Anti-inflammatory potential of Portuguese thermal waters.

In light of Medical Hydrology, thermal waters (TW) are all-natural mineral waters that emerge inside a thermal resort and have therapeutic applications. Their beneficial effect has been empirically recognized for centuries, being indicated for symptom alleviation and/or treatment of several diseases, almost all associated with inflammation. Indeed, an anti-inflammatory effect has been attributed to many different Portuguese TW but there is no scientific validation supporting this empiric knowledge. In the present study, we aimed to investigate the anti-inflammatory properties of 14 TW pertaining to thermal centers located in the Central Region of Portugal, and grouped according to their ionic profile. Mouse macrophage cells stimulated with lipopolysaccharide (LPS), a Toll-like receptor 4 agonist, were exposed to culture medium prepared in TW. Metabolism, nitric oxide (NO) production, inducible nitric oxide synthase (iNOS) expression levels and the scavenging capacity of TW, were investigated in vitro. 11 out of 14 TW reduced NO production and/or iNOS expression, and/or scavenging activity, in macrophages exposed to LPS. The sulphated/calcic TW did not show any effect on at least one of the inflammatory parameters evaluated. Two sulphurous/bicarbonate/sodic TW and the sulphurous/chlorinated/sodic TW promoted an increase in NO production and/or iNOS expression. Our results validate, for the first time, the anti-inflammatory properties of Portuguese TW, supporting their therapeutic use in the treatment of inflammation-related diseases and promoting their putative application in cosmetic products and medical devices.

Exogenous fibrolytic enzymes and recombinant bacterial expansins synergistically improve hydrolysis and in vitro digestibility of bermudagrass haylage.

Four experiments were conducted to examine the effects of a recombinant bacterial expansin-like protein (BsEXLX1) from Bacillus subtilis and a commercial exogenous fibrolytic enzyme (EFE) preparation for ruminants on hydrolysis of pure substrates (cellulose and xylan) and in vitro digestibility of bermudagrass haylage (BMH). Recombinant Escherichia coli BL21 strain was used to express BsEXLX1; the protein was purified using an affinity column. In experiment 1, carboxymethylcellulose, Whatman #1 filter paper (General Electric, Boston, MA) and oat-spelt xylan substrates were subjected to 4 treatments (1) sodium citrate buffer (control), (2) BsEXLX1 (162 µg/g of substrate), (3) EFE (2.3 mg/g of substrate), and (4) EFE + BsELX1 in 3 independent runs. Samples were incubated at optimal conditions for both additives (pH 5 and 50°C) or at ruminal (pH 6 and 39°C) or ambient (pH 6 and 25°C) conditions for 24 h and sugar release was measured. In experiment 2, digestibility in vitro of BMH was examined after treatment with the following: (1) control (buffer only), (2) BsEXLX1 (162 µg/g of dry matter), (3) EFE (2.2 mg/g of dry matter), and (4) EFE + BsEXLX1 in 3 independent runs at 39°C for 24 h. Experiment 3 examined effects of EFE and BsEXLX1 on simulated preingestive hydrolysis and profile of released sugars from BMH after samples were suspended in deionized water with sodium azide at 25°C for 24 h in 2 independent runs. In experiment 4, the sequence of the BsEXLX1 purified protein was compared with 447 ruminal bacterial genomes to identify similar proteins from the rumen. In experiment 1, compared with EFE alone, EFE and BsEXLX1 synergistically increased sugar release from carboxymethylcellulose and Whatman #1 filter paper under all simulated conditions; however, hydrolysis of xylan was not improved. In experiment 2, compared with EFE alone, treatment with EFE and BsEXLX1 increased neutral detergent fiber and acid detergent fiber digestibility of bermudagrass haylage (by 5.5 and 15%, respectively) and total volatile fatty acid concentrations, and decreased acetate-propionate ratio. In experiment 3, compared with EFE alone. The EFE and BsEXLX1 synergistically reduced concentrations of neutral detergent fiber and acid detergent fiber and increased release of sugars by 9.3%, particularly cellobiose (72.5%). In experiment 4, a similar sequence to that of BsEXLX1 was identified in Bacillus licheniformis, and similar hypothetical protein sequences were identified in Ruminococcus flavefaciens strains along with different protein structures in E. xylanophilum and Lachnospiraceae. This study showed that an expansin-like protein synergistically increased the hydrolysis of pure cellulose substrates and the hydrolysis and digestibility in vitro of BMH.

