A universal fluorescence biosensor based on rolling circle amplification and locking probe for DNA detection.

A stable DNA signal amplification sensor was developed on account of rolling circle amplification (RCA). This sensor includes target DNA-controlled rolling circle amplification technology and locking probe DNA replacement technology, which can be used to detect DNA fragments with genetic information, thus constructing a biosensor for universal detection of DNA. This study takes the homologous DNA of human immunodeficiency virus (HIV) and let-7a as examples to describe this biosensor. The padlock probe is first cyclized by T4 DNA ligase in response to the target's reaction with it. Then, rolling cycle amplification is initiated by Phi29 DNA polymerase, resulting in the formation of a lengthy chain with several triggers. These triggers can open the locked probe LP1 with the fluorescence signal turned off, so that it can continue to react with H2 to form a stable H1-H2 double strand. This regulates the distance between B-DNA modified by the quenching group and H1 modified by fluorescent group, and the fluorescence signal is recovered.

Advanced Therapies for Human Immunodeficiency Virus.

Human Immunodeficiency Virus (HIV) remains a significant global health challenge with approximately 38 million people currently having the virus worldwide. Despite advances in treatment development, the virus persists in the human population and still leads to new infections. The virus has a powerful ability to mutate and hide from the human immune system in reservoirs of the body. Current standard treatment with antiretroviral therapy effectively controls viral replication but requires lifelong adherence and does not eradicate the virus. This review explores the potential of Advanced Therapy Medicinal Products as novel therapeutic approaches to HIV, including cell therapy, immunisation strategies and gene therapy. Cell therapy, particularly chimeric antigen receptor T cell therapy, shows promise in preclinical studies for targeting and eliminating HIV-infected cells. Immunisation therapies, such as broadly neutralising antibodies are being investigated to control viral replication and reduce reservoirs. Despite setbacks in recent trials, vaccines remain a promising avenue for HIV therapy development. Gene therapy using technologies like CRISPR/Cas9 aims to modify cells to resist HIV infection or eliminate infected cells. Challenges such as off-target effects, delivery efficiency and ethical considerations persist in gene therapy for HIV. Future directions require further research to assess the safety and efficacy of emerging therapies in clinical trials. Combined approaches may be necessary to achieve complete elimination of the HIV reservoir. Overall, advanced therapies offer new hope for advancing HIV treatment and moving closer to a cure.

Electrochemiluminescence of Ultrasmall Silica Nanoparticles from Size Modulation and Multipath Surface State Adjustment for Ultrasensitive HIV-DNA Fragment Detection.

Here, ultrasmall SiO2 nanoparticles (u-SiO2 NPs, <5 nm) with obvious electrochemiluminescence (ECL) phenomenon, which was absent for conventional silica nanoparticles (c-SiO2 NPs), were reported. In a finite ultrasmall volume, the u-SiO2 NPs exhibited increasing ground state energy and higher optical absorption strength due to the electron-hole confinement model and favored catalyzing the reaction through the rapid diffusion of bulk charge, resulting in apparent ECL emission. Then, Zn2+-induced u-SiO2 nanoaggregates (Zn/u-SiO2-Ov nAGG) were synthesized and exhibited improved ECL performance via multipath surface state adjustment of u-SiO2 from several aspects, including aggregation-induced ECL, the generation of oxygen vacancy (Ov), and more positive surface charge. In addition, an ECL biosensor was constructed for ultrasensitive human immunodeficiency virus-related deoxyribonucleic acid detection from 100 aM to 1 nM with a low limit of 50.48 aM, combining the ECL luminescence of Zn/u-SiO2-Ov nAGG with three-dimensional DNA nanomachine-mediated multioutput amplification for enhanced accuracy and sensitivity compared to the single-output method. Therefore, exploring the ECL of ultrasmall nanoparticles via the adjustment of size and surface state provided a valuable indication to a wider investigation and application of novel ECL materials for clinical diagnostic.

Roda Virtual discute acesso a medidas de prevenção ao HIV/Aids

A Plataforma IdeiaSUS Fiocruz realiza, nesta sexta-feira (21/6), das 14h às 16h, no canal da VideoSaúde Fiocruz no YouTube, a Roda Virtual Garantindo o direito de acesso universal à saúde: estratégias de disponibilização de PEP e PrEP. A temática surge do acompanhamento pela equipe da Curadoria em Saúde da IdeiaSUS da prática Planejamento Orientativo do Centro de Testagem e Aconselhamento (CTA/AS) sobre protocolo de PEP às farmácias do Vale do Ariano, desenvolvida em Juara (MT).

