Production, purification and characterization of an extracellular inulinase from Kluyveromyces marxianus var. bulgaricus

The yeast Kluyveromyces marxianus var. bulgaricus produced large amounts of extracellular inulinase activity when grown on inulin, sucrose, fructose and glucose as carbon source, This protein has been purified to homogeneity by using successive DEAE-Trisacryl Plus and Superose 6 HR 10/30 columns. The purified enzyme showed a relative molecular weight of 57 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and 77 kDa by gel filtration in Superose 6 HR 10/30, Analysis by SDS-PAGE showed a unique polypeptide band with Coomassie Blue stain and nondenaturing PAGE of the purified enzyme obtained from media with different carbon sources showed the band, too, when stained for glucose oxidase activity, the optimal hydrolysis temperature for sucrose, raffinose and inulin was 55 degrees C and the optimal pH for sucrose was 4.75, the apparent K-m values for sucrose, raffinose and inulin are 4.58, 7.41 and 86.9 mg/ml, respectively, Thin layer chromatography showed that inulinase from K. marxianus var. bulgaricus was capable of hydrolyzing different substrates (sucrose, raffinose and inulin), releasing monosaccharides and oligosaccharides, the results obtained suggest the hypothesis that enzyme production was constitutive.

Motile sperm organelle morphology examination (MSOME): intervariation study of normal sperm and sperm with large nuclear vacuoles

Background: Although the motile sperm organelle morphology examination (MSOME) was developed only as a selection criterion, its application as a method for classifying sperm morphology may represent an improvement in evaluation of semen quality, with potential clinical repercussions. The present study aimed to evaluate individual variations in the motile sperm organelle morphology examination (MSOME) analysis after a time interval.Methods: Two semen samples were obtained from 240 men from an unselected group of couples undergoing infertility investigation and treatment. Mean time interval between the two semen evaluations was 119 +/- 102 days. No clinical or surgical treatment was realized between the two observations. Spermatozoa were analyzed at greater than or equal to 8400 x magnification by inverted microscope equipped with DIC/Nomarski differential interference contrast optics. At least 200 motile spermatozoa per semen sample were evaluated and percentages of normal spermatozoa and spermatozoa with large nuclear vacuoles (LNV/one or more vacuoles occupying >50% of the sperm nuclear area) were determined. A spermatozoon was classified as morphologically normal when it exhibited a normal nucleus (smooth, symmetric and oval nucleus, width 3.28 +/- 0.20 mu m, length 4.75 +/- 0.20 mu m/absence of vacuoles occupying >4% of nuclear area) as well as acrosome, post-acrosomal lamina, neck and tail, besides not presenting cytoplasm around the head. One examiner, blinded to subject identity, performed the entire study.Results: Mean percentages of morphologically normal and LNV spermatozoa were identical in the two MSOME analyses (1.6 +/- 2.2% vs. 1.6 +/- 2.1% P = 0.83 and 25.2 +/- 19.2% vs. 26.1 +/- 19.0% P = 0.31, respectively). Regression analysis between the two samples revealed significant positive correlation for morphologically normal and for LNV spermatozoa (r = 0.57 95% CI: 0.47-0.65 P < 0.0001 and r = 0.50 95% CI: 0.38-0.58 P < 0.0001, respectively).Conclusions: The significant positive correlation and absence of differences between two sperm samples evaluated after a time interval with respect to normal morphology and LNV spermatozoa indicated that MSOME seems reliable (at least for these two specific sperm forms) for analyzing semen. The present result supports the future use of MSOME as a routine method for semen analysis.

Efficacy of hyaluronic acid binding assay in selecting motile spermatozoa with normal morphology at high magnification

