Proteomic snapshot of saliva samples predicts new pathways implicated in SARS-CoV-2 pathogenesis.

BACKGROUND: Information on the microbiome's human pathways and active members that can affect SARS-CoV-2 susceptibility and pathogenesis in the salivary proteome is very scarce. Here, we studied a unique collection of samples harvested from April to June 2020 from unvaccinated patients. METHODS: We compared 10 infected and hospitalized patients with severe (n = 5) and moderate (n = 5) coronavirus disease (COVID-19) with 10 uninfected individuals, including non-COVID-19 but susceptible individuals (n = 5) and non-COVID-19 and nonsusceptible healthcare workers with repeated high-risk exposures (n = 5). RESULTS: By performing high-throughput proteomic profiling in saliva samples, we detected 226 unique differentially expressed (DE) human proteins between groups (q-value ≤ 0.05) out of 3376 unambiguously identified proteins (false discovery rate ≤ 1%). Major differences were observed between the non-COVID-19 and nonsusceptible groups. Bioinformatics analysis of DE proteins revealed human proteomic signatures related to inflammatory responses, central cellular processes, and antiviral activity associated with the saliva of SARS-CoV-2-infected patients (p-value ≤ 0.0004). Discriminatory biomarker signatures from human saliva include cystatins, protective molecules present in the oral cavity, calprotectins, involved in cell cycle progression, and histones, related to nucleosome functions. The expression levels of two human proteins related to protein transport in the cytoplasm, DYNC1 (p-value, 0.0021) and MAPRE1 (p-value, 0.047), correlated with angiotensin-converting enzyme 2 (ACE2) plasma activity. Finally, the proteomes of microorganisms present in the saliva samples showed 4 main microbial functional features related to ribosome functioning that were overrepresented in the infected group. CONCLUSION: Our study explores potential candidates involved in pathways implicated in SARS-CoV-2 susceptibility, although further studies in larger cohorts will be necessary.

The microbiome as a biomarker of anal precancerous lesions in people with HIV.

PURPOSE OF REVIEW: Early detection and treatment of human papillomavirus (HPV)-related anal dysplasia in some high-risk groups can help anal cancer prevention, but new tools to improve diagnostic and risk assessment are needed. Here, we aim to discuss the evidence on the role of the microbiome as a potential biomarker for anal high-grade squamous intraepithelial lesions (HSILs) in people with HIV (PWH). RECENT FINDINGS: This review covers relevant studies on the links between the microbiome and HPV infection, cervical dysplasia/cancer, and anal HPV disease. It focuses on anal samples and precancerous lesions. SUMMARY: The review highlights the promising potential of the anal microbiome as a novel biomarker for precancerous lesions in people with HIV, while also discussing limitations and future research needs.

Implementation of a population-based cancer registry network in Mexico 2017-2020.

The growing cancer burden particularly among less developed countries requires local data to plan and evaluate cancer control measures. This article describes the development of a population-based cancer registry network (PBCRN) in Mexico that took place between 2017 and 2020 and present related data. The PBCRN, led by the National Cancer Institute (Incan), included nine registries representing 11.3% of the Mexican population. Definitions, coding, and operative processes were based on international standards. All cities were visited to set up local structure; personnel were hired by Incan and trained in basic cancer registration in Merida. A specific software was developed. Regular virtual meetings took place for data verification and quality control. Data collection included institutions of the public and private health system. Personnel included 34 registrars, nine local leaders, and 12 staff members at the Incan. A total of 13 517 cases were recorded between 2017-2020, 64% percent of them were among females. Breast cancer was the more frequent malignancy (23.3%), followed by digestive organs with (18.4%) and female genital cancers (13.5%). Childhood (0-14 years) and adolescents cancer represented 4.4% of the total new cancer cases. The network was suspended in 2020. The present effort lacked sustainability and data were only partial. However, the experience provides valuable insights to be considered for the renewed cancer registration efforts that are currently ongoing in Mexico.

