Effect of metformin and flutamide on insulin, lipogenic and androgen-estrogen signaling, and cardiometabolic risk in a PCOS-prone metabolic syndrome rodent model.

Polycystic ovary syndrome (PCOS) is highly associated with cardiometabolic risk and the metabolic syndrome (MetS), predisposing women to increased risk of developing type 2 diabetes and cardiovascular disease. Metformin is commonly used to treat insulin resistance-glucose intolerance, and flutamide, an androgen receptor (AR) antagonist, is used to target hyperandrogenemia and dyslipidemia. Currently, the physiological mechanism of action of these treatments on androgen, lipidogenic, and insulin signaling pathways remains unclear in PCOS. The aim of this study was to investigate the effects and mechanisms of action of metformin and flutamide on plasma lipid-apolipoprotein (Apo)B-lipoprotein and insulin-glucose metabolism, and endocrine-reproductive indices in a PCOS-prone MetS rodent model. PCOS-prone rodents were treated with metformin (300 mg/kg body wt), flutamide (30 mg/kg body wt), or metformin + flutamide combination treatment for 6 wk. Metformin was shown to improve fasting insulin and HOMA-IR, whereas flutamide and combination treatment were shown to reduce plasma triglycerides, ApoB48, and ApoB100, and this was associated with decreased intestinal secretion of ApoB48/triglyceride. Flutamide and metformin were shown to reduce plasma androgen indices and to improve ovarian primary and preovulatory follicle frequency. Metformin treatment increased hepatic estrogen receptor (ER)α, and metformin-flutamide decreased intestinal AR and increased ERα mRNA expression. Metformin-flutamide treatment upregulated hepatic and intestinal insulin signaling, including insulin receptor, MAPK1, and AKT2. In conclusion, cardiometabolic risk factors, in particular ApoB-hypertriglyceridemia, are independently modulated via the AR, and understanding the contribution of AR and insulin-signaling pathways further may facilitate the development of targeted interventions in high-risk women with PCOS and MetS.

Structure of the oligogalacturonate-specific KdgM porin.

The phytopathogenic Gram-negative bacterium Dickeya dadantii (Erwinia chrysanthemi) feeds on plant cell walls by secreting pectinases and utilizing the oligogalacturanate products. An outer membrane porin, KdgM, is indispensable for the uptake of these acidic oligosaccharides. Here, the crystal structure of KdgM determined to 1.9 Šresolution is presented. KdgM is folded into a regular 12-stranded antiparallel ß-barrel with a circular cross-section defining a transmembrane pore with a minimal radius of 3.1 Å. Most of the loops that would face the cell exterior in vivo are disordered, but nevertheless mediate contact between densely packed membrane-like layers in the crystal. The channel is lined by two tracks of arginine residues facing each other across the pore, a feature that is conserved within the KdgM family and is likely to facilitate the diffusion of acidic oligosaccharides.

Utilization of 2-monoacylglycerols for phosphatidylcholine biosynthesis in the intestine.

Conventional preparations of intestinal microsomes were observed to incorporate acetone-solubilized 2-oleoyl-[2-3H]glycerol into dioleoylglycerophosphocholine in the presence of oleoyl CoA and CDP-choline. The apparent Km values for CDP-choline utilization were 77 +/- 10 microM in rat and 72 +/- 5 microM in hamster. The incorporation ratio of glycerol into triacylglycerols and phosphatidylcholines was 4.5:1 and 25:1 in the rat and hamster, respectively. Endogenous diacylglycerols generated by phospholipase C treatment of microsomes readily equilibrated with the diacylglycerols arising via the monoacylglycerol pathway as indicated by a dilution of the radioactivity in the triacylglycerol and phosphatidylcholine synthesized from radioactive 2-monooleoylglycerol. These results suggest an alternative pathway for glycerophospholipid formation in the intestinal mucosa during possible inhibition of the phosphatidic acid pathway by dietary 2-monoacylglycerols. It is concluded that exogenously added monoacylglycerol can serve as a precursor for microsomal diacyl- and triacylglycerol as well as phosphatidylcholine. The inability to demonstrate comparable monoacylglycerol utilization in earlier experiments in attributed to the inhibition of choline phosphotransferase by the detergents used to solubilize the acylglycerols.

