44 results on '"Grillo JM"'
Search Results
2. Treatment of idiopathic light chain deposition disease: complete remission with bortezomib and dexamethasone.
- Author
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Souto Filho JT, Monteiro JM, and Andrade IB
- Subjects
- Humans, Male, Middle Aged, Remission Induction, Antineoplastic Agents therapeutic use, Bortezomib therapeutic use, Dexamethasone therapeutic use, Glucocorticoids therapeutic use, Immunoglobulin Light Chains, Paraproteinemias drug therapy
- Abstract
Light chain deposition disease (LCDD) is a rare clinical entity characterized by the deposition of light chain immunoglobulins in different tissues and primarily affects the kidneys, followed by the liver and heart. This disease often manifests as nephrotic syndrome with marked proteinuria and rapid deterioration of renal function. More than 50% of cases are secondary to multiple myeloma or other lymphoproliferative diseases, with a well-established treatment aimed at controlling the underlying disease. In rare cases, there is no detection of an associated hematological disease, referred to as idiopathic LCDD. In these cases, there is no evidence-based consensus on the therapeutic approach, and management is based on the clinical experience of reported cases. Here we report a case of idiopathic LCDD treated with bortezomib and dexamethasone with complete hematologic responses, significant reduction of proteinuria, and improved renal function.
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- 2016
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3. Laparoscopic ovariectomy by single-port access for ovarian cryopreservation.
- Author
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Courbiere B, Crochet P, Marcelli M, Saias-Magnan J, Grillo JM, and Agostini A
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- Adolescent, Adult, Feasibility Studies, Female, Humans, Postoperative Complications, Prospective Studies, Time Factors, Treatment Outcome, Cryopreservation, Laparoscopy methods, Ovariectomy methods
- Abstract
Purpose: The aim of this study is to assess the feasibility of ovariectomy by single-port access laparoscopy for cryopreservation., Methods: Observational prospective monocentric study including patients referred for an ovariectomy for ovarian tissue cryopreservation underwent ovariectomy by single-port access laparoscopy. Feasibility, intra- and post-operative complications, and quality of the ovarian tissue collected were reported., Results: Height patients were included. No conversion to standard laparoscopy or laparotomy was performed and no intra- or post-operative complications were reported. Median duration of surgery was 35 min (30-60). The quality of all the ovarian tissue collected was correct, and cryopreservation was possible for all patients., Conclusions: Ovariectomy for cryopreservation by laparoscopy with SPA seems feasible. The advantages of this technique are particularly interesting in these patients who require the least aggressive surgical technique possible and a rapid convalescence.
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- 2016
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4. [Association between sperm abnormalities and occupational environment among male consulting for couple infertility].
- Author
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Ould Hamouda S, Perrin J, Achard V, Courbière B, Grillo JM, and Sari-Minodier I
- Subjects
- Adult, Humans, Infertility, Male chemically induced, Male, Infertility, Male etiology, Occupational Exposure adverse effects, Spermatozoa pathology
- Abstract
Alteration of sperm parameters related to occupational exposures is the subject of several studies, often on a case-control approach. The study populations usually comprise men consulting in infertility clinics for couple infertility. The objective of this review is to identify, from these case-control studies, the main occupational factors that may be associated with altered sperm parameters. We selected 13 articles in the PubMed database. Participation in these studies varied from 61 to 2619 subjects, with great methodological heterogeneity, particularly in the characterization of exposure. The main occupations that appear significantly associated with a risk of altered sperm parameters are workmen, painters, farmers, welders, plumbers and technicians. When analysis focuses on occupational exposures, a significant result is reported for solvents, heavy metals, heat, vibrations and non-ionizing radiation. None of the selected studies has found a link with exposure to pesticides., (Copyright © 2015 Elsevier Masson SAS. All rights reserved.)
- Published
- 2016
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5. [Elective single embryo transfer policy at 48/72 h: which results after fresh transfers and frozen/thawed transfers?].
- Author
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Bottin P, Achard V, Courbiere B, Paci M, Gnisci A, Jouve E, Grillo JM, and Paulmyer-Lacroix O
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- Cryopreservation, Elective Surgical Procedures, Embryo Transfer methods, Female, France, Hot Temperature, Humans, Policy, Pregnancy, Pregnancy Rate, Pregnancy, Multiple, Time Factors, Treatment Outcome, Single Embryo Transfer methods
- Abstract
Objectives: To evaluate our elective single embryo transfer policy performed at 48/72h and define predictive factors of pregnancy after frozen/thawed embryo transfer., Methods: Analysis of 289 elective single embryo transfers (eSET) performed in a selected population in the ART center of Marseille University Hospital from January 2005 to December 2011, as well as the 325 following frozen/thawed embryo transfers performed in this population between May 2005 and December 2012., Results: Cumulative pregnancy rate/oocyte retrieval was of 62.6%; 45% of the couples obtained the birth of at least one child. During this studied period, cumulative pregnancy and delivery rates in the whole population remained stable while multiple delivery rate/delivery clearly decreased. Elective single embryo after frozen/thawed transfer gave satisfying results (24.6% pregnancy/transfer) only in the lack of lysis or in case of mild lysis (1-25%) of the transferred embryo., Conclusion: The implementation of an eSET policy gives satisfying results, depending largely on embryo quality. By proposing eSET to a well-targeted population, chosen both on clinical and biological criteria, a clear reduction of cumulative multiple delivery rate/delivery was obtained in our center over this period, without any global decrease of cumulative pregnancy rate/attempt. Embryo quality is a major factor of success, especially in frozen/thawed cycles. The elective single embryo frozen/thawed transfer should be carried out only if embryo lysis after thawing does not exceed 25%., (Copyright © 2015 Elsevier Masson SAS. All rights reserved.)
- Published
- 2015
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6. Nuclear envelope remodelling during human spermiogenesis involves somatic B-type lamins and a spermatid-specific B3 lamin isoform.
- Author
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Elkhatib R, Longepied G, Paci M, Achard V, Grillo JM, Levy N, Mitchell MJ, and Metzler-Guillemain C
- Subjects
- Acrosome metabolism, Acrosome ultrastructure, Adult, Animals, Cell Differentiation, Gene Expression Regulation, HeLa Cells, Humans, Lamin Type A genetics, Lamin Type A metabolism, Lamin Type B genetics, Male, Mice, Nuclear Lamina ultrastructure, Protein Isoforms genetics, Protein Isoforms metabolism, RNA, Messenger genetics, Spermatids metabolism, Spermatids ultrastructure, Lamin Type B metabolism, Nuclear Lamina metabolism, RNA, Messenger metabolism, Spermatogenesis genetics
- Abstract
The nuclear lamina (NL) is a filamentous protein meshwork, composed essentially of lamins, situated between the inner nuclear membrane and the chromatin. There is mounting evidence that the NL plays a role in spermatid differentiation during spermiogenesis. The mouse spermatid NL is composed of the ubiquitous lamin B1 and the spermatid-specific lamin B3, an N-terminally truncated isoform of lamin B2. However, nothing is known about the NL in human spermatids. We therefore investigated the expression pattern and localization of A-type lamins (A, C and C2) and B-type lamins (B1, B2 and B3) during human spermiogenesis. Here, we show that a lamin B3 transcript is present in human spermatids and that B-type lamins are the only lamins detectable in human spermatids. We determine that, as shown for their mouse counterparts, human lamin B3, but not lamin B2, induces strong nuclear deformation, when ectopically expressed in HeLa cells. Co-immunofluorescence revealed that, in human spermatids, B-type lamins are present at the nuclear periphery, except in the region covered by the acrosome, and that as the spermatid matures the B-type lamins recede towards the posterior pole. Only lamin B1 remains detectable on 33-47% of ejaculated spermatozoa. On spermatozoa selected for normal head density, however, this fell to <6%, suggesting that loss of the NL signal may be linked to complete sperm nucleus compaction. The similarities revealed between lamin expression during human and rodent spermiogenesis, strengthen evidence that the NL and lamin B3 have conserved functions during the intense remodelling of the mammalian spermatid nucleus., (© The Author 2014. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)
- Published
- 2015
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7. Prognosis factors of pregnancy after intrauterine insemination with the husband's sperm: conclusions of an analysis of 2,019 cycles.
