The EBV-DNA Can be Used as a Diagnostic and Follow-up Parameter of the Rhinopharyngeal Tumors in the Non-Endemic Population of the Western Sicily.

To evaluate whether EBV-DNA can be used as a diagnostic and follow-up parameter for nasopharyngeal tumors in a non-endemic population. The study was carried out in a university hospital. A retrospective study was conducted on 40 paraffin samples of histological preparations. EB-DNA was detected by real-time PCR technique. A prospective study was also conducted on a group of 30 patients who underwent nasopharyngeal biopsy for suspected nasopharyngeal carcinoma (NPC) by comparing EBV-DNA concentrations between the histological specimen and the serum. Quantification of genomic copies of EBV-DNA in serum and detection of anti-EBV antibodies was performed. In both groups the presence of high viral load of EBV-DNA was found in nonkeratinizing squamous cell carcinomas, in three cases of lymphepitomyoma and in 4 out of 6 cases of non-differentiated non-carcinoma lymph node metastases. squamous keratinizing cells. In all cases of NPC, an antibody pattern typical of reactivations (IgGVCA+, IgG-EA+, IgG-EBNA+) and IgA-EA-D, frequently positive in cases of NPC, has been highlighted. A good correlation between the high EBV-DNA charges and the histological diagnosis was highlighted. Our study also found that the assessment of viral EBV load can also be considered in the prognostic evaluation and in the follow-up of patients with NPC.

Impact of a new carrageenan-based vaginal microbicide in a female population with genital HPV-infection: first experimental results.

OBJECTIVE: The objective of this study was to assess safety, satisfaction, and anti-viral effect of a new carrageenan-based vaginal microbicide in a population of fertile female patients with genital human papillomavirus (HPV) infection. PATIENTS AND METHODS: Forty healthy and sexually active women aged 18-45 years with genital HPV infection were enrolled. Each subject was treated with a gel formulated with 0.02% carrageenan and Propionibacterium extract (CGP) (Carvir, Depofarma SpA, Mogliano Veneto, Treviso, Italy). The subjects were evaluated at baseline, after the I cycle of therapy and after the II cycle. At final status, treatment acceptability and satisfaction were evaluated using a 5-point Likert scale. Furthermore, the rate of HPV genital infection clearance at final follow-up was evaluated. These data were compared with the HPV genital infection clearance rate in a control group of patients not subjected to any therapy. RESULTS: Overall, 68 HPV infections were detected at baseline, among 40 subjects enrolled. The HPV 16 genotype was the most frequent (12%) followed by HPV 18 (10%), and HPV 53 (9%). At the end of the study, 22 (55%) patients were very satisfied, 14 (35%) were satisfied, 3 (7.5%) were uncertain, and only 1 (2.5%) was dissatisfied, with 0 very dissatisfied. Only 2 patients complained of a local adverse event. Analysing infection clearance at the end of the study, 60% of patients became HPV negative. Among these, 13 cases were high-risk HPV infection. There were 16 patients with persistent infection ("non-responders"). No patient developed a "de novo" genital lesion. After controlling for age, the intervention had an adjusted OR of 4.9 (95% CI 1.6-15.1) to clear HPV. CONCLUSIONS: The results of this work suggest that Carvir vulvovaginal microbicide gel is safe and well-tolerated. Furthermore, this experience supports the hypothesis that CG has a role in accelerating the normal clearance of genital HPV infection in women with a positive HPV-DNA test.

HPV infection in semen: results from a new molecular approach.

Human papillomavirus (HPV) is the agent of the most common sexually transmitted diseases causing a variety of clinical manifestations ranging from warts to cancer. Oncogenic HPV infection is the major cause of cervical cancer and less frequently of penile cancers. Its presence in semen is widely known, but the effects on fertility are still controversial. We developed a new approach to evaluate virus localisation in the different semen components. We analysed also the specific genotype localisation and viral DNA quantity by qPCR. Results show that HPV DNA can be identified in every fraction of semen: spermatozoa, somatic cells and seminal plasma. Different samples can contain the HPV DNA in different fractions and several HPV genotypes can be found in the same fraction. Additionally, different fractions may contain multiple HPV genotypes in different relative quantity. We analysed the wholeness of HPV DNA in sperm cells by qPCR. In one sample more than half of viral genomes were defective, suggesting a possible recombination event. The new method allows to easily distinguish different sperm infections and to observe the possible effects on semen. The data support the proposed role of HPV in decreased fertility and prompt new possible consequences of the infection in semen.