The cloning and expression of a murine triacylglycerol hydrolase cDNA and the structure of its corresponding gene.

A novel murine cDNA for triacylglycerol hydrolase (TGH), an enzyme that is involved in mobilization of triacylglycerol from storage pools in hepatocytes, has been cloned and expressed. The cDNA consists of 1962 bp with an open reading frame of 1695 bp that encodes a protein of 565 amino acids. Murine TGH is a member of the CES1A class of carboxylesterases and shows a significant degree of identity to other carboxylesterases from rat, monkey and human. Expression of the cDNA in McArdle RH7777 hepatoma cells showed a 3-fold increase in the hydrolysis of p-nitrophenyl laurate compared to vector-transfected cells. The highest expression of TGH was observed in the livers of mice, with lower expression in kidney, heart, adipose and intestinal (duodenum/jejunum) tissues. The murine gene that encodes TGH was cloned and exon-intron boundaries were determined. The gene spans approx. 35 kb and contains 14 exons. The results will permit future studies on the function of this gene via gene-targeting experiments and analysis of transcriptional regulation of the TGH gene.

Expression of survivin in normal, hyperplastic, and neoplastic colonic mucosa.

The regulation of apoptotic cell death may have a profound effect on the pathogenesis and progression of colon cancer. Survivin, a member of the inhibitor of apoptosis gene family, has been detected in fetal tissue and in a variety of human malignancies. In the current study, we investigated survivin expression by an immunohistochemical approach in benign, hyperplastic, premalignant, and malignant lesions of the colon. Survivin was detected in all cases of normal colonic mucosa (20/20), hyperplastic polyps (20/20), adenomatous polyps (20/20), and in both well differentiated and moderately differentiated colonic adenocarcinomas (20/20). In the normal colonic mucosa, survivin expression was mostly restricted to the base of the colonic crypts. All epithelial cells showed uniformly intense staining for survivin in hyperplastic polyps. By contrast, adenomas and adenocarcinomas showed a heterogeneous staining pattern with cell-to-cell, gland-to-gland, and regional variability in the intensity of survivin staining. In contrast to the basal preponderance of staining in normal colonic mucosa, numerous survivin positive cells were present at the luminal surface of hyperplastic polyps, adenomatous polyps, and adenocarcinomas. In conclusion, the expression of survivin is not a specific marker of adenocarcinoma of the colon but does show characteristic and reproducible patterns of expression in non-neoplastic proliferative lesions and in normal colonic mucosa.

Cytotoxicity of pivalylphenylephrine and pivalic acid to corneal endothelium.

We examined the effects of topically applied pivalylphenylephrine (PPE) and pivalic acid (PA) on the corneal endothelium of rabbits and the direct effects of PPE and PA on monolayer cultures of bovine corneal endothelium. The PPE-treated corneas without epithelium significantly increased in thickness, whereas no change in thickness was observed in corneas with epithelium intact. The PA did not alter the thickness of corneas with or without epithelium. Although 0.001% PE had no noticeable effect in two hours, 0.01% PPE caused breakdown of intercellular junctions in cultured cells in five minutes. Higher concentrations of PPE caused the cells to detach from the culture dishes within 30 minutes of treatment. Only 1% PA caused cell elongation and loss of intercellular contact after 60 to 90 minutes of exposure; lower concentrations did not effect cultured cells.

Influence of delayed staging laparotomy after laparoscopic removal of ovarian masses later found malignant.

OBJECTIVE: To determine whether delayed laparotomy after attempted laparoscopic excision of an ovarian mass later found to be malignant has an impact on the stage of disease. METHODS: A questionnaire regarding laparoscopic management of ovarian masses later found to be malignant was mailed to all gynecologic departments in Austria. Of the 70 cases reported, laparotomy was performed after laparoscopy in 48 cases. In 24 of these cases, laparotomy was performed within 17 days of laparoscopy, whereas 24 cases involved a delay of more than 17 days. Twenty-two patients in whom laparotomy was performed immediately after laparoscopy were used as controls. RESULTS: In patients with borderline tumors who underwent laparotomy more than 17 days after laparoscopy, the odds ratio (OR) for International Federation of Gynecology and Obstetrics (FIGO) stage IIB-IV disease was 5.3 (95% confidence interval [CI] 0.40, infinity), compared with patients undergoing immediate laparotomy (multivariate analysis). Patients with invasive ovarian cancer who underwent laparotomy more than 17 days after laparoscopy had an OR of 9.2 (CI 0.92, 481) for stage IIB-IV disease compared with patients undergoing immediate laparotomy (multivariate analysis). In patients with borderline tumors, multivariate analysis showed that the timing of laparotomy is an independent prognostic factor for the stage of disease. In invasive ovarian cancer, none of the factors evaluated by multivariate analysis was found to be an independent prognostic factor for the distribution of disease stage. A delay between laparoscopy and laparotomy may affect adversely the distribution of disease stage. CONCLUSION: The timing of subsequent laparotomy was found to be a factor predictive of the distribution of disease stage.

