Total rupture of Achilles tendon induces inflammatory response and glial activation on the spinal cord of mice

Rupture of Achilles tendon is a common accident affecting professional and recreational athletes. Acute and chronic pain are symptoms commonly observed in patients with rupture. However, few studies have investigated whether Achilles tendon rupture is able to promote disorders in the central nervous system (CNS). Therefore, the current study aimed to evaluate nociceptive alterations and inflammatory response in the L5 lumbar segment of Balb/c mice spinal cord after Achilles tendon rupture. We found increased algesia in the paw of the ruptured group on the 7th and 14th days post-tenotomy compared with the control group. This phenomenon was accompanied by overexpression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase-2 (NOS-2) as well as hyperactivation of astrocytes and microglia in nociceptive areas of L5 spinal cord as evidenced by intense GFAP and IBA-1 immunostaining, respectively. Biochemical studies also demonstrated increased levels of nitrite in the L5 spinal cord of tenotomized animals compared with the control group. Thus, we have demonstrated for the first time that total rupture of the Achilles tendon induced inflammatory response and nitrergic and glial activation in the CNS in the L5 spinal cord region.

P2X7 receptor antagonist improves gastrointestinal disorders in spontaneously hypertensive rats

The purinergic system participates in the control of blood pressure. Hypertension promotes the occurrence of gastrointestinal disorders such as intestinal inflammation and gastric emptying delay. This study aimed i) to investigate the participation of the P2X7 receptor blocker Brilliant Blue G (BBG) on gastric emptying of solids and changes in oxidative stress in the gastric fundus, duodenum, and colon of spontaneously hypertensive rats (SHR) and ii) to study the putative relationship of this effect with the renin-angiotensin system. Rats were divided into five groups: Control, SHR, SHR+BBG, SHR+BBG+ATP, and SHR+BBG+ANG II. In the gastrointestinal tract, we assessed gastric emptying (GE) and oxidative stress markers (NOx, MPO, GSH, SOD). We observed a decrease in the GE rate (P<0.05) in SHR vs control rats (21.8±2.0% vs 42.8±3.5%). The decrease in GE was returned (P<0.05) to control levels by BBG in SHR rats (21.8±2.0% vs 41.6±3.2%). Co-administration of ATP or ANG II together with BBG bypassed the effect of the P2X7 antagonist on GE in SHR (P<0.05) (21.9±5.0% vs 25.6±3.0% vs 41.6±3.2%). The MPO activity increased (P<0.05) in the gastric fundus of SHR compared to control rats (6.12±2.26 vs 0.077±0.02 UMPO/mg tissue); this effect was prevented (P<0.05) by BBG (0.55±0.15 vs 6.12±2.26 UMPO/mg tissue). Data demonstrated that blockage of P2X7 receptors with BBG can improve the GE delay and oxidative stress biomarkers in SHR animals. This preventive effect of BBG on GE delay was abrogated by ANG II and ATP, thus prompting crosstalk between renin-angiotensin and the purinergic signaling systems underlying this phenomenon.

Fructose consumption induces molecular adaptations involving thyroid function and thyroid-related genes in brown adipose tissue in rats

The increasing incidence of metabolic diseases is in part due to the high fructose consumption, a carbohydrate vastly used in industry, with a potent lipogenic capacity. Thyroid hormones (TH) are essential for metabolism regulation and are associated with changes in body weight, energy expenditure, insulin sensitivity, and dyslipidemia. This study aimed to investigate the influence of fructose intake on thyroid function and thyroid-related genes. Male Wistar rats were divided into Control (CT, n=8) and Fructose (FT - 10% in drinking water, n=8) groups for three weeks. The FT group showed higher glycemia and serum triacylglycerol, indicating metabolic disturbances, and increased thyroid mass, accompanied by higher expression of Srebf1c and Lpl, suggesting increased lipid synthesis. The FT group also presented higher expression of Tpo and Dio1 in the thyroid, suggesting activation of the thyroid gland, but with no alterations in serum TH concentrations. Brown adipose tissue (BAT) of the FT group exhibited higher expression of Dio2, Thra, and Thrb, indicating increased T3 intra-tissue bioavailability and signaling. These responses were accompanied by increased BAT mass and higher expression of Adrb3, Pparg, Srebf1c, Fasn, Ppara, and Ucp1, suggesting increased BAT adrenergic sensitivity, lipid synthesis, oxidation, and thermogenesis. Therefore, short-term fructose consumption induced thyroid molecular alterations and increased BAT expression of thyroid hormone-related signaling genes that potentially contributed to higher BAT activity.

Carnauba (Copernicia prunifera) straw as an alternative bedding material for dairy cows housed in a Compost barn system / [Bagana de carnaúba como material de cama alternativo para vacas leiteiras alojadas em sistema Compost barn]

The replacement of bedding in compost dairy barns (CB) comprises a recurrent management practice, but bedding materials are often not readily available in all regions and the choice of alternative materials is necessary. The objective was to evaluate the thermal attributes of carnauba straw (CS) bedding in compost dairy barn facilities. Environmental monitoring operations were performed at a commercial farm located in Northeast Brazil. Mini weather stations were used to evaluate environmental variables. The THI was evaluated as one of the comfort parameters. Analysis of the spatial distribution of bed surface temperature (BST) in the CB was performed using geostatistical techniques. The cows remained out of the comfort zone according to THI results. The BST indicated satisfactory performance and from the thermal point of view can be used as alternative bedding material in CB facilities. However, it was observed that the CS showed fast biomass degradation compared to conventional materials, widely known. In addition, inadequate temperature values (< 45°C) were found in the deeper of the CS bed, signaling higher risks of pathogenic microbial activity. Additional studies are needed for searching the proper management plans that increase the life span of the bed formed by carnauba straw.(AU)

