Tuberculose e o estudo molecular da sua epidemiologia

Systems that can distinguish epidemiologically-related Mycobacterium tuberculosis strains from unrelated ones are extremely valuable. Molecular biology techniques have allowed a great deal of information to be acquired about the infectious disease tuberculosis (TB) that was very hard or impossible to obtain by conventional epidemiology. A typing method based on bacterial DNA genome differences, known as RFLP (restriction fragment length polymorphism), is widely used to discriminate strains in the epidemiologic study of TB. However, RFLP is laborious and there is a tendency to replace it by other methods. Thus, other DNA sequences have been employed as epidemiological markers, as in Spoligotyping, a fast technique based on PCR followed by differential hybridization of amplified products. The polymorphism observed among different isolates is probably the product of strain-dependent recombination. MIRU (mycobacterial interspersed repetitive unit) typing is a reproducible and fast assay, involving the generation of genotypes based on the study of 12 loci containing VNTRs (variable-number tandem repeats) in strains of the M. tuberculosis complex. It compares strains from different geographic areas and allows the movement of individual lineages to be tracked, as in RFLP. This approach enables a greater number of isolates to be analyzed, leading to the identification of a larger number of foci of transmission within the population and thus to improved ways of slowing the progress of the disease.

Antimycobacterial activity of some Brazilian indigenous medicinal drinks

Tuberculosis (TB) is a very serious problem worldwide and the increasing number of multiple drugs resistant TB cases makes the search for new anti-TB drugs an urgent need. Indigenous knowledge about the use of native plants to treat illnesses has contributed to the discovery of new medicines. In this study, the antimycobacterial activity of seven medicinal drinks was assessed: Ananas sativus (hydroalcoholic fruit extract), Aristolochia triangularis (aqueous and hydroalcoholic leaf, root and stem extracts), Bromelia antiacantha (hydroalcoholic fruit extract), Stryphnodendron adstringens (hydroalcoholic bark extract), Tabebuia ovellanedae (hydroalcoholic bark extract), Vernonia polyanthes (hydroalcoholic root extract), all used by the Vanuíre indigenous community in the treatment of respiratory diseases. The activity was evaluated by using a time-to-kill assay, in which Mycobacterium tuberculosis H37Rv was cultured on Löwenstein-Jensen medium, after thirty minutes, one, three, six, twelve and twenty-four hours contact of the bacteria with each drink. Within half to one hour contact, the hydroalcoholic drinks of A. triangularis, S. adstringens, T. ovellanedae and V. polyanthes reduced the bacterial growth by 2 orders of magnitude in CFU/mL, and all bacterial growth was absent after three hours contact. In contrast, no mycobactericidal effect was detected in the aqueous extract of A. triangularis or in the hydroalcoholic beverages of A. sativus and B. antiacantha, even after twenty-four hours contact.

Modulation of the pharmacological effects of enzymatically-active PLA 2 by BTL-2, an isolectin isolated from the Bryothamnion triquetrum red alga

Background. An interaction between lectins from marine algae and PLA 2 from rattlesnake was suggested some years ago. We, herein, studied the effects elicited by a small isolectin (BTL-2), isolated from Bryothamnion triquetrum, on the pharmacological and biological activities of a PLA 2 isolated from rattlesnake venom (Crotalus durissus cascavella), to better understand the enzymatic and pharmacological mechanisms of the PLA 2 and its complex. Results. This PLA2 consisted of 122 amino acids (approximate molecular mass of 14 kDa), its pI was estimated to be 8.3, and its amino acid sequence shared a high degree of similarity with that of other neurotoxic and enzymatically-active PLA2s. BTL-2 had a molecular mass estimated in approximately 9 kDa and was characterized as a basic protein. In addition, BTL-2 did not exhibit any enzymatic activity. The PLA2 and BTL-2 formed a stable heterodimer with a molecular mass of approximately 24-26 kDa, estimated by molecular exclusion HPLC. In the presence of BTL-2, we observed a significant increase in PLA2 activity, 23% higher than that of PLA2 alone. BTL-2 demonstrated an inhibition of 98% in the growth of the Gram-positive bacterial strain, Clavibacter michiganensis michiganensis (Cmm), but only 9.8% inhibition of the Gram-negative bacterial strain, Xanthomonas axonopodis pv passiflorae (Xap). PLA2 decreased bacterial growth by 27.3% and 98.5% for Xap and Cmm, respectively, while incubating these two proteins with PLA2-BTL-2 inhibited their growths by 36.2% for Xap and 98.5% for Cmm. PLA2 significantly induced platelet aggregation in washed platelets, whereas BTL-2 did not induce significant platelet aggregation in any assay. However, BTL-2 significantly inhibited platelet aggregation induced by PLA2. In addition, PLA 2 exhibited strong oedematogenic activity, which was decreased in the presence of BTL-2. BTL-2 alone did not induce oedema and did not decrease or abolish the oedema induced by the 48/80 compound. Conclusion. The unexpected results observed for the PLA2-BTL-2 complex strongly suggest that the pharmacological activity of this PLA2 is not solely dependent on the presence of enzymatic activity, and that other pharmacological regions may also be involved. In addition, we describe for the first time an interaction between two different molecules, which form a stable complex with significant changes in their original biological action. This opens new possibilities for understanding the function and action of crude venom, an extremely complex mixture of different molecules. © 2008 Oliveira et al; licensee BioMed Central Ltd.

