Sacroiliac joint dysfunction and lumbar pain. Evaluation in a Brazilian population.

The study intended to evaluate the incidence and evolution of sacroiliac joint dysfunction (SIJD). To reach 50 patients with SIJD diagnosis, 192 patients with low back pain and failure in conservative approach were consecutively examined (26% incidence). Initially patients underwent intra-articular (IA) corticosteroid sacroiliac joint (SIJ) block followed, if necessary, by cooled SIJ radiofrequency or referred to surgical intervention, in order of complexity. From the 50 patients submitted to IA SI block, 41 (82%) referred pain and quality of life improvement and lesser rescue analgesics consumption for 25 weeks. The block induced a prompt onset of pain relief and there was a drop in mean pain score from 8 to 2 cm (p < 0.001) maintained up to 25 weeks. Rescue analgesic consumption also significantly dropped (p < 0.05). However, nine patients (18%) did not refer long lasting improvement in the third week evaluation and underwent cooled radiofrequency. From this population of nine, seven were successful (78%) while two were recommended surgery. In view of the 50 patients, 82% were comfortable after IA block, 18% were submitted to radiofrequency, with a success rate of 78%. The final incidence of surgery suggestion was 4%.

El estudio pretende evaluar la incidencia y evolución de la disfunción de la articulación sacroilíaca (DASI). Para llegar a 50 pacientes con diagnóstico de DASI, se examinaron consecutivamente 192 pacientes con dolor lumbar y fracaso en el abordaje conservador (26% de incidencia). Inicialmente, los pacientes se sometieron a un bloqueo de la articulación sacroilíaca (ASI) con corticosteroides intraarticulares (IA) seguido, si era necesario, de radiofrecuencia ASI enfriada o remitidos a una intervención quirúrgica, en orden de complejidad. De los 50 pacientes sometidos al bloqueo IA SI, 41 (82%) refirieron mejoría del dolor y de la calidad de vida y menor consumo de analgésicos de rescate durante 25 semanas. El bloqueo indujo un rápido inicio del alivio del dolor y hubo una caída en la puntuación media del dolor de 8 a 2 cm (p < 0.001) mantenida hasta 25 semanas. El consumo de analgésicos de rescate también disminuyó significativamente (p < 0.05). Sin embargo, nueve pacientes (18%) no refirieron una mejoría duradera en la evaluación de la tercera semana y se sometieron a radiofrecuencia fría. De esta población de nueve, siete tuvieron éxito (78%), mientras que a dos se les recomendó cirugía. De los 50 pacientes, 82% se sintió cómodo después del bloqueo IA, 18% fue sometido a radiofrecuencia, con una tasa de éxito de 78%. La incidencia final de sugerencia de cirugía fue de 4%.

Methylene blue-mediated Photodynamic Therapy in human retinoblastoma cell lines.

Retinoblastoma is a malignant tumor of the retinal precursor cells and one of the rarest types of pediatric tumor, often occurring in the earliest years of life. Symptoms are conditioned by tumor size and location; one of the most recurrent symptoms is a white reflex in the pupillary area, called leukocoria or cat's eye reflex. In the present work, we studied the in vitro effectiveness of Photodynamic treatment (Pdt) in two types of human retinoblastoma, Y79 and WERI-Rb cell lines, using methylene blue (MB), a photosensitizer (PS) from the phenothiazine group. The two cell lines were incubated with varying concentrations of MB (3, 7, 10, 15, 20, 25, 30, 40, and 50 µM), in the absence of light (dark cytotoxicity) and, in the presence of 664 nm laser light (phototoxicity) with fluences of 1, 1.5, 3, 5, 7, 10, and 15 J/cm2. The Y79 cell line showed higher cellular uptake values for MB than the WERI-Rb cell line. After three hours of incubation, the Y79 and WERI-Rb took up 48% and 34% of the total photosensitizer present in the medium, respectively. Using MTT assay, the results showed that the Y79 cell line was more affected by the photo treatment as demonstrated by the combination of MB concentration and light doses compared with WERI-Rb cell line. The results were correlated with the more pronounced singlet oxygen emission observed in Y79 cells. While MB does show efficacy for eradication of retinoblastoma in vitro, only studies in appropriate animal models will reveal whether the selectivity of photokilling at tolerable drug and light doses is sufficient to suggest clinical trials.