A reposição de cama na instalação Compost Barn (CB) compreende uma prática recorrente de manejo, porém os materiais de cama muitas vezes não estão facilmente disponíveis em todas as regiões, sendo necessária a adoção de materiais alternativos. Objetivou-se avaliar os atributos térmicos da cama à base de bagana de carnaúba (BC) para instalações CB. Foram realizadas operações de monitoramento ambiental em uma fazenda comercial situada no nordeste do Brasil. Miniestações meteorológicas foram utilizadas para avaliação de variáveis ambientais. O ITU foi avaliado como um dos parâmetros de conforto. A distribuição espacial da temperatura superficial da cama (TSC) no galpão foi analisada usando técnicas da geoestatística. As vacas mantiveram-se fora da zona de conforto, conforme resultados do ITU. A TSC indicou desempenho satisfatório e, do ponto de vista térmico, pode ser utilizada como material alternativo para cama em instalações CB. Entretanto, observou-se que a BC apresentou rápida degradação da biomassa em comparação a materiais convencionais, amplamente conhecidos. Além disso, valores inadequados de temperatura (< 45°C) foram encontrados nas camadas profundas da cama de BC, sinalizando maiores riscos de atividade microbiana patogênica. Estudos adicionais tornam-se necessários para a busca de planos de manejo que aumentem o tempo de vida útil da cama formada por bagana de carnaúba.(AU)

The essential oil of Hyptis crenata Pohl ex Benth. presents an antiedematogenic effect in mice

Hyptis crenata, commonly known as "salva-do-Marajó", "hortelã-do-campo", and "hortelãzinha", is used in folk medicine in Northeast Brazil as tea or infusion to treat inflammatory diseases. Due to the pharmacological efficacy and the low toxicity of the essential oil of Hyptis crenata (EOHc), we decided to investigate the EOHc antiedematogenic effect in experimental models of inflammation. EOHc was administrated orally at doses of 10-300 mg/kg to male Swiss albino mice. Paw edema was induced by subcutaneous injection in the right hind paw of inflammatory stimuli (carrageenan, dextran, histamine, serotonin, and bradykinin) 60 min after administration of EOHc. EOHc significantly inhibited the induced edema. The inhibitory effect of EOHc on dextran-induced edema extended throughout the experimental time. For the 30, 100, and 300 mg/kg doses of EOHc, the inhibition was of 40.28±1.70, 51.18±2.69, and 59.24±2.13%, respectively. The EOHc inhibitory effect on carrageenan-induced edema started at 10 mg/kg at the second hour (h) and was maintained throughout the observation period. At 30, 100, and 300 mg/kg doses the inhibition started earlier, from 30 min. At the edema peak of 180 min, 56, 76, and 82% inhibition was observed for 30, 100, and 300 mg/kg doses, respectively. Additionally, the effect of EOHc on carrageenan-induced paw edema was influenced by the time of administration. The EOHc also inhibited myeloperoxidase activity. In conclusion, the EOHc showed a potent effect, both preventing and reversing the edema, consistent with its anti-inflammatory use in folk medicine.

Avaliação nutricional e metabólica de borregas alimentadas com níveis crescentes de aminoácidos protegidos / [Nutritional and metabolic evaluation of feeding lambs with growing levels of protein amino acids]

Objetivou-se avaliar a inclusão de níveis de lisina e metionina protegidas na dieta sobre os parâmetros nutricionais e metabólicos energéticos e hepáticos de borregas em crescimento. Utilizaram-se cinco borregas ½ sangue Dorper x Santa Inês, com aproximadamente oito meses de idade e peso médio de 50 ± 2,3kg, distribuídas em esquema quadrado latino 5x5 (cinco tratamentos, cinco animais e cinco períodos). Os tratamentos consistiram na inclusão de diferentes níveis de lisina e metionina protegidas da degradação ruminal (MicroPEARLS LM®) na ração, sendo: 0g, 8g, 16g, 24g e 32g por dia. A dieta era composta por silagem de milho e concentrado na relação 30V:70C. Realizou-se um ensaio de digestibilidade para determinar consumo e digestibilidade da matéria seca (CMS/DGMS), balanço de nitrogênio e metabólitos sanguíneos. O CMS (kg/dia) em relação ao peso metabólico apresentou equação linear positiva, sendo maior no tratamento que ofertou 32g de aminoácidos por dia, assim como o nitrogênio ingerido e o balanço de nitrogênio, sendo positivo em todos os tratamentos. Não houve diferença (P>0,05) para a digestibilidade da MS e o metabolismo energético e hepático. Lisina e metionina protegidas da degradação ruminal podem ser incluídas na ração de borregas em crescimento até 32g/dia sem causar efeitos negativos na digestibilidade da MS e no metabolismo.(AU)

The objective was to evaluate the inclusion of protected lysine and methionine levels on the diet, over the nutritional parameters and energetic and hepatic metabolites of growing lambs. Five lambs ½ blood Dorper x Santa Inês, with approximately eight months of age and average weight of 50kg, were distributed in a 5x5 latin square scheme (five treatments and five replicates). The treatments consisted of the inclusion of different levels of lysine and methionine protected from ruminal degradation (MicroPEARLS LM®) in the diet, being: 0g, 8g, 16g, 24g and 32g. The diet was composed of corn silage and concentrated 30V:70C in the ratio. A digestibility assay was performed to determine dry matter intake and digestibility (DMI/DDMI), nitrogen balance and blood metabolites. The DMI (kg/day) in relation to the metabolic weight had a positive linear equation, being higher in treatment 32g, as well as the ingested nitrogen and nitrogen balance, being positive in all treatments. There was no difference (P>0.05) for the digestibility of DM, energetic and hepatic metabolism. Lysine and methionine protected from ruminal degradation can be included in the diet of growing lambs up to 32g without causing negative effects on DM digestibility and metabolism.(AU)

Efeito do hipotireoidismo materno na expressão espaço-temporal de mediadores imunológicos e população de células natural killers na decídua e na glândula metrial de ratas / Effect of maternal hypothyroidism on the spatiotemporal expression of immunological mediators and population of Natural Killers cells in decidua and metrial gland of rats