A green and simple method for enrichment of major diterpenoids from the buds of Wikstroemia chamaedaphne with macroporous resins and their activation of latent human immunodeficiency virus activity.

In this study, a green and efficient enrichment method for the four majors active diterpenoid components: pimelotide C, pimelotide A, simplexin, and 6α,7α-epoxy-5ß-hydroxy-12-deoxyphorbol-13-decanoate in the buds of Wikstroemia chamaedaphne was established using macroporous resin chromatography. The adsorption and desorption rates of seven macroporous resins were compared using static tests. The D101 macroporous resin exhibited the best performance. Static and dynamic adsorption tests were performed to determine the enrichment and purification of important bioactive diterpenoids in the buds of W. chamaedaphne. Diterpenoid extracts were obtained by using D101 macroporous resin from the crude extracts of W. chamaedaphne. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis demonstrated that most of the diterpenoids were enriched in diterpenoid extracts. These results confirmed that diterpenoids in the buds of W. chamaedaphne could be enriched using macroporous resin technology, and the enriched diterpenoid extracts showed more efficient activation of the latent human immunodeficiency virus. This study provides a novel strategy for discovering efficient and low-toxicity latency-reversing agents and a potential basis for the comprehensive development and clinical application of the buds of W. chamaedaphne.

Making a Monkey out of Human Immunodeficiency Virus/Simian Immunodeficiency Virus Pathogenesis: Immune Cell Depletion Experiments as a Tool to Understand the Immune Correlates of Protection and Pathogenicity in HIV Infection.

Understanding the underlying mechanisms of HIV pathogenesis is critical for designing successful HIV vaccines and cure strategies. However, achieving this goal is complicated by the virus's direct interactions with immune cells, the induction of persistent reservoirs in the immune system cells, and multiple strategies developed by the virus for immune evasion. Meanwhile, HIV and SIV infections induce a pandysfunction of the immune cell populations, making it difficult to untangle the various concurrent mechanisms of HIV pathogenesis. Over the years, one of the most successful approaches for dissecting the immune correlates of protection in HIV/SIV infection has been the in vivo depletion of various immune cell populations and assessment of the impact of these depletions on the outcome of infection in non-human primate models. Here, we present a detailed analysis of the strategies and results of manipulating SIV pathogenesis through in vivo depletions of key immune cells populations. Although each of these methods has its limitations, they have all contributed to our understanding of key pathogenic pathways in HIV/SIV infection.

Seroprevalence of human immunodeficiency virus in African blood donors: a systematic review and meta-analysis.

BACKGROUND: In developing countries, the safety of blood transfusions remains an important public health concern as it is associated with a higher risk of transfusion-transmissible infections (TTIs). In this study, we aimed to estimate the seroprevalence of HIV among blood donors in Africa and assess the temporal trends and regional differences within the continent through a systematic review and meta-analysis. METHODS: Seven electronic databases (PubMed, Web of Science, Cochrane, Scopus, HINARI, Global Index Medicus and Clinical. TRIAL: gov) were searched for relevant studies for our research. We included all primary studies that estimated the seroprevalence of HIV among blood donors in Africa with an age population from 16 to 65 years old, without language restrictions, from inception up to March 1st 2024. The pooled seroprevalence was estimated through the DerSimonian-Laird random effects model. The temporal trends and regional differences were assessed through subgroup and meta-regression analysis. FINDINGS: We obtained 122 studies that met our inclusion criteria, comprising 7,814,996 blood donors tested for HIV. Sixty-six percent of the studies were from Western and Eastern Africa. The pooled seroprevalence of HIV among blood donors in Africa was 2.66% (95% CI: 2.17-3.20%; I2 = 99.80%, p < 0.01). The highest prevalence was observed in the Central African region, 3.28% (95% CI: 2.57%-4.06%), followed by the Eastern 3.21% (95% CI: 2.12%-4.52%), and the Western 2.66% (95% CI: 1.93%-3.49%) regions. Lower prevalences were observed in the Northern region, 0.57% (95% CI: 0.0%-2.10%), followed by the Southern African region with 0.45% (95% CI: 0.16%-0.86%). We observed a temporal decreased trend of HIV prevalence. INTERPRETATION: The prevalence of HIV infection among African blood donors remains high and is not homogeneous across the continent. Efficient measures to strengthen HIV testing and prevent HIV transmission through blood transfusion are needed in Africa. Systematic review protocol registration: PROSPERO CRD42023395616. FUNDING: This article was supported by National Funds through FCT - Fundação para a Ciência e a Tecnologia,I.P., within CINTESIS, R&D Unit (reference UIDP/4255/2020).

