Abstract The Agaricomycetes fungi produce various compounds with pharmaceutical, medicinal, cosmetic, environmental and biotechnological properties. In addition, some polysaccharides extracted from the fungal cell wall have antitumor and immunomodulatory actions. The aim of this study was to use genetic modification to transform Schizophyllum commune and identify if the phenotype observed (different from the wild type) resulted in changes of the cell wall polysaccharides. The plasmid pUCHYG-GPDGLS, which contains the Pleurotus ostreatus glucan synthase gene, was used in S. commune transformations. Polysaccharides from cell wall of wild (ScW) and mutants were compared in this study. Polysaccharides from the biomass and culture broth were extracted with hot water. One of the mutants (ScT4) was selected for further studies and, after hydrolysis/acetylation, the GLC analysis showed galactose as the major component in polysaccharide fraction from the mutant and glucose as the major monomer in the wild type. Differences were also found in the elution profiles from HPSEC and NMR analyses. From the monosaccharide composition it was proposed that mannogalactans are components of S. commune cell wall for both, wild and mutant, but in different proportions. To our knowledge, this is the first time that mannogalactans are isolated from S. commune liquid culture.

Abstract Bisphenol A is an endocrine interfering compound, produced and used on a large scale worldwide. Chemical and biologic methods can be used to remove it from the environment. Biological methods are considered less costly, safer and, according to green chemistry definitions, an environmentally correct method. Considering the use of a crude enzyme broth, without any downstream process, the costs could be mostly reduced. Thus, the removal of bisphenol A by Pleurotus sajor-caju crude enzyme broth was investigated. Initially, the agro-industrial wastes were characterized and, the composition of the culture medium and the bioreactor culture conditions were defined. The enzyme produced in the highest concentration was characterized and the crude broth used in the bisphenol A removal assays. The OXI45 culture medium presented the highest laccase activity (1,850.7 U L-1, 350 rpm). Greater laccase stability was observed at 20 - 40 oC and pHs 5 - 7. Vanillin and ferulic acid (considered mediator compounds) were identified in the crude broth, probably helping on the obtention of the high value of removal effectiveness (0.052 mg U-1 h-1). The results indicate the potential use of the Pleurotus sajor-caju crude enzyme broth to obtain an enzymatic formulation for application in the environmental area.

RESUMO Este estudo objetiva reportar a ocorrência de apoptose in vivo induzida pelo veneno da serpente Bothrops alternatus em células musculares esqueléticas. Cinco coelhos machos, adultos, receberam 150μg/kg de veneno no músculo vasto lateral, enquanto outros cinco animais receberam 0,1% de BSA diluído em PBS no mesmo local. Após 12 horas, os animais foram eutanasiados, e amostras do local de inoculação foram coletadas para análise histopatológica. Foram evidenciadas necrose e hemorragia nas células musculares. Além disso, a análise imuno-histoquímica para identificação de caspase-3 ativada revelou marcações granulares e agregadas no citoplasma das células musculares, compatíveis com o processo de apoptose. Este é o primeiro relato que confirma o veneno de B. alternatus como causador de apoptose in vivo em células musculares esqueléticas.

RESUMO: Este estudo objetivou determinar a soroprevalência da Babesiose e Anaplasmose em bovinos dos municípios de Ouricuri e Petrolina, estado de Pernambuco, Brasil; e definir os possíveis fatores de risco para a ocorrência dessas doenças. Amostras de sangue foram coletadas para realização de teste sorológico por Imunofluorescência Indireta (RIFI). Questionários epidemiológicos sanitários foram aplicados aos produtores com o objetivo de identificar possíveis fatores de risco. Carrapatos foram coletados, identificados e testados por Reação em Cadeia da Polimerase (PCR) para o diagnóstico da infecção por Anaplasma marginale, Babesia bigemina e Babaesia bovis. O estudo foi conduzido com 861 bovinos, sendo 468 de Petrolina e 393 de Ouricuri. A soroprevalência de A. marginale, B. bigemina e B. bovis em Petrolina foi de 35,0% (164/468), 35,9% (168/468) e 32,3% (151/468), respectivamente; e em Ouricuri foi de 45,5% (179/393), 38,6% (152/393) e 54,9% (216/393), respectivamente. A co-infecção por Anaplasma spp. e Babesia spp. foi observada em 31,6% e 32,1% de amostras de Petrolina e Ouricuri, respectivamente. A detecção de DNA de Babesia spp. por PCR foi possível em 5,8% (8/137) carrapatos, dos quais em 62,5 % (5/8) foi detectada posteriormente infecção por B. bovis, e em 23,3% (32/137) por A. marginale. A presença de carrapatos, o uso de acaricidas, idade, raça, e o município de residência dos animais foram identificados como fatores de risco para TPB pela análise univariável e multivariável. Este estudo permitiu caracterizar os municípios estudados como de instabilidade enzoótica para esses hemoparasitos, e consequentemente, alertar para adoção de medidas adequadas de controle e realização de novos estudos.

