A molecular survey of Israeli Duchenne and Becker muscular dystrophy patients.

Duchenne (DMD) and Becker (BMD) muscular dystrophy are allelic X-linked recessive diseases caused by a mutation in the dystrophin gene located on the short arm of chromosome X (Xp21). The dystrophin gene is the largest gene known in humans, extending over 2300 kb and containing more than 70 exons coding for a 420 KD protein comprising 3685 amino acids. The gene is highly unstable, with a high percentage of deletions and rearrangements. A third of dystrophin mutations are new mutations. The frequency of DMD is 1:3500 liveborn males, and that of BMD 1:10000. These dystrophies are severe, progressive, and lethal. BMD/DMD patients and 2/3 of female carriers have high levels of creatine phosphokinase (CK). During the past 5 years, 169 families with patients affected by progressive muscular dystrophy were examined and counselled. We were able to exclude the diagnosis of DMD/BMD in 49 families on the basis of clinical symptoms and signs, normal dystrophin on biopsy (11 families) and/or the absence of linkage to chromosome X by analysis of RFLP derived haplotypes. Molecular analysis was performed on 111 DMD/BMD families (five BMD and 106 DMD) with 81 available probands. This study resulted in the establishment in Israel of an integrated diagnostic protocol for DMD/BMD, employing genetic, biochemical and molecular techniques. Molecular analysis provided most of the families with new and essential information.

Ovulation induction with clomiphene and the rise in heterotopic pregnancies. A report of two cases.

Two women were treated for heterotopic pregnancies, the simultaneous occurrence of an intrauterine pregnancy and an ectopic pregnancy. The commonly accepted incidence is 1:30,000. The actual rate appears to be significantly higher; the two most prominent reasons are today's increased rate of ectopic pregnancies and the increased use of clomiphene in infertile women. A rigorous evaluation is required in all early pregnancies in which an ectopic is suspected to rule out the presence of a heterotopic pregnancy.

Chlamydial antibody, as determined with an enzyme-linked immunosorbent assay, in tubal factor infertility.

Serum chlamydial antibody (CA), as determined with an enzyme-linked immunosorbent assay (ELISA), was evaluated as a predictor for the presence or absence of tubal factor infertility. Two hundred fifty-eight infertile women had CA drawn at the initial visit of an infertility workup. Of them, 46.3% were CA positive (CA+). One hundred forty-five patients underwent laparoscopy (LPY). Tubal factor was diagnosed in 87.2% of CA+ patients and 13.6% of CA negative (CA-) ones (P less than .001), with a rising frequency by CA positivity. CA correctly predicted the presence or absence of tubal factor in 86.9% of patients. The frequency of abnormal hysterosalpingograms (HSG) was higher in CA+ patients. The predictive values for tubal factor with low, mid and high CA+ were 62.5%, 97.5% and 95.8%, respectively, and for no tubal factor with CA- was 72.3%. Combining HSG with CA- increased that value. Agreement between the LPY and HSG findings by the CA result showed a high correlation. A history of pelvic inflammatory disease (PID) or intrauterine device use was more common in CA+ patients, but only 25.3% of patients with tubal factor had a history of PID. The frequency of positive cervical chlamydial cultures was 0.8%. CA determined with ELISA appears to be an accurate screening test for tubal factor infertility and can be used to reliably select the procedure of choice for tubal evaluation.

Zona pellucida-mediated acrosomal exocytosis in mouse spermatozoa: characterization of an intermediate stage prior to the completion of the acrosome reaction.