- Author
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Dinelli L, Courbière B, Achard V, Jouve E, Deveze C, Gnisci A, Grillo JM, and Paulmyer-Lacroix O
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- Adult, Age Distribution, Comorbidity, Female, France epidemiology, Humans, Middle Aged, Pregnancy, Prevalence, Prognosis, Retrospective Studies, Risk Factors, Treatment Outcome, Fertilization in Vitro statistics & numerical data, Infertility, Female epidemiology, Infertility, Female therapy, Insemination, Artificial, Homologous statistics & numerical data, Pregnancy Outcome epidemiology, Pregnancy Rate
- Abstract
Objective: To identify the prognostic factors for pregnancy after intrauterine insemination with the husband's sperm (IUI-H)., Design: Retrospective study., Setting: A single university medical center., Patient(s): 851 couples, for 2,019 IUI-H cycles., Intervention(s): After controlled ovarian stimulation, IUI-H performed 36 hours after ovulation triggering or 24 hours after a spontaneous luteinizing hormone (LH) surge., Main Outcome Measure(s): Clinical pregnancy rate per cycle (PR) and delivery rate per cycle (DR)., Result(s): The overall PR was 14.8% and DR 10.8%. Higher PR and DR were observed for patients presenting with ovulation disorders (particularly polycystic ovary syndrome) or with male infertility. Secondary infertility in the woman appeared to be a positive prognostic factor as did a basal follicle-stimulating hormone (FSH) level ≤ 7 IU/L and ovulation triggering over spontaneous LH rise. The other parameters influencing the results were the women's age, the number of mature follicles obtained (≥ 2), the endometrial thickness (10-11 mm), and the number of progressive motile spermatozoa inseminated (>1 million)., Conclusion(s): In women aged ≤ 38 years, IUI-H should be considered as an option, particularly in cases of female infertility from ovulation disorders, in cases of a normal ovarian reserve, in cases of secondary infertility, or when ≥ 1 million progressive sperm are inseminated. Bifollicular stimulation is required. In other cases, in vitro fertilization should be discussed as the first-line treatment., (Copyright © 2014 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.)
- Published
- 2014
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8. Globozoospermia is mainly due to DPY19L2 deletion via non-allelic homologous recombination involving two recombination hotspots.
- Author
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Elinati E, Kuentz P, Redin C, Jaber S, Vanden Meerschaut F, Makarian J, Koscinski I, Nasr-Esfahani MH, Demirol A, Gurgan T, Louanjli N, Iqbal N, Bisharah M, Pigeon FC, Gourabi H, De Briel D, Brugnon F, Gitlin SA, Grillo JM, Ghaedi K, Deemeh MR, Tanhaei S, Modarres P, Heindryckx B, Benkhalifa M, Nikiforaki D, Oehninger SC, De Sutter P, Muller J, and Viville S
- Subjects
- Homozygote, Humans, Linkage Disequilibrium, Male, Point Mutation, Repetitive Sequences, Nucleic Acid, Gene Deletion, Homologous Recombination, Infertility, Male genetics, Membrane Proteins genetics
- Abstract
To date, mutations in two genes, SPATA16 and DPY19L2, have been identified as responsible for a severe teratozoospermia, namely globozoospermia. The two initial descriptions of the DPY19L2 deletion lead to a very different rate of occurrence of this mutation among globospermic patients. In order to better estimate the contribution of DPY19L2 in globozoospermia, we screened a larger cohort including 64 globozoospermic patients. Twenty of the new patients were homozygous for the DPY19L2 deletion, and 7 were compound heterozygous for both this deletion and a point mutation. We also identified four additional mutated patients. The final mutation load in our cohort is 66.7% (36 out of 54). Out of 36 mutated patients, 69.4% are homozygous deleted, 19.4% heterozygous composite and 11.1% showed a homozygous point mutation. The mechanism underlying the deletion is a non-allelic homologous recombination (NAHR) between the flanking low-copy repeats. Here, we characterized a total of nine breakpoints for the DPY19L2 NAHR-driven deletion that clustered in two recombination hotspots, both containing direct repeat elements (AluSq2 in hotspot 1, THE1B in hotspot 2). Globozoospermia can be considered as a new genomic disorder. This study confirms that DPY19L2 is the major gene responsible for globozoospermia and enlarges the spectrum of possible mutations in the gene. This is a major finding and should contribute to the development of an efficient molecular diagnosis strategy for globozoospermia.
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- 2012
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9. Sperm cryopreservation before cancer treatment: a 15-year monocentric experience.
- Author
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Bizet P, Saias-Magnan J, Jouve E, Grillo JM, Karsenty G, Metzler-Guillemain C, and Perrin J
- Subjects
- Adolescent, Adult, Hodgkin Disease complications, Hodgkin Disease therapy, Humans, Infertility, Male etiology, Male, Neoplasms complications, Retrospective Studies, Semen Analysis, Testicular Neoplasms complications, Testicular Neoplasms therapy, Cryopreservation methods, Fertility Preservation statistics & numerical data, Neoplasms therapy, Spermatozoa
- Abstract
Sperm banking is an important procedure to preserve fertility before cancer therapy. The aim of this study was to comprehensively analyse cryopreservation activity retrospectively for 1080 patients referred to the sperm bank for sperm cryopreservation before cancer treatment. This study included 1007 patients diagnosed with testicular cancer (TC) (41.7%), lymphoma (26%), other haematological cancers (9.4%) or other types of cancer (22.8%); of these, 29 patients did not produce any semen sample and cryopreservation was impossible for 67 patients. Semen characteristics before treatment were within normal ranges, except moderate asthenospermia. Sperm concentration was significantly lower in TC than in non-TC. Straws from 57 patients (6.3%) were used in assisted reproductive technologies, which led to a 46.8% cumulative birth rate. Straws were destroyed for 170 patients (18.7%) and 140 patients performed semen analyses after cancer therapy. After an average delay of 22.5 months after the end of therapy, 43 patients (30.7%) exhibited azoospermia. This study of a large population of cancer patients revealed a high level of successful sperm storage. Utilization of cryopreserved spermatozoa led to good chances of fatherhood. Nevertheless, sperm banks should be aware of the low rates of straw use and straw destruction by cancer patients., (Copyright © 2011 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.)
- Published
- 2012
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10. [Do couples who procreate through sperm donation inform their children?].
- Author
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Araya R, Rouzier J, Perrin J, Carez S, Saias-Magnan J, Noizet A, Grillo JM, and Metzler-Guillemain C
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- Child, Child of Impaired Parents psychology, Child of Impaired Parents statistics & numerical data, Data Collection, Female, Humans, Infertility, Male, Male, Pregnancy, Social Environment, Surveys and Questionnaires, Family Characteristics, Parent-Child Relations, Spermatozoa, Tissue and Organ Procurement ethics, Truth Disclosure ethics
- Abstract
Objectives: An amendment to the French bioethics law allowing children conceived by gamete donation to know the identity of donors is proposed, while no study can assess the proportion of parents in France that disclose the nature of conception to their donor conceived offspring. The aim of our study was to know whether couples who wish to inform their offspring actually did it., Patients and Methods: We sent a questionnaire to parents who had expressed an intention to disclose the nature of conception to their future offspring conceived by sperm donation. This allowed us to evaluate the number of couples who inform their offspring, and the couple and offspring feelings when information was given., Results: Among 38 questionnaires sent, 20 couples answered. Fourteen informed their offspring about the nature of conception, most having lived serenely this moment. 47% of offspring have reacted with indifference. While 19 couples informed their friends or family, six couples did not inform their offspring, and two of them no longer want to disclose anymore., Conclusion: Careful thought before the beginning of assisted reproductive technology and support after birth are needed to help couples communicate information to their offspring. Without this communication, any policy of openness to know donor related data seems vain., (Copyright © 2011 Elsevier Masson SAS. All rights reserved.)
- Published
- 2011
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11. Using statistical and machine learning to help institutions detect suspicious access to electronic health records.
