RESUMO
In this study we examined quality, longevity and ability of epididymal sperm (EP) to bind to oviduct explants (OE) after cooling and cryopreservation. Ejaculated (EJ) and EP sperm from seven bulls were evaluated before, during and after cryopreservation for total (TM), progressive motility (PM), sperm morphology, plasma membrane integrity (PMI) and acrosome integrity (ACI). For longevity, cryopreserved EP, EJ and a third group of cells in which EP spermatozoa were incubated with seminal plasm (SP) for 10 min after thawing (EPP group), were compared, and the groups were analyzed at 0, 3, 6, and 24 h for all parameters. Sperm from each group were co-incubated with OE for 30 min, 6 h, and 24 h for binding evaluation. Data were analyzed by the generalized linear models SAS 9.1 (P < 0.05). After cooling, EP displayed higher TM, higher PMI, and higher ACI (P < 0.05) than EJ. No differences were noted in the percentage of spermatozoa with PMI and AI between EJ and EP for fresh, cooled or cryopreserved sperm. However, a reduction in motility occurred in the EJ sperm after cooling, while in EP group such reduction occurred only after cryopreservation. At 6 h of incubation EP and EPP had higher PMI and ACI than EJ (P < 0.05). The number of spermatozoa bound to OE was similar P > 0.05) for all groups either at 30 min or 24 h. We conclude that EP are more resistant to cooling than EJ, and can bind to OE similarly to EJ.
Assuntos
Criopreservação/métodos , Epididimo/citologia , Preservação do Sêmen/métodos , Espermatozoides/citologia , Animais , Bovinos , Ejaculação , Masculino , Motilidade dos Espermatozoides/efeitos dos fármacos , Espermatozoides/efeitos dos fármacosRESUMO
INTRODUCTION: Non-pharmacological intervention options, including individual reminiscence therapy (iRT), have been effective in improving cognitive functioning, mood, and quality of life (QoL) in persons with neurocognitive disorders (NCD). OBJECTIVES: A 13-week randomized trial intervention utilizing iRT was conducted on older adults with NCD. We explored predictors of participants with positive and non-positive intervention responses using responder analysis, an analytic strategy that focuses on contributors to intervention response. PATIENTS AND METHODS: Re-analysis of a published single-blind, multicentre, randomised controlled trial on 251 older adult residents with NCD from residential facilities across Portugal. Participants received 13 weeks of biweekly iRT (26 sessions) or treatment/programming as usual. Outcomes included global cognition (Minimental State Examination), memory (MAT), executive functioning (FAB), depressive symptoms (GDS-15), and QoL (QoL-AD). RESULTS: There were more responders in the intervention than the control group on all five criteria, with significant differences for cognition (p = 0.001; f = 0.202; NNT = 5) and memory (p = 0.004; f = 0.184; NNT = 6). At baseline, intervention responders vs non-responders had: higher QoL-AD scores (30.23 vs 25.57; p < 0.001; d = -0.774) for cognition; lower FAB scores (1.41 vs -2.12; p < 0.001; d = 0.928) for executive functioning; higher GDS-15 scores for the depressive symptoms (7.57 vs 4.91; p < 0.001; d = -0.845), and for QoL (6.81 vs 5.33; p = 0.013; d = -0.443). CONCLUSIONS: The iRT intervention showed high response rates for cognition and memory. Those with worse executive dysfunction, mood, and QoL, benefitted most from the intervention for those respective outcomes. Therefore, the presented iRT has beneficial effects for people with NCD, with mood and QoL as important influential factors.