Microbiome-derived cobalamin and succinyl-CoA as biomarkers for improved screening of anal cancer.

Human papillomavirus can cause preinvasive, high-grade squamous intraepithelial lesions (HSILs) as precursors to cancer in the anogenital area, and the microbiome is suggested to be a contributing factor. Men who have sex with men (MSM) living with human immunodeficiency virus (HIV) have a high risk of anal cancer, but current screening strategies for HSIL detection lack specificity. Here, we investigated the anal microbiome to improve HSIL screening. We enrolled participants living with HIV, divided into a discovery (n = 167) and validation cohort (n = 46), and who were predominantly (93.9%) cisgender MSM undergoing HSIL screening with high-resolution anoscopy and anal biopsies. We identified no microbiome composition signatures associated with HSILs, but elevated levels of microbiome-encoded proteins producing succinyl coenzyme A and cobalamin were significantly associated with HSILs in both cohorts. Measurement of these candidate biomarkers alone in anal cytobrushes outperformed anal cytology as a diagnostic indicator for HSILs, increasing the sensitivity from 91.2% to 96.6%, the specificity from 34.1% to 81.8%, and reclassifying 82% of false-positive results as true negatives. We propose that these two microbiome-derived biomarkers may improve the current strategy of anal cancer screening.

Novel Approaches in the Systemic Management of High-Risk Prostate Cancer.

Locally advanced prostate cancer comprises approximately 20% of new prostate cancer diagnoses. For these patients, international guidelines recommend treatment with radiotherapy (RT) to the prostate in combination with long-term (2-3 years) androgen deprivation therapy (ADT), or radical prostatectomy in combination with extended pelvic lymph node dissection (PLND) as another treatment option for selected patients as part of multimodal therapy. Improvements in overall survival with docetaxel or an androgen receptor signaling inhibitor have been achieved in patients with metastatic castration sensitive or castration resistant prostate cancer. However, the role of systemic therapy combinations for high risk and/or unfavorable prostate cancer is unclear. In this context, the aim of this review is to assess the current evidence for systemic treatment combinations as part of primary definitive therapy in patients with high-risk localized prostate cancer.

CD4/CD8 Ratio During Human Immunodeficiency Virus Treatment: Time for Routine Monitoring?

In the last decade, studies in persons with HIV (PWH) on antiretroviral therapy (ART) have shed light on the significance of persistently high CD8 counts and low CD4/CD8 ratios. A low CD4/CD8 ratio reflects increased immune activation and is associated with an increased risk of severe non-AIDS events. As a result, many clinicians now believe that the CD4/CD8 ratio can help in HIV monitoring, and many researchers now report it as an efficacy marker in interventional studies. However, the topic is more complex. Recent studies have not yielded unanimous conclusions on the ability of the CD4/CD8 ratio to predict adverse outcomes, and only some clinical guidelines recommend monitoring it. Knowledge gaps remain on the best cutoff points, associated clinical events, effects of treatments, and how the CD4/CD8 ratio could improve decision making in the clinic. Here, we critically review the literature, identify knowledge gaps, and discuss the role of the CD4/CD8 ratio as a marker for HIV monitoring.

The microbiota as a modulator of mucosal inflammation and HIV/HPV pathogenesis: From association to causation.

Although the microbiota has largely been associated with the pathogenesis of viral infections, most studies using omics techniques are correlational and hypothesis-generating. The mechanisms affecting the immune responses to viral infections are still being fully understood. Here we focus on the two most important sexually transmitted persistent viruses, HPV and HIV. Sophisticated omics techniques are boosting our ability to understand microbiota-pathogen-host interactions from a functional perspective by surveying the host and bacterial protein and metabolite production using systems biology approaches. However, while these strategies have allowed describing interaction networks to identify potential novel microbiota-associated biomarkers or therapeutic targets to prevent or treat infectious diseases, the analyses are typically based on highly dimensional datasets -thousands of features in small cohorts of patients-. As a result, we are far from getting to their clinical use. Here we provide a broad overview of how the microbiota influences the immune responses to HIV and HPV disease. Furthermore, we highlight experimental approaches to understand better the microbiota-host-virus interactions that might increase our potential to identify biomarkers and therapeutic agents with clinical applications.