- Author
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Boxwala AA, Kim J, Grillo JM, and Ohno-Machado L
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- Humans, Logistic Models, Pilot Projects, Sensitivity and Specificity, Software Validation, United States, Artificial Intelligence, Computer Security, Electronic Health Records, Management Audit methods
- Abstract
Objective: To determine whether statistical and machine-learning methods, when applied to electronic health record (EHR) access data, could help identify suspicious (ie, potentially inappropriate) access to EHRs., Methods: From EHR access logs and other organizational data collected over a 2-month period, the authors extracted 26 features likely to be useful in detecting suspicious accesses. Selected events were marked as either suspicious or appropriate by privacy officers, and served as the gold standard set for model evaluation. The authors trained logistic regression (LR) and support vector machine (SVM) models on 10-fold cross-validation sets of 1291 labeled events. The authors evaluated the sensitivity of final models on an external set of 58 events that were identified as truly inappropriate and investigated independently from this study using standard operating procedures., Results: The area under the receiver operating characteristic curve of the models on the whole data set of 1291 events was 0.91 for LR, and 0.95 for SVM. The sensitivity of the baseline model on this set was 0.8. When the final models were evaluated on the set of 58 investigated events, all of which were determined as truly inappropriate, the sensitivity was 0 for the baseline method, 0.76 for LR, and 0.79 for SVM., Limitations: The LR and SVM models may not generalize because of interinstitutional differences in organizational structures, applications, and workflows. Nevertheless, our approach for constructing the models using statistical and machine-learning techniques can be generalized. An important limitation is the relatively small sample used for the training set due to the effort required for its construction., Conclusion: The results suggest that statistical and machine-learning methods can play an important role in helping privacy officers detect suspicious accesses to EHRs.
- Published
- 2011
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12. Tobacco consumption and benzo(a)pyrene-diol-epoxide-DNA adducts in spermatozoa: in smokers, swim-up procedure selects spermatozoa with decreased DNA damage.
- Author
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Perrin J, Tassistro V, Mandon M, Grillo JM, Botta A, and Sari-Minodier I
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- Adult, Benzo(a)pyrene metabolism, Benzopyrans adverse effects, Case-Control Studies, Cell Separation methods, Cells, Cultured, DNA Damage physiology, Epoxy Compounds adverse effects, Humans, Male, Semen Analysis, Tobacco Products, Benzopyrans metabolism, DNA Adducts metabolism, Epoxy Compounds metabolism, Smoking metabolism, Sperm Retrieval, Spermatozoa metabolism, Spermatozoa pathology
- Abstract
Objective: To analyze the distribution of benzo(a)pyrene-diol-epoxide (BPDE)-DNA adducts in spermatozoa selected and nonselected by a swim-up procedure with relation to smoking habits., Design: Comparative study., Setting: Public university and public university hospital., Patient(s): Seventy-nine men (37 smokers and 42 nonsmokers) who visited an infertility clinic for diagnostic., Intervention(s): Tobacco and environmental exposure assessment, semen sample analysis, swim-up procedure, BPDE-DNA adduct immunolabeling., Main Outcome Measure(s): BPDE-DNA adduct quantification in selected (SEL-SPZ) and nonselected (NONSEL-SPZ) spermatozoa. Data were normalized by using a normalized fluorescence value (NFV)., Result(s): The mean NFV (±SD) in SEL-SPZ was significantly higher in smokers than in nonsmokers (18.9±11.5 vs. 10.5±10.4, respectively). Within smokers, a paired analysis (SEL-SPZ and NONSEL-SPZ) showed that NFV was significantly lower in SEL-SPZ than in NONSEL-SPZ (20.0±11.3 vs. 31.5±16.0, respectively). Conversely, within nonsmokers, the mean NFV was higher in SEL-SPZ than in NONSEL-SPZ (10.3±10.6 vs 4.3±7.1, respectively)., Conclusion(s): Tobacco consumption is associated with BPDE-DNA adducts in spermatozoa. In smokers, semen processing by swim-up recovers potentially fertilizing spermatozoa that show a significantly lower amount of BPDE-DNA adducts compared with NONSEL-SPZ. Further study is needed to improve the spermatozoa selection in smoking patients requiring assisted reproductive technologies., (Copyright © 2011 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.)
- Published
- 2011
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13. [Initial results of a novel technique for sperm retrieval in male infertility due to refractory retrograde ejaculation].
- Author
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Perrin J, Saïas-Magnan J, Lanteaume A, Thiry-Escudié I, Serment G, Bladou F, Gamerre M, Grillo JM, and Karsenty G
- Subjects
- Adult, Case-Control Studies, Humans, Male, Prospective Studies, Ejaculation, Infertility, Male etiology, Sexual Dysfunction, Physiological complications, Sperm Retrieval
- Abstract
Aims: To describe and evaluate a novel technique of spermatozoa retrieval from patients suffering from infertility secondary to refractory retrograde ejaculation., Method: Prospective study to compare mobility and vitality of spermatozoa obtained from urine (U) after oral modification of chemical parameter (PH, Osmolarity) versus from endovesical instillation of sterile spermatozoa culture medium before ejaculation (F). Patients were their own controls. Twelve month prospective follow-up was achieved to document the results of assisted procreation., Results: Eight patients were included and mobility and vitality were improved in all patients after F technique was compared to U technique. With U technique, eight patients on eight had negative defrost test; after F technique, seven patients on eight had a positive defrost test and could therefore have access to assisted reproduction techniques. Four couples had five ICSI and obtained three pregnancies leading to five births., Conclusion: Endovesical instillation of sterile spermatozoa culture medium before ejaculation was a safe and effective technique to improve spermatozoa quality in male infertility related to refractory retrograde ejaculation., (Copyright © 2010. Published by Elsevier Masson SAS.)
- Published
- 2011
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14. Anomaly and signature filtering improve classifier performance for detection of suspicious access to EHRs.
- Author
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Kim J, Grillo JM, Boxwala AA, Jiang X, Mandelbaum RB, Patel BA, Mikels D, Vinterbo SA, and Ohno-Machado L
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- Humans, Logistic Models, Privacy, Sensitivity and Specificity, Artificial Intelligence, Computer Security, Electronic Health Records
- Abstract
Our objective is to facilitate semi-automated detection of suspicious access to EHRs. Previously we have shown that a machine learning method can play a role in identifying potentially inappropriate access to EHRs. However, the problem of sampling informative instances to build a classifier still remained. We developed an integrated filtering method leveraging both anomaly detection based on symbolic clustering and signature detection, a rule-based technique. We applied the integrated filtering to 25.5 million access records in an intervention arm, and compared this with 8.6 million access records in a control arm where no filtering was applied. On the training set with cross-validation, the AUC was 0.960 in the control arm and 0.998 in the intervention arm. The difference in false negative rates on the independent test set was significant, P=1.6×10(-6). Our study suggests that utilization of integrated filtering strategies to facilitate the construction of classifiers can be helpful.
- Published
- 2011
15. [The spinal cord injured patient: semen quality and management by Assisted Reproductive Technology].
- Author
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Perrin J, Saïas-Magnan J, Thiry-Escudié I, Gamerre M, Serment G, Grillo JM, Guillemain C, and Karsenty G
- Subjects
- Ejaculation, Female, Humans, Infertility, Male physiopathology, Infertility, Male therapy, Insemination, Artificial, Homologous, Male, Pregnancy, Semen Analysis, Sperm Retrieval, Infertility, Male etiology, Pregnancy Rate, Reproductive Techniques, Assisted, Spinal Cord Injuries complications
- Abstract
Men with spinal cord injury present a unique infertile population. Only 10 % of them can father children without medical assistance, owing to potential impairments in erection, ejaculation and semen quality. The algorithm typically followed is to retrieve semen by Penile Vibratory Stimulation, in case of failure by Electro Ejaculation. Most of these patients have normal sperm concentrations but abnormally low sperm motility and vitality in the ejaculate. The reasons for poor semen quality in spinal cord injured men are reviewed. If semen cannot be obtained by Electro Ejaculation, or if the ejaculate from Penile Vibratory Stimulation or Electro Ejaculation contains an insufficient quantity or quality of sperm for in vitro fertilization with intracytoplasmic sperm injection, then retrieval of sperm from reproductive tissues is attempted. Despite abnormal semen quality, successful pregnancies with sperm from spinal cord injured male partners have occurred by intravaginal insemination, intrauterine insemination, and in vitro fertilization with intracytoplasmic sperm injection. The prevailing pregnancy and fecundity rates in couples with a spinal cord injured male partner are reviewed., (Copyright © 2010 Elsevier Masson SAS. All rights reserved.)
