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Biomedical subjects

A F Haney

Publications and source records attributed to A F Haney.

At least 19 recordsLinked to original sources

Inducible nitric oxide synthase expression by peritoneal macrophages in endometriosis-associated infertility.

OBJECTIVE: Determine whether peritoneal macrophages from women with endometriosis-associated infertility express more inducible nitric oxide synthase (NOS2) and produce more NO than fertile controls. DESIGN: Unblinded clinical study. PATIENT(S): Nine infertile women with endometriosis and nine normal fertile women undergoing laparoscopy. INTERVENTION(S): Peritoneal fluid and macrophages were collected. Cells were also cultured with the NOS2 inducers interferon-alpha (IFN-alpha) or IFN-gamma plus lipopolysaccharide (LPS). MAIN OUTCOME MEASURE(S): Peritoneal fluid NO levels, peritoneal macrophage NOS activity, and peritoneal macrophage NOS2 protein expression. RESULT(S): NOS enzyme activity was higher in peritoneal macrophages from endometriosis patients. Immunoblots demonstrated NOS2 protein only in peritoneal macrophages from women with endometriosis. Peritoneal fluid NO concentration was similar in the two groups, but total peritoneal fluid NO content was higher in endometriosis patients. After 3 days' culture, peritoneal macrophages from women with endometriosis produced more NO in response to IFN-alpha or IFN-gamma plus LPS than controls. CONCLUSION(S): Peritoneal macrophages from women with endometriosis-associated infertility express higher levels of NOS2, have higher NOS enzyme activity, and produce more NO in response to immune stimulation in vitro. As high levels of NO adversely affect sperm, embryos, implantation, and oviductal function, reducing peritoneal fluid NO production or blocking NO effects may improve fertility in women with endometriosis.

Animals↗

Malignancy in endometriosis: frequency and comparison of ovarian and extraovarian types.

One thousand consecutive cases of surgically proven endometriosis were reviewed to evaluate the frequency and types of pelvic cancers that were associated with ovarian and extraovarian endometriosis. The frequency and types of histologic abnormalities present in the eutopic endometrium when cancers were noted in endometriosis were also evaluated. In the large subset of cases for which the authors were the primary pathologists and all foci of endometriosis were recorded, the frequency of malignancy was 10.8%. In contrast, the frequency was only 3.2% in cases diagnosed by others previously in our institution. Cancers were more commonly found in ovaries when endometriosis was present in that ovary (5%) compared to when endometriosis was present at other sites (1%). Clear cell and endometrioid carcinomas were the malignancies most commonly seen in ovaries containing endometriosis, while clear cell adenocarcinoma and adenosarcoma were most commonly seen in conjunction with extraovarian endometriosis. The association of endometriosis with endometrioid and clear cell carcinoma was much stronger than that of serous and mucinous tumors (p < .01). Concurrent endometrial pathology was commonly seen in cases of malignant transformation of endometriosis (32% of cases).

Adenocarcinoma, Clear Cell↗

Identification of macrophages at the site of peritoneal injury: evidence supporting a direct role for peritoneal macrophages in healing injured peritoneum.

OBJECTIVE: To determine if peritoneal macrophages are present at the site of a surgical injury to the peritoneum during wound healing. DESIGN: Controlled research study. SETTING: Academic research laboratory. EXPERIMENTAL MODEL: A murine model of peritoneal wound healing. INTERVENTION(S): Intraperitoneal injection of polystyrene beads 1 hour after a surgical peritoneal injury to identify peritoneal macrophages. MAIN OUTCOME MEASURE(S): Presence of peritoneal macrophages at the site of the healing wound as determined by intracellular polystyrene beads on transmission electron microscopy 1, 3, 7, and 14 days after injury. RESULT(S): Peritoneal macrophages were easily distinguished from other cell types by the phagocytosis of polystyrene beads. One day after injury, peritoneal macrophages were adherent to the wound surface. By 3 days, mesothelial cells began covering the peritoneal macrophages at the wound surface and peritoneal macrophages were identified deep within the wound. Seven days after injury, the mesothelial layer was completely reconstituted, but peritoneal macrophages persisted within the healing would below the surface mesothelium. CONCLUSION(S): These data indicate that peritoneal macrophages are present at the peritoneal injury site throughout the healing interval and are consistent with these macrophages having a critical role in peritoneal wound healing.

