Mary Lake Polan

Repetitive DNA sequences in Drosophila

The satellite DNAs of Drosophila melanogaster and D. virilis have been examined by isopycnic centrifugation, thermal denaturation, and in situ molecular hybridization. The satellites melt over a narrow temperature range, reassociate rapidly after denaturation, and separate into strands of differing buoyant density in alkaline CsCl. In D. virilis and D. melanogaster the satellites constitute respectively 41% and 8% of the DNA isolated from diploid tissue. The satellites make up only a minute fraction of the DNA isolated from polytene tissue. Complementary RNA synthesized in vitro from the largest satellite of D. virilis hybridized to the centromeric heterochromatin of mitotic chromosomes, although binding to the Y chromosome was low. The same cRNA hybridized primarily to the α-heterochromatin in the chromocenter of salivary gland nuclei. The level of hybridization in diploid and polytene nuclei was similar, despite the great difference in total DNA content. The centrifugation and hybridization data imply that the α-heterochromatin either does not replicate or replicates only slightly during polytenization. Similar but less extensive data are presented for D. melanogaster. — In D. melanogaster cRNA synthesized from total DNA hybridized to the entire chromocenter (α- and β-heterochromatin) and less intensely to many bands on the chromosome arms. The X chromosome was more heavily labeled than the autosomes. In D. virilis the X chromosome showed a similar preferential binding of cRNA copied from main peak sequences.—It is concluded that the majority of repetitive sequences in D. virilis and D. melanogaster are located in the α- and β-heterochromatin. Repetitive sequences constitute only a small percentage of the euchromatin, but they are widely distributed in the chromosomes. During polytenization the α-heterochromatin probably does not replicate, but some or all of the repetitive sequences in the β-heterochromatin and the euchromatin do replicate.

The use of high-dose human menopausal gonadotropin in an in vitro fertilization program

Sixty-three normal ovulatory women suffering from obstructive tubal disease not corrected by previous surgery were enrolled in an in vitro fertilization (IVF) program. To achieve a large number of mature follicles, a relatively high dose of human menopausal gonadotropin (hMG) was administered (19 +/- 4 ampules/cycle). Monitoring consisted of daily follicular ultrasonography and serum estradiol measurements. Human chorionic gonadotropin (10,000 IU) was administered when more than two large follicles (1.6 to 1.8 cm in diameter) were visualized. Fifty-five laparoscopies for oocyte retrieval were performed. A mean of 4.3 follicles per woman were aspirated, and 3.2 oocytes per woman were recovered. The oocytes were preincubated for 8 or 24 hours according to the morphologic degree of mucification and dispersal of the oocyte-corona-cumulus complex. Seventy-seven percent of the oocytes were fertilized and were transferred into the uterus 38 to 40 hours after insemination. Fifty-two women received one to eight embryos (mean, 3.5 +/- 1.9), and 9 (17%) conceived. This regimen of high-dose hMG precludes the need for serum or urine luteinizing hormone monitoring, because the occurrence of spontaneous ovulation is low. It is valuable in increasing the number of fertilizable oocytes, the percentage of women undergoing embryo transfer, and compensates with multiple oocyte transfer for the high embryonic loss involved in IVF.

The relationship between follicular fluid steroid concentration and successful fertilization of human oocytes in vitro

Follicular fluids (FF) and their matched oocytes were obtained from 64 follicles of 28 women who failed to conceive after in vitro fertilization ( IVF ) and 33 follicles of 8 women who successfully conceived after the procedure. Ovulation was induced with human menopausal gonadotropin, and follicular aspiration was performed 36 hours after human chorionic gonadotropin administration. The concentration of 17 beta-estradiol, progesterone, testosterone, dihydrotestosterone, and androsterone was correlated with the morphology of the oocyte-corona-cumulus complex ( OCCC ), oocyte fertilization, the rate of cleavage, and the incidence of pregnancy after embryo transfer. In both groups of women, FF progesterone was lowest in follicles containing immature OCCCs . However, follicles aspirated from women who conceived after IVF which contained intermediate and mature OCCCs had significantly higher FF estradiol levels than similar follicles from women who failed to conceive after the procedure. Fertilized oocytes and 4- to 6-cell stage embryos which were obtained from follicles of pregnant women contained significantly higher FF estradiol levels than fertilized oocytes and similar embryos from nonpregnant women. It appears that higher FF estradiol levels correlate well with successful fertilization and an enhanced cleavage rate of oocytes associated with pregnancy following IVF .

