Shirly Lahav-Baratz

ABIN-1 negatively regulates NF-κB by inhibiting processing of the p105 precursor

p105 plays dual roles in NF-kappaB signaling: in its precursor form it inhibits NF-kappaB activation, but limited processing by the ubiquitin system generates the p50 active subunit of the transcription factor. Here we show that ABIN-1, an A20-binding protein that is also known to attenuate NF-kappaB activation, inhibits p105 processing. p105 and ABIN-1 physically interact with one another, but the binding is not necessary for inhibition of processing. Rather, it appears to stabilize ABIN-1 and to increase its level, which further augments its inhibitory effect. Deletion of the processing inhibitory domain (PID) of p105 abrogates the inhibition which also requires the ABIN homology domain (AHD)-2 of ABIN-1. Together, the effects of ABIN-1 on p105 processing and of p105 on stabilizing ABIN-1 act to potentiate the NF-kappaB inhibitory activity of ABIN-1.

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.

Analyzing Factors Affecting the Success Rate of Frozen–Thawed Embryos

AbstractPurpose: In recent years the infertile population applying for IVF treatments was changed and so the indications for performing intracellular sperm injection (ICSI). The aim of this study was to analyze predicting factors of our thawing cycles. Methods: From December 1998 to July 2001, 440 consecutive thawing cycles were performed. Patient characteristics were examined. The number of cryopreserved embryos, number of transferred embryos, the timing of cryopreservation (48 h vs.72 h), and embryo survival rate were analyzed as a possible predictor for pregnancies achievement. Results: Conventional IVF patients characteristic was significantly different from ICSI population and analysis has been performed for every population separately. In the IVF population the women age, the number of transferred embryos, and timing of cryopreservation were factors significantly influencing the pregnancy rate. Interestingly, in the ICSI population only the number of transferred embryos was found to be a predictive factor. Conclusion: ICSI and IVF cycles should be analyzed separately. Not all the factors influencing the success rate in the conventional IVF population are valid in the ICSI population.

Habits of cell phone usage and sperm quality – does it warrant attention?

Male infertility constitutes 30-40% of all infertility cases. Some studies have shown a continuous decline in semen quality since the beginning of the 20th century. One postulated contributing factor is radio frequency electromagnetic radiation emitted from cell phones. This study investigates an association between characteristics of cell phone usage and semen quality. Questionnaires accessing demographic data and characteristics of cell phone usage were completed by 106 men referred for semen analysis. Results were analysed according to WHO 2010 criteria. Talking for ≥1 h/day and during device charging were associated with higher rates of abnormal semen concentration (60.9% versus 35.7%, P < 0.04 and 66.7% versus 35.6%, P < 0.02, respectively). Among men who reported holding their phones ≤50 cm from the groin, a non-significantly higher rate of abnormal sperm concentration was found (47.1% versus 11.1%). Multivariate analysis revealed that talking while charging the device and smoking were risk factors for abnormal sperm concentration (OR = 4.13 [95% CI 1.28-13.3], P < 0.018 and OR = 3.04 [95% CI 1.14-8.13], P < 0.027, respectively). Our findings suggest that certain aspects of cell phone usage may bear adverse effects on sperm concentration. Investigation using large-scale studies is thus needed.

The impact of early testicular sperm extraction or cryopreservation on the outcome of intracytoplasmic sperm injection--a randomized controlled study.

AbstractPurpose: To compare the outcome of sperm extraction 24 h before ovum pickup and on the day of oocyte retrieval. Methods: A controlled study was performed to compare the outcome of 90 sperm extractions and in vitro sperm injection cycles performed in 54 patients. Results: Available fresh sperm for the sperm injection procedure and cryopreservation obtained on the day of ovum pickup were similar to sperm collected 1 day before (33.3% vs. 39.4%, respectively). Fertilization rate obtained with fresh sperm was also similar (48.9% vs. 54%), respectively. Clinical pregnancy rate was 38% vs. 22% per embryo transfer, respectively (P = 0.235). When comparing an additional 24 cycles with cryopreservation of sperm retrieved on the day of ovum pickup, as well as a day previously, no significance was noted in the parameters. Conclusions: Sperm retrieved 24 h before oocyte retrieval and used as fresh or frozen–thawed for sperm injection are as effective as those used on the day of ovum pickup.

A20 inhibits both the degradation and limited processing of the NF-κB p105 precursor: A novel additional layer to its regulator role

p105 plays dual roles in NF-κB signaling: it generates the active subunit p50 and in its precursor form inhibits NF-κB activation. p105 processing occurs under basal conditions and increases following signaling. IκB kinase β (IKKβ) mediates phosphorylation at the C-terminal domain of p105, leads to accelerated processing and degradation of the precursor. A20 is a dual function deubiquitinating and ubiquitin ligating enzyme, involving in turning-off the NF-κB signaling pathway. Here we show that A20 suppresses TNFα-induced proteolysis of p105. We demonstrate that A20 inhibits both the signal induced processing and the degradation of p105 upstream to IKK stimulation. In addition, A20 represses the constitutive processing of the precursor protein in IKK independent manner. Silencing of A20 in cells by siRNA, restores p105 processing and the generation of p50. Functional analysis of A20, shows that the ubiquitin ligase activity mediated by its zinc finger domain (ZF), is required for the basal processing inhibitory effect. Its N-terminal ovarian tumor (OTU) domain, however, is not obligatory. We show that A20 inhibits p50 generation in cells by reducing the ubiquitination of its precursor, p105. Co-immunoprecipitation experiments show that A20 is immunoprecipitated by p105 only in the presence of its recently identified ligase, KPC1. Our data propose an additional novel mechanism to explain the known NF-κB inhibitory effects of A20: by affecting p105 ubiquitination and subsequently its degradation and limited processing.

p27 and its ubiquitin ligase Skp2 expression in endometrium of IVF patients with repeated hormonal stimulation

This preliminary study examined a possible effect of long duration repeated hormonal stimulation on the endometrium using a molecular tool. The expression of the hormone stimulated, cell cycle regulators, p27 and its ligase S-phase kinase-interacting protein2 (Skp2), were assessed in 46 endometrial samples of patients who underwent repeated IVF cycles (3-21). Skp2 protein is usually undetectable in normal tissue and can be demonstrated only in rapidly dividing cells. Samples from non-stimulated, normal cycling women served as control group A. Samples of endometrial carcinoma served as control group B. In secretory endometrium, the expression of p27 was found to be lower and Skp2 higher in the study group compared with control group A. Moreover, in 25% of patients of the study group, Skp2 expression was significantly higher (P < 0.05) compared with control group A, reaching concentrations demonstrated in endometrial carcinoma. The findings of this study suggest that repeated hormone stimulation cycles may disrupt endometrial physiology, potentially towards abnormal proliferation. These changes in protein expression are described for the first time in IVF patients and should be further investigated.