Selective miRNA inhibition in CD8+ cytotoxic T lymphocytes enhances HIV-1 specific cytotoxic responses.

miRNAs dictate relevant virus-host interactions, offering new avenues for interventions to achieve an HIV remission. We aimed to enhance HIV-specific cytotoxic responses-a hallmark of natural HIV control- by miRNA modulation in T cells. We recruited 12 participants six elite controllers and six patients with chronic HIV infection on long-term antiretroviral therapy ("progressors"). Elite controllers exhibited stronger HIV-specific cytotoxic responses than the progressors, and their CD8+T cells showed a miRNA (hsa-miR-10a-5p) significantly downregulated. When we transfected ex vivo CD8+ T cells from progressors with a synthetic miR-10a-5p inhibitor, miR-10a-5p levels decreased in 4 out of 6 progressors, correlating with an increase in HIV-specific cytotoxic responses. The effects of miR-10a-5p inhibition on HIV-specific CTL responses were modest, short-lived, and occurred before day seven after modulation. IL-4 and TNF-α levels strongly correlated with HIV-specific cytotoxic capacity. Thus, inhibition of miR-10a-5p enhanced HIV-specific CD8+ T cell capacity in progressors. Our pilot study proves the concept that miRNA modulation is a feasible strategy to combat HIV persistence by enhancing specific cytotoxic immune responses, which will inform new approaches for achieving an antiretroviral therapy-free HIV remission.

Discovery and intranasal administration of a SARS-CoV-2 broadly acting neutralizing antibody with activity against multiple Omicron subvariants.

BACKGROUND: The continual emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants of concern, in particular the newly emerged Omicron (B.1.1.529) variant and its BA.X lineages, has rendered ineffective a number of previously FDA emergency use authorized SARS-CoV-2 neutralizing antibody therapies. Furthermore, those approved antibodies with neutralizing activity against Omicron BA.1 are reportedly ineffective against the subset of Omicron subvariants that contain a R346K substitution, BA.1.1, and the more recently emergent BA.2, demonstrating the continued need for discovery and characterization of candidate therapeutic antibodies with the breadth and potency of neutralizing activity required to treat newly diagnosed COVID-19 linked to recently emerged variants of concern. METHODS: Following a campaign of antibody discovery based on the vaccination of Harbor H2L2 mice with defined SARS-CoV-2 spike domains, we have characterized the activity of a large collection of spike-binding antibodies and identified a lead neutralizing human IgG1 LALA antibody, STI-9167. FINDINGS: STI-9167 has potent, broad-spectrum neutralizing activity against the current SARS-COV-2 variants of concern and retained activity against each of the tested Omicron subvariants in both pseudotype and live virus neutralization assays. Furthermore, STI-9167 nAb administered intranasally or intravenously provided protection against weight loss and reduced virus lung titers to levels below the limit of quantitation in Omicron-infected K18-hACE2 transgenic mice. CONCLUSIONS: With this established activity profile, a cGMP cell line has been developed and used to produce cGMP drug product intended for intravenous or intranasal use in human clinical trials. FUNDING: Funded by CRIPT (no. 75N93021R00014), DARPA (HR0011-19-2-0020), and NCI Seronet (U54CA260560).