Resectoscopic management of müllerian fusion defects

Of the 103 patients taken to the operating room for hysteroscopic resection of a septum, 72 underwent successful resection of the septum. The initial hysterogram can identify those patients who may benefit from the procedure but cannot discriminate between bicornuate and septate uteri. Knowledge of the size and contour of the uterus is essential to the final decision to proceed transcervically, opposed to transabdominally. In the transcervical repair group, at the time this article was written, there were 58 term deliveries and 5 ongoing pregnancies greater than 30 weeks (1 had a spontaneous abortion after surgery and subsequently conceived a term pregnancy), for a successful pregnancy rate of 86%. This correlated well with the pregnancy rates observed after metroplasty. Therefore, hysteroscopic resection constitutes a valuable alternative to the classical transabdominal approach.

Satellite DNA sequences of Drosophila melanogaster

Abstract Three of the four major satellite DNAs of Drosophila melanogaster were isolated by centrifugation in Ag + Cs 2 SO 4 gradients. The satellites were characterized by buoyant density analysis in neutral and alkaline CsCl and by thermal melt analysis, and were judged to be pure by these criteria. Complementary RNA was synthesized using Escherichia coli RNA polymerase, [α-32P]NTPs, and the separated satellite DNA strands as template. The complementary RNA was analyzed by RNA sequencing techniques. The sequences determined for the satellites are simple in composition and are related to one another. Satellite I is composed of A-A-T and A-T in the molar ratio of 2A-A-T:3A-T. Satellite II consists of the ten-nucleotide repeating sequence: A-A-T-A-A-C-A-T-A-G. Satellite IV is composed of A-A-G, A-G and G in the molar ratios of 3A-A-G: 4A-G: IG. One sequence present in long regions of satellite IV is A-A-G A-G. The sequences of the three satellites may be represented by the general expression (AAN)m(AN)n. The similarity of the sequences provides evidence for theories concerning the origin of families of satellite sequences.

The periovulatory and luteal phase of conception cycles following in vitro fertilization and embryo transfer

The pattern of periovulatory and luteal phase levels of serum estradiol (E2) and progesterone (P) were compared between 8 conception and 28 nonconception cycles of patients undergoing in vitro fertilization (IVF). Ten additional women served as control subjects and did not undergo follicular aspiration. Follicle growth was induced with an individualized Pergonal (human menopausal gonadotropin) regimen, and laparoscopy was performed 36 hours after human chorionic gonadotropin administration. The length of the luteal phase did not differ significantly among the three groups and was between 14 and 15 days in duration. When IVF conception cycles were compared with nonconception cycles, although no difference in the number of large follicles was observed (4.25 +/- 0.45 versus 3.6 +/- 0.25), the patterns of E2 and P differed significantly. Daily serum E2 levels tended to be higher in the periovulatory phase in conception cycles when compared with nonconception cycles, and were significantly (P less than 0.05) higher in the early, mid, and late luteal phases. Serum P levels were significantly higher (P less than 0.05) in conception cycles from the midluteal phase onward. A decline in both serum E2 and P in the midluteal phase in conception cycles suggested some degree of corpus luteum deficiency. It is suggested that high E2 levels in the periovulatory phase may be an indicator of better follicular development under human menopausal gonadotropin stimulation and that the deficiency observed in the late luteal phase is overcome with the establishment of pregnancy.

