Effect of Vitrification of Feline Ovarian Cortex on Follicular and Oocyte Quality and Competence

Cryopreservation of ovarian cortex has important implications in the preservation of fertility and biodiversity in animal species. Slow freezing of cat ovarian tissue resulted in the preservation of follicular morphology and in the follicular development after xenografting. Vitrification has been recently applied to ovarian tissues of different species, but no information is available on the effect of this method on feline ovarian cortex. Moreover, meiotic competence of fully grown oocytes isolated from cryopreserved tissue has not been reported. The aim of this study was to evaluate the effect of vitrification of feline ovarian cortex on follicular morphology and oocyte integrity, as well as meiotic competence. A total of 352 fragments (1.52 mm3) were obtained from ovarian cortical tissues: 176 were vitrified and 176 were used fresh as control. Histological evaluation of fresh and vitrified fragments showed intact follicles after cryopreservation procedures with no statistically significant destructive effect from primordial to antral follicles. After IVM, oocytes collected from vitrified ovarian fragment showed a higher proportion of gametes arrested at germinal vesicle (GV) stage compared to those isolated from fresh control tissue (33.8% vs 2.9%; p < 0.001). However, oocytes isolated from vitrified tissues were able to resume meiosis, albeit at lower rate than those collected from fresh tissues (39.8% vs 85.9%; p < 0.00001). Vitrification induced changes in the organization of cytoskeletal elements (actin microfilaments and microtubules) of oocytes, but significantly only for actin network (p < 0.001). Finally, chromatin configuration within the GV was not affected by the cryopreservation procedure. Our study demonstrated that vitrification preserves the integrity of ovarian follicles and that oocytes retrieved from cryopreserved tissue maintain the capability of resuming meiosis. To our knowledge, this has not previously been reported in the cat.

Hematological values and body, heart and liver weights of male and female broiler embryos of young and old breeder eggs

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Co-accumulation of microbial residues and particulate organic matter in the surface layer of a no-till Oxisol under different crops

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Terrain forms and spatial variability of soil properties in an area cultivatedwith citrus

A técnica de agricultura de precisão e a relação solo-paisagem permitem delimitar áreas para o manejo localizado, o que permite a aplicação localizada de insumos agrícolas e, consequentemente, pode contribuir para a preservação de recursos naturais. Portanto, o objetivo deste trabalho foi caracterizar a variabilidade espacial das propriedades químicas e do teor de argila, no contexto da relação solo-paisagem, em um Latossolo sob cultivo de citros. Amostras de solo foram coletadas na profundidade de 0,0-0,2 m, em uma área de 83,5 ha cultivada com citros, na forma de malha, com intervalos regulares de 50 m, com 129 pontos na forma de relevo côncava e 206 pontos na forma plana, totalizando 335 pontos. Os valores obtidos para as variáveis que expressam as propriedades químicas e para o teor de argila do solo foram submetidos à análise estatística descritiva e geoestatística com a modelagem de semivariogramas para a confecção de mapas de krigagem. Os valores de alcance e mapas de krigagem indicaram maiores variabilidades na forma de relevo côncava (segmento topo), quando comparada com a forma plana (segmentos meia encosta e encosta inferior). A identificação de diferentes formas de relevo mostrou-se eficiente no entendimento da variabilidade espacial das propriedades químicas e do teor de argila do solo sob cultivo de citros.

Modified 'Stent-Graft Sandwich' Technique for Treatment of Isolated Common Iliac Artery Aneurysm in Patient With Marfan Syndrome

Isolated iliac artery aneurysms are rare in the general population (0.03%) and represent 2% of all abdominal aneurysms, and the association with Marfan syndrome is even rarer. We report a Marfan syndrome case with an isolated common iliac artery aneurysm treated by using a modified 'stent-graft sandwich' technique, with preservation of the internal iliac artery perfusion. The modified 'stent-graft sandwich' technique involves building an appropriate proximal neck just in the common iliac artery for fittingly housing two new stent-grafts inside, both deployed simultaneously and each one going to both distal iliac arteries (internal and external).

