Parvovirus B19 detection analysis in thyroid tissue paired samples: an observational study from a tertiary surgical oncology department.

PURPOSE: In the present study, we sought to investigate the presence of Parvovirus B19 in both abnormal and normal adjacent thyroid tissue specimens after total thyroidectomy as well as the extent that this phenomenon occurs in a population group referred to a tertiary surgical oncology department. METHODS: We detected Parvovirus B19 by Real-Time PCR in both abnormal and normal adjacent thyroid tissue specimens from 41 patients who underwent total thyroidectomy for thyroid disease (cancerous or benign). Hashimoto's thyroiditis, thyroid gland weight, maximum size of the predominant thyroid nodule as well as sex and age of the patients were also evaluated in respect to the Parvovirus B19 presence. RESULTS: Parvovirus B19 virus genome was detected in 21/41 (51.2%) patients in at least one of the paired thyroid tissue samples. No statistically significant difference was noted regarding the sex, age, postoperative diagnosis, thyroid weight and maximum nodule diameter and presence of multifocal disease. The correlation between the incidence of Hashimoto thyroiditis and absence of Parvovirus B19 genome was statistically significant. CONCLUSION: Our findings showed high prevalence of Parvovirus B19 DNA in thyroid tissue disease in the population examined. Its actual role of the virus and its potential implication in the development or progression of thyroid diseases remain to be elucidated. Larger cohort studies are needed in order to validate a quasi-mutually exclusive role of Hashimoto's thyroiditis and Parvovirus B19 presence in thyroid disease in terms of geographical distribution.

Recommendations for enterovirus diagnostics and characterisation within and beyond Europe.

Enteroviruses (EV) can cause severe neurological and respiratory infections, and occasionally lead to devastating outbreaks as previously demonstrated with EV-A71 and EV-D68 in Europe. However, these infections are still often underdiagnosed and EV typing data is not currently collected at European level. In order to improve EV diagnostics, collate data on severe EV infections and monitor the circulation of EV types, we have established European non-polio enterovirus network (ENPEN). First task of this cross-border network has been to ensure prompt and adequate diagnosis of these infections in Europe, and hence we present recommendations for non-polio EV detection and typing based on the consensus view of this multidisciplinary team including experts from over 20 European countries. We recommend that respiratory and stool samples in addition to cerebrospinal fluid (CSF) and blood samples are submitted for EV testing from patients with suspected neurological infections. This is vital since viruses like EV-D68 are rarely detectable in CSF or stool samples. Furthermore, reverse transcriptase PCR (RT-PCR) targeting the 5'noncoding regions (5'NCR) should be used for diagnosis of EVs due to their sensitivity, specificity and short turnaround time. Sequencing of the VP1 capsid protein gene is recommended for EV typing; EV typing cannot be based on the 5'NCR sequences due to frequent recombination events and should not rely on virus isolation. Effective and standardized laboratory diagnostics and characterisation of circulating virus strains are the first step towards effective and continuous surveillance activities, which in turn will be used to provide better estimation on EV disease burden.

Non-invasive detection of HPV DNA in exfoliative samples from ophthalmic pterygium: a feasibility study.

PURPOSE: The evaluation of a non-invasive detection method for human papilloma virus (HPV) in ophthalmic pterygia. METHODS: Cotton swab samples and corresponding tissue specimens were collected from 21 ophthalmic pterygia of 21 patients. HPV detection and typing were performed by real-time PCR. Discrepancies in HPV detection between swab and tissue samples as well as clinical correlations of findings were examined. RESULTS: HPV DNA was detected in 9 (42.86%) tissue specimens and 8 (38.09%) respective swab specimens. HPV genotypes 33, 39, 45, 56, 59 and 66 were identified in the examined specimens, while more than one strain's HPV type was detected in 2 specimens. HPV presence was significantly correlated with the female gender whereas other clinical associations were not statistically significant. CONCLUSIONS: Findings imply that PCR-mediated HPV detection and typing in exfoliative swab specimens may be employed as a non-invasive diagnostic tool in the management of ophthalmic pterygia.

Herpes and polyoma family viruses in thyroid cancer.

