High prevalence of latent tuberculosis infection in autoimmune disorders such as psoriasis and in chronic respiratory diseases, including lung cancer.

The early diagnosis and treatment of individuals harboring M. tuberculosis is key to ensuring the effectiveness of health programs aimed at the elimination of tuberculosis (TB). Monitoring for TB also has other important health care implications for the related immune pathology caused by the chronic inflammatory response to M. tuberculosis. Moreover, the recent introduction of biologic therapies for the treatment of several immune-mediated inflammatory diseases has shown unexpected high frequencies of reactivation of latent TB. The present cross-sectional study is aimed at estimating the prevalence of latent tuberculosis infection (LTBI) in different groups of subjects, either undergoing a routine program of screening for TB or a clinical monitoring of autoimmune or lung disorders, by analyzing their immune response in vitro to a pool of different M. tuberculosis antigens through an IFN-gamma-release assay (IGRA). We consecutively tested 1,644 subjects including health care workers (931), healthy immigrants from different countries (93), patients with a diagnosis of psoriasis (405), patients with lung inflammatory disease (60) or lung neoplasia (32) and a group of HIV-1 infected Italian subjects (120). The prevalence of IGRAs positive responses among health care workers was 8.9 percent. In comparison, significantly higher frequencies were found in healthy immigrant subjects (33.3%), similar to those found in inflammatory broncho-pneumopathies (34.5%) or lung cancer (29.6%). Interestingly, an unexpected high prevalence was also found in patients affected by psoriasis (18.0%), while HIV-infected subjects had values comparable to those of health care workers (10.8%). An age cut-off was determined and applied for each group by receiver operating characteristic (ROC) curves in order to perform the statistical analysis among age-comparable groups. Multivariate analysis showed that the age and clinical conditions such as having a diagnosis of psoriasis or a lung inflammatory disease were independent risk factors for developing an IGRA positive response. This study highlights an unprecedented high prevalence of IGRA positive responses among patients affected by psoriasis and emphasizes the need for a preliminary assessment of LTBI before the administration of any biologic therapy based on cytokine antagonists such as anti-TNF-alpha. Moreover, screening for LTBI should be routinely performed in the presence of a chronic pulmonary disease.

High risk human papillomavirus genotyping in clinical samples: evaluation of different commercial tests.

The aim of the present study is to compare the performance of several commercial human papillomavirus (HPV) tests in a cohort of 281 women. The hybrid capture II, the PreTect-HPV-Proofer, the linear array, and DR.HPVTMIVD were utilized to detect and type HPV in parallel with in-house PCR tests followed by direct automated sequencing or by sub-cloning and sequencing. The concordance levels along with other tests were evaluated with a Cohen's K value varying between 0.60 to 0.88, indicating good correlation with nearly perfect agreement between hybrid capture II, (HCII) and the linear array test. High sensitivity was recorded by the linear array and HCII with 100% (95% CI, 0.8021 to 1.0000) detection of cervical intraepithelial neoplasia (CIN) III by both methods. Conversely, the PreTect-HPV-Proofer showed high specificity with 12% (95% CI, 0.7966 to 0.9163) positivity on normal samples. The genotyping analysis showed that agreement among tests was only low to moderate with great differences between different HPV types. Multiple infections were detected with poor concordance and sub-cloning assays revealed the presence of a lower number of HPV in comparison to the other methods. In summary, the use of different HPV tests applied to the same group of cervical smears may possibly lead to incongruent results, suggesting the need to standardize type-specific sensitivity of genotyping methods and the need to evaluate their accuracy in detecting multiple HPV infections. This would be a prerequisite for the use of genotyping assays in cervical cancer screening programs.

Predictors of human papilloma virus (HPV) infection in Italian women.

HPV infection is a "necessary cause" of cervical cancer and it is sexually transmitted. Due to upcoming mass vaccination investigation on risk factors for infection is the basis to implement prophylactic strategy even in older women. The aim of the study was to evaluate predictors of high-risk (HR) HPV infection in adult women. Between 2006 and 2008, 100 women aged >18 years, with no previous treatment for cervical lesions, were screened for HR HPV infection in Rome, Italy. Risk factors for HPV infection were investigated through a questionnaire including: ethnicity, religion, education, marital status, sexual behavior, gynecological and obstetrical history, smoking and alcohol intake. Multivariate analysis identified the "never married-separated/divorced" status (OR: 3.38; 95% CI: 1.14-10.12) as predictor of HPV infection, while having a higher age at the first sexual intercourse (FSI) shows a protective effect (OR: 0.84; 95% CI: 0.71-1.00). A trend for the association between the infection and having more than three lifetime partners was also observed (OR: 2.57; 95% CI: 0.86-7.71). No significant association was found for other demographic characteristics investigated. These findings provide a contribution in the knowledge of an adult population defining a "high-risk" sexual behavioral profile and could be helpful to target prophylactic strategies in older woman.

HPV genotypes concordance between sex partners.

