Abstract The use of β-galactosidase in food products has been a major focus of the industry. Therefore, the development of efficient and inexpensive methodologies to purify it is essential. Thus, this study aimed to recover the enzyme β-galactosidase (β-gal) by ion-exchange chromatography in a fixed-bed column. Batch adsorption tests were performed using four types of adsorbents. The β-gal adsorption capacity in batch mode using Streamline DEAE resin presented the best performance, with a retention capacity of 18.77 ± 0.14 U/g at pH 6.0. A 22 experimental design was applied to optimize the β-gal recovery using an AKTA Start system, evaluating the ionic strength and the pH as process parameters. The results showed that ionic strength exerted a greater influence on fold purification (FP). The β-gal fraction in elution using 0.1-0.4 M of NaCl showed a yield of 51.65 ± 0.17% and FP of 2.00 ± 0.43. Electrophoresis confirmed the β-gal recovery, where an evident band with a molecular weight between 60 and 120 kDa was observed. These results point to the recovery of a stable β-gal of K. lactis with potential industrial applications.

Abstract: The present review focuses on the proteins and peptides at the interfaces of nanostructured metals and semiconductors as a result of their use in synthesis in-situ and functionalization of nanostructures. We start the review with an introduction about the peculiar properties of nanostructured materials and their applications. In the following, the chemical and structural properties of peptides and proteins that allow their use as reducing, stabilizing, and functionalization agents are discussed. Proteins and peptides have not only the chemical groups for the metal ion reducing but also provide templates for directing the crystalline growing of nanostructures to the desired shapes and sizes. Proteins and peptides are also used mainly for the stabilization and functionalization of a diversity of nanostructured materials providing properties such as biocompatibility, plasmon-enhanced catalysis, sensing, micro/nanomotors, spin filters, and others. Nanostructured materials of metal oxides have mainly been functionalized with proteins and peptides to gain specific properties such as light harvesting and spin filters. Herein, we described the synthesis and functionalization of some types of nanostructured materials by using peptides and proteins. In the last part of the review, it is discussed the perspectives and challenges for the use of proteins and peptides in Nanotechnology.

RESUMO O objetivo deste estudo foi avaliar o efeito da suplementação com diferentes concentrações de glutationa reduzida (GSH - 0; 5; 7,5; 10mM) para criopreservação em cães com avaliações realizadas após glicerolização (refrigeração) e descongelação. Para tal, foram utilizados oito cães e foram realizadas duas coletas de sêmen em intervalo semanal, totalizando 16 amostras de sêmen. Foram avaliadas a motilidade espermática computadorizada (CASA) e a análise de citometria de fluxo do potencial mitocondrial (sonda JC-1) e integridade da membrana/acrossomal (sonda FITC-PI). Subjetivamente foi avaliada a integridade da membrana plasmática e do acrossomal, atividade mitocondrial e integridade do DNA. O plasma seminal foi avaliado quanto à peroxidação lipídica (concentração de TBARS). As amostras refrigeradas e descongeladas suplementadas com 7,5 e 10mM de GSH apresentaram menor porcentagem de espermatozoides com alta atividade mitocondrial (DAB - Classe I) e média (DAB - Classe II). Na concentração de 10mM de GSH, apresentaram maior porcentagem de baixa atividade mitocondrial (DAB - Classe III). Além disso, amostras descongeladas de 10mM de GSH apresentaram taxas de fragmentação de DNA elevadas, provavelmente por efeito de estresse redutivo sobre as mitocôndrias que elevam as espécies reativas de oxigênio e disfunção mitocondrial.

ABSTRACT The aim of this study was to evaluate the effect of supplementation with different concentrations of reduced glutathione GSH (0; 5; 7.5; 10mM) in the extender for cryopreservation in dogs with evaluations performed after glycerolization (chilled) and thawing (thawed). For this purpose, we used 8 dogs and two semen collections were performed in a weekly interval, totaling 16 semen samples. The sperm were analyzed by automatic sperm motility (CASA) and flow cytometry analysis of mitochondrial potential (JC1 dye) and membrane/acrosome integrity (FITC-PI dyes). We evaluated subjectively the membrane and acrosome integrity, mitochondrial activity and DNA integrity. Seminal plasma was evaluated for lipid peroxidation (TBARS concentration). Chilled and thawed samples supplemented with 7.5 and 10mM of GSH had lower percentage of sperm with high (DAB - Class I) and medium (DAB - Class II) mitochondrial activity. And 10mM of GSH had higher percentage of low mitochondrial activity (DAB - Class III). Moreover, thawed samples of 10mM of GSH had high DNA fragmentation rates. Probably by a reductive stress effect on mitochondria which lead to an increase in reactive oxygen species, and a mitochondrial malfunction.

