Role of inflammation, oxidative stress, and autonomic nervous system activation during the development of right and left cardiac remodeling in experimental pulmonary arterial hypertension.

This study investigated the impact of experimental pulmonary arterial hypertension (PAH) progression by evaluating morphometric and functional parameters, oxidative stress, autonomic nervous system (ANS) activation, and inflammation in the right (RV) and left (LV) ventricles. Male rats were first divided into two groups: monocrotaline (MCT) and control. The MCT group received a single MCT injection (60 mg/kg, intraperitoneal), while control received saline. The MCT and control groups were further divided into four cohorts based on how long they were observed: 1, 2, 3, and 4 weeks. Animals were submitted to echocardiographic and hemodynamic analysis. RV and LV were used for morphometric, biochemical, and histological measurements. Autonomic modulation was evaluated by cardiac spectral analysis, considering two components: low frequency (LF) and high frequency (HF). Lung and liver weight was used for morphometric analysis. MCT induced 100% mortality at 4 weeks. In the RV, disease progression led to mild inflammation and enhanced reactive oxygen species (ROS) in week 1, followed by moderate inflammation, ROS production, and hypertrophy in week 2. By week 3, there was moderate inflammation, oxidative stress, and ANS imbalance, with development of right heart dysfunction. LV biochemical changes and inflammation were observed at week 3. The initial changes appeared to be related to inflammation and ROS, and the later ones to inflammation, oxidative stress, and ANS imbalance in MCT animals. This study reinforces the severity of the disease in the RV, the late effects in the LV, and the role of ANS imbalance in the development of heart dysfunction.

Effects of aerobic exercise training on metabolism of nitric oxide and endothelin-1 in lung parenchyma of rats with pulmonary arterial hypertension.

Pulmonary arterial hypertension (PAH) is characterized by vasoconstriction and proliferative obstruction of pulmonary vessels, which promotes a progressive increase in pulmonary vascular resistance (PVR). The effect of exercise training on oxidative stress, metabolism, and markers of nitric oxide (NO) and endothelin-1 (ET-1) was analyzed in the lung tissue of rats with PAH induced by monocrotaline (MCT).Twenty-four Wistar rats were divided into four groups (5-7 animals): sedentary control (SC), sedentary MCT (SM), trained control (TC), and trained MCT (TM). The TC and TM groups participated in a treadmill training protocol (60% VO2 max) for 5 weeks, 3 weeks of which were performed after the injection of MCT (60 mg/kg i.p.) or saline. MCT administration promoted an increase in PVR and right ventricle hypertrophy, and reduction of right ventricle systolic function assessed by echocardiography. These changes were not improved by exercise training. The activity of NO synthase was reduced in the animals of the TC, TM, and SM groups. No significant differences were found in total nitrite concentration and expression of endothelial NO synthase. Moreover, the TM group showed strong staining for iNOS and nitrotyrosine, suggesting an increase in oxidative stress in these animals. In parallel, reduced expression of type B ET-1 receptors was noticed in the SM and TM groups in comparison to controls. In conclusion, the aerobic training protocol was unable to mitigate changes in the metabolism of NO and ET-1, probably because of the disease severity in these animals, especially in the TM group.

First evidence of pep solar neutrinos by direct detection in Borexino.

We observed, for the first time, solar neutrinos in the 1.0-1.5 MeV energy range. We determined the rate of pep solar neutrino interactions in Borexino to be 3.1±0.6{stat}±0.3{syst} counts/(day·100 ton). Assuming the pep neutrino flux predicted by the standard solar model, we obtained a constraint on the CNO solar neutrino interaction rate of <7.9 counts/(day·100 ton) (95% C.L.). The absence of the solar neutrino signal is disfavored at 99.97% C.L., while the absence of the pep signal is disfavored at 98% C.L. The necessary sensitivity was achieved by adopting data analysis techniques for the rejection of cosmogenic {11}C, the dominant background in the 1-2 MeV region. Assuming the Mikheyev-Smirnov-Wolfenstein large mixing angle solution to solar neutrino oscillations, these values correspond to solar neutrino fluxes of (1.6±0.3)×10{8} cm{-2} s^{-1} and <7.7×10{8} cm{-2} s{-1} (95% C.L.), respectively, in agreement with both the high and low metallicity standard solar models. These results represent the first direct evidence of the pep neutrino signal and the strongest constraint of the CNO solar neutrino flux to date.

