Effect of cold stress on physicochemical characteristics and biological activity of equine chorionic gonadotropin

The purpose of this study was to evaluate iffreezing-thawing and cooling processes affect thestructural properties and biological activity ofcommercial equine chorionic gonadotropin (eCG). First,the structure profile of diluted eCG underwent none,one or three cycles of freezing-thawing was analysed byreverse phase high-performance liquid chromatography(RP-HPLC). In a second experiment, groups ofprepuberal rats were treated with sterile water forinjection USP or eCG that underwent none, one or threecycles of freezing-thawing to assess the increase ofovarian weigh. Finally, groups of prepubertal gilts weretreated with diluted eCG immediately afterreconstitution (T1), after refrigeration for six months(T2) and after freezing and subsequently thawing for one(T3) or three (T4) cycles. The control group (T5) receivedsterile water for injection USP without eCG. Ovulationwas induced with human chorionic gonadotropin (hCG),administered 72 h after the eCG. Gilts were slaughteredfive days after the hCG injection and ovaries wererecovered and analysed for the presence of corporalutea. Data were analysed by ANOVA and Fishersexact tests. In the analyses by RP-HPLC, the retentiontimes of cold stressed eCG were similar to unstressedcontrol. The mean ovarian weight of rats treated with coldstressed and unstressed eCG was statistically higher thanwater control (P < 0.05). Lastly, significantly more giltsovulated in groups T1, T2, T3 and T4 than in thecontrol T5 (P < 0.05). It was concluded that freezingthawing,as well as cooling over a period of up to sixmonths, did not significantly change the structuralproperties or biological activity of eCG.(AU)

Effect of cold stress on physicochemical characteristics and biological activity of equine chorionic gonadotropin

The purpose of this study was to evaluate iffreezing-thawing and cooling processes affect thestructural properties and biological activity ofcommercial equine chorionic gonadotropin (eCG). First,the structure profile of diluted eCG underwent none,one or three cycles of freezing-thawing was analysed byreverse phase high-performance liquid chromatography(RP-HPLC). In a second experiment, groups ofprepuberal rats were treated with sterile water forinjection USP or eCG that underwent none, one or threecycles of freezing-thawing to assess the increase ofovarian weigh. Finally, groups of prepubertal gilts weretreated with diluted eCG immediately afterreconstitution (T1), after refrigeration for six months(T2) and after freezing and subsequently thawing for one(T3) or three (T4) cycles. The control group (T5) receivedsterile water for injection USP without eCG. Ovulationwas induced with human chorionic gonadotropin (hCG),administered 72 h after the eCG. Gilts were slaughteredfive days after the hCG injection and ovaries wererecovered and analysed for the presence of corporalutea. Data were analysed by ANOVA and Fisher’sexact tests. In the analyses by RP-HPLC, the retentiontimes of cold stressed eCG were similar to unstressedcontrol. The mean ovarian weight of rats treated with coldstressed and unstressed eCG was statistically higher thanwater control (P < 0.05). Lastly, significantly more giltsovulated in groups T1, T2, T3 and T4 than in thecontrol T5 (P < 0.05). It was concluded that freezingthawing,as well as cooling over a period of up to sixmonths, did not significantly change the structuralproperties or biological activity of eCG.

Indução da ovulação e características ovarianas de marrãs pré-púberes tratadas com gonadotrofina coriônica humana ou kisspeptina / Induction of ovulation and ovarian characteristics of prepubertal gilts treated with human chorionicgonadotropin or kisspeptin

