Proteomic characterization of the qualitative and quantitative differences in cervical mucus composition during the menstrual cycle.

The chemical composition of the cervical mucus (CM), its physical characteristics and the volume of secretion change cyclically throughout the menstrual cycle. The aim of this study was to identify the constitutive protein composition of CM of fertile women and the changes in the CM proteome throughout the menstrual cycle. Five fertile women who had a term delivery within 1 year before the study were enrolled. Proteomic analysis was performed using an Ultimate 3000 Nano/Micro-HPLC apparatus equipped with an FLM-3000-Flow manager module and coupled with an LTQ Orbitrap XL hybrid mass spectrometer; bioinformatic software was used for functional and quantitative analysis. 59, 81 and 43 proteins (mean) were respectively identified in the pre-ovulatory, ovulatory and post-ovulatory samples. 38 common proteins were identified. 42, 38 and 17 exclusive proteins were respectively identified in pre-ovulatory, ovulatory and post-ovulatory CM. The main part of CM constituents has a catalytic activity, which is mainly related to hydrolase activity. The label-free quantitative analysis of the common proteins revealed a significant reduction in the protein abundance index for antileukoproteinase, after the ovulation, and a peak of haptoglobin at ovulation. This is the first application of high-resolution MS-based proteomics for the identification of protein constituents of CM. This approach may contribute to the identification of putative biomarkers of the female reproductive tract.

Male fertility and reduction in semen parameters: a single tertiary-care center experience.

Background. Infertility is both a clinical and a public problem, affecting the life of the couple, the healthcare services, and social environment. Standard semen analysis is the surrogate measure of male fertility in clinical practice. Objective. To provide information about the relationship between semen parameters and spontaneous conception. Methods. We evaluated retrospectively 453 pregnancies that occurred among 2935 infertile couples evaluated at an infertility clinic of a tertiary-care university hospital, between 2004 and 2009. Results. Normal semen analysis was present only in 158 patients; 295 subfertile patients showed alterations in at least one seminal parameter. A reduction in all seminal parameters was observed in 41 patients. Etiological causes of male infertility were identified in 314 patients. Conclusion. Our data highlights the possibility of a spontaneous conception with semen parameters below WHO reference values. Therefore, we support the importance of defining reference values on a population of fertile men. Finally, we analyzed the related ethical issues.

[Role of a mini-invasive approach in the diagnosis and treatment of tubo-peritoneal infertility as an altenative to IVF]. / Il ruolo di un approccio chirurgico mini-invasivo nello diagnosi e trattamento della sterilità tubo-peritoneale in alternativa alla FIVET.

AIM: Tubal factor infertility accounts for approximately 25-35% of cases of female infertility. Identifiable causes of tubal infertility are postinfectious tubal damage, postsurgical adhesion formation, and endometriosis-related adhesions. Aim of this study was to evaluate the results of a diagnostic/therapeutic minimally invasive approach in patients with suspect or ascertained mechanical infertility in terms of obtained pregnancies. METHODS: The study enrolled 143 patients who underwent diagnostic or operative laparoscopy, with chromopertubation, peritoneal or endometrial culture, salpingoscopy when indicated and diagnostic or operative hysteroscopy. Nine patients with submucous-intramural or multiple intramural fibroids underwent miomectomy by minilaparotomy following hysteroscopy and chromopertubation. Patients were contacted periodically by telephone to monitor the onset and outcome of pregnancy. The mean length of follow- up was 49 months (range: 11 to 118 months). RESULTS: Of the 152 patients considered in the study, 61 became pregnant (40%). Twenty-three pregnancies resulted in miscarriage, two in tubal pregnancy and one patient aborted after a diagnosis of Down syndrome. In total, 32% of the patients achieved a term pregnancy. CONCLUSION: The diagnostic/therapeutic mini-invasive approach allows women to become pregnant naturally and it is, therefore, an option for couples with ethical and religious concerns. The percentage of pregnancies is higher than after in-vitro fertilization. When efficacious, this approach allows additional spontaneous conceptions without renewed therapy and the course of pregnancy and the type of delivery will not differ from those in a normal population.

Challenge tests in the diagnosis of latex allergy.

The aim of this study is to evaluate the sensitivity, specificity and safety of challenge tests and their usefulness in the diagnosis of latex allergy. Forty adult subjects (F/M = 34/6, aged 18-66 yrs) with a history of adverse reactions after latex exposure and positive prick test and/or specific IgE to latex were enrolled. They were compared with 20 control subjects. They underwent provocative (cutaneous, mucous-oral, sublingual, conjunctival, nasal, bronchial, vaginal) tests. Symptoms and drug scores were recorded for each patient during challenges. All patients reacted to at least one of the following: cutaneous, nasal and conjunctival tests. No systemic reactions requiring epinephrine occurred. Of the challenges, the vaginal test resulted as the safest, but it had low sensitivity and many limits related to the procedure. According to our data, bronchial and nasal tests had the highest sensitivity (76% and 82% respectively), and were more precise than other tests in determining latex exposure and symptoms, but the bronchial test also presented the highest rate of risk. Mucous and cutaneous tests resulted as the most reliable. For all the tests, specificity and positive predictive value were 100%. All control subjects resulted negative to all challenges. There were no statistically significant changes in skin and serologic tests between the first and second visits. Correlations between MIS and skin tests and between MIS and serum tests were not found. Challenges can be considered safe diagnostic procedures. Tests that most faithfully reproduce natural exposure, on the basis of a patient's history, are preferable.