Exercise training decreases oxidative stress in skeletal muscle of rats with pulmonary arterial hypertension.

The effects of exercise training on oxidative stress in gastrocnemius of rats with pulmonary hypertension were studied. Four groups were established: sedentary control (SC), sedentary monocrotaline (SM), trained control (TC), trained monocrotaline (TM). Exercise was applied for 4 weeks, 5 days/week, 50-60 min/session, at 60% of VO2 max. Right ventricular (RV) pressures were measured, heart and gastrocnemius were removed for morphometric/biochemical analysis. Lipid peroxidation (LPO), H2O2, GSH/GSSG, and activity/expression of antioxidant enzymes were evaluated. Increased RV hypertrophy, systolic and end-diastolic pressures (RVEDP) were observed in SM animals, and the RVEDP was decreased in TM vs. SM. H2O2, SOD-1, and LPO were higher in the SM group than in SC. In TM, H2O2 was further increased when compared to SM, with a rise in antioxidant defences and a decrease in LPO. GSH/GSSG was higher only in the TC group. Exercise induced an efficient antioxidant adaptation, preventing oxidative damage to lipids.

Precision measurement of the (7)Be solar neutrino interaction rate in Borexino.

The rate of neutrino-electron elastic scattering interactions from 862 keV (7)Be solar neutrinos in Borexino is determined to be 46.0±1.5(stat)(-1.6)(+1.5)(syst) counts/(day·100 ton). This corresponds to a ν(e)-equivalent (7)Be solar neutrino flux of (3.10±0.15)×10(9) cm(-2) s(-1) and, under the assumption of ν(e) transition to other active neutrino flavours, yields an electron neutrino survival probability of 0.51±0.07 at 862 keV. The no flavor change hypothesis is ruled out at 5.0 σ. A global solar neutrino analysis with free fluxes determines Φ(pp)=6.06(-0.06)(+0.02)×10(10) cm(-2) s(-1) and Φ(CNO)<1.3×10(9) cm(-2) s(-1) (95% C.L.). These results significantly improve the precision with which the Mikheyev-Smirnov-Wolfenstein large mixing angle neutrino oscillation model is experimentally tested at low energy.

[Effectiveness of the treatment with beta-glucan in the HPV-CIN 1 lesions]. / Efficacia del carbossimetilbetaglucano nella regressione delle alterazioni citologiche cervicali di basso grado HPV correlate.

AIM: The aim of the study was to evaluate the effectiveness of the beta-glucan in women with abnormal cytology, including the women with a positive screening for ASCUS-LSIL furtherly divided in women with positive cytology (ASCUS or LSIL) and negative colposcopy and women with abnormal cytology, positive colposcopy and human papilloma virus (HPV)-CIN1 hystology who opted for follow-up. METHODS: From September 2007 to December 2008, 60 women with ASCUS-LSIL diagnosis were recruited at the ambulatory of Lasersurgery and Cervico-Vaginal Patology, Department of Gynecology and Obstetrics of Policlinico Umberto I of Rome. The women was subdivided in two groups: 1) women with cytological diagnosis of ASCUS or LSIL and negative colposcopy; 2) women with abnormal cytology, positive colposcopy and HPV-CIN1 histology, who opted for follow-up. All the women were treated with two cycles of a daily topical application of beta-glucan for 20 consecutive days with a suspension of 10 days. The effects of beta-glucan were analyzed with colposcopy and cytology at 3.6 and 12 months from the beginning of the therapy. RESULTS: After 3 months of treatment, of the 30 women with positive cytology and negative colposcopy, 80% with ASCUS diagnosis resulted negative, 35% with LSIL diagnosis resulted negative; after 6 months 100% with ASCUS diagnosis resulted negative, 70% with LSIL diagnosis resulted negative; after 12 months 85% with LSIL diagnosis resulted negative. Of the 30 women with positive cytology, positive colposcopy and HPV-CIN1 histology after 3 months 20% resulted negative, after 6 months 60% resulted negative and after 12 months 80% resulted negative. The persistence of the HPV-CIN1 histology was verified in the 13% of the women. For these women the definitive treatment was the TFD. CONCLUSION: Our study demonstrate the effectiveness of the treatment with beta-glucan in the women with ASCUS-LSIL lesions and HPV-CIN1 lesions, increasing of the regressions rate after 12 months of the treatment of the 15-20%.