Human menopausal gonadotropins: A justifiable therapy in ovulatory women with long-standing idiopathic infertility

Long-standing infertility of unknown etiology is currently an indication for in vitro fertilization. Because of the prolonged waiting time, expense, and operative risk, we have treated a group of women with empiric human menopausal gonadotropins/human chorionic gonadotropin before in vitro fertilization. A study group of 97 couples with well-documented, long-standing idiopathic infertility received up to four cycles of empiric human menopausal gonadotropin therapy before in vitro fertilization. This group was compared with a control group of 48 couples who did not receive human menopausal gonadotropin before in vitro fertilization. The 12 (12.4%) conceptions and eight (8.2%) term births resulting from empiric human menopausal gonadotropin therapy in the study group was significantly higher than the number of spontaneous conceptions and births (1%) in the study group (p less than 0.003) and was also greater than the spontaneous conception and birth rate (4%) in the control group (p less than 0.07). The conception rate with empiric human menopausal gonadotropin (12.4%) compared favorably with that after in vitro fertilization (7.9%) and 8 of 12 (66.7%) of empiric human menopausal gonadotropin conceptions resulted in term births, compared with two of seven (28.6%) of the in vitro fertilization conceptions. Preovulatory serum estradiol levels were similar for all study patients undergoing empiric human menopausal gonadotropin therapy, control patients undergoing in vitro fertilization, and patients receiving empiric human menopausal gonadotropin during conception cycles. It therefore appears appropriate to offer a 4-month trial of empiric human menopausal gonadotropin to that specific group of couples with long-standing idiopathic infertility.

Ovarian steroids modulate human monocyte tumor necrosis factor alpha messenger ribonucleic acid levels in cultured human peripheral monocytes

OBJECTIVE To determine whether tumor necrosis factor alpha (TNF-alpha) messenger ribonucleic acid (mRNA) levels in human peripheral monocytes are regulated by ovarian steroids. DESIGN Human granulosa-luteal cells and cultured, activated human peripheral monocytes were subjected to Northern blot analysis for TNF-alpha mRNA. SETTING Academic research laboratory. PATIENTS Two human female volunteers of reproductive age and in the luteal phase of the menstrual cycle. RESULTS Human granulosa-luteal cells produce TNF-alpha mRNA. Physiological levels of progesterone (P) and estradiol (E2) modulate TNF-alpha mRNA from peripheral blood monocytes with an apparent inverse relationship between steroid concentration and TNF-alpha message. CONCLUSIONS Progesterone and E2 at physiological concentrations regulate TNF-alpha mRNA production. The P antagonist mifepristone (RU486) and the E2 agonist/antagonist tamoxifen modulate total TNF-alpha mRNA levels, suggesting involvement of specific receptors.

The effect of interleukin-1 on adhesion formation. in the rat***

The potential role of interleukin-1 in postoperative adhesion formation was examined. Cecal abrasion gave a consistently higher adhesion score when compared with sham laparotomy, on the basis of adhesion number, density, and vascularity, and so was chosen for use in further studies. The extent of serosal bleeding during cecal abrasion did not affect adhesion scores. Intraperitoneal injection of 10 micrograms murine recombinant interleukin-1 alpha in cecally abraded animals on the day of surgery and on the following 4 days resulted in a significant increase in adhesion scores when compared with those of cecally abraded animals injected with vehicle alone. Adhesions enhanced with murine recombinant interleukin-1 alpha, which were thicker and more vascular, were equivalently enhanced at doses from 10 to 10,000 ng, implying maximal response over that range. Rats not operated on and receiving recombinant interleukin-1 alpha 2 weeks after injury had increased adhesion formation. These results demonstrate that interleukin-1 alpha may be an important short-term mediator of postsurgical adhesion formation.

The use of pure follicle-stimulating hormone for ovulation induction in normal ovulatory women in an in vitro fertilization program

Many ovulation induction protocols for follicular development have been reported. The present study examines pure follicle-stimulating hormone (pFSH) and human menopausal gonadotropin for ovulation induction in an in vitro fertilization and embryo transfer program. The study compares the number of ampules, the level of estradiol on the day of human chorionic gonadotropin administration and at laparoscopy, the number of oocytes retrieved, fertilization, cleavage, and pregnancy rates. The peak levels of estradiol on the day of human chorionic gonadotropin administration and the day of laparoscopy were similar, although fewer ampules of pFSH were required to reach similar criteria for oocyte maturation prior to retrieval. The fertilization rates were similar, but the cleavage and pregnancy rates favored the use of pFSH. The use of pFSH may be more physiologic in orchestrating follicular steroidogenesis in normal ovulatory women in an in vitro fertilization and embryo transplant program that subsequently could produce healthier oocytes and an improvement in the pregnancy rate.