Suplementação proteica de vacas leiteiras mantidas em pastagem de Tifton 85 durante o período de seca / Protein supplementation of dairy cows grazing Tifton 85 during the dry season

Concentrados com alta proteína bruta e minerais e com menor concentração de energia poderiam ser utilizados em menores quantidades no balanceamento de dietas com volumosos tropicais. Assim, objetivou-se avaliar o consumo de matéria seca, a resposta produtiva e econômica de vacas da raça Girolando mantidas em pastagem de Tifton 85 durante o período de seca, as quais receberam diferentes concentrações de suplementação proteica. Foram utilizadas 12 vacas da raça Girolando em lactação, com produção de leite média inicial de 18,00±1,615kg/dia. Os animais foram distribuídos em três quadrados latinos (4x4), constituídos de quatro tratamentos e quatro períodos experimentais, com duração de 15 dias cada. Os tratamentos consistiram de uma dieta controle, apenas sal mineral e três níveis de suplementação proteica com 40; 50 e 60% de proteína bruta (PB) sendo fornecidas nas quantidades de 2,5kg/animal/dia. Observou-se que os fornecimentos de suplementos proteicos proporcionaram um efeito substitutivo da forragem em relação à suplementação mineral. O uso de suplementação concentrada com teor de 40% de proteína bruta foi mais eficiente em termos produtivos, econômicos e de produção de proteína bruta no leite.(AU)

Concentrates with high crude protein and minerals and lower energy concentration could be used in smaller quantities in balancing diets with tropical bulk. The objective of this study was to evaluate the dry matter intake, the productive and economical response of Girolando cows kept in Tifton 85 pasture during dry season receiving different concentrations of protein supplementation. Twelve lactating Girolando cows with initial mean milk yield of 18.00±1.615kg/day were used. The animals were distributed in three Latin squares (4x4), consisting of four treatments and four experimental periods, each lasting 15 days. The treatments were composed of control diet with only mineral salt, and three levels of protein supplementation with 40, 50, and 60% crude protein (CP) being supplied in the amounts of 2.5kg/animal/day. It was found that the protein supplement supplies provided a substitutive effect of forage relative to mineral supplementation. The use of concentrated supplementation with 40% crude protein content was more efficient in terms of production, production of crude protein in milk and economically.(AU)

Effect of adding clay with or without a Saccharomyces cerevisiae fermentation product on the health and performance of lactating dairy cows challenged with dietary aflatoxin B1.

The study was conducted to examine the effect of supplementing bentonite clay with or without a Saccharomyces cerevisiae fermentation product (SCFP; 19 g of NutriTek + 16 g of MetaShield, both from Diamond V, Cedar Rapids, IA) on the performance and health of dairy cows challenged with aflatoxin B1 (AFB1). Twenty-four lactating Holstein cows (64 ± 11 d in milk) were stratified by parity and milk production and randomly assigned to 1 of 4 treatment sequences. The experiment had a balanced 4 × 4 Latin square design with 6 replicate squares, four 33-d periods, and a 5-d washout interval between periods. Cows were fed a total mixed ration containing 36.1% corn silage, 8.3% alfalfa hay, and 55.6% concentrate (dry matter basis). Treatments were (1) control (no additives), (2) toxin (T; 1,725 µg of AFB1/head per day), (3) T + clay (CL; 200 g/head per day; top-dressed), and (4) CL+SCFP (CL+SCFP; 35 g/head per day; top-dressed). Cows were adapted to diets from d 1 to 25 (predosing period) and then orally dosed with AFB1 from d 26 to 30 (dosing period), and AFB1 was withdrawn from d 31 to 33 (withdrawal period). Milk samples were collected twice daily from d 21 to 33, and plasma was sampled on d 25 and 30 before the morning feeding. Transfer of ingested AFB1 into milk aflatoxin M1 (AFM1) was greater in T than in CL or CL+SCFP (1.65 vs. 1.01 and 0.94%, respectively) from d 26 to 30. The CL and CL+SCFP treatments reduced milk AFM1 concentration compared with T (0.45 and 0.40 vs. 0.75 µg/kg, respectively), and, unlike T, both CL and CL+SCFP lowered AFM1 concentrations below the US Food and Drug Administration action level (0.5 µg/kg). Milk yield tended to be greater during the dosing period in cows fed CL+SCFP compared with T (39.7 vs. 37.7 kg/d). Compared with that for T, plasma glutamic oxaloacetic transaminase concentration, indicative of aflatoxicosis and liver damage, was reduced by CL (85.9 vs. 95.2 U/L) and numerically reduced by CL+SCFP (87.9 vs. 95.2 U/L). Dietary CL and CL+SCFP reduced transfer of dietary AFB1 to milk and milk AFM1 concentration. Only CL prevented the increase in glutamic oxaloacetic transaminase concentration, and only CL+SCFP prevented the decrease in milk yield caused by AFB1 ingestion.