Scalable Super-Resolution Synthesis of Core-Vest Composites Assisted by Surface Plasmons.

The behavior of composite nanostructures depends on both size and elemental composition. Accordingly, concurrent control of size, shape, and composition of nanoparticles is key to tuning their functionality. In typical core-shell nanoparticles, the high degree of symmetry during shell formation results in fully encapsulated cores with severed access to the surroundings. We commingle light parameters (wavelength, intensity, and pulse duration) with the physical properties of nanoparticles (size, shape, and composition) to form hitherto unrealized core-vest composite nanostructures (CVNs). Unlike typical core-shells, the plasmonic core of the resulting CVNs selectively maintains physical access to its surrounding. Tunable variations in local temperature profiles ≳50 °C are plasmonically induced over starburst-shaped nanoparticles as small as 50-100 nm. These temperature variations result in CVNs where the shell coverage mirrors the temperature variations. The precision thus offered individually tailors access pathways of the core and the shell.

Profile of pterostilbene-induced redox homeostasis modulation in cardiac myoblasts and heart tissue.

This study was designed to investigate the effect of pterostilbene (PTS) on cardiac oxidative stress in vitro, as this is a simple and promising methodology to study cardiac disease. Cardiac myoblasts (H9c2 cells) and homogenised cardiac tissue were incubated with the PTS and cyclodextrin (PTS + HPßCD) complex for 1 and 24 h, respectively, at concentrations of 50µM for the cells and 25 and 50µM for cardiac tissue. The PTS + HPßCD complex was used to increase the solubility of PTS in water. After the pretreatment period, cardiomyoblasts were challenged with hydrogen peroxide (6.67µM) for 10 min, while cardiac tissue was submitted to a hydroxyl radical generator system (30 min). Cellular viability, oxidative stress biomarkers (e.g. total reactive oxygen species (ROS), carbonyl assay and lipoperoxidation) and the antioxidant response (e.g. sulfhydryl and the antioxidant enzyme activities of superoxide dismutase, catalase and glutathione peroxidase) were evaluated. In cardiomyoblasts, the PTS + HPßCD complex (50µM) increased cellular viability. Moreover, the PTS + HPßCD complex also significantly increased sulfhydryl levels in the cells submitted to an oxidative challenge. In cardiac tissue, lipid peroxidation, carbonyls and ROS levels were significantly increased in the groups submitted to oxidative damage, while the PTS + HPßCD complex significantly reduced ROS levels in these groups. In addition, the PTS + HPßCD complex also provoked increased catalase activity in both experimental protocols. These data suggest that the PTS + HPßCD complex may play a cardioprotective role through a reduction of ROS levels associated with an improved antioxidant response.

A silicon carbide array for electrocorticography and peripheral nerve recording.

OBJECTIVE: Current neural probes have a limited device lifetime of a few years. Their common failure mode is the degradation of insulating films and/or the delamination of the conductor-insulator interfaces. We sought to develop a technology that does not suffer from such limitations and would be suitable for chronic applications with very long device lifetimes. APPROACH: We developed a fabrication method that integrates polycrystalline conductive silicon carbide with insulating silicon carbide. The technology employs amorphous silicon carbide as the insulator and conductive silicon carbide at the recording sites, resulting in a seamless transition between doped and amorphous regions of the same material, eliminating heterogeneous interfaces prone to delamination. Silicon carbide has outstanding chemical stability, is biocompatible, is an excellent molecular barrier and is compatible with standard microfabrication processes. MAIN RESULTS: We have fabricated silicon carbide electrode arrays using our novel fabrication method. We conducted in vivo experiments in which electrocorticography recordings from the primary visual cortex of a rat were obtained and were of similar quality to those of polymer based electrocorticography arrays. The silicon carbide electrode arrays were also used as a cuff electrode wrapped around the sciatic nerve of a rat to record the nerve response to electrical stimulation. Finally, we demonstrated the outstanding long term stability of our insulating silicon carbide films through accelerated aging tests. SIGNIFICANCE: Clinical translation in neural engineering has been slowed in part due to the poor long term performance of current probes. Silicon carbide devices are a promising technology that may accelerate this transition by enabling truly chronic applications.