Background: The present study aimed to evaluate the efficacy of the hyaluronic acid (HA) binding assay in the selection of motile spermatozoa with normal morphology at high magnification (8400x).Methods: A total of 16592 prepared spermatozoa were selected and classified into two groups: Group I, spermatozoa which presented their head attached to an HA substance (HA-bound sperm), and Group II, those spermatozoa that did not attach to the HA substance (HA-unbound sperm). HA-bound and HA-unbound spermatozoa were evaluated according to the following sperm forms: 1-Normal morphology: normal nucleus (smooth, symmetric and oval configuration, length: 4.75+/-2.8 mu m and width: 3.28+/-0.20 mu m, no extrusion or invagination and no vacuoles occupied more than 4% of the nuclear area) as well as acrosome, post-acrosomal lamina, neck, tail, besides not presenting a cytoplasmic droplet or cytoplasm around the head; 2-Abnormalities of nuclear form (a-Large/small; b-Wide/narrow; c-Regional disorder); 3-Abnormalities of nuclear chromatin content (a-Vacuoles: occupy >4% to 50% of the nuclear area and b-Large vacuoles: occupy >50% of the nuclear area) using a high magnification (8400x) microscopy system.Results: No significant differences were obtained with respect to sperm morphological forms and the groups HA-bound and HA-unbound. 1-Normal morphology: HA-bound 2.7% and HA-unbound 2.5% (P = 0.56). 2-Abnormalities of nuclear form: a-Large/small: HA-bound 1.6% vs. HA-unbound 1.6% (P = 0.63); b-Wide/narrow: HA-bound 3.1% vs. HA-unbound 2.7% (P = 0.13); c-Regional disorders: HA-bound 4.7% vs. HA-unbound 4.4% (P = 0.34). 3. Abnormalities of nuclear chromatin content: a-Vacuoles >4% to 50%: HA-bound 72.2% vs. HA-unbound 72.5% (P = 0.74); b-Large vacuoles: HA-bound 15.7% vs. HA-unbound 16.3% (P = 0.36).Conclusions: The findings suggest that HA binding assay has limited efficacy in selecting motile spermatozoa with normal morphology at high magnification.

Bond strength between acrylic resin and maxillofacial silicone

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Relative growth and morphological sexual maturity of Uca Cumulanta (Crustacea : Decapoda : Ocypodidae) from a tropical Brazilian mangrove population

The relative growth of the fiddler crab Uca cumulanta was studied, primarily to determine the size at the onset of sexual maturity for a mangrove population in the estuary of the Patitiba River, Paraty, Rio de Janeiro state, Brazil. The evaluation of the morphological sexual maturity of U. cumulanta was performed using the allometric technique. The relationships that most precisely indicated the size at onset of sexual maturity were carapace length (CL) vs propodus length for males and CL vs abdomen width for females. Males and females are mature at 5.25 and 4.75 mm CL, respectively. The remarkable ontogenetic changes observed in the allometric growth of the male major cheliped and the female abdomen, indicate that growth of these structures is closely connected to the timing of sexual maturity. The relative size at onset maturity obtained for this species was 0.68 and this index was compared to that seen in other species in the genus.

Aspectos biológicos de Aphis gossypii Glover, 1877 (Hemiptera: Aphididae) em três cultivares de algodoeiro e em três espécies de plantas daninhas

Este trabalho teve como objetivo estudar os aspectos biológicos de A. gossypii em três cultivares de algodoeiro e em três espécies de plantas daninhas. O experimento foi conduzido no Laboratório de Controle Biológico, sob condições controladas de temperatura (25 ± 1ºC), umidade relativa do ar (70 ± 10%) e fotofase (12 horas). Foram utilizados seis tratamentos, correspondendo às cultivares de algodoeiro CNPA ITA 90, Coodetec 402 e Deltaopal e espécies de plantas daninhas: guanxuma (Sida santaremnensis), malva-preta (Sidastrum micranthum) e trapoeraba (Commelina benghalensis). As unidades experimentais (25 repetições) consistiram de placas de Petri contendo ágar-água a 1% solidificado. Nestas placas, os ofídeos foram mantidos individualmente sobre discos foliares (3cm de diâmetro) de seus respectivos tratamentos. Os menores valores, em dias, para a fase ninfal foram obtidos em CNPA ITA 90 (4,39 ± 0,74), Deltaopal (4,45 ± 0,75), Coodetec 402 (4,47 ± 0,75) e trapoeraba (4,75 ± 0,80). O período reprodutivo e a longevidade dos adultos variaram de 8,21 ± 0,85 a 15,52 ± 0,93 e 11,09 ± 0,97 a 24,33 ± 1,59 dias, respectivamente, sobre guanxuma e Deltaopal. As cultivares de algodoeiro são mais favoráveis a A. gossypii que as plantas daninhas e na ausência de plantas de algodoeiro, as plantas daninhas guanxuma, trapoeraba e malva-preta são hospedeiros potenciais para manutenção da população de A. gossypii.