Assistive devices: an effective strategy in non-pharmacological treatment for hand osteoarthritis-randomized clinical trial.

This study aims to evaluate the use of assistive devices as a strategy in non-pharmacological treatment for hand osteoarthritis (HOA). This is a randomized, prospective, parallel, assessor-blinded clinical trial, in which patients with a diagnosis of HOA were randomly allocated to an intervention group (IG), where they received assistive devices for daily life activities, or to a control group (CG), where they received a guideline leaflet with information on joint protection and disease features. The primary outcomes considered were occupational performance, measured by the Canadian Occupational Performance Measure (COPM), and hand function was evaluated through the Score for the Assessment and Quantification of Chronic Rheumatoid Affections of the Hands (SACRAH). The secondary outcomes were pain, measured by the visual analog scale (VAS), and quality of life, measured by the World Health Organization Quality of Life Instrument, Short Form (WHOQOL-BREF). We compared both outcomes before and after interventions and outcomes between groups. Participants from the two groups were assessed at the time of inclusion in the study, 30, and 90 days after initial evaluation. Out of the 39 patients included, 19 were allocated to the IG and 20 to the CG. Only two patients from the CG did not complete the follow-up period. The patients' hand function and occupational performance improved after intervention (30 days-SACRAH-p < 0.05; COPM-p < 0.05; VAS-p < 0.05). When comparing results between the groups, there was a statistical difference in COPM (performance-p < 0.001; and satisfaction-p < 0.001), in the first reevaluation carried out. The use of assistive devices has proved to be an effective alternative in non-pharmacological treatment for HOA. CLINICAL TRIAL REGISTRATION: NCT02667145.

Production, milk and plasma fatty acid profile, and nutrient utilization in Jersey cows fed flaxseed oil and corn grain with different particle size.

We aimed to compare the effects of ground (GC) or cracked corn (CC), with or without flaxseed oil (FSO), on milk yield, milk and plasma fatty acid (FA) profile, and nutrient digestibility in Jersey cows fed diets formulated to contain similar starch concentrations. Twelve multiparous organic-certified Jersey cows averaging (mean ± standard deviation) 455 ± 41.9 kg of body weight and 152 ± 34 d in milk and 4 primiparous organic-certified Jersey cows averaging (mean ± standard deviation) 356 ± 2.41 kg of body weight and 174 ± 30 d in milk in the beginning of the experiment were used. Cows were randomly assigned to treatment sequences in a replicated 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments. Each period lasted 24 d with 18 d for diet adaptation and 6 d for data and sample collection. Treatments were fed as total mixed rations consisting of (dry matter basis): (1) 0% FSO + 27.1% GC, (2) 0% FSO + 28.3% CC, (3) 3% FSO + 27.1% GC, and (4) 3% FSO + 28.3% CC. All cows were offered 55% of the total diet dry matter as mixed grass-legume baleage and treatments averaged 20% starch. Significant FSO × corn grain particle size interactions were observed for some variables including milk concentration of lactose and proportions of cis-9,cis-12,cis-15 18:3 in milk and plasma. The proportion of cis-9,cis-12,cis-15 18:3 in milk and plasma decreased slightly when comparing GC versus CC in 0% FSO cows, but a larger reduction was observed in 3% FSO cows. Dry matter intake did not differ and averaged 16.1 kg/d across diets. Feeding 3% FSO increased yields of milk and milk fat and lactose and feed and milk N efficiencies, but decreased fat, true protein, and MUN concentrations and apparent total-tract digestibility of fiber. The Σ branched-chain, Σ<16C, Σ16C, and Σn-6 FA decreased, whereas Σ18C, Σcis-18:1, and Σtrans-18:1 FA increased in 3% versus 0% FSO cows. No effect of corn particle size was observed for production and milk components. However, the apparent total-tract digestibility of starch was greater in GC than CC cows. Compared with CC, GC increased Σ branched-chain, Σ<16C, Σ16C, Σn-6 FA, and decreased Σ18C and Σ cis-18:1 FA in milk fat. Overall, results of this study are more directly applicable to dairy cows fed low starch, mixed grass-legume baleage-based diets.