ABSTRACT: This study aimed to determine the seroprevalence of Babesiosis and Anaplasmosis in cattle from the municipalities of Ouricuri and Petrolina, state of Pernambuco, Brazil, and to define the risk factors for the occurrence of the diseases. Blood samples were collected for serologic testing by Indirect Immunofluorescence Assay (IFA). Sanitary epidemiological questionnaires were applied to the producers aiming to identify possible risk factors. Ticks were collected, identified and tested by Polymerase Chain Reaction (PCR) for the diagnosis of infection by Anaplasma marginale, Babesia bigemina and Babesia bovis. The study was conducted with 861 cattle, being 468 in Petrolina and 393 in Ouricuri. The seroprevalence of A. marginale, B. bigemina and B. bovis in Petrolina was of 35.0% (164/468), 35.9% (168/468) and 32.3% (151/468), respectively; and in Ouricuri was 45.5% (179/393), 38.6% (152/393), and 54.9% (216/393), respectively. Co-infection for Anaplasma spp. and Babesia spp. was observed in 31.6% and 32.1% samples of Petrolina and Ouricuri, respectively. The detection of DNA of Babesia spp. by PCR was possible in 5.8% (8/137) ticks; which 62.5% (5/8) was detected later infection with B. bovis; and 23.3% (32/137) with A. marginale. The presence of ticks, the use of acaricide, age, race, and county of residence of the animals were identified as risk factors for TBD by univariate analysis and multivariate. This study allowed the characterization of the municipalities studied as enzootic instability areas for these hemoparasitic, and consequently alert for adoption of adequate control measures and new studies.

ABSTRACT Fungi of Pleurotus genus have attracted a great interest due to their medicinal properties such as anti-inflammatory, antimicrobial and antitumor. These properties are attributed mainly to polysaccharides synthesized by Pleurotus. This work aimed to study the mycelial growth of P. ostreatus in submerged culture, evaluating the influence of the initial concentration of substrate (20 and 40 g/L of glucose) and the pH (4 and 6) on kinetic parameters of production of biomass. The effectiveness of different doses (10, 30 and 50 mg/kg) of a mycelium polysaccharide fraction extracted from P. ostreatus in reducing Sarcoma 180 development in mice was also verified. In the range of this study, maximum concentration of mycelial biomass (about 12.8 g/L) was obtained using 40.0 g/L of glucose, at pH 4.0. The total biomass productivity (Px) was not significantly affected by substrate concentration and pH, reaching values of 0.034 g/L.h. Sarcoma 180 tumor weight was reduced in 74.1, 75.5 and 53.7% when 10, 30 and 50 mg/kg were administered, respectively. These results show the high antitumor potential of intracellular polysaccharide fraction of mycelial biomass of P. ostreatus, particularly at lower doses of 10 and 30 mg/kg.

