Ibrahim M. Seraj

Human papillomavirus gene sequences in washed human sperm deoxyribonucleic acid

The present study demonstrated the presence of HPV gene sequences in Percoll-washed sperm cells using polymerase chain reaction primers targeting smaller gene regions. Up to 64% of the sperm specimens were shown to contain gene sequences indicative of the presence of HPV. Human papillomavirus type 16 was detected about twice as often as HPV type 18. The results suggest the possible role of sperm as a vector for HPV.

Stromal invasion in endometrial adenocarcinoma

The ability to predict myometrial invasion by endometrial adenocarcinoma is useful for preoperative treatment purposes. The presence of stromal invasion is one possible method of predicting myometrial invasion, but criteria for the presence of stromal invasion have been lacking. Recently, criteria for its detection were proposed. We evaluated the validity of the proposed criteria in two conditions: (1) atypical hyperplasia (119 cases) and (2) endometrial adenocarcinoma Stage I, grade 1 (83 cases). Adenocarcinoma without stromal invasion according to the criteria of Kurman and Norris was present in 43 cases and adenocarcinoma with stromal invasion was present in 40 cases. This series demonstrated the ability of prehysterectomy endometrial sampling to predict the probability of myometrial penetration. In addition, none of our patients without stromal invasion in the resected uterus had myometrial penetration and no metastases have occurred. In those showing stromal invasion in the excised uterus, the myometrium was invaded in 34 of 51 cases (67%). Since myometrial penetration was not present when there was no stromal invasion in the resected uterus, stromal invasion deserves further evaluation and should be considered in future classification of endometrial adenocarcinoma.

An analysis of endometrial biopsies performed for infertility

The authors evaluated 774 endometrial biopsies that were performed for infertility. Complications arose in 3.6%. Lag of more than 2 days was found in 19%; luteal phase defect (LPD) was diagnosed in 5.7%. Most of the incidence of LPD can be predicted from chance occurrence. There was no association between abnormal biopsies and basal body temperature patterns, or between pathology, pregnancy outcome, and treatment. Exceptions included women with multiple spontaneous abortions and patients treated with clomiphene citrate (CC). An endometrial biopsy was performed in a pregnancy cycle in 4.0%, with an abortion rate not significantly different from the total study group. The authors conclude that an endometrial biopsy is relatively safe; however, the diagnostic and therapeutic consequences are limited. Endometrial biopsies may be useful only if performed in cases of habitual abortion or ovulation induction with CC.

Evidence for ease of transmission of human papillomavirus DNA from sperm to cells of the uterus and embryo

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Microinvasive squamous carcinoma of the vulva: present status and reassessment.

A retrospective analysis of 24 patients with early invasive squamous carcinoma was performed. No nodal metastases were noted in these patients. Based on a review of the literature, no absolute definition of microinvasive carcinoma could be formulated, but a treatment outline has been formulated based on depth of invasion for Stage I lesions.

Detection of exogenous DNA in blastocysts after continuous exposure to DNA carrier sperm.

Recently, it was demonstrated that sperm cells could be used to carry foreign DNA or exogenous DNA fragments through an artificial reproductive tract and transfer the DNA to mouse blastocysts at the other end of the tract (1). Using radiolabeling methods, the DNA fragments have been reported to be localized at the postacrosomal and equatorial regions of the sperm head (2,3). There is some evidence of internalization of the DNA fragments into the compact sperm head (3). However, the majority of the foreign DNA pieces appears to be localized externally on the membrane surface and the exchange of DNA fragments between the sperm plasma membrane and the plasma membrane of hatched blastocyst cells proceeded with fluidity (1). This represents a potential problem in vivo for the inadvertent transfer of viral and bacterial DNA from the ejaculated sperm to the preimplantation blastocysL This is particularly of concern when the blastocyst is still in the reproductive tract or uterus of a patient in the early stages of pregnancy. In this study, the null hypothesis is that the exogenous DNA transferred from the sperm to the blastocyst cells does not become integrated with the host genome and is destroyed by intrinsic DNAse enzymatic activity. The objective was to determine the fate of the internalized exogenous DNA in mouse blastocysts after brief or continuous exposure to DNA carrier sperm in vitro. The cytoplasmic and nuclear components of the cells were sepaMATERIALS AND METHODS

Uptake of exogenous human papilloma virus L1 DNA by oocytes and detection by the polymerase chain reaction

ObjectiveThe purpose of this study was to determine if oocytes were capable of taking up exogenous DNA such as human papillomaviral (HPV) DNA and evaluate the zona pellucida as a barrier to the entry of foreign DNA into the oocyte.MethodsThe experiment consisted of four groups of hamster oocytes exposed to HPV DNA fragments: Group A, zona-free oocytes (n =5); Group B, oocytes with an intact zona pellucida (n =5); Group C, oocytes fixed in 4% buffered formalin solution for 20 min (n =5); and Group D, zona-free oocytes (n =4). Group C oocytes served as an internal control to ensure adequate washing of the oocytes after incubation.ResultsThe zona pellucida was not a barrier to foreign DNA molecules. The PCR did not detect L1-HPV and β-globin gene sequences in the untreated hamster oocyte. Uptake of the smaller DNA fragments such as that amplified from the β-globin region was independent of active oocyte cell processes.ConclusionOocytes cultured in vitro can passively take up exogenous DNA fragments. The results suggest a possible role of oocytes as vectors for foreign DNA.

Polymerase chain reaction enzyme-linked immunosorbent assay detection of mycoplasma consensus gene in sperm with low oocyte penetration capacity**Presented at the 52nd Annual Meeting of the American Society for Reproductive Medicine, Boston, Massachusetts, November 2 to 7, 1996.

OBJECTIVE To determine the prevalence of mycoplasmas in washed sperm and to compare the penetration of zona-free hamster oocytes by sperm with and without mycoplasmas. DESIGN Prospective comparative study. SETTING Clinical and academic research environment. PATIENT(S) Semen from 34 male patients. INTERVENTION(S) Specimens were divided, Percoll washed, and scanned for differences in kinematic parameters. Sperm DNA was extracted and assayed for mycoplasma DNA using the polymerase chain reaction-ELISA method targeting the consensus gene of 15 mycoplasma species. Remaining sperm were processed by centrifuge, Percoll, or TEST (TES and Tris) Yolk Buffer (TYB) and assessed for penetration capacity. MAIN OUTCOME MEASURE(S) Detection of mycoplasma DNA. RESULT(S) Mycoplasma DNA was detected in 29.4% of the Percoll-washed sperm. The penetration of oocytes by mycoplasma-positive sperm (59.5% +/- 17.3%; mean +/- SEM) was less than mycoplasma-negative sperm (86.8% +/- 5.4%) in the TYB-processed group. CONCLUSION(S) Mycoplasma DNA is demonstrated in almost a third of the Percoll-washed sperm. Because there were no other cell types except sperm, the results suggest that the mycoplasmas were either internalized or attached to the membranes. The reduced penetration by mycoplasma-positive sperm after 48-hour TYB suggest mycoplasmas required time to affect sperm function. Similarities between hypo-osmotic swelling and between kinematic parameters suggest that the mechanism does not involve differences in membrane integrity and in motility patterns.