Biomedical subjects

Salvador Ruiz

Publications and source records attributed to Salvador Ruiz.

6 recordsLinked to original sources

PubMed2006

Analysis of different factors influencing the intracytoplasmic sperm injection (ICSI) yield in pigs.

Intracytoplasmic sperm injection (ICSI) in pigs is a technique with potential application in diverse fields of animal production and biomedicine. Even though there are some cases of live offspring resulting from this technique, its yield is still quite low compared to other species. The aim of this study was to evaluate different factors affecting the ICSI performance. This was done by studying (1) the sequence of culture media for the oocytes after injection; (2) modifications in the in vitro maturation system (IVM) through meiotic inhibitors such as roscovitine, and changes in the IVM time; (3) oocyte activation through injection of inositol triphosphate (InsP(3)) together with the sperm. In vitro matured oocytes were employed. All the ICSI experiments were performed with fresh ejaculated semen. Results showed that porcine ICSI zygotes give an improved proportion of two-cell embryos using the sequence IVF medium-embryo culture medium (NCSU-23) rather than transferring directly to NCSU-23. Pronuclear formation ability was not affected by prematuration, but a faster embryo development was observed in roscovitine treated oocytes. In relation to IVM times, oocytes matured for 36 h can achieve better fertilization percentages than oocytes matured for 44 h. These results were independent of the roscovitine treatment. Finally, no influence on embryo development was observed until the blastocyst stage with the use of the InsP(3) as an exogenous activating factor.

Animals↗

PubMed2005

Birth of piglets after transferring of in vitro-produced embryos pre-matured with R-roscovitine.

The objectives of this study were to evaluate: (1) the nuclear maturation, (2) the intracellular glutathione (GSH) content, (3) the normality of fertilization and (4) full development after transplantation of embryos derived from porcine oocytes pre-cultured with 50 micromol/l roscovitine (an inhibitor of p34cdc2/cyclin B kinase) for 22 h. After treatment with roscovitine, the nuclear configuration of oocytes (Hoechst staining) was comparable with those examined just after collection: the majority of oocytes were arrested at the germinal vesicle (GV) 1 stage (63.2%). Roscovitine-treated oocytes progressed through meiosis to the metaphase II stage in a conventional step-wise in vitro maturation (IVM) program for 44 h in a proportion similar to control ones (>85.0%). When roscovitine-treated oocytes and non-treated oocytes were matured for 44 h and then co-cultured with fresh spermatozoa for 18 h, no differences were observed in oocyte penetrability, proportion of monospermic penetration and male pronuclear formation (>87%). Roscovitine increased the GSH synthesis in oocytes at 22 h, whereas, after 44 h, roscovitine-treated oocytes had similar amounts of GSH to non-treated oocytes. Finally, surgical transfer of zygotes at 22-24 h post-insemination, derived from roscovitine-treated oocytes, resulted in one pregnancy with 12 piglets born; control non-treated zygotes resulted in one pregnancy and 10 piglets born. The full-term developmental ability of mammalian oocytes pre-cultured with roscovitine prior to IVM is thereby demonstrated. This validation is important before the introduction of roscovitine into routine procedures.

Animals↗

PubMed2005

Stereotactic noninvasive volume measurement compared with geometric measurement for indications and evaluation of gamma knife treatment.

OBJECT: Volume estimation is one of the most important criteria in the evaluation and follow up of radiosurgical treatments and outcomes; however, several limitations are involved in the calculation estimation of target volumes. METHODS: Retrospective and prospective studies were conducted to evaluate the efficacy of a new noninvasive stereotactic method when it is compared with geometric volume calculation of intracranial tumors for planning stereotactic radiosurgery treatment as well as for follow up and outcome evaluation. Two equations were created that permit comparison of the calculated and measured volumes. These equations took linear and quadratic forms, respectively. Volume estimation using the stereotactic approach compared with traditional volume calculation gave more accurate results regardless of the shape and size of the lesion. CONCLUSIONS: The use of stereotactic volume calculation is highly recommended in planning, follow up, and determination of the outcome in patients participating in radiosurgical treatment and should lead to more uniform reports of the response to treatment.

Brain Neoplasms↗

PubMed2004

Decrease in glutathione content in boar sperm after cryopreservation. Effect of the addition of reduced glutathione to the freezing and thawing extenders.

