Human papillomavirus viral load: a possible marker for cervical disease in HIV-infected women

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60PV Related Lesions in HIV Positive Subjects on HAART: Study of Viral Markers of Cervical Neoplasia Progression

Methods HIV pos women (N = 410) were followed since 1995 in a longitudinal study. Each one undergo periodic (6–12 months) colposcopy, PAP smear, biopsy if needed, and cervical sampling for HPV testing. HIV related parameters (CD4, HIV-RNA, ART) are recorded and related to cervical disease. HPV typing is performed by reverse hybridisation assays, viral load by in-house real time PCR and viral expression by E6/E7 mRNA detection. The Mann-Whitney rank sum test for non-parametric data and the association between discrete variables by Chi-square test of Fisher exact test were applied

Comparative Analysis of Five Multiplex RT-PCR Assays in the Screening of SARS-CoV-2 Variants

The rapid and presumptive detection of SARS-CoV-2 variants may be performed using multiplex RT-PCR assays. The aim of this study was to evaluate the diagnostic performance of five qualitative RT-PCR tests as compared with next-generation sequencing (NGS). We retrospectively examined a multi-variant panel (n = 72) of SARS-CoV-2-positive nasopharyngeal swabs categorized as variants of concern (Alpha, Beta, Gamma and Delta), variants under monitoring (Iota and Kappa) and wild-type strains circulating in Liguria (Italy) from January to August 2021. First, NGS libraries of study samples were prepared and mapped to the reference genome. Then, specimens were screened for the detection of L452R, W152C, K417T, K417N, E484Q, E484K and N501Y mutations using the SARS-CoV-2 Variants II Assay Allplex, UltraGene Assay SARS-CoV-2 452R & 484K & 484Q Mutations V1, COVID-19 Ultra Variant Catcher, SARS-CoV-2 Extended ELITe MGB and Simplexa SARS-CoV-2 Variants Direct. The overall accuracy of these assays ranged from 96.9% to 100%. Specificity and sensitivity were 100% and 96-100%, respectively. We highly recommend the use of these assays as second-level tests in the routine workflow of SARS-CoV-2 laboratory diagnostics, as they are accurate, user friendly, low cost, may identify specific mutations in about 2-3 h and, therefore, optimize the surveillance of SARS-CoV-2 variants

HIV, human papillomavirus, and cervical neoplasia and cancer in the era of highly active antiretroviral therapy

The objective of this study was to review the literature on the epidemiological association between human papillomavirus (HPV), HIV, and cervical neoplasia, and the impact of highly active antiretroviral therapy (HAART) on this association. MEDLINE was searched using the terms 'human papillomavirus, 'HPV, 'HIV', 'cervix, 'neoplasm', and 'antiretroviral' to identify articles published before December 2006. HIV-infection was strongly associated with a higher prevalence, incidence, and persistence of HPV infection and correlated with prevalence, incidence, persistence, and progression of squamous intraepithelial lesions. The association between HIV and invasive cervical carcinoma has been more difficult to establish, but is now fully recognized. HAART seems to have little, if any, beneficial effect on the natural history of intraepithelial lesions in HIV-positive women. Despite this fact, HAART, does increase the life expectancy of HIV-positive women. Therefore, it remains important to closely monitor HPV-related disease in women with HIV who are receiving HAART, particularly in regions of the world where cervical screening is not available routinely

CD4+ T cells against human papillomavirus-18 E7 in patients with high-grade cervical lesions associate with the absence of the virus in the cervix

Cervical neoplastic lesions are associated with infection by high-risk human papilloma-viruses (HPV). The two genotypes most frequently found in the lesions are HPV-16 and HPV-18 with a prevalence of 50–60% and 15–18%, respectively. The E6 and E7 viral oncoproteins are involved in the transformation process and represent foreign antigens for the host. We previously reported that anti-HPV-18 E6 CD4+ T cells are present in patients with high-grade HPV-18-expressing cervical lesions but also in 50% of the total consecutive patients tested, independently of the HPV type carried. These results indicated that HPV-18 E6 is immunogenic and suggested that all responsive patients, irrespective of the HPV expressed, had encountered HPV-18 and cleared the infection. Here, we investigated anti-HPV-18 E7 CD4+ T-cell immunity in a cohort of 23 HPV-18 E6-responsive patients. We found that, although E7-specific CD4+ T cells were present in all women, a robust T helper type (Th1)/Th2 type response against E7 was associated with HPV-18-negative status, suggesting that indeed these patients might have cleared the virus. In agreement with this hypothesis, we found strong anti-E7 CD4+ T-cell immunity in 20% of 24 healthy donors without evidence of disease. In contrast, a robust Th1/Th2 type response against E6 but not E7 correlated with a lack of disease relapse and/or infection recurrence but did not discriminate between HPV-18-positive and HPV-18-negative patients. Collectively, our data suggest different roles for anti-HPV-18 E6 and E7 CD4+ T cells in anti-viral and anti-tumour immunity