Soy isoflavones, lactobacilli, Magnolia bark extract, vitamin D3 and calcium. Controlled clinical study in menopause.

AIM: Affective and behavioural disorders possibly concomitant to the vasomotor menopausal symptoms worsen quality of life. A rational formulation containing soy isoflavones (60 mg), lactobacilli (500 millions spores), calcium (141 mg) and vitamin D3 (5 microg) was added of Magnolia bark extract (60 mg) and magnesium (50 mg) (Estromineral serena, ES). The Magnolia extract active principles interact with GABA system and exhibit a sedative central action. Magnesium intervenes in enzymatic reactions of the energetic metabolism and protects the bone integrity. Aim of this controlled study was to compare the clinical activity and safety of ES versus calcium+vitamin D3 (Ca+D) in menopause. METHODS: A controlled, randomised, multicentre study was carried out in symptomatic menopausal women with sleep or mood alterations. Women received 1 tablet/day of ES or Ca+D for 24 weeks. Symptoms during the treatment and final judgements on efficacy and acceptability were evaluated. RESULTS: Eighty-nine women (44 ES and 45 Ca+D, mean age 53.8 years, in menopause since 56.6 months) participated to the study. Flushing, nocturnal sweating, palpitations, insomnia, asthenia, anxiety, mood depression, irritability, vaginal dryness, dyspareunia, and libido loss, significantly decreased in severity and frequency during ES versus Ca+D treatment even since the fourth week. Woman wellbeing (good/very good 66.7% vs 20%) judgement on efficacy (72.7% vs 17.1%) and acceptability (93.9% vs 31.4%) were significantly better for ES. CONCLUSIONS: The controlled study showed the efficacy of Magnolia extract and magnesium on psycho-affective and sleep disturbances in menopause, in addition to the effects of isoflavones on vasomotor symptoms. A global natural approach to menopause with ES evidenced its therapeutic usefulness and safety.

Studies on the nature of in vitro and in vivo photosensitization reactions by psoralens and porphyrins.

This study was directed to examine the role of type II (photodynamic) reactions involving the production of reactive oxygen species (singlet oxygen, superoxide anion, and hydroxy radicals) in in vitro and in vivo photosensitization reactions induced by skin photosensitizing chemicals. Several porphyrins and psoralens, as model compounds representing examples of endogenous and exogenous photosensitizers, were examined for their ability to (a) produce singlet oxygen and superoxide anions, (b) induce damage to membranes and associated microsomal P-450, (c) promote lipid peroxidation of microsomal lipids of liver and epidermal cells, and (d) induce skin photosensitization reactions in vivo. Dose-response study in vitro of singlet oxygen production in H2O and D2O and inhibition studies involving the production of singlet oxygen and superoxide anion by specific quenchers indicated significant production of singlet oxygen by porphyrins, about 5-20 times higher than psoralen at 10(-5) M and 10(-6) M concentration and irradiation dose of 1-5 J/cm2 of UVA (greater than 320-400 nm radiation). The comparative studies on aerobic photodegradation of microsomal P-450 of guinea pig epidermis and liver indicated a significantly greater destruction of P-450 with porphyrins than with psoralens. A membrane-lipid peroxidation study involving malondialdehyde production, using liver and epidermal microsomal fractions with and without porphyrins, psoralens, and UVA radiation, indicated 10-20 times increased production of malondialdehyde with UVA and porphyrins than with psoralens.

Characterization of superoxide dismutase from mammalian skin epidermis.

Superoxide dismutase provides a protective defense mechanism in cellular compartments against the potential cytotoxicity of superoxide anion generated by ultraviolet radiation. Little information is available about the nature of superoxide dismutase in mammalian skin. We report the isolation and characterization of superoxide dismutase from human, guinea pig, and mouse epidermis. Copper-zinc superoxide dismutase was detected in all the mammalian skin specimens examined. Manganese superoxide dismutase was detected in human and guinea pig epidermis but not in the newborn or adult albino CD1 mouse epidermis. Electrophoresis studies of the extracted and partially purified skin superoxide dismutase on polyacrylamide gel slabs in the presence of sodium dodecylsulfate showed the characteristic molecular weights for subunits of 16,500 for copper-zinc superoxide dismutase, and 23,500 for mangano superoxide dismutase. Studies under nondenaturing conditions revealed significant differences in the mobility of the enzymes, depending on the sources of superoxide dismutase. The mouse epidermal copper-zinc superoxide dismutase was found similar to the bovine liver copper-zinc superoxide dismutase used as an internal standard. The copper-zinc superoxide dismutase of human skin and guinea pig skin showed activity-stained bands characterized by a higher mobility than the same enzyme from mouse or bovine liver. Quantitative data using the beta-NADH oxidation method indicated a 5-10-fold lower content of superoxide dismutase in mammalian epidermis in comparison with other tissues examined during this study, or compared with reported values in the literature.

