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Biomedical subjects

Martha Dirnfeld

Publications and source records attributed to Martha Dirnfeld.

10 recordsLinked to original sources
PubMed2005

A multicenter study evaluating the flowcytometric-based kit for semen analysis.

OBJECTIVE: To evaluate the precision and accuracy of a flowcytometric-based semen analysis kit and its application in four fertility centers, as compared with routine microscopic evaluation. DESIGN: A prospective comparative study. SETTING: Four fertility centers, located in Israel and the United States. PATIENT(S): Patients referred to fertility clinic for sperm evaluation. INTERVENTION(S): The precision of semen analysis by both methods was evaluated by inter- and intra-technician studies. The accuracy of the sperm counts obtained was assessed by counting a diluted sample and comparing the obtained results with the expected results according to the dilution factor. MAIN OUTCOME MEASURE(S): Sperm count, morphology, motility, viability, white cell count, antisperm antibodies. RESULT(S): The flowcytometric-based kit for semen analysis is more precise and accurate than the manual routine method. Smaller coefficient of variance (CV%) with results were obtained by the flowcytometric-based kit for all semen parameters as compared with the routine manual methods. Sperm density values determined by the flowcytometric-based kit correlated better with the dilution factor. The agreement rates between the flowcytometric and manual methods are sperm count 90%, motility 82%, vitality 70%, round cell counts 78%, and sperm bound antibodies 100%. CONCLUSION(S): Semen analysis by flowcytometric-based kit is advantageous because of improved precision and accuracy compared with the routine method. It provides similar clinical information to that obtained by routine method for sperm count, motility, and antisperm antibodies, and more accurate results for round cell counts (labeled all subclasses) and vitality.

Flow Cytometry↗
PubMed2005

Safety of drugs used in assisted reproduction techniques.

Infertility may affect one in six couples; however, the development of the assisted reproduction technique (ART) created the opportunity for a large proportion of the infertile population to bear children. Pharmacological agents are routinely used in ART, and new ones are introduced regularly, with the aim of retrieving multiple oocytes to increase the prospect of pregnancy. The combinations of drugs that are used have specific adverse effects, but it is mostly the combined action of more than one agent that causes the greatest concern. The matter is complicated by the suspicion that some techniques in ART, for example intracytoplasmic sperm injection for severe male infertility problems (including azoospermia), may also contribute to the increase in adverse effects, especially congenital malformation. Gonadotropin releasing hormone (GnRH) agonists are widely used in controlled ovarian hyperstimulation. It may give rise to a short period of estradiol withdrawal symptoms and it may also lead to luteal phase deficiency. Similarly GnRHa antagonists, which have been recently introduced to control ovarian hyperstimulation, can lead to luteal phase deficiency and may cause some local injection site reactions. The more pure form of gonadotropin leads to less local injection site reactions and their main adverse effects are associated with the consequences of multiple ovulations. It has been proposed that gonadotropins may be a factor in the increasing risk of ovarian cancer and possibly breast cancer, but this has not been substantiated. Prion infection is another potential hazard, although no cases have been reported. Ovarian hyperstimulation syndrome is a well recognised complication of controlled ovarian hyperstimulation in ART. It is usually a result of recruitment of a large number of ovarian follicles. Efforts to minimise the incidence of this syndrome and its severity are now well developed. Congenital malformations are another possible adverse effect of fertility drugs, but it is more probable that the increase in congenital abnormality that is reported in ART is because of the population studied, i.e. patients already at high risk of congenital malformation, rather than the fertility drugs used or the technique employed. High order multiple pregnancy and its sequela is a well established complication of controlled ovarian hyperstimulation. This could be a result of multiple ovulations or more than one embryo replacement. Reducing the number of embryos transferred can reduce this more serious adverse effect for expectant mothers and for children conceived from ART.

Gonadotropins↗
PubMed2004

Oxidative stress indices in seminal plasma, as measured by the thermochemiluminescence assay, correlate with sperm parameters.

OBJECTIVE: To examine the effect of oxidation of proteins and lipids, as measured by a novel thermochemiluminescence (TCL) analyzer, and to evaluate the correlation between TCL indices in seminal plasma and sperm parameters. DESIGN: Experimental and prospective clinical studies. SETTING: An infertility unit. PATIENT(S): One hundred forty-eight men undergoing semen analysis. INTERVENTION(S): Bovine serum albumin (BSA) and linolenic acid were oxidized and tested by TCL, protein carbonyls, and conjugated dienes assays. All participants underwent semen analysis. Seminal plasma was tested by TCL and conjugated dienes. MAIN OUTCOME MEASURE(S): Thermochemiluminescence indices before and after oxidation of BSA and linolenic acid, compared with protein carbonyl and conjugated dienes indices. Correlation between semen parameters and TCL and conjugated dienes indices in seminal plasma. RESULT(S): Oxidation of BSA and linolenic acid was marked by characteristic changes in their TCL curve pattern and an increase in the levels of protein carbonyls and conjugated dienes. Among 125 sperm-containing semen samples, the TCL curve exhibited two patterns: a positive relative ratio curve (group A, 87 patients) and a negative relative ratio curve (group B, 38 patients). Sperm concentration was lower and total motile sperm and rapid motile sperm were fewer in group B. A significant correlation was found between TCL indices, conjugated dienes, and sperm quality in group B. CONCLUSION(S): Oxidation affects TCL curve pattern of proteins and lipids in a characteristic manner. Thermochemiluminescence indices in seminal plasma closely correlate with sperm characteristics among patients with sperm disturbances, and it might serve as a tool in the evaluation, treatment, and monitoring of subfertile men.