Estudou-se o efeito do hipotireoidismo materno na expressão espaço-temporal de mediadores imunológicos e na população de células natural killers (NK) na decídua e na glândula metrial de ratas durante a gestação. Avaliou-se a detecção imunoistoquímica de interferon γ (IFNγ), do fator inibidor de migração (MIF), da interleucina 15 (IL15), do óxido nítrico sintase induzível (iNOS), a marcação com lectina DBA para evidenciação das células NK uterinas DBA+ e a expressão gênica de Ifnγ e Nos2. O hipotireoidismo aumentou o iNOS aos sete dias, a IL15 e o MIF aos 10 e 12 dias, o IFNγ e o MIF aos 14 DG e a expressão dos transcritos gênicos para iNos aos 12 e 19 dias e para Ifnγ aos 14 DG. O hipotireoidismo reduziu a imunomarcação de MIF e lectina DBA aos sete dias, lectina DBA aos 10 e 14 DG, IFNγ aos 12 dias, e a expressão de Ifnγ aos 10 e 19 DG e de iNOS aos 12, 14 e 19 DG, bem como reduziu seus transcritos gênicos aos 10 e 14 DG. Conclui-se que o hipotireoidismo compromete o perfil imunológico na interface materno-fetal ao longo da gestação, particularmente por reduzir o fator anti-inflamatório iNOS e a população de células uNK DBA+.(AU)

The goal of this study was to evaluate the effect of maternal hypothyroidism on the spatiotemporal expression of immunological mediators and population of Natural Killers cells in decidua and metrial gland of rats. Interferon gamma (IFNγ), migration inhibitory factor (MIF), interleukin 15 (IL15), inducible nitric oxide synthase (iNOS), and DBA-Lectin labeling for evidence of uNK DBA+ cells in decidua and genetic expression of Ifnγ and iNos by real-time RT-PCR were evaluated. Hypothyroidism increased protein expression of iNOS at 7 days, IL15 and MIF at 10 and 12 days, IFNγ and MIF at 14 DG in the decidua and/or metrial gland and the gene transcripts for iNOS at 12 and 19 days and for Inf at 14 DG. In addition, hypothyroidism reduced the protein expression of MIF and DBA-Lectin at 7 days, DBA-Lectin at 10 and 14 DG, IFNγ at 12 days, and the gene transcript to Ifnγ at 10 and 19 DGs. Hypothyroidism also reduced the protein expression of iNOS at 12, 14 and 19 DG and reduced its gene transcripts at 10 and 14 DGs. It is concluded that hypothyroidism compromises the immunology profile at the maternal-fetal interface throughout pregnancy, particularly by reducing the anti-inflammatory factor iNOS and population of uNK DBA+ cells.(AU)

Direct inhibitory effect of caffeine on viability, synthesis activity and gene expression in cultures of chondrocytes extracted from the articular cartilage of rats / Efeito inibidor direto da cafeína sobre a viabilidade, atividade de síntese e expressão gênica em culturas de condrócitos da cartilagem articular de ratos

The aim of this study was to evaluate the effect of concentrations of caffeine on the viability, synthesis activity and gene expression in cultures of chondrocytes. Extracted articular cartilage from the femurs and tibias of 15 Wistar rats at three days old to isolate chondrocytes. Chondrocytes were cultured in chondrogenic medium (control) or supplemented with caffeine (0.5, 1.0, 2.0mM). Cell viability, alkaline phosphatase activity and collagen synthesis were assessed using colorimetric assays at 7, 14, 21 days. The chondrocyte cultures of all groups grown under coverslips were stained with hematoxylin-eosin to determine the percentage of cells/field and with PAS, safranin O, alcian blue to determine the percentage of matrix chondrogenic/field at 21 days. The expressions of gene transcripts for aggrecan, collagen-II, Sox-9, Runx-2 and alkaline phosphatase were also evaluated by RT-PCR at 21 days. The means were compared using Student-Newman-Keuls. Caffeine significantly reduced the conversion of MTT to formazan, percentage of cells/field, collagen synthesis, alkaline phosphatase activity, synthesis of PAS+, safranin O+ and alcian blue+ chondrogenic matrix, and the expression of aggrecan, Sox-9 and II collagen. It is concluded that caffeine at concentrations of 0.5, 1.0, 2.0mM has a direct inhibitory effect on chondrogenesis in cultures of chondrocytes from rats.(AU)

O objetivo deste estudo foi avaliar o efeito direto de concentrações de cafeína sobre a viabilidade, atividade de síntese e expressão gênica em culturas de condrócitos de ratos. As cartilagens dos fêmures e tíbias de 15 ratos Wistar com três dias foram extraídas para isolamento de condrócitos. Os condrócitos foram cultivados em meio condrogênico (controle) ou em meio acrescido de diferentes concentrações de cafeína (0,5, 1,0, 2,0mM). Foram avaliadas a viabilidade celular, a atividade da fosfatase alcalina e a síntese de colágeno por ensaios colorimétricos aos sete, 14 e 21 dias. Condrócitos cultivados sob lamínulas foram corados pela hematoxilina e eosina, para se determinar a porcentagem de células/campo, e pelo PAS, safranina O, alcian Blue, para se determinar a porcentagem de matriz condrogênica/campo aos 21 dias. Foi avaliada a expressão de transcriptos gênicos para Sox-9, Runx-2, agrecano, colágeno-II e fosfatase alcalina por qRT-PCR, aos 21 dias. As médias foram comparadas pelo Student-Newman-Keuls. A cafeína reduziu significativamente o MTT em cristais de formazan, a porcentagem de células/campo, a síntese de colágeno, a atividade da fosfatase alcalina e a síntese de matriz condrogênica PAS+, safranina O+, alcian blue+ e expressão de Sox-9 e colágeno-II. Conclui-se que a cafeína, nas concentrações de 0,5, 1,0, 2,0mM, apresenta efeito inibidor direto sobre a condrogênese em culturas de condrócitos de ratos.(AU)

