An observational retrospective evaluation of 79 young men with long-term adverse effects after use of finasteride against androgenetic alopecia.

Concern regarding adverse effects of finasteride is increasing. We aimed to determine the type and frequency of symptoms in men having long-term sexual and non-sexual side effects after finasteride treatment (a condition recently called post-finasteride syndrome, PFS) against androgenetic alopecia (AGA). Subjects were recruited at the Urology Unit of the Trieste University-Hospital, and from a dedicated website. Out of 79 participants, 34% were white Italians, mean age was 33.4 ± 7.60 years, mean duration of finasteride use was 27.3 ± 33.21 months; mean time from finasteride discontinuation was 44.1 ± 34.20 months. Symptoms were investigated by an ad hoc 100 questions' questionnaire, and by validated Arizona Sexual Experience Scale (ASEX) and Aging Male Symptom Scale (AMS) questionnaires. By ASEX questionnaire, 40.5% of participants declared getting and keeping erection very difficult, and 3.8% never achieved; reaching orgasm was declared very difficult by 16.5%, and never achieved by 2.5%. By the ad hoc questionnaire, the most frequent sexual symptoms referred were loss of penis sensitivity (87.3%), decreased ejaculatory force (82.3%), and low penile temperature (78.5%). The most frequent non-sexual symptoms were reduced feeling of life pleasure or emotions (anhedonia) (75.9%); lack of mental concentration (72.2%), and loss of muscle tone/mass (51.9%). We contributed to inform about symptoms of PFS patients; unexpectedly loss of penis sensitivity was more frequent than severe erectile dysfunction and loss of muscle tone/mass was affecting half of the subjects. Further studies are necessary to investigate the pathophysiological and biochemical pathways leading to the post-finasteride syndrome.

Accuracy of a portable glucose meter and of a Continuous Glucose Monitoring device used at home by patients with type 1 diabetes.

BACKGROUND: Patients with diabetes are recommended to self-monitor their blood glucose levels also at home. Accuracy of a hand-held glucometer and a Continuous Glucose Monitoring (CGM) device were comparatively evaluated. METHODS: Venous blood samples (for reference laboratory determinations; n=428) were collected from 18 type 1 patients (35-65 years old), immediately followed by capillary measurement (Bayer ContourLink meter) and CGM readings (Medtronic Paradigm). RESULTS: Laboratory values did not differ statistically from ContourLink and CGM readings, mean difference (±SD) being -0.05±1.06 mmol/L and 0.10±1.84 mmol/L glucose, respectively. A bias ((value-reference)/reference×100) ≥15% was observed in 27.7% and 54.9% of cases, respectively. Notably, below 3.9 mmol/L glucose (hypoglycemic threshold), an absolute error>0.8 mmol/L was found in 78.9% and 94.1% of cases. The absolute errors of the CGM device were inversely related to the rate of glucose change (r=0.598, p<0.001). CONCLUSIONS: A very large error was observed at the extreme glycemic values, which may lead to erroneous therapy. Consequently, performance of future portable glucometers should be focused in particular under hypo- and hyper-glycemia. Moreover, integrated CGM devices should not disregard the effect of the rate of blood glucose change on the sensor readings.

Soluble transferrin receptor values in top level soccer players and skiers: comparison with sedentary people and pitfalls of laboratory methods.

AIM: Hematological assessment is crucial in athletes: the risk of sports' anemia should be monitored with hematological parameters and iron metabolism tests. The aim of this study was to evaluate soluble transferrin receptor (sTfR) efficacy, as it is highly sensitive and specific and usually utilized in sport medicine for monitoring iron metabolism. METHODS: sTfR was studied using two immunological methods (IDeA Orion, and Biokit) on a group of professional athletes, together with hematological and iron metabolism parameters. Values have been compared with those of sedentary people, before and during competitive season. Athletes were 76 professional male soccer players plus 20 males and 14 females of the alpine ski Italian National Teams. RESULTS: The sTfR values in athletes are similar to those found in sedentary people. Different results have been observed between the two different methods: a bias of 0.37 mg/L was found comparing them. A significant correlation between sTfR and iron, transferrin saturation, and reticulocytes was found in skiers; there was no correlation with hemoglobin, erythrocytes, ferritin. In soccer players significant differences have been retrieved among different teams' distribution of data. CONCLUSIONS: The principal limit for using sTfR in sports medicine, but also in the general population, is the lack of standardization among methods. The quantitative differences in athletes between the two methods are high, although the behavior of the parameter is similar from the quality point of view. The differences between measured concentrations could influence the thresholds used in antidoping context.