Yellow grease in sheep diets: intake and digestibility / [Óleo residual de fritura em dietas para ovinos: consumo e digestibilidade]

This study aimed to assess the effects of yellow grease supplementation on the intake, digestibility, and nitrogen balance in sheep. Twenty Santa Inês lambs with a mean age of 95 ± 10 d and body weight of 19.29 ± 3.17kg were evaluated in a completely randomized design. The diets were supplemented with oil at concentrations of 0, 20, 40, 60, and 80 gkg-1 of dry matter (DM) of the concentrate. The diets were based on roughage and concentrate (50:50). The experimental period lasted 19 d and included 14 adaptation days and five collection days for the total supplied diet, orts, feces, and urine. Supplementation with yellow grease had no significant effect on the intake of DM, crude protein (CP), neutral detergent fiber (NDF), or non-fiber carbohydrates (NFC). However, the ether extract (EE) intake increased linearly with supplementation of yellow grease. Moreover, no effect was observed for DM, CP, NDF, and NFC digestibility and nitrogen balance. EE digestibility increased linearly with the yellow grease dietary supplementation. Thus, sheep dietary supplementation with yellow grease may be used at a level of up to 80 gkg-1 of DM of concentrate without impairing nutrient intake and digestibility.(AU)

Objetivou-se, com o estudo, avaliar os efeitos do óleo residual de fritura, em dietas para ovinos, sob o consumo, a digestibilidade e o balanço de nitrogênio. Foram utilizados 20 cordeiros Santa Inês, com idade de 95 ± 10 dias e peso corporal de 19,29 ± 3,17kg, em delineamento inteiramente ao acaso. As dietas continham óleo de fritura nas concentrações de 0; 20; 40; 60 e 80gkg-1 da matéria seca (MS) do concentrado. As dietas tinham relação volumoso:concentrado de 50:50. O período experimental foi de 19 dias, incluindo 14 dias em adaptação e cinco dias de coleta do fornecido, das sobras, das fezes e da urina. A suplementação com óleo de fritura não alterou o consumo de MS, proteína bruta (PB), matéria orgânica (MO), fibra em detergente neutro (FDN) e carboidratos não fibrosos (CNF). Entretanto, o consumo de extrato etéreo (EE) aumentou com a inclusão do óleo. Não foi observado efeito na digestibilidade da MS, da PB, da FDN, dos CNF e no balanço de nitrogênio. A digestibilidade do EE aumentou com a inclusão do óleo. Assim, a inclusão de óleo de fritura em dietas para ovinos pode ser utilizada em até 80gkg-1 da MS do concentrado, sem limitar ingestão e digestibilidade dos nutrientes.(AU)

Quantificação do DNA e genotipagem de biópsias de embriões bovinos Gir e Girolando produzidos in vitro / DNA quantification and genotyping of biopsies obtained from Gyr and Girolando in vitro produced bovine embryos

A biópsia embrionária associada à genotipagem permite a obtenção de informações genômicas antes mesmo da transferência dos embriões. Neste estudo, foram avaliadas amostras biopsiadas de blastocistos bovinos transferidos para receptoras (n=47), sob a hipótese de que a raça (Gir ou Girolando), o estádio embrionário (blastocisto ou blastocisto expandido) e a competência para estabelecimento de prenhez (positiva ou negativa) afetariam a quantidade e a qualidade do DNA da amostra obtida. O DNA foi extraído, amplificado, quantificado por eletroferograma e genotipado. O parâmetro call rate (CR) foi adotado para mensurar a qualidade da genotipagem. Obteve-se concentração de DNA de 86,07±171,66ng/µL e CR 0,73±0,17. O CR não variou em função da quantidade de DNA nas amostras. As variáveis raça e estádio embrionário não influenciaram a concentração de DNA, nem o CR. Houve efeito da prenhez sobre o CR (P=0,0187), mas, como houve maior CR nas amostras provenientes do grupo prenhez negativa, não foi possível associar esse parâmetro à qualidade embrionária. Concluiu-se que a raça e a qualidade embrionária não afetam os parâmetros aqui estudados em amostras embrionárias, ou seja, embriões com maiores chances de implantação não refletem alta qualidade nas amostras de biópsia genotipadas.(AU)

Embryo biopsy associated with genotyping allows genomic information before embryo transfer. In this study, blastocyst biopsy samples from embryos transferred to recipients (n= 47) were evaluated, under the hypothesis that breed (Gyr or Girolando), embryonic stage (blastocyst or expanded blastocyst) and competence to establish pregnancy (positive or negative) would affect the quantity and DNA quality of samples. DNA was extracted, amplified, quantified by electropherogram and genotyped. The parameter call rate (CR) was used to measure the quality of genotyping. DNA concentration of 86.07±171.66ng/µl, and CR 0.73±0.17 was obtained. CR did not vary according to the amount of DNA in the samples. The variables breed and embryonic stage had no influence on DNA concentration or CR. There was pregnancy effect on the CR (P= 0.0187), but since there was a higher CR in the samples from the negative pregnancy group, it was not possible to associate this parameter with the embryonic quality. We conclude that the breed and embryo quality do not affect the evaluated parameters in embryonic samples. Embryos with higher chances of implantation do not reflect high quality in embryo biopsy genotyped samples.(AU)