A leishmaniose visceral é uma enfermidade cujo agente etiológico no Brasil é o protozoário Leishmania infantum chagasi. Os cães são considerados reservatórios urbanos da doença, sendo indicadores da ocorrência de casos humanos. O presente trabalho teve como objetivo diagnosticar a infecção por L. infantum chagasi em cães domiciliados e errantes do município de Belém, estado do Pará, através da reação em cadeia da polimerase (PCR) e da reação de imunofluorescência indireta (RIFI), empregando dois antígenos distintos. Amostras de sangue venoso de cães adultos, sem distinção de sexo ou raça, de diferentes bairros e épocas do ano da cidade de Belém-PA, foram colhidas em tubos sem e com anticoagulante para obtenção do soro e do DNA, respectivamente. Esses animais foram divididos em dois grupos: cães errantes capturados pelo Centro de Controle de Zoonoses (Grupo A) e cães domiciliados (Grupo B). Os soros foram analisados através do teste de RIFI para pesquisa de IgG utilizando-se dois antígenos distintos: 1) antígeno do kit Bio-Manguinhos/FIOCRUZ (Ag-PRO) contendo formas promastigotas de Leishmania sp. (complexo major-like); 2) Antígeno do Instituto Evandro Chagas (Ag-AMA) constituído por formas amastigotas de L. infantum chagasi. A avaliação dos dois antígenos foi realizada com as amostras reagentes a partir da titulação 1:80. Já a PCR foi realizada a partir do DNA extraído do sangue total dos animais e amplificado utilizando-se os iniciadores RV1e RV2. Das 335 amostras analisadas, 10,4% (35/335) foram reagentes na RIFI (Ag-PRO) e 0,9% (3/335) reagiram com o Ag-AMA. A distribuição das amostras positivas se deu da seguinte forma: Grupo A 14,8% (25/169) com Ag-PRO e 1,2% (2/169) com Ag-AMA; Grupo B 6% (10/166) com Ag-PRO e 0,6% (1/166) com Ag-AMA; sendo que todas as amostras positivas pelo teste de RIFI com o Ag-AMA também reagiram com o Ag-PRO e em nenhuma das amostras foi detectado o DNA de L. infantum chagasi. Os achados do presente estudo indicam que Belém ainda pode ser considerada área não endêmica para leishmaniose visceral canina e que a natureza do antígeno influencia no resultado da RIFI para a pesquisa de anticorpos anti-L. infantum chagasi em cães, sendo que a RIFI que utiliza formas promastigotas de Leishmania major-like como antígeno deve ser utilizada com cautela como método diagnóstico confirmatório em estudos epidemiológicos em áreas não endêmicas para LVC.

Visceral leishmaniasis is a disease whose etiological agent in Brazil is Leishmania infantum chagasi. Dogs are considered urban reservoirs of the disease, being an indicator of the human cases occurrence. The present study aimed to diagnose L. infantum chagasi infection in stray and owned dogs in Belém, Pará State, by polymerase chain reaction (PCR) and indirect immunofluorescence assay (IFA) using two different antigens. Venous blood samples from adult dogs, regardless of gender or breed, of different neighborhoods in Belém-PA, were collected in tubes with and without anticoagulant to obtain DNA and serum, respectively. These animals were divided into two groups: stray dogs captured by the Center for Zoonosis Control (Group A) and owned dogs (Group B). Sera were analyzed by IFA testing for IgG using two different antigens: 1) Bio-Manguinhos/Fiocruz antigen kit (Ag-PRO) containing promastigotes of Leishmania sp. (Complex Major-Like), 2) Instituto Evandro Chagas Antigen (Ag-AMA) consisting of amastigotes of L. infantum chagasi. The evaluation of the two antigens was performed considering positive the reactions above the 1:80 dilution. Already PCR was performed with DNA isolated from whole blood of animals and amplified with the primers RV1 and RV2. Of the 335 samples analyzed, 10.4% (35/335) were positive by IFA (Ag-PRO) and 0.9% (3/335) with the Ag-AMA. The distribution of positive samples is given as follows: Group A 14.8% (25/169) with Ag-PRO and 1.2% (2/169) with Ag-AMA; Group B 6% (10/166) with Ag-PRO and 0.6% (1/166) with Ag-AMA, being that all samples positive by IFA with Ag-AMA also reacted with Ag-PRO, and none of the samples detected DNA of L. infantum chagasi. The findings of this study indicate that Belém can still be considered non-endemic area for canine visceral leishmaniasis and the nature of the antigen influences the result of the IFA for the detection of anti-L. infantum chagasi antibodies in dogs, and the IFA using promastigotes of Leishmania major-like antigen should be used with caution as a confirmatory diagnostic on epidemiological studies in non-endemic areas.

Espécies vegetais da família Apiaceae podem proporcionar recursos vitais para insetos predadores de pragas agrícolas, promovendo sua eficiência como agentes de controle biológico devido à oferta de recursos como sítios de refúgio, proteção e alimentação. Assim, o objetivo deste trabalho foi avaliar a atração de insetos predadores pelas apiáceas coentro (Coriandrum sativum L.), endro (Anethum graveolens L.) e funcho (Foeniculum vulgare Mill.). O experimento foi conduzido no campus da Universidade Federal de Lavras (UFLA), Lavras, MG, em blocos completos ao acaso, com três tratamentos e quatro repetições. Coletas semanais foram realizadas por meio de batida das plantas em bandejas, sendo os predadores aspirados e acondicionados em frascos com álcool 70%. Determinaram-se a curva do coletor, a riqueza de espécies, o índice de diversidade de Shannon-Wienner e o índice de similaridade (análise de Cluster). O funcho apresentou maior índice de Shannon-Wienner (H' = 1,104) e riqueza, enquanto o coentro mostrou maior abundância, sobretudo para a espécie Orius insidiosus (Hemiptera: Anthocoridae), um importante predador de tripes em hortaliças. Outras espécies predadoras de pragas de hortaliças foram encontradas, de maneira geral, em todas as apiáceas, destacando-se os adultos e larvas de coccinelídeos, importantes predadores de pulgões. A análise de Cluster indicou maior similaridade entre funcho e endro (55,03%). O período de floração promoveu considerável aumento na abundância de predadores, sobretudo no coentro.