Ultrasound guidance is not necessary during easy embryo transfers

Purpose: To determine whether the use of ultrasound (US) to guide embryo transfer (ET) in a population previously defined as likely to have easy transfer would change the implantation and pregnancy rates in an ICSI program.Methods: A total of 100 patients identified as likely to have easy transfer after mock transfer were divided into two groups: Group I, US-guided ET (N = 50) and Group II, ET without the aid of US (N = 50).Results: Implantation and pregnancy rates were similar (p = 0.51, p = 0.29) for Group I (19.6%,42%) and Group II (16.3%,30%), as also was the abortion rate (p = 0.55) (Group I: 1/21; Group II: 2/15).Conclusion: As long as previous mock transfers are routinely performed during a cycle preceding assisted reproduction and the clinician considers transfer to be easy, ultrasound does not benefit the process of embryo transfer.

Zona thinning with a noncontact diode laser in ICSI embryos from women of advanced age

Purpose: the objective of this study was to determine if the zona thinning (ZT) technique improved the rates of implantation and clinical pregnancy for patients aged, greater than or equal to38 years submitted to an ICSI program.Methods: A total of 100 patients submitted to ICSI and aged, greater than or equal to38 years were divided in a prospective and randomized manner into two groups: Group I - patients submitted to ZT (n = 50); a laser diode with 1.48 mum wavelength (Fertilaser) was used for the ZT procedure with 1-2 irradiations of 10 ms applied to four different positions on the zona pellucida (ZP) of each embryo to thin 60-90% of the ZP (each point with a 15-20 mum length of ZT). Group II - patients with no ZT (n = 50). In both groups, embryo transfer was performed on the second or third day.Results: the age of Group I patients (39.8 +/- 1.3) did not differ (p = 0.67) from that of Group II patients (40 +/- 1.9). The number of oocytes retrieved at metaphase II from Group I (6.4 +/- 4.2) and Group II (6.8 +/- 5) was similar (p = 0.94). Normal fertilization rates and cleavage rates were similar (p = 0.78 and p = 0.63, respectively) for Group I (71.5 +/- 22% and 96.7 +/- 11%) and Group II (73.5 +/- 19.7% and 96 +/- 11%, respectively). The number of embryos transferred was similar (p = 0.53) for the two groups (Group I = 3.1 +/- 1.3; Group II = 2.9 +/- 1.1). The thickness of the ZP of Group I embryos (16.9 +/- 2.4 mum) did not differ (p = 0.97) from that of Group II embryos (16.9 +/- 2.3 mum). The rates of embryo implantation and clinical pregnancy per embryo transfer were similar (p = 0.67, p = 0.61) for Group I (7 and 16%, respectively) and for Group II (8.2 and 22%, respectively).Conclusions: These results suggest that ZT in the population aged, 38 years may have no impact on ICSI success rates. However, this conclusion is limited to a situation in which length of the laser ZT was less than or equal to 20 mum and the laser was applied to four different positions.

Development of a real-time PCR method for rapid sexing of human preimplantation embryos

Genes on the X chromosome are known to be responsible for more than 200 hereditary diseases. After IVF, the simple selection of embryo sex before uterine transfer can prevent the occurrence of affected offspring among couples at risk for these genetic disorders. The aim of this investigation was to develop a rapid method of preimplantation genetic diagnosis (PGD) using real-time polymerase chain reaction (PCR) for the sexing of human embryos, and to compare it to the fluorescence in-situ hybridization technique, considered to be the gold standard. After biopsies were obtained from 40 surplus non-viable embryos for transfer, a total of 98 blastomeres were analysed. It was possible to analyse 24 embryos (60%) by both techniques, generating a total of 70 blastomeres (35 per technique), white 28 blastomeres from 16 embryos (40%) were analysed only by real-time PCR. A rapid and safe method was developed in the present study for the sexual diagnosis of a single human cell (blastomere and buccal cell) using the emerging technology of real-time PCR. (C) 2009, Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

Cold Renal Ischemia: Comparison of Efficacy Between Two Techniques of Cooling, in a Swine Model

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Factors potentially affecting fertility of lactating dairy cow recipients

