Use ofcis- andtrans-Acting Viral Regulatory Sequences to Improve Expression of Human Immunodeficiency Virus Vectors in Human Lymphocytes

AbstractWe compared the efficiency of human immunodeficiency virus (HIV-1) vectors that express a marker gene (chloramphenicol acetyltransferase, CAT) using different promoter elements. In one vector, CAT was expressed under the control of an internal murine leukemia virus (MuLV) long terminal repeat (LTR). In other vectors, CAT production was regulated by the HIV-1 LTR; these vectors also contained the HIV-1tatgene andpolsequences reported to exertcis-acting positive effects on reverse transcription or gene expression. Vectors employing the Tat-driven HIV-1 LTR exhibited up to 500-fold greater CAT expression in Jurkat lymphocytes or human peripheral blood mononuclear cells compared with vectors using the internal MuLV LTR element as a promoter. This difference was not due to improved packaging of the vector RNA into virions, but to an improved level of gene expression in the target cells. Target cell CAT expression was two- to threefold higher for the vector containing thepolsequences and was only slightly less than that seen for atrans-complementedenv-deleted provirus. These results indicate that defective HIV-1 vectors with efficiencies of gene transfer and expression comparable with that of HIV-1 itself are feasible

Medication overuse headache, addiction and personality pathology: a controlled study by SWAP-200

Background: Medication Overuse Headache (MOH) is a type of chronic headache, whose mechanisms are still unknown. Some empirical investigations examining the addiction-like behaviors and processes, as well as personality characteristics underlying MOH development, reached contrasting findings. This study aimed at detecting personality and its disorders (PDs) in MOH patients, with a specific attention to the features of addiction. Methods: Eighty-eight MOH patients have been compared with two clinical populations including 99 patients with Substance Use Disorder (SUD) and 91 with PDs using the Shedler-Westen Assessment Procedure-200 (SWAP-200). MANCOVAs were performed to evaluate personality differences among MOH, SUD and PD groups, controlling for age and gender. Results: MOH patients showed lower traits of the SWAP-200’s clusters A and B disorders than SUD and PD patients, whom presented more severe levels of personality impairment. No differences in the SWAP-200’s cluster C have been found, indicating common personality features in these populations. At levels of specific PDs, MOH patients presented higher obsessive and dysphoric traits, as well as better overall psychological functioning than SUD and PD patients. Conclusions: The study supported the presence of a specific pattern of personality in MOH patients including obsessive (perfectionist) and dysphoric characteristics, as well as good enough psychological resources. No similarities with drug addicted and personality-disordered patients were found. Practitioners’ careful understanding of the personality of MOH patients may be useful to provide more effective treatment strategies and patient-tailored intervention programs

Lentiviral Vectors as Tools for the Study and Treatment of Glioblastoma

Glioblastoma (GBM) has the worst prognosis among brain tumors, hence basic biology, preclinical, and clinical studies are necessary to design effective strategies to defeat this disease. Gene transfer vectors derived from the most-studied lentivirus-the Human Immunodeficiency Virus type 1-have wide application in dissecting GBM specific features to identify potential therapeutic targets. Last-generation lentiviruses (LV), highly improved in safety profile and gene transfer capacity, are also largely employed as delivery systems of therapeutic molecules to be employed in gene therapy (GT) approaches. LV were initially used in GT protocols aimed at the expression of suicide factors to induce GBM cell death. Subsequently, LV were adopted to either express small noncoding RNAs to affect different aspects of GBM biology or to overcome the resistance to both chemo- and radiotherapy that easily develop in this tumor after initial therapy. Newer frontiers include adoption of LV for engineering T cells to express chimeric antigen receptors recognizing specific GBM antigens, or for transducing specific cell types that, due to their biological properties, can function as carriers of therapeutic molecules to the cancer mass. Finally, LV allow the setting up of improved animal models crucial for the validation of GBM specific therapies