Animals↗
PubMed2004

Oxidative stress indices in follicular fluid as measured by the thermochemiluminescence assay correlate with outcome parameters in in vitro fertilization.

OBJECTIVE: To evaluate oxidative stress indices in follicular fluid (FF) by a novel thermochemiluminescence (TCL) assay and investigate the correlation between TCL and i.v.f. cycle parameters. DESIGN: Prospective, cross-sectional study. SETTING: I.v.f. Unit of an Obstetrics and Gynecology Department in a university-affiliated hospital. PATIENT(S): One hundred eighty-nine women undergoing consecutive i.v.f. treatment cycles during 2001. INTERVENTION(S): After oocyte retrieval, pooled FF was centrifuged and the supernatant was tested in the TCL assay. MAIN OUTCOME MEASURE(S): Maximal serum E2 levels, number of gonadotropin ampoules, retrieved oocytes, mature oocytes, fertilization and cleavage rates, number of available embryos and cryopreserved embryos, and clinical pregnancy rates and correlation with TCL indices. RESULT(S): The TCL curve slope of FF positively correlated with maximal serum E2 levels, number of mature oocytes, and number of cleaved embryos and was inversely correlated with the women's ages and the number of gonadotropin ampoules. Follicular fluid TCL amplitude at 50 seconds ranged from 294 to 711 cps, but all pregnancies (n = 50; 28.1%) occurred within the range of 347-569 cps. With 385 and 569 cps as cutoff levels for the occurrence of pregnancy, the negative predictive value beyond this range was 96% and the positive predictive value within this range was 32%. CONCLUSION(S): The TCL results may reflect the age-related increase in free radical activity and is associated with parameters of ovarian responsiveness and IVF outcome. A certain threshold of oxidative stress may be required for the occurrence of conception in i.v.f. TCL is a potential tool to evaluate, treat, and monitor antioxidant therapy in i.v.f. treatments.

Adult↗
PubMed2004

Early embryo-endometrial signaling modulates the regulation of matrix metalloproteinase-3.

OBJECTIVE: To examine matrix metalloproteinase-3 (MMP-3) expression in human stromal cell culture after P stimulation and the effect of conditioned medium from human embryo-epithelial cells coculture on its expression and activity. DESIGN: Metabolic and endocrine studies on human tissue. SETTING: In vitro fertilization (i.v.f.) unit and endocrine research unit. PATIENT(S): Infertile patients undergoing endometrial tissue sampling for dating at the luteal phase before i.v.f. INTERVENTION(S): Endometrial sampling and collection of human embryos culture media. MAIN OUTCOME MEASURE(S): Expression and activity of secreted MMP-3 by P-induced stromal cells, and in stromal cells exposed to conditioned medium from embryo-epithelial cell coculture. RESULT(S): Expression and activity of MMP-3 in human stromal cells decrease after P induction. Following incubation of these stromal-derived decidual cells with conditioned medium from embryo-epithelial cell coculture, MMP-3 expression and activity increased in a statistically significant manner. CONCLUSION(S): Progesterone inhibition of MMP-3 expression and its support of endometrial integrity were prevented by local expression of MMP-3 in response to embryonic signaling.

Adult↗
PubMed2003

Decreased expression of tissue inhibitor of matrix metalloproteinases in follicular fluid from women with polycystic ovaries compared with normally ovulating patients undergoing in vitro fertilization.

OBJECTIVE: To compare activity of matrix metalloproteinases (MMP) and expression of their tissue-specific inhibitor (TIMP) in the follicular fluid of normally ovulating women and women with the polycystic ovary syndrome (PCOS). DESIGN: Prospective study. SETTING: IVF unit and endocrine research unit. PATIENT(S): Fourteen patients undergoing IVF treatment (seven with normal ovulation and seven with PCOS). MAIN OUTCOME MEASURE(S): Activity of MMP-2 and MMP-9 and expression of MMP-1, TIMP-1, and TIMP-2 was measured in follicular fluid of the leading follicles by using gel zymography and immunoblot analysis. RESULT(S): The activity of MMP-2 and MMP-9 and expression of MMP-1 was similar in follicular fluid of normally ovulating patients and patients with PCOS. Significantly lower expression of TIMP-1 was found in follicular fluid of patients with PCOS women compared with normally ovulating patients. CONCLUSION(S): Because MMPs and TIMPs play a role in the physical and chemical structure of the follicular compartment, the decreased expression of TIMP in patients with PCOS may be part of a compensatory process to overcome the physical properties of the thick ovarian capsule.