Respostas produtivas, marginais e econômicas de vacas Girolando sob pastejo de Tifton 85 recebendo diferentes concentrados / Productive, marginal and economic responses of Girolando cows under grazing of Tifton 85 receiving different concentrates

A energia é considerada o nutriente mais limitante para produção de leite de vacas em pastagens, mas será que a suplementação deve ser energética? Assim, objetivou-se avaliar a resposta produtiva, marginal e econômica de vacas Girolandas mantidas em pastagem de Tifton 85 durante o período das águas, as quais receberam diferentes tipos de suplementação. O primeiro tratamento foi a suplementação mineral ad libitum; o segundo tratamento, o fornecimento de 1,0kg/dia de suplementação proteica; o terceiro e quarto tratamentos, o fornecimento de 2,0kg/dia de suplementação proteico-energética e 4,0kg/dia de suplementação energética, respectivamente. Foram utilizadas 12 vacas com produção de leite média de 15,0±1,99kg/dia. Os dados foram avaliados como três quadrados latinos 4 x 4 simultâneos ao nível de 5% de significância. O consumo de matéria seca de forragem não foi influenciado pelas suplementações (P>0,05). A maior produção de leite (P<0,05) foi observada com a suplementação energética, em média 14,73kg/dia, não diferente da produção com a suplementação proteico-energética, média de 13,84kg de leite/dia. Observaram-se respostas produtivas marginais de 1,03; 0,84 e 0,44kg de leite/kg de concentrado, para suplemento proteico, proteico-energético e energético, respectivamente, bem como saldo financeiro positivo para as suplementações proteica e proteico-energética em relação à suplementação mineral. A suplementação proteico-energética mostrou-se a mais eficiente.(AU)

Energy is considered the most limiting nutrient for the production of milk cows in pastures, but will that supplementation must be energy? The objective was to evaluate the productive, marginal and economic response of Girolando cows kept in Tifton 85 pasture during rain period receiving different types of supplementation. One treatment was mineral supplementation ad libitum, a second treatment providing 1.0kg/day of protein supplementation, and a third and fourth treatments providing 2.0kg/day of protein-energetic supplementation and 4.0kg/day of energy supplementation, respectively. Twelve cows were used with average milk production 15.0±1.99kg/day. The data were evaluated as three Latin squares 4 x 4 simultaneous to the 5% level of significance. The dry matter intake of forage not influenced by supplementation (P> 0.05). The higher milk production was observed with energetic supplementation on average 14.73kg of milk/day, no different of production obtained from protein-energy supplementation, average of 13.84kg of milk/day. Marginal productive response was observed, with responses of 1.03; 0.84 and 0.44kg milk/kg concentrate for proteic supplement, proteic-energetic, and energetic, respectively. Positive financial balance was found for protein and protein-energy supplements in relation to mineral supplementation. Protein-energy supplementation proved the most efficient.(AU)

ADME studies and preliminary safety pharmacology of LDT5, a lead compound for the treatment of benign prostatic hyperplasia.

This study aimed to estimate the absorption, distribution, metabolism and excretion (ADME) properties and safety of LDT5, a lead compound for oral treatment of benign prostatic hyperplasia that has previously been characterized as a multi-target antagonist of α1A-, α1D-adrenoceptors and 5-HT1A receptors. The preclinical characterization of this compound comprised the evaluation of its in vitro properties, including plasma, microsomal and hepatocytes stability, cytochrome P450 metabolism and inhibition, plasma protein binding, and permeability using MDCK-MDR1 cells. De-risking and preliminary safety pharmacology assays were performed through screening of 44 off-target receptors and in vivo tests in mice (rota-rod and single dose toxicity). LDT5 is stable in rat and human plasma, human liver microsomes and hepatocytes, but unstable in rat liver microsomes and hepatocytes (half-life of 11 min). LDT5 is highly permeable across the MDCK-MDR1 monolayer (Papp ∼32×10-6 cm/s), indicating good intestinal absorption and putative brain penetration. LDT5 is not extensively protein-bound and is a substrate of human CYP2D6 and CYP2C19 but not of CYP3A4 (half-life >60 min), and did not significantly influence the activities of any of the human cytochrome P450 isoforms screened. LDT5 was considered safe albeit new studies are necessary to rule out putative central adverse effects through D2, 5-HT1A and 5-HT2B receptors, after chronic use. This work highlights the drug-likeness properties of LDT5 and supports its further preclinical development.