Plasma interleukin-1 beta and tumor necrosis factor concentrations in obsessive-compulsive disorders.

Plasma interleukin-1 beta (Il-1 beta) and tumor necrosis factor-alpha (TNF-alpha) concentrations were measured twice, at a 48-hour interval, in 27 drug-free obsessive-compulsive patients (12 women and 15 men) and in 27 sex-age-matched healthy controls. Il-1 beta and TNF-alpha concentrations were significantly lower in patients than in controls, whereas there were no differences in either group between men and women, between the samples of the two days, or, in the patients, between those who had and those who had not been previously treated with psychopharmacologic drugs.

DNA ploidy and HPV subtypes in cervical smears of HIV-sero-positive and negative patients.

Epidemiological studies have demonstrated that cervical HPV infection and precancerous lesions of the cervix are more common in HIV-seropositive patients. However little is known about the natural history of these lesions in this population. In the present study cervical smears from 36 patients, 18 HIV-seropositive women and 18 matched controls were evaluated with the aim of quantifying morphological alterations and to evaluate DNA ploidy and HPV subtypes. Cervical lesions in HIV-seropositive patients were diploid in 50% of the cases compared to 25% in controls. The only HPVs identified by ISH were types 16/18 and no significant differences were observed in the control population. In contrast, cytological evidence of HPV infection and dysplastic changes was greatly increased in smears from HIV patients compared to HIV-seronegative women. Less than 5% of the cells showed HPV associated changes in controls while 10% to 30% of the cells were affected in HIV-patients. We suggest that the Papanicolaou test should be effective for detecting cervical disease and for a close follow-up of this population. Moreover, while additional studies with larger population groups and different population bases are needed, these findings are suggestive of the possible use of morphological criteria for the identification of HIV-seropositive subjects.

Noradrenergic receptor sensitivity in obsessive-compulsive disorders: I. Growth hormone response to clonidine stimulation.

In 15 patients with obsessive-compulsive disorder (OCD) and in 15 healthy controls postsynaptic alpha-2-adrenoceptor sensitivity was examined by measuring the growth hormone (GH) response to growth hormone-releasing hormone (GHRH) and to clonidine stimulation. Basal values of GH and somatomedin-C (SMD-C) and mean GH responses to GHRH were the same in patients and controls, thus suggesting that a peripheral pathology of the somatotropic axis should not be present. GH responses to clonidine stimulation were blunted in patients suggesting that post-synaptic alpha-2-adrenoceptors are subsensitive, possibly due to higher than normal noradrenergic secretion.

Noradrenergic receptor sensitivity in obsessive compulsive disorder: II. Cortisol response to acute clonidine administration.

Sensitivity of pre-synaptic alpha-2- and post-synaptic alpha-1- and beta-adrenoceptors was investigated in 15 patients with obsessive-compulsive disorder (OCD) and in 15 age- and sex-matched healthy controls, by measuring cortisol responses to saline administration and to inhibition by clonidine (clon), a noradrenergic receptor agonist. Basal values of cortisol and responses to administration of saline and of clon were the same in patients and controls.

Polychlorinated biphenyls in sediments of the Venice Lagoon.

Twenty-four surficial sediment samples, representing different environments within the Venice Lagoon, were analyzed for polychlorinated biphenyls (PCBs) in order to assess degrees of pollution, sources, areal distribution, and potential risk for the environment. Concentrations in surficial sediments range from 2 to more than 2049 microg kg(-1) dw. The maximum is probably due to the discharge of industrial wastes into the Brentella Canal, within the first Industrial Area. Lagoon samples are 11-205 times less contaminated, being influenced by pollutants stored in the canals only occasionally. Congener profiles distinguish PCBs in two main categories. The baseline pollution is probably due to a variety of sources within the lagoon system, whereas less chlorinated PCBs were discharged into the Brentella Canal. Our estimates suggest that, due to high concentrations, toxic equivalents (TEQs) of PCBs are comparable to those relative to PCDD/Fs for the least contaminated samples, whereas they are lower in case of heavy PCDD/F contamination.

