The aim of this study was to carry out in vitro fertilization using spermatozoa selected with Androcoll-E™ and to evaluate the efficiency of the culture medium DMEM-F12 for in vitro embryo development in the llama. Twelve adult females from 18 superstimulated (67%) were used as oocyte donors. They were superstimulated with 1500 IU of eCG and after 5 days, received a single dose of buserelin. Twenty hours post-injection, follicular aspiration was conducted by flank laparotomy. Semen collections were performed under general anesthesia by electroejaculation of the male. The ejaculates were processed with a solution of collagenase (0.1%) and an Androcoll-E™ column was used to improve the sample. Sixty nine COCs were recovered from 79 aspirated follicles (87% recovery). Only expanded COCs were used (n = 67); they were randomly placed in groups of 1-5 in Fertil-TALP and the sperm suspension (20 × 10(6) live spermatozoa/ml) was added to each fertilization microdroplet. After 24 h, they were randomly placed in one of two culture media: SOF (n = 34) or DMEM-F12 (n = 33) and incubated for 6 days in humidified atmosphere of 5% CO(2) , 5% O(2) and 90% N(2) at 38°C. The blastocyst rate was 20% (7/34) in SOF medium (3 hatched, 2 expanded and 2 early blastocysts) and 15% (5/33) in DMEM medium (all expanded blastocysts). In conclusion, using Androcoll-E™ it is possible to select good quality spermatozoa from llama ejaculates for in vitro fertilization and to produce blastocysts in DMEM-F12 medium. This is also the first time that hatched llama blastocysts have been produced after culture in a defined medium such as SOFaa.
The aim of this study was to evaluate the developmental competence and pregnancy rate of llama hatched blastocysts produced in vitro using gametes from live animals and two different culture conditions. Fifteen adult females were superstimulated with 1500 IU of eCG, eleven (73%) responded to the treatment and were used as oocyte donors. Follicular aspiration was conducted by flank laparotomy. Semen collections were performed under general anesthesia by electroejaculation of the male. Sixty-six COCs were recovered from 77 aspirated follicles (86% recovery) and were randomly placed in Fertil-TALP microdroplets with the sperm suspension (20 × 10(6)live spermatozoa/ml). After 24 h, they were placed in SOFaa medium supplemented with FCS and randomly assigned to one of two culture conditions. Culture condition 1 (CC1) consisted of 6 days of culture (n=28) and culture condition 2 (CC2) consisted of renewing the culture medium every 48 h (n=35). In CC1, the blastocyst rate was 36% (10/28) and the hatched blastocyst rate was 28% (8/28) whereas in CC2, the blastocyst rate was 34% (12/35) and the hatched blastocyst rate was 20% (7/35) (p>0.05). No pregnancies were obtained after embryo transfer (ET) of CC1 blastocysts (0/8) while one pregnancy was obtained (1/7) after transferring a hatched blastocyst from CC2. Forty-two days after the ET, the pregnancy was lost. This study represents the first report of a pregnancy in the llama after intrauterine transfer of embryos produced by in vitro fertilization using gametes from live animals.
Seminal plasma is intimately connected to sperm physiology and particularly in South American Camelids, has demonstrated to be involved in multiple physiological reproductive events. Different percentages of seminal plasma (0%, 10% and 50%) were added to thawed llama semen samples with the objective of evaluating the interaction with cryopreserved sperm over time (0, 1.5 and 3 hr at 37°C). A total of 20 ejaculates from five adult llama males (n = 5; r = 4) were evaluated. A significant decrease in sperm motility, membrane function and live sperm was observed in all thawed samples (0%, 10% and 50%) at 0 hr when compared to raw semen. Neither morphology nor chromatin condensation was altered in all thawed samples (p > .05), but a significant increase in the percentage of spermatozoa with fragmented DNA was observed after thawing all samples versus raw semen. When evaluating thawed samples over time, a significant decrease of motility and membrane function was observed, while the percentages of total live sperm were preserved over the 3 hr of incubation in all final concentrations evaluated. To conclude, the addition of 10% or 50% of seminal plasma was incapable of preserving motility or membrane function of frozen‐thawed llama sperm during 3 hr of incubation.