Isolated and preclinical impairment of left ventricular filling in insulin-dependent and non-insulin-dependent diabetic patients.

BACKGROUND: Diabetes mellitus can induce a pattern of myocardial pathology known as specific diabetic cardiomyopathy, even if this is not clearly specified. HYPOTHESIS: The aim of our study was to evaluate the presence of preclinical myocardial damage in insulin- and non-insulin-dependent diabetic patients and controls by assessment with Doppler echocardiography. METHODS: Twenty insulin-dependent diabetic (IDDM) patients, 10 non-insulin-dependent diabetic (NIDDM) patients, and 12 healthy individuals (C) as controls, matched for age, gender, and without overt cardiovascular disease, were assessed in this study. RESULTS: Systolic function parameters presented normal values in the three groups, with the exception of a slight reduction in ventricular volume indices in the NIDDM group. Diastolic function was clearly impaired in both groups of patients versus that in healthy controls. In particular, ventricular filling was impaired in the NIDDM compared with the IDDM patients, especially the peak early filling rate E (p < 0.001). Moreover, in the IDDM group, the duration of diabetes (p < 0.01) and glycosilated hemoglobin value (HbA1C, p < 0.02) were higher than in the NIDDM group. Multiple regression analysis showed a significant inverse correlation between HbA1C and peak late filling rate A (R2 = 0.28) in both groups of patients and a direct correlation between velocity time integral E and age, duration of diabetes, and HbA1C (R2 = 0.46). The two groups presented a small, homogeneous number of cases with initial microangiopathy and borderline autonomic neuropathy, associated with microalbuminuria. Doppler echocardiography showed an early impairment of left ventricular filling, as well as an early preclinical alteration of myocardial function in diabetic patients, especially in the NIDDM group. CONCLUSION: These early signs of cardiomyopathy could constitute a predisposing condition toward the high cardiac morbidity and mortality rate in diabetic patients.

Left ventricular function in insulin-dependent and in non-insulin-dependent diabetic patients: radionuclide assessment.

The aim of this study was to compare, by gated radionuclide angiography, systolic and diastolic ventricular function in insulin-dependent (IDDM) and non-insulin-dependent (NIDDM) diabetic patients without overt cardiovascular disease. The study population consisted of 20 IDDM patients (15 male, 5 female; 40.7 +/- 10.3 years), 14 NIDDM patients (9 male, 5 female; 47.0 +/- 7.5 years) and 12 healthy subjects (7 male, 5 female; 41.5 +/- 6.3 years) as a control (C) group. The duration of diabetes (DD) and glycosylated hemoglobin (HbA1C) levels were significantly higher in the IDDM patients. The ventricular ejection fraction and peak ejection rate (PER) were assessed by gated radionuclide left ventriculography and were similar in three groups, while the peak filling rate (PFR) was lower in the NIDDM patients compared to the IDDM patients (p < 0.05) and controlled healthy subjects (p < 0.01, IDDM = 3.39 +/- 1.14; NIDDM = 2.65 +/- 0.83; C = 3.55 +/- 0.73), the time to PFR was significantly more prolonged in the NIDDM group than in the IDDM (p < 0.05) and C groups (p < 0.05, NIDDM = 162 +/- 26; IDDM = 140 +/- 28; C = 142 +/- 23). The PFR/PER ratio was near the normal value (approximately equal to 1) in the IDDM patients and controlled subjects, while in the NIDDM patients it was reduced (approximately equal to 0.84 +/- 0.18). Seven IDDM and 4 NIDDM patients had borderline signs of cardiovascular autonomic neuropathy, unrelated to DD, HbA1C and scintigraphic parameters. Left ventricular systolic performance was substantially normal and similar in both the IDDM and NIDDM patients. Ventricular diastolic filling was impaired in the NIDDM patients, as shown by the decrease in PFR and in particular in the PFR/PER ratio. Our radionuclide data suggest that the NIDDM patients had a prevalent abnormality of ventricular diastolic performance, with respect to the IDDM patients, although the latter patients had higher DD and HbA1C values.

[Urinary tract infections: risk factors and therapeutic trends]. / Infezioni urinarie: fattori di rischio ed orientamenti terapeutici.

