos cordeiros tiveram elevação após a ingestão de colostro. Maiores valores séricos de transferrina são referentes a maior período gestacional, podendo essa proteína ser utilizada como marcador de maturação neonatal.(AU)

The aim of this study was to search for acute phase proteins that could indicate signs of maturation in the premature neonate by quantifying them in serum. Immunoglobulin A, ceruloplasmin, haptoglobin, acid glycoprotein, tranferrin, albumin, light and heavy chain immunoglobulin G were quantified, comparing the profile of proteinograms from term to preterm lambs submitted to different protocols that stimulate respiratory activity. Six groups were used: PN (n= 9): born from normal birth; CN (n= 7): born from caesarean section at normal time of gestation; CP (n= 6): born from premature cesarean without any type of treatment; DEX (n= 9) preterm whose mothers received prepartum dexamethasone; SURF (n= 6) preterm treated with surfactant; DEXSURF (n= 6): preterm treated with surfactant whose mothers received prepartum dexamethasone. The evaluations were performed immediately after birth (M 0), after 24 and 48 hours (M 24 and M 48). Samples were processed with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Albumin, immunoglobulins, and serum total protein of the lambs were elevated, after colostrum ingestion. Higher serum transferrin values refer to a longer gestational period, and this protein may be used as a marker of neonatal maturation.(AU)

A new hybrid approach for MHC genotyping: high-throughput NGS and long read MinION nanopore sequencing, with application to the non-model vertebrate Alpine chamois (Rupicapra rupicapra).

The major histocompatibility complex (MHC) acts as an interface between the immune system and infectious diseases. Accurate characterization and genotyping of the extremely variable MHC loci are challenging especially without a reference sequence. We designed a combination of long-range PCR, Illumina short-reads, and Oxford Nanopore MinION long-reads approaches to capture the genetic variation of the MHC II DRB locus in an Italian population of the Alpine chamois (Rupicapra rupicapra). We utilized long-range PCR to generate a 9 Kb fragment of the DRB locus. Amplicons from six different individuals were fragmented, tagged, and simultaneously sequenced with Illumina MiSeq. One of these amplicons was sequenced with the MinION device, which produced long reads covering the entire amplified fragment. A pipeline that combines short and long reads resolved several short tandem repeats and homopolymers and produced a de novo reference, which was then used to map and genotype the short reads from all individuals. The assembled DRB locus showed a high level of polymorphism and the presence of a recombination breakpoint. Our results suggest that an amplicon-based NGS approach coupled with single-molecule MinION nanopore sequencing can efficiently achieve both the assembly and the genotyping of complex genomic regions in multiple individuals in the absence of a reference sequence.

Real-world, long-term survival of incident patients with pulmonary arterial hypertension.

BACKGROUND: Pulmonary arterial hypertension (PAH) is a progressive, fatal disease. Long-term outcomes data are scarce in Portugal. We aimed to estimate survival of newly diagnosed PAH at a Portuguese referral center in the modern management era. METHODS: Between January 2009 and November 2015 all incident PAH cases were consecutively enrolled in a prospective cohort study. Sixty-five patients were followed up for a median of 3.1 [interquartile range 1.7-5.4] years. Kaplan-Meier survival analysis was used to estimate 1-, 3-, and 5-year survival and to compare it with a historical PAH survival estimated from the NIH cohort. RESULTS: Mean age was 48±19 years with female preponderance (68%). The most common PAH subgroup was congenital heart disease (PAH-CHD) (n=31; 48%), followed by connective tissue disease (PAH-CTD) (n=16; 25%), idiopathic (IPAH) (n=8; 12%) and hereditary (HPAP) (n=1; 1.5%). BNP values (hazard ratio [HR] 2.07; 95%CI 1.34-3.22; P=0.001) and male gender [HR 4.34 (1.44-13.09); P=0.009] were predictors of death. Survival rates at 1-, 3- and 5-years were 95%, 77% and 71%. Survival was not statistically different between PAH etiologies (Log-rank P=0.7). However, PAH-CHD was associated with a decreased risk of the combined endpoint of all-cause mortality and admission for decompensated heart failure [HR 0.36 (0.15-0.85); P=0.02]. We found a non-significant numerically higher survival of incident IPAH, HPAH and DPAH patients in comparison with the historical NIH cohort. CONCLUSIONS: In this cohort of incident PAH patients, PAH-CHD patients had better overall prognosis. Higher BNP values and male gender were associated with higher mortality.