Aqueous extract of Chrysobalanus icaco leaves, in lower doses, prevent fat gain in obese high-fat fed mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Due to the rise in obesity, the necessity for resources and treatments that could reduce the morbidity and mortality associated to this pandemia has emerged. The development of new anti-obesity drugs through herbal sources has been increasing in the past decades which are being used not only as medicine but also as food supplements. Previous studies with the aqueous extract of Chrysobalanus icaco L (AECI) have demonstrated activity on lowering blood glucose levels and body weight. AIM OF THE STUDY: Investigate C. icaco effects in overall adiposity and glycemic homeostasis. MATERIAL AND METHODS: C57BL/6J mice were randomly assigned to standard chow (SC) or high-fat diet (HFD) and treated with AECI in 0.35mg/mL or 0.7mg/mL concentrations ad libitum. Food intake, feed efficiency, metabolic efficiency, body, fat pads and gastrocnemius weight, adiposity index, serum lipids, fecal lipid excretion, locomotor activity in the open field test and insulin and glucose tolerance tests were analyzed and compared. The major components of the extract were demonstrated through HPLC and its antioxidant activity analyzed through DPPH and lipid peroxidation. RESULTS: The AECI in the 0.35mg/mL concentration did not affect food intake or body weight. However, it promoted lower adipose tissue gain, TG levels, and fecal lipid excretion, increased locomotor activity and lean mass weight, and normalized insulin sensitivity and glucose tolerance. Moreover, AECI showed the presence of myricetin 3-O-glucuronide, rutin, quercitrin and myricitrin and demonstrated high-antioxidant activity. CONCLUSIONS: AECI in lower concentrations can prevent fat storage or enhance fat utilization through the increase of locomotor activity. Also, this reinforces its ability to maintain glucose homeostasis through the normalization of insulin sensitivity and glucose tolerance despite the high-fat diet intake. These activities could be associated to the extract's polyphenol content.

ADME studies and preliminary safety pharmacology of LDT5, a lead compound for the treatment of benign prostatic hyperplasia

This study aimed to estimate the absorption, distribution, metabolism and excretion (ADME) properties and safety of LDT5, a lead compound for oral treatment of benign prostatic hyperplasia that has previously been characterized as a multi-target antagonist of α1A-, α1D-adrenoceptors and 5-HT1A receptors. The preclinical characterization of this compound comprised the evaluation of its in vitro properties, including plasma, microsomal and hepatocytes stability, cytochrome P450 metabolism and inhibition, plasma protein binding, and permeability using MDCK-MDR1 cells. De-risking and preliminary safety pharmacology assays were performed through screening of 44 off-target receptors and in vivo tests in mice (rota-rod and single dose toxicity). LDT5 is stable in rat and human plasma, human liver microsomes and hepatocytes, but unstable in rat liver microsomes and hepatocytes (half-life of 11 min). LDT5 is highly permeable across the MDCK-MDR1 monolayer (Papp ∼32×10-6 cm/s), indicating good intestinal absorption and putative brain penetration. LDT5 is not extensively protein-bound and is a substrate of human CYP2D6 and CYP2C19 but not of CYP3A4 (half-life >60 min), and did not significantly influence the activities of any of the human cytochrome P450 isoforms screened. LDT5 was considered safe albeit new studies are necessary to rule out putative central adverse effects through D2, 5-HT1A and 5-HT2B receptors, after chronic use. This work highlights the drug-likeness properties of LDT5 and supports its further preclinical development.

Effect of L-carnitine on exercise performance in patients with mitochondrial myopathy

Exercise intolerance due to impaired oxidative metabolism is a prominent symptom in patients with mitochondrial myopathy (MM), but it is still uncertain whether L-carnitine supplementation is beneficial for patients with MM. The aim of our study was to investigate the effects of L-carnitine on exercise performance in MM. Twelve MM subjects (mean age±SD=35.4±10.8 years) with chronic progressive external ophthalmoplegia (CPEO) were first compared to 10 healthy controls (mean age±SD=29±7.8 years) before they were randomly assigned to receive L-carnitine supplementation (3 g/daily) or placebo in a double-blind crossover design. Clinical status, body composition, respiratory function tests, peripheral muscle strength (isokinetic and isometric torque) and cardiopulmonary exercise tests (incremental to peak exercise and at 70% of maximal), constant work rate (CWR) exercise test, to the limit of tolerance [Tlim]) were assessed after 2 months of L-carnitine/placebo administration. Patients with MM presented with lower mean height, total body weight, fat-free mass, and peripheral muscle strength compared to controls in the pre-test evaluation. After L-carnitine supplementation, the patients with MM significantly improved their Tlim (14±1.9 vs 11±1.4 min) and oxygen consumption ( V ˙ O 2 ) at CWR exercise, both at isotime (1151±115 vs 1049±104 mL/min) and at Tlim (1223±114 vs 1060±108 mL/min). These results indicate that L-carnitine supplementation may improve aerobic capacity and exercise tolerance during high-intensity CWRs in MM patients with CPEO.