From dysbiosis to defense: harnessing the gut microbiome in HIV/SIV therapy.

BACKGROUND: Although the microbiota has been extensively associated with HIV pathogenesis, the majority of studies, particularly those using omics techniques, are largely correlative and serve primarily as a basis for hypothesis generation. Furthermore, most have focused on characterizing the taxonomic composition of the bacterial component, often overlooking other levels of the microbiome. The intricate mechanisms by which the microbiota influences immune responses to HIV are still poorly understood. Interventional studies on gut microbiota provide a powerful tool to test the hypothesis of whether we can harness the microbiota to improve health outcomes in people with HIV. RESULTS: Here, we review the multifaceted role of the gut microbiome in HIV/SIV disease progression and its potential as a therapeutic target. We explore the complex interplay between gut microbial dysbiosis and systemic inflammation, highlighting the potential for microbiome-based therapeutics to open new avenues in HIV management. These include exploring the efficacy of probiotics, prebiotics, fecal microbiota transplantation, and targeted dietary modifications. We also address the challenges inherent in this research area, such as the difficulty in inducing long-lasting microbiome alterations and the complexities of study designs, including variations in probiotic strains, donor selection for FMT, antibiotic conditioning regimens, and the hurdles in translating findings into clinical practice. Finally, we speculate on future directions for this rapidly evolving field, emphasizing the need for a more granular understanding of microbiome-immune interactions, the development of personalized microbiome-based therapies, and the application of novel technologies to identify potential therapeutic agents. CONCLUSIONS: Our review underscores the importance of the gut microbiome in HIV/SIV disease and its potential as a target for innovative therapeutic strategies.

Padrão temporal, distribuição espacial e fatores associados a incidência de HIV/AIDS entre jovens no Brasil / Temporal pattern, spatial distribution, and factors associated with the incidence of HIV/AIDS among young people in Brazil / Patrón temporal, distribución espacial y factores asociados a la incidencia de la infección por el VIH/sida en jóvenes en Brasil

[RESUMO]. Objetivo. Descrever o padrão temporal e espacial e identificar os fatores associados a incidência de HIV/ AIDS entre jovens no Brasil. Método. Estudo ecológico que incluiu jovens brasileiros de 15 a 24 anos notificados com HIV/AIDS de 2001 a 2021. Utilizou-se o método joinpoint para a análise temporal. Aglomerados espaciais foram detectados pelos métodos Bayesiano, autocorrelação espacial, Getis-Ord Gi* e Varredura Scan. Quatro modelos de regressão não espacial e espacial foram usados para identificar fatores associados ao desfecho. Todas as análises estatísticas consideraram p < 0,05. Resultados. No Brasil, a incidência média foi de 12,29 por 100 000 habitantes, com aumento de 7,3% ao ano no período 2007-2014 e posterior decréscimo de 3,4% em 2014-2021. Observou-se padrão Alto/Alto e hots- pots, principalmente em municípios do Sul, Sudeste, Centro-Oeste e Norte. O cluster primário localizou-se em 572 municípios do Rio Grande do Sul e Santa Catarina e os maiores riscos relativos em Manaus (Amazonas) e Rondonópolis (Mato Grosso). A taxa de analfabetismo (β = -0,08), Índice de GINI (β = -3,74) e Cobertura da Estratégia de Saúde da Família (β = -0,70) apresentaram relação negativa com o desfecho. Em contrapar- tida, o Índice Firjan de Desenvolvimento Municipal (β = 2,37), o Índice de Vulnerabilidade Social (β = 6,30), o percentual de pessoas que recebem o Bolsa Família (β = 0,04) e renda per capita (β = 0,008) apresentaram associação positiva. Conclusão. Houve tendência de aumento da incidência de HIV/AIDS até 2014 com posterior declínio até 2021. Aglomerados de altas taxas concentraram-se, especialmente, em municípios das regiões Norte, Sul, Sudeste e Centro-Oeste. Indicadores de vulnerabilidade socioeconômica influenciam o desfecho positiva- mente ou negativamente, dependendo do território investigado.