Animals↗

Clinical decision making regarding leiomyomata: what we need in the next millenium.

Leiomyomata represent the most common gynecologic tumors and are responsible for over 200,000 hysterectomies per year. They are almost invariably benign and represent clonal expansion of individual myometrial cells. They can cause a variety of symptoms including menometrorrhagia, dysmenorrhea, pelvic pain, reproductive failure, and compression of adjacent pelvic viscera, or be totally asymptomatic. Leiomyomata are more common in African-American women and have a non-Mendelian inheritance pattern with up to a 50% recurrence rate after surgical removal. The therapeutic choices depend on the goals of therapy, with hysterectomy most often used for definitive treatment, and myomectomy when preservation of childbearing is desired. Intracavitary and submucous leiomyomata can be removed by hysteroscopic resection. Laparoscopic myomectomy is now technically possible but apparently with an increased risk of uterine rupture during pregnancy. Although gonadotropin-releasing hormone-agonist-induced hypogonadism can reduce the volume of leiomyomata, the severe side effects and prompt recurrences make them useful only for short-term goals such as reversing anemia or shrinking an intracavitary tumor prior to hysteroscopic resection. Nonextirpative approaches such as myolysis and uterine artery embolization are being evaluated, and may provide more options if they prove to be safe and efficacious in long-term follow-up. Ultimately, if the genetic basis for fibroid development and/or the molecular mechanism(s) of myometrial proliferation are understood, additional nonsurgical therapeutic interventions may be forthcoming. Current clinical needs are to a) determine an effective prevention strategy in genetically predisposed individuals; b) slow the growth of leiomyomata; c) identify the mechanisms of infertility; d) improve early detection; e) develop better surgical techniques; f) reduce recurrences after myomectomy; g) develop nonextirpative options; and h) evaluate their long-term results.

Decision Support Techniques↗

The temporal efficacy of early second-look lysis of adhesions in reducing postoperative adhesions in a murine model.

OBJECTIVE: Our aim was to determine whether early second-look lysis of adhesions reduces postoperative adhesions. STUDY DESIGN: With the murine uterine horn model, early second-look lysis of adhesions was performed 5, 7, 14, and 21 days after an electrocautery injury. Sites with adhesions (between 36 and 46/time point) underwent lysis of adhesions. Fourteen days later, a reviewer blinded to the treatment assessed adhesion formation, including adhesions not present at early second-look lysis of adhesions (eg, de novo adhesions). RESULTS: The rate of adhesion formation was 49% of control sites, unchanged when the early second-look lysis of adhesions was performed at 5 (44.4%) and 7 (39.5%) days, reduced at 14 days (28.6%), and increased at 21 days (74%). The pattern of de novo adhesions was similar, 17.6% when the early second-look lysis of adhesions was performed at 5 days, 10% at 7 days, 0% at 14 days, and 28.6% at 21 days. The only histologic difference between the groups was neovascularity at day 21. CONCLUSIONS: Early second-look lysis of adhesions was effective in reducing postoperative adhesions only when performed at 14 days in this model, suggesting that the specific cellular events occurring at the time of the early second-look lysis of adhesions are critical to efficacy.

Animals↗

A barrier composed of chemically cross-linked hyaluronic acid (Incert) reduces postoperative adhesion formation.