- Published
- 2010
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16. Ultrasound monitoring in patients undergoing in-vitro fertilization after methotrexate treatment for ectopic pregnancy.
- Author
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Provansal M, Agostini A, Lacroix O, Gerbeau S, Grillo JM, and Gamerre M
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- Adult, Chorionic Gonadotropin therapeutic use, Female, Humans, Ovarian Follicle drug effects, Pregnancy, Pregnancy Tests, Pregnancy, Ectopic diagnostic imaging, Prognosis, Retrospective Studies, Treatment Outcome, Ultrasonography, Abortifacient Agents, Nonsteroidal therapeutic use, Endometrium diagnostic imaging, Endometrium drug effects, Fertilization in Vitro adverse effects, Methotrexate therapeutic use, Ovarian Follicle diagnostic imaging, Pregnancy, Ectopic drug therapy
- Abstract
Objectives: To compare sonographic characteristics of the endometrium and follicles during in-vitro fertilization (IVF) before and after methotrexate (MTX) treatment for ectopic pregnancy., Methods: This retrospective study, conducted at Conception Hospital from January 2000 to July 2007, included all patients diagnosed with an ectopic pregnancy resulting from IVF treatment that was treated with MTX and who then underwent another IVF cycle. We compared the number and size of follicles and the endometrial thickness and quality on the day of human chorionic gonadotropin injection in the cycles before and after the MTX treatment to determine whether MTX had any effect., Results: Eleven patients were included in the study. The median interval between the IVF cycle resulting in ectopic pregnancy and the first IVF cycle after MTX therapy was 180 (range, 150-900) days. There was no statistically significant difference between the before and after MTX treatment groups with respect to number of follicles (14 (3-20) vs. 9 (4-16), P = 0.12), follicle size (16.5 (14.7-21.7) mm vs. 17.8 (14.9-19.8) mm, P = 0.37), endometrial thickness (10.0 (9.5-12.0) mm vs. 10.0 (7.5-14.0) mm, P = 0.31) or endometrial quality (P = 0.32). Four women became pregnant during the IVF cycle following MTX treatment., Conclusions: Ultrasound monitoring showed no modification of the characteristics of the endometrium or follicles during IVF after MTX treatment for ectopic pregnancy., (Copyright 2009 ISUOG. Published by John Wiley & Sons, Ltd.)
- Published
- 2009
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17. Occupational exposures obtained by questionnaire in clinical practice and their association with semen quality.
- Author
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De Fleurian G, Perrin J, Ecochard R, Dantony E, Lanteaume A, Achard V, Grillo JM, Guichaoua MR, Botta A, and Sari-Minodier I
- Subjects
- Adult, Electromagnetic Fields adverse effects, Hot Temperature adverse effects, Humans, Infertility, Male etiology, Male, Metals, Heavy adverse effects, Middle Aged, Occupational Diseases complications, Pesticides adverse effects, Solvents adverse effects, Vibration adverse effects, Air Pollutants, Occupational adverse effects, Occupational Exposure adverse effects, Semen drug effects, Semen Analysis
- Abstract
In industrial countries, evidence suggests that semen quality has been steadily decreasing over the past 5 decades. We employed a short questionnaire to examine the association between self-reported physical or chemical occupational exposures and semen quality. The study included 402 men consulting for couple infertility (314 with oligospermia, asthenospermia, or teratospermia and 88 with normal semen; World Health Organization criteria). Exposure effects on global sperm quality and total sperm count, sperm motility, and sperm morphology were investigated. We found significant associations between semen impairment and occupational risk factors such as exposure to heavy metals (adjusted odds ratio [OR] = 5.4; 95% confidence interval [CI], 1.6-18.1), solvents (OR = 2.5; 95% CI, 1.4-4.4), fumes (OR = 1.9; 95% CI, 1.1-3.4), and polycyclic aromatic hydrocarbons (OR = 1.9; 95% CI, 1.1-3.5). Exposure to pesticides or cement was nearly significant (OR = 3.6; 95% CI, 0.8-15.8, and OR = 2.5; 95% CI, 0.95-6.5, respectively). Physical risk factors were associated with some sperm anomalies, such as mechanical vibrations with oligospermia and teratospermia as well as excess heat and extended sitting periods with impaired motility. Exposure to ionizing radiation and electromagnetic fields was not associated with semen impairment; these results, however, may be skewed, because very few subjects reported such exposure. Despite the small dataset, self-reported exposures were correlated with semen impairment. This approach may be recommended in routine clinical practice to seek relationships between occupational exposures to reprotoxic agents and impaired semen parameters. This knowledge would allow preventive measures in the workplace to be established and could be complemented by the use of biomarkers to better characterize exposure to chemical substances and their spermiotoxic effects.
- Published
- 2009
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18. [What are at present the real hopes of pregnancy after ovarian cryopreservation?].
- Author
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Courbière B, Provansal M, Saias-Magnan J, Guillemain C, Noizet A, Grillo JM, and Gamerre M
- Subjects
- Antineoplastic Agents adverse effects, Female, Fertilization in Vitro, Humans, Infertility, Female etiology, Pregnancy, Primary Ovarian Insufficiency chemically induced, Radiation Injuries, Cryopreservation, Infertility, Female therapy, Organ Preservation, Ovarian Follicle physiology, Ovary transplantation, Tissue Preservation methods
- Abstract
Survival rates for cancers that occur in childhood and adolescence have improved over the last decades, and preservation of future fertility in these patients has become a relevant issue. Premature ovarian failure is a consequence of exposing women to chemotherapeutic drugs and ionizing radiation. Ovarian cryopreservation is an alternative to cryopreservation of embryos or oocytes for theses patients. Ovarian cryopreservation aims to reimplant ovarian tissue after complete remission into the pelvic cavity (orthotopique site) or a heterotopic site like the abdominal wall or the forearm. In vitro folliculogenesis, that aims at the maturation of ovarian cortex primordial follicles cryopreserved for a FIV, is still in an experimental research stage. In this review, the objective was to evaluate the real hopes of pregnancy after ovarian cryopreservation. Indeed, many teams offer ovarian cryopreservation at present time, although only two pregnancies have been achieved to date. In both cases, it can be discussed whether the fertilized oocyte originated from the transplant or from the native ovary. Furthermore, the potential for reintroduction of cancerous cells may limit this technique in cancers that are known to have a risk of ovarian dissemination. The hopes engendered by ovarian cryopreservation, but also its limits, must be explained to the patients before an ovarian surgery for cryopreservation.
- Published
- 2007
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19. Evolution of DNA strand-breaks in cultured spermatocytes: the Comet Assay reveals differences in normal and gamma-irradiated germ cells.
- Author
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Perrin J, Lussato D, De Méo M, Durand P, Grillo JM, Guichaoua MR, Botta A, and Bergé-Lefranc JL
- Subjects
- Animals, Antimetabolites, Bromodeoxyuridine, Caspases metabolism, Cell Survival radiation effects, Cells, Cultured, Coculture Techniques, Comet Assay, Germ Cells ultrastructure, Kinetics, Male, Rats, Rats, Wistar, Sertoli Cells radiation effects, Sertoli Cells ultrastructure, Spermatocytes ultrastructure, DNA Damage radiation effects, Gamma Rays, Germ Cells radiation effects, Spermatocytes radiation effects
- Abstract
In reproductive toxicity assessment, in vitro systems can be used to determine mechanisms of action of toxicants. However, they generally investigate the immediate effects of toxicants, on isolated germ cells or spermatozoa. We report here the usefulness of in vitro cultures of rat spermatocytes and Sertoli cells, in conjunction with the Comet Assay to analyze the evolution of DNA strand-breaks and thus to determine DNA damage in germ cells. We compared cultures of normal and gamma-irradiated germ cells. In non-irradiated spermatocytes, the Comet Assay revealed the presence of DNA strand-breaks, which numbers decreased with the duration of the culture, suggesting the involvement of DNA repair mechanisms related to the meiotic recombination. In irradiated cells, the evolution of DNA strand-breaks was strongly modified. Thus our model is able to detect genotoxic lesions and/or DNA repair impairment in cultured spermatocytes. We propose this model as an in vitro tool for the study of genotoxic injuries on spermatocytes.