Analysis of persistence of human papillomavirus infection in men evaluated by sampling multiple genital sites.

OBJECTIVE: Although human papillomavirus (HPV) infection has been studied extensively in women, data on male infection are limited. The purpose of this study was to investigate persistence of HPV infection at multiple genital sites in men and to define potential associations with socio-behavioural characteristics. PATIENTS AND METHODS: Penile, urethral and seminal specimens were tested by the INNO-LiPA HPV system (Innogenetics) and a PCR assay. Persistence was defined as the detection of same HPV type at ≥ 2 consecutive visits. The Kaplan-Meier method and the log-rank test were applied to estimate the likelihood of persistence. RESULTS: A total of 50 men (median age: 33 years) were followed for a median of 14.7 months. Altogether, 49%, 36%, 26% and 11% of baseline HPV-positive men had 6-, 12-, 18- and 24-month persistent infection with any HPV type, respectively. The 6-, 12- and 18- month persistence was more common for oncogenic HPV infections; 24-month persistence was similar. The median duration of persistence was 21.7 months for any HPV. The median duration of persistence for any HPV type was significantly longer in the penile sample (22.5 months, 95% CI: 18.3-26.7) than the semen sample (15.3 months, 95% CI: 14.5-16.1). CONCLUSIONS: Over a third of type-specific HPV infections in men remained persistent over a 24-month period. The median duration of HPV infection was longer in penile samples compared to seminal samples. As being increasing the attention of HPV vaccination as a potential preventive approach also for men, it is imperative to obtain additional insight on natural history of HPV infection in men, particularly as far as incidence and duration are concerned.

Giant cell arteritis associated with chronic active Epstein-Barr virus infection.

Giant cell arteritis is an inflammatory vasculopathy that preferentially affects medium-sized and large arteries. A viral cause has been suspected but not confirmed in polymyalgia rheumatica and giant-cell arteritis. We report the case of a 81-year-old female who suffered from chronic active Epstein-Barr virus infection and developed giant cell temporal arteritis.

Zeolitized tuffs in pedotechnique for the reclamation of abandoned quarries. A case study in the Campania region (Italy).

The present study aims to assess the evolution of different proto-horizons as embryonic soils built by pedotechnologies for the reclamation and management of derelict and damaged lands, such as abandoned quarries. The model proto-horizons were assembled by utilizing coarse limestone gravel or zeolitized Phlegraean Yellow Tuff (PYT) as mineral components and commercial compost-amendments or a phosphorite-poultry manure mixture as organic matrices for growth of a pasture-grass under controlled conditions. The evolution of the model proto-horizons was followed by an evaluation of the stability and modification of the organic matter (OM) with reference to plant development. The results suggest that the natural carbonatic substrate occurring in limestone quarries was unable to sustain significant plant growth, while the PYT was suitable and efficient as a pedogenic substrate because it supported plant growth and induced a conspicuous accumulation of OM due to root activity. In particular, OM, including humic and non-humic components, greatly increased in the PYT treatment with the phosphorite-poultry manure mixture showing a concurrent trend toward humification. Conversely, an overall tendency toward degradation of OM was detected in the PYT model proto-horizon treated with commercial compost. Feasibility estimates show that quarry restoration costs appear reasonable where environmental impacts are high.

HPV infection in relation to OSCC histological grading and TNM stage. Evaluation by traditional statistics and fuzzy logic model.