Effect of L-carnitine on exercise performance in patients with mitochondrial myopathy.

Exercise intolerance due to impaired oxidative metabolism is a prominent symptom in patients with mitochondrial myopathy (MM), but it is still uncertain whether L-carnitine supplementation is beneficial for patients with MM. The aim of our study was to investigate the effects of L-carnitine on exercise performance in MM. Twelve MM subjects (mean age±SD=35.4±10.8 years) with chronic progressive external ophthalmoplegia (CPEO) were first compared to 10 healthy controls (mean age±SD=29±7.8 years) before they were randomly assigned to receive L-carnitine supplementation (3 g/daily) or placebo in a double-blind crossover design. Clinical status, body composition, respiratory function tests, peripheral muscle strength (isokinetic and isometric torque) and cardiopulmonary exercise tests (incremental to peak exercise and at 70% of maximal), constant work rate (CWR) exercise test, to the limit of tolerance [Tlim]) were assessed after 2 months of L-carnitine/placebo administration. Patients with MM presented with lower mean height, total body weight, fat-free mass, and peripheral muscle strength compared to controls in the pre-test evaluation. After L-carnitine supplementation, the patients with MM significantly improved their Tlim (14±1.9 vs 11±1.4 min) and oxygen consumption ( V ˙ O 2 ) at CWR exercise, both at isotime (1151±115 vs 1049±104 mL/min) and at Tlim (1223±114 vs 1060±108 mL/min). These results indicate that L-carnitine supplementation may improve aerobic capacity and exercise tolerance during high-intensity CWRs in MM patients with CPEO.

Performance of lactating crossbred cows on pasture supplemented with conventional urea or slow release urea / Desempenho de vacas mestiças lactantes em pasto suplementadas com ureia convencional ou ureia de liberação lenta

The effect of using conventional urea (CU) or slow release urea (SRU) was evaluated by replacing soybean meal (SBM) in concentrated supplements in levels of 2, 4 or 6% (dry matter basis) on productive performance of crossbred Holstein x Zebu lactating dairy cows (499±61kg body weight and 167 days of lactation) grazing on elephant grass (11.5% CP and 60% NDF) under rotational grazing during the rainy season. A supplement control (no urea) was used containing SBM as a protein source. A total of 21 cows were distributed an incomplete randomized block design with three periods of 21 days each (14 days of adaptation and seven days of collection). The animals entered the paddocks with a pasture height of 110-120cm and left when the grass reached the height of 40-50cm. The concentrated isonitrogenous supplements (24% crude protein, dry matter basis) were provided in the amount of 3.2kg/cow/day (fed basis). There was no effect (P>0.05) on source of crude protein (SBM vs source NPN), source NPN, level of NPN, interaction between source NPN and level of NPN on milk production (10.0kg/day), fat milk production corrected to 3.5% (10.7kg/day), levels of fat (4.01%), protein (3.66%), lactose (4.16%), total solids (12.86%) and non-fat solids (8.60%) in milk. The replacement of CU by SRU does not promote improvement in the productive performance of crossbred dairy cows grazing on elephant grass during the rainy season. Urea (CU or SRU) can be included in up to 6% of the DM concentrated supplements, replacing SBM, without affecting the productive performance of crossbred cows (Holstein x Zebu) in pasture during the rainy season...

Avaliou-se o efeito da utilização de ureia convencional (UC) ou de ureia de liberação lenta (ULL) em suplementos concentrados, nos níveis de 2, 4 ou 6% (base da matéria seca), em substituição ao farelo de soja, sobre o desempenho produtivo de vacas mestiças Holandês x Zebu (499±61kg de peso corporal e 167 dias de lactação) mantidas em pastos de capim-elefante (11,5% de proteína bruta e 60% de FDNcp), sob lotação intermitente, no período das chuvas. Um suplemento controle (sem ureia) foi utilizado contendo farelo de soja como fonte proteica. Foram utilizadas 21 vacas, distribuídas em delineamento em blocos incompletos balanceados, com três períodos de 21 dias cada (14 dias de adaptação e sete dias de coleta). Os animais entraram nos piquetes com altura do pasto de 110-120cm e saíram quando atingiram altura de 40-50cm. Os suplementos concentrados isonitrogenados (24% de proteína bruta, base da matéria seca) foram fornecidos na quantidade de 3,2kg/vaca/dia (base da matéria natural). Não houve efeito (P>0,05) de fonte de proteína bruta (farelo de soja vs. fonte de NNP), de interação entre fonte de NNP (UC vs. ULL) e nível de NNP (2, 4 e 6% na MS do suplemento), de fonte de NNP e de nível de NNP e sobre a produção de leite (10,0kg/dia), produção de leite corrigida para 3,5% de gordura (10,7kg/dia), teores de gordura (4,01%), proteína (3,66%), lactose (4,16%), extrato seco total (12,86%) e extrato seco desengordurado (8,60%) no leite. A substituição da ureia convencional pela ULL não promove melhoria no desempenho produtivo de vacas leiteiras mestiças em pastagem de capim-elefante, no período das chuvas. A ureia (convencional ou de liberação lenta) pode ser incluída em até 6% na MS de suplementos concentrados, em substituição ao farelo de soja, sem afetar o desempenho produtivo de vacas mestiças (Holandês x Zebu) em pastagem, no período das chuvas...