Devido à semelhança na estrutura e função com o hormônio luteinizante (LH), a gonadotrofina coriônica humana (hCG) tem sido convencionalmente o principal hormônio utilizado para induzir o estro e a ovulação nas fêmeas suínas. Recentemente, foi demonstrado que o neuropeptídio kisspeptina é o precursor da descarga de LH em diferentes espécies de mamíferos, incluindo a suína. Dessa forma, o presente estudo objetivou determinar se uma das formas dessa substância (kisspeptina-10) associada à gonadotrofina coriônica equina (eCG) pode ser um método eficiente para induzir a ovulação de marrãs pré-púberes. Trinta marrãs de 5 meses de idade, com peso médio de 78,0 ± 8,4 kg, foram distribuídas aleatoriamente em cinco grupos de seis animais cada. Os grupos 1, 2 e 3 receberam 750 UI de eCG, enquanto que os grupos 4 e 5 receberam solução fisiológica. Setenta e duas horas após, o grupo 2 recebeu 380 nmol de kisspeptina-10, os grupos 3 e 4 receberam 500 UI de hCG e os grupo 1 e 5 receberam solução fisiológica. As injeções foram aplicadas em dose única, utilizando a via intramuscular. Os animais foram abatidos sete dias após e a ovulação foi constatada pela presença de corpos lúteos nos ovários recuperados. Os dados foram analisados pelo teste de Kruscal-Wallis e teste exato de Fisher. O número de fêmeas que ovulou foi semelhante nos grupos 2 e 3. Essa resposta foi superior ao grupo 5 (P<0,01) e aos grupos 1 e 4 (P=0,08). O peso e o tamanho dos ovários não foram influenciados pelos tratamentos, enquanto que o número de ovulações foi maior para os grupos 2 e 3 (P<0,01). Esses resultados sugerem que a kisspeptina-10 tem atividade biológica comparável à hCG para induzir a ovulação de marrãs pré-púberes...

In view of its similar structure and function to luteinizing hormone (LH), human chorionic gonadotropin (hCG) has conventionally been used as the main hormone to induce estrus and ovulation in sows. Recently, it has been demonstrated that the neuropeptide kisspeptin is associated with pituitary LH release in different mammalian species, including swine. Thus, the aim of this study was to determine whether kisspeptin-10 combined with equine chorionic gonadotropin (eCG) could be an efficient method to induce ovulation in prepubertal gilts. Thirty 5-month-old gilts weighing 78.0 ± 8.4 kg were randomly divided into five groups of six animals each. Groups 1, 2 and 3 received 750 IU eCG, while groups 4 and 5 received saline solution. After 72 hours, the animals received 380 nmol kisspeptin-10 (group 2), 500 IU hCG (groups 3 and 4), or saline (groups 1 and 5). The treatments were applied by a single intramuscular injection. The animals were slaughtered 7 days later and ovulation was confirmed by the presence of corpora lutea in the recovered ovaries. Data were analyzed by the Kruskal-Wallis and Fisher exact tests. The number of gilts that ovulated was similar in groups 2 and 3. This number was higher in group 5 (P<0.01) and in groups 1 and 4 (P=0.08). The weight or size of the ovaries was not influenced by the treatments, although the number of ovulations was higher in groups 2 and 3 (P<0.01). These results suggest that kisspeptin-10 and hCG have similar biological activity to induce ovulation in prepubertal gilts...

Indução da ovulação e características ovarianas de marrãs pré-púberes tratadas com gonadotrofina coriônica humana ou kisspeptina / Induction of ovulation and ovarian characteristics of prepubertal gilts treated with human chorionicgonadotropin or kisspeptin

