Diluent Containing Dimethylformamide Added With Sucrose Improves In Vitro Quality After Freezing/Thawing Stallion Sperm.

The aim of this study was to investigate the effects of sucrose on post-thawed equine semen quality. Semen samples (n = 24) were collected from six stallions. They were diluted (200 × 106 sperm/mL) in a freezing medium based on skimmed milk, egg yolk, dimethylformamide, and supplemented with sucrose at concentrations of 0 (Control), 25, 50, and 100 mM and in a commercial extender (BotuCrio). Subsequently, they were filled in straws (0.5 mL) and subjected to freezing and storage (-196°C). Immediately after thawing (37°C, 30 seconds), semen samples were evaluated for kinetics (CASA), plasma and acrosomal membrane integrity, and mitochondrial membrane potential (flow cytometry). The addition of 50 and 100mM sucrose to the freezing extender increased (P < .05) the parameters of TM, PM, VCL, VSL, and VAP, compared to the control group. The WOB parameter of the group supplemented with 100 mM sucrose was higher (P < .05) than the control group. Higher values ​​(P < .05) of ALH and BCF were observed in groups treated with sucrose (25, 50, and 100 mM), compared to BotuCrio. The semen frozen in the presence of 100 mM sucrose presented higher percentages (P < .05) of sperm with intact plasma and acrosomal membranes, and high mitochondrial membrane potential in relation to the other groups. It is concluded that the addition of sucrose to equine semen freezing extender increase motility (50 and 100 mM), plasma and acrosomal membrane integrity preserve, and high sperm mitochondrial membrane potential (100 mM) after thawing.

Neural Stem Cells and Fetal-Onset Hydrocephalus.

Fetal-onset hydrocephalus is not only a disorder of cerebrospinal fluid (CSF) dynamics, but also a brain disorder. How can we explain the inborn and, so far, irreparable neurological impairment in children born with hydrocephalus? We hypothesize that a cell junction pathology of neural stem cells (NSC) leads to two inseparable phenomena: hydrocephalus and abnormal neurogenesis. All neurons, glial cells, and ependymal cells of the mammalian central nervous system originate from the NSC forming the ventricular zone (VZ) and the neural progenitor cells (NPC) forming the subventricular zone. Several genetic mutations and certain foreign signals all convey into a final common pathway leading to cell junction pathology of NSC and VZ disruption. VZ disruption follows a temporal and spatial pattern; it leads to aqueduct obliteration and hydrocephalus in the cerebral aqueduct, while it results in abnormal neurogenesis in the telencephalon. The disrupted NSC and NPC are released into the CSF and may transform into neurospheres displaying a junctional pathology similar to that of NSC of the disrupted VZ. These cells can then be utilized to investigate molecular alterations underlying the disease and open an avenue into possible NSC therapy.

Understanding How the Subcommissural Organ and Other Periventricular Secretory Structures Contribute via the Cerebrospinal Fluid to Neurogenesis.

The dynamic and molecular composition of the cerebrospinal fluid (CSF) and, consequently, the CSF physiology is much more complex and fascinating than the simplistic view held for decades. Signal molecules either transported from blood to CSF or secreted into the CSF by circumventricular organs and CSF-contacting neurons, use the CSF to reach their targets in the brain, including the pre- and postnatal neurogenic niche. The subcommissural organ (SCO), a highly conserved brain gland present throughout the vertebrate phylum, is one of the sources for signals, as well as the choroid plexus, tanycytes and CSF-contacting neurons. The SCO secretes into the fetal and adult CSF SCO-spondin, transthyretin, and basic fibroblast growth factor. These proteins participate in certain aspects of neurogenesis, such as cell cycle of neural stem cells, neuronal differentiation, and axon pathfinding. Through the CSF, the SCO-secretory proteins may reach virtually any target in the embryonic and adult central nervous system. Since the SCO continues to secrete throughout life span, it seems likely that the neurogenetic property of the SCO compounds would be targeted to the niches where neurogenesis continues in adulthood. This review is aimed to bring into discussion early and new evidence concerning the role(s) of the SCO, and the probable mechanisms by which SCO compounds can readily reach the neurogenic niche of the subventricular zone flowing with the CSF to participate in the regulation of the neurogenic niche. As we unfold the multiples trans-fluid talks between discrete brain domains we will have more tools to influence such talks.

Structure and function of the ependymal barrier and diseases associated with ependyma disruption.

The neuroepithelium is a germinal epithelium containing progenitor cells that produce almost all of the central nervous system cells, including the ependyma. The neuroepithelium and ependyma constitute barriers containing polarized cells covering the embryonic or mature brain ventricles, respectively; therefore, they separate the cerebrospinal fluid that fills cavities from the developing or mature brain parenchyma. As barriers, the neuroepithelium and ependyma play key roles in the central nervous system development processes and physiology. These roles depend on mechanisms related to cell polarity, sensory primary cilia, motile cilia, tight junctions, adherens junctions and gap junctions, machinery for endocytosis and molecule secretion, and water channels. Here, the role of both barriers related to the development of diseases, such as neural tube defects, ciliary dyskinesia, and hydrocephalus, is reviewed.