High concentrations of inhibin A and inhibin B in ovarian serous cystadenoma: relationship with oestradiol and nitric oxide metabolites.

Inhibin production has been demonstrated in malignant epithelial ovarian tumours, but secretion of inhibins by benign cystadenoma has not yet been reported. The present study evaluated the concentrations of inhibin A and inhibin B and the relationship with oestradiol and nitric oxide metabolites in fluid collected from benign ovarian serous cystadenomas (n = 15). In addition, follicular fluid samples (n = 14) from women with regular ovulatory cycles undergoing ovarian stimulation for IVF were studied as a reference group. High concentrations of inhibin A (median = 89.3 ng/ml) and inhibin B (median = 116.1 ng/ml) were found in the cystic fluid of ovarian serous cystadenomas. These inhibin concentrations were even higher than in follicular fluid of stimulated follicles (inhibins A and B = 41.2 and 46.8 ng/ml respectively; P: < 0.001), whereas oestradiol was approximately 18-fold lower in cystic fluid than in follicular fluid (median = 34 versus 622 pg/ml, P: < 0.001). In ovarian cysts, the concentrations of inhibin A and oestradiol were inversely correlated (r = -0.678, P: = 0.008). Cystic fluid samples containing the highest concentrations of NO(2)(-)/NO(3)(-) (45-60 micromol/l) had lower inhibin A and higher oestradiol concentrations than those samples containing lower concentrations (10-25 micromol/l) of NO(2)(-)/NO(3)(-). It is concluded that high amounts of dimeric inhibins are present in ovarian serous cystadenoma. The source of inhibins and the determinants of the inverse association of inhibin A with oestradiol and nitric oxide remain to be determined.

Impairment of the mucosal immune system: IgA and IgM cleavage detected in vaginal washings of a subgroup of patients with bacterial vaginosis.

The integrity of the immunoglobulins in vaginal washings of patients with bacterial vaginosis was examined to answer the question of the lack of immune response against Gardnerella vaginalis cytolysin. Clinically diagnosed patients (n=100) were recruited and their vaginal washings examined by Western blotting. Many showed IgA and IgM partially or extensively degraded. According to the degradation pattern, the patients were subdivided into 4 subsets, from intact (score 0) to completely degraded IgA (score +3). Statistical analysis of the data showed a correlation between IgA degradation and absence of immune response to G. vaginalis cytolysin. The extent of IgA degradation correlated also with the sialidase (but not with the prolidase) activity level. All women showed intact IgG and human serum albumin and no trypsin-like activity. Patients with bacterial vaginosis having high sialidase activity and extensive IgA degradation in their secretions could incur more dangerous infections and adverse pregnancy outcomes.

Immunoglobulin A response against Gardnerella vaginalis hemolysin and sialidase activity in bacterial vaginosis.