Two Kunitz-type inhibitors with activity against trypsin and papain from Pithecellobium dumosum seeds: purification, characterization, and activity towards pest insect digestive enzyme.

Two trypsin inhibitors (called PdKI-3.1 and PdKI-3.2) were purified from the seeds of the Pithecellobium dumosum tree. Inhibitors were obtained by TCA precipitation, affinity chromatography on Trypsin-Sepharose and reversed-phase-HPLC. SDS-PAGE analysis with or without reducing agent showed that they are a single polypeptide chain, and MALDI-TOF analysis determined molecular masses of 19696.96 and 19696.36 Da, respectively. The N-terminal sequence of both inhibitors showed strong identity to the Kunitz family trypsin inhibitors. They were stable over a wide pH (2-9) and temperature (37 to 100 degrees C) range. These inhibitors reduced over 84% of trypsin activity with inhibition constant (Ki) of 4.20 x 10(-8) and 2.88 x 10(-8) M, and also moderately inhibited papain activity, a cysteine proteinase. PdKI-3.1 and PdKI-3.2 mainly inhibited digestive enzymes from Plodia interpunctella, Zabrotes subfasciatus and Ceratitis capitata guts. Results show that both inhibitors are members of the Kunitz-inhibitor family and that they affect the digestive enzyme larvae of diverse orders, indicating a potential insect antifeedant.

Anti-inflammatory and antinociceptive activities of an acid fraction of the seeds of Carpotroche brasiliensis (Raddi) (Flacourtiaceae).

Carpotroche brasiliensis is a native Brazilian tree belonging to the Oncobeae tribe of Flacourtiaceae. The oil extracted from its seeds contains as major constituents the same cyclopentenyl fatty acids hydnocarpic (40.5%), chaulmoogric (14.0%) and gorlic (16.1%) acids found in the better known chaulmoogra oil prepared from the seeds of various species of Hydnocarpus (Flacourtiaceae). These acids are known to be related to the pharmacological activities of these plants and to their use as anti-leprotic agents. Although C. brasiliensis oil has been used in the treatment of leprosy, a disease that elicits inflammatory responses, the anti-inflammatory and analgesic activities of the oil and its constituents have never been characterized. We describe the anti-inflammatory and antinociceptive activities of C. brasiliensis seed oil in acute and chronic models of inflammation and in peripheral and central nociception. The mixture of acids from C. brasiliensis administered orally by gavage showed dose-dependent (10-500 mg/kg) anti-inflammatory activity in carrageenan-induced rat paw edema, inhibiting both the edema by 30-40% and the associated hyperalgesia. The acid fraction (200 mg/kg) also showed significant antinociceptive activity in acetic acid-induced constrictions (57% inhibition) and formalin-induced pain (55% inhibition of the second phase) in Swiss mice. No effects were observed in the hot-plate (100 mg/kg; N = 10), rota-road (200 mg/kg; N = 9) or adjuvant-induced arthritis (50 mg/kg daily for 7 days; N = 5) tests, the latter a chronic model of inflammation. The acid fraction of the seeds of C. brasiliensis which contains cyclopentenyl fatty acids is now shown to have significant oral anti-inflammatory and peripheral antinociceptive effects.