Species of plants of the Apiaceae family can provide vital resources for predators of agricultural pests, boosting their efficiency as biological control agents, because they allow their maintenance in the environment by offering features such as sites of refuge, protection and food. The present study was aimed to evaluate the attraction of predator insects by 3 Apiaceae species, namely coriander (Coriandrum sativum L.), dill (Anethum graveolens L.) and fennel (Foeniculum vulgare Mill.). The bioassay was conducted on the campus of UFLA (MG) in a randomized block design with 3 treatments and 4 replications. Weekly collections were carried out by beating the plants on white plastic trays, aspirating the predators and packing them into bottles with 70% alcohol. Determinations were made of the collector curve, the species richness, the Shannon-Wienner diversity index and the index of similarity (cluster analysis). The fennel had a higher richness and Shannon-Wienner index (H' = 1.104), while the coriander was more abundant, especially for the species Orius insidiosus (Hemiptera: Anthocoridae), a major predator of thrips in vegetables. Other predatory species of vegetable pests were found, generally, on all Apiaceae, especially adults and larvae of coccinellids, which are important predators of aphids. Cluster analysis indicated a greater similarity between fennel and dill (55.03%), and a lower value for the other pairs. The flowering period increased the abundance of predators, especially the coriander.

Foi realizado o fracionamento químico do extrato aquoso da Mascagnia rigida Griseb., uma importante planta tóxica no Brasil, para se obter cinco substratos ricos em diferentes grupos químicos - alcalóides, flavanóides, taninos, saponinas e açúcares, ácidos orgânicos e aminoácidos -, e investigar a toxicidade dessas frações, exceto a última, em 75 camundongos. Os animais, distribuídos aleatoriamente em cinco grupos, receberam: grupo I - alcalóides; grupo II - flavanóides; grupo III - taninos; grupo IV - saponinas e grupo V - placebo, este último funcionando como controle negativo. Todos os grupos, com exceção do grupo-controle, redistribuídos em três subgrupos, A, B e C, com cinco animais cada, receberam, respectivamente, 9g/kg, 18g/kg e 27g/kg de cada substrato. As frações foram fornecidas via oral, diariamente, por sete dias; no sétimo dia, foi coletado sangue para o estudo do perfil sangüíneo e dosagem de enzimas musculares. As frações de alcalóides e taninos foram capazes de causar alteração no perfil enzimático-muscular, com aumento significativo da enzima miocárdica. Observou-se, também, aumento significativo na porcentagem da CK-MB após a administração das frações de saponinas e taninos, comprovando a ação tóxica da M. rigida sobre a fibra muscular cardíaca.

A study was carried out to examine the toxic effects of Mascagnia rigida Griseb. Four classes of compounds were extracted from the plant by phytochemistry study and the individual effect of each one on mice was examined as follows: group I - alkaloids; group II- flavones; group III - tannins; group IV - saponins; and group V - water (control). Each group was further divided in three subgroups, A (9g/kg), B (18g/kg), and C (27g/kg), with five mice orally receiving a particular dose, once per day for one week. On the 7th day, blood was collected and hematological exams and levels of muscle enzymes were analyzed. The results showed that both alkaloids and tannins caused a significant increase in myocardial enzyme. Administration of either saponins or tannins fractions caused an increase of CK-MB enzyme. This study showed that Mascagnia rigida Griseb has the ability to cause damage to myocardial fibers.