Devido à semelhança na estrutura e função com o hormônio luteinizante (LH), a gonadotrofina coriônica humana (hCG) tem sido convencionalmente o principal hormônio utilizado para induzir o estro e a ovulação nas fêmeas suínas. Recentemente, foi demonstrado que o neuropeptídio kisspeptina é o precursor da descarga de LH em diferentes espécies de mamíferos, incluindo a suína. Dessa forma, o presente estudo objetivou determinar se uma das formas dessa substância (kisspeptina-10) associada à gonadotrofina coriônica equina (eCG) pode ser um método eficiente para induzir a ovulação de marrãs pré-púberes. Trinta marrãs de 5 meses de idade, com peso médio de 78,0 ± 8,4 kg, foram distribuídas aleatoriamente em cinco grupos de seis animais cada. Os grupos 1, 2 e 3 receberam 750 UI de eCG, enquanto que os grupos 4 e 5 receberam solução fisiológica. Setenta e duas horas após, o grupo 2 recebeu 380 nmol de kisspeptina-10, os grupos 3 e 4 receberam 500 UI de hCG e os grupo 1 e 5 receberam solução fisiológica. As injeções foram aplicadas em dose única, utilizando a via intramuscular. Os animais foram abatidos sete dias após e a ovulação foi constatada pela presença de corpos lúteos nos ovários recuperados. Os dados foram analisados pelo teste de Kruscal-Wallis e teste exato de Fisher. O número de fêmeas que ovulou foi semelhante nos grupos 2 e 3. Essa resposta foi superior ao grupo 5 (P<0,01) e aos grupos 1 e 4 (P=0,08). O peso e o tamanho dos ovários não foram influenciados pelos tratamentos, enquanto que o número de ovulações foi maior para os grupos 2 e 3 (P<0,01). Esses resultados sugerem que a kisspeptina-10 tem atividade biológica comparável à hCG para induzir a ovulação de marrãs pré-púberes...(AU)

In view of its similar structure and function to luteinizing hormone (LH), human chorionic gonadotropin (hCG) has conventionally been used as the main hormone to induce estrus and ovulation in sows. Recently, it has been demonstrated that the neuropeptide kisspeptin is associated with pituitary LH release in different mammalian species, including swine. Thus, the aim of this study was to determine whether kisspeptin-10 combined with equine chorionic gonadotropin (eCG) could be an efficient method to induce ovulation in prepubertal gilts. Thirty 5-month-old gilts weighing 78.0 ± 8.4 kg were randomly divided into five groups of six animals each. Groups 1, 2 and 3 received 750 IU eCG, while groups 4 and 5 received saline solution. After 72 hours, the animals received 380 nmol kisspeptin-10 (group 2), 500 IU hCG (groups 3 and 4), or saline (groups 1 and 5). The treatments were applied by a single intramuscular injection. The animals were slaughtered 7 days later and ovulation was confirmed by the presence of corpora lutea in the recovered ovaries. Data were analyzed by the Kruskal-Wallis and Fisher exact tests. The number of gilts that ovulated was similar in groups 2 and 3. This number was higher in group 5 (P<0.01) and in groups 1 and 4 (P=0.08). The weight or size of the ovaries was not influenced by the treatments, although the number of ovulations was higher in groups 2 and 3 (P<0.01). These results suggest that kisspeptin-10 and hCG have similar biological activity to induce ovulation in prepubertal gilts...(AU)

Eficiência da inseminação artificial em tempo fixo utilizando dispositivo de progesterona associado com GnRH ou benzoato de estradiol em novilhas da raça Nelore / Efficiency of fixed-time artificial insemination using a progesterone device combined with GnRH or estradiol benzoate in Nellore heifers

