RESUMO
This work was designed to determine temperature conditions within the reproductive tract of the female pig and study their impact on ARTs. Temperatures were recorded using a laparo-endoscopic single-site surgery assisted approach and a miniaturized probe. Sows and gilts were used to address natural cycle and ovarian stimulation treatments, respectively. According to in vivo values, IVF was performed at three temperature conditions (37.0°C, 38.5°C and 39.5°C) and presumptive zygotes were cultured in these conditions for 20 h, while further embryo culture (EC) (21-168 h post-insemination) was maintained at 38.5°C. After 20 h, different fertility parameters were assessed. During EC, cleavage and blastocyst stages were evaluated. Sperm membrane fluidity at the experimental temperatures was studied by using differential scanning calorimetry and fluorescence recovery after photobleaching techniques. An increasing temperature gradient of 1.5°C was found between the oviduct and uterus of sows (P < 0.05) and when this gradient was transferred to pig in vitro culture, the number of poly-nuclear zygotes after IVF was reduced and the percentage of blastocysts was increased. Moreover, the temperature transition phase for the boar sperm membrane (37.0°C) coincided with the temperature registered in the sow oviduct, and sperm membranes were more fluid at 37.0°C compared with those of sperm incubated at higher temperatures (38.5°C and 39.5°C). These data suggest that there may be an impact of physiological temperature gradients on human embryo development.
Assuntos
Técnicas de Cultura Embrionária/métodos , Oviductos/fisiologia , Temperatura , Útero/fisiologia , Animais , Biomimética , Temperatura Corporal/fisiologia , Células Cultivadas , Técnicas de Cultura Embrionária/veterinária , Embrião de Mamíferos , Desenvolvimento Embrionário/fisiologia , Feminino , Fertilização in vitro/métodos , Fertilização in vitro/veterinária , SuínosRESUMO
STUDY QUESTION: Is O2 tension in the pig oviduct and uterus affected by the estrous cycle stage and the animal's age, and can the outcome of in vitro embryo development be improved by mimicking these physiological values? SUMMARY ANSWER: O2 tension within the pig reproductive organs is affected by the animal's age, and values close to those measured in vivo have a positive impact on embryo development and quality when used during IVF and embryo culture (EC). WHAT IS KNOWN ALREADY: To obtain a healthy embryo in vitro, it is necessary to adopt a culture microenvironment that approximates physiological conditions. Despite advances in surgical procedures and sensitive probes that allow accurate assessment of in vivo O2 tension, few such studies have been conducted recently in mammals. In addition, no reference values of physiological O2 tension in the reproductive tract exist for large animal models such as pig, and the effect of O2 tension on ART outcomes is unknown. STUDY DESIGN, SIZE, DURATION: This study was conducted in pigs. We measured oviductal and uterine O2 tension (n = 29 and 13, respectively) and then examined how the use of the physiological values in pig IVF and EC affected pig ART output (n = 1447 oocytes). PARTICIPANTS/MATERIALS, SETTING, METHODS: The oviductal and uterine O2 tension at the different stages of the estrous cycle was monitored using a laparo-endoscopic single-site surgery (LESS) assisted approach along with a flexible and thin miniaturized luminescent probe. Two groups of pigs, Large-white × Landrace breed, were used: for the first group, 16 pre-pubertal gilts (5 months old and 95 kg) were induced to ovulate with equine chorionic gonadotropin (eCG) and human chorionic gonadotropin (hCG); in the second group 13 mature sows (24-48 months and 185 kg) were used. IVF and EC were performed at two different O2 tensions: Atmospheric O2 (20%) and the mean in vivo value measured (7%). At 18-20 h post-insemination (hpi), a small sample of presumptive zygotes were fixed, stained and examined under epifluorescence microscopy to assess the fertilization rates. At 48 hpi, cleavage was evaluated under the stereomicroscope. Finally, at 180 hpi, development to the blastocyst stage was quantified, blastocyst morphology was assessed, and embryos were fixed and stained to count the mean cell number per blastocyst. MAIN RESULTS AND THE ROLE OF CHANCE: The mean O2 content within the pig oviduct and uterus was always lower than in ambient air. The average O2 percentage was higher in gilts (10.0%) than in sows (7.6%) (P < 0.0001). The cleavage rate of porcine in vitro fertilized embryos maintained under 7% O2 during IVF and EC was significantly higher (60.0 ± 2.3) compared with those cultured under 20% O2 (32.0 ± 2.2) (P < 0.05). An increase in the number of cells in embryos cultured under the low O2 concentration (88.9 ± 5.9) was observed compared to those cultured under 20% O2 (59.0 ± 5.0) (P < 0.05). LARGE SCALE DATA: None. LIMITATIONS, REASONS FOR CAUTION: Although minimally invasive surgery was used the effect of anesthesia and manipulations on O2 tension within the organs are unknown. WIDER IMPLICATIONS OF THE FINDINGS: Using physiological oxygen concentrations in IVF/EC could improve ART outcomes. STUDY FUNDING AND COMPETING INTEREST(S): This study was funded by Spanish Ministry of Economy and Competitiveness (MINECO) and European Regional Development Fund (FEDER). Grants AGL2012-40180-C03-01 and AGL2015-66341-R. The authors declare no conflict of interest.
