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Cells actively engaged in de novo steroidogenesis rely on an expansive intracellular network to efficiently transport cholesterol. The final link in the transport chain is STARD1, which transfers cholesterol to the enzyme complex that initiates steroidogenesis. However, the regulation of ovarian STARD1 is not fully characterized, and even less is known about the upstream cytosolic cholesterol transporters STARD4 and STARD6. Here, we identified both STARD4 and STARD6 mRNAs in the human ovary but only detected STARD4 protein since the primary STARD6 transcript turned out to be a splice variant. Corpora lutea contained the highest levels of STARD4 and STARD1 mRNA and STARD1 protein, while STARD4 protein was uniformly distributed across ovarian tissues. Cyclic AMP analog (8Br-cAMP) and phorbol ester (PMA) individually increased STARD1 and STARD4 mRNA along with STARD1 protein and its phosphoform in cultured primary human luteinized granulosa cells (hGCs). STARD6 transcripts and STARD4 protein were unresponsive to these stimuli. Combining lower doses of PMA and 8Br-cAMP blunted the 8Br-cAMP stimulation of STARD1 protein. Increasing cholesterol levels by blocking its conversion to steroid with aminoglutethimide or by adding LDL reduced the STARD4 mRNA response to stimuli. Sterol depletion reduced the STARD1 mRNA and protein response to PMA. These data support a possible role for STARD4, but not STARD6, in supplying cholesterol for steroidogenesis in the ovary. We demonstrate for the first time how cAMP, PMA and sterol pathways separately and in combination differentially regulate STARD4, STARD6 and STARD1 mRNA levels, as well as STARD1 and STARD4 protein in human primary ovarian cells.

Purpose: To determine whether a selected set of mRNA biomarkers expressed in individual cumulus granulosa cell (CC) masses show association with oocyte developmental competence, embryo ploidy status, and embryo outcomes., Methods: This prospective observational cohort pilot study assessed levels of mRNA biomarkers in 163 individual CC samples from 15 women stimulated in antagonist cycles. Nineteen mRNA biomarker levels were measured by real-time PCR and related to the development of their corresponding individually cultured oocytes and subsequent embryos, embryo ploidy status, and live birth outcomes., Results: PAPPA mRNA levels were significantly higher in CC from oocytes that led to euploid embryos resulting in live births and aneuploid embryos compared to immature oocytes by ANOVA. LHCGR mRNA levels were significantly higher in CC of oocytes resulting in embryos associated with live birth compared to immature oocytes and oocytes resulting in arrested embryos by ANOVA. Using a general linearized mixed model to assess ploidy status, CC HSD3B mRNA levels in oocytes producing euploid embryos were significantly lower than other oocyte outcomes, collectively. When transferred euploid embryos outcomes were analyzed by ANOVA, AREG mRNA levels were significantly lower and PAPPA mRNA levels significantly higher in CC from oocytes that produced live births compared to transferred embryos that did not form a pregnancy., Conclusions: Collectively, PAPPA, LHCGR, and AREG mRNA levels in CC may be able to identify oocytes with the best odds of resulting in a live birth, and HSD3B1 mRNA levels may be able to identify oocytes capable of producing euploid embryos.

Purpose: To describe the ultrastructure of spermatozoa from a patient with complete asthenozoospermia that resulted in live births following blastocyst culture., Materials and Methods: Analyses of spermatozoa from a 36 year old patient were performed using light and electron microscopy. The hypo-osmotic swelling test was used to select spermatozoa for intracytoplasmic sperm injection. Embryos were cultured to the blastocyst stage., Results: 100% of the spermatozoa had dynein arm deficiency with secondary defects varying from 3-17%. Six oocytes were injected; five fertilized normally and one was digynic. All five zygotes formed good quality blastocysts. Three blastocysts were cryopreserved and two blastocysts were transferred. Twin females were born at 37 weeks., Conclusions: The hypo-osmotic swelling test can be used to select viable immotile ejaculated spermatozoa from a patient with dynein arm deficiency and can produce excellent fertilization rates and blastocyst development resulting in live births.

Objective: The purpose of this study was to assess whether age, resting follicle number, intraovarian blood flow, and/or ovarian volume are predictive of follicular response and oocyte quality in patients with in vitro fertilization., Study Design: A retrospective chart review was done on 90 in vitro fertilization cycles that were completed in a university setting. Multiple linear regression analysis was used to assess the predictive value of age, resting follicle number, pretreatment ovarian volume, and peak systolic velocity for response to gonadotropin stimulation., Results: Age was predictive (P <.05) for the number of oocytes retrieved, the number that reached two pronuclei, and the number of embryos that reached > or = 4 cells by day 3 after insemination. Resting follicle number was also predictive of follicular response and subsequent embryonic development. Peak systolic velocity and ovarian volume were both found to be of no value., Conclusion: Age and resting follicle number can be used as additional parameters to support patient counseling before in vitro fertilization stimulation.

