Bariatric surgery induces morphological changes in the extensor digitorum longus muscle in the offspring of obese rats.

INTRODUCTION: The effects of the maternal nutritional environment on the growth and metabolism of the offspring, and its impacts on health in adult life are defined as metabolic programming. Thus, the objective of this study was to evaluate the effects of Roux-en-Y gastric bypass (RYGB) on the morphology of muscle fiber and neuromuscular junction (NMJ) of the offspring of rats submitted to RYGB. METHODS: Three-week-old Wistar rats were separated into two groups: 1) CAF SHAM which received a cafeteria diet and was submitted to a sham operation and 2) CAF RYGB, which received a cafeteria diet and was submitted to RYGB. The first generation (F1) offspring (male) was named according to the treatment of mothers as CAF SHAM-F1 and CAF RYGB-F1 and received a standard diet after weaning. At 17 weeks, the animals were euthanized, and the extensor digitorum longus muscle (EDL) was collected and processed in light microscopy and transmission electron microscopy for morphological and morphometric analysis. RESULTS: The CAF RYGB-F1 group showed a reduction in the weight of the EDL muscle and also a reduction in the area of type I, IIa and IIb fibers and a nucleus/fiber ratio. This same group also showed an increase in the capillary density and myofibrillar disorganization and in the Z-line, as well as a reduction in the area of the NMJs. CONCLUSION: The RYGB surgery in mothers produced morphological changes in the skeletal striated muscles of the offspring.

GH11 xylanase from Aspergillus tamarii Kita: Purification by one-step chromatography and xylooligosaccharides hydrolysis monitored in real-time by mass spectrometry.

The present study describes the one-step purification and biochemical characterization of an endo-1,4-ß-xylanase from Aspergillus tamarii Kita. Extracellular xylanase was purified to homogeneity 7.43-fold through CM-cellulose. Enzyme molecular weight and pI were estimated to be 19.5kDa and 8.5, respectively. The highest activity of the xylanase was obtained at 60°C and it was active over a broad pH range (4.0-9.0), with maximal activity at pH 5.5. The enzyme was thermostable at 50°C, retaining more than 70% of its initial activity for 480min. The K0.5 and Vmax values on beechwood xylan were 8.13mg/mL and 1,330.20µmol/min/mg of protein, respectively. The ions Ba2+ and Ni2+, and the compounds ß-mercaptoethanol and DTT enhanced xylanase activity, while the heavy metals (Co2+, Cu2+, Hg+, Pb2+ and Zn2+) strongly inhibited the enzyme, at 5mM. Enzymatic hydrolysis of xylooligosaccharides monitored in real-time by mass spectrometer showed that the shortest xylooligosaccharide more efficiently hydrolyzed by A. tamarii Kita xylanase corresponded to xylopentaose. In agreement, HPLC analyzes did not detect xylopentaose among the hydrolysis products of xylan. Therefore, this novel GH11 endo-xylanase displays a series of physicochemical properties favorable to its application in the food, feed, pharmaceutical and paper industries.

Detection of pathogenic Escherichia coli and microbiological quality of chilled shrimp sold in street markets.

UNLABELLED: Foodborne illnesses caused by Escherichia coli are one of the most important gastrointestinal diseases and therefore represent a public health risk. The presence of E. coli in water or in products such as shrimp indicates faecal contamination. However, indicator micro-organisms can be used to evaluate the microbiological quality of food sold in markets. This study focused on detecting isolates of E. coli containing the genes stx1A, stx2A, eae, LTI, STa, STb, aggR and pCVD432 in chilled shrimp sold in street markets in the municipality of São Paulo, Brazil, and to assess the microbiological quality of this product. Enteropathogenic and enterotoxigenic E. coli pathotypes were detected on the surface of two chilled shrimp samples. Salmonella spp. was not isolated. In addition, contamination of surface and muscle of the shrimp samples was found to be correlated. The detection of EPEC and ETEC pathotypes in chilled shrimp sold in street markets in Brazil provides useful epidemiological information for public health authorities to improve food safety and public health. SIGNIFICANCE AND IMPACT OF THE STUDY: Shrimps are crustaceans commonly produced and consumed in Brazil. Specimens of Farfantepenaeus brasiliensis and Litopenaeus schmitti sold in street markets were examined by PCR to detect the presence of Escherichia coli pathotypes (enteropathogenic, enterotoxigenic, enterohemorrhagic and enteroinvasive). EPEC and ETEC strains were detected in whole shrimp. These findings provide useful information for public health authorities to improve the food safety and health of the Brazilian population.

Shiga toxigenic and enteropathogenic Escherichia coli in water and fish from pay-to-fish ponds.

