Neutrofilia e hipoalbuminemia em sangue periférico de cães naturalmente infectados por Leishmania infantum / Neutrophilia and hypoalbuminemia in peripheral blood of dogs naturally infected by Leishmania infantum

O objetivo deste estudo foi caracterizar o perfil leucocitário e proteico de cães, naturalmente infectados por Leishmania infantum. Para tanto, foram realizados exame parasitológico de punção de medula e PCR em amostras de pele para identificação de Leishmania spp. e avaliação de sinais clínicos e dermatológicos, para categorizar e distribuir cães em três grupos: grupo controle (GC); Grupo positivo assintomático (GA) e grupo positivo sintomático (GS). Amostras De sangue foram coletadas para avaliação dos parâmetros hematológicos e bioquímicos. As principais manifestações clínicas observadas no GS foram alopecia, onicogrifose, ceratoconjuntivite e dermatites eritematosas, esfoliativas e ulcerativas. Os animais com a forma visceral (LVC) apresentaram alterações hematológicas significativas, compatíveis com descrições da literatura, com leucocitose significativa (p<0,05) do GS e do GA quando comparados ao GC, além de anemia no GS e GA, em relação ao GC. Também foram observadas hiperproteinemia, hipergamablobulinemia e hipoalbuminemia significativas em GS, quando comparadas aos demais grupos (p<0,05). A contagem diferencial leucocitária apontou para uma neutrofilia e linfopenia significativas do GS, quando comparadas aos demais grupos (p<0,05), Estando este resultado confirmado pela razão neutrófilo: linfócito aumentada (p<0,05). Ressalta-se um aumento de neutrófilos segmentados maduros no Leucograma, mas sem alterações significativas nos neutrófilos imaturos. Animais naturalmente infectados por L. infantum apresentaram leucocitose com neutrofilia e hipoalbuminemia características de alterações sistêmicas associadas a um processo inflamatório. Contudo, faz-se necessária uma investigação mais aprofundada, com um maior número de animais e com acompanhamento da LVC para avaliar a participação dos neutrófilos imaturos neste processo.(AU)

The Objective of this study was to evaluate the leukocyte and protein profile of dogs naturally infected by Leishmania Infantum. For that, a bone marrow parasitological test and PCR were performed to identify Leishmania spp. Evaluation of clinical and dermatological signs was used to categorize and distribute dogs in three groups: control group (CG); asymptomatic group (AG) and symptomatic group (SG). Blood samples were collected for evaluation of hematological and biochemical parameters.The Main clinical manifestations observed in SG were alopecia, onychogryphosis, keratoconjunctivitis and dermatitis. Leukocytosis and anemia were observed in SG and AG (p<0.05) when compared to CG. In SG were detected hyperproteinemia with hyperglobulinemia and hypoalbuminemia when compared to the other groups (p<0.05). The differential leukocytes count indicated neutrophilia and lymphopenia in SG when compared to AG and CG (p<0.05), and this result was confirmed bya neutrophil:lymphocyte ratio which was increased (p<0.05). We Highlight an increase of mature segmented neutrophils in GS, but there is no change in immature neutrophils. Thus, animals naturally infected by L. infantum presented leukocytosis with neutrophilia and hypoalbuminemia characteristic of systemic changes associated to inflammatory process. However, a more in-depth investigation with a larger number of animals and earlier follow-Up of CanL Animals is necessary to evaluate the participation of immature neutrophils in this process.(AU)

Neutrofilia e hipoalbuminemia em sangue periférico de cães naturalmente infectados por Leishmania infantum / Neutrophilia and hypoalbuminemia in peripheral blood of dogs naturally infected by Leishmania infantum

O objetivo deste estudo foi caracterizar o perfil leucocitário e proteico de cães, naturalmente infectados por Leishmania infantum. Para tanto, foram realizados exame parasitológico de punção de medula e PCR em amostras de pele para identificação de Leishmania spp. e avaliação de sinais clínicos e dermatológicos, para categorizar e distribuir cães em três grupos: grupo controle (GC); Grupo positivo assintomático (GA) e grupo positivo sintomático (GS). Amostras De sangue foram coletadas para avaliação dos parâmetros hematológicos e bioquímicos. As principais manifestações clínicas observadas no GS foram alopecia, onicogrifose, ceratoconjuntivite e dermatites eritematosas, esfoliativas e ulcerativas. Os animais com a forma visceral (LVC) apresentaram alterações hematológicas significativas, compatíveis com descrições da literatura, com leucocitose significativa (p<0,05) do GS e do GA quando comparados ao GC, além de anemia no GS e GA, em relação ao GC. Também foram observadas hiperproteinemia, hipergamablobulinemia e hipoalbuminemia significativas em GS, quando comparadas aos demais grupos (p<0,05). A contagem diferencial leucocitária apontou para uma neutrofilia e linfopenia significativas do GS, quando comparadas aos demais grupos (p<0,05), Estando este resultado confirmado pela razão neutrófilo: linfócito aumentada (p<0,05). Ressalta-se um aumento de neutrófilos segmentados maduros no Leucograma, mas sem alterações significativas nos neutrófilos imaturos. Animais naturalmente infectados por L. infantum apresentaram leucocitose com neutrofilia e hipoalbuminemia características de alterações sistêmicas associadas a um processo inflamatório. Contudo, faz-se necessária uma investigação mais aprofundada, com um maior número de animais e com acompanhamento da LVC para avaliar a participação dos neutrófilos imaturos neste processo.