Although glutathione content in boar spermatozoa has been previously reported, the effect of reduced glutathione (GSH) on semen parameters and the fertilizing ability of boar spermatozoa after cryopreservation has never been evaluated. In this study, GSH content was determined in ejaculated boar spermatozoa before and after cryopreservation. Semen samples were centrifuged and GSH content in the resulting pellet monitored spectrophotometrically. The fertilizing ability of frozen-thawed boar sperm was also tested in vitro by incubating sperm with in vitro matured oocytes obtained from gilts. GSH content in fresh semen was 3.84 +/- 0.21 nM GSH/10(8) sperm. Following semen cryopreservation, there was a 32% decrease in GSH content (P < 0.0001). There were significant differences in sperm GSH content between different boars and after various preservation protocols (P = 0.0102 ). The effect of addition of GSH to the freezing and thawing extenders was also evaluated. Addition of 5 mM GSH to the freezing extender did not have a significant effect on standard semen parameters or sperm fertilizing ability after thawing. In contrast, when GSH was added to the thawing extender, a dose-dependent tendency to increase in sperm fertilizing ability was observed, although no differences were observed in standard semen parameters. In summary, (i) there was a loss in GSH content after cryopreservation of boar semen; (ii) addition of GSH to the freezing extender did not result in any improvement in either standard semen parameters or sperm fertilizing ability; and (iii) addition of GSH to the thawing extender resulted in a significant increase in sperm fertilizing ability. Nevertheless, future studies must conclude if this is the case for all boars. Furthermore, since addition of GSH to the thawing extender did not result in an improvement in standard semen parameters, this suggests that during the thawing process, GSH prevents damage of a sperm property that is critical in the fertilization process but that is not measured in the routine semen analysis.

Animals↗

PubMed2004

Use of stereotactic PET images in dosimetry planning of radiosurgery for brain tumors: clinical experience and proposed classification.

UNLABELLED: We developed a technique that allows the routine integration of PET in stereotactic neurosurgery, including radiosurgery. We report our clinical experience with the combined use of metabolic (i.e., PET) and anatomic (i.e., MRI and CT) images for the radiosurgical treatment of brain tumors. We propose a classification describing the relative role of the information provided by PET in this multimodality image-guided approach. METHODS: Between December 1999 and March 2003, 57 patients had stereotactic PET as part of their image acquisition for the planning of gamma knife radiosurgery. Together with stereotactic MRI and CT, stereotactic PET images were acquired on the same day using either (18)F-FDG or (11)C-methionine. PET images were imported in the planning software for the radiosurgery dosimetry, and the target volume was defined using the combined information of PET and MRI or CT. To analyze the specific contribution of the PET findings, we propose a classification that reflects the strategy used to define the target volume. RESULTS: The patients were offered radiosurgery with PET guidance when their tumor was ill-defined and we anticipated some limitation of target definition on MRI alone. This represents 10% of the radiosurgery procedures performed in our center during the same period of time. There were 40 primary brain lesions, 7 metastases, and 10 pituitary adenomas. Abnormal PET uptake was found in 62 of 72 targets (86%), and this information altered significantly the MRI-defined tumor in 43 targets (69%). CONCLUSION: The integration of PET in radiosurgery provides additional information that opens new perspectives for the optimization of the treatment of brain tumors.

Adolescent↗

PubMed2003

Use of the Leksell gamma knife C with automatic positioning system for the treatment of meningioma and vestibular schwannoma.

OBJECT: The authors report their experience using the Leksell gamma knife C (GK-C) for the treatment of meningioma and vestibular schwannoma (VS). METHODS: In December 1999, the first commercially available clinical GK-C was installed at the Université Libre de Bruxelles (Erasme Hospital, Brussels, Belgium). In January 2000, the system was upgraded and equipped with the automatic positioning system (APS). Between February 2000 and February 2003, the APS-equipped GK-C was used to perform 532 radiosurgical treatments, including those in 97 meningiomas and 101 VSs. Meningioma and VS represent 18 and 19%, respectively, of lesions in patients treated with GK-C at the authors' center. The mean number of isocenters per lesion was 9.5 (range 1-36): 18.1 (range 1-36) for meningioma and 12.8 (range 1-27) for VS. In 77.6% of the cases, the authors used a single helmet of collimators (55.5% in meningioma and 74.3% in VS). The most frequently used collimator size was 4 mm (46.7%). Whereas it was 4 mm in cases of VS (64.3%), it was 8 mm in cases of meningioma (41.6%). The APS could be used in 86% of the cases, either alone (79%) or in combination with trunnions (7%). There was a difference in the APS-based treatment success rate in meningiomas (85%) and VSs (94%). A significant difference was also noted in the conformity of the radiosurgical treatments between the two lesions. CONCLUSIONS: The APS-equipped GK-C represents an evolutionary step in radiosurgery. It requires adjustments by the treating team for its specific limitations, which vary among indications, as exemplified by the differences inherent between meningioma and VS in this series.

Humans↗