Serum hormone levels were compared between captive and free-living maned wolves and seasonal variations of sex hormones were studied. Blood samples were collected from 16 male and 26 female adult animals from Brazilian zoos, and from 30 male and 24 female free-living adults to determine serum progesterone and testosterone by radioimmunoassay. Serum testosterone concentrations varied (P < 0.05) across seasons for 16 captive males, being higher in autumn (2184.7 ± 355.1 pg/mL) than in summer (1080.7 ± 205.4 pg/mL), winter (1270.1 ± 276.6 pg/mL) and spring (963.9 ± 248.1 pg/mL), although they did not differ between summer, winter and spring. Testosterone concentration of 30 free-living males differed (P < 0.05) between autumn (824.1 ± 512.2 pg/mL), winter (14.4 ± 8.0 pg/mL) and spring (151.9 ± 90.5 pg/mL). Comparison between captive and free-living animals showed no difference in autumn (P > 0.05). Sixteen captive males showed higher testosterone concentration during winter and spring compared with 30 free-living animals (P < 0.05). Progesterone concentration varied among seasons in 26 captive females (P < 0.05), being higher in autumn (15.3 ± 3.1 ng/mL) than in summer (6.6 ± 1.5 ng/mL), winter (5.3 ± 3.1 ng/mL) and spring (4.3 ± 0.7 ng/mL). Progesterone concentration of 24 free-living females varied between autumn (17.1 ± 6.0 ng/mL) and winter (1.7 ± 0.3 ng/mL) (P < 0.05), but we could not obtain data for spring or summer. No difference in progesterone levels was observed between captive and free-living females in autumn and winter.

Avaliamos 31 crianças com mielomeningocele nascidas entre julho de 1990 e julho de 2000. A mediana de acompanhamento foi 24 meses (6-68m). Duas mães possuíam conhecido fator de risco (diabetes mellitus). Doze obtiveram correto diagnóstico pré-natal da patologia. Todas as crianças nasceram a termo; 23 via cesariana; 13 apresentaram ruptura de membrana. A mediana de correção cirúrgica foi 4 dias (1-44d). Lesões lombosacras foram as mais freqüentes (46%). Trinta pacientes apresentaram hidrocefalia, sendo a derivação ventrículo peritoneal (DVP) necessária em 27. Meningite foi 4 vezes mais frequente em pacientes com DVP. Sete pacientes eram epilépticos (19,4%). O teste de Denver II mostrou atraso motor significante. Bexiga neurogênica foi diagnosticada em 12 pacientes. Pé torto congênito foi a malformação ortopédica mais comum (53%). Seis pacientes morreram. Até o momento, 17 pacientes são acompanhados. Uma abordagem multidisciplinar provavelmente colabora para melhor qualidade de vida.

We analyzed 31 children with myelomeningocele born between July 1990 and July 2000. Follow-up median was 24 months (6-68months). Only 2 mothers had a known etiologic factor (diabetes mellitus). Twelve had the correct prenatal diagnosis. All children were born at term; 23 by cesarean; 13 had rupture of the membrane. Surgical correction had a 4 days median (1 to 44 days). Lumbosacral lesions were the most frequent (46%). Thirty patients were hydrocephalic, shunt was placed in 27. Meningitis was 4 times more frequent in shunted patients. Seven became epileptic (19.4%). Denver II test showed significant delay in gross motor development . Neurogenic bladder was diagnosed in 12 patients. Congenital clubfoot was the main orthopedic malformation (53%). Six infants died. Nowadays, 17 patients are being followed. A multidisciplinary approach probably helps for a better quality of life.

The influence of oxygen supply rate on the growth and the production of both ethanol and acetaldehyde by the aerotolerant fermentative bacterium Z. mobilis is discussed in this work. The results showed similar values of cell mass yield (0.043 g/g) for the five different levels of initial volumetric oxygen transfer coefficient (K La) studied. The maximum specific growth rate (µx,m.) under anaerobic conditions was higher than those found in aerated runs. Anaerobic cultivation resulted in the best ethanol yield (0.38 g/g). For initial K La values of 62, 94, and 118 h-1, ethanol yields were 0.10, 0.12, and 0.09 g/g, respectively, whereas for K La of 30 h-1, an intermediate value (0.24 g/g) was achieved. Under anaerobiosis, no acetaldehyde was produced. With initial K La values of 62, 94, and 118 h-1, acetaldehyde yields were similar (0.12 to 0.17 g/g), and for K La of 30 h-1 only 0.07 gram of acetaldehyde was formed per gram of glucose. Although increasing values for the maximal specific acetaldehyde formation rate were calculated as K La was increased, our results showed that the presence of an excess of dissolved oxygen throughout fermentation is enough to provide appropriate conditions for the production of acetaldehyde by Z. mobilis.