RESUMO O uso de plantas indicadoras pode ser uma alternativa eficiente no monitoramento da presença de moléculas tóxicas no ar, como os herbicidas. Dessa forma, objetivou-se nesta pesquisa avaliar a sensibilidade de plantas forrageiras e daninhas a concentrações residuais atmosféricas de clomazone. Os tratamentos foram arranjados em esquema fatorial 6 x 5, com o primeiro fator correspondendo às espécies vegetais triticale (Triticosecale rimpaui), milho (Zea mays), sorgo (Sorghum bicolor), braquiarão (Urochloa brizantha), beldroega (Portulaca oleracea) e capim-braquiária (Urochloa decumbens), e o segundo, às doses de clomazone de 0, 90, 180, 270 e 360 g ha-1 (equivalentes às concentrações atmosféricas de 0,0, 0,05, 0,10, 0,15 e 0,20 mg L-1). Doze dias após emergência, as plantas foram alocadas no interior de câmaras experimentais retangulares, com volume de 500 dm3, recobertas por filme de polietileno transparente de 150 mm. As plantas ficaram exclusivamente sob a atmosfera da câmara com o herbicida por período de 96 horas. Posteriormente, as câmaras foram abertas e procedeu-se à primeira avaliação da intoxicação e, também, do teor de clorofila. As avaliações foram repetidas aos 7 e 14 dias após abertura da câmara. A presença de clomazone em concentrações mínimas na atmosfera afetou a qualidade das plantas avaliadas. Capim-braquiária, braquiarão, sorgo, triticale e beldroega são espécies sensíveis às concentrações residuais atmosféricas de clomazone, podendo ser utilizadas no monitoramento da qualidade do ar quando há resíduos dessa molécula.

ABSTRACT The use of indicator plants can be an effective alternative in monitoring the presence of toxic molecules in the air, such as herbicides. Thus, in the goal of this study is to assess the sensitivity of forage plants and weeds to atmospheric residual concentrations of clomazone. The treatments were arranged in a 6x5 factorial scheme, with the first factor corresponding to the plant species triticale (Triticosecale rimpaui), maize (Zea mays), sorghum (Sorghum bicolor), palisade grass (Urochloa brizantha), purslane (Portulaca oleracea) and signal grass (Urochloa decumbens), and the second factor to doses of 0, 90, 180, 270 and 360 g ha-1 clomazone doses (equivalent to atmospheric concentrations of 0.0, 0.05, 0.10, 0.15 and 0, 20 mg L-1). Twelve days after emergence, the plants were allocated inside experimental rectangular chambers with a volume of 500 dm3, covered by 150 uM of transparent polyethylene film. The plants remained exclusively in the chamber atmosphere with the herbicide for a period of 96 hours. After the chambers were opened, there were the first evaluation of intoxication and chlorophyll content, as well. The evaluations were repeated on day 7 and 14 after the chamber opening. The presence of clomazone in minimal concentrations in the atmosphere affected the quality of the evaluated plants. Signal grass, palisade grass, sorghum, triticale and purslane were sensitive to atmospheric residual concentrations of clomazone and they can be used to monitor the air quality when there are wastes from this molecule.

RESUMO Relata-se o caso de um cão mestiço, com diagnóstico sorológico para brucelose canina, a partir do qual foram realizadas análises do sêmen epididimário. As amostras espermáticas foram coletadas dos diferentes segmentos epididimários (cabeça, corpo e cauda). Foram realizadas as avaliações de motilidade computadorizada do sêmen (CASA), morfologia espermática, atividade mitocondrial, integridade das membranas plasmática e acrossomal. Houve alteração no padrão de movimentação espermática (motilidade progressiva, espermatozoides rápidos, velocidade média da trajetória, velocidade curvilínea, velocidade linear progressiva, amplitude de deslocamento lateral da cabeça, retilinearidade e linearidade), aumento do total de defeitos morfológicos (51%) e da ausência de atividade mitocondrial espermática (20%) dos espermatozoides, especialmente da cauda do epidídimo. Ressalta-se que tais achados podem contribuir para o diagnóstico clínico da brucelose canina e para a utilização do sêmen epididimário em biotecnologias da reprodução.