- Published
- 2007
- Full Text
- View/download PDF
20. [Ovarian cryopreservation: clinical and ethical problems arisen further to study on cases of ovarian cryopreservation].
- Author
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Courbière B, Porcu G, Saias-Magnan J, Guillemain C, Noizet A, Michel G, Grillo JM, and Gamerre M
- Subjects
- Adolescent, Adult, Child, Child, Preschool, Female, Humans, Infertility, Female etiology, Infertility, Female surgery, Leukemia therapy, Neoplasms therapy, Ovary transplantation, Reproductive Techniques, Assisted, Retrospective Studies, Cryopreservation ethics, Ovary physiology
- Abstract
Objective: An early ovarian failure and sterility, on patients often still young, may result from some of the anticancerous treatments. Ovarian cryopreservation is a research way for fertility restoration on patients who will have gonadotoxic treatment. The aim of our work was to study the cases of ovarian tissue preserved in our Department, enabling us to assess the clinical and ethical problems of this technique., Patients and Methods: A retrospective study was conducted from January 2001 onwards up to October 2005, on 14 patients, whose ovarian cortex was frozen our the University Fertility Center (Hôpital de la Conception, Marseille, France)., Results: In our study, the patients' average age was 17+/-5.5 and the median was 14 years [13-24]. Eight patients were under 18 (11+/-3-years-old). The cryopreservation indications were especially haematological (N = 9). More than half of the patients (N = 8) had undergone a gonadotoxic treatment before ovarian cryopreservation., Discussion and Conclusion: Ovarian cryopreservation is still a recent technology in reproductive medicine. The future of ovarian tissue after thawing, with its different techniques of autograft, is still not consensual. Experimental research remains essential to improve the freezing protocols and ovarian transplant in human medicine.
- Published
- 2006
- Full Text
- View/download PDF
21. Meiotic arrest at the midpachytene stage in a patient with complete azoospermia factor b deletion of the Y chromosome.
- Author
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Perrin J, Metzler-Guillemain C, Karsenty G, Grillo JM, Mitchell MJ, and Guichaoua MR
- Subjects
- Adult, Genetic Loci, Humans, Immunohistochemistry, Male, Spermatozoa pathology, Chromosomes, Human, Y, Gene Deletion, Meiosis, Oligospermia genetics, Oligospermia pathology, Seminal Plasma Proteins genetics, Sex Chromosome Aberrations
- Abstract
Objective: To study the meiosis of a patient with complete azoospermia factor (AZF)b deletion of the Y chromosome., Design: Case report., Setting: La Conception University Hospital, Marseille, France., Patient(s): One azoospermic patient., Intervention(s): Yq deletion testing, testicular sperm extraction, and meiotic study with immunocytochemistry., Main Outcome Measure(s): Abnormal synapsis rates in spermatocytes., Result(s): We found that most spermatocytes were at an early stage of meiosis. Half of the meiotic germ cells analyzed showed asynapsis, which was mostly extended or total. Discontinuity in the synaptonemal complex was seen in one third of the meiotic cells analyzed. An unusually small number of normal pachytene nuclei were found, all at early pachytene substages., Conclusion(s): This is the first demonstration that the synaptic process is impaired in a man with complete deletion of the AZFb interval. Our findings provide evidence that the pachytene checkpoint is situated at the midpachytene substage in humans.
- Published
- 2006
- Full Text
- View/download PDF
22. [Optimization of artificial inseminations with donor semen: a four-year experience].
- Author
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Achard V, Perrin J, Saias-Magnan J, Noizet A, Grillo JM, and Paulmyer-Lacroix O
- Subjects
- Adult, Female, Humans, Infertility, Male therapy, Male, Pregnancy, Sperm Count, Sperm Motility, Treatment Outcome, Insemination, Artificial, Heterologous methods
- Abstract
Objective: Our aim was to analyse the results of a donor insemination program using ovarian stimulation, swim-up sperm preparation and intrauterine insemination proposed to women with a maximum age of 39. Incidence of several clinical and biological parameters on success rates was investigated., Patients and Methods: Retrospective analysis of the results of 249 cycles performed in 106 couples during a four-year period is reported., Results: Overall pregnancy rate of 28.1% and delivery rate of 22% per cycle were achieved, with a multiple pregnancy rate of 11.4%. Most of the pregnancies (84%) were obtained before the fourth insemination. Among the different parameters studied the total number of motile sperm inseminated was found to be the most important factor for success rate: pregnancy rate per cycle reached 40.4% if more than 1.5 million progressive sperm were inseminated vs. 24.7% if they were less than 1.5 million (P<0.05)., Discussion and Conclusion: In precise conditions, outcome of inseminations with donor semen can reach satisfying pregnancy rates, being a valuable help for couples suffering of long-time infertility.
- Published
- 2005
- Full Text
- View/download PDF
23. Meiotic anomalies in infertile men with severe spermatogenic defects.
- Author
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Guichaoua MR, Perrin J, Metzler-Guillemain C, Saias-Magnan J, Giorgi R, and Grillo JM
- Subjects
- Adult, Azure Stains, Biopsy, Case-Control Studies, Cell Nucleus pathology, Coloring Agents, Humans, Male, Microscopy, Fluorescence, Middle Aged, Retrospective Studies, Spermatocytes pathology, Testis pathology, Meiosis, Oligospermia pathology, Spermatogenesis
- Abstract
Background: This study was aimed at evaluating the rate of pairing failure in pachytene spermatocytes of patients presenting either an obstructive (O) or a non-obstructive (NO) infertility., Methods: Forty-one patients and 13 controls underwent testicular biopsy. Among the patients, 19 had an O infertility and 22 a NO infertility. Preparations of all patients and controls were Giemsa-stained, and synaptonemal complexes from nine of these patients and one control were immunostained., Results: In all, 2931 pachytene nuclei were analysed. The mean rate of asynapsed nuclei from the NO group (25.4%) was significantly higher than that of the O group (9.8%). There was no significant difference between the O group and the controls (10.6%). Immunocytochemistry showed that the number of pachytene nuclei decreased from the early to late pachytene sub-stage in all patients. Two NO patients, one azoospermic and one oligozoospermic, had a high percentage of asynapsed nuclei (86 and 91.8% respectively); one of these patients also presented a precocious localized separation of sister chromatids., Conclusion: high levels of extended asynapsis could arise from a primary meiotic defect which may be responsible for 9% of the NO male infertilities at our centre. The prevalence of early pachytene substages suggests that the pachytene checkpoint is localized at the mid-pachytene stage in humans.
- Published
- 2005
- Full Text
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24. A truncated HCV core protein elicits a potent immune response with a strong participation of cellular immunity components in mice.
- Author
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Alvarez-Obregón JC, Dueñas-Carrera S, Valenzuela C, and Grillo JM
- Subjects
- Animals, Antibody Specificity, Female, Gene Expression, Hepacivirus genetics, Hepatitis C Antibodies biosynthesis, Hepatitis C Antigens chemistry, Hepatitis C Antigens genetics, Immunity, Cellular, Immunoglobulin G biosynthesis, In Vitro Techniques, Interferon-gamma genetics, Lymphocyte Activation, Mice, Mice, Inbred BALB C, Mice, Inbred C57BL, Peptide Fragments chemistry, Peptide Fragments genetics, Peptide Fragments immunology, Th1 Cells immunology, Viral Core Proteins chemistry, Viral Core Proteins genetics, Hepacivirus immunology, Hepatitis C Antigens immunology, Viral Core Proteins immunology
- Abstract
The immunogenicity of a truncated HCV core protein (Co.120) was studied in BALB/c and C57BL/6 mice, given three intramuscular injections of antigen, adjuvanted with either aluminum hydroxide or Freund's adjuvant. A rapid antibody response was noted after the first dose, with both strains of mice eventually exhibiting comparable levels of anti-core IgG (titers >1:100000), with a mixed IgG1/IgG2a subclass response. Spleen cells from Co.120-immunized mice gave a significant specific proliferative response. IFN-gamma gene expression was also detected after an ex-vivo specific stimulation of spleen cells in all immunized mice. This response was independent of dose, H-2 genetic background or type of adjuvant. The results indicated that immunization with the Co.120 protein elicits a potent anti-HCV humoral and cellular immune response.