Assessing costs of a hypertension program in primary care: evidence from the HEARTS program in Mexico / Evaluación de los costos de un programa de hipertensión en la atención primaria: datos del programa HEARTS en México / Avaliação dos custos de um programa de hipertensão na atenção primária: evidências do programa HEARTS no México

ABSTRACT Objective. In 2021, Mexico launched the HEARTS program to improve the prevention and control of cardiovascular disease (CVD) risk factors in 20 primary care facilities in the states of Chiapas and Yucatán. This study projects the annual cost of program implementation and discusses budgetary implications for scaling up the program. Methods. We obtained district-level data on treatment protocols, medication costs, and other resources required to prevent and treat CVD. We used the HEARTS Costing Tool to estimate total and per-patient costs. A "partial implementation" scenario calculated the costs of implementing HEARTS if existing pharmacological treatment protocols are left in place. The second scenario, "full implementation," examined costs if programs use HEARTS pharmacological protocol. Results. Respectively in the partial and full implementation scenarios, total annual costs to implement and operate HEARTS were $260 023 ($32.1 per patient/year) and $255 046 ($31.5 per patient/year) in Chiapas, and $1 000 059 ($41.3 per patient/year) and $1 013 835 ($43.3 per patient/year) in Yucatán. In Chiapas, adopting HEARTS standardized treatment protocols resulted in a 9.7 % reduction in annual medication expenditures relative to maintaining status-quo treatment approaches. In Yucatán, adoption was $12 875 more expensive, in part because HEARTS hypertension treatment regimens were more intensive than status quo regimens. Conclusion. HEARTS in the Americas offers a standardized strategy to treating and controlling CVD risk factors. In Mexico, approaches that may lead to improved program affordability include adoption of the recommended HEARTS treatment protocols with preferred medications and task shifting of services from physicians to nurses and other providers.

RESUMEN Objetivo. En el año 2021, México puso en marcha el programa HEARTS para mejorar la prevención y el control de los factores de riesgo de las enfermedades cardiovasculares en 20 centros de atención primaria en los estados de Chiapas y Yucatán. En este estudio se estima el costo anual de la ejecución del programa y se abordan las implicaciones presupuestarias para su ampliación. Métodos. Se obtuvieron datos a nivel de distrito sobre los protocolos de tratamiento, los costos de los medicamentos y otros recursos necesarios para prevenir y tratar las enfermedades cardiovasculares. Se empleó la herramienta HEARTS para el cálculo de costos con el fin de estimar los costos totales y por paciente. En una situación de "implementación parcial", se calcularon los costos de ejecutar HEARTS si se mantienen los protocolos de tratamiento farmacológico existentes. En un segundo escenario de "implementación completa", se examinaron los costos de los programas que emplean el protocolo farmacológico de HEARTS. Resultados. En los escenarios de implementación parcial y total, respectivamente, los costos anuales totales para implementar y poner en marcha el paquete de medidas HEARTS fueron de US$ 260 023 (US$ 32,1 por paciente al año) y US$ 255 046 (US$ 31,5 por paciente al año) en Chiapas, y US$ 1 000 059 (US$ 41,3 por paciente al año) y US$ 1 013 835 (US$ 43,3 por paciente al año) en Yucatán. En Chiapas, la adopción de los protocolos de tratamiento estandarizados de HEARTS supuso una reducción de 9,7% en los gastos anuales de medicamentos en comparación con el mantenimiento de los enfoques de tratamiento ya establecidos. En Yucatán, la adopción fue US$ 12 875 más cara, en parte porque los esquemas de tratamiento para la hipertensión que se proponen en HEARTS fueron más intensivos que los esquemas ya establecidos. Conclusiones. El programa HEARTS en la Región de las Américas ofrece una estrategia estandarizada para tratar y controlar los factores de riesgo de las enfermedades cardiovasculares. En México, los enfoques que pueden conducir a una mayor asequibilidad del programa incluyen la adopción de los protocolos de tratamiento recomendados de HEARTS con medicamentos de preferencia y la distribución de tareas de los servicios para que pasen del personal médico al personal de enfermería y otros prestadores de atención de salud.