Thyroid cancer is considered the most common malignancy that affects the endocrine system. Generally, thyroid cancer derives from follicular epithelial cells, and thyroid cancer is divided into well-differentiated papillary (80% of cases) and follicular (15% of cases) carcinoma. Follicular thyroid cancer is further divided into the conventional and oncocytic (Hürthle cell) type, poorly differentiated carcinoma and anaplastic carcinoma. Both poorly differentiated and anaplastic carcinoma can arise either de novo, or secondarily from papillary and follicular thyroid cancer. The incidence of thyroid cancer has significantly increased for both males and females of all ages, particularly for females between 55-64 years of age, from 1999 through 2008. The increased rates refer to tumors of all stages, though they were mostly noted in localized disease. Recently, viruses have been implicated in the direct regulation of epithelial-mesenchymal transition (EMT) and the development of metastases. More specifically, Epstein-Barr virus (EBV) proteins may potentially lead to the development of metastasis through the regulation of the metastasis suppressor, Nm23, and the control of Twist expression. The significant enhancement of the metastatic potential, through the induction of angiogenesis and changes to the tumor microenvironment, subsequent to viral infection, has been documented, while EMT also contributes to cancer cell permissiveness to viruses. A number of viruses have been identified to be associated with carcinogenesis, and these include lymphotropic herpesviruses, namely EBV and Kaposi's sarcoma-associated herpesvirus [KSHV, also known as human herpesvirus type 8 (HHV8)]; two hepatitis viruses, hepatitis B virus (HBV) and hepatitis C virus (HCV); human papillomaviruses (HPVs); human T cell lymphoma virus (HTLV); and a new polyomavirus, Merkel cell polyomavirus identified in 2008. In this review, we examined the association between thyroid cancer and two oncogenic virus families, the herpes and polyoma family viruses, and we discuss their potential role as causative agents in thyroid carcinogenesis.

Investigation of BK virus, Epstein-Barr virus and human papillomavirus sequences in postoperative thyroid gland specimens.

BACKGROUND: Although recent evidence has implicated viruses in the regulation of epithelial-to-mesenchymal transition and tumor progression, little is known regarding viral infections in thyroid malignancies. Thus the aim of this study was to detect sequences of 3 potentially oncogenic viruses - BK virus (BKV), Epstein-Barr virus (EBV) and human papillomavirus (HPV) - in a series of postoperative thyroid gland specimens. METHODS: Thirty patients with thyroid nodules who underwent surgery for thyroid disease within a 3-year period were enrolled. Both nodular and adjacent normal thyroid tissue was surgically excised from each patient. Viral gene sequences of BKV (VP1), EBV (LMP1, EBNA2 and EBER1) and HPV were amplified by PCR. The PCR results were confirmed by direct sequencing analysis. RESULTS: VP1 gene sequences were detected in 60% (18/30) of thyroid cancer or multinodular hyperplasia lesions compared with in 43.3% (13/30) of adjacent normal thyroid tissue specimens. Fifteen of thirty (50%) of thyroid cancer or multinodular hyperplasia samples revealed LMP1 sequences compared with 46.7% (14/30) of corresponding normal thyroid tissues. EBNA2 gene sequences were detected in 90% (27/30) of thyroid cancer or multinodular hyperplasia samples, compared with 90% (27/30) of adjacent normal thyroid tissue specimens. All samples were negative for EBER1 sequences, while HPV DNA was not detected in either nodular or normal thyroid tissue. CONCLUSIONS: This study suggests that BKV and EBV "infection" is an early event, occurring within normal tissue. Our findings do not show a clear role for the viruses examined, instead they suggest an "endemicity" pattern rather than a causal effect.

Detection of human papillomavirus (HPV) DNA prevalence and p53 codon 72 (Arg72Pro) polymorphism in prostate cancer in a Greek group of patients.

Prostate cancer is the most common neoplasm found in males and the second most frequent cause of cancer-related mortality in males in Greece. Among other pathogens, the detection frequency of human papillomavirus (HPV) has been found to be significantly increased in tumor tissues among patients with sexually transmitted diseases (STDs), depending on the geographical distribution of each population studied. The present study focused on the detection of HPV and the distribution of Arg72Pro p53 polymorphism in a cohort of healthy individuals, as well as prostate cancer patients. We investigated the presence of HPV in 50 paraffin-embedded prostate cancer tissues, as well as in 30 physiological tissue samples from healthy individuals by real-time PCR. Furthermore, the same group of patients was also screened for the presence of the Arg72Pro polymorphism of the p53 gene, a p53 polymorphism related to HPV. Out of the 30 control samples, only 1 was found positive for HPV (3.33 %). On the contrary, HPV DNA was detected in 8 out of the total 50 samples (16 %) in the prostate cancer samples. The distribution of the three genotypes, Arg/Arg, Arg/Pro, and Pro/Pro, was 69.6, 21.7, and 8.7 % in the cancer patients and 75.0, 17.86, and 7.14 % in healthy controls, respectively. No statistically significant association was observed between the HPV presence and the age, stage, p53 polymorphism status at codon 72, or PSA. The increased prevalence of HPV detected in the prostate cancer tissues is in agreement with that reported in previous studies, further supporting the association of HPV infection and prostate cancer.

Detection and genotype analysis of human papillomavirus in non-small cell lung cancer patients.