- Published
- 2001
- Full Text
- View/download PDF
25. [Pregnancy outcome and the status of the newborn infant after ICSI at La Conception Hospital in Marseille].
- Author
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Porcu-Buisson G, Capelle M, Paulmyer-Lacroix O, Saïas-Magnan J, Carles F, Noizet A, Grillo JM, and Gamerre M
- Subjects
- Congenital Abnormalities epidemiology, Embryo Transfer, Female, France epidemiology, Gestational Age, Humans, Infant, Premature, Male, Pregnancy, Pregnancy, Multiple, Prospective Studies, Twins, Infant, Newborn, Pregnancy Outcome, Sperm Injections, Intracytoplasmic
- Abstract
Objective: To evaluate obstetric outcome of pregnancies and pediatric follow-up of children born after ICSI procedure., Material and Method: Prospective study from october 1994 to September 1998 in medical assisted procreations center in La Conception hospital in Marseilles. Three hundred forty-two couples undergoing ICSI procedures., Interventions: analysis of pregnancy rates, prematurity rates, obstetric outcome and frequency of congenital malformations., Results: One hundred seventy-eight pregnancies have been obtained with 111 children. Multiple pregnancy rate arise 29.7%. The average term at birth in the singleton pregnancies (38.7 weeks) is higher than in the twins (35.7 weeks). The prematurity rate of delivery before 35 weeks of gestation is about 9.6%. Seven of 111 neonates was born with a congenital malformation, no cardiovascular one. One of boys presents a bilateral cryptorchidism with severe bilateral hypotrophy., Conclusions: Our results are similar with those of others teams. Congenital malformation rates is near rates reported in others studies. However, no bilateral cryptorchidism with bilateral severe hypotrophy has been yet reported in literature.
- Published
- 2001
- Full Text
- View/download PDF
26. [Morphological aspect of embryos obtained after fertilization in vitro for male factor or ICSI].
- Author
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Saias-Magnan J, Mendizabal H, Guichaoua MR, Carles F, Grillo JM, and Luciani JM
- Subjects
- Adult, Embryo Implantation, Embryo Transfer, Embryo, Mammalian physiology, Female, Humans, Male, Pregnancy, Embryo, Mammalian anatomy & histology, Fertilization in Vitro, Infertility, Male therapy, Sperm Injections, Intracytoplasmic
- Abstract
Objective: In order to study the eventual impact on fertilization and embryo characteristics of the microinjection procedure we compared the quality of the embryos obtained by ICSI with those of in vitro fertilization with male factors (MF IVF)., Material and Methods: One hundred thirty-four cycles of IVF treatment (group 1) were selected with oligoasthenozoospermia according to WHO criteria with a total number of motile spermatozoa between 500,000 and 1 million. One thousand eighty-eight mature oocytes and 486 embryos were obtained. One hundred forty-three cycles of intracytoplasmic sperm injection (group 2) were performed in couples whose in vitro fertilization was imparticable because of extreme sperm impairment. One thousand one hundred forty-seven mature oocytes were injected and 626 embryos were obtained., Results: In group 1, the pregnancy rate per embryo transfer and the implantation rate were respectively 22.7% and 12.3%. In group 2, the pregnancy rate per embryo transfer was 37.1% and the implantation rate was 17%. The statistical analysis of the embryos obtained in the two different groups did not demonstrate any difference in the distribution of the more regular and less fragmented embryos (group A) and those of the more irregular and fragmented embryos (group B). No statistical difference was demonstrated in the chronology of the division of these embryos (groups 1 and 2)., Conclusion: The pregnancy rate by cycle and by transfer reported by ICSI (p < 0.003 and p < 0.015 respectively) could be related to a significantly higher mean number of transferred embryos (2.65 vs 2.02) in probable relation with a higher cleavage rate (p < 0.00001).
- Published
- 2000
- Full Text
- View/download PDF
27. Failure of pregnancy after intracytoplasmic sperm injection with decapitated spermatozoa: case report.
- Author
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Saïas-Magnan J, Metzler-Guillemain C, Mercier G, Carles-Marcorelles F, Grillo JM, and Guichaoua MR
- Subjects
- Adult, Female, Humans, Male, Pregnancy, Pregnancy Outcome, Spermatozoa pathology, Infertility, Male, Insemination, Artificial, Sperm Head pathology
- Abstract
The case of a couple with a history of long standing primary infertility is reported in which the man presented with a decapitated sperm defect. The woman had a normal history and presented with normal clinical characteristics. The couple underwent one unsuccessful conventional in-vitro fertilization (IVF). Subsequently, embryos were obtained and transferred after assisted fertilization attempts: in all, three subzonal inseminations and four intracytoplasmic sperm injections. A total of 49 mature oocytes was injected in both studies, 25 embryos obtained and 20 embryos transferred, three of them after freezing and thawing. Despite the good embryo morphology, implantation was unsuccessful and no pregnancy occurred. The failure of implantation may have resulted from an arrest in early embryonic development related to the sperm anomaly. One hypothesis is that transferred embryos may carry a chromosomal imbalance that prevents them from progressing to the blastocyst stage. Nevertheless, we cannot exclude the possibility that the woman is responsible for the implantation failure. Co-culture associated with a further attempt could provide information regarding the ability of embryos to progress to the blastocyst stage and implant.
- Published
- 1999
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28. Ultrastructural and immunocytochemical evidences of core-particle formation in the methylotrophic Pichia pastoris yeast when expressing HCV structural proteins (core-E1).
- Author
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Falcón V, García C, de la Rosa MC, Menéndez I, Seoane J, and Grillo JM
- Subjects
- Gene Expression, Humans, Microscopy, Immunoelectron, Viral Core Proteins genetics, Viral Envelope Proteins genetics, Virion ultrastructure, Hepacivirus physiology, Pichia ultrastructure, Viral Core Proteins biosynthesis, Viral Envelope Proteins biosynthesis, Virus Assembly
- Abstract
Particulate antigens of the Hepatitis C virus (HCV) are reported for the first time by transmission electron microscopy in Pichia pastoris. The yeast was cloned to express the first 339 NH2-terminal amino acids of the HCV polyprotein (C-E1.339 polypeptide). The C-E1.339 polypeptide covers the putative 191 aa of the core protein (aa 1-191) and 148 aa of the E1 envelope antigen (aa 192-339). Virus-like particles (VLP) with diameters ranging from 20 nm to 30 nm were specifically observed in those cells expressing the HCV polyprotein. The VLP appeared along the membrane of the endoplasmic reticulum, but were fundamentally localized in vacuoles, either free or inside autophagic bodies. Clustered particles, chains of particles, high-density reticular structures, and crystalloid bodies were also detected, the last one being an orderly arrangement of particles with 20 nm diameters. The crystal-associated particles are well differentiated from the intracellular VLP because of their uniform size and shape. We argue that membrane components are retained in the architecture of the VLP, conferring to this particle certain heterogeneity. Both kinds of particles, the VLP formed after treatment with NP-40 and the crystal-associated particles, were core protein-positives. Whether they reflect mature HCV nucleocapsid or intermediary states in the viral nucleocapsid morphogenesis remains unknown. We conclude that, like mammalian cell lines, the P. pastoris yeast could be an appropriate host for the analysis of HCV polyprotein processing and, eventually, virus assembly.
- Published
- 1999
- Full Text
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29. [Intrauterine insemination with the husband's sperm: conclusions of five years experience].
- Author
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Paulmyer-Lacroix O, Mollé L, Noizet A, Guérin A, Mollar M, Gamerre M, and Grillo JM
- Subjects
- Adult, Female, Humans, Infertility etiology, Male, Pregnancy, Retrospective Studies, Sperm Motility, Time Factors, Infertility therapy, Insemination, Artificial, Homologous methods, Pregnancy Outcome
- Abstract
We analyzed retrospectively 936 intrauterine insemination (IUI) cycles with husband's sperm (384 couples). Superovulation and induction monitoring occurred in the majority of cases; IUI was timed 36-40 h after ovulatory hCG. The overall pregnancy rate per cycle (PR/C) was 11.4% (107/936). Data analysis demonstrated that PR/C decreased with infertility duration, woman's age (especially after 38 years old) and number of attempts (significantly after the 4 th cycle). Superovulation (in particular by antiestrogens) and induction monitoring seemed to provide an increase in cycle pregnancy rate. Highest PR/C were observed in dysovulation and male infertility groups. In cases of sperm defects, our data showed that 2 parameters had a significant influence on pregnancy outcome: the number of motile progressive inseminated spermatozoa (> 300,000) and the spermatozoa survival rate after 24 h (> 50%). As a conclusion, intrauterine insemination can be proposed as a satisfying treatment of infertility, if precise protocol is followed and its indications well-defined.