Anti-inflammatory and antinociceptive activities of an acid fraction of the seeds of Carpotroche brasiliensis (Raddi) (Flacourtiaceae)

Carpotroche brasiliensis is a native Brazilian tree belonging to the Oncobeae tribe of Flacourtiaceae. The oil extracted from its seeds contains as major constituents the same cyclopentenyl fatty acids hydnocarpic (40.5 percent), chaulmoogric (14.0 percent) and gorlic (16.1 percent) acids found in the better known chaulmoogra oil prepared from the seeds of various species of Hydnocarpus (Flacourtiaceae). These acids are known to be related to the pharmacological activities of these plants and to their use as anti-leprotic agents. Although C. brasiliensis oil has been used in the treatment of leprosy, a disease that elicits inflammatory responses, the anti-inflammatory and analgesic activities of the oil and its constituents have never been characterized. We describe the anti-inflammatory and antinociceptive activities of C. brasiliensis seed oil in acute and chronic models of inflammation and in peripheral and central nociception. The mixture of acids from C. brasiliensis administered orally by gavage showed dose-dependent (10-500 mg/kg) anti-inflammatory activity in carrageenan-induced rat paw edema, inhibiting both the edema by 30-40 percent and the associated hyperalgesia. The acid fraction (200 mg/kg) also showed significant antinociceptive activity in acetic acid-induced constrictions (57 percent inhibition) and formalin-induced pain (55 percent inhibition of the second phase) in Swiss mice. No effects were observed in the hot-plate (100 mg/kg; N = 10), rota-road (200 mg/kg; N = 9) or adjuvant-induced arthritis (50 mg/kg daily for 7 days; N = 5) tests, the latter a chronic model of inflammation. The acid fraction of the seeds of C. brasiliensis which contains cyclopentenyl fatty acids is now shown to have significant oral anti-inflammatory and peripheral antinociceptive effects.

A serine proteinase inhibitor isolated from Tamarindus indica seeds and its effects on the release of human neutrophil elastase.

Proteinaceous inhibitors with high inhibitory activities against human neutrophil elastase (HNE) were found in seeds of the Tamarind tree (Tamarindus indica). A serine proteinase inhibitor denoted PG50 was purified using ammonium sulphate and acetone precipitation followed by Sephacryl S-300 and Sephadex G-50 gel filtration chromatographies. Inhibitor PG50 showed a Mr of 14.9 K on Sephadex G-50 calibrated column and a Mr of 11.6 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. PG50 had selective activity while cysteine proteinases (papain and bromelain) and serine proteinases (porcine pancreatic elastase and bovine chymotrypsin) were not inhibited, it was strongly effective against serine proteinases such as bovine trypsin and isolated human neutrophil elastase. The IC50 value was determined to be 55.96 microg.mL-1. PG50 showed neither cytotoxic nor haemolytic activity on human blood cells. After pre-incubation of PG50 with cytochalasin B, the exocytosis of elastase was initiated using PAF and fMLP. PG50 exhibited different inhibition on elastase release by PAF, at 44.6% and on release by fMLP, at 28.4%. These results showed that PG50 preferentially affected elastase release by PAF stimuli and this may indicate selective inhibition on PAF receptors.

Spinal muscular atrophy type II (intermediary) and III (Kugelberg-Welander). Evolution of 50 patients with physiotherapy and hydrotherapy in a swimming pool.

We added hydrotherapy to 50 patients with spinal muscular atrophy (SMA) who were being treated with individual conventional physiotherapy. Hydrotherapy performed at an approximate temperature of 30 degrees Celsius, twice a week, for thirty minutes in children and forty-five minutes in adults during a 2-year period. The outcome derived from this combined modality of treatment was rated according to physiotherapeutic evaluations, the MMT (Manual Muscular Test), and the Barthel Ladder. Patients were reevaluated at 2-month intervals. After two years of ongoing treatment, we were able to observe that the deformities in hip, knee and foot were progressive in all SMA Type II patients, and in some Type III. Muscle strength stabilized in most SMA Type III patients, and improved in some. MMT was not done in SMA Type II. In all patients we were able to detect an improvement in the Barthel Ladder scale. This study suggests that a measurable improvement in the quality of daily living may be obtained in patients with SMA Types II and III subjected to conventional physiotherapy when associated with hydrotherapy.

Acute necrotizing myopathy and podophyllin toxicity. Report of a fatal case.

A 21 year old male ingested podophyllin in a suicide attempt. The disorder was marked by seizures, coma, peripheral neuropathy, renal failure and acute necrotizing myopathy, an unusual finding. The coma and systemic disturbances resolved within three weeks. The myopathy resolved in 7 weeks, demonstrating a high capacity of muscle recuperation. The sensorimotor peripheral neuropathy persisted until the patient's death 9 weeks after the ingestion, due to septicemia. This report confirms the transient central neurotoxicity of podophyllin and persistent peripheral neurotoxicity of podophyllin, and describes a reversible necrotizing myopathy associated to mitochondrial abnormalities, a still unreported feature of podophyllin toxicity.