Assuntos
Blastocisto/fisiologia , Desenvolvimento Embrionário/fisiologia , Oviductos/fisiologia , Oxigênio/fisiologia , Técnicas de Reprodução Assistida/veterinária , Útero/fisiologia , Animais , Feminino , Gravidez , SuínosRESUMO
We present our experience in the design and development of a training program in paediatric and neonatal laparoscopic surgery, and the determination of face validity by the attendants. Data included in the present study was obtained from five consecutive editions of our Neonatal and Paediatric Laparoscopic Surgery Course. Our training model, with a total duration of 21 hours, begins with acquisition of knowledge in ergonomics and instrument concepts, after which the attendants develop basic laparoscopic dexterity through the performance of hands-on physical simulator tasks. During the second and third days of the course, surgeons undertook various surgical techniques hands-on animal model. At the end of the training program, a subjective evaluation questionnaire was handed out to the attendants, in which different didactic and organizational aspects were considered. We obtained a highly positive score on all questions concerning the different topics and techniques included in the training program (> or = 9 points over 10). 78,5% of the 54 attendants was in accordance with the course total duration, whilst 21,5% considered that it should be of longer duration. Regarding abilities' self assessment, 79,1% considered themselves capacitated to perform trained procedures on live patients. The presented training model has obtained a very positive valuation score, leading to an increase in the attendants' self confidence in the application of learned techniques to their clinical practice.
Assuntos
Laparoscopia/educação , Modelos Educacionais , Pediatria/educação , Especialidades Cirúrgicas/educação , Currículo , Neonatologia/educaçãoRESUMO
INTRODUCTION/BACKGROUND: Reconstructive and ablative urologic techniques require special technical mastery, especially the intracorporeal suturing. OBJECTIVE: To report the subjective evaluation of a versatile ex-vivo model aimed to practice laparoscopic ureteric reconstructive techniques (LURT) on box-trainer. STUDY DESIGN: The model is a continuous portion of porcine urinary bladder ("dilated pelvis"), the vesico-ureteral joint ("stenosis") and healthy ureter. All 127 participants (n = 119 urologists and n = 8 paediatric surgeons) performed on the model laparoscopic Anderson-Hynes dismembered pyeloplasty, and then, in the animal model, different LURT procedures (ureteroneocystostomy, ureteric reimplantation and/or dismembered pyeloplasty). The model was subjectively evaluated (face and content validity), through a 12 items questionnaire, based on a Likert scale (1-5 points) and a global question (1-10 points). RESULTS: The total mean rating for 11/12 items was very high (>4points). Only one was rated under 3 points. The overall total mean rating from 1 to 10 points was very high (9.19 ± 0.82 points). In 10/12 items, expert's feedback (content validity) prevailed over non-experts (face validity). DISCUSSION/CONCLUSION: The model was highly accepted for the practice of LURT techniques. Additionally, it is cost-effective, easy to assemble, ethically considerate, and realistic.
Assuntos
Laparoscopia , Ureter , Obstrução Ureteral , Animais , Humanos , Pelve Renal , Suínos , Ureter/cirurgia , Obstrução Ureteral/cirurgia , Procedimentos Cirúrgicos UrológicosRESUMO
OBJECTIVE: The aim of this study is to prove the effectiveness of a low cost, artificial model for training of a laparoscopic urethrovesical anastomosis. MATERIALS AND METHODS: This study included urologists who attended specialised courses on laparoscopic radical prostatectomy (LRP) held during the period 2015 to 2017. They were divided into 2 groups according to their previous experience in laparoscopic surgery. The tasks performed on the artificial simulator were prostate resection, "task 1", and urethrovesical anastomosis, "task 2". Once these exercises were completed, the study participants filled in an anonymous questionnaire regarding their demographic data and experience level in laparoscopic surgery (LS). In addition, they gave their opinions about the didactic capacity of the artificial organ and evaluated its usefulness as a tool for LRP training. To demonstrate face and content validity, the participants judged the texture, consistency, morphology and evaluated its similarity to the real organ. The assessment was made with a five-point Likert scale. RESULTS: The students were divided into 2groups: 10 experts (Group E) and 12 novices (Group N). The only significant difference between the scores of novices and experts was regarding the inclusion of this tool in the training programs (Group E=5 points versus group N=4.4±0.59, P=.024). The experts' group rated all the items with higher scores than the novices' one. Regarding the general assessment of the simulation model, the novice participants gave an average score of 8.00±0.91 points out of 10, while the experts' group granted higher scores of 9.4±0,51. CONCLUSION: This artificial model has shown to have an elevated face, content and construct validity, as well being an optimal didactic tool for training in the techniques of prostate resection and laparoscopic urethrovesical anastomosis.
Assuntos
Laparoscopia/educação , Modelos Anatômicos , Prostatectomia/educação , Prostatectomia/métodos , Treinamento por Simulação , Uretra/cirurgia , Bexiga Urinária/cirurgia , Anastomose Cirúrgica , Humanos , MasculinoRESUMO
OBJECTIVE: To investigate the feasibility and safety of laparoscopy-assisted transvaginal resection of sigmoid cancer. METHODS: From 2009 to 2011 we performed 21 laparoscopy-assisted transvaginal resections of sigmoid cancers. RESULTS: The resected tumours were T1(6), T2(7) and T3(8). The median number of resected lymph nodes was 15. Median operative time was 150 min and no conversion to open technique was necessary. A total of eighteen patients tolerated a liquid diet 24 h after surgery. The average VAS pain score (0 = no pain, 10 = unbearable pain) was 1.8 after 24 h and 1.1 after 48 h. One patient experienced a postoperative ileus, two patients had vaginal spotting with serous flow and three patients suffered urinary tract infections. One patient developed a hernia at the umbilical trocar site that was operated on. None of the sexually active patients reported any differences in sexual activity pre-op and postop on a postoperative questionnaire. After a median of 25 (10-41) months of follow-up, all patients are alive and well, with no evidence of recurrent disease. CONCLUSION: Laparoscopy-assisted transvaginal resection of sigmoid cancer is feasible and safe in a selected group of patients.