We aimed to evaluate if in oral squamous cell carcinoma (OSCC) there is a relationship between histological grading (HG), TNM clinical stage and HPV infection; and to study the performance of fuzzy logic compared to traditional statistics, in the analysis of HPV status and correlates of OSCC. In cross-sectional analysis, the study group comprised 63 patients (mean age 68.89 years (SD +/-11.78), range (32-93); males 28 (44.4%), females 35 (55.6%)) with OSCC histologically diagnosed. HPV-DNA was studied in exfoliated oral epithelial cells by nested PCR (MY09/MY11 and GP5+/GP6+ primers). Data were analysed in parallel by traditional statistics with multivariate analysis and a fuzzy logic (FL) technique (membership functions as input, the ANFIS methodology, and the Sugeno's model of first order). HPV infection was detected in 24/63 (38.1%) of OSCC, as being HPV+ve 14/36 (38.9%) in G1, 7/18 (38.9%) in G2, and 3/9 (33.3%) in G3; HPV+ve 8/33 (24.2%) in Stage I, 9/12 (75.0%) in Stage II, 6/11(54.5%) in Stage III, and 1/7 (14.3%) in Stage IV. In both methods of analysis, no significantly increased risk of HPV infection was found for any HG score; whereas, TNM stage II was significantly associated to HPV infection (p=0.004; OR=9.375 (95% CI=2.030:43.30); OR'=11.148 (95% CI=1.951:43.30)), and, in particular, to primary tumour size T2 (p=0.0036; OR=7.812 (95% CI=1.914:31.890); OR'=9.414 (95% CI=1.846:48.013)); FL (% of prevision: 79.8; Root Mean-Square Error (RMSE): 0.29). No association was found between HPV infection and any demographical variable. Our findings show an association between HPV infection with TNM (stage II-T2), but not with histological grading of OSCC. Also, FL seems to be an additional effective tool in analysing the relationship of HPV infection with correlates of OSCC.

Proliferative verrucous vs conventional leukoplakia: no significantly increased risk of HPV infection.

Proliferative verrucous leukoplakia (PVL) is a very aggressive form of oral leukoplakia (OL) with high morbidity and mortality rates, hypothesised to be linked to HPV infection. This study aimed to determine the presence of HPV DNA in PVL in comparison with OL, and in relation to social-demographical variables (age, gender, smoking and drinking habits) in an Italian multi-centric hospital-based study. The study group consisted of 58 cases of PVL and 90 cases of OL as controls (47 homogeneous (H) and 43 non-homogeneous (non-H) form), both recruited from four Italian cohorts. HPV DNA was identified in exfoliated mucosal cells by nested PCR (nPCR) with MY09/MY11 and GP5+/GP6+ primer pairs and the HPV genotype determined by direct DNA sequencing. HPV DNA was found in 24.1% (14/58)of PVL and in 25.5% (23/90) of OL; there was thus no significant difference found between PVL and OL (both forms) for risk of HPV infection (OR=0.93; 95% IC:0.432-1.985). Similarly, in both groups of PVL and OL lesions, no statistic association was found between any demographical variable considered and HPV infection. HPV-18 was the most frequently detected genotype in all tissues, being found in 78.5% and 60.8% of HPV+ve PVL and OL, respectively. Other more rarely detected genotypes were HPV-16 (28.6% in PVL and 13% in OL), HPV-6 (17.4% in OL) and HPV-53 (8.8% in OL). PVL does not appear more likely to be associated to HPV infection than conventional OL lesions.

Detection of Mycobacterium tuberculosis complex in cattle by PCR using milk, lymph node aspirates, and nasal swabs.