Influência da dificuldade de inseminação, temperamento e cortisol plasmático sobre a taxa de concepção de vacas e novilhas da raça Nelore inseminadas em tempo fixo / Influence of difficulty of insemination, temperament and plasma cortisol on conception rate of Nelore cows and heifers in timed AI

O objetivo do presente experimento foi avaliar efeito do estresse e da dificuldade de inseminação (DifIA) sobre a taxa de concepção (TC) de vacas (n=93) e novilhas (n= 72) Nelore submetidas à IATF. No D9, anotou-se nota de temperamento (NTe) e tempo da saída do brete (TSB) de todos os animais e coletou-se sangue das novilhas. No dia da IATF (D11), anotou-se NTe, TSB, DifIA e tempo de IA. A TC foi 36% para vacas e 46% para novilhas (P>0,05). Não foi observado efeito de Nte sobre TC (P>0,05). Porém, houve tendência para maior (P<0,10) TC nos animais que não apresentaram dificuldade de inseminação (DifIA1; TP=42%) em comparação aos animais com moderada ou alta dificuldade (DifIA2+DifIA3; TP=27%). Foi observado menor (P<0,05) tempo de IA para animais DifIA1 (17:31±06:02s) que animais DifIA2-3 (30:10±15:45s). Novilhas com maiores (P<0,05) níveis de cortisol apresentaram maior NTe (P<0,05). Entretanto, TC (59%) das novilhas menos reativas (cortisol=4,12±1,12ng/mL; NTe=3,2±0,6) não diferiu da TC (41%; P>0,05) das mais agitadas (cortisol=7,76±1,33ng/mL; NTe=3,82±0,79). Concluiu-se que avaliações de temperamento se relacionaram com nível de estresse, embora esses parâmetros não tenham afetado a TC deste trabalho. A maior dificuldade e/ou tempo necessário para se completar a IA demonstrou ser um potencial fator para a redução da fertilidade na IATF.(AU)

The objective was to evaluate the influence of stress and difficulty of insemination (DifIA) on conception rate (CR) of Nellore cows (n= 93) and heifers (n= 72) in Timed-AI. On D9, temperament (NTe) and time for chute exit (TSB) were recorded for all animals, and blood samples were colected from heifers. On the day of Timed-AI (D11), NTe, TSB, DifIA and time for AI were recorded. For cows, CR was 36% and for heifers 46% (P> 0.05). No effect (P> 0.05) of NTe was observed on CR. However, a tendency (P< 0,10) for higher CR was observed in animals with no difficulty for insemination (DifIA1; CR=42%) compared to animals that presented moderate or high difficulty (DifIA2+DifIA3, CR=27%). Time required for AI was lower (P< 0.05) in animals DifIA1 (17:31±06:02sec) than in animals DifIA2-3 (30:10±15:45sec). Heifers with greater (P< 0.05) cortisol levels presented higher Nte (P< 0.05). However, CR (59%) of less reactive heifers (cortisol=4,12±1,12ng/mL; NTe=3,2±0,6) did not differ from CR (41%; P> 0.05) of stressed animals (cortisol=7,76±1,33ng/mL; NTe=3,82±0,79). It was concluded that assessments of temperament were related to stress level, although these parameters did not affect the CR of this study. However, the higher difficulty and/or time to complete AI showed to be a potential factor for reducing fertility after timed-AI.(AU)

One for all and all for one: retention of colour-unchanged old flowers increases pollinator attraction in a hermaphroditic plant.

Long-lived flowers increase pollen transfer rates, but these entail high water and carbon maintenance costs. The retention of pollinated and reward-free old flowers enhances pollinator visitation to young receptive flowers by increasing floral display size. This mechanism is associated with acropetal inflorescences or changes in flower colour and openness, but the retention of unchanging solitary flowers remains overlooked. We examined pollination-dependent variation in floral longevity and determined stigmatic receptivity, pollen viability and pollen removal rates among flower ages in Kielmeyera regalis, a Neotropical savanna shrub. We also evaluated the effects of floral display size on pollinator visitation rates. Lastly, we determined whether old flowers are unvisited and exclusively increase pollinator attraction to young flowers through flower removal experiments. Regardless of pollination treatment, flowers lasted fully open with no detectable physical changes for 3 days. Over time, stigmas remained receptive but >95% of pollen was removed. Pollinator visitation significantly increased with floral display size and intermediate percentages (15-30%) of newly opened flowers. Accordingly, the retention of reward-free and unvisited old flowers increased young flower-pollinator interaction. Our results reveal the importance of a prolonged floral longevity in increasing pollinator attraction toward newly opened receptive flowers without changes in flower colour and form. We conclude that the retention of pollinated, reward-free and unvisited colour-unchanged old flowers in K. regalis is a strategy that counteracts the water use costs associated with the maintenance of large flowers with increased mate opportunities in a pollen-limited scenario.