RESUMO Objetivo. Em 2021, o México lançou o programa HEARTS para melhorar a prevenção e o controle dos fatores de risco de doenças cardiovasculares (DCV) em 20 unidades básicas de saúde nos estados de Chiapas e Yucatán. Este estudo projeta o custo anual de implementação do programa e discute as implicações orçamentárias para sua expansão. Métodos. Foram obtidos dados de nível distrital sobre protocolos de tratamento, custos de medicamentos e outros recursos necessários para prevenir e tratar a DCV. A ferramenta de cálculo de custos do HEARTS foi usada para estimar os custos totais e por paciente. Um cenário de "implementação parcial" calculou os custos de implementação do HEARTS se os protocolos de farmacoterapia existentes forem mantidos em vigor. O segundo cenário, "implementação plena", examinou os custos se os programas utilizassem o protocolo de farmacoterapia do HEARTS. Resultados. Respectivamente nos cenários de implementação parcial e plena, os custos anuais totais para implementar e operar o HEARTS foram de US$ 260 023 (US$ 32,1 por paciente/ano) e US$ 255 046 (US$ 31,5 por paciente/ano) em Chiapas, e $1 000 059 (US$ 41,3 por paciente/ano) e US$ 1 013 835 (US$ 43,30 por paciente/ano) em Yucatán. Em Chiapas, a adoção de protocolos de tratamento padronizados do HEARTS resultou em uma redução de 9,7% nos gastos anuais com medicamentos em relação à manutenção das condutas atuais (status quo). Em Yucatán, a adoção foi US$ 12 875 mais cara, em parte porque os regimes de tratamento de hipertensão do HEARTS eram mais intensivos do que os regimes atuais. Conclusão. A HEARTS nas Américas oferece uma estratégia padronizada para tratar e controlar os fatores de risco de DCV. No México, abordagens que podem levar a uma melhor acessibilidade do programa incluem a adoção dos protocolos de tratamento recomendados do HEARTS com medicamentos preferidos e a realocação de tarefas de médicos para enfermeiros e outros profissionais.

Hepatitis C virus drugs that inhibit SARS-CoV-2 papain-like protease synergize with remdesivir to suppress viral replication in cell culture.

Effective control of COVID-19 requires antivirals directed against SARS-CoV-2. We assessed 10 hepatitis C virus (HCV) protease-inhibitor drugs as potential SARS-CoV-2 antivirals. There is a striking structural similarity of the substrate binding clefts of SARS-CoV-2 main protease (Mpro) and HCV NS3/4A protease. Virtual docking experiments show that these HCV drugs can potentially bind into the Mpro substrate-binding cleft. We show that seven HCV drugs inhibit both SARS-CoV-2 Mpro protease activity and SARS-CoV-2 virus replication in Vero and/or human cells. However, their Mpro inhibiting activities did not correlate with their antiviral activities. This conundrum is resolved by demonstrating that four HCV protease inhibitor drugs, simeprevir, vaniprevir, paritaprevir, and grazoprevir inhibit the SARS CoV-2 papain-like protease (PLpro). HCV drugs that inhibit PLpro synergize with the viral polymerase inhibitor remdesivir to inhibit virus replication, increasing remdesivir's antiviral activity as much as 10-fold, while those that only inhibit Mpro do not synergize with remdesivir.

Plitidepsin has potent preclinical efficacy against SARS-CoV-2 by targeting the host protein eEF1A.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) viral proteins interact with the eukaryotic translation machinery, and inhibitors of translation have potent antiviral effects. We found that the drug plitidepsin (aplidin), which has limited clinical approval, possesses antiviral activity (90% inhibitory concentration = 0.88 nM) that is more potent than remdesivir against SARS-CoV-2 in vitro by a factor of 27.5, with limited toxicity in cell culture. Through the use of a drug-resistant mutant, we show that the antiviral activity of plitidepsin against SARS-CoV-2 is mediated through inhibition of the known target eEF1A (eukaryotic translation elongation factor 1A). We demonstrate the in vivo efficacy of plitidepsin treatment in two mouse models of SARS-CoV-2 infection with a reduction of viral replication in the lungs by two orders of magnitude using prophylactic treatment. Our results indicate that plitidepsin is a promising therapeutic candidate for COVID-19.