AIDS-related lymphoma in Brazil - Histopathology, immunophenotype, and association with Epstein-Barr virus

The occurrence of malignant lymphoma is an increasingly important cause of morbidity and mortality in AIDS patients. The incidence of AIDS-related lymphoma in some developing countries such as Brazil is increasing as the survival of HIV infection has improved. Although there is a clear association between several types of immunodeficiency related lymphomas and Epstein-Barr virus (EBV), the association of EBV infection in AIDS-related lymphoma in Brazil, where the incidence of AIDS is high, is unknown. Formalin-fixed, paraffin-embedded tissue from 24 cases of AIDS-related lymphoma in Brazil were analyzed for morphologic classification, immunophenotype, and EBV association using in situ hybridization studies with an EBV-EBER1 biotinylated probe. Twenty cases of AIDS-related lymphoma were classified as non-Hodgkin's lymphoma and four cases were Hodgkin's disease. Eleven non-Hodgkin's lymphomas were classified as diffuse large cell type, five cases were small non-cleaved cell, Burkitt-type, and four cases were large cell immunoblastic non-Hodgkin's lymphoma. Eighteen cases were of B-cell phenotype; one was a T-cell lymphoma, and one was classified as null. Epstein-Barr virus (EBV) was demonstrated in the majority of tumor cells of 11 of 20 (55%) of the cases non-Hodgkin's lymphomas and in 3 of 4 (75%) cases of Hodgkin's disease. AIDS-related lymphomas in Brazil are usually of large cell/immunoblastic type, but Hodgkin's disease is also seen. Both non-Hodgkin's lymphoma and Hodgkin's disease are often associated with EBV infection. The non-Hodgkin's lymphoma is predominantly of B-cell phenotype.

Morphometric and histochemical study of the human vocal muscle

The present study was conducted on vocal muscles removed at autopsy Rom adult individuals (10 men and 8 women, aes ranging from 48 to 78 years) with no laryngeal disease. Histologic analysis was performed with hematoxylin and eosin staining, and histochemical analysis was performed by nicotinamide-adenine-dinucleotide tetrazolium reductase, succinate dehydrogenase, and acid and alkaline myofibrillar adenosine triphosphatase reactions. The histochemical reactions showed that the muscle consists of slow-twitch oxidative (SO), fast-twitch glycolytic (FG), and fast-twitch glycolytic oxidative (FOG) fibers distributed in mosaic form. The frequencies of SO, FOG, and FG fibers were 40.50%, 54.75%, and 4.75%, respectively. The higher frequency of SO and FOG oxidative fibers characterizes the muscle as having aerobic metabolism, resistance to fatigue, and fast contraction. The mean minimum diameters were 31.37 mu m for SO fibers and 36.46 mu m for FOG and FG fibers.

Biologia populacional de Imparfinis minutus (Siluriformes, Heptapteridae) na microbacia do Ribeirão Grande, serra da Mantiqueira oriental, Estado de São Paulo

Populational biology of Imparfinis minutus (Siluriformes, Heptapteridae) in the Ribeirao Grande microbasin, eastern Mantiqueira mountain range, São Paulo State. The aim of the work was to analyze biological aspects and distribution of Imparfinis minutus in the Ribeirao Grande microbasin (22[degree]4708[minute]''S, 45[degree]28[minute]17''W), in eastern Mantiqueira mountain range, from July 2001 to April 2002. Among the 37 species sampled, I. minutus (Siluriformes, Heptapteridae), was distributed from the piedmont to the plains of the Vale do Paraiba. The occurrence of this species was associated with the environments of small water volume, bottom with stones and gravel. The amplitude length of I. minutus was 4.75 to 12.75 cm. The length-weight relationship of the population was estimated at W = 0.0052 L3,13. Analyzing the alometric (Ka) and relative (Kr) condition factors, both indexes presented similar values. The lowest value was obtained in July, increasing gradually in October and January, and peaking in April. The analysis of the gonadossomatic relationship (GSR) of females showed higher values in July and January. The reproductive period of I. minutus occurred on spring/summer (October to January), as suggested by the gonadossomatic relationship and condition factor results. Feeding activity was constant, with fat visceral deposition concentrated in April and July. Imparfinis minutus is an aquatic insectivore opportunist species, consuming Diptera and Trichoptera larvae as secondary items. Most ingested food items were autochthonous resources.