Clinical outcome in chronic myeloid leukemia after allogeneic hematopoietic stem cell transplantation: The experience of the Bone Marrow Transplantation Unit of FUNFARME/BRAZIL using FISH

Investigation of the efficacy of allogeneic hematopoietic stem cell transplantation (allo-HSCT) in chronic myeloid leukemia patients is essential to predict prognosis and survival. In 20 patients treated at the Bone Marrow Transplantation Unit of São José do Rio Preto (São Paulo, Brazil), we used fluorescence in situ hybridization (FISH) to investigate the frequency of cells with BCR/ABL rearrangement at diagnosis and at distinct intervals after allo-HSCT until complete cytogenetic remission (CCR). We investigated the disease-free survival, overall survival in 3 years and transplant-related mortality rates, too. Bone marrow samples were collected at 1, 2, 3, 4, 6, 12, and 24 months after transplantation and additional intervals as necessary. Success rate of the FISH analyses was 100%. CCR was achieved in 75% of the patients, within on average of 3.9 months; 45% patients showed CCR within 60 days after HSCT. After 3 years of the allo-HSCT, overall survival rate was 60%, disease-free survival was 50% and the transplant-related mortality rate was 40%. The study demonstrated that the BCR-ABL FISH assay is useful for follow-up of chronic myeloid leukemia patients after HSCT and that the clinical outcome parameters in our patient cohort were similar to those described for other bone marrow transplantation units. ©FUNPEC-RP.

Subtelomeric region of chromosome 2 in patients with autism spectrum disorders

Autism spectrum disorders are severe psychiatric diseases commonly identified in the population. They are diagnosed during childhood and the etiology has been much debated due to their variations and complexity. Onset is early and characterized as communication and social interaction disorders and as repetitive and stereotyped behavior. Austistic disorders may occur together with various genetic and chromosomal diseases. Several chromosomal regions and genes are implicated in the predisposition for these diseases, in particular those with products expressed in the central nervous system. There are reports of autistic and mentally handicapped patients with submicroscopic subtelomeric alterations at the distal end of the long arm of chromosome 2. Additionally, there is evidence that alterations at 2q37 cause brain malformations that result in the autistic phenotype. These alterations are very small and not identified by routine cytogenetics to which patients are normally submitted, which may result in an underestimation of the diagnosis. This study aimed at evaluating the 2q37 region in patients with autistic disorders. Twenty patients were studied utilizing the fluorescence in situ hybridization technique with a specific probe for 2q37. All of them were also studied by the GTC banding technique to identify possible chromosomal diseases. No alterations were observed in the 2q37 region of the individuals studied, and no patient presented chromosomal diseases. This result may be due to the small sample size analyzed. The introduction of routine analysis of the 2q37 region for patients with autistic disorders depends on further studies. ©FUNPEC-RP.