Local treatment with ascorbic acid accelerates recovery of post-sutured Achilles tendon in male Wistar rats

Tendon rupture is a very frequent accident involving average people and high-performance athletes. Clinical studies describe tendon recovery as a painful and slow process involving different biochemical and histological events. Ascorbic acid (AA) is a potent antioxidant as well as an important cofactor for collagen synthesis. In the current study, we evaluated if local treatment with AA is able to promote tendon repair in tenotomized rats. Animals were submitted to Achilles tendon rupture followed by surgical suture. Control and AA groups received in loco injection of saline solution (0.9% NaCl) and 30 mM AA, respectively. Histological and functional recovery of Achilles tendon tissue was evaluated at 7, 14, and 21 days post-surgery. Hematoxylin/eosin staining and collagen fluorescence analysis showed intense disarrangement of tendon tissue in the saline group. Tenotomized animals also showed hypercellularity in tendon tissue compared with non-tenotomized animals. The Achilles functional index (AFI) showed a significant decrease of tendon functionality in tenotomized animals at 7, 14, and 21 days post-surgery. AA accelerated tissue organization and the recovery of function of the Achilles tendons. The beneficial effect of AA treatment was also observed in the organization of the collagen network. Data presented in the current work showed that in loco treatment with AA accelerated the recovery of injured Achilles tendon post-surgery.

Essential oil of Pterodon polygalaeflorus Benth attenuates nociception in mice

Essential oils (EO) are volatile liquids responsible for the aroma of plants. Pterodon polygalaeflorus seeds have received widespread use in folk medicine for the treatment of inflammatory diseases. For this reason and because Pterodon polygalaeflorus seeds have great EO content, which is frequently pharmacologically active, the present study aimed to evaluate the antinociceptive effect of EO from Pterodon polygalaeflorus (EOPPgfl) and its acute toxic effects. The EEOPPgfl sample, which was extracted by steam distillation of the seeds, had a yield of 2.4% of the seeds weight and had, as major constituents, beta-elemene (48.19%), trans-caryophyllene (19.51%), and epi-bicyclosesquiphellandrene (12.24%). The EOPPgfl sample showed mild acute toxicity and its calculated median lethal dose (LD50) was 3.38 g/kg. EOPPgfl (20-60 mg/kg) showed antinociceptive activity as evidenced by several tests and inhibited writhing induced by acetic acid. The maximum effect was obtained with the 30 mg/kg dose and at 60 min after its administration. EOPPgfl also decreased formalin-induced nociception, as verified by the inhibition of the first and second phase of the formalin test. At 30 mg/kg, EOPPgfl also decreased thermally stimulated nociception. Nociception may be related to inflammatory and antiedematogenic activity and at doses ranging 10-100 mg/kg, EOPPgfl blocked dextran- and carrageenan-induced edema. The results demonstrated that EOPPgfl presented, at doses approximately 100 times smaller than LD50, an antinociceptive effect that probably was due to anti-inflammatory activities.

Benefícios da condroitinase abc associada a células-tronco mesenquimais na lesão espinhal aguda em ratos / Benefits of chondroitinase abc associated with mesenchymal stem cells in acute spinal cord injury in rats

Com o objetivo de estudar o efeito da condroitinase associada às células-tronco mesenquimais na lesão aguda da medula espinhal, utilizaram-se 50 ratos Lewis, distribuídos igualmente nos grupos: controle negativo (CN), tratamento com placebo (PLA), condroitinase (CDN), células-tronco mesenquimais (CTM) e condroitinase mais células-tronco mesenquimais (CDN+CTM). Todos os animais tiveram a medula espinhal exposta por laminectomia, e os grupos PLA, CDT, CTM e CDT+CTM sofreram também trauma medular compressivo. Após sete dias, procedeu-se à reexposição da medula espinhal, quando os grupos PLA e CTM receberam 4µL de líquido cefalorraquidiano artificial via intralesional, e os grupos CDT e CDT+CTM receberam o mesmo líquido contendo 2,2U de condroitinase. Após 14 dias da cirurgia inicial, todos os animais receberam 0,2mL de PBS via endovenosa, contudo, nos grupos CTM e CDT+CTM, esse líquido continha 1x106 CTM. Avaliou-se a capacidade motora até o 28o dia pós-trauma e, posteriormente, as medulas espinhais foram analisadas por RT-PCR, para quantificação da expressão gênica para BDNF, NT-3, VEGF, KDR e PECAM-1, e por imunoistoquímica, para detecção das células-tronco GFP injetadas (anti-GFP), quantificação dos neurônios (anti-NeuN) e da GFAP e vimentina, para avaliação da cicatriz glial. As análises estatísticas foram realizadas com o auxílio do Prism 5 for Windows, com o nível de significância de 5%. Não houve diferença entre os grupos quanto à capacidade motora. O grupo CDT+CTM apresentou maior imunoexpressão de neurônios viáveis do que o placebo. No CTM, houve maior expressão dos fatores neurotróficos BDNF e VEGF. E no CDT, houve menor imunoexpressão de vimentina. Concluiu-se que a associação CDT+CTM favorece a viabilidade neuronal após o trauma, que o tratamento com CTM promove aumento na expressão dos fatores tróficos BDNF e VEGF e que o tratamento com condroitinase é efetivo na redução da cicatriz glial.(AU)