Circulating levels of adiponectin and extent of coronary artery disease in patients undergoing elective coronary angiography

Adiponectin (APN), an adipose tissue-released adipokine with demonstrated anti-inflammatory and anti-atherogenic properties, is encoded by a gene whose polymorphisms are associated with presence of coronary artery disease (CAD). Serum APN levels are inversely related with presence and complexity of CAD. Within this context, we sought to compare levels of total APN and its high molecular weight form (HMW APN) according to clinical presentation and extent of CAD in patients undergoing elective cardiac catheterization. From March 2008 to June 2010, clinical data and blood samples for APN and HMW APN measurements were collected from 415 subjects undergoing cardiac catheterization at two tertiary centers. CAD extent was estimated by the number of coronary arteries with significant stenosis (≥70% obstruction in a major coronary artery) and by Duke Jeopardy Score (DJS). Serum APN levels were similar between groups with stable or unstable CAD (APN 9.20±5.88 vs 9.47±6.23 μg/mL, P=0.738, and HMW APN 5.31±3.72 vs 5.91±4.16 μg/mL, P=0.255), even after stratification by the number of arteries involved (single-vessel vs multivessel disease: APN 9.39±5.76 vs 9.26±6.27 μg/mL, P=0.871; HMW APN 5.29±3.79 vs 5.83±4.04 μg/mL, P=0.306) and DJS score (APN, P=0.718; HMW APN, P=0.276). We conclude that APN and HMW APN serum levels are similar across clinical presentations and different extents of CAD, despite being significantly lower in the presence of obstructive CAD.

Circulating levels of adiponectin and extent of coronary artery disease in patients undergoing elective coronary angiography.

Adiponectin (APN), an adipose tissue-released adipokine with demonstrated anti-inflammatory and anti-atherogenic properties, is encoded by a gene whose polymorphisms are associated with presence of coronary artery disease (CAD). Serum APN levels are inversely related with presence and complexity of CAD. Within this context, we sought to compare levels of total APN and its high molecular weight form (HMW APN) according to clinical presentation and extent of CAD in patients undergoing elective cardiac catheterization. From March 2008 to June 2010, clinical data and blood samples for APN and HMW APN measurements were collected from 415 subjects undergoing cardiac catheterization at two tertiary centers. CAD extent was estimated by the number of coronary arteries with significant stenosis (≥70% obstruction in a major coronary artery) and by Duke Jeopardy Score (DJS). Serum APN levels were similar between groups with stable or unstable CAD (APN 9.20±5.88 vs 9.47±6.23 µg/mL, P=0.738, and HMW APN 5.31±3.72 vs 5.91±4.16 µg/mL, P=0.255), even after stratification by the number of arteries involved (single-vessel vs multivessel disease: APN 9.39±5.76 vs 9.26±6.27 µg/mL, P=0.871; HMW APN 5.29±3.79 vs 5.83±4.04 µg/mL, P=0.306) and DJS score (APN, P=0.718; HMW APN, P=0.276). We conclude that APN and HMW APN serum levels are similar across clinical presentations and different extents of CAD, despite being significantly lower in the presence of obstructive CAD.

Chemical Speciation of Selenium and Mercury as Determinant of Their Neurotoxicity.

The antagonism of mercury toxicity by selenium has been well documented. Mercury is a toxic metal, widespread in the environment. The main target organs (kidneys, lungs, or brain) of mercury vary depending on its chemical forms (inorganic or organic). Selenium is a semimetal essential to mammalian life as part of the amino acid selenocysteine, which is required to the synthesis of the selenoproteins. This chapter has the aim of disclosing the role of selenide or hydrogen selenide (Se-2 or HSe-) as central metabolite of selenium and as an important antidote of the electrophilic mercury forms (particularly, Hg2+ and MeHg). Emphasis will be centered on the neurotoxicity of electrophile forms of mercury and selenium. The controversial participation of electrophile mercury and selenium forms in the development of some neurodegenerative disease will be briefly presented. The potential pharmacological use of organoseleno compounds (Ebselen and diphenyl diselenide) in the treatment of mercury poisoning will be considered. The central role of thiol (-SH) and selenol (-SeH) groups as the generic targets of electrophile mercury forms and the need of new in silico tools to guide the future biological researches will be commented.

