903 Protein Saver cards: the best alternative for dried blood spot storage at room temperature for HCV RNA.

Hepatitis C virus (HCV) infection remains a global health problem, detected only in the early stages by molecular tests. Molecular tests detect HCV RNA, which is very prone to degradation by ribonucleases, reason why blood samples must be transported and stored at - 20 °C, or even - 70 °C for long-term storage. Flinders Technology Associates (FTA) cards are a useful sampling collecting device for dry blood spot (DBS) storage, especially for low and middle-income countries (LMIC). In this study, we analyzed viral HCV RNA integrity for long-term storage at room temperature compared to - 20 °C using two different types of cards for DBS: FTA Classic and 903 Protein Saver cards. For this purpose, DBS were prepared on these cards using blood or plasma samples from HCV infected patients, and samples were analysed by conventional RT-PCR. Our results showed that 903 Protein Saver cards are the best and cheapest alternative for DBS storage at room temperature. In these conditions, we found that HCV RNA integrity lasted for up to 9 months.

p38γ and p38δ regulate postnatal cardiac metabolism through glycogen synthase 1.

During the first weeks of postnatal heart development, cardiomyocytes undergo a major adaptive metabolic shift from glycolytic energy production to fatty acid oxidation. This metabolic change is contemporaneous to the up-regulation and activation of the p38γ and p38δ stress-activated protein kinases in the heart. We demonstrate that p38γ/δ contribute to the early postnatal cardiac metabolic switch through inhibitory phosphorylation of glycogen synthase 1 (GYS1) and glycogen metabolism inactivation. Premature induction of p38γ/δ activation in cardiomyocytes of newborn mice results in an early GYS1 phosphorylation and inhibition of cardiac glycogen production, triggering an early metabolic shift that induces a deficit in cardiomyocyte fuel supply, leading to whole-body metabolic deregulation and maladaptive cardiac pathogenesis. Notably, the adverse effects of forced premature cardiac p38γ/δ activation in neonate mice are prevented by maternal diet supplementation of fatty acids during pregnancy and lactation. These results suggest that diet interventions have a potential for treating human cardiac genetic diseases that affect heart metabolism.

Discovery of Diverse Natural Products as Inhibitors of SARS-CoV-2 Mpro Protease through Virtual Screening.

SARS-CoV-2 is a type of coronavirus responsible for the international outbreak of respiratory illness termed COVID-19 that forced the World Health Organization to declare a pandemic infectious disease situation of international concern at the beginning of 2020. The need for a swift response against COVID-19 prompted to consider different sources to identify bioactive compounds that can be used as therapeutic agents, including available drugs and natural products. Accordingly, this work reports the results of a virtual screening process aimed at identifying antiviral natural product inhibitors of the SARS-CoV-2 Mpro viral protease. For this purpose, ca. 2000 compounds of the Selleck database of Natural Compounds were the subject of an ensemble docking process targeting the Mpro protease. Molecules that showed binding to most of the protein conformations were retained for a further step that involved the computation of the binding free energy of the ligand-Mpro complex along a molecular dynamics trajectory. The compounds that showed a smooth binding free energy behavior were selected for in vitro testing. From the resulting set of compounds, five compounds exhibited an antiviral profile, and they are disclosed in the present work.

Analysis of the prevalence of human papillomavirus and abnormal anal cytology in women at risk.