The zona pellucida (ZP)-induced acrosome reaction in mouse sperm proceeds in two steps, identified by three sperm fluorescence patterns observed sequentially with the fluorescent probe chlortetracycline. Capacitated, acrosome-intact sperm displaying a B pattern proceed to an intermediate S pattern, and then progress from the S pattern to the fully acrosome-reacted AR pattern. Previously, it was not feasible to characterize the nature of the transient intermediate S pattern. Recently, it was demonstrated that sperm bind to the ZP of eggs treated with 12-O-tetradecanoyl phorbol-13-acetate (TPA) and undergo a B to S transition, but do not complete the acrosome reaction. These cells accumulate in the S pattern and fail to undergo the S to AR transition (Endo, Y., Schultz, R. M., and Kopf, G. S. 1987a. Dev. Biol. 119, 119-209). The present study utilized ZP from TPA-treated eggs to assess the state of S pattern sperm. The kinetics of the B to S transition of sperm incubated with either structurally intact or solubilized ZP from untreated or TPA-treated eggs are identical. Addition of either solubilized ZP from untreated eggs or A-23187 to S pattern sperm bound to intact or solubilized ZP from TPA-treated eggs induces the S to AR transition, while ZP from TPA-treated or fertilized eggs does not. Loss of the transmembrane pH gradient in the anterior portion of the sperm head, monitored by the fluorescent pH probe 9-N-dodecyl aminoacridine, follows the B to S transition in sperm incubated with ZP from unfertilized eggs, but no loss is observed when the B to S transition is induced using ZP from TPA-treated eggs. Subsequent addition of solubilized ZP from untreated eggs or A-23187 results in the loss of the transmembrane pH gradient of these S pattern sperm. Addition of nigericin to S pattern sperm bound to ZP from TPA-treated eggs discharges the transmembrane pH gradient and causes the S to AR transition. In contrast, nigericin added to B pattern sperm discharges the pH gradient but does not induce a B to S transition. Electron microscopic evaluation of S pattern-arrested sperm using ZP from TPA-treated eggs reveals intact plasma and outer acrosomal membranes. These results suggest that ZP from TPA-treated and fertilized eggs are modified such that the ZP ligands inducing the S to AR transition are lost or are inactivated.(ABSTRACT TRUNCATED AT 400 WORDS)

Transcervical fallopian tube recanalization: a safe and effective therapy for patients with proximal tubal obstruction.

Over a 13-month period, 14 patients with proximal tubal obstruction underwent transcervical fallopian tube recanalization under fluoroscopic guidance in an outpatient setting at the hospital of the University of Pennsylvania. Twenty-one of 24 attempted tubal dilations (87.5%) were successful, as demonstrated by tubal opacification and contrast spillage into the peritoneal cavity at the conclusion of the procedure. Four intrauterine pregnancies, and no ectopic pregnancies, have followed the recanalization. One pregnancy ended in an early miscarriage, one patient delivered a healthy term female, and two pregnancies are ongoing at greater than twenty weeks' gestation. Two procedure-related complications occurred: in one patient, the isthmic segment of a fallopian tube was perforated, but healed without incident, and another patient experienced a low-grade fever, which resolved with p.o. antibiotics. We therefore conclude that fallopian tube recanalization is a well-tolerated, safe, and effective procedure for the treatment of proximal tubal occlusion.

Immunocytochemical and biochemical characterization of guanine nucleotide-binding regulatory proteins in mammalian spermatozoa.

Polyclonal antisera directed against conserved and subtype-specific peptide sequences of the alpha-subunits of guanine nucleotide-binding regulatory proteins (G proteins) were used to characterize the nature of mammalian sperm G proteins and to determine whether their localization was consistent with their proposed roles in mediating ZP3-induced acrosomal exocytosis. Mouse and guinea pig sperm exhibit positive immunofluorescence in the acrosomal region using an antiserum directed against a peptide region common to all alpha-subunits of G proteins (G alpha). The immunofluorescence disappears after sperm have undergone the acrosome reaction, suggesting that the immunoreactive material is associated with the plasma membrane/outer acrosomal membrane region overlying the acrosome. The presence of G proteins in this region is confirmed by the presence of a Mr 41,000 substrate for pertussis toxin (PT)-catalyzed [32P]ADP-ribosylation in purified plasma membrane/outer acrosomal membrane hybrid vesicles obtained from acrosome-reacted guinea pig sperm. Immunoprecipitation and polyacrylamide gel electrophoresis of PT-catalyzed [32P]ADP-ribosylated protein(s) using anti-peptide antisera generated against sequences unique to Gi alpha 1, Gi alpha 2, and Gi alpha 3 confirm the existence of all three Gi subtypes in mouse sperm extracts. Indirect immunofluorescence using an antiserum directed against a peptide region present in Gz alpha, a PT-insensitive G protein, demonstrates positive immunoreactivity in the postacrosomal/lateral face region of the mouse sperm head. This immunoreactivity is retained during acrosomal exocytosis in response to solubilized ZP and then disappears subsequent to this exocytotic event. These data demonstrate that Gi protein alpha-subunits are present in the acrosomal region of mammalian sperm, consistent with their postulated role in regulating ZP3-mediated acrosomal exocytosis, and that PT-insensitive Gz alpha is found in a region of the sperm head distinct from that of the Gi alpha subunits.