OBJECTIVE: The aim of this study was to investigate the correlation between the immunoglobulin A immune response to Gardnerella vaginalis hemolysin and sialidase activity in vaginal fluids from patients with bacterial vaginosis. STUDY DESIGN: Nonpregnant women who were examined at a gynecologic clinic, in an age range of 18 to 62 years, were enrolled. The study population comprised 131 healthy volunteers, 32 women with bacterial vaginosis that was positive for immunoglobulin A to Gardnerella vaginalis hemolysin, 40 women with bacterial vaginosis that was negative for immunoglobulin A to Gardnerella vaginalis hemolysin, and 19 women with Candida vaginitis. Bacterial vaginosis was diagnosed by clinical criteria and Gram stain. RESULTS: Sialidase activity was present in 75% (54/72) of patients with bacterial vaginosis. Women having bacterial vaginosis and lacking a specific immunoglobulin A response had a significantly higher level of sialidase activity than patients who had an immune response against Gardnerella vaginalis hemolysin. Sialidase activity was detected in 87% (35/40) of the former subgroup of patients with bacterial vaginosis and in 59% (19/32) of women of the latter subgroup. No sialidase activity was measured in patients with candidiasis. Specificity of the assay for healthy controls was 95% (124/131 women without sialidase activity). CONCLUSIONS: Sialidases produced by Prevotella bivia and other microorganisms present in the microflora of patients with bacterial vaginosis are very likely a virulence factor not only by destroying the mucins and enhancing adherence of bacteria but also by impairing a specific immunoglobulin A immune response against other virulence factors such as cytotoxin from Gardnerella vaginalis.

Specific immune response against Gardnerella vaginalis hemolysin in patients with bacterial vaginosis.

OBJECTIVE: Our goal was to study the mucosal host response in bacterial vaginosis by evaluating the presence of a specific immune response elicited against the Gardnerella vaginalis hemolysin in vaginal fluids of patients and by verifying its correlation with usual criteria adopted to diagnose bacterial vaginosis. STUDY DESIGN: A total of 123 white women attending the gynecologic care unit for urogenital complaints or for screening of uterine malignancies (Papanicolaou test) aged from 20 to 60 years, nonmenstruating, were enrolled. Bacterial vaginosis was diagnosed by clinical criteria and a Gram stain score > 6. RESULTS: We performed the determination of the antibody response in vaginal fluid against the hemolysin produced by G. vaginalis, a common agent present in bacterial vaginosis. The purified G. vaginalis toxin was a suitable antigen for detecting the presence of an immune response in the vaginal fluids of patients with bacterial vaginosis regardless of the strain of G. vaginalis present. A specific immunoglobulin A response was detected in 60% of women with overt bacterial vaginosis (Gram stain score > 6) and in 18.5% of women with intermediate vaginal flora (Gram stain score 4 to 6). The specificity of the test was 91%. CONCLUSIONS: We found a correlation between the specific local immune response to G. vaginalis toxin and bacterial vaginosis. The highly purified form of the toxin is able to discriminate disorders from the opportunistic colonization by G. vaginalis.

Sequence-specific DNA recognition by the thyroid transcription factor-1 homeodomain.

The molecular basis for the DNA binding specificity of the thyroid transcription factor 1 homeodomain (TTF-1HD) has been investigated. Methylation and ethylation interference experiments show that the TTF-1HD alone recapitulates the DNA binding properties of the entire protein. Studies carried out with mutant derivatives of TTF-1HD indicate a precise correspondence of some of its amino acid residues with specific bases in its binding site, allowing a crude orientation of the TTF-1HD within the protein-DNA complex. TTF-1HD shows an overall geometry of interaction with DNA similar to that previously observed for Antennapedia class HDs, even though the binding specificities of these two types of HDs are distinct. We demonstrate that the crucial difference between the binding sites of Antennapedia class and TTF-1 HDs is in the motifs 5'-TAAT-3', recognized by Antennapedia, and 5'-CAAG-3', preferentially bound by TTF-1. Furthermore, the binding of wild type and mutants TTF-1 HD to oligonucleotides containing either 5'-TAAT-3' or 5'-CAAG-3' indicate that only in the presence of the latter motif the Gln50 in TTF-1 HD is utilized for DNA recognition. Since the Gln at position 50 is an essential determinant for DNA binding specificity for several other HDs that bind to 5'-TAAT-3' containing sequences, we suggest that utilization by different HDs of key residues may depend on the sequence context and probably follows a precise hierarchy of contacts.

Ionic factors regulating the interaction of Gardnerella vaginalis hemolysin with red blood cells.