Antimicrobial, antiproliferative and proapoptotic activities of extract, fractions and isolated compounds from the stem of Erythroxylum caatingae plowman.

In the study, we have examined the antitumor and antimicrobial activities of the methanol extract, the fractions, a fraction of total alkaloids and two alkaloids isolated from the stem of Erythroxylum caatingae Plowman. All test fractions, except the hexane fractions, showed antimicrobial activity on gram-positive bacteria and fungi. The acetate: methanol (95:5), acetate, chloroform and hexane fractions show the highest cytotoxicity activity against the NCI-H292, HEp-2 and K562 cell lines using MTT. The absence of hemolysis in the erythrocytes of mice was observed in these fractions and 6ß-Benzoyloxy-3α-(3,4,5- trimethoxybenzoyloxy) tropane (catuabine B). Staining with Annexin V-FITC and JC-1 was used to verify the mechanism of action of the compounds of E. caatingae that showed cytotoxicity less than 30 µg/mL in leukemic cells. After 48 h of incubation, we observed that the acetate: methanol (95:5), acetate, and chloroform fractions, as well as the catuabine B, increased in the number of cells in early apoptosis, from 53.0 to 74.8%. An analysis of the potential of the mitochondrial membrane by incorporation of JC-1 showed that most cells during incubation of the acetate: methanol (95:5) and acetate fractions (63.85 and 59.2%) were stained, suggesting the involvement of an intrinsic pathway of apoptosis.

Cytotoxic effect and apoptosis induction by Bothrops leucurus venom lectin on tumor cell lines.

Neoplastic transformation is the abnormal proliferation of cells. These transformations are often related to changes in cell surface glycoconjugates which can be detected by lectins. We evaluated the anti-tumor potential of BlL, a galactoside-binding lectin isolated from Bothrops leucurus venom as well as its cytotoxicity and hemolysis activity. The phosphatidylserine externalization and mitochondrial membrane potential were also determined. BlL exhibited cytotoxic activity against all tumor cell lines tested by induced phosphatidylserine externalization and mitochondrial depolarization, indicating cell death by apoptosis.

A cell junction pathology of neural stem cells leads to abnormal neurogenesis and hydrocephalus.

Most cells of the developing mammalian brain derive from the ventricular (VZ) and the subventricular (SVZ) zones. The VZ is formed by the multipotent radial glia/neural stem cells (NSCs) while the SVZ harbors the rapidly proliferative neural precursor cells (NPCs). Evidence from human and animal models indicates that the common history of hydrocephalus and brain maldevelopment starts early in embryonic life with disruption of the VZ and SVZ. We propose that a "cell junction pathology" involving adherent and gap junctions is a final common outcome of a wide range of gene mutations resulting in proteins abnormally expressed by the VZ cells undergoing disruption. Disruption of the VZ during fetal development implies the loss of NSCs whereas VZ disruption during the perinatal period implies the loss of ependyma. The process of disruption occurs in specific regions of the ventricular system and at specific stages of brain development. This explains why only certain brain structures have an abnormal development, which in turn results in a specific neurological impairment of the newborn. Disruption of the VZ of the Sylvian aqueduct (SA) leads to aqueductal stenosis and hydrocephalus, while disruption of the VZ of telencephalon impairs neurogenesis. We are currently investigating whether grafting of NSCs/neurospheres from normal rats into the CSF of hydrocephalic mutants helps to diminish/repair the outcomes of VZ disruption.

A cell junction pathology of neural stem cells leads to abnormal neurogenesis and hydrocephalus

Most cells of the developing mammalian brain derive from the ventricular (VZ) and the subventricular (SVZ) zones. The VZ is formed by the multipotent radial glia/neural stem cells (NSCs) while the SVZ harbors the rapidly proliferative neural precursor cells (NPCs). Evidence from human and animal models indicates that the common history of hydrocephalus and brain maldevelopment starts early in embryonic life with disruption of the VZ and SVZ. We propose that a "cell junction pathology" involving adherent and gap junctions is a final common outcome of a wide range of gene mutations resulting in proteins abnormally expressed by the VZ cells undergoing disruption. Disruption of the VZ during fetal development implies the loss of NSCs whereas VZ disruption during the perinatal period implies the loss of ependyma. The process of disruption occurs in specific regions of the ventricular system and at specific stages of brain development. This explains why only certain brain structures have an abnormal development, which in turn results in a specific neurological impairment of the newborn. Disruption of the VZ of the Sylvian aqueduct (SA) leads to aqueductal stenosis and hydrocephalus, while disruption of the VZ of telencephalon impairs neurogenesis. We are currently investigating whether grafting of NSCs/neurospheres from normal rats into the CSF of hydrocephalic mutants helps to diminish/repair the outcomes of VZ disruption.