Localization of telomerase hTERT protein and survivin in placenta: relation to placental development and hydatidiform mole.

OBJECTIVE: To find if a difference in telomerase or survivin expression exists between non-neoplastic tissues and hydatidiform moles, and explore expression of those proteins in normal placental development, post-term gestation, and preeclampsia. METHODS: Formalin-fixed placental tissues were selected from collections of the Department of Pathology at the University of Colorado. Five specimens of each trimester, five each of preeclamptic and post-term placentas, and 23 molar pregnancies were selected. The telomerase catalytic protein hTERT was localized in placental tissues using the catalyzed signal amplification system, and survivin was localized by conventional immunoperoxidase method. Staining was graded on a scale of zero to 4. RESULTS: hTERT staining was detected in sections of 42 of 48 specimens (23 of 23 hydatidiform moles, 19 of 25 non-neoplastic placental tissues). The intensity of staining for hTERT was higher in hydatidiform moles (mean 3.3, median 3) compared with levels in non-neoplastic placental tissues (mean 0.92, median 1) (P <.001). Survivin was detected in 39 of 48 specimens (22 of 23 hydatidiform moles, 17 of 25 non-neoplastic placental tissues). Compared with non-neoplastic tissues (mean 0.88, median 1), survivin levels were elevated in hydatidiform moles (mean 1.35, median 1) (P =.031). CONCLUSION: Survivin and telomerase were increased in hydatidiform moles, suggesting that regulation of apoptosis and stabilization of telomere length might be involved in neoplastic transformation of the placenta. The patterns of expression observed for survivin and telomerase in non-neoplastic placental tissues suggest that the control of apoptosis and stabilization of telomeric DNA might also be involved in normal gestational development.

Fallopian tissue sampling with a cytobrush during hysteroscopy: a new approach for detecting tubal infection.

OBJECTIVE: To evaluate whether the transcervical approach for fallopian tissue sampling is a practicable and safe method to establish an etiologic diagnosis of salpingitis. DESIGN: Controlled clinical study. SETTING: Academic research environment. PATIENT(S): Twenty women with primary or secondary sterility undergoing hysteroscopy and laparoscopy. INTERVENTION(S): During hysteroscopy, fallopian tissue samples were obtained from the proximal parts of both tubes with a cytobrush inserted through the working channel of the hysteroscope. Diagnostic laparoscopy with assessment of tubal patency was performed in the same session. The specimens were split and used for chlamydial testing by both cell culture and polymerase chain reaction. Serum samples were evaluated for the presence of antichlamydial antibodies. MAIN OUTCOME MEASURE(S): Practicability and safety of the method. RESULT(S): The sampling procedure was performed in all patients without complications. Adequate samples were obtained from both tubes in all 20 patients. The fallopian specimens showed evidence of chlamydial infection in one patient. CONCLUSION(S): Those data indicate that transcervical fallopian tissue sampling with a cytobrush is a reliable and safe technique.

Successful treatment of a heterotopic pregnancy by sonographically guided instillation of hyperosmolar glucose.

OBJECTIVE: To describe a new conservative approach in the treatment of heterotopic pregnancy. DESIGN: Case report. SETTING: A university hospital and an outpatient IVF center. INTERVENTION(S): Transvaginal ultrasonography-guided puncture of the ectopic gestational sac and instillation of hyperosmolar glucose. RESULT(S): The ectopic pregnancy resolved without further intervention. The intrauterine pregnancy resulted in a full-term live birth. CONCLUSION(S): Transvaginal injection of hyperosmolar glucose may be an effective conservative treatment for unruptured ectopic pregnancies in cases of heterotopic pregnancy.