Zygomatic bone: Anatomic bases for osseointegrated implant anchorage

Purpose: The aim of the present study was to evaluate zygomatic bone thickness considering a possible relationship between this parameter and cephalic index (Cl) for better use of Cl in the implant placement technique. Materials and Methods: Cl was calculated for 60 dry Brazilian skulls. The zygo matic bones of the skulls were divided into 13 standardized sections for measurement. Bilateral measurements of zygomatic bone thickness were made on dry skulls. Results: Sections 5, 6, 8, and 9 were appropriate for implant anchorage in terms of location. The mean thicknesses of these sections were 6.05 mm for section 5, 3.15 mm for section 6, 6.13 mm for section 8, and 4.75 mm for section 9. In only 1 section, section 8, did mean thickness on 1 side of of the skull differ significantly from mean thickness on the other side (P <.001). Discussion: For the relationship between quadrant thick ness and Cl, sections 6 and 8 varied independently of Cl. Section 5 associated with brachycephaly, and section 9 associated with subbrachycephaly, presented variations in the corresponding thickness. Conclusion: Based on the results, implants should be placed in sections 5 and 8, since they presented the greatest thickness, except in brachycephalic subjects, where thickness was greatest in section 5, and in subbrachycephalic subjects, where thickness was greatest in section 9. Cl did not prove to be an appropriate parameter for evaluating zygomatic bone thickness for this sampling.

Substituição dos grãos secos de milho pela silagem de grãos úmidos de milho para vacas da raça holandesa em lactação

In order to evaluate the effects of replacement of dried corn (GSM) for silage moisture corn (HMCS) on production and milk composition were used five Holstein cows, primiparous with a mean of 112 days post delivery, confined in Tie Stall, for 70 days. We used five diets according to NRC to 17.5% CP (DM) and 2.4 Mcal / kg DM, the 40:60 ratio of roughage and concentrate containing soybean meal, sugar cane silage and hay and substitution levels of the GSM HMCS the following treatments: a) 0%, 2) 25%, 3) 50%, 4) 75% and 5) 100%. Milk production and dry matter intake (DMI) were recorded daily. The animals were milked daily 6:00 and 18:00 h, and milk samples collected from consecutive milkings of each experimental period of 14 days (four days of collection). The experimental design was a 5x5 Latin square and the data analyzed by the statistical program SAS. Body weight (508 kg), milk (23.6 kg), corrected milk (22.7 kg), DMI (17.13 kg) showed no significant difference, but the intakes of neutral detergent fiber (6.67 kg), and detergent acid (3.39 kg), feed efficiency for the production of milk (milk 1.41 kg / day) urea nitrogen (17.67 mg / dL) differ, thus indicating that HMCS is more efficient than GSM in the diet of dairy cows not alter the production and milk composition.

Genetic and non-genetic effects on calf vigor at birth and preweaning mortality in nellore calves

The objective of the present study was to evaluate the genetic and non-genetic effects that influencevigor at birth and preweaning mortality in Nellore calves. A total of 11,727 records of births that occurred between 1978 and 2006, offspring of 363 sires, were analyzed. Poor calf vigor at birth (VB) and preweaning mortality divided into stillbirth (SB), early mortality (EM) and total mortality (TM) were analyzed as binary variables. Generalized linear models were used for the evaluation of non-genetic effects and generalized linear mixed models for genetic effects (sire and animal models). The incidences were 4.75% for VB, 2.66% for SB, 5.28% for EM, and 7.99% for TM. Birth weight was the effect that most influenced the traits studied. Calves weighing less than 22kg(females) and less than 24kg (males) were at a higher risk of low vigor and preweaning mortality. Preweaning mortality was higher among calves born from cows aged .3 and .11 years at calving compared with cows aged 7 to 10 years. Male calves presented less vigor and higher preweaning mortality than female calves. Selection for postweaning weight did not influence preweaning mortality. The heritability estimates ranged between 0.01 and 0.09 for VB, 0.00 and 0.27 for SB, 0.03 and 0.17 for EM and 0.02 and 0.10 for TM. Stillbirth should be included as a selection criterion in breeding programs of Nellore cattle, alone or as part of a selection index, aiming to reduce preweaning mortality. © 2013 Sociedade Brasileira de Zootecnia.

Persistência de palhada de plantas de cobertura em função de doses de silício e resposta do feijoeiro em sucessão

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Programa de exercícios físicos: análise dos fatores de risco cardiovaculares e biomarcadores de inflamação e estresse oxidativo em diabéticas exercitadas e não exercitadas

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Parâmetros cinéticos e têmporo-espaciais de gatos clinicamente saudáveis

Pós-graduação em Medicina Veterinária - FMVZ