- Published
- 1998
30. Invertase secretion in Hansenula polymorpha under the AOX1 promoter from Pichia pastoris.
- Author
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Rodriguez L, Narciandi RE, Roca H, Cremata J, Montesinos R, Rodriguez E, Grillo JM, Muzio V, Herrera LS, and Delgado JM
- Subjects
- Alcohol Oxidoreductases genetics, Cell Compartmentation, Culture Media, Gene Expression Regulation, Fungal, Genes, Fungal, Genetic Vectors, Glycerol pharmacology, Glycoside Hydrolases genetics, Methanol metabolism, Pichia drug effects, Pichia genetics, Pichia growth & development, Promoter Regions, Genetic, Recombinant Fusion Proteins metabolism, Saccharomyces cerevisiae enzymology, Saccharomyces cerevisiae genetics, beta-Fructofuranosidase, Glycoside Hydrolases metabolism, Pichia metabolism
- Abstract
A DNA fragment containing a transcription regulating region of the alcohol oxidase (AOX1) gene from the methylotrophic yeast Pichia pastoris was used in the construction of a vector for the expression of heterologous proteins in the methylotrophic yeast Hansenula polymorpha. We used this vector to clone the SUC2 gene from Saccharomyces cerevisiae into H. polymorpha yeast. The culture conditions for invertase production using a fed-batch culture were studied. More than 1.5 x 10(3) U/ml of biologically active invertase (1 g/l) were secreted to the cellular periplasmic space. The fermentative process was scaled up to 50 l. Invertase produced from H. polymorpha was glycosylated, but it contained significantly less carbohydrate than protein produced by S. cerevisiae. Using the Western-blot technique, it was observed that invertase secreted from H. polymorpha and invertase secreted from S. cerevisiae showed common antigenic determinants.
- Published
- 1996
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- View/download PDF
31. Influence of the morphological aspect of embryos obtained by in vitro fertilization on their implantation rate.
- Author
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Grillo JM, Gamerre M, Lacroix O, Noizet A, and Vitry G
- Subjects
- Embryo, Mammalian physiology, Female, Humans, Male, Statistics as Topic, Embryo Implantation physiology, Embryo, Mammalian anatomy & histology, Embryonic and Fetal Development physiology, Fertilization in Vitro
- Abstract
In a series of 500 transfers of embryos obtained by in vitro fertilization, we examined the implantation rate of 1356 embryos transferred in utero. The average implantation rate per embryo was 15.1% and remained relatively constant, regardless of the number of transferred embryos per patient. The implantation rate per embryo, in relation to its morphology, was clearly lower when irregular blastomeres and fragments were present in the perivitelline area. Other embryos, regardless of their morphology, had an identical development potential. Analysis of the results of this series demonstrates the difficulty of determining the development potential of all the embryos on the basis of morphological criteria.
- Published
- 1991
- Full Text
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32. Epididymal sperm aspiration in conjunction with in-vitro fertilization and embryo transfer in cases of obstructive azoospermia.
- Author
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Bladou F, Grillo JM, Rossi D, Noizet A, Gamerre M, Erny R, Luciani JM, and Serment G
- Subjects
- Adult, Embryo Transfer, Epididymis cytology, Epididymis surgery, Female, Fertilization in Vitro, Humans, Male, Middle Aged, Oligospermia physiopathology, Ovulation Induction, Suction, Cell Separation methods, Oligospermia therapy, Spermatozoa
- Abstract
Epididymal sperm aspiration is a new treatment for vasal agenesis. In previous reports, epididymal spermatozoa resulted in pregnancy by utilizing in-vitro fertilization (IVF) or gamete intra-Fallopian transfer (GIFT). We sought to investigate the efficacy of epididymal sperm aspiration in conjunction with IVF in patients with congenital absence of the vas deferens or with secondary extended obstruction of spermatic ducts. Fifty-eight attempts were performed in 23 patients (25-50 years). Eight patients (34.7%) had vasal agenesis and 14 (60.8%) presented with vasal secondary extended obstruction. The sperm count was adequate (greater than or equal 20 x 10(6)/ml) in 13.8% of sperm retrievals and sperm motility of 20% was obtained in 15.5% of sperm retrievals. Fourteen attempts at IVF were performed with epididymal sperm counts of 2-44 x 10(6)/ml and motilities of 0-45%. A mean of six mature oocytes (0-13) were inseminated in each case. Five embryo transfers were performed in five patients' wives (35.7%) and two couples had an early pregnancy loss (14.2%). Epididymal sperm aspiration is an advance in treating such patients, as an adequate number of mature spermatozoa can be obtained and used for IVF. However, spermatozoa directly aspirated from the proximal epididymis and with fertilizing capacity in vitro, gave a high rate of embryo degeneration (greater than 50%) after embryo transfer.
- Published
- 1991
- Full Text
- View/download PDF
33. [Incidence of the morphological aspect on the rate of implantation of embryos obtained by fertilization in vitro].
- Author
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Grillo JM, Mercier GM, Vasserot M, De Victor B, and Vitry G
- Subjects
- Female, Humans, Embryo Implantation, Embryo Transfer, Fertilization in Vitro
- Abstract
1350 embryos obtained by In Vitro Fertilization (IVF) have been examined with the reverse microscope before their transfer in utero. Some embryos are "morphologically normal" (34.7%); some others are atypical. It is difficult to determinate the implantation rate by embryo according to morphological aspect because, most of the time we transfer several embryos to only one patient and we do not know, in case of pregnancy, which one or which ones have developed. Nevertheless, we have tried to appreciate, for 500 embryos transfers after IVF, the incidence of morphological aspect according to the presence or absence of "morphologically normal" embryos. The implantation rate is significantly higher in the groups in which you find "morphological normal" embryos than in the group where they are missing. (19.7% and 19.2% vs 10.2%). It seems that the aspect of the embryo when it is transferred, has an incidence on its future development.
- Published
- 1991
34. Nucleolar changes in human embryo during the pre-implantation stage. Activation of ribosomal genes during the nucleologenesis.
- Author
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Grillo JM, Vasserot M, Gamerre M, Vitry G, and Stahl A
- Subjects
- Autoradiography, Blastocyst ultrastructure, Blastomeres ultrastructure, Cell Nucleolus ultrastructure, Fertilization in Vitro, Gene Expression Regulation, Humans, Microscopy, Electron, RNA biosynthesis, Tritium, Uridine metabolism, Blastocyst physiology, Blastomeres physiology, Cell Nucleolus physiology, DNA, Ribosomal genetics
- Abstract
We studied the structural and functional organization of human embryo nucleoli during the pre-implantation stage from spare embryos obtained by in vitro fertilization. In human embryo nucleoli, structural modifications occur during the first cleavages. They lead to the constitution of a reticulated nucleolus from an initial structure called primary nucleolus. They appear during the third cleavage and correspond to the nucleogenesis. Autoradiography shows no transcription in the primary nucleolus. Transcription of rDNA starts on the periphery of the initial structure after the 4-cell stage. It corresponds to the beginning of the embryonic gene expression. The entire nucleolus will be progressively concerned by this transcription during the reticulation. Silver staining at the ultrastructural level shows that Ag-NOR proteins are missing in the primary nucleolus. In the beginning of nucleogenesis, Ag-NOR proteins are first located on the nucleolar periphery. Their following distribution corresponds to the structures containing rDNA in the nucleolus. Nucleologenesis in the human embryo during the pre-implantation stage first requires an association of rDNA with a primary structure, then an activation of ribosomal genes.
- Published
- 1991
- Full Text
- View/download PDF
35. [The fertilization potential of sperm].