Accumulation of polychlorinated biphenyls in sediments of the Venice Lagoon and the industrial area of Porto Marghera.

Sediment cores, collected at seven sites in the Venice Lagoon and within the canals of the industrial area were analyzed for polychlorinated biphenyls (PCBs) in order to assess the chronology of pollution and its present trends. The surficial concentration of PCBs is very high (more than 2049 microg kg(-1)) only in the Brentella Canal, probably due to a very recent contaminating episode. Very high values downcore (up to 41,639 microg kg(-1)) can be found in different parts of the industrial area, especially in the canals Lusore-Brentelle and Salso. Lagoon samples are much less contaminated (2.7-123 microg kg(-1)), being influenced only occasionally by polluted sediments resuspended from the canals. Sediment chronology shows that the delivery of contaminants peaked in the 1970s-early 1980s, decreasing since at most sites. Congener profiles distinguish PCBs in two main categories: heavy congeners characterize a baseline pollution, probably due to a large variety of sources within the lagoon system, whereas a mixture of light PCBs was discharged into the canals Brentella and Salso.

Accumulation of polychlorinated dibenzo-p-dioxins and dibenzofurans in sediments of the Venice Lagoon and the industrial area of Porto Marghera.

Two sediment cores collected from the central Venice Lagoon and five from the canals of the nearby industrial area were studied in order to assess the temporal evolution of polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) delivery and present trends. The most polluted site is C12, in the Brentella Canal, with a total equivalent (TEQ) concentration of 2858 and 64,130 ng kg-1 at the surface and at 18-20 cm depth, respectively. High values can be also found in the Nord Industrial Canal and in the Salso Canal: 3000 and 2500 ng kg-1, respectively. Lagoon samples are much less contaminated, with TEQ downcore maxima of 22 and 53 ng kg-1, being influenced by pollutants stored in sediments of the canals only occasionally. Sediment chronology, based on 210Pb and 137Cs activity-depth profiles, shows that the highest fluxes occurred in the 1970s and early 1980s, and that pollution is now decreasing. Homologue profiles can distinguish PCDD/Fs from different sources: combustion for octachlorinated dibenzo-p-dioxin (OCDD) and stripping of VCM for octachlorinated dibenzofuran (OCDF) alone, whereas several activities carried out in the past within the industrial area can be responsible of the overall contamination by furans.

[Quantitative evaluation of endocervical cells in women with human papillomavirus infection]. / Valutazione quantitativa di elementi endocervicali in donne affette da Human Papilloma Virus.

UNLABELLED: Human papilloma virus (HPV) DNA in cells of endocervical origin has been demonstrated by in situ hybridization and Polimerase Chain Reaction (PCR). Similarly, the presence of HPV DNA has been shown in many cases of endocervical Adenocarcinoma. Morphological changes in squamous cells, specific for HPV infection, have already been described in previous reports, but similar specific alterations in the endocervical elements have not yet been reported. In this retrospective study we analyzed 30 endocervical smears from women with HPV infection, ascertained colposcopically, cytologically and/or histologically. Thirty endocervical samples have been taken from healthy women of the same age and hormonal status, as a control group. We evaluated maximum and minimum nuclear diameters, anisocariosis, chromatin pattern and cytoplasmic characteristics of at least 60 endocervical cells selected from each specimen. RESULTS: In the cases with HPV infection, the mean nuclear diameter of endocervical cells was increased and anisonucleosis was more evident. However, the most significant parameter was the chromatin distribution with a granular pattern in almost 90% of the cilindrical cells in more than 80% of the cases. The cytoplasm does not show any peculiar alteration of endocervical cells except for an unconspicous "alo" in some elements, but this pattern is not frequent. In conclusion, some of the parameters studied seem to have a diagnostic relevance in confirming HPV infection. This may suggest that this infection could be present even at endocervical site, and should be confirmed by molecular techniques or by microcolpohysteroscopy.