Growth and hematological responses of tambaqui fed different amounts of cassava (Manihot esculenta) / Crescimento e respostas hematológicas de tambaquis alimentados com diferentes quantidades de mandioca (Manihot esculenta)

Tambaqui,Colossoma macropomum is a fish of primary importance in Brazilian aquaculture and in the Amazon region in particular. The aim of this work is to analyze the combined effects of physical training and levels of dietary cassava (Manihot esculenta) on the hematological parameters, food intake, conversion efficiency, growth ratio and swimming performance of this fish. A diet for tambaqui consisting of 30% cassava caused decreases in weight gain and specific growth rate compared with the control group. Diets containing 15% or 45% cassava did not affect these indices and did not cause hematological changes in tambaqui juveniles, indicating that some amount of cassava can be used as an alternative energy source for this important aquaculture species.(AU)

Tambaqui Colossoma macropomum, é um peixe de importância fundamental na aquicultura do Brasil e, em particular, na Região Amazônica. O objetivo deste trabalho foi analisar o efeito combinado do treinamento físico com diferentes níveis de macaxeira (Manihot esculenta) na dieta, sobre os parâmetros hematológicos, ingestão alimentar, eficiência de conversão, taxa de crescimento e performance natatória desses peixes. A dieta com 30% de cassava causou diminuição no ganho de peso e na taxa de crescimento específico quando se comparou ao grupo controle. Dietas contendo 15% e 45% de cassava não apresentaram efeito sobre estes índices e não causaram alterações hematológicas significativas em indivíduos juvenis de tambaqui, indicando que estas quantidades de cassava podem ser administradas como alternativa de fonte de energia para essa importante espécie na aquicultura.(AU)

Acute hypoxia up-regulates HIF-1α and VEGF mRNA levels in Amazon hypoxia-tolerant Oscar (Astronotus ocellatus).

Amazon fish maintain oxygen uptake through a variety of strategies considered evolutionary and adaptive responses to the low water oxygen saturation, commonly found in Amazon waters. Oscar (Astronotus ocellatus) is among the most hypoxia-tolerant fish in Amazon, considering its intriguing anaerobic capacity and ability to depress oxidative metabolism. Previous studies in hypoxia-tolerant and non-tolerant fish have shown that hypoxia-inducible factor-1α (HIF-1α) gene expression is positively regulated during low oxygen exposure, affecting vascular endothelial growth factor (VEGF) transcription and fish development or tolerance in different manners. However, whether similar isoforms exists in tolerant Amazon fish and whether they are affected similarly to others physiological responses to improve hypoxia tolerance remain unknown. Here we evaluate the hepatic HIF-1α and VEGF mRNA levels after 3 h of acute hypoxia exposure (0.5 mgO2/l) and 3 h of post-hypoxia recovery. Additionally, hematological parameters and oxidative enzyme activities of citrate synthase (CS) and malate dehydrogenase (MDH) were analyzed in muscle and liver tissues. Overall, three sets of responses were detected: (1) as expected, hematocrit, hemoglobin concentration, red blood cells, and blood glucose increased, improving oxygen carrying capacity and glycolysis potential; (2) oxidative enzymes from liver decreased, corroborating the tendency to a widespread metabolic suppression; and (3) HIF-1α and VEGF increased mRNA levels in liver, revealing their role in the oxygen homeostasis through, respectively, activation of target genes and vascularization. This is the first study to investigate a hypoxia-related transcription factor in a representative Amazon hypoxia-tolerant fish and suggests that HIF-1α and VEGF mRNA regulation have an important role in enhancing hypoxia tolerance in extreme tolerant species.

Multi-channel audio-based estimation of the Pre-Ejection Period.