Efectividad de la ozonopuntura en pacientes con artrosis cervical / Effectiveness of ozonepuncture in patients with cervical osteoarthritis

Introducción: La artrosis vertebral es frecuente. Las causas y procesos fisiopatológicos que contribuyen a su avance son variados. Se plantea que 80 % de las personas mayores de 55 años de edad presentan alteraciones en la columna cervical. Objetivo: Evaluar la efectividad de la ozonopuntura para el alivio de síntomas y signos en pacientes con artrosis cervical. Métodos: Se realizó un estudio de intervención terapéutica en 80 pacientes con artrosis cervical, atendidos en la Consulta de Ortopedia y remitidos al Servicio de Medicina Tradicional y Natural del Hospital General Docente Dr. Juan Bruno Zayas Alfonso de Santiago de Cuba, desde mayo del 2017 hasta marzo del 2019. Se dividieron aleatoriamente en 2 grupos: de estudio y de control, con 40 integrantes cada uno. Resultados: La enfermedad prevaleció en los pacientes de 50-59 años de edad, tanto del grupo de estudio como de control (35,0 y 30,0 %, respectivamente) y en el sexo femenino (85,0 % en el primero y 82,5 % en el segundo). La evolución fue buena y excelente en casi la totalidad de los integrantes del grupo de estudio. Conclusiones: La ozonopuntura fue efectiva en los pacientes con artrosis cervical y no se presentaron reacciones secundarias relacionadas con su aplicación. La evolución resultó satisfactoria y se observó una respuesta mayor en la desaparición de los síntomas y signos clínicos con la terapéutica acupuntural que con la convencional.

Introduction: The vertebral osteoarthritis is frequent. The pathophysiological causes and processes that contribute to its advance are varied. It is said that the 80 % of people over 55 years present cervical spine disorders. Objective: To evaluate the effectiveness of the ozonepuncture for the relief of symptoms and signs in patients with cervical osteoarthritis. Methods: A study of therapeutic intervention in 80 patients with cervical osteoarthritis was carried out, they were assisted in the Orthopedics Service and referred to the Traditional and Natural Medicine Service of Dr. Juan Bruno Zayas Alfonso Teaching General Hospital in Santiago de Cuba, from May, 2017 to March, 2019. They were divided at random in 2 groups: study and control groups, with 40 members each one. Results: The disease prevailed in the 50-59 age group, in both groups (35.0 and 30.0 %, respectively) and in the female sex (85.0 % in the first one and 82.5 % in the second). The clinical course was good and excellent in almost all the members of the study group. Conclusions: The ozonepuncture was effective in the patients with cervical osteoarthritis and adverse reactions related to its application were not presented. The clinical course was satisfactory and a higher response was observed in the disappearance of the symptoms and clinical signs with the acupuntural therapeutic than with the conventional one.

The Global Phosphorylation Landscape of SARS-CoV-2 Infection.

The causative agent of the coronavirus disease 2019 (COVID-19) pandemic, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has infected millions and killed hundreds of thousands of people worldwide, highlighting an urgent need to develop antiviral therapies. Here we present a quantitative mass spectrometry-based phosphoproteomics survey of SARS-CoV-2 infection in Vero E6 cells, revealing dramatic rewiring of phosphorylation on host and viral proteins. SARS-CoV-2 infection promoted casein kinase II (CK2) and p38 MAPK activation, production of diverse cytokines, and shutdown of mitotic kinases, resulting in cell cycle arrest. Infection also stimulated a marked induction of CK2-containing filopodial protrusions possessing budding viral particles. Eighty-seven drugs and compounds were identified by mapping global phosphorylation profiles to dysregulated kinases and pathways. We found pharmacologic inhibition of the p38, CK2, CDK, AXL, and PIKFYVE kinases to possess antiviral efficacy, representing potential COVID-19 therapies.