View and review on viral oncology research

To date, almost one and a half million cases of cancer are diagnosed every year in the US and nearly 560,000 Americans are expected to die of cancer in the current year, more than 1,500 people a day (data from the American Cancer Society at http://www.cancer.org/). According to the World Health Organization (WHO), roughly 20% of all cancers worldwide results from chronic infections; in particular, up to 15% of human cancers is characterized by a viral aetiology with higher incidence in Developing Countries. The link between viruses and cancer was one of the pivotal discoveries in cancer research during the past Century. Indeed, the infectious nature of specific tumors has important implications in terms of their prevention, diagnosis, and therapy. In the 21st Century, the research on viral oncology field continues to be vigorous, with new significant and original studies on viral oncogenesis and translational research from basic virology to treatment of cancer. This review will cover different viral oncology aspects, starting from the history of viral oncology and moving to the peculiar features of oncogenic RNA and DNA viruses, with a special focus on human pathogens

New Evidence of Ancient Mitochondrial DNA of the Southern Andes (Calchaquí Valleys, Northwest Argentina, 3600–1900 Years BPA)

Genetic studies on pre-Hispanic populations of the Southern Andes have been increasing steadily in the last decade. Nevertheless, ancient DNA characterization of Formative Period archaeological human remains is particularly scant, especially for Northwest Argentina. In order to enlarge current information on genetic characterization of the first agricultural communities of the Southern Calchaquí Valleys, we present and discuss the first mitochondrial aDNA information obtained on samples dated to ca. 3600-1900 years BP from the Cajón Valley, Catamarca Province. Reproducible mtDNA HVR-1 sequences were obtained in 7 individuals. Mitochondrial HVR-1 haplotypes were assigned to three of the four founding haplogroups D1 (57.1%), C1 (28.5%) and B2 (14.2%), with absence of A2. Our results show that the Cajón Valley sample -with predominance of D1 and C1- differ from that commonly observed in ancient and modern Andean populations, which usually show a high prevalence of haplogroup B2. The fact that the Cajón Valley and Pampa Grande (Salta Province, Argentina) share a prevalence of haplogroup D1 could provide additional evidence to support possible genetic affinities between the Valleys and the eastern sub-Andean region during the Formative Period in Northwest Argentina, expanding the archaeological evidence of contact between both populations. Future complete mitogenomes analysis will provide substantial information to formulate new hypotheses about the origins and phylogenetic relationships between the individuals of the Cajón Valley and other groups from the Andes, Gran Chaco and the Amazon

How to break free: HIV-1 escapes from innovative therapeutic approaches

With nearly 38 million of people worldwide living with HIV-1 and no definitive cure available after almost 40 years of research, AIDS is still a major global public health issue. Modern antiretroviral therapies can achieve viral replication suppression to undetectable levels, thus allowing an almost normal life to HIV-1–infected individuals. However, the virus cannot be fully eradicated. This may lead over time to the accumulation of mutations in the viral genome and, eventually, to the emergence of drug-resistant viruses, which may affect the efficacy of the therapy and the patient’s quality of life. To overcome some of the limitations of the standard antiretroviral therapy, innovative therapeutic approaches such as “shock and kill” and immunotherapies, as well as technologies based on RNA interference and CRISPR-Cas9 genome editing are under investigation. Nevertheless, the virus may find a way to break free even from these novel strategies. In this review, we focus on the mechanisms that enable HIV-1 escape from the most advanced therapies and discuss some of the challenges to prevent this issue

Laser Scanning Confocal Microscopy: Application in Manufacturing and Research of Corneal Stem Cells

Laser scanning confocal microscopes (LSCMs) are powerful devices used to acquire high definition optical images by choosing the required depth selectively. The presence of specific laser beams and features such as fluorescence recovery after photobleaching (FRAP), fluores\u2010 cence lifetime imaging microscopy (FLIM), and fluorescence resonance energy transfer (FRET) allow to: i. increase the quality of the image; ii. observe and analyze subcellular organelles; iii. track the localization of any given labeled molecule within the cell; iv. identify specific areas within a tissue/organ (Figure 1). In parallel, the development and manufacturing of fluorescent probes (=fluorophores) characterized by low toxicity profiles are allowing to perform the above mentioned studies using living cell cultures or tissues that are not fixed. Furthermore, fluorescent proteins such as the Green Fluorescent Protein (GFP) and its derivatives allow to detect how the biosynthetic machinery of the cell works or a transgene (driven by a plasmid or a genetically engineered virus) is expressed (Figure 2) or a chimeric protein interacts with other cellular components. The aim of this chapter is therefore to describe how LSCM functions and features have helped vision sciences and regenerative medicine applications in the field of ophthalmology. The next sections will analyze how LSCM-based analyses have helped to: 1. evaluate how the ocular surface is formed; 2. define the role of p63 as stem cell marker; 3. set up quality control assays required for clinical applications of limbal stem cells in patients with limbal stem cell deficiency (LSCD); 4. validate the use of impression citology as a diagnostic tool for LSCD; 5. study gene therapy-based potential ways to treat rare genetic disorders of the ocular surface