Systolic time intervals (STI) have significant diagnostic and prognostic value to assess the global cardiac function. Presently, STIs are regarded as a promising tool for long-term follow-up of patients with chronic cardiovascular diseases. Heart sound has proven to be a valuable approach for STI estimation, in particular for the Pre-Ejection Period (PEP). However, since the optimal auscultation site varies from individual to individual, as well as with the position of the body, its application in single-channel and fixed auscultation site setups poses practical difficulties. Hence, we extend our previous work on PEP estimation to a multi-channel sound acquisition setup, where signal redundancy is exploited. A channel selection method is proposed and the best channel is selected for PEP estimation. As a preliminary study, the devised algorithms were evaluated with respect to echocardiography reference on a set of 236 heartbeats collected from 8 healthy subjects in two sound auscultation sites. The channel selection approach led to 8.4% estimation error decrease, in comparison to a single-channel approach. Current results support our assumption that a multi-channel audio-based strategy can be applied to assess STI in personal health application scenarios.

Reproducibility of peak power output during a 10-s cycling maximal effort using different sampling rates.

The study was aimed to investigate the reproducibility of performance parameters obtained from 10-s maximal cycling effort against different braking forces in young adult athletes. The sample (n = 48) included male athletes aged 18.9-29.9 years (175.5 ± 6.9 cm, 76.2 ± 10.1 kg). The exercise protocol was performed in a cycle-ergometer against a random braking force (4% to 11% of body mass). Intra-individual variation was examined from repeated tests within one week. Descriptive statistics were computed and differences between sessions were tested using paired t-test. The coefficient of correlation between repeated measures, technical error of measurement (TEM), coefficient of variation and ICC were calculated. Agreement between trials was examined using the Bland-Altman procedure. Mean values of peak power were relatively stable when obtained from sampling rates of 50 Hz and ranged between 1068 watt and 1082 watt (t(47) = 1.149, p = 0.256, ES-r = 0.165) or while corresponding to a sampling rate of 1 Hz (t(47) = 0.742, p = 0.462, ES-r = 0.107). Correlations between repeated measures were high (+0.907, 95% CI: +0.839 to +0.947) and TEM about 59.3 watt (%CV = 5.52%; ICC = 0.951, 95% CI: 0.912 to 0.972). The present study suggests that reproducibility of peak power in male adult athletes tended to be acceptable and within individual error appeared unrelated to braking force.

Effect of high-dose irradiation on quality characteristics of ready-to-eat broiler breast fillets stored at room temperature.

The effect of high-dose irradiation on the physical, chemical, and bacteriological parameters of ready-to-eat vacuum-packed broiler breast meat after 430 d of storage at room temperature was investigated. Ready-to-eat broiler breast fillets were immersed in brine with garlic powder and then drained, grilled, and vacuum-packed (primary packaging). The high-dose irradiation used was approximately 48 kGy. The treatments were designated as A (irradiated samples stored at room temperature), B (irradiated samples stored at -25°C), and C (nonirradiated samples stored at -25°C). All samples were packaged in polyethylene bags containing aluminum to exclude light (secondary packaging). Proximate composition, pH, 2-thiobarbituric acid reactive substance (TBARS), and heterotrophic aerobic mesophilic bacteria were analyzed during 430 d of storage. Results were analyzed using 1-way ANOVA and the Tukey test. Linear regression was used to analyze the correlation between the results for each parameter and storage time of the different treatments. The gamma radiation caused slight changes (P < 0.05) in the moisture and fat content, regardless of storage temperature. After storage d 110, TBARS values remained stable (P > 0.05) in all the treatments. The preservation methods used were effective in maintaining the mesophilic counts below the detection level during the entire storage period. We concluded that, among the treatments studied, high-dose irradiation with storage at room temperature showed potential for the preservation of ready-to-eat products made from poultry meat, to provide foods safe for consumption.

Microbiological quality and biogenic amines in ready-to-eat grilled chicken fillets under vacuum packing, freezing, and high-dose irradiation.