[ABSTRACT]. Objective. To describe temporal and spatial patterns and identify the factors associated with the incidence of HIV/AIDS among young people in Brazil. Method. Ecological study of young Brazilians aged 15-24 years with reported HIV/AIDS, from 2001 to 2021. The Joinpoint method was used for the temporal analysis. Spatial clusters were detected using Bayesian methods, spatial autocorrelation, Getis-Ord Gi*, and scan techniques. Four non-spatial and spatial regression models were used to identify factors associated with the result. All statistical analyses considered p < 0.05. Results. In Brazil, the average incidence was 12.29 per 100 000 inhabitants, with an annual increase of 7.3% in the period 2007-2014 and a subsequent 3.4% decrease in 2014-2021. A high-high pattern and hotspots were observed, mainly in municipalities in the South, Southeast, Central-West, and North regions. The primary cluster was located in 572 municipalities in Rio Grande do Sul and Santa Catarina, with the highest relative risks in Manaus (Amazonas) and Rondonópolis (Mato Grosso). The illiteracy rate (β = -0.08), GINI Index (β = -3.74) and Family Health Strategy coverage (β = -0.70) were negatively associated with the result. In contrast, the Firjan Municipal Development Index (β = 2.37), Social Vulnerability Index (β = 6.30), percentage of Bolsa Família recipients (β = 0.04), and per capita income (β = 0.008) showed a positive association. Conclusion. There was an upward trend in the incidence of HIV/AIDS until 2014, followed by a decline until 2021. High-rate clusters were concentrated in municipalities in the North, South, Southeast and Central-West regions in particular. Indicators of socioeconomic vulnerability had positive or negative effects on the result, depending on the territory investigated.

[RESUMEN]. Objetivo. Describir el patrón temporal y espacial, y determinar los factores asociados a la incidencia de infec- ción por el VIH/sida en jóvenes en Brasil. Método. Estudio ecológico en jóvenes brasileños de 15 a 24 años con diagnóstico de infección por el VIH/ sida en el período 2001-2021. Para el análisis temporal se utilizó el método de regresión de puntos de infle- xión (joinpoint). Los conglomerados espaciales se detectaron con métodos Bayesianos y de autocorrelación espacial, Gi* de Getis-Ord y escaneo. Se utilizaron cuatro modelos de regresión espacial y no espacial para detectar los factores asociados al resultado. En todos los análisis estadísticos se estableció un valor de p < 0,05 como umbral de significación. Resultados. En Brasil, la incidencia media fue de 12,29 por 100 000 habitantes, con un aumento del 7,3% anual en el período 2007-2014 y una reducción posterior del 3,4% en el período 2014-2021. Se observó un patrón alto/alto y la presencia de puntos calientes, principalmente en municipios del Sur, Sudeste, Centro- Oeste y Norte. El principal conglomerado se localizó en 572 municipios de Rio Grande do Sul y Santa Catarina, y los riesgos relativos más altos se observaron en Manaus (Amazonas) y Rondonópolis (Mato Grosso). La tasa de analfabetismo (β = -0,08), el índice de Gini (β = -3,74) y la cobertura de la estrategia de salud familiar (β = -0,70) mostraron una asociación negativa con el resultado. En cambio, el índice de Firjan de desarrollo muni- cipal (β = 2,37), el índice de vulnerabilidad social (β = 6,30), el porcentaje de personas que reciben ayuda del programa de bienestar social Bolsa Família (β = 0,04) y los ingresos per cápita (β = 0,008) mostraron una asociación positiva. Conclusión. Hubo una tendencia al aumento de la incidencia de infección por el VIH/sida hasta el 2014, con una reducción posterior hasta el 2021. Los conglomerados de tasas elevadas se concentraron especialmente en los municipios de las regiones Norte, Sur, Sudeste y Centro-Oeste. Los indicadores de vulnerabilidad socioeconómica tienen una influencia positiva o negativa en el resultado, según el territorio investigado.