Prevalência e fatores de risco associados à infecção por Chlamydophila abortus em granjas suinícolas tecnificadas no estado de Alagoas, Brasil / Prevalence and risk factors associated with infection by Chlamydophila abortus in commercial pig farms in the state of Alagoas, Brazil

Objetivou-se com este estudo calcular a prevalência e identificar os fatores de risco associados à infecção por Chlamydophila abortus em suínos criados em granjas tecnificadas no Estado de Alagoas, Brasil. Para compor a amostra do estudo foram utilizados 342 suínos, sendo 312 matrizes e 30 varrões oriundos de sete granjas de ciclo completo e distribuídas em cinco municípios do Estado de Alagoas. O diagnóstico sorológico da infecção por C. abortus foi realizado através da microtécnica de Fixação do Complemento (RFC). A análise dos fatores de risco foi realizada por meio da aplicação de questionários investigativos, constituídos por perguntas objetivas referentes ao criador, às características gerais da propriedade, ao manejo produtivo, reprodutivo e sanitário. Observou-se prevalência de 10,5% (36/342) de suínos soropositivos para a infecção por C. abortus, com 85,8% das granjas analisadas com animais positivos. As variáveis que demonstraram associação significativa foram: utilização de bebedouros comuns para jovens e adultos (p=0,024;OR=10,83; IC=1,36–86,03) e método de cobertura de monta natural associada à inseminação artificial (p=0,05; OR=7,62; IC=1,00-58,31). Relata-se a primeira ocorrência de anticorpos anti-C. abortus em suínos no Brasil. Fatores como a introdução de reprodutores nos plantéis e a forma de fornecimento de água foram evidenciados como facilitadores da infecção das matrizes neste estudo. Dessa forma, medidas de controle da infecção devem ser enfocadas nesse aspecto para evitar a disseminação do agente nas granjas suinícolas e em outros plantéis da região.

The objective of this study was to determine the prevalence and identify risk factors associated with Chlamydophila abortus infection in commercial swine farms on the state of Alagoas, Brazil. To compose the study sample 342 pigs were used, with 312 sows and 30 boars from seven swine farms and distributed in five districts of the Alagoas. The serological diagnosis of infection by C. abortus was performed by fixation of complement microtechnique (RFC). The analysis of risk factors were performed by the application of research questionnaires, consisting of objective questions relating to the designer, the general characteristics of the property, the production, reproductive and health management. I was observed a prevalence of 10.5% (36/342) of pigs seropositive for infection by C. abortus with 85.8% of farms with positive animals analyzed. The variables that showed significant association were use of common drinker for young and adults pigs (p=0.024, OR=10.83, CI=1.36-86.03) and associated the natural mount with artificial insemination (p=0.05, OR=7.62, CI=1.00-58.31). This work reports the first occurrence of anti-C. abortusin pigs in Brazil. Factors as the introduction of boars in herds and the form of water supply were seen as facilitators of infection on sows in this study. Thus measures of infection control should be focused on this aspect to prevent the spread of the agent in pig farms and other herds in the region.

Biodisponibilidad y bioutilización de mezclas de aminoácidos vs. proteínas intactas / Bioavailability and bioutilization of mixtures of amino acids vs. intact proteins

La absorción y utilización del nitrógeno de la dieta fueron estudiadas en ratas en el crecimiento cuando este elemento se suministra en forma de proteina intacta o de mezclas equivalentas de aminoácidos. Las mezclas de aminoácidos ingeridas son absorbidas más eficientemente y ocasionan en el organismo similares valores de deposición proteica, los cuales se logran a expensas de un incrementado consumo de energia y de una redución del compartimiento graso del organismo. La reducción de la ingestión de la dieta y el elevado consumo energético que genera la bioutilización de los aminoácidos aislados no mejora con el suplemento de minerales y oligoelementos. La sustitución de los aminoácidos no esenciales de la dieta por ácido glutámico como única fuente de nitrógeno no esencial, reduce considerablemente la bioutilización del nitrógeno de la dieta

Biodisponibilidad y bioutilización de mezclas de aminoácidos vs. proteínas intactas / Bioavailability and bioutilization of mixtures of amino acids vs. intact proteins

La absorción y utilización del nitrógeno de la dieta fueron estudiadas en ratas en el crecimiento cuando este elemento se suministra en forma de proteina intacta o de mezclas equivalentas de aminoácidos. Las mezclas de aminoácidos ingeridas son absorbidas más eficientemente y ocasionan en el organismo similares valores de deposición proteica, los cuales se logran a expensas de un incrementado consumo de energia y de una redución del compartimiento graso del organismo. La reducción de la ingestión de la dieta y el elevado consumo energético que genera la bioutilización de los aminoácidos aislados no mejora con el suplemento de minerales y oligoelementos. La sustitución de los aminoácidos no esenciales de la dieta por ácido glutámico como única fuente de nitrógeno no esencial, reduce considerablemente la bioutilización del nitrógeno de la dieta