OBJECTIVE: To test whether a barrier of chemically cross-linked pure hyaluronic acid reduces postoperative adhesion formation. DESIGN: The material was evaluated in the murine uterine horn model using excision and electrocautery injuries and in animals who had amounts of material inserted into the abdomen to evaluate toxicity. SETTING: Academic medical center. SUBJECT(S): Mice. INTERVENTION(S): Insertion of the barrier between uterine horns and into the peritoneal cavity. MAIN OUTCOME MEASURE(S): Adhesion formation at 14 days; the histology of the peritoneum, liver, and spleen at 42 days; and the number, differential count, morphology, and flow cytometry of peritoneal leukocytes 3 days postoperatively. RESULT(S): Fewer adhesions were present when excision injuries were separated by the barrier (12 of 28 sites [43%] versus 23 of 26 control sites [88%]), whereas the number of adhesions was unchanged after electrocautery injuries (14 of 26 sites [54%] versus 17 of 26 control sites [65%]). The uterine horn sites covered by the barrier were histologically indistinguishable from controls. No adverse impact on the peritoneum and peritoneal fluid leukocyte population was observed with barrier insertion. CONCLUSION(S): The use of a barrier composed of a chemically cross-linked hyaluronic acid derivative (Incert, Anika Therapeutics, Inc., Woburn, MA) reduced postoperative adhesion formation in this model without any adverse impact on the peritoneum and peritoneal leukocyte population. This barrier material shows promise in preventing postoperative adhesions and deserves clinical evaluation.

Anesthesia, Intravenous↗

Removal of surgical barriers of expanded polytetrafluorethylene at second-look laparoscopy was not associated with adhesion formation.

OBJECTIVE: To evaluate whether the surgical trauma required for the laparoscopic removal of a polytetrafluoroethylene (PTFE) surgical barrier would cause postoperative adhesions. DESIGN: Two case reports. SETTING: Tertiary academic medical center. PATIENT(S): Two women who had undergone myomectomy with placement of a PTFE surgical barrier and who were free of adhesions with the barrier in place. INTERVENTION(S): Removal of the PTFE barrier by laparoscopy 11 days after myomectomy. MAIN OUTCOME MEASURE(S): Adhesions at the site of removal of the PTFE barrier at the time of incidental laparoscopy several years later. RESULT(S): Adhesions were not present at the site of PTFE barrier removal. CONCLUSION(S): The surgical trauma required to remove PTFE barriers at early second-look laparoscopy was not associated with postoperative adhesions.

Adult↗

Evaluation of the postcoital test in cycles involving exogenous gonadotropins.

OBJECTIVE: To evaluate the hypothesis that a postcoital test, optimally performed in the periovulatory period of cycles in which gonadotropin-induced superovulation was used, correlates with cycle fecundity. METHODS: Of 1135 total consecutive cycles, 367 first cycles were analyzed from the reproductive endocrinology and infertility service of a university medical center. This referral population had a mean age of 34.6 years for the female partner, a nulliparity rate of 81%, and a mean length of infertility of 4.8 years. Postcoital tests were performed 36-40 hours after hCG administration in gonadotropin-stimulated cycles. Clinical pregnancy was defined as fetal cardiac activity as seen on transvaginal ultrasound examination. RESULTS: Couples with no sperm observed per high-power field in the cervical mucus achieved a 16% fecundity rate (21 pregnancies in 129 cycles), one to ten sperm a 18% fecundity rate (28 pregnancies in 154 cycles), and more than ten sperm a 15% fecundity rate (13 pregnancies in 84 cycles). There was no significant difference between groups (n = 367, P = .85); the power to detect a statistically significant difference was .82. As validation of optimal cervical mucus, fecundity rates were compared with these postcoital test values across the entire range of peak periovulatory serum estrogen levels, and no correlation was seen (P = .61, .86, and .96 for estrogen levels of 201-500, 501-1500, and 1501-3433 pg/mL, respectively). CONCLUSION: With precise periovulatory timing and supraphysiologic estrogen levels optimizing qualitative cervical mucus characteristics in gonadotropin-induced cycles, the number of sperm observed per high-power field does not correlate with cycle fecundity.

Adult↗

Galactorrhea.

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Adult↗

The peritoneal response to expanded polytetrafluoroethylene and oxidized regenerated cellulose surgical adhesion barriers.