Study on the association of UGT1A9 gene c.98T>C polymorphism and mycophenolic acid plasma levels in renal transplant patients.

Mycophenolate mofetil (MMF) is a prodrug active only after its hydrolysis to mycophenolic acid (MPA). The UGT1A9 enzyme is of special interest since it is the main enzyme involved in the glucuronidation of MPA. Single nucleotide polymorphisms (SNPs) in the UGT1A9 gene may be responsible for individual differences in the pharmacokinetics of MMF. Expression levels and the activity of UGT1A9 may depend on the presence of some SNPs located in the gene promoter region (-2152C>T and -275T>A), as well as changes in the coding region (c.98T>C). The objective of this study was to evaluate the effect of allelic variants of the UGT1A9 c.98T>C polymorphism (rs72551330; g. 87289T>C) on MMF metabolism in renal transplant patients. MPA and MPA 7-O glucuronide (MPAG) levels were determined on plasma samples of kidney transplant patients (N = 39) by high-performance liquid chromatography using ultraviolet detection. DNA was isolated from leukocytes and stored at -20°C. The presence of SNPs was investigated using polymerase chain reaction, followed by amplicon sequencing. The analysis of the UGT1A9 c.98T>C polymorphism revealed that all study patients presented the TT genotype. Diverse MPA and MPAG plasma concentrations were detected, including therapeutic, subtherapeutic, and toxic levels. A standardized molecular method permitted identification of UGT1A9 c.98T>C polymorphism genotypes in the examined renal transplant patients. All individuals of the study group presented the same genotype (c.98TT) for that polymorphism. Thereby, no association between the c.98T>C polymorphism and MPA and MPAG plasma levels could be evaluated, despite different levels of these compounds being observed.

Moderate aerobic exercise on the recovery phase of gentamicin-induced acute kidney injury in rats.

INTRODUCTION: Acute kidney injury is a serious public health problem, especially in intensive care units, where patients may require dialysis support, resulting in 50% mortality. AIM: To evaluate the effects of moderate aerobic exercise on the recovery phase of acute kidney injury induced by gentamicin in rats. MAIN METHODS: Male adult Wistar rats were allocated into 4 groups: W10+R30, G10+R30, W10+EX30 and G10+EX30; W10 received water (gentamicin vehicle) and G10 received gentamicin for 10days; R30 remained resting and EX30 made exercise for 30days after gentamicin suspension. Training was performed on treadmill. Blood, 24h urine and kidneys were collected for renal function and oxidative stress, antioxidant, TGF-ß and histological analysis. KEY FINDINGS: Gentamicin treatment caused decreased renal function significant oxidative stress, reduced urinary nitric oxide and increased TGF-ß. G10+R30 presented partial recovery of metabolic data, renal function and lipoperoxidation levels, although they were still altered compared to W10+R30. Besides, we observed the presence of lymphomononuclear infiltrate in the kidneys of G10+R30. G10+EX30 vs G10+R30 showed additional improvement of all the mentioned parameters, showing at histology, regeneration of the tubule epithelium. SIGNIFICANCE: Our data suggest that moderate exercises could help in the recovery of metabolic parameters, renal function and structure on gentamicin-induced AKI, perhaps due to restoration of redox balance. This could protect the kidneys from further insults like challenges with nephrotoxic drugs or the aging per se.

Kinetics data from bovine sex-specific embryo development from three different bulls.

Here we present kinetics data from bovine sex-specific embryo development. Embryos were originated using sex-sorted semen from three different Nelore bulls, and semen from the same batch was used for X-and Y-chromosome spermatozoa sorting. Data was obtained for six time points (24, 48, 96, 120, and 144 h.p.i.). Analyses for each bull׳s embryos (1, 2 and 3) is presented for female and male groups separately. Also, grouped data analysis, considering bull and sex interaction, is shown. For further interpretation and discussion, see "Cell death is involved in sexual dimorphism during preimplantation development" (Oliveira et al., 2015 [1]).

Evolution of Socioeconomic Conditions and Its Relation to Spatial-Temporal Changes of Giardiasis and Helminthiasis in Amazonian Children.