Although the role of human papillomavirus (HPV) in the development of uterine cervical cancer is well established, the role of HPV in lung carcinogenesis remains controversial. The detection rates of HPV DNA are subject to a wide variation from 0 to 100%. This is partly influenced by the detection techniques employed. To elucidate the impact of HPV infection on lung parenchyma, we analyzed 100 non-small cell lung cancer (NSCLC) specimens (39 squamous cell carcinomas, 50 adenocarcinomas, 5 samples with characteristics of both squamous cell and adenocarcinoma, 5 undifferentiated and 1 large cell carcinoma) from the region of Crete, Greece. Sixteen non-cancerous samples served as the negative controls. DNA was extracted from 100 paraffin-embedded tissue sections obtained from NSCLC patients. The specimens were examined for the detection of HPV DNA by Real-Time PCR using GP5+/GP6+ primers. Furthermore, the HPV-positive samples were subjected to genotyping. In contrast to the absence of viral genomes in the control samples, HPV DNA was detected in 19 NSCLC specimens (19%). In particular, 4 squamous cell carcinomas, 12 adenocarcinomas, 1 sample with characteristics of both squamous cell and adenocarcinoma, and 2 undifferentiated samples were HPV-positive. The distribution of HPV genotypes was as follows: HPV 16: eight cases (42.1%); HPV 11: three cases (15.8%); HPV 6: one case (5.2%); HPV 59: one case (5.2%); HPV 33: two cases (10.5%); HPV 31: two cases (10.5%) and HPV 18: two cases (10.5%). The presence of HPV in the tumor samples provides evidence of the potential role of HPV in NSCLC and strongly argues for additional research on this issue.

Secondary rhinoplasty fixations with hyaluronic acid.

The management of nasal deformities especially after rhinoplasty is a challenge. Postsurgical edema may last 6-8 months, causing aesthetic irregularities and nose deformities. The aim of this study is to present the correction of minor nose deformities secondary to rhinoplasty using hyaluronic acid subdermal injections. Eleven patients were treated between 2009 and 2011 with subdermal injections of hyaluronic acid (24 mg/mL) with 0.3% lidocaine (Juvederm, Allergan, Pringy-France) at the 1-month follow-up visit. The volume of hyaluronic acid injected varied from 0.4 to 1 mL according to the deformity. Injections were aimed to correct minor surface irregularities and to provide aesthetic symmetry. These patients were followed for at least 12 months postoperatively. Irregularities were aesthetically corrected immediately after hyaluronic acid injections. No complications were reported with the exception of minor swelling that resolved within 1 week. Esthetic correction was achieved in all patients as determined by the surgeon as well as by overall patient's satisfaction. Our 1-year follow-up data suggest that hyaluronic acid absorption is slow enough to provide the necessary time for postsurgical edema resorption. Rhinoplasty is among the most commonly used procedures for aesthetic improvement in men and women. However, achievement of the final outcome may take several months due to the induced postsurgical edema. Subdermal hyaluronic acid injections can provide temporary correction of these nose irregularities. Our data suggest that subdermal hyaluronic acid injections may provide immediate and long-lasting correction of these minor deformities. As a result, the aesthetic outcome is achieved and maintained throughout the postsurgical course of edema decompression.

Expression analysis of B-Raf oncogene in V600E-negative benign and malignant tumors of the thyroid gland: correlation with late disease onset.

B-Raf, a member of the Raf serine/threonine kinase family, is an intermediate molecule in the mitogen-activated protein kinase pathway, which relays extracellular signals from the cell membrane to the nucleus via a cascade of phosphorylation events, ultimately promoting cancer development. This pathway is usually activated in human neoplasias. The purpose of this study was to investigate the role of B-Raf in thyroid pathology. We scanned for the presence of mutations at codon 600 (V → E) of the B-Raf gene, using a PCR-RFLP assay. In tumors with no mutation (32 benign and malignant thyroid tumors) and in their adjacent normal tissue, we measured the expression levels of B-Raf gene, using a quantitative real-time PCR (qPCR) assay. B-Raf expression in V600E-negative tumors deviated from the normal pattern, since it was overexpressed in 42 % of benign samples and downregulated in 54 % of malignant specimens. Hashimoto's thyroiditis also seemed to play an important role, since benign specimens with Hashimoto's thyroiditis had a 2.2-fold higher B-Raf expression than samples without thyroiditis (1.71 ± 0.63 vs. 0.78 ± 0.13). Statistical analysis revealed that B-Raf deregulation postponed disease onset by more than 10 years in both benign and malignant thyroid (benign: 55.6 ± 3.9 vs. 45.3 ± 3.3, p = 0.049; malignant: 52.2 ± 3.5 vs. 33.0 ± 7.9, p = 0.020). From the above results, we deduce that in the absence of mutation activation, B-Raf overexpression or downregulation is a protective event, since it delays the development of both malignant and benign thyroid tumors.