Objectives of this study were to evaluate factors that could affect pregnancy rate after embryo transfer (ET) in lactating dairy cow recipients. The trial was conducted at a dairy farm located in Descalvado, SP, Brazil from October 2003 to September 2004. From 1037 cows with CL that were treated with an injection of PGF(2 alpha), 43.3% were detected in heat; 263 were previously assigned at day of PGF(2 alpha) injection for AI and 186 for ET. Ovulation rate was 85.7% (385/449). Pregnancy rate for cows with CL for AI and embryo transfer recipients were 36.5% (84/230) and 58.7% (91/155) at day 25 and 33.0% (76/230) and 45.8% (71/155) at day 46, respectively. Embryonic loss were 9.5% (8/84) for the AI group and 21.9% (20/91) for the ET group. Average milk production was 31.4 L/day/cow. Average daily milk production from 7 days before PGFZ injection to 7 days after ET tended (P < 0.10) to influence pregnancy rate on days 25 and 46. Average daily milk production from the day of embryo transfer to 7 days after influenced embryonic loss (P < 0.05). Cows with higher milk production had lower probability of pregnancy and higher probability of embryonic loss. Cows with higher days in milk had higher probability of pregnancy. Cows with higher rectal body temperature had lower probability of pregnancy and higher probability of embryonic loss. The influence of high milk yield and body temperature on fertility in lactating dairy cow recipients suggests that these effects can occur also after embryo reaches the blastocyst stage. (c) 2005 Elsevier B.V. All rights reserved.

Comparison of in vitro and in vivo fertilizing potential of bovine semen frozen in egg yolk or new lecithin based extenders

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Lipid content and apoptosis of in vitro-produced bovine embryos as determinants of susceptibility to vitrification

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Tooth replantation after use of euro-coffins solution or bovine milk as storage medium: a histomorphometric analysis in dogs

Purpose: Euro-Collins solution was developed for the preservation of organs for transplantation, whose characteristics have raised interest for its use as a storage medium for avulsed teeth before replantation. This study evaluated histologically and morphometrically the healing process of dog teeth replanted after storage in Euro-Collins solution or bovine milk.Materials and Methods: Eighty roots of 4 young adult mongrel clogs were randomly assigned to 4 groups (n = 20) and the root canals were instrumented and obturated with gutta-percha and a calcium hydroxide-based sealer. After 2 weeks, the teeth were extracted and subjected to the following protocols: GI (negative control), replantation immediately after extraction; GII (positive control), bench-drying for 2 hours before replantation; GIII and GIV, immersion in 10 mL of whole bovine milk and Euro-Collins solution at 4 C, respectively, for 8 hours before replantation. The animals were sacrificed 90 days postoperatively. The pieces containing the replanted teeth were subjected to routine processing for histologic and histometric analyses under light microscopy and polarized light microscopy.Results: Root resorption was observed in all groups. GII exhibited the greatest loss of dental structure (P < .01), and inflammatory resorption was predominant in this group. Storage in milk showed poorer results than immediate replantation and storage in Euro-Collins solution (P < .01). The teeth stored in Euro-Collins solution presented similar extension of root resorption and periodontal ligament reorganization to those of immediately replanted teeth.Conclusions: The findings of this study suggest that the Euro-Collins solution is an adequate storage medium for keeping avulsed teeth for up to 8 hours before replantation.

Treatment of gingival fibromatosis associated with Zimmermann-Laband syndrome

Background: Gingival fibromatosis is a rare condition characterized by a generalized enlargement of the buccal and lingual aspects of the attached and marginal gingiva.Methods: This case report describes the periodontal management of a 13-year-old female patient with gingival fibromatosis associated with Zimmermann-Laband syndrome. The patient presented with gingival enlargement involving the maxillary and the mandibular arches, anterior open bite, and non-erupted teeth. Periodontal treatment included gingivectomy in all four quadrants.Results: Histopathologic evaluation of the excised tissue supported the diagnosis of gingival flbromatosis. A significant improvement in esthetic appearance and eruption of the non-erupted teeth were obtained. The patient was referred for appropriate orthodontic treatment and has been closely followed for the earliest signs of recurrence of gingival enlargement.Conclusions: the successful therapy for gingival fibromatosis depends on correctly identifying the etiological factors and improving the impaired function and esthetic appearance through surgical intervention and adjunctive orthodontics. Maintaining treatment results depends on preservation of periodontal health.

Reproduction of Pirapitinga do Sul (Brycon opalinus Cuvier, 1819) in the Parque Estadual da Serra do Mar-Núcleo Santa Virgínia, São Paulo, Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)