The PCR technique was applied to the diagnosis of tuberculosis in live cattle, and both skin-test-negative and skin-test-positive animals were studied. DNA was taken from various sources including specimens of lymph node aspirates, milk, and nasal swabs. After slaughter and visual inspection, tissues such as lymph nodes, lungs, and udders from tuberculin reactors were tested by the same technique. Specific oligonucleotide primers internal to the IS6110 insertion element were used to amplify a 580-bp fragment. A 182-bp fragment was obtained by designating a nested PCR from the first amplification product. This fragment was cloned and sequenced, and after being labeled it was employed in dot blot hybridization. A total of 100 cattle were tested, and PCR analysis was performed using nasal swab, milk, and lymph node aspirate. Sixty skin-test-positive cows were also tested to detect mycobacterial DNA in tissue samples from lymph nodes, lungs, and udders, and the infection was confirmed in all of the animals. Using PCR analysis of tissue samples from slaughtered animals as a "gold standard" we calculated 100% values for sensitivity, specificity, and positive and negative predictive values for milk and lymph node aspirate samples. The respective values for nasal swab samples were 58, 100, 100, and 28%. The respective values for all of the samples were 74, 100, 100, and 35%, while for visual inspection the values were 81, 100, 100, and 58%, respectively. PCR analysis of specimens of lymph node aspirates, milk, and nasal swabs from skin-test-negative animals showed that 52% of these skin test results were false negatives. These animals, not being removed from the farms, represent a potential source of further infection.

Identification and characterization of a constitutive HSP75 in sea urchin embryos.

An antiserum against a hsp of the 70-kDa family was prepared, by means of a fusion protein, which was able to detect a constitutive 75-kDa hsc in the sea urchin P. lividus. This hsc was present both during oogenesis and at all developmental stages. A two-dimensional electrophoresis has revealed four isolectric forms of this 75-kDa hsc. The amino acid sequence of the fragment used to prepare the anti-hsp70 antibodies revealed a 43% identity with the corresponding part of sea urchin sperm receptor, and in mature eggs a brighter immunofluorescence was seen all around the cell cortex where the receptor for sea urchin sperm is localized. In oocytes the hsp75 was localized in the cytoplasms but not in the nuclei. In the embryos a higher hsp75 concentration was found in the portion facing the lumen of the cells which invaginate at gastrulation.

Liver targeting of adenine arabinoside monophosphate (ara-AMP) by coupling to lactosaminated human serum albumin.

Adenine arabinoside monophosphate (ara-AMP) is a potent antiviral agent against hepadnaviruses but its use in the treatment of chronic hepatitis B is hampered by severe neurotoxic side effects, which are dose dependent. In order to reduce these adverse reactions and to adopt the lysosomotropic approach to antiviral chemotherapy, ara-AMP was coupled to lactosaminated human serum albumin (L-HSA), a neoglycoprotein which specifically penetrates hepatocytes. In mice with Ectromelia virus hepatitis, ara-AMP coupled with L-HSA was selectively delivered to liver cells in which it was released in a pharmacologically active form. Moreover in woodchucks with WHV hepatitis and in patients with chronic HBV infection, coupled ara-AMP inhibited hepadnavirus replication at a dose (1.5 mg/kg/day) 3-6 times lower than the free drug. A clinical study using a 28-day period of treatment with conjugated ara-AMP at 1.5 mg/kg/day has now been started. In the first 6 patients the treatment has been completed. The conjugate inhibited virus growth without producing any side effects. L-HSA-ara-AMP conjugate must be given by intravenous infusion. New hepatotropic conjugates of ara-AMP have been recently prepared which could be administered by bolus intravenous injection or by intramuscular route. These complexes might assure a better compliance in patients with hepatitis B virus infection for a long lasting liver targeted antiviral treatment.

Quantitative analysis of wild-type and HBeAg minus hepatitis B viruses by a sequence-dependent primer extension assay.