UNLABELLED: Escherichia coli is part of the normal microflora of the intestines of mammals. However, among the enteric pathogens, it is one of the leading causes of intestinal diseases, especially Shiga toxigenic E. coli, which can cause diarrhoea, haemorrhagic colitis and complications like haemolytic uraemic syndrome and thrombotic thrombocytopaenic purpura. Escherichia coli is considered a serious public health problem. Water and fish samples were subjected to biochemical tests to confirm the presence of E. coli and by PCR to verify the presence of pathogenic strains (O157, enteropathogenic and shiga toxigenic) in water and fish (skin, gastrointestinal tract and muscles) from pay-to-fish ponds located in the Córrego Rico watershed in the northeastern region of the state of São Paulo, Brazil. Of the 115 E. coli isolates from fish or water, five (4·34%) contained eae and stx2 genes, one had only the eae gene and two had the stx1 gene. An isolate containing the stx2 gene was also found in the water sample. In addition, eight isolates (6·95%) from the fish gastrointestinal tract contained rfbEO157:H7 (O157 gene), and three (2·61%) contained stx2 and eae genes, demonstrating the potential risk to the environment and public health. The results provide useful basic information for the proper management of these environments and animals in order to prevent faecal pollution, reducing health risks to the Brazilian population. SIGNIFICANCE AND IMPACT OF THE STUDY: Pay-to-fish ponds are a common commercial activity in Brazil. Samples of water and Oreochromis niloticus were examined by PCR to detect the presence of pathogenic strains of Escherichia coli (O157, enteropathogenic and shiga toxigenic). Several pathogenic strains were detected in this study, providing useful epidemiological information for the proper management of these environments and animals in order to prevent faecal pollution, reducing health risks to the Brazilian population.

Profile of bioactive compounds from grape pomace (Vitis vinifera and Vitis labrusca) by spectrophotometric, chromatographic and spectral analyses.

The aim of this study was to characterize grape pomace (GP) from winemaking byproducts of different grape samples (Cabernet Sauvignon-CS; Merlot-ME; Mix composed of 65% Bordeaux, 25% Isabel and 10% BRS Violet-MI and Terci-TE) with a view to exploiting its potential as a source of bioactive compounds and an alternative to the reuse of waste. Bioactive compounds such as individual phenolic compounds and polyunsaturated fatty acids (PUFA) were identified and quantified by spectrophotometric, chromatographic and spectral analyses. The sample of MI had the highest concentrations for total phenolic compounds and total flavonoids, while TE had the highest content for total monomeric anthocyanins. For all samples it was possible to identify 13 different anthocyanins by high performance liquid chromatography (HPLC) and mass spectrometry (MS). Moreover, the GP samples showed phenolic acids; flavan-3-ols such as catechin; flavonols such as quercetin, rutin and kaempferol; and stilbenes such as trans-resveratrol. Therefore, grape pomace can be considered a source for the recovery of phenolic compounds having antioxidant activity as well as a rich source of PUFA. Thus it can be used as an ingredient in the development of new food products, since it is suitable for human consumption, and a viable alternative both to adding nutritional value to food and to reduce environmental contamination.

Effects of immobilization and remobilization on the ankle joint in Wistar rats

A sprained ankle is a common musculoskeletal sports injury and it is often treated by immobilization of the joint. Despite the beneficial effects of this therapeutic measure, the high prevalence of residual symptoms affects the quality of life, and remobilization of the joint can reverse this situation. The aim of this study was to analyze the effects of immobilization and remobilization on the ankle joint of Wistar rats. Eighteen male rats had their right hindlimb immobilized for 15 days, and were divided into the following groups: G1, immobilized; G2, remobilized freely for 14 days; and G3, remobilized by swimming and jumping in water for 14 days, performed on alternate days, with progression of time and a series of exercises. The contralateral limb was the control. After the experimental period, the ankle joints were processed for microscopic analysis. Histomorphometry did not show any significant differences between the control and immobilized/remobilized groups and members, in terms of number of chondrocytes and thickness of the articular cartilage of the tibia and talus. Morphological analysis of animals from G1 showed significant degenerative lesions in the talus, such as exposure of the subchondral bone, flocculation, and cracks between the anterior and mid-regions of the articular cartilage and the synovial membrane. Remobilization by therapeutic exercise in water led to recovery in the articular cartilage and synovial membrane of the ankle joint when compared with free remobilization, and it was shown to be an effective therapeutic measure in the recovery of the ankle joint.

Cells of the rectum of Bombyx mori affected by experimental inoculation with Alphabaculovirus.