This study analyzed the evolution of socioeconomic, sanitary, and personal factors as well as spatiotemporal changes in the prevalence of helminthiasis and giardiasis in urban Amazonian children between 2003 and 2011. Child age, lack of sanitation, and lack of access to bottled water were identified as significant associated factors for helminthiasis and giardiasis. There was an overall improvement in socioeconomic and sanitary conditions in the city resulting in decreased helminth prevalences from 12.42 to 9.63% between 2003 and 2010, but the prevalence increased to 15.03% in 2011 due to migratory movement and unstable sanitary conditions. As for Giardiasis, socioeconomic and environmental changes were not enough to reduce prevalence (16% in 2003 and 23% in 2011). Spatial analysis identified a significant cluster for helminthiasis in an area of poor housing conditions. Control programs in the Amazon need to target high-risk areas focusing changes in sanitation, water usage, and health education.

Cell death is involved in sexual dimorphism during preimplantation development.

In bovine preimplantation development, female embryos progress at lower rates and originate smaller blastocysts than male counterparts. Although sex-specific gene expression patterns are reported, when and how sex dimorphism is established is not clear. Differences among female and male early development can be useful for human assisted reproductive medicine, when X-linked disorders risk is detected, and for genetic breeding programs, especially in dairy cattle, which requires female animals for milk production. The aim of this study was to characterize the development of female and male embryos, attempting to identify sex effects during preimplantation development and the role of cell death in this process. Using sex-sorted semen from three different bulls for fertilization, we compared kinetics of bovine sex-specific embryos in six time points, and cell death was assessed in viable embryos. For kinetics analysis, we detected an increased population of female embryos arrested at 48 and 120h.p.i., suggesting this time points as delicate stages of development for female embryos that should be considered for testing improvement strategies for assisted reproductive technologies. Assessing viable embryos quality, we found 144h.p.i. is the first time point when viable embryos are phenotypically distinct: cell number is decreased, and apoptosis and cell fragmentation are increased in female embryos at this stage. These new results lead us to propose that sex dimorphism in viable embryos is established during morula-blastocyst transition, and cell death is involved in this process.

Fenologia e prospecção fitoquímica do jaborandi (Pilocarpus microphyllus Stapf ex Holmes) / Phenology and phytochemical prospection of Jaborandi (Pilocarpus microphyllus Stapf ex Holmes)

RESUMO O estudo fenológico tem como finalidade determinar o ritmo sazonal dos eventos do ciclo de vida da planta, como floração e frutificação. Estes eventos são determinados por uma série de fatores, como: alternância de períodos chuvosos ou não chuvosos, intensidade da radiação solar, entre outros. A fitoquímica tem por objetivos conhecer os constituintes químicos de espécies vegetais ou avaliar sua presença. O presente trabalho teve como objetivo realizar a caracterização fenológica e a prospecção fitoquímica de folhas de jaborandi. A área de estudo para a avaliação do material vegetal foi o Banco Ativo de Germoplasma de Jaborandi da Embrapa Amazônia Oriental, situada no município de Belém-PA. Os acessos escolhidos foram: Merck, cultivado a pleno sol e à sombra; Japonês e Bonal 4, cultivados a pleno sol. Os registros foram realizados diariamente por um período de 28 meses correspondendo a agosto de 2010 a dezembro de 2012, de cinco plantas/acesso e organizados para demonstração mensal, através de fichas com a numeração respectiva das plantas, com registro de presença ou ausência das fenofases, floração e frutificação. A determinação do peso seco das amostras coletadas dos acessos foi realizada no Laboratório de Agroindústria da Embrapa Amazônia Oriental, onde, após a triagem e remoção das impurezas, as folhas foram cortadas, pesadas, colocadas em bandejas de inox e secas em estufa com circulação mecânica (FANEM 320-SE), à temperatura de 45º C por 120 h. Em seguida, as amostras foram pesadas, trituradas e acondicionadas em sacos plásticos devidamente identificados e guardados sob refrigeração à temperatura de 10º C até o uso. Os extratos das plantas foram preparados utilizando-se 100 g de folhas secas de cada acesso, triturados e submetidos à extração hidroalcoólica (etanol 80%) em banho-maria sob refluxo, por aproximadamente 4 horas. Os extratos foram armazenados protegidos da luz na geladeira até o momento das análises. Foi analisada a presença das seguintes classes de substâncias químicas: ácidos orgânicos, açúcares redutores, polissacarídeos, proteínas e aminoácidos, taninos, catequinas, flavonoides, glicosídeos cardíacos, lactonas sesquiterpênicas, azulenos, carotenoides, esteroides e triterpenoides, depsídeos e depsidonas, derivados da cumarina, saponina espumídica, alcaloides, purinas, antraquinonas. Os resultados obtidos demonstraram que a espécie Pilocarpus microphyllus apresentou floração durante o ano todo e frutificação em onze meses, e a prospecção fitoquímica revelou a presença de 11 classes de constituintes químicos.