The HPV genotype concordance in the sexual couples could support the sexual viral transmission of HPV infection. The present study contains a case-report of a stable Italian sex couple harbouring the same five HPV genotypes in their genital samples. The female partner, affected by vulvar condilomatosis, evidenced positivity in her cervicovaginal scraping with high risk HPV DNA Hybrid Capture 2 test and was negative at liquid-based performed Pap Test and at colposcopic examination. The male partner was clinically healthy regarding his external genitalia. In both male and female genital scrapings, the following HPV genotypes were detected by means of a PCR-based assay: 6, 16, 53, 73 and 84. This considerably high genotype concordance does not appear to be casual and supports, in our opinion, the hypothesis that genital HPV types are sexually transmitted agents

Incidence of human papillomavirus infection in oral leukoplakia. Indications for a viral aetiology.

Epidemiological and experimental studies have indicated a possible role of human papillomavirus (HPV) in the etiopathogenesis of oral premalignant lesions and tumors. The aim of this study was to establish the incidence of HPV infection and the typing of genotypes in some patients with nonmalignant oral cavity pathology and healthy subjects. We selected 80 subjects affected by the following pathologies: oral leukoplakia (20), squamous cell papilloma (6), various forms of stomatitis (30), lichen planus (15), burning mouth syndrome (BMS, 9). Ten healthy subjects were used as control. The patients underwent a brushing directly on the lesion or on the lower gums and on the dorsal surface of the tongue and a new, sensitive method of in situ hybridization (ISH) with colorimetric signal amplification was used for HPV detecting. The samples were tested with a Mix probe, which identifies most of the HPV genotypes and, in positive cases, the specific probe for the 6-11, 31-33-51 and 16-18 genotypes, was used. Leukoplakia and papilloma were , therefore, associated with the HPV infection, differently from the other pathologies and healthy oral mucosa (chi-squared < 0.005). In conclusion, according to our findings, HPV is a specific risk factor for the development of oral premalignant lesions.

Clinically meaningful response to the EGFR tyrosine kinase inhibitor gefitinib ('Iressa', ZD1839) in non small cell lung cancer.

Preclinical research and ongoing clinical trials are validating epidermal growth factor receptor (EGFR) as a suitable molecular target in cancer therapy. Here we report the case of a 39-year-old non-smoking man with heavily pretreated stage IV bronchioloalveolar NSCLC who was treated with gefitinib ('Iressa', ZD1839) at the dose of 250 mg/day, orally. Symptomatic relief was already detectable after 2 weeks of gefitinib therapy, pulmonary function tests readily improved, performance status went from 2 to 0 within 8 weeks, CT scan performed after 10 weeks of treatment showed nearly complete resolution of the clinical picture, and clinical remission was maintained up to 32 weeks. This case-report illustrates the potential of EGFR tyrosine kinase inhibition to induce clinically meaningful responses in heavily pretreated, advanced NSCLC patients and suggests that participation of such patients into clinical trials of signal transduction inhibitors should be encouraged.

Colourimetric signal amplification of in situ hybridization assay for human papillomavirus DNA detection in cytological samples.

Signal amplification of In Situ Hybridization (ISH) can be obtained by Catalyzed Reporter Deposition (CARD), the catalyzed deposition of biotinylated tyramide at the location of the labelled probe. We analyzed 156 cervico/vaginal samples in order to evaluate whether ISH-CARD improves conventional ISH technique in detecting Human Papillomavirus (HPV) on cytological smears and can be used in clinical practice. 126 patients were clinically suspected to have a squamous intraepithelial lesions (SIL) and 30 were negative control-patients. We performed a HPV test screening on these 156 patients with both methods. The screening positive cases were tested for the presence of subtypes of HPV: 6/11, 31/33/51 and 16/18, using only the ISH-CARD. We found a significant increase of positivity with the amplified system, ISH: positive = 50 cases, ISH-CARD: positive = 120 cases, even in many cases cytologically negative. Many cases showed coinfections with different types of HPV. We observed overlapping results in some cell lines harbouring specific HPV subtypes (SiHa, CaSki and HeLa cells), whereas there was no reaction in the C33A cells, negative for HPV presence. This method enables the detection and the typing of HPV in cytological smears, represents a useful tool for clinical purposes and even identifies occult HPV infections.

Interlaboratory reproducibility of the immunocytochemical assessment of oestrogen and progesterone receptors and proliferative activity in fine needle aspiration of breast cancer.

The purpose of this study was to establish the interlaboratory reproducibility of immunocytochemical analysis of oestrogen (ER) and progesterone (PR) expression and Mib1 growth fraction on fine needle aspiration (FNA) smears. A set of 44 immunostained slides for ER, PR and Mib1 were randomly selected from the archives of the Center for the Study and Prevention of Cancer (CSPO) of Florence, Italy, and submitted for reading to 6 Italian laboratories. The generalized kappa statistic was used as an indicator of agreement among the six laboratories. A good correlation for ER and PR was evident. For Mib1 the results showed some discrepancies. In addition to adequate standardization of procedures, these data confirm that the reliability of the immunocytochemistry is strictly linked to accurate analysis of the results.