Bombyx mori is an insect whose cocoon is used in the sericulture industry, which is an important activity in parts of southern Brazil. When parasitized by Bombyx mori nucleopolyhedrovirus (BmNPV) of the genus Alphabaculovirus (alphaBV), it develops nuclear polyhedrosis disease. In Brazil, an alphaBV was isolated from larvae of B. mori and various target tissues were identified. However, how this geographic viral isolate affects the rectum of silk moths was unknown. The rectum, a component of the cryptonephric system, acts to absorb water and mineral salts, and its importance for the metabolic balance of insects provoked interest in analyzing how it is affected by BmMNPV infection. Fifth instar B. mori larvae were inoculated with a viral suspension, and from the second to the ninth day post-inoculation, segments of the rectum (anterior and anal canal) were examined using light microscopy. The cryptonephric epithelial cells in the anterior region revealed no evidence of infection. However, from the fifth day post-inoculation, cells of the anal canal showed cytopathologies characteristic of alphaBV. Infection of the anal canal and other surrounding tissues led to tissue disorganization, with accumulation of polyhedra in the perinephric space and compartmentalization of the cryptonephric system, promoting changes in the fecal pellets, signalling physiological changes. These observations contribute to our understanding of the infectious cycle of BmMNPV in B. mori.

Effects of immobilization and remobilization on the ankle joint in Wistar rats.

A sprained ankle is a common musculoskeletal sports injury and it is often treated by immobilization of the joint. Despite the beneficial effects of this therapeutic measure, the high prevalence of residual symptoms affects the quality of life, and remobilization of the joint can reverse this situation. The aim of this study was to analyze the effects of immobilization and remobilization on the ankle joint of Wistar rats. Eighteen male rats had their right hindlimb immobilized for 15 days, and were divided into the following groups: G1, immobilized; G2, remobilized freely for 14 days; and G3, remobilized by swimming and jumping in water for 14 days, performed on alternate days, with progression of time and a series of exercises. The contralateral limb was the control. After the experimental period, the ankle joints were processed for microscopic analysis. Histomorphometry did not show any significant differences between the control and immobilized/remobilized groups and members, in terms of number of chondrocytes and thickness of the articular cartilage of the tibia and talus. Morphological analysis of animals from G1 showed significant degenerative lesions in the talus, such as exposure of the subchondral bone, flocculation, and cracks between the anterior and mid-regions of the articular cartilage and the synovial membrane. Remobilization by therapeutic exercise in water led to recovery in the articular cartilage and synovial membrane of the ankle joint when compared with free remobilization, and it was shown to be an effective therapeutic measure in the recovery of the ankle joint.

Bombyx mori pylorus infection by Alphabaculovirus.

Alphabaculovirus is an entomopathogenic virus genus that infects Bombyx mori, which is known as the Bombyx mori multiple nucleopolyedrovirus (BmMNPV). This virus is polyorganotrophic, and a series of tissues are known as targets; however, there is currently no information regarding infection in the pylorus, the segment of the hindgut that is present in the midgut transition and is responsible for food passage control. Thus, in the present study, we aimed to analyze infection of the B. mori pylorus by BmMNPV. To do so, hybrid B. mori larvae were inoculated with a viral suspension of BmMNPV, and segments of the intestine containing the pylorus and its subdivisions, the posterior interstitial ring (PIR), pyloric cone, and pyloric valve, were dissected and processed for light microscopy on different days post inoculation. The results showed that B. mori pylorus subdivisions respond differently to infection, and the anterior area of the PIR is susceptible with these cells being the secondary infection targets. Cytological analysis revealed the presence of viroplasm in the hypertrophic nucleus, followed by the formation and development of viral polyhedra. Cytolysis occurred at the end of the infectious cycle, thereby releasing polyhedra and enabling the spread of the disease. There was no evidence of BmMNPV infection in the posterior area of the PIR, cone, or pyloric valve. These results will contribute to greater understanding of the virus infectious cycle, whose consequent epizootic disease can negatively impact this economically important insect that is used in silk production in Brazil.

Morphology of the Pylorus of Bombyx mori (Linnaeus) (Lepidoptera: Bombycidae).