We have studied the hemolytic properties of an exotoxin released by Gardnerella vaginalis (Gvh). We found that hemolysis induced by Gvh is modulated by the composition of the isotonic buffer in which the red cells are suspended. In particular, low pH enhances its lytic activity, whereas low ionic strength and divalent cations diminish it. The inhibitory effects of reduced salt concentration and divalent cations occur despite normal binding of the toxin to the cells. This suggests that some post-binding step is impaired. The toxin is also able to damage cholesterol-containing lipid vesicles, and even on these model membranes it is more active at low pH. From this point of view, Gvh has some similarity to Clostridium perfringens theta-toxin, a membrane-damaging toxin belonging to the family of 'thiol-activated' cytolysins produced by Gram-positive bacteria.

Pore-forming and haemolytic properties of the Gardnerella vaginalis cytolysin.

The pleomorphic bacterium Gardnerella vaginalis releases in the culture broth a haemolytic exotoxin (Gvh) which is probably a virulence determinant of this unique bacterium, implicated in gynaecological and urological disorders. This 59 kDa cytolysin was purified to homogeneity in just one chromatographic step directly from the culture supernatant, a final specific activity up to 1.9 x 10(6) HU mg-1 being obtained. The toxin-induced lesion on human erythrocytes results from the formation of a pore whose radius is approximately 2.4 nm. The damage is inhibited by osmotic protectants and shows a sigmoidal dose-response profile suggesting an aggregation process of haemolysin molecules on the target membrane to create the functional lesion. The extent and the kinetics of haemolysis are strongly dependent on temperature and an activation energy of 64.0 kJ mol-1 has been derived. Lipid membranes can be very efficient inhibitors of Gvh-haemolysis, being able to bind the toxin quite avidly. The inhibitory effect requires the presence of cholesterol and it is stronger when cholesterol is mixed with negatively charged phospholipids rather than with zwitterionic phospholipids, suggesting that a negative surface potential increases the affinity of the toxin for the lipid bilayer. The functional properties of Gvh have been compared with those of Clostridium perfringens thetatoxin (PFO) and Escherichia coli haemolysin (HlyA), which are representative of widespread haemolysins produced by Gram-positive and Gram-negative bacteria, respectively. The toxin shares several features with the family of the so-called 'sulphydryl-activated' cytolysins produced by Gram-positive bacteria, although Gvh does not truly belong to this family, being deactivated by beta-mercaptoethanol and being antigenically distinct from them. We report here for the first time the detection in the vaginal fluid of infected women of a specific IgA response against the toxin.

ELISA method for quantitative measurement of IgA and IgG specific anti-gliadin antibodies.

In this study we describe the application of an enzyme-linked immunosorbent assay (ELISA) for the measurement of anti-alpha-gliadin antibodies (AGA) in absolute units (microgram protein/ml). Enriched samples of IgA and IgG AGA were obtained by means of protein A chromatography after immunoaffinity purification of pooled sera from untreated celiac patients. No cross-reactivity with other food antigens (beta-lactalbumin, soya proteins, ovalbumin) was detected. The quantitative evaluation of protein content in IgA and IgG AGA samples obtained by immunoaffinity chromatography, was performed by means of ELISA sandwich method using a reference curve obtained with pure standard human immunoglobulins. Scalar concentrations of purified IgA and IgG were then used to obtain a reference standard curve by means of an ELISA method. Such standard curve was utilized for titrating AGA in 214 sample sera. The minimal detectable concentration of IgA and IgG AGA was 0.02 micrograms/ml. The reproducibility of within- and between-assay resulted very good for IgA-AGA and acceptable for IgG-AGA. The method here described seems to be satisfactory not only for quantitative diagnostic purposes in routine screenings but also in epidemiological studies. Moreover, it can constitute a suitable way to solve practical problems of quality control of AGA ELISA assay.

NMR structural characterization of the reaction product between d(GpG) and the octahedral antitumor complex trans-RuCl2(DMSO)4.