Manual operacional de tuberculose da OMS. Módulo 3: Diagnóstico -Testes para detecção de tuberculose infecção

O "Manual operacional da OMS sobre tuberculose. Módulo 3: Diagnóstico - Testes para detecção de tuberculose infecção" é um novo manual operacional sobre testes para a infecção por TB. Três classes de testes são agora recomendadas nas últimas diretrizes consolidadas sobre testes para infecção por tuberculose. Isso inclui pela primeira vez uma nova classe de testes cutâneos baseados em antígenos do Mycobacterium tuberculosis (TBSTs), e as duas classes existentes de testes: o teste cutâneo de tuberculina (TST) e os ensaios de liberação de interferon-gama (IGRAs). Os IGRAs e TBSTs utilizam antígenos específicos do complexo Mycobacterium tuberculosis e representam um avanço significativo em relação ao TST, que tem sido utilizado há mais de meio século. O manual operacional fornece como parceiros, pessoal de laboratório, clínicos, bem como ministérios da saúde e técnicos, com orientações detalhadas sobre como implementar as recomendações baseadas em evidências da OMS sobre testes de infecção por TB. O documento descreve os testes recomendados pela OMS, os procedimentos de teste, um algoritmo modelo e os passos necessários para ampliar os testes de infecção por TB dentro de um programa de saúde.

Coreactant-Free Zirconium Metal-Organic Framework with Dual Emission for Ratiometric Electrochemiluminescence Detection of HIV DNA.

Owing to the limitations of dual-signal luminescent materials and coreactants, constructing a ratiometric electrochemiluminescence (ECL) biosensor based on a single luminophore is a huge challenge. This work developed an excellent zirconium metal-organic framework (MOF) Zr-TBAPY as a single ECL luminophore, which simultaneously exhibited cathodic and anodic ECL without any additional coreactants. First, Zr-TBAPY was successfully prepared by a solvothermal method with 1,3,6,8-tetra(4-carboxyphenyl)pyrene (TBAPY) as the organic ligand and Zr4+ cluster as the metal node. The exploration of ECL mechanisms confirmed that the cathodic ECL of Zr-TBAPY originated from the pathway of reactive oxygen species (ROS) as the cathodic coreactant, which is generated by dissolved oxygen (O2), while the anodic ECL stemmed from the pathway of generated Zr-TBAPY radical itself as the anodic coreactant. Besides, N,N-diethylethylenediamine (DEDA) was developed as a regulator to ECL signals, which quenched the cathodic ECL and enhanced the anodic ECL, and the specific mechanisms of its dual action were also investigated. DEDA can act as the anodic coreactant while consuming the cathodic coreactant ROS. Therefore, the coreactant-free ratiometric ECL biosensor was skillfully constructed by combining the regulatory role of DEDA with the signal amplification reaction of catalytic hairpin assembly (CHA). The ECL biosensor realized the ultrasensitive ratio detection of HIV DNA. The linear range was 1 fM to 100 pM, and the limit of detection (LOD) was as low as 550 aM. The outstanding characteristic of Zr-TBAPY provided new thoughts for the development of ECL materials and developed a new way of fabricating the coreactant-free and single-luminophore ratiometric ECL platform.

Integração da vigilância e atenção à saúde no tratamento da sífilis gestacional: análise dos indicadores do PQAVS e do Previne Brasil na Paraíba / Integration of surveillance and health care in the management of gestational syphilis: analysis of PQAVS and Previne Brasil indicators in Paraíba / Integración de la vigilancia y la atención a la salud en el tratamiento de la sífilis gestacional: análisis de los indicadores PQAVS y Previne Brasil en Paraíba