Phonocardiogram signal analysis: a review.

Many disease of the heart cause changes in heart sounds and additional murmurs before other signs and symptoms appear. Hence, heart sound analysis by auscultation is the primary test conducted by physicians to assess the condition of the heart. Yet, heart sound analysis by auscultation as well as analysis of the phonocardiogram (PCG) signal have not gained widespread acceptance. This is due mainly to many controversies regarding the genesis of the sounds and the lack of quantitative techniques for reliable analysis of the signal features. The heart sound signal has much more information than can be assessed by the human ear or by visual inspection of the signal tracings on paper as currently practiced. Here, we review the nature of the heart sound signal and the various signal-processing techniques that have been applied to PCG analysis. Some new research directions are also outlined.

Survivin expression in human osteosarcoma is a marker for survival.

AIMS: Osteosarcoma is the most frequent malignant bone tumor with a peak incidence in the second and third decade of life. Evaluation of prognosis of patients with osteosarcoma is limited to clinical parameters whereas molecular markers of tumor aggression have not yet been identified. Inhibition of apoptotic cell death could play a role in the development or progression of neoplasia. Survivin is a member of the inhibitor of apoptosis (IAP) protein gene family and is expressed both during normal fetal development and in human cancer. METHODS: The localization and distribution of survivin was investigated immunohistochemically in high-grade osteosarcomas by an indirect immunoperoxidase method. RESULTS: Survivin was detected in the cytoplasm in 23/40 and in the nucleus in 20/40 cases of osteosarcoma. Nuclear localization of survivin expression was significantly correlated with a prolonged survival (P=0.0347) but cytoplasmic staining showed no correlation with patient outcome. CONCLUSIONS: The results of this study indicates that the evaluation of survivin expression might be a useful prognostic marker in osteosarcoma. Patients with osteosarcoma exhibiting nuclear survivin expression could potentially benefit from stratification of neoadjuvant chemotherapy.

Acoustomechanical properties of open TTP titanium middle ear prostheses.

OBJECTIVE: The purpose of the study was to identify acoustcomechanical properties of various biostable and biocompatible materials to create a middle ear prosthesis with the following properties: (i) improved handling including a good view of the head of the stapes or footplate and adjustable length, (ii) improved acoustical characteristics that are adequate for ossiculoplastic. The identified material should serve to build CE and FDA approved prostheses for clinical use in patients. METHODS: Test models made of Teflon, polyetheretherketone, polyethylenterephtalate, polysulfone, gold, Al2O3 ceramics, carbon and titanium were investigated for their potential to fulfill the requirements. Acoustical properties were investigated by laser Doppler velocimetry (LDV) in mechanical middle ear models (MMM). Measured data were fed in to a recently created computer model of the middle ear (multibody systems approach, MBS). Using computer-aided design (CAD) measured and computed data allowed creation and fine precision of titanium prostheses (Tübingen Titanium Protheses, TTP). Their handling was tested in temporal bones. Acoustomechanical properties were investigated using the MBS and mechanical middle ear models. MAIN OUTCOME MEASURES: Input impedance, mass, stiffness, and geometry of test models and prostheses were determined. Furthermore, their influence on the intraprosthetic transfer functions and on coupling to either tympanic membrane or stapes was investigated. RESULTS: Final results were FDA- and CE-approved filigreed titanium prostheses with an open head that fulfilled the four requirements detailed above. The prostheses (TTP) were developed in defined lengths of between 1.75 and 3.5 mm (partial) and 3.0 and 6.5 mm (total) as well as in adjustable lengths (TTP-Vario). CONCLUSIONS: The results suggest acoustomechanical advantages of TTPs because they combine a significantly low mass with high stiffness. In contrast to closed prostheses, the open head and filigreed design allow an excellent view of the prosthesis foot during coupling to the head or footplate of stapes, contributing to an improved intraoperative reliability of prosthesis coupling.

Contraceptive knowledge and attitudes of Austrian adolescents after mass media reports linking third-generation oral contraceptives with an increased risk of venous thromboembolism.