The reaction between the antitumor octahedral complex trans-RuCl2(DMSO)4 and d(GpG) leads to the formation of a stable compound characterized by a covalent bifunctional coordination of the bases to the metal center. The structure of the compound has been fully characterized by NMR and molecular modeling studies, showing the presence of two N7-coordinated guanine moieties in a head to head conformation, two dimethyl sulfoxide molecules, and one halogen atom in the coordination sphere of the ruthenium. The glycosidic chi angles are essentially in the anti range, the sugar puckering of the 5'G is 3'-endo (100% N), whereas that of the 3'G is more flexible but mainly in 2'-endo conformation (85% S), the two bases are strongly destacked. The compound shows structural features which are surprisingly similar to those exhibited by the corresponding cisplatin complex, indicating that such a way of interaction with DNA is not exclusive to Pt or to metals with square planar coordination geometries.

Reaction of the octahedral antitumor complex trans-RuCl2 (DMSO)4 with 2'deoxyguanosine.

The reaction of the antitumor octahedral complex trans-RuCl2(DMSO)4 with 2'deoxyguanosine leads to the reversible formation of two diastereoisomeric monoadducts and one biadduct. This shows that it is possible to accommodate two purine bases in a cis configuration in an octahedral transition metal complex which exhibits antiblastic activity. All the product compounds are characterized by a guanine moiety coordination via the N7 atom. A marked decrease (about two pK units) is observed for the N1H pKa of the coordinated guanine moieties. The reversibility of the monodentate binding could explain the low toxicity of the ruthenium(II) complexes.

A dot immunobinding assay to detect anti-alpha-gliadin antibodies in celiac disease.

A dot immunobinding assay to detect anti-alpha-gliadin-specific antibodies in the sera or whole blood of enteropathic patients is described here. The method is based on the adsorption of alpha-gliadin as a spot onto nitrocellulose sheets. After incubation with the patient sample, the detection of specific antibodies is performed with alkaline phosphatase-conjugated goat anti-human (IgA or IgG) antibodies. Twenty-one celiac serum samples together with 18 enteropathic or disease controls and 44 healthy controls were analyzed. The classical ELISA test and the dot test gave comparable results. The dot test gave reliable result even when whole blood was tested. The method proved to be simple and sensitive.

[Anti-alpha-gliadin antibodies. Sensitivity, specificity and correlation with blood xylose test in the 3 diagnostic stages of celiac disease in children]. / Anticorpi antialfagliadina. Sensibilità, specificità e correlazione con la xilosemia da carico nelle tre fasi diagnostiche della malattia celiaca nel bambino.

The specificity and sensibility of IgA and IgG alfagliadin antibody test (AaGA) for screening, diagnosis and follow-up of childhood coeliac disease has been evaluated. We have compared AaGA test to D-xylose blood test and at last we have examined the false positive and negative results given by the test. Two groups of subjects were considered: 1) 90 children with untreated coeliac disease (21 newly diagnosed (I stage), 50 in gluten withdrawal (II stage), 19 in challenge (III stage); 2) 255 disease controls including: 157 healthy controls; 31 children with gastroenterological disorders other than coeliac disease; 31 children with food allergy and atopic dermatitis; 36 children with "constitutional" short stature (without GH deficiency and with normal intestinal mucosa). The sensibility of AaGA test in the first stage of coeliac disease has been of 95.2% for the IgG class antibody and 90.4% for the IgA class; on the other hand the showed a specificity of 83.6% for IgG class antibody and 96.9% for IgA class. In only two newly diagnosed coeliac children we have found false negative results: in the first case the patient was IgA-deficient, in the second the age was above 3 years. AaGA IgA resulted positive only in the 12.9% of the group of gastroenterological and atopic controls; particularly most cases were affected by multiple food allergies and two patients by chronic autoimmune disease of small intestine.(ABSTRACT TRUNCATED AT 250 WORDS)

Serum IgA and IgG antibodies to alpha-gliadin: comparison between two ELISA methods.

We have developed two ELISA methods, i.e., enzyme immunoassay (EIA) and fluorescence immunoassay (FIA), for the semiquantitative detection of specific IgA and IgG antibodies directed against alpha-gliadin. The tests differ only for the enzyme substrate and, when optimized, could be used in large routine screening of celiac disease. Several serum samples from patients with celiac disease and gastrointestinal disorders as well as from control subjects were tested. Both methods gave good correlation with clinical data, were easily performed and had some specificity features, while FIA proved to be more sensitive.