Este artigo se baseia em um estudo feito com o objetivo de analisar indicadores sobre a testagem da sífilis na gestação no Programa de Qualificação das Ações de Vigilância em Saúde (PQAVS) e no Programa Previne Brasil no estado da Paraíba, e também de levantar aspectos do tratamento terapêutico para sífilis gestacional. Trata-se de uma pesquisa descritiva-exploratória, na qual foram sistematizados dados do indicador 11, testes por gestantes, do PQAVS e do indicador de desempenho da Atenção Primária à Saúde (APS), com base na proporção de gestantes que realizaram exames de sífilis e HIV durante o pré-natal em 2020; também foi feita a sistematização do webquestionário direcionado a profissionais da APS (médicos/enfermeiros) e autoaplicado sobre a atuação e tratamento terapêutico para sífilis gestacional. Dos 223 municípios da Paraíba, apenas 12% atingiram a meta do PQAVS e 39% a do Previne Brasil em 2020. Em relação ao webquestionário, houve a participação de 142 profissionais, dos quais 85% realizam o tratamento terapêutico preconizado pelo Ministério da Saúde para a APS. Desse modo, deve ser ressaltada a importância da ampliação da oferta de testes para sífilis, dos insumos para o tratamento adequado e da qualificação dos profissionais e da informação em saúde.

This article is based on a study to analyze indicators on syphilis testing during pregnancy in the PQAVS - Programa de Qualificação das Ações de Vigilância em Saúde (Health Surveillance Actions Qualification Programme) and in the Programa Previne Brasil (Previne Brasil Programme) in the state of Paraíba, Brazil, and also to survey aspects of the therapeutic management for gestational syphilis. It is a descriptive-exploratory research, in which data from indicator 11, tests for pregnant women, from the PQAVS and from the Primary Health Care (PHC) performance indicator, based on the proportion of pregnant women with syphilis and HIV tests during prenatal care in 2020 were systematised; in addition to this systematization, a self-administered webquestionnaire on the performance and therapeutic management for gestational syphilis by professionals (doctors/nurses) from the PHC was also systematised. Taking into account the 223 municipalities in Paraíba, only 12% reached the PQAVS goal and 39% reached the Previne Brasil goal in 2020. Regarding the webquestionnaire, 85% of the 142 professionals who answered it, carry out the therapeutic management recommended by the Ministry of Health for the PHC. Thus, it is fundamental to emphasise the importance of expanding the supply of tests for syphilis, supplies for adequate treatment, and the qualification of health professionals and information.

El presente artículo se basa en un estudio efectuado con el objetivo de analizar indicadores sobre la prueba de sífilis durante el embarazo en el PQAVS - Programa de Qualificação das Ações de Vigilância em Saúde (Programa de Calificación para Acciones de Vigilancia en Salud) y en el Programa Previne Brasil en el estado de Paraíba, Brasil, y de resaltar aspectos del tratamiento terapéutico de la sífilis gestacional. Se trata de una investigación descriptiva-exploratoria, en la que se sistematizaron datos del indicador 11, pruebas realizadas por embarazadas, del PQAVS y del indicador de desempeño de la Atención Primaria de Salud (APS), a partir de la proporción de gestantes que se sometieron a pruebas de sífilis y de HIV durante la atención prenatal en 2020; también se sistematizóel cuestionario web dirigido a profesionales de la APS (médicos/enfermeros) y autoadministrado sobre el desempeño y el tratamiento terapéutico de la sífilis gestacional. De los 223 municipios de Paraíba, apenas 12% alcanzaron la meta del PQAVS y 39% lograron la meta del Previne Brasil en 2020. En relación al cuestionario web, participaron 142 profesionales, de los cuales 85% realizan el tratamiento terapéutico recomendado por el Ministerio de Salud para la APS. Así, es fundamental la importancia de ampliar la oferta de pruebas para la sífilis, de los medicamentos para el tratamiento adecuado, la calificación de los profesionales e la información relacionada a la salud.

Comparative study of the interactions between C60 fullerene and SARS-CoV-2, HIV, eukaryotic, and bacterial model membranes: Insights into antimicrobial strategies with C60-peptide hybrids.

The neutrophil-derived peptide, indolicidin, and the sphere-shaped carbon nanoparticle, C60, are contemporary components capable of acting as bactericides and virucides, among others. Herein, the coarse-grained molecular dynamics simulation method was used to simulate the interactions of gram-negative bacteria, eukaryotes, human immunodeficiency virus (HIV), and SARS-COV-2 membrane models with indolicidin, C60s, and C60-indolicidin hybrids. Our results demonstrated that the carbon nanoparticle penetrated all membrane models, except the bacterial membrane, which remained impenetrable to both the peptide and C60. Additionally, the membrane thickness did not change significantly. The peptide floated above the membranes, with only the side chains of the tryptophan (Trp)-rich site slightly permeating the membranes. After achieving stable contact between the membrane models and nanoparticles, the infiltrated C60s interacted with the unsaturated tail of phospholipids. The density results showed that C60s stayed close to indolicidin and continued to interact with it even after penetration. Indolicidin, especially its Trp-rich site, exhibited more contact with the head and tail of neutral phospholipids compared to other phospholipids. Moreover, both particles interacted with different kinds of glycosphingolipids located in the eukaryote membrane. This investigation has the potential to advance our knowledge of novel approaches to combat antimicrobial resistance.