ABSTRACT The present work reports a clinical case of a mongrel dog, with serological diagnosis of brucellosis, from which epididymal sperm analysis was performed. Sperm samples were collected from different segments of the epididymis (tail, corpus, and caput). Sperm samples were evaluated for computer-assisted motility analysis (CASA), spermatic morphology, mitochondrial activity and sperm plasmatic membrane and acrosomal integrity. Changes in sperm movement patterns were found (progressive motility, percentage of rapid sperm, percentage of rapid velocity, average pathway, curvilinear velocity, velocity straight line, amplitude of lateral head displacement, straightness and linearity), increase of total morphological defects (51%) and absence of sperm mitochondrial activity (20%) were verified, especially for cauda epididymides. We highlight that such changes can contribute to clinical diagnosis of Brucellosis in dogs and to the use of epididymal sperm in reproductive biotechnologies.

Undernutrition represents a major public health challenge for middle- and low-income countries. This study aimed to evaluate whether a multideficient Northeast Brazil regional basic diet (RBD) induces acute morphological and functional changes in the ileum of mice. Swiss mice (∼25 g) were allocated into two groups: i) control mice were fed a standard diet and II) undernourished mice were fed the RBD. After 7 days, mice were killed and the ileum collected for evaluation of electrophysiological parameters (Ussing chambers), transcription (RT-qPCR) and protein expression (western blotting) of intestinal transporters and tight junctions. Body weight gain was significantly decreased in the undernourished group, which also showed decreased crypt depth but no alterations in villus height. Electrophysiology measurements showed a reduced basal short circuit current (Isc) in the undernourished group, with no differences in transepithelial resistance. Specific substrate-evoked Isc related to affinity and efficacy (glutamine and alanyl-glutamine) were not different between groups, except for the maximum Isc (efficacy) induced by glucose. Transcription of Sglt1 and Pept1 was significantly higher in the undernourished group, while SN-2 transcription was decreased. No changes were found in transcription of CAT-1 and CFTR, while claudin-2 and occludin transcriptions were significantly increased in the undernourished group. Despite mRNA changes, SGLT-1, PEPT-1, claudin-2 and occludin protein expression showed no difference between groups. These results demonstrate early effects of the RBD on mice, which include reduced body weight and crypt depth in the absence of significant alterations to villus morphology, intestinal transporters and tight junction expression.

The objective of this study was to validate the technical skills of nurses in order to provide subsidies for creation of nurse course curriculums for the Nursing School of Federal University of Minas Gerais, Brazil. To this end, we used the Developing a Curriculum method (DACUM), which sets up as a qualitative research technique, based on the creation of a focus group, and that allows identifying duties, skills, attitudes and knowledge of a particular job. This study was conducted in 2005, with 18 nurses from various specialties having the following inclusion criteria: Having 5 to 10 years of professional experience in nursing, being employed solely in a specific job and being considered efficient by other professionals. The results showed areas of nursing with a higher tendency of growth in the labor market, characterized the profile of nurses who work in the health services of Belo Horizonte, and identified the skills required of nurses in various specialized areas. This methodology allowed the development of a collective pedagogic project with the participation of teachers, students and nurses, integrating teaching, service and school.

Neste estudo buscou-se validar as competências técnicas dos enfermeiros a fim de fornecer subsídios para construção do currículo do curso de enfermagem da Escola de Enfermagem da Universidade Federal de Minas Gerais. Para tal, foi utilizado o método Developing a Curriculum (DACUM), que se configura como uma técnica de abordagem qualitativa, baseada na realização de um grupo focal, e que permite o levantamento de funções, habilidades, atitudes e conhecimentos de um determinado cargo. Este estudo foi realizado no ano de 2005 e contou com a participação de 18 enfermeiros de diferentes especialidades que obedeceram aos seguintes critérios inclusão: ter de 5 a 10 anos de experiência profissional na enfermagem, estar empregado exclusivamente no cargo e ser considerado eficiente por seus pares. Como resultados pode-se perceber as áreas de enfermagem com maior tendência de crescimento no mercado de trabalho, caracterizamos o perfil dos enfermeiros que atuam nos serviços de Belo Horizonte e identificamos as competências necessárias para o enfermeiro em diversas áreas de atuação. Esta metodologia permitiu construir um Projeto Pedagógico de forma coletiva com a participação de professores, alunos e enfermeiros dos serviços integrando o ensino, o serviço e a escola.