Unexpected elevated free thyroid hormones in pregnancy

Background: The use of thyrotropin and free thyroid hormone assays to evaluate thyroid function is widespread, but in some situations the results are inconsistent with the patient's thyroid status. Summary: A 35-year-old woman with a known diagnosis of chronic autoimmune thyroiditis was referred to the authors' clinic at week 26 of her second pregnancy. The patient was clinically euthyroid. Consistent with this, her serum thyrotropin (TSH) was normal (0.79 mIU/L), but she had elevated free thyroid hormones-free triiodothyronine (fT3) and free thyroxine (fT4)-As determined by a one-step chemiluminescent assay. The patient was taking levothyroxine replacement therapy (125 lg/day), and the dose was confirmed. Previous blood tests showed concordance between TSHand free thyroid hormone values. The patient was followed up throughout gestation and at 12months postpartum. During gestation, her free thyroid hormones remained high using one-step methods, while the total thyroid hormone concentration values were within the reference range, in agreement with the TSH values. Postpartum fT4 and fT3 values returned progressively to normality, in agreement with the TSH values. The presence of circulating thyroid hormone autoantibodies (THAb) was hypothesized, which are known to interfere, although to a variable extent, with thyroid hormone one-step assays. Using stored frozen sera, this hypothesis was confirmed indirectly by measuring normal levels of fT3 and fT4 with a two-step method, and directly by demonstrating THAb against the two hormones. Conclusion: Despite their relative rarity, circulating THAb may be suspected when laboratory data are not consistent and contrast with the clinical picture. To the authors' knowledge, no previous case of transient appearance of THAb in pregnancy has been described

The prevalence of human papillomavirus infection in Mombasa, Kenya

Objectives A human papillomavirus (HPV) prevalence survey was done in Mombasa, Kenya, to improve the knowledge of HPV prevalence and genotype distribution in sub-Saharan African countries overall, and in women of different ages. Methods HPV prevalence was assessed using PCR in women older than 15 years attending family planning and mother-child care services. Results Among 496 women, HPV prevalence was high (42.3%; 95% CI: 37.9-46.8; world age-standardized). Moreover, 46% of HPV-positive women harbored multiple-type infections. The most common types were HPV58 (10.5% of women), HPV16 (7.7%), HPV53 (6.7%), HPV18 (4.6%), and HPV6 (4.4%), and the prevalence of any high-risk HPV type was 28.8%. HPV prevalence was elevated among all age-groups (range 36.4-45.7%). Independent associations with HPV positivity were found for being in a polygamous marriage (OR = 1.7) and lifetime number of sexual partners (OR for >= 3 vs. 1 = 1.5), although they were of only borderline statistical significance. Conclusions These findings differ from other world regions, showing a high HPV burden in all age-groups with a high proportion of multiple-type infections. Our data strengthen the urgency of HPV vaccination in Kenya but also highlight the elevated number of women who would have positive results in an HPV-based screening program in the country

Titolazioni dei preparati galenici oleosi a base di cannabis in Regione Liguria: progetto sperimentale dei laboratori di riferimento regionale

Analysis of cannabinoids concentration in cannabis oil galenic preparations in the Liguria Region: experimental project of the regional reference laboratories. Introduction: the medical use of cannabis is increasingly being applied in the treatment and support of several diseases and syndromes. In the Liguria Region, olive oil galenic preparations are mainly prepared by hospital pharmacies, according to common standard procedures. The preparations must be analyzed in order to establish the concentration of the two main active compounds (delta-9 tetrahydrocannabinol, THC and cannabidiol, CBD) thus allowing the correct setting of the therapeutic prescription. Liguria Region is at the forefront in the use of medical cannabis with a high number of patients treated (>1000). The aim of this work is to describe the organization of the titration activity centralized at the two regional reference laboratories (Central Laboratory of Analyses of Giannina Gaslini Institute, Genova and Toxicology Laboratory of Sarzana, La Spezia), coordinated by the inter-hospital department (DIAR) of the Laboratories Area. Methods: the phases of the analytical process (pre-analytical, analytical and post-analytical) have been identified and described. The analysis of the workflow has been carried out including the methods to prepare cannabis oil in the pharmacies, the intervals and production volumes, the medical-legal handling requirements and operational responsibilities. The definition of the pre-analytical phase foresees the methods of packaging, transport and recording of the samples and related responsibilities. Results: the analytical phase included the development and validation of the analytical method Ultra High Performance Liquid Chromatography coupled to tandem mass spectrometry, (UHPLC-MS/MS) in the two laboratories, with common procedures and the comparison of results conducted both on reference material and real samples of olive oil galenic preparations. The definition of the post-analytical phase included the reporting procedures. Discussion: the experimental phase has been concluded at the end of 2019 and the implementation phase of the project has started in march 2020