ABSTRACT Phenological studies aim to determine the seasonal rhythm of the plant life cycle events, as flowering and fruiting. These events are determined by different factors, such as: alternating periods of rainy or dryer seasons, solar radiation intensity, among others. Phytochemistry aims to identify the chemical constituents of plant species or to evaluate their presence. This study aimed the phenological characterization and the phytochemical prospection of jaborandi leaves. The chosen study area for the plant material assessment was the Active Germplasm Bank of Jaborandi in the Embrapa Eastern Amazon, located in the city of Belém, PA, Brazil. The chosen accessions were the following: Merck, grown in full sun and in shade; Japanese and Bonal 4, both grown in full sun. Records were taken on a daily basis for a period of 28 months (August of 2010 to December of 2012), from five plants/accession, and arranged for monthly demonstrations by record sheets containing the corresponding plant numeration and the presence or absence of flowering and fruiting phenophases. The dry weight of the samples collected from the accessions was measured at the Laboratory of Agribusiness of Embrapa Eastern Amazon, where, after the sorting and removal of impurities, the leaves were cut, weighed, placed in stainless steel trays, and dried in forced air circulation oven (FANEM 320 UP) at a 45°C for 120h. Then, the samples were weighed, crushed, and placed in plastic bags properly identified and stored under refrigeration at a temperature of 10ºC until the use. The plant extracts were prepared using 100g of dried leaves from each accession, crushed, and subjected to hydroalcoholic extraction (80% ethanol) with water bath heating under reflux for approximately 4 hours. The extracts were stored protected from light in a refrigerator until the analysis. We analyzed the presence of the following classes of chemical substances: organic acids, reducing sugars, polysaccharides, proteins and amino acids, tannins, catechins, flavonoids, cardiac glycosides, sesquiterpene lactones, azulenes, carotenoids, steroids and triterpernoids, depsides and depsidones, coumarin derivatives, foam saponin, alkaloids, purines, anthraquinones. Our results showed that the flowering of Pilocarpus microphyllus occurred throughout the year and fruiting occurred in eleven months, and the phytochemical prospection revealed the presence of 11 classes of chemical constituents.

Cow-specific risk factors for clinical mastitis in Brazilian dairy cattle.

Information related to mastitis risk factors is useful for the design and implementation of clinical mastitis (CM) control programs. The first objective of our study was to model the risk of CM under Brazilian conditions, using cow-specific risk factors. Our second objective was to explore which risk factors were associated with the occurrence of the most common pathogens involved in Brazilian CM infections. The analyses were based on 65 months of data from 9,789 dairy cows and 12,464 CM cases. Cow-specific risk factors that could easily be measured in standard Brazilian dairy farms were used in the statistical analyses, which included logistic regression and multinomial logistic regression. The first month of lactation, high somatic cell count, rainy season and history of clinical mastitis cases were factors associated with CM for both primiparous and multiparous cows. In addition, parity and breed were also associated risk factors for multiparous cows. Of all CM cases, 54% showed positive bacteriological culturing results from which 57% were classified as environmental pathogens, with a large percentage of coliforms (35%). Coagulase-negative Staphylococcus (16%), Streptococcus uberis (9%), Streptococcus agalactiae (7%) and other Streptococci (9%) were also common pathogens. Among the pathogens analyzed, the association of cow-specific risk factors, such as Zebu breed (OR=5.84, 95%CI 3.77-10.77) and accumulated history of SCC (1.76, 95%CI 1.37-2.27), was different for CM caused by Coagulase-negative Staphylococcus and S. agalactiae in comparison to CM caused by coliforms. Our results suggest that CM control programs in Brazil should specially consider the recent history of clinical mastitis cases and the beginning of the lactations, mainly during the rainy season as important risk factor for mastitis.

Phenotypic correlations between ovum pick-up in vitro production traits and pregnancy rates in Zebu cows.

The growth of the Gyr breed in Brazil in terms of genetic gain for milk, along with conditions for market, has led to the use of ovum pick-up in vitro production (OPU-IVP) as a leader in biotechnology for the multiplication of genetic material. The aim of this study was to investigate phenotypic correlations between OPU-IVP-linked characteristics and pregnancy rates registered in an embryo transfer program using Gyr cows as oocyte donors. Data collected from 211 OPU sessions and 298 embryo transfers during the years 2012 and 2013 were analyzed and statistical analysis was performed. Estimates of simple Pearson correlations were calculated for NVcoc and PVcoc (number and proportion of viable cumulus-oocyte complexes, respectively); NcleavD4 and PcleavD4 (number and proportion of cleaved embryos on day 4 of culture, respectively); NTembD7 and PTembD7 (number and proportion of transferable embryos on day 7 of culture, respectively); NPrD30 and PPrD30 (number and proportion of pregnancies 30 days after transfer, respectively); and NPrD60 and PPrD60 (number and proportion of pregnancies 60 days after transfer, respectively). Moderate to moderately high correlations were found for all numerical characteristics, suggesting these as the most suitable parameters for selection of oocyte donors in Gyr programs. NVcoc is proposed as a selection trait due to positive correlations with percentage traits and pregnancy rates 30 and 60 days after transfer.