The aim of this work was to study the effect of chondroitinase associated with mesenchymal stem cells in acute spinal cord injury. Therefore, 50 Lewis rats were distributed in the following groups: negative control (NC), treatment with placebo (PLA), chondroitinase (CDT), mesenchymal stem cells (MSC), and chondroitinase associated with mesenchymal stem cells (CDT + MSC). All animals had their spinal cord exposed by laminectomy, and the groups named PLA, CDT, MSC and CDT + MSC also suffered compressive spinal cord trauma. After seven days, the spinal cord was re-exposed, when the PLA and MSCs groups received 4uL of artificial cerebrospinal fluid through the lesion, and the CDT group and CDT + MSC received the same fluid containing 2,2U of chondroitinase. 14 days after the first surgery, all animals received 0.2ml of PBS intravenously; however, the MSC and CDT + MSC groups received the same liquid also containing 1x106 MSCs. The motor skills were evaluated up to 28 days post-injury and, subsequently, the spinal cords were analyzed by RT-PCR for BDNF, NT-3, VEGF, PECAM-1 and KDR gene expression quantification, immunohistochemistry to detect injected stem cells GFP (anti-GFP), to quantify neurons (anti-NeuN), GFAP and detect vimentin in order to evaluate the glial scar. Statistical analyzes were performed by Prism 5 for Windows using a 5% level of significance. There was no difference between groups with regarding motor capacity. The CDT + MSC group showed increased immunoreactivity of viable neurons than placebo. In MSC, there was a greater expression of neurotrophic factors BDNF and VEGF. Also, there was less vimentin immunostaining in group CDT. It was concluded that CDT + MSC association promotes neuronal viability after trauma, in which treatment with MSC promotes increased expression of BDNF and VEGF trophic factors, and also that treatment with chondroitinase is effective in reducing the glial scar.(AU)

Osteogenic differentiation of adipose tissue-derived mesenchymal stem cells cultured with different concentrations of prolactin / Diferenciação osteogênica de células tronco mesenquimais do tecido adiposo cultivadas com diferentes concentrações de prolactina

The objective was to evaluate the in vitro effect of prolactin in osteogenic potential of adipose tissue-derived mesenchymal stem cells (ADSCs) in female rats. ADSCs were cultured in osteogenic medium with and without the addition of prolactin and distributed into three groups: 1) ADSCs (control), 2) ADSCs with addition of 100ng/mL of prolactin and 3) ADSCs with addition of 300ng/mL of prolactin. At 21 days of differentiation, the tests of MTT conversion into formazan crystals, percentage of mineralized nodules and cells per field and quantification of genic transcript for alkaline phosphatase, osteopontin, osteocalcin, bone sialoprotein, BMP-2 and collagen I by real-time RT-PCR were made. The addition of prolactin reduced the conversion of MTT in group 3 and increased the percentage of cells per field in the groups 2 and 3, however without significantly increasing the percentage of mineralized nodules and the expression of alkaline phosphatase, osteopontin, osteocalcin, bone sialoprotein, BMP-2 and collagen I. In conclusion, the addition of prolactin in concentrations of 100ng/mL and 300ng/mL does not change the osteogenic differentiation to the ADSCs of female rats despite increase in the cellularity of the culture.(AU)

O objetivo do presente trabalho foi avaliar o efeito in vitro da prolactina sobre o potencial osteogênico de células-tronco mesenquimais do tecido adiposo (CTM-TA) em ratas. CTM-TA foram cultivadas em meio osteogênico com e sem adição de prolactina e distribuídas em três grupos: 1) CTM-TA (controle), 2) CM-TA com adição de 100ng/mL de prolactina e 3) CTM-TA com adição de 300ng/mL de prolactina. Aos 21 dias de diferenciação, foram realizados os testes de conversão do MTT em cristais de formazan, porcentagem de nódulos mineralizados e células por campo e quantificação dos transcritos gênicos para fosfatase alcalina, osteopontina, osteocalcina, sialoproteína óssea, BMP-2 e colágeno I. A adição de prolactina reduziu a conversão do MTT no grupo 3 e aumentou a porcentagem de células por campo nos grupos 2 e 3, sem alterar significativamente a porcentagem de nódulos mineralizados e a expressão de fosfatase alcalina, osteopontina, osteocalcina, sialoproteína óssea, BMP-2 e colágeno I. Conclui-se que a adição de prolactina nas concentrações de 100ng/mL e 300ng/mL não altera a diferenciação osteogênica das CTM-TA de ratas, apesar do aumento de celularidade da cultura.(AU)

Comportamento ingestivo de bovinos leiteiros alimentados com farelo de crambe / Ingestive behavior of dairy catle fed crambe bran

O objetivo deste trabalho foi estudar os efeitos da utilização de farelo de crambe em substituição ao farelo de soja sobre o comportamento ingestivo de bovinos leiteiros. Foram utilizados quatro machos castrados Holandês x Zebu, fistulados no rúmen, com peso vivo médio de 664kg, distribuídos em delineamento quadrado latino 4 x 4. Os tratamentos consistiram em quatro dietas isoproteicas e isoenergéticas, formuladas com relação volumoso:concentrado 60:40 com base na matéria seca (MS). O volumoso foi composto de silagem de milho (51% MS) e feno de Tifton (49% MS), e o concentrado formulado com níveis crescentes de substituição do farelo de soja pelo farelo de crambe em 0%, 2,8%, 6,4% e 11,0% na MS da dieta. O comportamento ingestivo foi avaliado por meio do método direto de avaliação visual, em intervalos de 10 minutos, durante períodos de 24 horas. Registraram-se a frequência de alimentação, a ruminação e o ócio, bem como a posição do animal (em pé ou em decúbito). As variáveis em pé e em decúbito não diferiram entre os tratamentos, assim como os tempos gastos em alimentação, ruminação e ócio. O consumo de matéria seca e de FDN expressos em g/dia e gFDN/dia, respectivamente, a eficiência de ruminação expressa em gMS/min, a eficiência de ruminação expressa em gFDN/dia e o tempo de mastigação total não diferiram significativamente. No entanto, a eficiência de alimentação (gMS/min) variou de forma linear decrescente com a inclusão do farelo de crambe. Os períodos do dia influenciaram todas as atividades. O maior tempo de alimentação foi observado nos períodos após o fornecimento da dieta, e a maior atividade de ruminação foi verificada no período noturno. A substituição de farelo de soja por farelo de crambe não afetou o comportamento ingestivo, exceto para o parâmetro eficiência de alimentação. Nesse sentido, considerando o comportamento ingestivo, recomenda-se a substituição do farelo de soja por farelo de crambe para alimentação de bovinos leiteiros.