The ratio between wild-type hepatitis B virus (HBV) and HBV mutant, unable to secrete "e" antigen (HBeAg minus HBV) appears to be an important determinant of the outcome of chronic hepatitis B. Quantitative analysis of wild-type and HBeAg minus HBVs in the blood could be useful to monitor chronic hepatitis B patients. We developed a solid-phase minisequencing assay for both viruses using a primer-guided incorporation of a single labeled nucleotide on an affinity captured biotinylated amplified HBV-DNA template. A standard curve was constructed by mixing increasing quantities of wild type and mutant virus DNAs. The detection of wild-type and HBeAg minus sequences, ranging from 10% to 90% of overall viremia, was linear and reproducible till 0.1 pg/microliter of serum HBV-DNA. The assay yields numerical values and the ratio of incorporated nucleotides defines the relative proportions (%) of the two viral sequences with accuracy. We tested the sensitivity and accuracy of the minisequencing on mixed end point dilutions of wild-type and HBeAg minus reference sera and amplified products. The feasibility and reproducibility of the assay were tested in 35 sera from 21 HBsAg positive patients with chronic hepatitis B using both minisequencing and oligo-hybridization assays. A high correlation was found between the two assays (r = 0.957 P < 0.0001). In conclusion, the minisequencing assay provides a precise and reproducible quantitative analysis of wild-type and HBeAg minus HBVs in clinical specimens. It is proposed to study the relations between HBV heterogeneity and the course of hepatitis B and its response to therapy.

Hepatitis B virus unable to secrete e antigen and response to interferon in chronic hepatitis B.

BACKGROUND: Anti-hepatitis e antigen-positive chronic hepatitis B is a progressive liver disease associated with precore mutant hepatitis B virus (HBV) and poor response to interferon. Therefore, precore mutant HBV may behave as an interferon-resistant virus. The relations between the prevalences of wild-type and precore mutant HBVs in baseline viremias and response to interferon were analyzed. METHODS: Sera from 115 patients (59 treated and 56 untreated, followed up for 30 months) were tested using a quantitative oligonucleotide hybridization assay. RESULTS: Spontaneous or interferon-induced recoveries were observed in 28.5% (6 of 21) and 47.3% (18 of 38) or in 0% (0 of 35) and 19% (4 of 21) of the patients with wild-type prevalent or mutant prevalent HBVs, respectively. Relapses occurred in 85.7% (12 of 14) and 19.4% (4 of 21) of treated patients with prevalent precore mutant and prevalent wild-type HBV, respectively (P = 0.0001). High precore mutant HBV levels (> 20% of total viremia) were associated with the lack of permanent response to interferon (P = 0.01). CONCLUSIONS: Precore mutant HBV can influence the response to interferon when it reaches significant serum levels (> 20% of total viremia). Therefore, chronic hepatitis B should be treated as early as possible in its natural history before precore mutant HBV is selected as a prevalent virus.

[Medical and social implications of abortion]. / Implicatiile medico-sociale ale avortului.

PIP: In the course of the evolution of human society the problem or idea of interrupting a pregnancy has been faced many times. Romania has adopted a mixed solution to the termination of pregnancy allowing abortions for medical, eugenic, and social reasons. The 1936 penal code allowed only medical abortion, but recent regulations have offered differing solutions. The old regulation not allowing termination of pregnancy or restricting it was in force with minor modifications until 1957. In 1966 a decree was issued that allowed women with 4 children an abortion for special reasons as determined by an abortion committee, but still therapeutic and strictly medical causes predominated. In 1985 a new regulation of medical law prohibited termination of normal pregnancy up to 28 weeks of gestation and infractions were punishable by law. Illegal induced abortion represents an antisocial manifestation that jeopardizes human relationships in society. Induced abortion occurs often in disintegrated family situations. The social implications of the phenomenon of birth are manyfold. Medical intervention is difficult because of the mutilating effect of abortion. The motives are a matter of reflection for physicians and jurists alike.^ieng

[Bicameral stimulation in a case of corrected transposition of the great vessels]. / Stimolazione bicamerale in un caso di trasposizione corretta dei grossi vasi.

A patient with Corrected Transposition of the Great Arteries (CTGA), mild incompetence of left A-V valve, complete atrioventricular block without associated anatomic lesions, in whom a bicameral permanent pace maker has been implanted, is described. The procedure of implantation did not present any particular problem; the stability of the electrocatheters was good. The exercise tolerance with DDD mode stimulation was normal and definitively better as compared to that achieved with ventricular stimulation at progressively higher stimulation frequency. The patient is now well and attending a normal physical activity.