Influence of the environment on behavior patterns of laying hens kept in cages

The aim of this paper is to verify the correlation between environmental indicators and behaviors expressed by laying hens kept in cages. The birds react to a severe environment through their behaviors, end the behaviors can be monitored to identify the birds' welfare conditions. The behaviors birds display ere the result of stress caused by the combination of environmental temperature, relative humidity, radiant heat, and air speed (environmental temperature being the most important). In order to check the influence of the environment, an experiment was carried out on a commercial poultry farm, located in the city of Bastos. The study was initiated in March 2007, during four non-consecutive weeks. The birds' behaviors were recorded using video, by cameras installed in the cages. The birds behaviors were identified and noted for the frequency of occurrence for each bird, and the average duration of each behavior (in seconds), using video samples of 15 minutes recorded from 1 PM to 4 PM. The environmental variables collected were: air temperature, concentration of ammonia, relative air humidity, velocity of the air, noise, roof temperature, and light intensity. The observed behaviors were: opening wings, stretching, threatening, ruffling feathers, drinking water, aggressive pecking, eating, running, lying down, stretching head out of the cage, preening, mounting and prostrating. Principal Components Analysis was used to determine associations between the behavior variables and environmental variables described above. In this experiment, there were no significant correlations between behavioral variables and environmental variables.

Effects of the density, aviary type, breed, and age on the behaviors of laying hens kept in cages

An experiment was carried out in order to investigate the behaviors of laying hens due to the environmental factors of: density inside of the cage, aviary type, breed, and age. The experiment was configured as a factorial 4x2x2x2 study, with treatments being four different ages, two different breeds, two different cage densities, and two different aviaries. The birds' behaviors were recorded using video cameras installed in the cages, using samples of 15 minutes recorded from 12 PM to 4 PM. The observed behaviors, frequency and duration of behaviors (measured in seconds) were identified and noted related to each bird. The study was initiated in March 2007, during four non-consecutive weeks. The observed behaviors were: opening wings, stretching, threatening, ruffling feathers, drinking water, aggressive pecking, eating, running, lying down, stretching head out of the cage, preening, mounting, prostrating, and doing nothing (inactivity). Due to the non-normality of the data recorded, the Kruskal-Wallis statistical test of the MINITAB Statistical Software® was used to compare the medians of the variables. For breed factor, only the durations of the eating presented significant differences (p-value< 0.05). For cage density, there was a significant median difference (p-value< 0.05) for almost all behaviors observed. The average length of time of behaviors was higher for the lowest cage density. However, the frequency of behaviors was lmerfor the lowest cage density. The frequency of the behaviors to preen feathers, to lie down, to drink water and to stretch the head were higher in the aviary, where the groups of birds were smaller. The observed behaviors were particularly affected by experimental factors cage density, and aviary type, which directly affects the available space for each bird.

Carotenóides totais em Pothomorphe umbellata (L.) Miq. cultivadas in vitro

The aim of the present work was to compare the content of carotenoids between callus and regenerated plants of Pothomorphe umbellate. Germinated seeds (40 days old) were inoculated in different concentrations and combinations of BAP (benzylaminopurine) and NAA (naphthalene acetic acid) in order to stimulate the callus' production. After 60 days of culture, the callus containing some shoots were transferred to organogenesis medium (GA 3 0.1 mg L -1, BAP 0.5 mg L -1) for 40 days. Next, they were subcultivated in a medium for seedling growth (without regulators) for 40 days. Callus (collected after 60 days) and seedlings (collected after 140 days) were frozen in liquid nitrogen and kept under -80°C for future carotenoids' analysis. The highest concentration of carotenoids was found in plants cultivated in medium without regulators. The callus did not showed difference concerning the culture medium; however, they presented lower content of carotenoids in relation to plants.

Porous hydroxyapatite scaffolds by polymer sponge method

This study aimed to develop porous hydroxyapatite scaffold for bone regeneration using the replica of the polymeric sponge technique. Polyurethane sponges were used with varying densities to obtain the scaffolds. The results indicate the porous HA scaffolds developed in this study as potential materials for application as bone substitutes to have high porosity (> 70%), chemical composition, interconnectivity and pore sizes appropriate to the bone regeneration.