The Objective of this study was to evaluate the leukocyte and protein profile of dogs naturally infected by Leishmania Infantum. For that, a bone marrow parasitological test and PCR were performed to identify Leishmania spp. Evaluation of clinical and dermatological signs was used to categorize and distribute dogs in three groups: control group (CG); asymptomatic group (AG) and symptomatic group (SG). Blood samples were collected for evaluation of hematological and biochemical parameters.The Main clinical manifestations observed in SG were alopecia, onychogryphosis, keratoconjunctivitis and dermatitis. Leukocytosis and anemia were observed in SG and AG (p<0.05) when compared to CG. In SG were detected hyperproteinemia with hyperglobulinemia and hypoalbuminemia when compared to the other groups (p<0.05). The differential leukocytes count indicated neutrophilia and lymphopenia in SG when compared to AG and CG (p<0.05), and this result was confirmed bya neutrophil:lymphocyte ratio which was increased (p<0.05). We Highlight an increase of mature segmented neutrophils in GS, but there is no change in immature neutrophils. Thus, animals naturally infected by L. infantum presented leukocytosis with neutrophilia and hypoalbuminemia characteristic of systemic changes associated to inflammatory process. However, a more in-depth investigation with a larger number of animals and earlier follow-Up of CanL Animals is necessary to evaluate the participation of immature neutrophils in this process.

Avanços no isolamento e sistemas de cultivo de folículos pré-antrais / Advances in isolation and culture systems of preantral follicles

Os folículos pré-antrais presentes no córtex ovariano representam cerca de 95% de toda a população folicular e podem ser crescidos e maturados in vitro, visando a obtenção posterior de oócitos maturos e competentes, os quais poderão ser utilizados nas diversas biotécnicas da reprodução aplicadas aos animais domésticos. Os estudos relacionados com a foliculogênese podem ser feitos in vivo, através da análise de expressão de genes ou proteínas que estão presentes no ovário, e ainda in vitro com a utilização de diferentes sistemas, onde os folículos podem ser cultivados in situ ou isolados. Deste modo, neste trabalho foram abordados os diferentes métodos de isolamento e sistemas de cultivo in vitro empregados para a manutenção da integridade estrutural e desenvolvimento de folículos pré-antrais.

The preantral follicles enclosed in ovarian cortex represent about 95% of the entire follicular population and can be grown and matured in vitro in order to obtain mature and competent oocytes, which may be used in various reproductive biotechnologies applied to domestic animals. Studies related to folliculogenesis can be performed in vivo, by analyzing the expression of genes or proteins that are present in the ovary, and also in vitro with the use of different systems, in which the follicles can be grown in situ or isolated. Thus, this study shows the different methods of isolation and in vitro systems employed for maintaining the structural integrity and development of preantral follicles.

Ativina intraovariana: uma atualização sobre estrutura, regulação e função / Intraovarian activin: an update on structure, regulation and function