Efectividad de la analgesia quirúrgica acupuntural en pacientes operados de catarata / Effectiveness of the acupunctural surgical analgesia in patients operated for cataract

Introducción: La catarata constituye una de las principales causas de ceguera a escala mundial. Es más frecuente en los ancianos y el tratamiento de elección es el quirúrgico. Objetivo: Evaluar la efectividad de la analgesia acupuntural en pacientes operados de catarata. Métodos: Se realizó un estudio de intervención terapéutica en 136 pacientes mayores de 40 años con diagnóstico de catarata, operados en el Servicio de Oftalmología del Hospital General Docente Dr. Juan Bruno Zayas Alfonso de Santiago de Cuba en el período 2016-2018. La muestra se dividió aleatoriamente en 2 grupos (de estudio y de control), con 68 integrantes cada uno. Resultados: Predominaron los pacientes de 70 años y más y del sexo femenino. Las variables hemodinámicas no sufrieron modificaciones significativas con el proceder terapéutico. Conclusiones: La analgesia quirúrgica acupuntural fue más efectiva que la convencional, pues se logró una sedación mayor en casi la totalidad de los pacientes durante la operación, así como mayor permanencia de su efecto durante el período posoperatorio; asimismo, hubo menor número de afectados con edema y hemorragia.

Introduction: The cataract constitutes one of the main causes of blindness worldwide. It is more frequent in the elderly and the election treatment is the surgical one. Objective: To evaluate the effectiveness of the acupunctural analgesia in patients operated for cataract. Methods: A study of therapeutic intervention was carried out in 136 patients older than 40 years with diagnosis of cataract, operated in the Ophthalmology Service of Dr. Juan Bruno Zayas Alfonso Teaching General Hospital in Santiago de Cuba in the period 2016-2018. The sample was divided at random in 2 groups (study and control), with 68 members each one. Results: The 70 years and over patients of the female sex prevailed. The hemodynamic variables didn't suffer significant modifications with the therapeutical procedure. Conclusions: The acupunctural surgical analgesia was more effective than the conventional one, because a higher sedation was achieved in almost all the patients during the operation, as well as higher permanency of its effect during the postoperatory period; also, there was smaller number of patients affected with edema and hemorrhages.

Preoperative diagnosis of atypical pelvic leiomyoma and sarcoma: the potential role of diffusion-weighted imaging.

The objective of our study was to determine the utility of diffusion-weighted magnetic resonance (DWMR) to differentiate the atypical uterine leiomyomas and sarcomas, establishing a cut-off value of the apparent diffusion coefficient (ADC) to rule out the malignancy. We performed a diagnostic accuracy retrospective study including 10 patients with pelvic sarcomas and 17 patients with leiomyomas. Atypical morphological features in magnetic resonance (MR) studies occurred in 58.8% of the patients, leading to a significant number of indeterminate diagnoses. In contrast, ADC values were consistent for leiomyomas, sarcomas, primary tumours, recurrences, intrauterine and in the extrauterine pelvic locations. The ADC cut-off value was set in 1 (×10-3 mm2/s). Thus, the ADC values equal or superior to 1 × 10-3 mm2/s were always associated with a leiomyoma. The structural MR accuracy was 66.7%, reaching 100% when using DWMR with dichotomised ADC values. Diffusion-weighted imaging with the quantitative measurement of ADC may be considered a useful preoperative test for the differentiation of atypical leiomyomas from sarcomas. Impact statement What is already known on this subject? Papers reporting the utility of a diffusion-weighted MR for the diagnosis of uterine sarcomas are scarce and consist of a small series. However, the published results are consistent with our study, with the decreased ADCs in the case of malignancy. What do the results of this study add? The main differential characteristic of our study is that we selected only the atypical leiomyomas: they share sonographic and MR features with sarcomas, which often leads to an inaccurate diagnosis. This is also the first paper reporting on the role of DWMR with ADC for these types of tumours in extrauterine pelvic locations. We demonstrated a consistent relationship between dichotomised ADC values in leiomyomas/sarcomas for these particular cases and in recurrent tumours, with no overlap between both the groups, as a difference with the previous reports. What are the implications of these findings for clinical practice and/or further research? Our study can be considered as a proof of concept supporting DWMR with ADC measurement as a useful tool to enhance the diagnostic accuracy of MR, highlighting its value to rule out malignancy. Hence, DWMR seems to be a potential useful test to include in the preoperative evaluation of clinically atypical uterine tumours.