The aim of this work was to study the effects of the use of crambe meal replacing soy bean meal on the ingestive behavior of dairy cattle. Four male Holstein x Zebu rumen, with average live weight of 664 kg distributed in a 4x4 latin square design. The treatments consisted of four isonitrogenous and isocaloric diets, formulated with a roughage:concentrate ratio of 60:40 based on dry matter (DM). The forage was corn silage (51% DM) and Tifton grass hay (49% DM), and the concentrate was formulated with increasing levels of substitution of soy bean meal by crambe meal at 0%, 2.8%, 6.4% and 11.0% of the diet DM. The feeding behavior was evaluated using the direct method of visual assessment in 10 minute intervals during a 24 hour period. The frequency of feeding, idling time and the animal's position (standing or supine) were recorded. The variables standing and supine did not differ between treatments as well as time spent feeding, ruminating and idling times. The consumption of dry matter and NDF expressed in g/day and gNDF/day respectively, the efficiency of rumination expressed in gDM/min, the efficiency of rumination expressed in gNDF/day and the total chewing time did not differ significantly. However, the efficiency of power (gDM/min) decreased linearly with the inclusion of crambe meal. Periods of the day influenced all the activities. The longer feeding time was observed in the periods after the diet intake and greater rumination activity was observed at night. The replacement of soy bean meal by meal crambe did not affect feeding behavior, except for the feeding efficiency parameter. In this sense, considering the feeding behavior, the replacement of soy bean meal by crambe meal to feed dairy cattle is recommended.

Electroretinography and immunohistochemistry of retina in rabbits treated with sildenafil citrate / Eletrorretinografia e imuno-histoquímica da retina de coelhos tratados com citrato de sildenafil

Sildenafil citrate is a type-5 phosphodiesterase inhibitor (PDE-5), able to inhibit type-6 phosphodiesterase (PDE-6) as well, providing clinical benefits and paraeffects, some of them potentially related to the retina. The effects of the sildenafil on the retrobulbar and retinal circulation were studied in 27 adult male rabbits of the White New Zealand breed. The electric activity of the retina was evaluated before and at the end of the treatments, and immunohistochemistry studies were conducted. An amplitude increase of the b wave was found in the mixed response of cones and rods after 7 days of treatment with sildenafil citrate. However, in the other evaluations and periods, the values did not differ from the basal ones. Through immunohistochemistry, no significant decrease of the expression of PDE-5 and PDE-6 proteins was observed. Based on the results obtained, it is possible to admit that the sildenafil citrate did not change the expression of PDE-5 and PDE-6, neither the electroretinographic activity of the retina of male rabbits of the White New Zealand breed.

O citrato de sildenafil é um inibidor da fosfodiesterase do tipo 5 (PDE-5), capaz de inibir também a fosfodiesterase do tipo 6 (PDE-6), proporcionando benefícios clínicos e paraefeitos, alguns deles potencialmente relacionados à retina. Foram estudados efeitos do sildenafil sobre a circulação retrobulbar e a retiniana em 27 coelhos machos adultos, da raça Nova Zelândia Branco. Avaliou-se a atividade elétrica da retina antes e ao término dos tratamentos e realizaram-se estudos à imunoistoquímica. Encontrou-se aumento da amplitude da onda b na resposta mista de cones e de bastonetes, após 7 dias de tratamento com citrato de sildenafil. Entretanto, nas demais avaliações e períodos, os valores não divergiram dos basais. Pela imunoistoquímica, não se observou diminuição significativa da expressão das proteínas PDE-5 e PDE-6. Com base nos resultados obtidos, é possível admitir que o citrato de sildenafil não alterou a expressão de PDE-5 e PDE-6, tampouco, a atividade eletrorretinográfica da retina de coelhos machos da raça Nova Zelândia Branco.

Cultivation of marine shrimp in biofloc technology (BFT) system under different water alkalinities / Cultivo de camarões marinhos em sistema de bioflocos (BFT) sob diferentes alcalinidades da água

AbstractThe aim of this study was to evaluate the influence of different levels of alkalinity for the superintensive cultivation of marine shrimp Litopenaeus vannamei in biofloc system. A total of 12 experimental circular units of 1000L were used supplied with 850L water from a nursery, populated at a density of 165 shrimps.m–3 and average weight of 5.6 g. The treatments, in triplicate, consisted in four levels of alkalinity in the water: 40, 80, 120 and 160 mg.L–1 of calcium carbonate. To correct the alkalinity was used calcium hydroxide (CaOH). It was observed a decrease in pH of the water in the treatments with lower alkalinity (p<0.05). The total suspended settleable solids were also lower in the treatment of low alkalinity. No significant difference was observed in other physico-chemical and biological parameters in the water quality assessed, as well as the zootechnical parameters of cultivation between treatments (p≥0.05). The results of survival and growth rate of shrimps were considered suitable for the cultivation system used in the different treatments. The cultivation of marine shrimp Litopenaeus vannamei in biofloc at density of 165 shrimps.m–3 can be performed in waters with alkalinity between 40 and 160 mg.L–1 of CaCO3, without compromising the zootechnical indexes of cultivation.