OBJECTIVE: To determine the peritoneal response to the surgical adhesion barriers expanded-polytetrafluoroethylene (ePTFE) and oxidized regenerated cellulose (ORC). STUDY DESIGN: The barriers were retrieved from the peritoneal cavities of women and mice 2 hours to 14 days after insertion and subjected to histology and electronmicroscopy. RESULTS: Macrophages and mesothelial cells rapidly appeared on the surface of both materials. ePTFE was covered by 3 days, with the macrophages gradually being replaced by mesothelium and disappearing thereafter. By 7 days, a delicate membrane with surface mesothelial cells completely enveloped the ePTFE, creating a "pseudoperitoneum". The membrane was difficult to recover as it was fragile and not adherent to the ePTFE. ORC was rapidly infiltrated and degraded by leukocytes and disappeared by 5 days in mice and from all but 1 of 20 women by 11 days. CONCLUSIONS: ePTFE is rapidly encapsulated by a non-adherent membrane resembling peritoneum while ORC is rapidly infiltrated and degraded by peritoneal fluid leukocytes.

Animals↗

Increased levels of laminin in ascitic fluid of patients with ovarian cancer.

Laminin is a component of the extracellular matrix and is associated with tumor cell metastasis. Present studies show that the ovarian cancer cell lines produce significant amounts of laminin (54-140 ng/ml) in culture. Since ovarian cancer is associated with ascites production, laminin levels were then determined in ascites and serum. The results indicate that the ascites from patients with serous adenocarcinoma of the ovary had higher levels of laminin than the normal peritoneal fluid (P < 0.0001). However, the serum levels of laminin did not differ significantly between the control population and ovarian cancer patients.

Ascitic Fluid↗

Nitric oxide inhibition of human sperm motility.

OBJECTIVE: To determine the effect of nitric oxide (NO) on sperm motility in vitro. DESIGN: Normal human sperm separated by centrifugation through a discontinuous Percoll gradient and subsequent swim-up were incubated for up to 24 hours with NO donors, with and without the known NO quencher hemoglobin, as well as with agents that raise intracellular cyclic 3',5'-guanosine monophosphate (cGMP). Sperm respiration was determined by a tetrazolium-formazan spectrophotometric assay. SETTING: Andrology laboratory. MAIN OUTCOME MEASURES: Absolute sperm motility and respiration. RESULTS: Sperm incubated with the NO donors 1 mM nitroprusside, 100 to 125 microM 3-morpholinosydnonimine, and 25 to 125 microM pure nitric oxide gas dissolved in buffer were inhibited in motility in a dose-dependent fashion. The inhibition could be reversed by the NO quencher hemoglobin. Agents that raise cellular cGMP (dibutyryl cGMP or 8-bromo-cGMP) did not inhibit motility. Nitric oxide inhibited sperm respiration, as measured by the tetrazolium-formazan assay. CONCLUSIONS: Nitric oxide reduces sperm motility, possibly by a mechanism involving inhibition of cellular respiration independent of an elevation of intracellular cGMP. Nitric oxide elaborated in the female or male genital tract in vivo could adversely influence sperm function and fertility.

Cyclic GMP↗

Dietary intervention study to assess estrogenicity of dietary soy among postmenopausal women.

We tested the hypothesis that postmenopausal women on a soy-supplemented diet show estrogenic responses. Ninety-seven postmenopausal women were randomized to either a group that was provided with soy foods for 4 weeks or a control group that was instructed to eat as usual. Changes in urinary isoflavone concentrations served as a measure of compliance and phytoestrogen dose. Changes in serum FSH, LH, sex hormone binding globulin, and vaginal cytology were measured to assess estrogenic response. The percentage of vaginal superficial cells (indicative of estrogenicity) increased for 19% of those eating the diet compared with 8% of controls (P = 0.06 when tested by ordinal logistic regression). FSH and LH did not decrease significantly with dietary supplementation as hypothesized, nor did sex hormone binding globulin increase. Little change occurred in endogenous estradiol concentration or body weight during the diet. Women with large increases in urinary isoflavone concentrations were not more likely to show estrogenic responses than were women with more modest increases. On the basis of published estimates of phytoestrogen potency, a 4-week, soy-supplemented diet was expected to have estrogenic effects on the liver and pituitary in postmenopausal women, but estrogenic effects were not seen. At most, there was a small estrogenic effect on vaginal cytology.