Internal Disequilibria and Phenotypic Diversification during Replication of Hepatitis C Virus in a Noncoevolving Cellular Environment.

Viral quasispecies evolution upon long-term virus replication in a noncoevolving cellular environment raises relevant general issues, such as the attainment of population equilibrium, compliance with the molecular-clock hypothesis, or stability of the phenotypic profile. Here, we evaluate the adaptation, mutant spectrum dynamics, and phenotypic diversification of hepatitis C virus (HCV) in the course of 200 passages in human hepatoma cells in an experimental design that precluded coevolution of the cells with the virus. Adaptation to the cells was evidenced by increase in progeny production. The rate of accumulation of mutations in the genomic consensus sequence deviated slightly from linearity, and mutant spectrum analyses revealed a complex dynamic of mutational waves, which was sustained beyond passage 100. The virus underwent several phenotypic changes, some of which impacted the virus-host relationship, such as enhanced cell killing, a shift toward higher virion density, and increased shutoff of host cell protein synthesis. Fluctuations in progeny production and failure to reach population equilibrium at the genomic level suggest internal instabilities that anticipate an unpredictable HCV evolution in the complex liver environment.IMPORTANCE Long-term virus evolution in an unperturbed cellular environment can reveal features of virus evolution that cannot be explained by comparing natural viral isolates. In the present study, we investigate genetic and phenotypic changes that occur upon prolonged passage of hepatitis C virus (HCV) in human hepatoma cells in an experimental design in which host cell evolutionary change is prevented. Despite replication in a noncoevolving cellular environment, the virus exhibited internal population disequilibria that did not decline with increased adaptation to the host cells. The diversification of phenotypic traits suggests that disequilibria inherent to viral populations may provide a selective advantage to viruses that can be fully exploited in changing environments.

Favipiravir can evoke lethal mutagenesis and extinction of foot-and-mouth disease virus.

Antiviral agents are increasingly considered an option for veterinary medicine. An understanding of their mechanism of activity is important to plan their administration either as monotherapy or in combination with other agents. Previous studies have shown that the broad spectrum antiviral agent favipiravir (T-705) and its derivatives T-1105 and T-1106 are efficient inhibitors of foot-and-mouth disease virus (FMDV) replication in cell culture and in vivo. However, no mechanism for their activity against FMDV has been proposed. In the present study we show that favipiravir (T-705) can act as a lethal mutagen for FMDV in cell culture. Evidence includes virus extinction associated with increase in mutation frequency in the mutant spectrum of 860 residues of the 3D (polymerase)-coding region, and a decrease of specific infectivity while the consensus nucleotide sequence of the region analyzed remained invariant. The mutational spectrum evoked by favipiravir differs from that observed with other viruses in that no predominant transition type is observed, indicating that a movement towards A,U- or G,C-rich regions of sequence space is not a prerequisite for virus extinction. We discuss prospects for the use of favipiravir to assist in the control of FMDV, and its possible broader use in veterinary medicine as an extension of its current status as antiviral agent for human influenza virus.

Distance effects during polyprotein processing in the complementation between defective FMDV RNAs.

Passage of foot-and-mouth disease virus (FMDV) in BHK-21 cells resulted in the segmentation of the viral genome into two defective RNAs lacking part of either the L- or the capsid-coding region. The two RNAs are infectious by complementation. Electroporation of L-defective RNA in BHK-21 cells resulted in the accumulation of the precursor P3 located away from the deleted sequence. Expression of L in trans led to the processing of P3, indicating that there is a connection between L protease activity and the secondary cleavages carried out by 3C protease within P3. These results suggest that the complementation mechanism between defective RNAs is not restricted to supplying the L and capsid proteins but that distance effects on polyprotein processing events are also implicated.