The combined effects of cooking, vacuum packing, freezing, and high-dose gamma irradiation in the microbiological conservation and in biogenic amine (BA) contents of ready-to-eat grilled breast chicken fillets are investigated in this work. After seasoning, cooking, and vacuum packing, one-third of the samples were stored at -25°C (T1). The remaining two-thirds were treated with 48 kGy, one-third being stored at -25°C (T2) and the other one-third kept at room temperature (T3). All samples were periodically analyzed to determine growth of heterotrophic aerobic mesophilic bacteria (HAMB) and levels of BA (tyramine, TYM; putrescine, PUT; cadaverine, CAD; spermidine, SPD; histamine, HYM; and spermine, SPM). Variance analysis was performed to determine significant changes in the measured data. Grilling caused HAMB counts in seasoned samples to drop from 5.3 log cfu/g to zero. In addition, no viable HAMB cells were detected in the samples throughout the 12-mo storage time. Regarding the BA analyses, the highest mean levels were measured for SPM and CAD with significantly higher levels (P < 0.05) being determined in nonirradiated samples (T1). Furthermore, significantly lower mean levels for the total content of BA were observed in the irradiated samples. Relative to T1 (7.5 ± 1.5 mg/kg), the figures were 47 ± 23% for T2 and 60 ± 25% for T3, mostly due to loss of CAD by radiolysis. Therefore, it can be concluded that the combination of grilling, vacuum packing, freezing, and high-dose gamma irradiation efficiently eliminated HAMB, while sustaining acceptable levels of BA in ready-to-eat chicken breast fillets throughout the 12 mo of storage at room temperature.

Case report: A novel TET2 mutation in a patient with refractory cytopenia with multilineage dysplasia.

Myelodysplastic syndrome diagnosis of karyotypically normal patients may be elusive because it relies exclusively on morphological and clinical data. In routine practice, finding of an acquired mutation or a cytogenetic abnormality provides irrefutable evidence of the clonal nature of that disease. Recurrent deletions and somatic mutations in TET2, a gene involved in epigenetic regulation, have been reported in about 20% of adult patients with myelodysplastic syndrome. We report a novel g.95805C>T, nonsense TET2 mutation, leading to a premature stop codon (p.Gln913*), in an adult patient with refractory cytopenia with multilineage dysplasia.

Association of social network and social support with health-related quality of life and fatigue in long-term survivors of Hodgkin lymphoma.

PURPOSE: As the number of survivors of Hodgkin's lymphoma (HL) increases, there has been a growing interest in long-term treatment-related side effects and their impact on the quality of life (QoL). The aim of this study was to assess the association of social network and social support with the QoL and fatigue among long-term HL survivors. METHODS: A total of 200 HL survivors were included. The generic Short Form-12 (SF-12) questionnaire, the QoL cancer survivor's questionnaire (QOL-CS), and the Multidimensional Fatigue Inventory were used to assess QoL and fatigue. Social network and social support were evaluated with the Social Support Survey. RESULTS: Social network and all social support measures were favorably associated with two or more SF-12 scales, mainly with physical functioning and the mental health scales. Social network and social support dimensions were also associated with better QOL-CS scores. Affective support, informational support, positive interaction, and emotional support were associated with less fatigue. CONCLUSIONS: Both social network and social support are associated with better QoL and lower levels of fatigue in HL survivors. This information may be useful to health professionals and community organizations in implementing effective interventions to improve these patients' quality of life.

Application of the intelligent techniques in transplantation databases: a review of articles published in 2009 and 2010.

The replacement of defective organs with healthy ones is an old problem, but only a few years ago was this issue put into practice. Improvements in the whole transplantation process have been increasingly important in clinical practice. In this context are clinical decision support systems (CDSSs), which have reflected a significant amount of work to use mathematical and intelligent techniques. The aim of this article was to present consideration of intelligent techniques used in recent years (2009 and 2010) to analyze organ transplant databases. To this end, we performed a search of the PubMed and Institute for Scientific Information (ISI) Web of Knowledge databases to find articles published in 2009 and 2010 about intelligent techniques applied to transplantation databases. Among 69 retrieved articles, we chose according to inclusion and exclusion criteria. The main techniques were: Artificial Neural Networks (ANN), Logistic Regression (LR), Decision Trees (DT), Markov Models (MM), and Bayesian Networks (BN). Most articles used ANN. Some publications described comparisons between techniques or the use of various techniques together. The use of intelligent techniques to extract knowledge from databases of healthcare is increasingly common. Although authors preferred to use ANN, statistical techniques were equally effective for this enterprise.