O emprego de estrógenos nos protocolos hormonais para programar a inseminação artificial dos bovinos é o método mais barato e eficiente atualmente disponível. Entretanto, a tendência de proibir o uso de estrógenos para esse propósito torna necessária a procura por alternativas que substituam os estrógenos sem prejudicar o desempenho reprodutivo dos animais. O objetivo deste estudo foi avaliar as taxas de concepção de novilhas de corte Bos indicus tratadas com dispositivo de progesterona (P4) associado ao GnRH ou a um éster de estradiol. Novilhas púberes da raça Nelore (n = 100) foram tratadas no dia 0 com um dispositivo intravaginal contendo 1 g de P4 e distribuídas aleatoriamente em dois grupos. O grupo GnRH (n = 49) recebeu uma injeção im de 100 µg de GnRH, enquanto que o grupo E2 (n = 51) recebeu 2 mg de benzoato de estradiol (BE). O dispositivo de P4 foi removido após 5 (grupo GnRH) ou 8 (grupo E2) dias, seguido de uma injeção de 125 µg do análogo de PGF2α, cloprostenol. Nesse momento, o grupo E2 recebeu, adicionalmente, 300 UI de eCG. Vinte e quatro horas depois, o grupo GnRH recebeu uma segunda injeção de 125 µg de cloprostenol, enquanto que o grupo E2 recebeu 1 mg de BE. As novilhas foram inseminadas 72 (grupo GnRH) ou 54 (grupo E2) horas após a retirada do dispositivo de P4 e, no momento da inseminação, o grupo GnRH recebeu adicionalmente uma injeção de 100 µg de GnRH. O cio foi monitorado no período da injeção de cloprostenol até o momento da inseminação artificial (IA) e o diagnóstico de prenhez foi realizado 40 dias após a IA, utilizando ultrassonografia transretal. Os dados foram analisados pelo teste exato de Ficher...

The use of estrogens in artificial insemination protocols for cattle is the least expensive and most efficient method currently available. However, the trend to prohibit the use of estrogens for this purpose has made it necessary to find alternatives that replace estrogens without compromising the reproductive performance of the animals. The objective of this study was to evaluate conception rates in Bos indicus beef heifers treated with a progesterone device (P4) combined with GnRH or an estradiol ester. On day 0, pubertal Nellore heifers (n = 100) received an intravaginal device containing 1 g P4 and were randomly divided into two groups. The GnRH group (n = 49) received an intramuscular injection of 100 µg GnRH, while the E2 group (n = 51) received 2 mg estradiol benzoate (EB). The P4 device was removed after 5 (GnRH group) or 8 days (E2 group), followed by an injection of 125 µg of the PGF2α, analog cloprostenol. On that occasion, the E2 group received an additional injection of 300 IU eCG. Twenty-four hours later, the GnRH group received a second injection of 125 µg cloprostenol, while the E2 group received 1 mg EB. The heifers were inseminated 72 (GnRH group) or 54 hours (E2 group) after removal of the P4 device. At the time of insemination, the GnRH group received additionally an injection of 100 µg GnRH. Estrus was monitored during the period of cloprostenol injection until the time of artificial insemination and pregnancy was diagnosed 40 days after insemination by transrectal ultrasonography...

Eficiência da inseminação artificial em tempo fixo utilizando dispositivo de progesterona associado com GnRH ou benzoato de estradiol em novilhas da raça Nelore / Efficiency of fixed-time artificial insemination using a progesterone device combined with GnRH or estradiol benzoate in Nellore heifers

O emprego de estrógenos nos protocolos hormonais para programar a inseminação artificial dos bovinos é o método mais barato e eficiente atualmente disponível. Entretanto, a tendência de proibir o uso de estrógenos para esse propósito torna necessária a procura por alternativas que substituam os estrógenos sem prejudicar o desempenho reprodutivo dos animais. O objetivo deste estudo foi avaliar as taxas de concepção de novilhas de corte Bos indicus tratadas com dispositivo de progesterona (P4) associado ao GnRH ou a um éster de estradiol. Novilhas púberes da raça Nelore (n = 100) foram tratadas no dia 0 com um dispositivo intravaginal contendo 1 g de P4 e distribuídas aleatoriamente em dois grupos. O grupo GnRH (n = 49) recebeu uma injeção im de 100 µg de GnRH, enquanto que o grupo E2 (n = 51) recebeu 2 mg de benzoato de estradiol (BE). O dispositivo de P4 foi removido após 5 (grupo GnRH) ou 8 (grupo E2) dias, seguido de uma injeção de 125 µg do análogo de PGF2α, cloprostenol. Nesse momento, o grupo E2 recebeu, adicionalmente, 300 UI de eCG. Vinte e quatro horas depois, o grupo GnRH recebeu uma segunda injeção de 125 µg de cloprostenol, enquanto que o grupo E2 recebeu 1 mg de BE. As novilhas foram inseminadas 72 (grupo GnRH) ou 54 (grupo E2) horas após a retirada do dispositivo de P4 e, no momento da inseminação, o grupo GnRH recebeu adicionalmente uma injeção de 100 µg de GnRH. O cio foi monitorado no período da injeção de cloprostenol até o momento da inseminação artificial (IA) e o diagnóstico de prenhez foi realizado 40 dias após a IA, utilizando ultrassonografia transretal. Os dados foram analisados pelo teste exato de Ficher...(AU)