Our aim was to analyze the association between anal human papillomavirus (HPV) infection and abnormal anal cytology in women with high-grade cervical intraepithelial neoplasia (CIN 2+). We also analysed what other risk factors might be significant. We carried out a prevalence study from April 2015 to March 2017 at La Paz University Hospital. Genotyping of HPV, anal cytology and high-resolution anoscopy were performed. Of 171 patients recruited, 53 cases (31%) were diagnosed as histological CIN 2+: there were no statistically significant differences in the prevalence of anal HPV (OR = 0.8), nor the prevalence of abnormal anal cytology (OR = 2.15, 95% CI 0.8-5.7) compared to women with CIN 1 or no cervical dysplasia. Immunosuppression (OR = 2.51, 95% CI 1-6.3, p < .05), cervical HPV (OR = 3.9, 95% CI 1.9-8.0, p < .01) and being older than 40 years old (p < .05) were also associated with anomalous anal results.Impact StatementWhat is already known on this subject? Anal HR-HPV and abnormal anal cytology may precede anal intraepithelial neoplasia (AIN): a premalignant lesion that may progress to anal cancer. It is known that there are four populations which present a higher risk of developing anal cancer compared to the general population: human immunodeficiency virus (HIV)-positive patients, other immunocompromised populations, men who have sex with men and women with a history of disease secondary to HPV infection.What do the results of this study add? This study allowed us to compare the prevalence of anal HPV and abnormal anal cytology in women with CIN 2+: it analysed whether these women already presented alterations in anal tests at the moment of the diagnosis of the preneoplastic cervical lesion. It also provides information for the management of the populations at a higher risk of developing anal cancer; specifically, the group of women with a prior history of HPV-associated anogenital disease.What are the implications of these findings for clinical practice and/or further research? Our findings improve the existing evidence on anal HPV infection and anal cytology on the least studied population at risk. Data could be useful for further research in order to clarify the role of anal screening in this population and standardise the clinical practice.

Lack of Survival Benefit of Para-Aortic Lymphadenectomy in Advanced Cervical Cancer.

INTRODUCTION: The presence of positive para-aortic lymph nodes in advanced cervical cancer remains the most important prognostic factor for survival and also defines the treatment. Our aim was to define the influence of staging para-aortic lymphadenectomy in patients' survival. MATERIAL AND METHODS: The medical records of 74 patients with advanced cervical cancer (FIGO IIB-IVA) were reviewed. In 31 patients (41.9%), the assessment of lymph nodes was performed with imaging test (group 1) and in 43 (58.1%) within a surgical staging para-aortic lymphadenectomy (group 2). We compared both groups according to stage of disease, treatment, progression-free survival (PFS), and overall survival (OS). RESULTS: The extended-field radiotherapy was performed in 44.2 and 19.4% of patients in surgical and imaging staging group, respectively (p = 0.045). The disease-free survival rate was 17.4 ± 17.4 months in group 1 and 14.4 ± 12.6 months in group 2 (p = 0.456). No differences in OS were found between these 2 groups (p = 0.676). CONCLUSIONS: Despite the higher diagnostic accuracy of surgical staging and the higher number of patient who received extended field radiotherapy, we did not find differences between the overall and PFS rates in both the studied groups. Further prospective study on a higher number of patients would be necessary.

Ausencia de lesión intraepitelial en piezas de conización / Related factors to absence of cervical intraepithelial neoplasia in the conization specimen

Resumen OBJETIVO: Investigar las posibles causas de los conos blancos, establecer estrategias para disminuir su incidencia y desarrollar protocolos de seguimiento. MATERIALES Y MÉTODOS: Estudio observacional, retrospectivo, de casos y controles. Se incluyeron las pacientes a quienes se efectuó una conización en el Hospital Universitario La Paz. Las variables analizadas más importantes fueron: anatomía patológica de la pieza y su relación con la biopsia y citologías previas, longitud del cono, presencia o no de artefacto y de cervicitis. Para el análisis estadístico se utilizaron: χ2, prueba exacta de Fisher, t de Student, U de Mann-Whitney, Kruskal-Wallis y Kolmogorov-Smirnov. RESULTADOS: Se integraron dos grupos: 371 conos positivos (85.9%) y 61 negativos (14.1%), con diferencias estadísticamente significativas en la citología, colposcopia y biopsia. Hubo mayor porcentaje de lesiones de menor grado en las pacientes con conización blanca. La longitud del cono fue menor en el grupo de análisis y en éste también se observó mayor porcentaje de cervicitis y artefactos. CONCLUSIONES: Las causas de lesión residual luego de una conización son variadas y difíciles de demostrar. Las pacientes con citología anómala e inflamación o atrofia deben recibir tratamiento para evitar falsos positivos y mejorar la técnica quirúrgica para impedir artefactos.