- Author
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Grillo JM
- Subjects
- Female, Humans, Male, Sperm Count methods, Sperm-Ovum Interactions, Fertilization, Spermatozoa physiology
- Abstract
In vitro fertilization (IVF) has made it possible for an increase in the knowledge of the potential of sperms to fertilize. IVF has confirmed that the classical parameters accepted in assessing spermograms does not constitute a sure way of predicting whether a subject is fertile or sterile. It is necessary to increase and redefine these data with new parameters. New techniques have been developed during the last few years to assess the functional quality of sperm and their efficiency so that in future they can be correlated with sperm fertility at least within the context of in vitro fertilization. The precise mechanisms that are responsible for infertility are still badly understood; so that it will be valuable to study at the cellular level interactions between spermatozoa and oocytes. This article describes some of the elements that could make it possible to assess sperm fertilizing capacity.
- Published
- 1991
36. [Ultrastructure of the rat neurohypophysis following electric stimulation of the supraoptic nuclei].
- Author
-
Grillo JM, Conte-Devolx B, and Vitry G
- Subjects
- Animals, Cytoplasmic Granules ultrastructure, Electric Stimulation, Electrodes, Implanted, Inclusion Bodies ultrastructure, Male, Pituitary Gland, Posterior physiology, Rats, Stereotaxic Techniques, Hypothalamus physiology, Neurosecretion, Pituitary Gland, Posterior ultrastructure, Supraoptic Nucleus physiology
- Abstract
By means of electrical stimulation of the supraoptic nuclei, in a stereotaxic apparatus, partial or complete degranulation of the neurohypophysis is obtained, with often a continued presence of large vesicles; the number of microvesicles is always increased.
- Published
- 1975
37. [Effect of hypoxia and of anoxia on the posterior pituitary gland of the rat].
- Author
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Roux C, Capuccio P, and Grillo JM
- Subjects
- Animals, Behavior, Animal, Cytoplasmic Granules, Male, Neurosecretion, Pituitary Gland, Posterior ultrastructure, Rats, Hypoxia complications, Pituitary Gland, Posterior physiology
- Abstract
The results of the effect of hypoxia or anoxia upon the rat neurohypophysis shows a decrease of the neurosecretory granules and the rise of the small vesicles.
- Published
- 1975
38. [Cyto-chemical study of the uterine cervix (author's transl)].
- Author
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Vitry G, Roux C, and Grillo JM
- Subjects
- Biopsy, Cervix Uteri cytology, Cervix Uteri pathology, Female, Histocytochemistry, Humans, Pregnancy, Cervix Uteri enzymology, Uterine Cervical Diseases enzymology
- Published
- 1976
39. Binding of 125I-toxin, from the spitting cobra Naja mossambica mossambica, at frog neuromuscular junctions.
- Author
-
Tessier M, Grillo JM, and Mambrini J
- Subjects
- Animals, Anura, Autoradiography, In Vitro Techniques, Iodine Radioisotopes, Receptors, Cholinergic metabolism, Cobra Neurotoxin Proteins metabolism, Elapid Venoms metabolism, Neuromuscular Junction metabolism
- Published
- 1978
- Full Text
- View/download PDF
40. [Effects of the injection of kainic acid into the medial hypothalamus on the secretion of corticolipotropic hormones in the rat].
- Author
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Grillo JM, Oliver C, Giraud P, Conte-Devolx B, Mercier G, and Vitry G
- Subjects
- Animals, Injections, Male, Rats, Rats, Inbred Strains, beta-Endorphin, Adrenocorticotropic Hormone metabolism, Endorphins metabolism, Hypothalamus drug effects, Hypothalamus, Middle drug effects, Kainic Acid pharmacology, Melanocyte-Stimulating Hormones metabolism, Pyrrolidines pharmacology
- Abstract
alpha-MSH secretion was significantly lowered in Rats after lesion of the mediobasal hypothalamus with microinjections of kainic acid. beta-Endorphin release after ether-stress was also reduced. In opposition, no significant change in basal and ether-stress ACTH secretion was observed.
- Published
- 1981
41. [Estimation of the quality of embryos obtained during fertilization in vitro as a function of their morphology].
- Author
-
Grillo JM, Mercier GM, Vasserot M, Luciani JM, and Vitry G
- Subjects
- Embryo Implantation, Embryo Transfer, Embryonic and Fetal Development, Female, Humans, Pregnancy, Retrospective Studies, Embryo, Mammalian anatomy & histology, Fertilization in Vitro methods
- Abstract
Embryo quality after IVF should allow provisions for their pregnancy potential. We have classified all the transferred embryos into 4 types according to morphological criteria and we have studied the implantation rate of the different types. The results show: That pregnancies have been obtained with the 4 types of embryos. That it has been impossible to demonstrate statistically significant differences between the different types of transferred embryos even if embryos morphologically normal seem to be more favourable than others to induce a pregnancy. That the average implantation rate for each transferred embryo is of 14.4%.
- Published
- 1987
42. [Effects of electric stimulation of the para-ventricular nucleus on corticotropin function in Long-Evans and Brattleboro rats].
- Author
-
Rey M, Conte-Devolx B, Grillo JM, Castanas E, Mercier G, Boudouresque F, Giraud P, Millet Y, and Oliver C
- Subjects
- Adrenocorticotropic Hormone blood, Animals, Corticotropin-Releasing Hormone physiology, Electric Stimulation, Male, Rats, Rats, Brattleboro, Rats, Inbred Strains, Species Specificity, Adrenocorticotropic Hormone metabolism, Corticosterone metabolism, Paraventricular Hypothalamic Nucleus physiology
- Abstract
Two peptides have a strong corticotropin releasing factor (CRF) activity: arginine vasopressine (AVP) and a 41-residue peptide (41-CRF). Both peptides are present in high concentration in the PVN of the Rat, a zone of the hypothalamus at which electrical stimulation elicits ACTH release. Since homozygous Brattleboro Rats (Di/Di) congenitally lack endogenous AVP, it was of interest to compare the ACTH and corticosterone release after electrical stimulation of the PVN in Long-Evans (L.E.) and Di/Di Rats. In both L.E. and Di/Di Rats, there is a significant increase in plasma ACTH and corticosterone after electrical stimulation of the PVN. However, corticosterone levels are significantly lower in Di/Di than in L.E. Rats whether the Rats have been stimulated or not. It is concluded that a physiological CRF activity is present in the PVN of Di/Di Rats independently of the CRF like activity of AVP.
- Published
- 1982
43. [Cytological diagnosis of ruptured membranes (author's transl)].
- Author
-
Grillo JM and Vitry G
- Subjects
- Cytodiagnosis methods, Female, Histocytochemistry, Humans, Pregnancy, Staining and Labeling, Fetal Membranes, Premature Rupture diagnosis, Vaginal Smears
- Abstract
The authors have attempted to perform a simple and reliable test to affirm the diagnosis of a ruptured membranes. Various methods of staining and histochemical techniques were used. They conclude that the classical method for sampling and the alcohol-ether fixative are the best methods. Only the presence of transparent colourless squames, orange stained by The Harris-Shorr technique, is typical of fetal elements in the vaginal smears, especially they are grouped in clusters.
- Published
- 1980
44. [Localization of ribosomal genes and nucleolar activity in lymphocytes of swine (Sus scrofa domestica) stimulated by phytohemagglutinins].
- Author
-
Toga-Piquet C, Henderson AS, Grillo JM, Vagner-Capodano AM, and Stahl A
- Subjects
- Animals, DNA, Ribosomal, Karyotyping, Lymphocytes immunology, Nucleic Acid Hybridization, Phytohemagglutinins, Ribosomes physiology, Swine, Cell Nucleolus physiology, DNA genetics, Lymphocyte Activation, Lymphocytes physiology
- Abstract
The Pig chromosomes that contain rDNA sites displayed a polymorphism in the distribution of the genes among the nucleolar organizers located on pairs Nos. 8 and 10. Two, or more often three, active sites were observed in the chromosomes of lymphocytes stimulated with phytohemagglutinin. Only 5% of the metaphases showed a 4th small active site. At the onset of stimulation most cells contained one-two nucleoli; four nucleoli were never observed. After prolonged stimulation, the number of nuclei containing three nucleoli increased. A 4th small nucleolus appeared in a few cells, presumably formed by activation of the smallest rDNA site.
- Published
- 1984
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