The use of estrogens in artificial insemination protocols for cattle is the least expensive and most efficient method currently available. However, the trend to prohibit the use of estrogens for this purpose has made it necessary to find alternatives that replace estrogens without compromising the reproductive performance of the animals. The objective of this study was to evaluate conception rates in Bos indicus beef heifers treated with a progesterone device (P4) combined with GnRH or an estradiol ester. On day 0, pubertal Nellore heifers (n = 100) received an intravaginal device containing 1 g P4 and were randomly divided into two groups. The GnRH group (n = 49) received an intramuscular injection of 100 µg GnRH, while the E2 group (n = 51) received 2 mg estradiol benzoate (EB). The P4 device was removed after 5 (GnRH group) or 8 days (E2 group), followed by an injection of 125 µg of the PGF2α, analog cloprostenol. On that occasion, the E2 group received an additional injection of 300 IU eCG. Twenty-four hours later, the GnRH group received a second injection of 125 µg cloprostenol, while the E2 group received 1 mg EB. The heifers were inseminated 72 (GnRH group) or 54 hours (E2 group) after removal of the P4 device. At the time of insemination, the GnRH group received additionally an injection of 100 µg GnRH. Estrus was monitored during the period of cloprostenol injection until the time of artificial insemination and pregnancy was diagnosed 40 days after insemination by transrectal ultrasonography...(AU)

Genetic diversity of polysporic isolates of Moniliophthora perniciosa (Tricholomataceae).

The causal agent of witches' broom disease, Moniliophthora perniciosa is a hemibiotrophic and endemic fungus of the Amazon basin and the most important cocoa disease in Brazil. The purpose of this study was to analyze the genetic diversity of polysporic isolates of M. perniciosa to evaluate the adaptation of the pathogen from different Brazilian regions and its association with different hosts. Polysporic isolates obtained previously in potato dextrose agar cultures of M. perniciosa from different Brazilian states and different hosts (Theobroma cacao, Solanum cernuum, S. paniculatum, S. lycocarpum, Solanum sp, and others) were analyzed by somatic compatibility grouping where the mycelium interactions were distinguished after 4-8 weeks of confrontation between the different isolates of M. perniciosa based on the precipitation line in the transition zone and by protein electrophoresis through SDS-PAGE. The diversity of polysporic isolates of M. perniciosa was grouped according to geographic proximity and respective hosts. The great genetic diversity of M. perniciosa strains from different Brazilian states and hosts favored adaptation in unusual environments and dissemination at long distances generating new biotypes.

Evaluation of biochemical and serological methods to identify and clustering yeast cells of oral Candida species by CHROMagar test, SDS-PAGE and ELISA.