Abstract OBJECTIVES: to investigate the possible causes of the negative cones, to establish strategies to reduce their incidence and to develop monitoring protocols. MATERIALS AND METHODS: This is a retrospective observational cases and controls study of 432 conizations made in the Hospital Universitario La Paz (HULP) between 2013 and 2015. The most important analysed variables were the pathological anatomy of the piece and its relationship with the biopsy and previous cytology, the cone length, as well as the presence and artefact and cervicitis. The analysis it was used Chi - Square and Fisher´s test, T-Student, Mann Whitney U, Kruskal-Wallis and Kolmogorov- Smirnov. RESULTS: There are two groups: 371 positive (85,9%) and 61 negative cones (14,1%). We find statistically significant differences in the cytology, colposcopy and biopsy pre-conization, finding a major percentage of injuries of lesser degree in the patients with negative cone. The length of the cone was lower in the analysis group and in this we also observed a greater percentage of cervicitis and artefacts. CONCLUSIONS: The causes that make the remaining injury not appear after a diagnosed and/or therapeutic conization are a wide variety and difficult to prove. We should try to treat the patients with inflammation or atrophy to avoid false positives in the cytology and biopsy, improve the surgical technique to avoid artefacts and perform conservative management of low-risk injuries.

Surgical outcome of extraperitoneal paraaortic lymph node dissections compared with transperitoneal approach in gynecologic cancer patients.

STUDY OBJECTIVE: To evaluate the surgical outcome of extraperitoneal paraaortic lymph node dissection compared with the traditional transperitoneal approach. DESIGN: Retrospective review (Canadian Task Force classification III). SETTING: University hospital. PATIENTS: Women with gynecologic malignancies admitted to our hospital between 2007 and 2011 who underwent laparoscopic paraaortic lymphadenectomy. INTERVENTIONS: Indication, diagnosis, and outcome according to type of surgery were evaluated. MEASUREMENTS AND MAIN RESULTS: Of 47 patients who underwent laparoscopic paraaortic lymphadenectomy because of gynecologic indications, 28 patients underwent extraperitoneal paraaortic lymph node dissection and 19 underwent the same procedure via the classic transperitoneal technique. The most frequent indication for extraperitoneal lymph node dissection was cervical cancer (71.4%), and for the transperitoneal technique was endometrial cancer (47.4%). The mean (SD) duration of surgery was 211 (38) minutes in the transperitoneal approach group, and 173 (51) minutes in the extraperitoneal lymphadenectomy group (p = .009). No significant differences between groups were found in the number of lymph nodes removed (15 [5.9] nodes in the extraperitoneal group vs 17.4 [8.6] in the transperitoneal group; p = .25). However, a higher rate of positive nodes was observed in the extraperitoneal group than in the transperitoneal group (42.8% vs 36.2%, respectively [p = .001]), and a significantly shorter stay in the intensive care unit in the extraperitoneal group (0.59 [0.5] vs 1.1 [0.5] days, respectively; p = .02). No significant differences in complication rate were found between groups. CONCLUSIONS: Extraperitoneal paraaortic lymph node dissection is a minimally invasive procedure that is an excellent and safe approach to the paraaortic area, with a low complication rate, sufficient number of lymph nodes, and short hospital stay. It seems to be a good alternative to the classic transperitoneal approach.

[Laparoscopic removal of primary intestinal fibroid]. / Extirpación laparoscópica de mioma intestinal primario.

Uterine fibroids are the most frequent solid benign tumors of the female pelvis. The most frequent location is the uterine corpus; however, it has been also reported in cervical placement and broad ligament. Rarely it has been reported a case of intestinal fibroid after previous history of myomectomy. We report the case of a patient, with no surgical history of interest, who underwent a diagnostic laparoscopy due to suspicion of malignant abdominal mass. Ajejunal fibroid was found, which was removed via laparoscopy without complications. Laparoscopy may be considered in case of intestinal fibriod despite its rarity, due to the better postoperative recovery and cosmetic outcomes compared to laparotomy.