ResumoO objetivo deste trabalho foi avaliar a influência de diferentes níveis de alcalinidade para o cultivo do camarão marinho Litopenaeus vannamei em sistema superintensivo em bioflocos. Foram utilizadas 12 unidades experimentais circulares de 1000L abastecido com 850L de água provenientes de um berçário intensivo, povoadas a uma densidade de 165 camarões.m-3 e peso médio 5,6g. Os tratamentos em triplicata consistiram de quatro níveis de alcalinidade na água: 40, 80, 120 e 160 mg.L–1 de carbonato de cálcio. Para correção da alcalinidade, foi utilizado cal hidratada (CaOH). Foi observado um decréscimo no pH da água nos tratamentos de menor alcalinidade (p<0,05). Os sólidos suspensos sedimentáveis totais também foram menores nos tratamentos de menor alcalinidade. Não foi observada diferença significativa nos demais parâmetros físico-químicos e biológicos de qualidade de água avaliados, assim como nos parâmetros zootécnicos do cultivo entre os tratamentos (p≥0,05). Os resultados de sobrevivência e taxa de crescimento dos camarões foram considerados adequados para o sistema de cultivo utilizado nos distintos tratamentos. O cultivo do camarão marinho Litopenaeus vannamei em bioflocos na densidade de 165 camarões.m–3 pode ser realizado em águas com alcalinidade entre 40 a 160 mg.L–1 de CaCO3, sem comprometer os índices zootécnicos do cultivo.

Parâmetros genéticos para características produtivas e biométricas em abelha Melipona quadrifasciata anthidioides LEPELETIER / Genetic parameters for production and biometrics traits in honey bee Melipona quadrifasciata anthidioides LEPELETIER

Os objetivos deste trabalho foram estimar componentes de variância genética aditiva, fenotípica e residual e a herdabilidade para características relacionadas com a produção de mel e com a estrutura do ninho de abelhas Melipona quadrifasciata anthidioides. Sessenta colônias de diferentes regiões da Bahia foram transladadas para caixas padronizadas modelo INPA e divididas, originando as gerações parentais G1 e G2. Foram medidas as características: estimativa da produção de mel; número, largura, volume e altura dos potes de mel; número, altura e diâmetro dos potes de pólen; peso; número, largura e diâmetro dos discos de cria e estimativa da população da colônia. As medidas foram corrigidas para o efeito fixo de mês de mensuração. Os componentes de variância e herdabilidade foram estimados por meio do método de semelhança entre parentes, utilizando-se abordagem Bayesiana. As médias e os desvios padrão variaram de 2,01±0,70cm para diâmetro de potes de pólen a 2.333,0±384,1kg para o peso das caixas. Houve indicação de convergência para todas as cadeias obtidas. As estimativas de variância genética aditiva variaram de 0,02cm para as características largura dos potes de mel a 38.587,72kg para o peso. Para as estimativas de variâncias fenotípicas, os valores variaram de 0,05 para a altura dos potes de pólen a 95.136,43kg para o peso; e para as variâncias residuais, os valores encontrados variaram de 0,02 para a variável largura dos potes de mel a 56.548,71kg para o peso. As estimativas de herdabilidade variaram de 0,35 a 0,53. Os resultados demonstraram que as características avaliadas possuem variação genética aditiva que garante boa resposta à seleção.

The aim of this study was to estimate components of genetic variance, phenotypic and residual and heritability for traits related to the production of honey and the nest structure of bee Melipona quadrifasciata anthidioides. Sixty colonies from different regions of Bahia were transferred to standard INPA model boxes and divided, creating the parental generations G1 and G2. The following characteristics were measured: estimated production of honey, number, width, height and volume of the honey pots, number, height and diameter of pollen storage pots, weight, number, length and diameter of the brood combs and estimate the population of the colony. The measurements were corrected for the fixed effects of month of measurement. The variance components and heritability were estimated by the method of similarity between relatives using the Bayesian approach. The mean and standard deviations ranged from 2.01±0.70cm diameter pots for pollen to 2333.0±384.1kg to the weight of the boxes. There was indication of convergence for all the chains obtained. Estimates of the additive genetic variance ranged from 0.02cm to the width characteristics of honey pots to 38587.72kg for weight. For the estimates of phenotypic variance the values ranged from 0.05 for the height of the pollen pots to 95136.43kg for weight; and the residual variance the values varied from 0.02 for the variable width of the honey pots to 56548.71kg for weight. The results showed that the characteristics assessed have additive genetic variation that ensures good response to selection.

Análises das embalagens de plantas medicinais comercializadas em farmácias e drogarias do município de Ijuí/RS / Analyses of the packages of medicinal plants sold in pharmacies in the city of Ijuí - RS, Brazil

RESUMO: O objetivo deste estudo foi analisar as embalagens de plantas medicinais comercializadas em farmácias e drogarias do município de Ijuí/RS, a partir das legislações vigentes sobre o tema. A coleta de dados foi realizada na primeira quinzena do mês de dezembro de 2011 em 13 estabelecimentos farmacêuticos localizados na região central do município, através de formulário elaborado a partir da legislação. Foram selecionadas para análise, plantas medicinais acondicionadas em diferentes tipos de embalagens e marcas. Foram avaliadas 44 embalagens de plantas medicinais, das quais 71% estavam irregulares no que se refere a indicação terapêutica e ao modo de preparo, considerando o preconizado pela RDC 10/10. Embora nem todas as drogas vegetais analisadas estivessem notificadas nesta Resolução, considerou-se o mesmo critério para todas as amostras analisadas. Além disso, 16% dos produtos analisados não apresentavam segurança quanto ao acondicionamento. A nomenclatura popular estava presente em todas as amostras, enquanto a nomenclatura botânica em apenas 75%. Conclui-se que todas as embalagens analisadas apresentaram alguma irregularidade em relação ao que estabelece a RDC nº10/2010, tendo em vista que nenhuma apresentou todos os requisitos exigidos por ela. Os resultados observados demonstram que a ausência das informações devidas nas embalagens ou nos folhetos informativos para orientar os consumidores, pode comprometer o uso seguro das plantas e prejudicar a saúde dos usuários.

ABSTRACT: This study aimed to analyze the packages of medicinal plants sold in pharmacies and drugstores in the city of Ijuí, state of Rio Grande do Sul, Brazil, according to the Brazilian legislation about the subject. Data collection was conducted in December 2011 in 13 selected pharmaceutical establishments located downtown, and data were collected using a form prepared from the aforementioned resolution. The medicinal plants included in the analysis were from different types of packages and brands. We evaluated 44 packages of medicinal plants, of which 71% were irregular in relation to shape, therapeutic indication and method of preparation. Moreover, 16% of the total products tested had no package security. The popular nomenclature was present in all samples, while the botanical nomenclature, in 75%. We concluded that all analyzed packages were irregular according to the Brazilian RDC 10/2010 because none of them had all of the requirements. The results observed, such as lack of information on the packages or leaflets to guide consumers, may compromise the safe use of plants and endanger the health of users.