Restoring function with porcelain laminate veneers.

The ability of porcelain veneers to be conservative in dentistry has tremendous advantages for patients. Being less invasive with both the gingiva and pulp results in better gingival health, easier maintenance, and less postoperative sensitivity. The use of porcelain laminate veneers to reestablish function and enhance aesthetics results in minimally invasive restorations. The patient was very pleased with the results. With proper care, she will have many years of beauty and newly restored function.

Immunocytodiagnosis of atypical hyperplasia and endometrial carcinoma in post-menopausal women.

In the present study we have evaluated whether monoclonal antibodies (MAbs) B72.3 and AR-3 which display, on histological preparations, a differential reactivity with normal and transformed endometrium, could be a useful adjunct to endometrial cytology in the identification of pre-neoplastic and neoplastic conditions. Immunocytochemical (ICC) tests, using the 2 reagents, were performed on normal cycling endometrium and on hyperplastic and malignant lesions collected by the endocyte technique both from 86 surgically resected specimens and from 62 postmenopausal symptomatic and asymptomatic outpatients. The results obtained showed that the combination of the 2 MAbs can complement conventional morphology in the identification of pre-malignant atypical lesions and endometrial carcinoma of unclear cytological features, thus allowing a selection of those patients who are candidates for fractional curettage.

Expression of CAR-3 and TAG-72 macromolecules in normal and transformed endometrium: potential diagnostic application in postmenopausal patients.

Mass cytoscreening for the early diagnosis of endometrial carcinoma has thus far been hampered by the low diagnostic accuracy of current cytopathology. In this study we have analyzed the reactivity of the two monoclonal antibodies, AR-3 and B72.3, recognizing two distinct glycosylated high molecular weight carcinoma associated antigens on histological specimens from normal, hyperplastic, and transformed endometrium with the aim of establishing their diagnostic potential. Because women with a high risk of endometrial cancer are frequently postmenopausal, where normal endometrium is characterized by atrophy and cystic glandular hyperplasia, the following findings were of interest. Both antibodies reacted with variable and apical staining patterns with a minority of specimens of normal cycling endometrium from premenopausal women. However, they were constantly negative when tested on normal atrophic postmenopausal endometrium, and only monoclonal antibody AR-3 occasionally stained glandular cystic hyperplasia. By contrast, lesions with atypical hyperplasia, which represent a preneoplastic condition, were stained by both antibodies in 89 or 67% of the cases depending on the monoclonal antibody used (100% if used in combination). Furthermore, 98% of the endometrial carcinomas tested were found to react with the combination of two monoclonal antibodies. If these findings are confirmed in a multicentric study, the use of the two reagents could be a valuable adjunct in the cytodiagnosis of endometrial cancer, especially in providing a guideline to selecting patients for endometrial curettage and additional diagnostic procedures.

Fibrogenesis and biosynthesis of elastin in cartilage.

This study presents direct evidence that dissociated chondroblasts from rabbit ear cartilage grown in vitro are capable of synthesizing insoluble elastin. Ultrastructural examination indicated that at an early stage of tissue development, elastogenesis is initiated producing a form of primary fibrils which later condense into an electron dense amorphous material which, unlike other elastin-containing tissues, is heavily stained by metal cations and lacks peripheral microfibrils. Native elastic fibrils and the mature elastic fiber bundles are both susceptible to elastase digestion. Transmission electronmicroscopy demonstrated the presence of many intracellular filaments all showing a substructural organization and localized in close proximity to the nucleus. Their possible contractile nature is discussed. Amino acid analysis of cartilage elastin and of the elastin synthesized in vitro revealed a close chemical similarity between the two molecules. Ultrastructural analysis of the in vitro elastin demonstrated a substructural organization quite similar to that of the elastin observed in an in vivo system.

Role of hyaluronic acid in the in vivo aggregation of cartilage proteoglycans.

From studies of isolated cartilage proteoglycans in solution it has been inferred that they occur in the tissue as aggregates of high molecular weight which consist of proteoglycan monomers, hyaluronic acid and specific link proteins. The present investigation provides direct evidence for the existence of hyaluronic acid-containing aggregates in vivo, as indicated by the following observations: Treatment of sections of coastal cartilage from newborn rabbits with Streptomyces hyaluronidase led to complete disappearance of the electron dense granules, which have been previously identified as chondroitin sulfate proteglycans, from the extracellular matrix. Similar results were obtained on digestion with leech hyaluronidase which, like the Streptomyces enzyme, specifically degrades hyaluronic acid. Proteoglycan aggregation occurs not only in the extracellular compartment but intracellularly as well, since a portion of the hyaluronidase-senstive, electron dense proteoglycan granules are found in intracellular vesicles. It is concluded that the ability of proteoglycan monomers to form aggregates is a true reflection of the in vivo organization of these molecules and that aggregate formation is an important factor in the maintenance of the normal physiological function of cartilage tissue.