Background: Approximately 20% of reproductive age couples have difficulty conceiving or maintaining an established pregnancy. The family physician is in a unique position to provide patient education, begin initial evaluation, make appropriate referrals, and offer ongoing counseling and support to couples who experience problems with fertility., Methods: And extensive clinical review was conducted based on a MEDLINE search, the Cochrane database of systematic reviews, and other supporting evidence., Results: Major physiologic influences affecting live birth rates include age, coital frequency, and duration of infertility. Male factor is associated with approximately 40% of these cases and should be addressed early in the evaluation., Conclusion: Many conditions once considered untreatable can now be routinely corrected. As managed care programs expand coverage to include infertility services, primary care providers will be asked to participate in the initial phase of this care. This article offers a practical approach.

Three cases of nonphysiologic hyperprolactinemia associated with pituitary disease evidenced by galactorrhea are presented. Two patients had significant pituitary disease associated with low-level prolactin elevations. The third patient had only a history of infertility and expressible galactorrhea on examination. This patient was found to have high prolactin levels and a locally invasive pituitary tumor. Physicians need to be aware of the serious conditions associated with galactorrhea so that appropriate diagnostic studies can be done and treatment instituted.

Hypersensitivity reactions are a documented complication of urinary-derived gonadotropin therapy. In this report, a patient allergic to these older products is successfully treated with recombinant follicle-stimulating hormone.

Major congenital anomalies occur in approximately 4% of the newborn population. Although this risk is apparently the same for in vitro fertilization (IVF) pregnancies, couples having assisted reproduction therapy should receive appropriate genetic counseling, because coincidental maloccurrences are often erroneously linked to the technology. We report a case of newborn esophageal atresia with tracheoesophageal fistula occurring in an IVF pregnancy. Physicians are encouraged to educate all prospective parents regarding the rate of congenital anomalies in the general population so that they have realistic expectations concerning newborn outcome.

The importance of Rhodococcus equi infection, an uncommon human pathogen that almost exclusively affects immunocompromised hosts, has greatly increased following the advent of acquired immune deficiency syndrome (AIDS) epidemics. Until the present time, 38 cases of R. equi infection have been described in human immunodeficiency virus (HIV)-infected patients; we now report a further five personal cases. R. equi was acquired via respiratory exposure to animals in less than half of the patients, and caused invasive pulmonary infection (91%), bacteraemia and sometimes bloodstream dissemination. R. equi was easily cultured from sputum or blood, but its diagnosis was often difficult due to microbiological and clinical similarities with other pathogens. The persistence of the micro-organism inside macrophages and its high tissular load represent the major limitation to an effective treatment. Several antibiotics are active in vitro, but their efficacy in vivo depends on macrophage uptake and/or bactericidal activity. Treatment should start with at least two intravenous bactericidal antibiotics for 3-4 weeks, and then continue with oral therapy for a period of up to several months with at least two intracellularly active drugs. Surgical resection of the lesions may be beneficial in selected cases.

This review concerns studies of the comparative efficacy and safety of torasemide and furosemide in patients with cirrhosis of the liver complicated by ascites and oedema. The short-term trials reviewed indicated that in patients who had failed to respond with adequate diuresis and loss of body weight and ascites to bed rest, restricted salt and water intake and spironolactone, torasemide had a longer duration of action than furosemide and resulted in a greater urinary excretion of salt and water and greater loss of body weight. Torasemide also had less effect than furosemide on urinary potassium excretion and unlike furosemide did not increase the fractional excretion of magnesium or phosphate or the blood ammonia concentration. Two longer term trials in similar patients with decompensated hepatic cirrhosis confirm the results of the shorter term studies. These studies, albeit each in relatively small numbers of patients, confirm the ability of torasemide to enhance diuresis, free water clearance and fractional excretion of sodium and chloride, resulting in loss of body weight and mobilization of ascites in patients with decompensated hepatic cirrhosis. In these patients, the relatively small increase in urinary excretion of potassium, induced by torasemide without any marked effect on renal function or on the plasma neurohormonal profile, enhances its potential safety.

Two patients seropositive for human immunodeficiency virus (HIV) and with no previous acquired immunodeficiency syndrome-defining conditions developed cavitary pneumonia and pleural disease caused by Rhodococcus equi. R. equi was isolated from these patients' sputum and lung biopsy specimens, respectively, but the microorganism was initially considered to be a contaminant (patient 1) or misidentified as a nontuberculous mycobacterium (patient 2). The R. equi infection was fatal in one patient, who died after 4 months without specific antimicrobial therapy; the second patient was unresponsive to combination therapy with various antimicrobial agents. R. equi may cause life-threatening infections in HIV-infected patients. Microbiology laboratories should be cognizant of the need to exclude R. equi as a cause of infection in highly immunosuppressed patients.