Background: Important advances have been made recently that clarify our understanding of the structural basis, signaling and regulation, as well as the biological role of activin in ovaries. During folliculogenesis various growth factors are produced locally in the mammalian ovary. Among these factors, activin has been a focal point in research as it has emerged as a crucial substance capable of inducing follicular development. The important actions indicate that activin has many relevant homeostatic functions in the reproduction of several species. Therefore, this review discusses the ligand protein structure, activin receptors, mechanisms of action and regulation, as well as the importance of activin on in vitro culture of preantral follicles. Review: Activin belongs to the transforming growth factor β (TGF - β) super family. It is a homodimer or heterodimer of two similar but distinct subunits (βA and βB). The dimerisation of activin subunits gives rise to three forms of activin, which are classified as, activin A (βA - βA), activin B (βB - βB) and activin AB (βA - βB). The biological activity of activin occurs through its connection with two types of cell surface receptors designated type I and type II. These receptors are represented by two isoforms, activin receptor types IA (ActR - IA), IB (ActR - IB), IIA (ActR - IIA) and IIB (ActR - IIB). Activin receptors are transmembrane proteins, composed of a ligand-binding extracellular domain, a transmembrane domain and a cytoplasmic domain with serine/threonine kinase activity. The transient activation of the receptor induces phosphorylation of protein mediators called Smads. Activation of Smad 2/3 by phosphorylation causes trimerization and hetero-oligomerization with the common Smad, Smad 4. This complex translocates to the nucleus to activating and regulating transcription of target genes. Members of another class of Smads act as negative regulators of the signal transduction pathway. Inhibitory Smad 7 can bind to type I receptors, preventing receptor–Smad 2/3 association, or by competitively binding of Smad 4, which blocks Smad intracellular translocation. In addition, within the extracellular environment, binding proteins such as follistatin and inhibin can modulate the biological activity of activin. In the ovaries of mammals specifically, activin participates in several cellular events, including cellular proliferation, differentiation, and survival, as well as assisting steroidal hormones during follicular development. Activin has been localized in the oocytes and granulosa cells of rodent, porcine, caprine and bovine follicles. Activin is also within the granulosa cells of human follicles and in the thecal layers of porcine and human. In addition, activin stimulates follicle growth in-vitro, is used in pre-antral ovine and caprine follicles and enhances growth and survival of human pre-antral follicles in vitro. Conclusion: Activin is controlled by competitive substances and a dynamic interaction between the various regulatory proteins responsible for coordinating several signaling pathways. The balance between the actions of these proteins is critical for regulation of gene expression in different structures, including pre-antral follicles. However, the nature of physiological effects of activin in the ovary is still equivocal and awaits clarification.(AU)

Ativina intraovariana: uma atualização sobre estrutura, regulação e função / Intraovarian activin: an update on structure, regulation and function

Background: Important advances have been made recently that clarify our understanding of the structural basis, signaling and regulation, as well as the biological role of activin in ovaries. During folliculogenesis various growth factors are produced locally in the mammalian ovary. Among these factors, activin has been a focal point in research as it has emerged as a crucial substance capable of inducing follicular development. The important actions indicate that activin has many relevant homeostatic functions in the reproduction of several species. Therefore, this review discusses the ligand protein structure, activin receptors, mechanisms of action and regulation, as well as the importance of activin on in vitro culture of preantral follicles. Review: Activin belongs to the transforming growth factor β (TGF - β) super family. It is a homodimer or heterodimer of two similar but distinct subunits (βA and βB). The dimerisation of activin subunits gives rise to three forms of activin, which are classified as, activin A (βA - βA), activin B (βB - βB) and activin AB (βA - βB). The biological activity of activin occurs through its connection with two types of cell surface receptors designated type I and type II. These receptors are represented by two isoforms, activin receptor types IA (ActR - IA), IB (ActR - IB), IIA (ActR - IIA) and IIB (ActR - IIB). Activin receptors are transmembrane proteins, composed of a ligand-binding extracellular domain, a transmembrane domain and a cytoplasmic domain with serine/threonine kinase activity. The transient activation of the receptor induces phosphorylation of protein mediators called Smads. Activation of Smad 2/3 by phosphorylation causes trimerization and hetero-oligomerization with the common Smad, Smad 4. This complex translocates to the nucleus to activating and regulating transcription of target genes. Members of another class of Smads act as negative regulators of the signal transduction pathway. Inhibitory Smad 7 can bind to type I receptors, preventing receptor–Smad 2/3 association, or by competitively binding of Smad 4, which blocks Smad intracellular translocation. In addition, within the extracellular environment, binding proteins such as follistatin and inhibin can modulate the biological activity of activin. In the ovaries of mammals specifically, activin participates in several cellular events, including cellular proliferation, differentiation, and survival, as well as assisting steroidal hormones during follicular development. Activin has been localized in the oocytes and granulosa cells of rodent, porcine, caprine and bovine follicles. Activin is also within the granulosa cells of human follicles and in the thecal layers of porcine and human. In addition, activin stimulates follicle growth in-vitro, is used in pre-antral ovine and caprine follicles and enhances growth and survival of human pre-antral follicles in vitro. Conclusion: Activin is controlled by competitive substances and a dynamic interaction between the various regulatory proteins responsible for coordinating several signaling pathways. The balance between the actions of these proteins is critical for regulation of gene expression in different structures, including pre-antral follicles. However, the nature of physiological effects of activin in the ovary is still equivocal and awaits clarification.