Urinary tract infections (UTIs) are very common in medical practice. Women have a high prevalence of UTIs, approximately 50 times higher than men. A large proportion of this prevalence is probably caused by anatomic and physical factors Chemical analysis of urine composition, examination of the urinary sediment and the bacterial colony counts are of great value for diagnosis and therapy. The patients may be benefit from antibiotic doses. In addition to trimethoprimsulfamethoxazole (TMP/SMZ), amoxicillin and cephalosporins, the authors observed a new drug: fluoroquinolones. These drugs derived by nalidixic acid and included: ciprofloxacin, enoxacin, lomefloxacin, norfloxacin, pefloxacin and rufloxacin. They are sinergistic against most Gram positives and negatives including Pseudomonas aeruginosa and Proteus mirabilis. Fluoroquinolone is an antibacterial agent that is effective in treating urinary tract infections. It is usually administered orally and is well absorbed after oral ingestion. Quinolones are preferable to TMP/SMZ because of their greater antibacterial activity that occurred in about 82% of women. A dose of quinolones (400 mg daily for 3 days) has been particularly effective in the treatment of UTIs. The amoxicillin-clavulanic acid can be used for treatment even if increased antibiotic resistance. The efficacy, relative safety and low cost of quinolones predispose to utilize its like the first treatment choice.

[Mycotic vulvovaginitis]. / Vulvovaginiti micotiche.

Vulvovaginitis is the most common clinical manifestation of fungal infections causing human mycoses; the incidence occurs in 10% of women, during pregnancy the incidence achieves 30% of cases. Candida albicans has resulted to be the most commonly isolated agent in patients with fungemia. In fact, Candida appears to be the species recovered in as many as 90% of cases. They are mainly the sexual activity, hormonal contraception and several pathologies such as diabetes mellitus and thyroiditis responsible for the pathogenesis of infection. The first symptom of this infection is usually pruritus associated to leukorrhea, dyspareunia and vulvovaginal irritation. Antifungal therapy may be required in more severe cases of vulvovaginal candidiasis. Candida species can be identified on isolation culture media including agar and on direct examination. Diagnosis can also be made through san immunologic examination. However, the authors confirm that the risk factors together with a correct diagnosis of the Candida etiological agent in the different species (albicans, glabrata, tropicalis, krusei) should be accurately investigated in order to give the correct therapeutical approach.

[Treatment in the event of antibiotic prophylaxis failure in gynecologic surgery. A retrospective study of 20 cases]. / Sul trattamento in caso di fallimento dell'antibiotico-profilassi in chirurgia ginecologica. Esame retrospettivo di 20 casi.

A retrospective study was performed of the type of treatment used in 20 patients undergoing gynecological surgery in whom antibiotic prophylaxis with Mezlocillin (2 g i.v.) had failed. Patients were subdivided into three groups: A) Initial therapy with Mezlocillin (8 patients, 2 g/die i.m.) or Cefotetan (2 patients, 2 g/die i.m.) and subsequent addition of Gentamicin (8 patients, 240 mg/die i.m.) or Tobramycin (2 patients, 200 mg/die i.m.) and subsequently Metronidazole (7 patients, 1.5 g/die per os). B) Therapy with Imipenem/Cilastatin (6 patients, 1.5 g/die i.m.). C) Therapy with Imipenem/Cilastatin (4 patients, 1.5 g/die i.m.) after a variety of antibiotics: Cotrimoxazole (Trimethoprim 160 mg/die and sulphamethoxazole 800 mg/die per os), Pefloxacin (800 mg/die per os), Cefotetan (2 g/die i.m.) and Mezlocillin (2 g/die i.m.). Time taken to lower temperature was shorter in Group B (3.5 days) compared to Group A (6.8 days) and Group C (10 days). Postoperative hospital stay was also shorter in Group B (9 days) compared to Group C (16.5 days) and Group A (11.1 days). The immediate administration of an antibiotic active against Gram+ and Gram- germs, aerobes and anaerobes is therefore useful in the event of failure of antibiotic prophylaxis, rather than the use in succession of associations of antibiotics with a limited spectrum.

Characterization of amino acid transport in human endothelial cells.

The transport of amino acids has been studied in human umbilical vein endothelial cells. Neutral amino acids enter human umbilical vein endothelial cells through three distinct agencies endowed with the characteristics of systems A, ASC, and L. Each system has been studied by evaluating the influx of preferential substrates. The influx of L-proline and 2-methylaminoisobutyric acid occurs through an Na(+)-dependent adaptively regulated trans-inhibited agency identifiable with system A. L-Threonine influx occurs mainly through a distinct Na(+)-dependent trans-stimulated pathway corresponding to system ASC. System L accounts for Na(+)-independent influx of L-leucine. These systems cooperate for the transport of L-glutamine, which is due mainly to system ASC, whereas the component due to the operation of system A increases upon amino acid starvation. No clear evidence was found for a glutamine-specific system ("system N"). Two systems, one Na+ dependent (system XAG-) and the other Na+ independent (system xc-), transport anionic amino acids. L-Arginine influx exhibits a poor dependence on extracellular Na+, whereas it is sensitive to conditions known to change membrane potential and to trans-stimulation by intracellular amino acids. These features are consistent with a process mediated by system y+ and may be of significance for the regulation of the intracellular concentration of L-arginine.