We performed a representative survey to determine the level of knowledge of 1,010 Austrian adolescents aged 14 to 24 years about selected facts relating to the recent massive news coverage of the increase in the risk of venous thromboembolism in users of third-generation oral contraceptives and to assess the contraceptive behavior of this population. The overall use rate of oral contraceptives and condoms had increased significantly between 1991 and 1996. Sixty-six percent of the adolescents surveyed stated not having heard or read any media reports on oral contraceptives. Only 8% of those who had knew that most reports focused on the pill as a possible cause of venous thromboembolism, whereas the majority of respondents indicated that the media conveyed doubts regarding the health safety of oral contraceptives in general. Nearly half of adolescents were unable to define what a thrombosis was. Thus, although the mass media play an important role in transmitting medical information, the dissemination of practical, accurate advice on the risks of a drug and competent patient counseling is reserved for the health care professionals.

Stereospecificity of monoacylglycerol and diacylglycerol acyltransferases from rat intestine as determined by chiral phase high-performance liquid chromatography.

Using chiral phase high-performance liquid chromatography of diacylglycerols, we have redetermined the ratios of 1,2-/2,3-diacyl-sn-glycerols resulting from acylation of 2-monoacylglycerols by membrane bound and solubilized triacylglycerol synthetase of rat intestinal mucosa. With 2-oleoyl[-3H]glycerol as the acyl acceptor and oleoyl-CoA as the acyl donor, 97-98% of the diacylglycerol product was 1,2(2,3)-dioleoyl-sn-glycerol, 90% of which was the sn-1,2- and 10% the sn-2,3-enantiomer. The remaining diacylglycerol (less than 3%) was the sn-1,3-isomer. The overall yield of acylation products was 70%, of which 60% were diacylglycerols and 40% triacylglycerols. With 2-oleylglycerol ether as the acyl acceptor and [1-14C]oleoyl-CoA as the acyl donor, 90% of the diradylglycerol was 1-oleoyl-2-oleyl-sn-glycerol and 10% was the 2-oleyl-3-oleoyl-sn-glycerol. The diradylglycerols made up 96% and the triradylglycerols 4% of the radioactive product. With 1-palmitoyl-sn-glycerol as the acyl acceptor and [1-14C]oleoyl-CoA as the acyl donor, the predominant reaction product was 1-palmitoyl-3-oleoyl-sn-glycerol. The 3-palmitoyl-sn-glycerol was not a suitable acyl acceptor. Both 1,2- and 2,3-diacyl-sn-glycerols were substrates for diacylglycerol acyltransferase as neither isomer was favored when 1,2-dioleoyl-rac-[2-3H]glycerol was used as the acyl acceptor. There was a marked decrease in the acylation of the 1(3)-oleoyl-2-oleyl-sn-glycerol to the 1,3-dioleoyl-2-oleyl-sn-glycerol. It is concluded that neither monoacylglycerol nor diacylglycerol acyltransferase exhibit absolute stereospecificity for acylglycerols as fatty acid acceptors.

The diagnostic value of planar and SPET scintimammography in different age groups.

Planar scintimammography (SMM) with 99Tcm-sestamibi (99Tcm-MIBI) has proved to be a useful method in the evaluation of breast lesions. The aim of our study was to determine the diagnostic value of planar and single photon emission tomograpic (SPET) SMM in different age group. We investigated women with breast lesions for which the surgeon recommended biopsy or careful follow-up. Seventy consecutive patients underwent supine planar SMM, while supine SPET was performed in 68 of the patients. Twenty-seven carcinomas in 23 patients and 35 benign lesions in 31 patients were confirmed histologically. In the remaining 16 patients, the findings were classified as benign at follow-up. The carcinomas and benign lesions had a mean (+/- S.D.) diameter of 25 +/- 17 and 17 +/- 11 mm respectively (P < 0.05). A sensitivity/specificity of 67/96% and 88/91% was obtained with the planar and SPET techniques respectively. The accuracy of both techniques was 90%. In patients aged 40 years or less, SPET increased the sensitivity from 50 to 100% (P < 0.01), whereas it decreased the specificity and accuracy from 94 to 81% and 91 to 81% respectively (N.S.). In patients aged 41-50 years, the sensitivity increased from 63 to 100% (P < 0.01), whereas there was a small decrease in specificity and an increase in accuracy (from 91 to 89% and 86 to 91% respectively, N.S.) We conclude that, in younger patients, a negative SPET SMM study indicates the need for careful follow-up rather than biopsy. However, the addition of SPET in supine scintimammography does-not change the overall accuracy significantly.