El objetivo del presente estudio fue validar las competencias técnicas de los enfermeros para proporcionar ayudas para la construcción del plan de estudios de enseñanza del curso de enfermería de la Escuela de Enfermería de la Universidad Federal de Minas Gerais. Con este fin, hemos utilizado el método Developing a Curriculum (DACUM), que se configura como una técnica de enfoque cualitativo, por medio de la realización de un grupo focal, y que permite el levantamiento de las funciones, habilidades, actitudes y conocimientos de un trabajo en particular. Este estudio se llevó a cabo en 2005, con la participación de 18 enfermeras de diferentes especialidades que reúnen los siguientes criterios de inclusión: Tener 5 a 10 años de experiencia profesional en enfermería, estar empleadas exclusivamente en una función y ser considerada eficaz por otros profesionales. Como resultados percibimos las áreas de enfermería con una mayor tendencia de crecimiento en el mercado laboral, la caracterización del perfil de los enfermeros que trabajan en los servicios de Belo Horizonte y la identificación de las habilidades necesarias para la enfermera en diversas áreas de especialización. Esta metodología permitió desarrollar un proyecto de enseñanza de forma colectiva con la participación de profesores, estudiantes y enfermeras de los servicios integrando el servicio, la escuela y la enseñanza.

Nosocomial infections (NI) are frequent events with potentially lethal outcomes. We identified predictive factors for mortality related to NI and developed an algorithm for predicting that risk in order to improve hospital epidemiology and healthcare quality programs. We made a prospective cohort NI surveillance of all acute-care patients according to the National Nosocomial Infections Surveillance System guidelines since 1992, applying the Centers for Disease Control and Prevention 1988 definitions adapted to a Brazilian pediatric hospital. Thirty-eight deaths considered to be related to NI were analyzed as the outcome variable for 754 patients with NI, whose survival time was taken into consideration. The predictive factors for mortality related to NI (p < 0.05 in the Cox regression model) were: invasive procedures and use of two or more antibiotics. The mean survival time was significantly shorter (p < 0.05 with the Kaplan-Meier method) for patients who suffered invasive procedures and for those who received two or more antibiotics. Applying a tree-structured survival analysis (TSSA), two groups with high mortality rates were identified: one group with time from admission to the first NI less than 11 days, received two or more antibiotics and suffered invasive procedures; the other group had the first NI between 12 and 22 days after admission and was subjected to invasive procedures. The possible modifiable factors to prevent mortality involve invasive devices and antibiotics. The TSSA approach is helpful to identify combinations of predictors and to guide protective actions to be taken in continuous-quality-improvement programs.

Matrix metalloproteinases (MMP) are considered to be key initiators of collagen degradation, thus contributing to bone resorption in inflammatory diseases. We determined whether subantimicrobial doses of doxycycline (DX) (<=10 mg kg-1 day-1), a known MMP inhibitor, could inhibit bone resorption in an experimental periodontitis model. Thirty male Wistar rats (180-200 g) were subjected to placement of a nylon thread ligature around the maxillary molars and sacrificed after 7 days. Alveolar bone loss (ABL) was measured macroscopically in one hemiarcade and the contralateral hemiarcade was processed for histopathologic analysis. Groups of six animals each were treated with DX (2.5, 5 or 10 mg kg-1 day-1, sc, 7 days) and compared to nontreated (NT) rats. NT rats displayed significant ABL, severe mononuclear cell influx and increase in osteoclast numbers, which were significantly reduced by 5 or 10 mg kg-1 day-1 DX. These data show that DX inhibits inflammatory bone resorption in a manner that is independent of its antimicrobial properties.