Objective: To determine the initial management and in-hospital mortality of patients with acute coronary syndrome who attended referral hospitals in Paraguay. Method: Observational, multicenter study, in patients over 18 years with a confirmed diagnosis of acute coronary syndrome. Results: 780 patients were included from May 2015 to February 2016; the mean age was 64.1 ± 12.3 years, 64.1% male. The clinical presentation were acute coronary syndrome with ST elevation in 40.1% and without elevation in 59.9%. In patients with ST elevation there is a high percentage of late attendance, more than 12 h of evolution in 49.8%; those with less than 12 h of evolution underwent reperfusion in 52.2% of the cases, received fibrinolytics in 36.3% of the cases, and primary percutaneous coronary intervention 15.9%. In-hospital mortality for acute coronary syndrome was 10.3%, with ST-segment elevation was 12.8%, and without ST-segment elevation was 8.6%. Conclusions: The management of acute coronary syndrome in Paraguay needs a comprehensive approach, which promotes earlier care, and increases the implementation of reperfusion therapies in the health services network, in order to improve the therapeutic response rates and decrease hospital mortality.
Objective: Breastfeeding is associated with multiple benefits for preterm infants. Kangaroo care is a strategy geared towards making the aforementioned practice successful. This study examined the rate of exclusive breastfeeding (EBF) at the 30th day of corrected gestational age (CGA) appointment and the maternal and neonatal factors associated with this outcome in preterm infants assisted by kangaroo care. Methods: A cross-sectional study was conducted on very low birth weight (below 1500g) newborns and/or those with gestational age below 33 weeks, born from January 2019 to February 2020 and monitored in the outpatient follow-up. For EBF yes (G1) or no (G2), bivariate and multivariate analyses were performed. Results: 87 (100%) patients attended the 30th day of CGA appointment, 43 (49.4%) on EBF, 30 (34.5%) on breastfeeding and 14 (16.1%) on formula only. The median gestational age and birth weight were 31 weeks (29-32) and 1370g (1190-1670), respectively, and 51 (58.6%) patients were female. There was no difference in the bivariate analysis of maternal age, preeclampsia, route of delivery, antenatal corticosteroid use, birth weight, neonatal morbidities, length of hospitalization, chronological age, and anthropometry at the 30th day of CGA between the G1 and G2 groups. In multivariate analysis, gestational age above 30 weeks (RC3.11/p=0.034) and breastfeeding duration above 30 minutes (RC3.77/p=0.013) were determinants of EBF. Conclusion: Successful breastfeeding rates were observed in 2020 in this at-risk population, especially compared to previous years' results from this and other institutions. The newborn's longer breastfeeding time, one of the strategies of kangaroo care, was shown to be a protective factor for the maintenance of EBF in this outpatient follow-up, and special attention should be given to those born under 30 weeks, who showed a higher risk of weaning.
Objetivo: Determinar el tratamiento inicial y la mortalidad intrahospitalaria de pacientes con síndrome coronario agudo que acudieron a centros hospitalarios de referencia de Paraguay. Método: Estudio observacional y multicéntrico en pacientes mayores de 18 años con diagnóstico confirmado de síndrome coronario agudo. Resultados: Se incluyó a 780 pacientes desde mayo de 2015 hasta febrero de 2016; la edad media fue de 64.1 ± 12.3 años y el género masculino representó el 64.1%. La presentación clínica fue la de síndrome coronario agudo con elevación del ST en 40.1% y sin elevación del ST en 59.9%. En pacientes con elevación del ST se observó un alto porcentaje de consultas tardías, mayor de 12 h de evolución en 49.8%; en aquéllos con menos de 12 h de evolución se indicó la reperfusión en 52.2%, el 36.3% recibió fibrinolíticos y 15.9% intervención coronaria percutánea primaria. La mortalidad hospitalaria del síndrome coronario agudo fue de 10.3%, con elevación del segmento ST en 12.8% y sin elevación del segmento ST en 8.6%. Conclusiones: El tratamiento del síndrome coronario agudo en el Paraguay requiere un abordaje integral, que promueva consultas más tempranas y aumente la institución de tratamientos de reperfusión en la red de servicios de salud; el objetivo es mejorar los índices de respuesta terapéutica y disminuir la mortalidad hospitalaria.
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