Aged↗

Expanded polytetrafluoroethylene (Gore-Tex Surgical Membrane) is superior to oxidized regenerated cellulose (Interceed TC7+) in preventing adhesions.

OBJECTIVE: To compare the impact of expanded polytetrafluoroethylene (PTFE; Gore-Tex Surgical Membrane; W. L. Gore & Associates, Inc., Flagstaff, AZ) and oxidized regenerated cellulose (Interceed TC7, Johnson & Johnson Medical, Inc., Arlington, TX) on the development of postsurgical adhesions. DESIGN: A multicenter, nonblinded, randomized clinical trial. SETTING: University medical centers. INTERVENTIONS: Each barrier was allocated randomly to the left or right sidewall of every patient. PATIENTS: Thirty-two women with bilateral pelvic sidewall adhesions undergoing reconstructive surgery and second-look laparoscopy. MAIN OUTCOME MEASURES: Adhesion score (on a 0- to 11-point scale), the area of adhesion (cm2), and the likelihood of no adhesions. RESULTS: The use of both barriers was associated with a lower adhesion score and area of adhesion postoperatively. However, those sidewalls covered with PTFE had a significantly lower adhesion score (0.97 +/- 0.30 versus 4.76 +/- 0.61 points, mean +/- SEM) and area of adhesion (0.95 +/- 0.35 versus 3.25 +/- 0.62 cm2). Overall, more sidewalls covered with PTFE had no adhesions (21 versus 7) and, when adhesions were present on the contralateral sidewall, the number of sidewalls covered with PTFE without adhesions was greater than those covered with oxidized regenerated cellulose (16 versus 2). CONCLUSION: Expanded polytetrafluoroethylene was associated with fewer postsurgical adhesions to the pelvic sidewall than oxidized regenerated cellulose.

Adolescent↗

The formation of coalescing peritoneal adhesions requires injury to both contacting peritoneal surfaces.

OBJECTIVE: To determine whether surgical trauma to one or both contacting peritoneal surfaces is necessary to cause coalescing adhesions. SETTING: Research laboratory. DESIGN: The abdominal wall peritoneum and one or both contacting medial peritoneal surfaces of surgically approximated uterine horns in mice were injured by electrocautery, cutting, scratching, or scraping. Adhesion formation was assessed visually and histologically 3 and 7 days later. RESULTS: Regardless of the type of peritoneal injury, few adhesions resulted when only a single injury was made to the abdominal wall (< or = 6%) or to one uterine horn (< or = 13%). When both opposing uterine surfaces were injured, however, adhesions formed at 57% of the sites after electrocautery, 100% after cutting, 100% after scratching, but 0% after scraping. When previously created uterine adhesions were lysed, they reformed at 15 of 15 sites with and 12 of 13 (92%) sites without electrocautery for hemostasis at the time of lysis. CONCLUSIONS: In this murine model, the development of postsurgical adhesions required surgical trauma to both contacting peritoneal sites, regardless of the type of injury, the mobility of the opposing peritoneal surfaces or whether hemostasis was achieved. The clinical implications are that more attention needs to be focused on protecting contacting normal peritoneal surfaces from inadvertent injury during surgery and that different therapeutic strategies may be required for prevention of adhesion formation and reformation because of the high probability of contact between injured peritoneal surfaces with the latter.

Abdominal Muscles↗

An elicited intraperitoneal inflammatory response has no effect on the establishment of pregnancy in the mouse.