Sete carneiros fistulados no rúmen e no duodeno foram alojados em gaiolas metabólicas e alimentados com feno de Stylosanthes guianensis à vontade. Foi empregada a técnica de sacos de náilon para determinação da degradabilidade in situ do feno, utilizando-se os tempos de 3, 6, 12, 24, 48 e 72 horas para as retiradas dos sacos do rúmen. A taxa de passagem dos sólidos foi determinada utilizando-se cromo mordante como indicador. Os valores de pH no líquido ruminal foram medidos nos tempos de 0, 2, 4, 6 e 8 horas após a alimentação e a concentração de amônia nos tempos de 0, 1, 3, 5, 7, 9 e 11 horas após a alimentação. A taxa de degradação da matéria seca (MS) foi de 8,5%/h, a degradabilidade potencial 38,1% e a degradabilidade efetiva 30,3%. A taxa de degradação da proteína bruta (PB) foi de 9,7%/h, a degradabilidade potencial 56,0% e a degradabilidade efetiva 47,5%. A celulose apresentou maior degradabilidade efetiva que a hemicelulose, com valores de 22,5 e 8,9%, respectivamente. A taxa de passagem dos sólidos foi 2,7%/h. O pH diminuiu linearmente à medida que os tempos de coleta aumentaram. Para o tempo de 5,13 horas após a alimentação, foi estimada a concentração máxima de amônia de 12,18mg/100ml. O feno de S. guianensis apresentou alta taxa de degradação e baixa degradabilidade ruminal da MS e PB.

Seven rumen and duodenal cannulated lambs, were allocated in metabolic cages and were fed ad libitum with Stylosanthes guianensis hay. The in situ technique was used for determination of the degradability of the hay, at 3, 6, 12, 24, 48 and 72 hours of incubation. The passage rate of solids was determined using chromium mordant as external marker. The pH of the rumen liquid was measured at 0, 2, 4, 6 and 8 hours after feeding and the ammonia concentration at 0, 1, 3, 5, 7, 9 and 11 hours after feeding. The degradation rate, the potential degradability, and the effective degradability of dry matter (DM) were 8.5%/h, 38.1% and 30.3%, respectively. The degradation rate of crude protein (CP) was 9.7%/h, the potential degradability was 56.0% and the effective degradabilty was 47.5%. The cellulose presented larger effective degradability than hemicellulose, with the respective values of 22.5% and 8.9%. The passage rate for solids was 2.7%/h. The pH decreased linearly with time of rumen fluid sampling. For the time of 5.13 hours after feeding, the highest ammonia concentration (12.18 mg/100ml) was observed. The S. guinanensis hay showed high degradation rates and low ruminal degradability of DM and CP.

Chemotherapy with oxazaphosphorines, such as cyclophosphamide (CYP), is often limited by unacceptable urotoxicity. Without uroprotection, hemorrhagic cystitis (HC) becomes dose-limiting. To compare the uroprotective efficacy of classical 2-mercaptoethanesulfonic acid (Mesna) treatment with dexamethasone in CYP-induced HC, male Wistar rats (150-200 g; N = 6 in each group) were treated with saline or Mesna (40 mg/kg, ip) immediately and 4 and 8 h after ip administration of CYP (200 mg/kg). One, 2 or 3 doses of Mesna were replaced with dexamethasone (1 mg/kg, ip). The animals were sacrificed 24 h later. Cystitis was evaluated by determining the changes in bladder wet weight (BWW) and by macroscopic and microscopic analysis. CYP treatment induced a marked increased in BWW (162%, P<0.05), which was significantly inhibited by treatment with 3 doses of Mesna (P<0.05; 80%). The replacement of 1 or 2 doses of Mesna with dexamethasone reduced the increase in BWW by 83.3 and 95%, respectively. Macroscopic analysis of the bladder of rats with CYP-induced HC showed severe edema and hemorrhage, confirmed by microscopic analysis, that also showed mucosal erosion, inflammatory cell infiltration and ulcerations. The replacement of 1 or 2 doses of Mesna with dexamethasone inhibited the CYP-induced increase in BWW and almost abolished the macroscopic and microscopic alterations, with no significant difference between the effects of Mesna and dexamethasone, indicating that both drugs were efficient in blocking HC. However, although the replacement of all Mesna doses with dexamethasone reduced the edema, it did not prevent HC, suggesting that Mesna is necessary for the initial uroprotection.