Direct Analysis of HIV mRNA m6A Methylation by Nanopore Sequencing.

The post-transcriptional processing and chemical modification of HIV RNA are understudied aspects of HIV virology, primarily due to the limited ability to accurately map and quantify RNA modifications. Modification-specific antibodies or modification-sensitive endonucleases coupled with short-read RNA sequencing technologies have allowed for low-resolution or limited mapping of important regulatory modifications of HIV RNA such as N6-methyladenosine (m6A). However, a high-resolution map of where these sites occur on HIV transcripts is needed for detailed mechanistic understanding. This has recently become possible with new sequencing technologies. Here, we describe the direct RNA sequencing of HIV transcripts using an Oxford Nanopore Technologies sequencer and the use of this technique to map m6A at near single nucleotide resolution. This technology also provides the ability to identify splice variants with long RNA reads and thus, can provide high-resolution RNA modification maps that distinguish between overlapping splice variants. The protocols outlined here for m6A also provide a powerful paradigm for studying any other RNA modifications that can be detected on the nanopore platform.

Bystander Effects and Profibrotic Interactions in Hepatic Stellate Cells during HIV and HCV Coinfection.

This study aims to explore the influence of coinfection with HCV and HIV on hepatic fibrosis. A coculture system was set up to actively replicate both viruses, incorporating CD4 T lymphocytes (Jurkat), hepatic stellate cells (LX-2), and hepatocytes (Huh7.5). LX-2 cells' susceptibility to HIV infection was assessed through measurements of HIV receptor expression, exposure to cell-free virus, and cell-to-cell contact with HIV-infected Jurkat cells. The study evaluated profibrotic parameters, including programed cell death, ROS imbalance, cytokines (IL-6, TGF-ß, and TNF-α), and extracellular matrix components (collagen, α-SMA, and MMP-9). The impact of HCV infection on LX-2/HIV-Jurkat was examined using soluble factors released from HCV-infected hepatocytes. Despite LX-2 cells being nonsusceptible to direct HIV infection, bystander effects were observed, leading to increased oxidative stress and dysregulated profibrotic cytokine release. Coculture with HIV-infected Jurkat cells intensified hepatic fibrosis, redox imbalance, expression of profibrotic cytokines, and extracellular matrix production. Conversely, HCV-infected Huh7.5 cells exhibited elevated profibrotic gene transcriptions but without measurable effects on the LX-2/HIV-Jurkat coculture. This study highlights how HIV-infected lymphocytes worsen hepatic fibrosis during HCV/HIV coinfection. They increase oxidative stress, profibrotic cytokine levels, and extracellular matrix production in hepatic stellate cells through direct contact and soluble factors. These insights offer valuable potential therapies for coinfected individuals.

Palm-Sized Lab-In-A-Magnetofluidic Tube Platform for Rapid and Sensitive Virus Detection.

Simple, sensitive, and accurate molecular diagnostics are critical for preventing rapid spread of infection and initiating early treatment of diseases. However, current molecular detection methods typically rely on extensive nucleic acid sample preparation and expensive instrumentation. Here, a simple, fully integrated, lab-in-a-magnetofluidic tube (LIAMT) platform is presented for "sample-to-result" molecular detection of virus. By leveraging magnetofluidic transport of micro/nano magnetic beads, the LIAMT device integrates viral lysis, nucleic acid extraction, isothermal amplification, and CRISPR detection within a single engineered microcentrifuge tube. To enable point-of-care molecular diagnostics, a palm-sized processor is developed for magnetofluidic separation, nucleic acid amplification, and visual fluorescence detection. The LIAMT platform is applied to detect SARS-CoV-2 and HIV viruses, achieving a detection sensitivity of 73.4 and 63.9 copies µL-1, respectively. Its clinical utility is further demonstrated by detecting SARS-CoV-2 and HIV in clinical samples. This simple, affordable, and portable LIAMT platform holds promise for rapid and sensitive molecular diagnostics of infectious diseases at the point-of-care.