Cathepsin D protects colorectal cancer cells from acetate-induced apoptosis through autophagy-independent degradation of damaged mitochondria.

Acetate is a short-chain fatty acid secreted by Propionibacteria from the human intestine, known to induce mitochondrial apoptotic death in colorectal cancer (CRC) cells. We previously established that acetate also induces lysosome membrane permeabilization in CRC cells, associated with release of the lysosomal protease cathepsin D (CatD), which has a well-established role in the mitochondrial apoptotic cascade. Unexpectedly, we showed that CatD has an antiapoptotic role in this process, as pepstatin A (a CatD inhibitor) increased acetate-induced apoptosis. These results mimicked our previous data in the yeast system showing that acetic acid activates a mitochondria-dependent apoptosis process associated with vacuolar membrane permeabilization and release of the vacuolar protease Pep4p, ortholog of mammalian CatD. Indeed, this protease was required for cell survival in a manner dependent on its catalytic activity and for efficient mitochondrial degradation independently of autophagy. In this study, we therefore assessed the role of CatD in acetate-induced mitochondrial alterations. We found that, similar to acetic acid in yeast, acetate-induced apoptosis is not associated with autophagy induction in CRC cells. Moreover, inhibition of CatD with small interfering RNA or pepstatin A enhanced apoptosis associated with higher mitochondrial dysfunction and increased mitochondrial mass. This effect seems to be specific, as inhibition of CatB and CatL with E-64d had no effect, nor were these proteases significantly released to the cytosol during acetate-induced apoptosis. Using yeast cells, we further show that the role of Pep4p in mitochondrial degradation depends on its protease activity and is complemented by CatD, indicating that this mechanism is conserved. In summary, the clues provided by the yeast model unveiled a novel CatD function in the degradation of damaged mitochondria when autophagy is impaired, which protects CRC cells from acetate-induced apoptosis. CatD inhibitors could therefore enhance acetate-mediated cancer cell death, presenting a novel strategy for prevention or therapy of CRC.

Yeast as a tool to explore cathepsin D function.

Cathepsin D has garnered increased attention in recent years, mainly since it has been associated with several human pathologies. In particular, cathepsin D is often overexpressed and hypersecreted in cancer cells, implying it may constitute a therapeutic target. However, cathepsin D can have both anti- and pro-survival functions depending on its proteolytic activity, cellular context and stress stimulus. Therefore, a more detailed understanding of cathepsin D regulation and how to modulate its apoptotic functions is clearly needed. In this review, we provide an overview of the role of cathepsin D in physiological and pathological scenarios. We then focus on the opposing functions of cathepsin D in apoptosis, particularly relevant in cancer research. Emphasis is given to the role of the yeast protease Pep4p, the vacuolar counterpart of cathepsin D, in life and death. Finally, we discuss how insights from yeast cathepsin D and its role in regulated cell death can unveil novel functions of mammalian cathepsin D in apoptosis and cancer.

Preventive effect of CuCl2 on behavioral alterations and mercury accumulation in central nervous system induced by HgCl2 in newborn rats.

This study investigated the benefits of Cu preexposition on Hg effects on behavioral tests, acetylcholinesterase (AChE) activity and Hg, and essential metal contents in the cerebrum and cerebellum of neonate rats. Wistar rats received (subcutaneous) saline or CuCl2 ·2H2O (6.9 mg/kg/day) when they were 3 to 7 days old and saline or HgCl2 (5.0 mg/kg/day) when they were 8 to 12 days old. Mercury exposure reduced the performance of rats in the negative geotaxis (3-13 days) and beaker test (17-20 days), inhibited cerebellum AChE activity (13 days), increased cerebrum and cerebellum Hg (13 days), cerebrum Cu (13 days), and cerebrum and cerebellum Zn levels (33 days). The performance of rats in the tail immersion and rotarod tests as well as Fe and Mg levels were not altered by treatments. Copper prevented all alterations induced by mercury. These results are important to open a new perspective of prevention and/or therapy for mercury exposure.