Adult↗
PubMed2003

Morphometrical quantification of spermatogonial germ cells with the 57B anti-MAGE-A4 antibody in the evaluation of testicular biopsies for azoospermia.

The melanoma-associated antigen (MAGE) gene family of cancer-testis antigens is expressed in certain malignant neoplasms and the testis, but not in other healthy tissues. The aim of this study was to determine the usefulness of immunohistochemical staining with the 57B anti-MAGE-A4 mouse monoclonal antibody (MAb) in testicular biopsy specimens from patients with nonobstructive azoospermia and obstructive azoospermia (OA). Fifty-four cases of Sertoli cell only (SCO), 30 cases of spermatocytic arrest, 15 cases of hypospermatogenesis, and 10 testicular biopsy specimens with OA (normal spermatogenesis) were evaluated. Immunohistochemistry was performed using the 57B MAb, which primarily recognizes the MAGE-A4 antigen in paraffinized tissues. The cells were quantitated by a computerized image analysis system. Testicular biopsy specimens with normal spermatogenesis exhibited strong nuclear and cytoplasmic MAGE-A4 staining of spermatogonia and weak staining of spermatocytes, but not spermatids or Sertoli or Leydig cells. No staining was detected in SCO cases. In five cases of SCO with focal spermatogenesis, spermatogonial cells that were initially missed by hematoxylin and eosin staining were detected by MAGE-A4 immunohistochemistry. Immunostaining with the 57B MAb greatly enhanced identification of spermatogonia in cases of spermatocytic arrest and hypospermatogenesis. The number of MAGE-A4-positive spermatogonia was significantly decreased in hypospermatogenesis, as opposed to the OA group (12.1 +/- 4.3 and 30.3 +/- 10.0, respectively). The number of MAGE-A4-positive primary spermatocytes was significantly increased in early maturation arrest, as compared with the OA group (48.2 +/- 10.8 and 16.9 +/- 9.8, respectively). The 57B anti-MAGE-A4 MAb is a useful marker for the detection and quantitation of spermatogonial germ cells. It also facilitates automated image analysis and provides greater accuracy in the histopathologic evaluation of testicular biopsy specimens.

Adolescent↗
PubMed2003

Inverse relationship between Skp2 ubiquitin ligase and the cyclin dependent kinase inhibitor p27Kip1 in prostate cancer.

PURPOSE: Prostate carcinomas show a low level of the cell cycle inhibitor p27, which correlates with tumor aggressiveness. In tumors p27 is of the WT species and its deregulation is due to aberrant ubiquitin mediated degradation. The p27 is degraded following phosphorylation and subsequent recognition by SCFSkp2 ubiquitin ligase. We examined the relationship between p27 and its specific ligase Skp2 in normal and malignant prostate tissues. A possible correlation among the levels of these proteins, tumor grading and clinical state was also investigated. MATERIALS AND METHODS: Using immunohistochemistry immunofluorescence microscopy and Western blot analysis 51 samples from needle biopsies, transurethral resection and radical prostatectomy were analyzed for p27 and Skp2 expression. Correlation with tumor grading (Gleason) was performed. In 22 proven metastatic or organ confined cases correlation was also done with the Ki67 proliferative marker. RESULTS: Skp2 expression demonstrated a significant and direct correlation with malignancy (p <0.0001). Furthermore, a significant correlation was found between Skp2 level and tumor aggressiveness graded by Gleason score (p <0.0002) and prostate specific antigen. Patients with metastases had significantly higher Skp2 and Ki67 expression than those with organ confined disease (p <0.0001). In addition, Skp2 levels significantly correlated with Ki67 (r = 0.73, p <0.0001). An inverse correlation was found between p27 and Skp2 ligase. CONCLUSIONS: Skp2 expression in prostate biopsies may be used as an additional marker for tumor aggressiveness. The results also suggest a role for Skp2 in the pathogenesis of prostate malignancy.

Aged↗
PubMed2002

Immunosuppressive properties of follicular fluid and media conditioned by zygotes correlate with subsequent conception in in vitro fertilization.

PROBLEM: Our objective in this study was to correlate immunosuppressive properties of follicular fluid (FF) and media conditioned by zygotes and early embryos with the occurrence of conception in an in vitro fertilization (IVF) program. METHOD OF STUDY: Fifty-seven IVF patients were studied. Donor lymphocytes were incubated with mitogens and FF from mature oocytes or conditioned media from zygotes and early embryos. Proliferation was assessed by radioactive thymidine incorporation. Proliferation Index (PI) was the ratio between radioactive labeling of lymphocytes incubated in the presence and absence of a mitogen. RESULTS: The FFs and media conditioned by zygotes from conception cycles had higher immunosuppressive activity than those from non-conception cycles. CONCLUSIONS: Immunosuppressive activity present in FF and media conditioned by zygotes may be a major determinant of conception in IVF, and may serve as a marker for embryo quality.

Culture Media, Conditioned↗