Eficiência do protocolo Ovsynch em ovelhas da raça Santa Inês

Twenty six Santa Inês ewes were asigned to three treatments to evaluate the efficiency of the Ovsynch protocol. In the treatment 1 - control (n=8), the estrus was synchronized with sponges containing 60 mg MAP for 14 days. On D14, 300 IU eCG were administered. In treatment 2 (n=9) the Ovsynch protocol was used: 25 μg of GnRH (D0), 37.5 μg of PGF2α (D7) and 25 μg of GnRH (D9). In treatment 3 (n=9) the modified Ovsynch protocol was used: the administration of PGF2α and second GnRH as two days early. Estrus detection was accomplished using teaser. All ewes were mated twice with 12 hours of interval. Pregnancy rate (PR) was evaluated by ultrasonography 30 days after the end of mating. Estrus response was of 88.46% on average, and without differences among treatments (p>0.05). The interval for onset of estrus was greater (p<0.05) in T1, when compared with T2 and T3 (41.31±7.2, 13.37±8.42 and 6.75±5.2 h, respectively). The time of receptivity was greater (p<0.05) in females of T1 and T3 (40.5±6.49 h and 53.68±10.27 h, respectively), compared to females of T2 (34.56±7.2 h). The duration of the induced estrus did not differ significant among treatments (p>0.05). Pregnancy rate was significantly greater (p<0.05) in ewes synchronized with Ovsynch protocol (37.5%, 62.5% and 25% for treatment 1, 2 and 3, respectively). The results show superior efficiency of the Ovsynch protocol, under the experimental conditions.

Relações materno-filiais e sua influência no peso pré-desmama de animais Nelore da Bahia

The aim of this study was to investigate the maternal-offspring relationships in the bovine breeds Nellore PO (Pure Breed) and Nellore CL (Cara Limpa) in Bahia State. The genetic composition of breeds did not affect LP (latency time to stand); this variable showed mean values of 71 min and 52 min, respectively, for Nellore PO and Nellore CL. The variable LPTM (latency time to begin suckling after standing) was significantly higher for Nellore PO (mean values of 39 min) tham for Nellore CL (18 min). The variable TCC (time in contact with the calf) was classified as class 4 (60 to 80% of the time was spent with the calves). The P120 (weight at 120 days) had mean values of 122.3 kg and 122.7 kg respectively, for Nellore PO and Nellore CL. In conclusion, the maternal-offspring behaviour had little influence of the genetic composition of the breeds. These results suggest that more studies are necessary to define technological parameters for cattle raising in Bahia State.

On the consistency conditions to braneworlds in scalar-tensor gravity for arbitrary dimensions

We derive an one-parameter family of consistency conditions to braneworlds in the Brans-Dicke gravity. The General Relativity case is recovered by taking a correct limit of the Brans-Dicke parameter. We show that it is possible to build a multiple AdS brane scenario in a six-dimensional bulk only if the brane tensions are negative. Besides, in the five-dimensional case, it is showed that no fine tuning is necessary between the bulk cosmological constant and the brane tensions, in contrast to the Randall-Sundrum model. Copyright © owned by the author(s) under the terms of the Creative Commons Attribution-NonCommercial- ShareAlike Licence.

Exogenous enzymes in pre-starter broiler diets based on corn and soybean meal

A fibrolytic enzyme complex was added to the pre-starter diet. Broiler chicks were randomly distributed into five treatments, consisting of a diet with no enzyme addition and four test diets supplemented with 100, 200, 300 and 400g/T of an enzyme complex. The dietary inclusion of the enzyme complex increased weight gain, and the dose of 300g/T improved weight gain and worsened feed conversion ratio.

Composição Química e Características Fermentativas de Silagens de Cana-de-Açúcar Contendo Farelo de Babaçu

The objective of this trial was to evaluate the effect of babassu meal addition on chemical composition and fermentative characteristics of sugar cane silages. The addition of three concentrations of babassu meal (0; 15 and 30% in fresh matter) was evaluated in a completely randomized design with four replicates. As experimental silos 12 plastic buckets were used, provided with filter closing to allow the exit of fermentation gases. After 45 days, the silos were opened and samples were taken for the determination of dry matter (MS), crude protein (PB), neutral and acid detergent fibers (FDN, FDA) and hemicelluloses (Hem) content, pH values, total acidity quantification (ATT), losses by gases (PG) and effluents (PE). The addition of babassu meal increased the MS contents (38.4% e 40.6%), PB (7.8 e 8.9%), FDN (68.8% e 70.9%) and FDA (43.0% and 45.1%), respectively, with the addition of 15 and 30% of the by-product. The pH values, averaged 4.4, were not affected by the addition of babassu meal. The percentage of ATT increased 89.2%. PG and PE were reduced in 44.8 and 58.8%, respectively. The addition of babassu meal have contributed to the improvement of fermentative parameters and the concentration of CP in the sugar cane silages. However, its use as additive becomes limited due to the increase in fiber content that provides to the silages, depreciating its nutritive value.