The purpose of this work was to evaluate biochemical and serological methods to characterize and identify Candida species from the oral cavity. The strains used were five Candida species previously identified: C. albicans, C. guilliermondii, C. parapsilosis, C. krusei, C. tropicalis, and Kluyveromyces marxianus, as a negative control. The analyses were conducted through the SDS-PAGE associated with statistical analysis using software, chromogenic medium, and CHROMagar Candida (CA), as a differential medium for the isolation and presumptive identification of clinically important yeasts and an enzyme-linked immunoabsorbent assay (ELISA), using antisera produced against antigens from two C. albicans strains. This method enabled the screening of the three Candida species: C. albicans, C. tropicalis, and C. krusei, with 100% of specificity. The ELISA using purified immunoglobulin G showed a high level of cross-reaction against protein extracts of Candida species. The SDS-PAGE method allowed the clustering of species-specific isolates using the Simple Matching coefficient, S(SM) = 1.0. The protein profile analysis by SDS-PAGE increases what is known about the taxonomic relationships among oral yeasts. This methodology showed good reproducibility and allows collection of useful information for numerical analysis on information relevant to clinical application, and epidemiological and systematical studies.

Evaluation of biochemical and serological methods to identify and clustering yeast cells of oral Candida species by CHROMagar test, SDS-PAGE and ELISA

Este trabalho teve o propósito de avaliar métodos bioquímicos e sorológicos para serem aplicados na caracterização e identificação de linhagens do gênero Candida isoladas da cavidade bucal. As cepas empregadas representam cinco espécies de Candida previamente identificadas: C. albicans, C. guilliermondii, C. parapsilosis, C. krusei e C. tropicalis, utilizando como controle negativo Kluyveromyces marxianus. Foram empregadas as técnicas de gel de poliacrilamida (SDS-PAGE) associado à análise estatística em software, CHROMagar Candida (CA), meio cromogênico diferencial descrito para o isolamento e identificação presuntiva de leveduras de importância clínica e um ensaio de imunoabsorção ligado a enzima (ELISA), utilizando antissoro produzido contra extratos protéicos de uma linhagem-padrão de Candida e um isolado de cavidade oral de C. albicans. O método mostrou-se adequado para a identificação presuntiva de C. albicans, C. tropicalis e C. krusei, com 100% de sensibilidade e especificidade, com base na coloração e textura das colônias. O método de ELISA utilizando imunoglobulinas G purificadas apresentou alto teor de reação cruzada com as outras espécies de Candida estudadas. A análise do perfil protéico por SDS-PAGE permitiu agrupar os isolados da cavidade oral por intermédio do coeficiente "Simple Matching", SSM = 1,0. Os perfis protéicos analisados por SDS-PAGE ampliam os conhecimentos sobre as relaçäes taxonômicas de leveduras isoladas da cavidade oral. Esta metodologia demonstra boa reprodutibilidade e origina informaçäes úteis para aplicação clínica e estudos que envolvem a sistemática e a epidemiologia.

Psicoterapia psicodinâmica breve: estudo de acompanhamento / Brief psychodynamic psychotherapy: a follow-up study

Foram verificadas mudanças na adaptação e em variáveis psicodinâmicas ao final e no acompanhamento, após seis meses, de psicoterapias psicodinâmicas breves. N=33 (N = 23 mulheres, N = 10 homens, idades entre 21 e 60 anos, média 34,3). Apesar da maioria dos sujeitos apresentar adaptação não eficaz severa no início, as terapias causaram mudanças psicodinâmicas e adaptativas, em parte da amostra, que ainda estavam presentes seis meses após seu término. Mudanças nas estratégias devem ser buscadas, visando os mais comprometidos (AU)

Psicoterapia psicodinâmica breve: estudo de acompanhamento / Brief psychodynamic psychotherapy: a follow-up study

Verifica mudanças na adaptaçäo e em variáveis psicodinâmicas ao final e no acompanhamento, após 6 meses, de psicoterapias psicodinâmicas breves N=33 (N= 23 mulheres, N=10 homens, idades entre 21 e 60 anos, média 34,3). Apesar da maioria dos sujeitos apresentar adaptaçäo näo eficaz severa no início, as terapias causaram mudanças psicodinâmicas e adaptativas, em parte da amostra, que ainda estavam presentes seis meses após seu término. Mudanças nas estratégias devem ser buscadas, visando os mais comprometidos