The pylorus is a segment of the hindgut of great morphological diversity, which is subdivided into the posterior interstitial ring, pyloric cone, and pyloric valve. There is no morphological information on Bombyx mori (L.) hybrids, which is an insect of economic interest to Brazil. Consequently, this study aimed to analyze the morphology of the pylorus of hybrid larvae of fifth instars of B. mori from crossbreeding of Chinese and Japanese races. These were anesthetized, and the pylorus was fixed and processed by light microscopy and scanning electron microscopy. The pylorus presented as a separate compartment, consisting of a simple epithelium, with large variation in cell morphology, covered on the luminal side by an intima with specializations in some areas, the spicules. The musculature was formed by bundles of striated muscle fibers in longitudinal and circular arrangement. These fibers were multinucleated with central and peripheral nuclei, operating in peristalsis. The morphology of the pylorus is a reflection of functional dynamism, and this study provided a better understanding of its physiology, which promotes the regulated movement of the bolus, preventing reflux and directing it towards the ileum in the formation of fecal pellets.

A designed bifunctional laccase/ß-1,3-1,4-glucanase enzyme shows synergistic sugar release from milled sugarcane bagasse.

A bifunctional enzyme has been created by fusing two Bacillus subtilis enzymes: the ß-1,3-1,4-glucanase (BglS, EC 3.2.1.73) that hydrolyzes plant cell wall ß-glucans and the copper-dependent oxidase laccase (CotA, EC 1.10.3.2) that catalyzes the oxidation of aromatic compounds with simultaneous reduction of oxygen to water. The chimeric laccase/ß-1,3-1,4-glucanase was created by insertion fusion of the bglS and cotA genes, and expressed in Escherichia coli. The affinity-purified recombinant chimeric enzyme showed both laccase and glucanase activities, with a maximum laccase activity at pH 4.5 and 75°C that showed a V(max) 30% higher than observed for the parental laccase. The maximum glucanase activity in the chimeric enzyme was at pH 6.0 and 50°C, with a slight reduction in V(max) by ∼10% compared with the parental glucanase. A decreased K(M) resulted in an overall increase in the K(cat)/K(M) value for the glucanase activity of the chimeric enzyme. The hydrolytic activity of the chimera was 20% higher against natural milled sugarcane bagasse as compared with equimolar mixtures of the separate parental enzymes. Molecular dynamics simulations indicated the approximation of the two catalytic domains in the chimeric enzyme, and the formation of an inter-domain interface may underlie the improved catalytic function.

Virucidal evaluation of sodium dichloroisocyanurate formulation against nucleopolyhedroviruses of silkworm / Avaliação antiviral do formulado de sódio dicloroisocianurato contra o nucleopoliedrovírus do bicho-da-seda

This study evaluated the effectiveness of a disinfectant with low corrosive action and which is not toxic to the environment, the sodium dichloroisocyanurate formulation, on the Bombyx mori nucleopolyhedrovirus (BmNPV). For this, 5th-instar B. mori silkworm larvae were divided into four experimental groups of 4 replicates with 15 larvae each, totalling 60 larvae per group. The groups were fed with mulberry (Morus sp.) leaf discs containing: BmNPV treated with the disinfectant, untreated BmNPV, only the disinfectant, and water (control). The results showed that the disinfectant does not inactivate the BmNPV and also exerts a negative effect on the insect"s resistance.

O estudo avaliou a eficiência de um desinfetante que apresenta baixa ação corrosiva e que não é tóxico ao meio ambiente, o formulado de sódio dicloroisocianurato, sobre o nucleopoliedrovírus Bombyx mori (BmNPV). Para tanto, lagartas do bicho-da-seda, B. mori, de 5º instar foram divididas em quatro grupos experimentais, 4 repetições com 15 lagartas cada, totalizando 60 lagartas por grupo. Os grupos foram alimentados com discos foliares de amoreira (Morus sp.) contendo: o BmNPV tratado com o desinfetante (solução 1); o BmNPV não tratado (solução 2); apenas o desinfetante (solução 3); e água (solução 4, controle). Os resultados mostraram que o desinfetante não inativa o BmNPV e também exerce efeito negativo na resistência do inseto.

Evaluation of rats´ soleus muscle submitted to remobilization protocol with therapeutic ultrasound associated with static stretching

Despite the deleterious effects, immobilization, is still often used in the treatment of musculoskeletal disorders, even with the occurrence of muscular atrophy, its reversal is a major challenge to rehabilitation. The aim of this study was evaluate the tropism of rats soleus muscle submitted to remobilization with static stretching, preceded by ultrasound, thermal and non thermal. We used 28 rats divided into four groups: G1‑immobilized and remobilized with static stretching, G2-remobilized with stretching preceded by ultrasound 1.0 W.cm–²; G3 – ultrasound at 0.5 W.cm–² , G4 – ultrasound with 0.2 W.cm–². All animals were immobilized in plantarflexion, producing shortening of the right soleus muscle for 15 days. For the groups subjected to remobilization with ultrasound doses were used according to the group, for 3 minutes, for 10 days with an interval of 2 days after the 5th treatment. After treatment with ultrasound (or not for the G1), the animals were subjected to 3 sets of 30 seconds, with 30 seconds interval between them, of static stretching of the soleus. The soleus were dissected, weighed and processed for preparation of histological slides in cross section, and evaluated the smallest diameter of 100 fibers per muscle. There was significant reduction in weight between left and right muscles in all groups, for diameters G3 showed no difference. Conclusion: stretching with or without ultrasound, was unable to reverse the deleterious effects of immobilization on muscle weight, but in the mean dose there was protective effect on the diameter of the fibers.