Pathways to prevention: insights on stemming HIV outbreaks in Larkana, Pakistan.

In the past two decades, Pakistan has faced multiple human immunodeficiency virus outbreaks, with Larkana appearing to be the hub of such outbreaks. While the previous Larkana outbreaks happened in high-risk populations, the alarming outbreak in 2019 occurred in a low-risk paediatric population, raising several concerning questions. Human immunodeficiency virus infections spilling into the general population is indicative of a steady increase in the number of cases, and the failure of control strategies to stem the concentrated epidemic from evolving. Although several causative factors have been identified from previous outbreaks, the one that occurred in 2019 may have been influenced by an additional, hitherto unexplored factor; child sexual abuse. The current narrative review was planned to summarise human immunodeficiency virus risk factors and causes identified in previous Larkana epidemics, to explore potential reasons for the outbreaks in children, and to discuss possible steps needed for stemming human immunodeficiency virus outbreaks in Pakistan.

A novel simulation-based analysis of a stochastic HIV model with the time delay using high order spectral collocation technique.

The economic impact of Human Immunodeficiency Virus (HIV) goes beyond individual levels and it has a significant influence on communities and nations worldwide. Studying the transmission patterns in HIV dynamics is crucial for understanding the tracking behavior and informing policymakers about the possible control of this viral infection. Various approaches have been adopted to explore how the virus interacts with the immune system. Models involving differential equations with delays have become prevalent across various scientific and technical domains over the past few decades. In this study, we present a novel mathematical model comprising a system of delay differential equations to describe the dynamics of intramural HIV infection. The model characterizes three distinct cell sub-populations and the HIV virus. By incorporating time delay between the viral entry into target cells and the subsequent production of new virions, our model provides a comprehensive understanding of the infection process. Our study focuses on investigating the stability of two crucial equilibrium states the infection-free and endemic equilibriums. To analyze the infection-free equilibrium, we utilize the LaSalle invariance principle. Further, we prove that if reproduction is less than unity, the disease free equilibrium is locally and globally asymptotically stable. To ensure numerical accuracy and preservation of essential properties from the continuous mathematical model, we use a spectral scheme having a higher-order accuracy. This scheme effectively captures the underlying dynamics and enables efficient numerical simulations.

Pulmonary symptoms associated with heavy alcohol consumption among people living with HIV: an analysis of the NHANES 1999-2010.

AIM: This matched case-control study aimed to provide epidemiologic evidence of increased burden of respiratory symptoms and pulmonary function decline among people living with human immunodeficiency virus (HIV) and a history of heavy alcohol consumption. METHODS: Cases were participants with HIV (PWH; n = 75, 33%), and controls were participants without HIV (PWoH; n = 150, 67%). PWH were matched to PWoH by age and sex in the ratio of 1:2. Eligible participants responded to the respiratory health National Health and Nutrition Examination Survey questionnaire [prolonged coughs (≥3 months), bringing up of phlegm (≥3 months), and a history of wheezing or whistling in the chest (past year)]. The effects of both alcohol and HIV on participants' pulmonary function were determined using linear regression analysis. RESULTS: History of heavy alcohol consumption was more prevalent among PWH (40%) compared to PWoH (27%). PWH who had a history of heavy alcohol consumption had a higher prevalence of coughing most days (45% vs. 4%, P = .0010), bringing up phlegm most days (31% vs. 0%, P = .0012), and wheezing or whistling in the chest (40% vs. 20%, P = .058) compared to participants who did not heavily consume alcohol. Furthermore, a history of heavy alcohol consumption was associated with decreased forced expiratory volume (ml) in 1 s/forced vital capacity among PWH (ß = - 0.098 95% C.I. -0.16, -0.04, P = .03) after adjusting for having smoked at least 100 cigarettes in life. CONCLUSION: A history of heavy alcohol use increased respiratory symptoms and suppressed pulmonary function among people living with HIV. This study provides epidemiological evidence of the respiratory symptom burden of people living with HIV who have a history of heavy alcohol consumption.