TITLE: Ensayo aleatorio de terapia de reminiscencia individual para adultos mayores con deterioro cognitivo: un análisis de respuesta de tres meses.Introducción. La terapia de reminiscencia individual (iTR) ha demostrado mejorar la cognición, el estado de ánimo y la calidad de vida (CdV) de personas con trastornos neurocognitivos (TNC). Objetivo. Se exploraron los predictores de la respuesta positiva a la iTR utilizando el análisis de respuesta, una estrategia analítica de los factores que contribuyen a una respuesta a la intervención. Pacientes y métodos. Reanálisis de un ensayo controlado aleatorizado de 251 adultos mayores portugueses con TNC. Los participantes recibieron dos sesiones de iTR durante 13 semanas (26 sesiones) o el tratamiento habitual. Las variables analizadas fueron la cognición global (Minimental State Examination), la memoria (test de alteración de la memoria), el funcionamiento ejecutivo batería de evaluación frontal (FAB), los síntomas depresivos escala de depresión geriátrica-15 (GDS-15) y la CdV escala de calidad de vida en la enfermedad de Alzheimer (QOL-AD). Resultados. Hubo más respondedores en el grupo de intervención que en el de control en los cinco criterios, con diferencias significativas para cognición (p = 0,001; f = 0,202; número necesario para tratar = 5) y memoria (p = 0,004; f = 0,184; número necesario para tratar = 6). En la línea de base, los respondedores tenían: puntuaciones más altas de QOL-AD (30,23 frente a 25,57; p menor de 0,001; d = 0,774) para la cognición; puntuaciones FAB más bajas (1,41 frente a 2,12; p menor de 0,001; d = 0,928) para el funcionamiento ejecutivo; y mayores puntuaciones en la GDS-15 para los síntomas depresivos (7,57 frente a 4,91; p menor de 0,001; d = 0,845) y para la CdV (6,81 frente a 5,33; p = 0,013; d = 0,443). Conclusiones. La iTR mostró altas tasas de respuesta para la cognición y la memoria. Los que tienen peor función ejecutiva, estado de ánimo y CdV se beneficiaron más de la intervención para esas respectivas variables. La iTR tiene efectos beneficiosos en los TNC, con el estado de ánimo y la CdV como factores influyentes.
Assuntos
Disfunção Cognitiva/terapia , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Memória , Psicoterapia , Método Simples-Cego , Fatores de Tempo , Resultado do TratamentoRESUMO
During embryogenesis, one of the two X chromosomes is inactivated in embryos. The production of embryos in vitro may affect epigenetic mechanisms that could alter the expression of genes related to embryo development and X chromosome inactivation (XCI). The aim of this study was to understand XCI during in vitro, pre-implantation bovine embryo development by characterizing the allele-specific expression pattern of the X chromosome-linked gene, monoamine oxidase A (MAOA). Two pools of ten embryos, comprised of the 4-, 8- to 16-cell, morula, blastocyst, and expanded blastocyst stages, were collected. Total RNA from embryos was isolated, and the RT-PCR-RFLP technique was used to observe expression of the MAOA gene. The DNA amplicons were also sequenced using the dideoxy sequencing method. MAOA mRNA was detected, and allele-specific expression was identified in each pool of embryos. We showed the presence of both the maternal and paternal alleles in the 4-, 8- to 16-cell, blastocyst and expanded blastocyst embryos, but only the maternal allele was present in the morula stage. Therefore, we can affirm that the paternal X chromosome is totally inactivated at the morula stage and reactivated at the blastocyst stage. To our knowledge, this is the first report of allele-specific expression of an X-linked gene that is subject to XCI in in vitro bovine embryos from the 4-cell to expanded blastocyst stages. We have established a pattern of XCI in our in vitro embryo production system that can be useful as a marker to assist the development of new protocols for in vitro embryo production.
Assuntos
Blastocisto/fisiologia , Monoaminoxidase/genética , Inativação do Cromossomo X/genética , Alelos , Animais , Blastocisto/metabolismo , Bovinos , Células do Cúmulo , Metilação de DNA , Feminino , Perfilação da Expressão Gênica , Masculino , Monoaminoxidase/metabolismo , Fenótipo , Polimorfismo de Fragmento de Restrição , RNA Mensageiro/química , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNARESUMO
Male and female embryos are known to be different in developmental kinetics, metabolism, gene expression, and epigenetic patterns. Therefore, the objective of this study was to clarify whether the morphological criteria used to select embryos for cryopreservation lead to a deviation in the male:female ratio, and whether vitrification effects vary according to embryo sex. Initially, five sires were tested to evaluate the effect of the bull on embryo development, sex ratio, speed of development, and response to cryopreservation. Results showed that bulls affected (Pâ¯<â¯0.05) embryo production, response to cryopreservation, and sex ratio. Then, one bull was selected, and used to produce embryos in vitro to characterize the responses of male and female embryos to vitrification. Results suggested that male and female embryos have the same morphological responses to vitrification, as no differences (Pâ¯>â¯0.05) were observed between the two sexes in post-warming survival and re-expansion rates. However, their molecular responses as evaluated by gene expression (FOSL1, HSPB1, CASP3, CASP8, HSPA5, HSPA1A, G6PD, and PGK1) analysis indicated an effect of sex on vitrification; vitrified female embryos exhibited higher mRNA levels of HSPA1A, CASP3, and G6PD compared to their male counterparts. In conclusion, bulls affected embryo production, speed of development, sex ratio, and response to cryopreservation. Male and female embryos differed in their molecular responses to vitrification; and also, deviations in the male:female ratio when selecting embryos for cryopreservation were confirmed.