OBJECTIVE: To determine the impact of intraperitoneal inflammation on reproduction in the mouse. DESIGN: The effect of an elicited sterile intraperitoneal inflammatory exudate and the passive intraperitoneal transfer of activated syngeneic leukocytes on mating efficiency and uterine implantations was evaluated in mice. SETTING: Research laboratory. INTERVENTIONS: Intraperitoneal injection of thioglycolate was used to elicit large numbers of activated peritoneal macrophages (mean 24.4 x 10(6) leukocytes/animal) in female CD-1 mice. The impact of this intraperitoneal exudate on mating efficiency and number of uterine horn implantations after gonadotropin-stimulated ovulation was determined. In separate experiments, the ovarian bursa present in this species was opened surgically to provide direct access of peritoneal constituents to the genital tract and the experiments repeated. Identical endpoints were evaluated in a third group of experiments using C3H/HEN syngeneic mice after passive transfer of 2, 5, and 10 x 10(6) similarly activated syngeneic peritoneal leukocytes. RESULTS: Neither the elicitation of a peritoneal inflammatory exudate nor the passive transfer of up to 10 x 10(6) activated syngeneic peritoneal macrophages reduced the mating efficiency or the number of uterine implantations. Furthermore, surgically opening the ovarian bursa did not alter these results, although it was associated with anatomic distortion and lowered the number of implantations in all groups. CONCLUSIONS: We could not confirm the previously published reports suggesting a profound adverse impact of intraperitoneal inflammation on reproduction in mice, even when providing direct continuity between the peritoneal cavity and the genital tract. Consequently, the usefulness of this model needs to be re-evaluated before considering it an adequate paradigm for evaluating potential mechanisms of infertility in women.

Adnexa Uteri↗

Does recommending timed intercourse really help the infertile couple?

Timed sexual intercourse is a frequently prescribed component in the treatment for infertile couples. This recommendation is based on a combination of intuition and data from studies often lacking in methodology. With increasingly sophisticated and expensive methods available to time coitus with the presumptive evidence of ovulation, such as the urinary LH kits, the already significant stress of timed intercourse is compounded by the expense of these timing modalities. There is a complete lack of data demonstrating an increased chance of pregnancy with use of such devices. Yet there is evidence that the stress of timed intercourse is a major problem for infertile couples and may even hinder normal reproductive functioning. Available data suggest that much of the period of peak fertility during the menstrual cycle is missed if coitus is timed with the menstrual calendar, basal body temperature thermometer, or LH kit. In addition, the recommendation of a coital frequency of at least a couple of times per week would not only help reduce stress, but also ensure coitus during the period of the menstrual cycle with the greatest chance of resulting in a pregnancy.

Adult↗

Serum and ascitic fluid levels of interleukin-1, interleukin-6, and tumor necrosis factor-alpha in patients with ovarian epithelial cancer.

BACKGROUND: Recent studies have shown that multiple cytokines are secreted by ovarian epithelial cancer cells. Previous studies have shown that the cancer cell lines secrete macrophage colony-stimulating factor (M-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-1 (IL-1), interleukin-6 (IL-6), and transforming growth factor-alpha (TGF-alpha). Concomitantly, the serum levels of one of the growth factors (M-CSF) was found to be significantly elevated in patients with primary ovarian cancer and in second-look patients. The authors evaluated the serum levels of IL-1 alpha, IL-1 beta, IL-6, and tumor necrosis factor-alpha (TNF-alpha) in patients with primary ovarian epithelial cancer. These levels were then compared with cytokine concentration found in normal peritoneal fluid. METHODS: Enzyme-linked immunosorbent assay (ELISA) was used to determine the levels of cytokines in normal peritoneal fluid, ascites, and serum. RESULTS: In serum, TNF-alpha and IL-6 were significantly increased in primary ovarian cancer patients when compared with control subjects (P < 0.0001 for both cytokines). TNF-alpha and IL-6 were also significantly higher than the levels found in second-look patients (P < 0.007 for TNF-alpha, and P = 0.0002 for IL-6). The levels of IL-1 alpha and beta were not elevated in ovarian cancer. TNF-alpha in the ascites was higher when compared with normal peritoneal fluid and was statistically significantly different when a cut-off point between 71-110 pg was selected (P < 0.005). The levels of IL-6 in ascites from patients with primary ovarian cancer also showed a marked increase (P < 0.0001) when compared with peritoneal fluid from control subjects. CONCLUSIONS: Levels of IL-1, IL-6, and TNF-alpha were determined in normal peritoneal fluid, ovarian malignant ascites, normal serum, and serum from patients with ovarian cancer. This study showed that the patients with ovarian cancer have elevated levels of IL-6 and TNF-alpha in serum and ascitic fluid. A larger study would help in evaluating the potential use of cytokines as tumor markers in ovarian cancer.

Ascitic Fluid↗