Ebola Virus Entry: From Molecular Characterization to Drug Discovery

Ebola Virus Disease (EVD) is one of the most lethal transmissible infections, characterized by a high fatality rate, and caused by a member of the Filoviridae family. The recent large outbreak of EVD in Western Africa (2013\u207b2016) highlighted the worldwide threat represented by the disease and its impact on global public health and the economy. The development of highly needed anti-Ebola virus antivirals has been so far hampered by the shortage of tools to study their life cycle in vitro, allowing to screen for potential active compounds outside a biosafety level-4 (BSL-4) containment. Importantly, the development of surrogate models to study Ebola virus entry in a BSL-2 setting, such as viral pseudotypes and Ebola virus-like particles, tremendously boosted both our knowledge of the viral life cycle and the identification of promising antiviral compounds interfering with viral entry. In this context, the combination of such surrogate systems with large-scale small molecule compounds and haploid genetic screenings, as well as rational drug design and drug repurposing approaches will prove priceless in our quest for the development of a treatment for EVD

Herpes simplex virus type 2 infection increases human immunodeficiency virus type 1 entry into human primary macrophages

Epidemiological and clinical data indicate that genital ulcer disease (GUD) pathogens are associated with an increased risk of human immunodeficiency virus type 1 (HIV-1) acquisition and/or transmission. Among them, genital herpes simplex virus type 2 (HSV-2) seems to play a relevant role. Indeed, the ability of HSV-2 to induce massive infiltration at the genital level of cells which are potential targets for HIV-1 infection may represent one of the mechanisms involved in this process. Here we show that infection of human primary macrophages (MDMs) by HSV-2 results in an increase of CCR5 expression levels on cell surface and allows higher efficiency of MDMs to support entry of R5 HIV-1 strains. This finding could strengthen, at the molecular level, the evidence linking HSV-2 infection to an increased susceptibility to HIV-1 acquisition

Un aporte antropogenético a la reconstrucción cultural de las comunidades afrobolivianas

En procura de su visibilización y el rescate de su cultura, el pueblo afroboliviano está buscando sus raíces originarias. El presente trabajo relata un estudio antropogenético realizado en las poblaciones afrodescendientes de Tocaña, Chijchipa, Mururata y San Joaquín, en Nor Yungas, Bolivia. La caracterización de las comunidades en base a marcadores genéticos y biodemografía ha permitido observar el alto grado de conservación del acervo africano. Se analiza la contribución del trabajo al proceso de búsqueda de identidad de la comunidad afroboliviana.In search of its visibility and the rescue of its culture, the AfroBolivian people are looking for their original roots. The present paper reports an anthropogenic study carried out in the Afro-descendant populations of Tocaña, Chijchipa, Mururata and San Joaquín, in Nor Yungas, Bolivia. The characterization of the communities based on genetic markers and biodemography has allowed to observe the high degree of conservation of the African heritage. We analyze the contribution of this work to the search process of identity of the afroboliviana community.Fil: Iudica, Celia Estela. Universidad de Buenos Aires. Facultad de Filosofía y Letras; Argentina. Universidad Nacional de Mar del Plata; ArgentinaFil: Parolin, María Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico; ArgentinaFil: Avena, Sergio Alejandro. Universidad de Buenos Aires. Facultad de Filosofía y Letras; Argentina. Universidad Maimónides; ArgentinaFil: Dejean, Cristina Beatriz. Universidad de Buenos Aires. Facultad de Filosofía y Letras; Argentina. Universidad Maimónides; ArgentinaFil: Carnese, Francisco Raul. Universidad de Buenos Aires. Facultad de Filosofía y Letras; Argentin