Assuntos
Blastocisto/fisiologia , Bovinos/embriologia , Criopreservação/veterinária , Técnicas de Cultura Embrionária/veterinária , Animais , Desenvolvimento Embrionário/fisiologia , Feminino , Masculino , Fatores Sexuais , VitrificaçãoRESUMO
The objective was to evaluate the effect of Percoll volume, and duration and force of centrifugation on sperm quality characteristics, embryo development, and sex ratio of in vitro-produced (IVP) bovine embryos. Frozen-thawed semen from four bulls were submitted to three Percoll procedures: T1-4 mL of Percoll, centrifuged for 20 min at 700 g; T2-800 microL of Percoll, centrifuged for 20 min at 700 g; and T3-800 microL of Percoll, centrifuged for 5 min at 5,000 g. Sperm total motility, morphology and integrity of the sperm acrosome, membrane and chromatin were determined before and after Percoll treatment, and semen was used for in vitro fertilization (IVF) of in vitro-matured oocytes. All Percoll methods increased the proportion of motile sperm (P<0.05). There were no significant effects of treatment for any sperm characteristic; however, for every end point, there were significant differences among bulls. Similarly, rates of cleavage and blastocyst formation were not affected by the Percoll procedure (P>0.05), but were affected by sire (P<0.05). Sex ratio was similar among treatments for Bulls 2 and 3, whereas semen from Bull 1 processed by T1 yielded a greater percentage of male embryos. However, when only treatments were considered, independent of bulls, the proportion of male:female embryos did not differ significantly from an expected 1:1 ratio. In conclusion, decreasing Percoll volume, reducing duration of centrifugation, and using a higher force of centrifugation did not significantly affect sperm quality, embryo development, or sex ratio of in vitro-produced bovine embryos.
Assuntos
Bovinos/embriologia , Fertilização in vitro , Hipergravidade , Povidona/farmacologia , Razão de Masculinidade , Dióxido de Silício/farmacologia , Animais , Centrifugação , Relação Dose-Resposta a Droga , Técnicas de Cultura Embrionária , Embrião de Mamíferos/citologia , Embrião de Mamíferos/efeitos dos fármacos , Embrião de Mamíferos/fisiologia , Feminino , Fertilização in vitro/métodos , Fertilização in vitro/veterinária , Hipergravidade/efeitos adversos , Masculino , Análise para Determinação do Sexo , Fatores de TempoRESUMO
Single crystal of monoglycine nitrate has been studied by Raman spectroscopy under high pressures up to 5.5 GPa. The results show changes in lattice modes in the pressure ranges of 1.1-1.6 GPa and 4.0-4.6 GPa. The first change occurs with appearance of bands related to the lattice modes as well as discontinuity in the slope of dΩ/dP of these modes. Moreover, bands associated with the skeleton of glycine suggest that the molecule undergoes conformational modifications. The appearance of a strong band at 55 cm(-1) point to a second phase transition associated with the lattice modes, while the internal modes remain unchanged. These anomalies are probably due to rearrangement of hydrogen bonds. Additionally, decompression to ambient pressure shows that the phase transitions are reversible. Finally, the results show that the nitrate anions play an important role on the stability of the monoglycine nitrate crystal.
