Human papillomavirus (HPV) infection is the cause of cervical cancer. Integration of HPV-16 DNA in cervical cells is considered to be a key event in the progression towards invasive cancer, but little is known about this event in anal carcinogenesis. The integration could be a useful biomarker for cancer progression. Optimized assays are needed to determine the value of real-time detection of HPV integration in longitudinal studies, and this approach is only possible with a high-throughput assay. The aim of this study was to develop a new multiplex real-time PCR assay based on simultaneous amplification of the E2 and E6 HPV open reading frames (ORFs) in order to assess the physical status (episomal and/or integrated) of HPV-16 in anal cells of HIV-positive men. The comparative threshold (Ct) cycle values for E2 and E6 obtained for SiHA cells and artificial mixtures of episomal and integrated DNA were as expected: similar Ct for episomal forms and absence of E2 amplification for integrated forms. The multiplex real-time PCR was tested in 77 consecutive samples from individual HIV-infected patients with HPV-16 anal infection. The integration of HPV-16 was detected in 25 (32%) patients: 23 as mixed (episomal and integrated) and two as completed integrated forms. The integration occurs in the early stage of anal lesions and was associated with the severity of the lesions (p 0.004). The multiplex real-time PCR assay developed in the course of this study was shown to be a simple, sensitive, specific and inexpensive technique which may be applied routinely to detect HPV-16 integration.

No abstract

HPV-16 infection and its integration in anal cells were highly prevalent in HIV-infected men. The assessment of HPV-16 integration rather than HPV-infection could be a good biomarker for predicting anal precancerous lesions in HIV-positive men.

No abstract

The prevalence and distribution of principal HPV types involved in the carcinogenesis process of the cervix were similar in HIV-infected and noninfected women, although a tendency toward a lower HPV-16 and a higher HPV-18 prevalence in invasive cervical carcinoma was detected in HIV-positive women. Similar percentage of HPV-16 and HPV-18 viral integration was found in formaldehyde-fixed paraffin-embedded specimens of cervical cancer regardless of the HIV infection status.

Objective: Detection of high-risk human papillomavirus types (HPV) infection is an important tool in the screening of cervical cancer and triage of cytological abnormalities. The different techniques for detection of this cancer need to be contrasted and validated for use in population screening. Material and Methods: Cervical cell samples were collected from 166 women attending a dermatology clinic in Oviedo (Spain). We evaluated the performance of three different assays for VPH detection. The methods utilized were 1) In-house PCR-EIA using L1 consensus primers MY09/ MY11, 2) A PCR-reverse line blot hybridization (PCR-LBH) that uses L1 consensus PGMY primers. 3) Hybrid Capture 2. All assays were performed blinded. The kappa statistic was used to test for global agreement between assay pairs. Results: HPV DNA was detected in 24,7%, 25,3% and 29,5% of the women, respective to the assay. The overall agreement between the in-house PCR, PCR-LBH and HC2 was (73.5%) with all kappa values between assay pairs exceeding 0.56 (p<0.001). Conclusion: The three HPV assays were equally accurate in estimating high-risk HPV prevalence and HPV-related lesions. The method for HPV detection must be decided depending on the goals of the search (screening, follow-up or molecular studies).

Background Cetuximab label indication includes treatment of epidermal growth factor receptor (EGFR)-expressing, KRAS wild-type metastatic colorectal cancer in several possible ways: combination with irinotecan-based chemotherapy, first-line in combination with FOLFOX and as a single agent after oxaliplatin- and irinotecan-based treatment failure in irinotecan-intolerant patients. In our hospital, a multidisciplinary team drawn from the Oncology and Pharmacy services has established a consensus for the rational use of cetuximab as first or second-line agent in association with other chemotherapeutic agents and as monotherapy in third-line treatment after the failure of oxaliplatin and irinotecan-based treatment. Purpose To verify the relevance of cetuximab prescription to the local protocol and cheque the label indications for cetuximab in our hospital. Materials and Methods A retrospective study of patients diagnosed with metastatic colorectal cancer between 2006–2012 with available KRAS status. Patients were followed up for a minimum of three months after diagnosis. ResultsTwenty-six patients were collected (mean age: 62.2 ± 12.6 years; 53.8% male). KRAS mutation was negative in 42.3% (11/26) patients and therefore they were eligible for treatment with cetuximab. Five out of those 11 patients underwent cetuximab treatment (5/11; 45.5%): three associated with oxaliplatin in first-line treatment, one associated with irinotecan in second-line treatment and one as monotherapy in second-line treatment. Four out of these 5 prescriptions of cetuximab were in accordance to our local protocol and label (4/5; 80.0%). One prescription was not in accordance with either the local protocol or the cetuximab label; due to this the patient was treated with oral capecitabine as first-line and cetuximab monotherapy as second-line treatment. Three KRAS-negative patients (3/11; 27.3%) are currently in treatment with irinotecan as second-line therapy. Three KRAS-negative patients were lost to follow-up after undergoing second-line treatment not known to contain a cetuximab prescription (3/11; 27.3%). Fifteen patients positive for KRAS mutation (15/26; 57.7%) were not treated with cetuximab. Conclusions Ninety-five percent of cetuximab prescriptions in our hospital are in accordance with the established local protocol and the cetuximab label (19/20). No conflict of interest.

IntroductionOpioid addiction is a serious health/social problem, associated with high morbidity and mortality.Several gene polymorphisms on the mu-opioid receptor gene(OPRM1), which plays an important role in reward system, have been related to opioid dependence (A118G, C17T, C2044A). A118G is the most studied and probably the most promising biomarker of better response in these patients, despite discrepancies has been manifested even in studies conducted on the same ethnicity.Objectives and aimsDescription of A118G, C17T and C2044A allele frequencies in an opioid-dependent population. Evaluation of the association of A118G gene polymorphism with opioid dependence.MethodsCase-control Study.Case group: 16 patients with opioid addiction, included in a Heroin Prescription Program in Andalusia, based on the protocolized individual prescription of diacetylmorphine. Control group: 32 non opioid-dependent subjects.Genotyping of A118G, C17T and C2044A was performed by Polymerase Chain Reaction and Restriction Fragment Length Polymorphism.ResultsCase group: 12 patients were AA homozygous (12/16;75%) and 4 patients were heterozygous AG (4/12;25%) for A118G. All patients were homozygous CC for C17T and C2044A (16/16;100%). The distribution of the genotype frequencies of OPRM1 gene polymorphisms in the case series were not statistically different from those reported for European populations in HapMap for A118G (p=0.6418) and the GENO PANEL for C17T. Control group:19 patients were AA homozygous(19/32; 59.4%) and 13 patients were heterozygous AG (13/32;40.6%) for A118G. This polymorphism was not associated to opioid addiction (p=0.3503).ConclusionsDistribution of genotype frequencies in opioid dependants corresponded to specific frequencies from European population for A118G and C17T polymorphisms. OPRM1 gene polymorphisms were not associated to opioid addiction in this population.