Relationship of inflammatory markers and pain in patients with head and neck cancer prior to anticancer therapy

Pain is a common symptom in patients with cancer, including those with head and neck cancer (HNC). While studies suggest an association between chronic inflammation and pain, levels of inflammatory cytokines, such as C-reactive protein (CRP) and tumor necrosis factor-alpha (TNF-α), have not been correlated with pain in HNC patients who are not currently undergoing anticancer treatment. The purpose of this study was to examine the relationship between these inflammatory markers and perceived pain in HNC patients prior to anticancer therapy. The study group consisted of 127 HNC patients and 9 healthy controls. Pain was assessed using the Brief Pain Inventory (BPI), and serum levels of CRP and TNF-α were determined using the particle-enhanced turbidimetric immunoassay (PETIA) and ELISA techniques, respectively. Patients experiencing pain had significantly higher levels of CRP (P<0.01) and TNF-α (P<0.05) compared with controls and with patients reporting no pain. There were significantly positive associations between pain, CRP level, and tumor stage. This is the first study to report a positive association between perceived pain and CRP in HNC patients at the time of diagnosis. The current findings suggest important associations between pain and inflammatory processes in HNC patients, with potential implications for future treatment strategies.

Uso de indicadores indigestíveis obtidos in situ e in vivo para determinar a digestibilidade de nutrientes em equinos / The use of indigestible markers obtained in situ and in vivo to determine digestibility of nutrients in equine

Objetivou-se estudar a viabilização dos indicadores internos, celulose (CELi) e lignina (LIGi) indigestíveis, obtidos pelas técnicas in situ e in vivo, para predizer a digestibilidade de nutrientes em equinos. Foram utilizados quatro cavalos castrados, por meio de delineamento em blocos ao acaso. Os tratamentos consistiram de metodologias de avaliação de digestibilidade, como a coleta total de fezes (CT) e o uso de indicadores internos, CELi e LIGi, obtidos pelas técnicas in situ (IS) na cavidade ruminal de bovinos e in vivo (IV) nos equinos, por meio da técnica do saco de náilon móvel (TSNM), totalizando cinco tratamentos. A ingestão diária de MS foi de 2,0 por cento do peso vivo de feno de coast-cross. A CELi-IV promoveu a melhor taxa de recuperação do indicador, igualando-se à CT, em 100,31 por cento, enquanto a LIGi resultou nas piores taxas, na média 76,45 por cento, para ambas as técnicas (P<0,05). As estimativas dos coeficientes de digestibilidade dos nutrientes foram adequadamente preditas pela CELi, obtidos IS e IV em que os valores observados foram de 46,41, 48,16, 46,90, 47,92 e 45,51 por cento para MS, MO, PB, FDN e EB, respectivamente. Concluiu-se que a CELi pode ser obtida in vivo por meio da TSNM em equinos, para predizer a digestibilidade de nutrientes...

The objective of this paper was to study the use of the internal indigestible cellulose (CELi) and lignin (LIGi) markers, obtained in situ or in vivo, to predict the apparent digestibility of nutrients in horses. Four castrated horses were utilized in a randomized blocks experimental design. Treatments consisted of different digestibility determination methodologies: total feces collection (TC) and use of CELi and LIGi internal markers obtained by the in situ (IS) incubation in bovine rumen and in vivo (IV) by the mobile nylon bag technique (MNBT) with horses, resulting in five treatments. The diet consisted exclusively of coast-cross hay and the average daily dry matter intake corresponded to 2.0 percent of the BW of the horses. The CELi-IV resulted in the best marker recuperation rate (100.31 percent), being similar to the control method; while the LIGi resulted in low recuperation rates (76.45 percent) for both techniques (P<0.05). The nutrients' digestibility coefficients were effectively predicted by CELi, obtained in situ or in vivo, with average values of 46.41, 48.16, 46.90, 47.92 and 45.51 percent for DM, organic matter, crude protein, neutral detergent fiber and crude energy, respectively. It can be concluded that CELi may be obtained in vivo by MNBT in horses to predict the nutrients' digestibility coefficients...

Therapeutic concentration of morphine reduces oxidative stress in glioma cell line

Morphine is a potent analgesic opioid used extensively for pain treatment. During the last decade, global consumption grew more than 4-fold. However, molecular mechanisms elicited by morphine are not totally understood. Thus, a growing literature indicates that there are additional actions to the analgesic effect. Previous studies about morphine and oxidative stress are controversial and used concentrations outside the range of clinical practice. Therefore, in this study, we hypothesized that a therapeutic concentration of morphine (1 μM) would show a protective effect in a traditional model of oxidative stress. We exposed the C6 glioma cell line to hydrogen peroxide (H2O2) and/or morphine for 24 h and evaluated cell viability, lipid peroxidation, and levels of sulfhydryl groups (an indicator of the redox state of the cell). Morphine did not prevent the decrease in cell viability provoked by H2O2 but partially prevented lipid peroxidation caused by 0.0025% H2O2 (a concentration allowing more than 90% cell viability). Interestingly, this opioid did not alter the increased levels of sulfhydryl groups produced by exposure to 0.0025% H2O2, opening the possibility that alternative molecular mechanisms (a direct scavenging activity or the inhibition of NAPDH oxidase) may explain the protective effect registered in the lipid peroxidation assay. Our results demonstrate, for the first time, that morphine in usual analgesic doses may contribute to minimizing oxidative stress in cells of glial origin. This study supports the importance of employing concentrations similar to those used in clinical practice for a better approximation between experimental models and the clinical setting.