RESUMO
The aim of this study was to evaluate the superovulatory (SOV) response of Sindhi (Bos indicus) donors submitted to an ovarian follicular superstimulatory protocol replacing the last two doses of pFSH by eCG. Forty-eight SOV treatments were performed in a crossover design in 19 nulliparous and primiparous females that were randomly divided into two groups: FSH (n=24), which consisted of eight pFSH injections, or FSH/eCG (n=24), which consisted of six pFSH injections followed by two eCG injections. Each female underwent two or three SOV treatments that consisted of an i.m. injection of 2mg estradiol benzoate and the insertion of an intravaginal progesterone-releasing device on Day 0. On Day 4, superstimulatory treatments were initiated and 100mg pFSH was divided into twice daily decreasing doses over a 4-day period. In the FSH/eCG group, the last two doses of pFSH were replaced by two doses of eCG (150 IU eCG each). At the time of the fifth and sixth injections of FSH, 0.150 mg PGF(2α) was injected i.m. The intravaginal progesterone-releasing device was removed at the time of the last FSH or eCG injection and ovulation was induced with 0.2 mg GnRH 18 h later. All females were artificially inseminated with frozen-thawed semen from the same bull 6 and 18 h after GnRH treatment. Seven days after GnRH treatment, embryos/ova were recovered and classified. Follicular superstimulatory (number of follicles ≥6mm at the time of the last FSH or eCG injection) and SOV (CL number) responses were determined by transrectal ultrasonography. Data were analyzed using generalized linear models and results were presented as least squares means±standard error. The FSH/eCG group had higher superstimulatory (33.8±3.9 compared to 23.8±2.6 follicles; P=0.03) and SOV (16.8±2.9 compared to 10.8±2.1 CL; P=0.10) responses. Although the number of total ova/embryos was not different between groups (8.2±1.8 compared to 5.9±1.4 for FSH/eCG and FSH groups, respectively; P=0.25), the number (5.8±1.3 compared to 2.6±0.7; P=0.02) and percentage (75.6±5.7 compared to 53.2±9.7%; P=0.05) of transferable embryos was greater for the FSH/eCG females. Therefore, there was improvement in follicular superstimulatory and SOV responses and embryo quality in FSH/eCG-treated females.
Assuntos
Bovinos/fisiologia , Hormônio Foliculoestimulante/administração & dosagem , Gonadotropinas Equinas/administração & dosagem , Folículo Ovariano/efeitos dos fármacos , Indução da Ovulação/veterinária , Superovulação/efeitos dos fármacos , Animais , Brasil , Estudos Cross-Over , Feminino , Inseminação Artificial/veterinária , Análise dos Mínimos Quadrados , Masculino , Folículo Ovariano/diagnóstico por imagem , Folículo Ovariano/fisiologia , Indução da Ovulação/métodos , Gravidez , Distribuição Aleatória , Superovulação/fisiologia , UltrassonografiaRESUMO
The objective was to evaluate the structural and functional quality of bull sperm after sexing by flow cytometry. Frozen non-sexed (NS), sexed for X (SX) and sexed for Y (SY) sperm from four bulls was used. Frozen-thawed sperm was analyzed for motility, sperm head agglutination, morphology, capacitation, and integrity of the plasma membrane, acrosome, and chromatin. After Percoll centrifugation (45:60% gradients), the pellet was used for sperm analysis or IVF. Data were analyzed using generalized linear models (P < 0.05) and were reported as least squares means ± standard error (SEM). Based on sperm evaluations, NS sperm had better (P < 0.05) quality than sexed sperm, including higher motility and greater percentages of cells with an intact membrane and acrosome (58.0 ± 3.0, 58.2 ± 3.0, and 60.9 ± 3.3) than SX (29.6 ± 1.3, 36.0 ± 2.9, and 37.1 ± 3.3), and SY (26.2 ± 2.1, 36.4 ± 2.9, and 37.5 ± 3.3). There were no differences (P > 0.05) among groups for fertilization and cleavage rates. Similarly, blastocyst rate on Day 8 (Day 0 = day of insemination) did not differ among groups (22.2 ± 3.2, 18.1 ± 3.3, and 14.8 ± 2.9 for NS, SX, and SY, respectively). Regarding embryo development kinetics, all groups had similar developmental stages from Days 6 to 9. Although the sex-sorting procedure affected sperm characteristics, it did not significantly affect fertilization or embryo development.