Placental insufficiency and maternal death caused by advanced stage of breast cancer in third trimester.

We describe a case with placental, and general metastases, resulting in transient intrauterine and general hypoxia, and with additionally clinical features similar to HELLP syndrome. A patient in the third trimester with dyspnea at rest developed right heart failure during c-section. During emergency thoracotomy the patient went into generalized shock and died after intense CPR. Placental insufficiency was based on a multilocal metastatic event, decreasing the utero-placental perfusion.

Open Tübingen titanium prostheses for ossiculoplasty: a prospective clinical trial.

OBJECTIVE: The overall purpose of the study was the evaluation of the efficacy of Tübingen titanium prostheses (TTPs) for ossiculoplasty. STUDY DESIGN: A two-part clinical study of 216 patients undergoing ossiculoplasty was performed. The first part was a prospective study using TTPs (n = 114). The second part involved study of historical control patients (n = 102) with gold and ceramic prostheses. INTERVENTIONS: All patients underwent ossiculoplasty. MAIN OUTCOME MEASURES: Measures included median air conduction thresholds and air-bone gaps. RESULTS: All patients were per-protocol patients. When the air-bone gap "gold standard" (i.e., < or =10 dB) was investigated in the main speech spectrum, partial TTPs reached this level at 2 kHz in 44% (n = 22) and at 3 kHz in 38% (n = 19). Gold and ceramics revealed significantly lower values. Similar results were obtained for total prostheses. Differences for TTPs and ceramics were statistically significant (Mann-Whitney U test, alpha = 5%). CONCLUSION: The use of TTPs for ossiculoplasty is an efficient treatment method.

Total implantation of the active hearing implant TICA for middle ear disease: a temporal bone study.

A subpopulation of hearing-impaired patients has conductive hearing loss that cannot be improved by classic tympanoplasty. Other patients have a mixed hearing loss and cannot be helped by present forms of ear surgery or by hearing aids. Possible help for some patients may come from current implantable hearing devices if these are modified for the patient's specific anatomic situation. The TICA LZ 3001 is a hearing implant for total implantation used to treat moderate to severe sensorineural hearing loss. Most patients who use it have a normal ossicular chain that allows coupling of the implant to the incus. The present temporal bone study demonstrates that the TICA can also be used in patients with an interrupted ossicular chain. If the incus long process shows a defect, the TICA may be coupled to the incus body, and connection between the stapes and the long process of the incus can be achieved with a commercially available titanium-angle prosthesis or liquid ionomeric cement. In cases of an absent incus, the coupling axis of the transducer may be coupled to the stapes head via a modified coupling element. With an absent stapes, the coupling axis may be coupled directly to the perilymph by a coupling element similar to a gold stapes prosthesis.

Lectin histochemistry of fallopian tube epithelial cells. Relation to ovum transport and ovum pickup.

OBJECTIVE: Data on histochemical and biochemical characteristics of the human oviduct are scarce. The exact mechanisms of ovum transport and pickup are not fully understood. STUDY DESIGN: Human fallopian tubes were obtained and prepared for histochemistry. We analyzed the distribution of negatively charged groups on the oviduct epithelium and cumulus cells and examined the distribution of glycoconjugates by means of lectin histochemistry. We tested the possible influence of poly-L-lysine and considered ABO blood group expression since these characteristics are determined by specific terminal sugar residues. RESULTS: A negatively charged glycocalyx exists on tubal epithelial cells and cumulus cells. Adherence by affinities similar to sugar-lectin binding forces could be disproven in case of commonly used lectins. Poly-L-lysine inhibited the cationic binding reaction but did not influence lectin binding. The blood group A glycoprotein presents terminal D-N-acetyl-galactosamine residues, which are demonstrated by HPA lectin binding. CONCLUSION: Our study indicates that it is unlikely that electrostatic interactions play a major role in ovum transport or pickup. Since poly-L-lysine has been described as inhibiting ovum transport, sugar-lectin binding affinities seem not to operate in ovum transport or pickup.