A total of 210 patients consecutively submitted to heart surgery at the Parma University Hospital and transfused with 1,898 units of blood were followed after transfusion in order to evaluate both the incidence of anti-hepatitis C virus (HCV) seroconversion in non-A, non-B post-transfusion hepatitis (PTH-NANB) cases and the usefulness of the screening for anti-HCV in comparison with that for serum glutamic pyruvic transaminase (SGPT) values in preventing PTH-NANB transmission. Fifteen recipients developed PTH-NANB (group A); ten of them (66.6%) showed anti-HCV seroconversion within 3-12 months. Eight of the ten anti-HCV positive patients developed chronic hepatitis, but none of the five PTH-NANB anti-HCV negative did. None of the 15 controls (group B) randomly chosen among the patient population showed anti-HCV seroconversion. A close correlation with the transmission of PTH was showed by anti-HCV positivity but not by SGPT elevation in blood donors. Eleven of 172 blood products transfused to group A but none of 139 products transfused to group B were anti-HCV positive. The incidence of elevated SGPT values was similar between the two groups of the transfused blood products. Nevertheless, the correlation observed between anti-HCV positivity and SGPT levels in the blood products involved in PTH confirms the need to exclude blood donors with abnormal SGPT values. On the whole, anti-HCV screening of donors showed a predictive value higher than that of SGPT (100% vs. 53.3%), allowing a minor blood donation exclusion.(ABSTRACT TRUNCATED AT 250 WORDS)

An epidemiological study of stone disease in a Northern Italian city was carried out by means of a postal questionnaire mailed to 6000 individuals (2.5% of the entire population). It was found that the incidence of stone disease was comparable to that of industrialised Western Europe. There was a relationship between stone disease and gout and stone disease and a positive family history. The frequency of uric acid stones was high (26.5%). Stone-formers showed no alimentary differences from non-stone formers apart from the use of spices and herbs. Stone-formers used less water from public aqueducts and more uncarbonated mineral water, but only 19% of these drank at least 2 litres a day.

A method is described for the determination of oxalate in urine using flow injection analysis and fluorimetry. Oxalate is precipitated with calcium chloride at pH 4.5, redissolved in H2SO4 and measured by flow injection analysis. The minimum detection limit is 6 mumol/l. The coefficient of variation is 7%. Results are in good accordance with normal values found with traditional oxalate analysis.

Insulin-dependent diabetes mellitus (IDDM) induces plasma amino acid (AA) abnormalities, including low alanine and high branched-chain (BCAA). While insulin treatment restores plasma AA pattern, proline, methionine, valine, isoleucine, and total BCAA remain elevated in skeletal muscle intracellular water. This suggests that the restoration of plasma AA concentrations is not a satisfactory index of recovered AA metabolism in IDDM.

The effects of a moderate surgical trauma on body composition, in conditions comparable to fasting were assessed in a balance study. Two groups of patients under normal nutritional conditions were compared after cholecystectomy. The patients underwent for three days the following schedules of parenteral therapy: incomplete nutrition with balanced intake of water, sodium and potassium but carbohydrate deficient and lacking in amino acids (group 1, 10 cases); or complete nutrition with respect to calories, amino acids and other electrolytes (group 2, 9 cases). An evident loss in body weight was observed in patients undergoing incomplete nutrition, not justified by a moderately negative water balance. Statistical comparison of the two groups showed equal or positive balances for group 1 while group 2 gave negative results for all the parameters, particularly for nitrogen and potassium, suggestive of a loss in lean body mass greater than that usually occurring after simple fasting. Results confirmed the poor significance of serum values in expressing even significant degrees of electrolyte depletion. Substantial modifications in body composition may appear as clinical manifestations in cases with pre-existing malnutrition or when drugs that affect or are affected by the water and electrolyte balance are administered.

In patients with renal colic we studied lithogenic urinary risk factors before and after the stone passage. We showed abnormalities in water, electrolytes and other substances excretion due to retention and metabolic disorders. The effects more pronounced is on urinary sodium, calcium, magnesium and ammonium. Citrate behaviour suggests a transient intracellular acidosis.

The question whether body electrolyte composition in patients with primary hypertension differs from that of normotensive subjects is still controversial. The aim of the present work was to investigate on water, electrolyte, and nitrogen muscle cell composition in essential human hypertension. Also the effects of antihypertensive drugs on muscle electrolyte contents were analyzed. The results indicate that muscle Na was higher and muscle K was lower in essential hypertensives in comparison with controls. Muscle water, Mg, and nitrogen were not different in treated patients (with or without diuretics) in comparison with untreated hypertensive patients. But in hypertensive patients treated with diuretics, Nam was higher and Km was lower than in untreated patients. Several mechanisms may explain the changes on muscle electrolyte composition in primary hypertension, but the cellular mechanism involving abnormalities in ion pump and electrolyte transmembrane fluxes are the most probable. The therapy with antihypertensive drugs especially diuretics, seems to enhance different muscle electrolyte patterns in essential hypertension.