A semi-tethered test for power assessment in running.

We analyzed the usefulness of a semi-tethered field running test (STR) and the relationships between indices of anaerobic power, anaerobic capacity and running performance in 9 trained male sprinters (22.2 ± 2.9 yrs, 176 ± 1 cm, 68.0 ± 9.4 kg). STR involved an all out 120 m run attached to an apparatus that enabled power calculation from force and velocity measures. Subjects also carried out a cycloergometer Wingate Anaerobic Test (WT), an all out 300 m run and had accessed their maximal accumulated oxygen deficit (MAOD) on a treadmill. Peak and mean powers attained in STR (1720 ± 221 and 1391 ± 201 W) were greater but significantly related (r = 0.82; P < 0.01) to those in the WT (808 ± 130 and 603 ± 87 W). In addition, power measures derived from the STR were stronger related to running performance compared to those from the WT (r = 0.81-0.94 vs. 0.68-0.84; P < 0.05). Relationships between MAOD and most power indices were only weak to moderate. These results support the usefulness of STR for specific power assessment in field running and suggest that anaerobic power and capacity are not related entities, irrespective of having been evaluated using similar or dissimilar exercise modes.

Silkworm salivary glands are not susceptible to Bombyx mori nuclear polyhedrosis virus.

A nuclear polyhedrosis virus isolated from infected Bombyx mori, BmNPV, was used to inoculate silkworms to determine salivary gland cell susceptibility. The salivary gland was removed from infected silkworms at different times post-inoculation and examined by light microscopy. The salivary gland cells did not exhibit any signs of BmNPV infection; however, fat body and tracheal cells, used as positive controls, showed characteristic cytopathological changes caused by BmNPV infection, which confirmed inoculum viability. The morphological distribution of tracheal branches and the basal lamina, which serves as a barrier to viral penetration, are apparently involved in this resistance to infection.

Molecular markers in commercial Bombyx mori (Lepidoptera: Bombycidae) hybrids susceptible to multiple nucleopolyhedrovirus.

The silkworm Bombyx mori L. is particularly susceptible to virus diseases, especially B. mori nucleopolyhedrovirus (BmNPV). Disease resistance, along with high productivity, are important selection criteria for developing commercial hybrids of B. mori. We used bioassays and molecular markers linked to susceptibility/resistance to baculovirus infection to analyze the response of commercial B. mori hybrids from two companies to a geographic isolate of B. mori multiple nucleopolyhedrovirus (BmMNPV) from Paraná state in Brazil. Both of these commercial lines were highly susceptible to BmMNPV, with death rates of 92 and 94%. A polymorphic fragment of approximately ~350 bp, associated with susceptibility, and an ~800-bp fragment, associated with resistance to BmMNPV, were detected in all specimens. An additional fragment of ~480 bp, recently identified by our research team as a BmMNPV genomic sequence, was detected in the infected silkworms and could be used as a molecular marker for the diagnosis of nucleopolyhedrovirus infection.

Lactate and glucose minimum speeds and running performance.

This study aimed to analyse the validity of glucose minimum speed (GMS) for lactate minimum speed (LMS) assessment during running and their relationship to endurance performance. Eight male trained runners (28.7 +/- 9.0 years) volunteered to take part in this study and underwent an official 10-km road race and a track lactate minimum test (LMT) (0.5-km sprint plus 6 x 800 m from 87 to 98% of maximal 3-km speed). Lactate and glucose minimum speeds were considered those related to the minimum blood lactate and glucose concentrations respectively attained during the graded phase of LMT. Significant correlations (p < 0.05) were found between LMS and GMS (r = 0.72) and LMS and 10-km performance (r = 0.83), but not between GMS and 10-km performance (r = 0.49). No significant differences (p > 0.05) were found between LMS (4.75 +/- 0.08 m/s), GMS (4.73 +/- 0.07 m/s) and 10-km mean speed (4.79 +/- 0.17 m/s). In conclusion, we found GMS to be a good predictor of LMS during track LMT, LMS being well related to endurance running performance.