Is gene therapy a good therapeutic approach for HIV-positive patients?

Despite advances and options available in gene therapy for HIV-1 infection, its application in the clinical setting has been challenging. Although published data from HIV-1 clinical trials show safety and proof of principle for gene therapy, positive clinical outcomes for infected patients have yet to be demonstrated. The cause for this slow progress may arise from the fact that HIV is a complex multi-organ system infection. There is uncertainty regarding the types of cells to target by gene therapy and there are issues regarding insufficient transduction of cells and long-term expression. This paper discusses state-of-the-art molecular approaches against HIV-1 and the application of these treatments in current and ongoing clinical trials

Significance of Premature Stop Codons in \u3cem\u3eenv\u3c/em\u3e of Simian Immunodeficiency Virus

The location of the translational termination codon for the transmembrane protein (TMP) varies in three infectious molecular clones of simian immunodeficiency virus from macaques (SIVmac). The SIVmac251 and SIVmac142 infectious clones have premature stop signals that differ in location by one codon; transfection of these DNAs into human HUT-78 cells yielded virus with a truncated TMP (28 to 30 kilodaltons [kDa]). The SIVmac239 infectious clone does not have a premature stop codon in its TMP-coding region. Transfection of HUT-78 cells with this clone initially yielded virus with a full-length TMP (41 kDa). At 20 to 30 days posttransfection, SIVmac239 virus with a 41-kDa TMP gradually disappeared coincident with the emergence of a virus with a 28-kDa TMP. Virus production dramatically increased in parallel with the emergence of a virus with a 28-kDa TMP. Sequence analysis of viral DNAs from these cultures showed that premature stop codons arising by point mutation were responsible for the change in size of the TMP with time. A similar selective pressure for truncated forms of TMP was observed when the SIVmac239 clone was transfected into human peripheral blood lymphocytes (PBL). In contrast, no such selective pressure was observed in macaque PBL. When the SIVmac239 clone was transfected into macaque PBL and the resultant virus was serially passaged in macaque PBL, the virus replicated very well and maintained a 41-kDa TMP for 80 days in culture. Macaque monkeys were infected with SIVmac239 having a 28-kDa TMP; virus subsequently recovered from T4-enriched lymphocytes of peripheral blood showed only the 41-kDa form of TMP. These results indicate that the natural form of TMP in SIVmac is the full-length 41-kDa TMP, just as in human immunodeficiency virus type 1. Viruses with truncated forms of TMP appear to result from mutation and selection during propagation in unnatural human cells

The United States of America and Scientific Research

To gauge the current commitment to scientific research in the United States of America (US), we compared federal research funding (FRF) with the US gross domestic product (GDP) and industry research spending during the past six decades. In order to address the recent globalization of scientific research, we also focused on four key indicators of research activities: research and development (R&D) funding, total science and engineering doctoral degrees, patents, and scientific publications. We compared these indicators across three major population and economic regions: the US, the European Union (EU) and the People's Republic of China (China) over the past decade. We discovered a number of interesting trends with direct relevance for science policy. The level of US FRF has varied between 0.2% and 0.6% of the GDP during the last six decades. Since the 1960s, the US FRF contribution has fallen from twice that of industrial research funding to roughly equal. Also, in the last two decades, the portion of the US government R&D spending devoted to research has increased. Although well below the US and the EU in overall funding, the current growth rate for R&D funding in China greatly exceeds that of both. Finally, the EU currently produces more science and engineering doctoral graduates and scientific publications than the US in absolute terms, but not per capita. This study's aim is to facilitate a serious discussion of key questions by the research community and federal policy makers. In particular, our results raise two questions with respect to: a) the increasing globalization of science: “What role is the US playing now, and what role will it play in the future of international science?”; and b) the ability to produce beneficial innovations for society: “How will the US continue to foster its strengths?

Retroviral Vector and Cell-Based Assay for Measuring the Mutation Rate of Retroviruses Employing Same

Lentiviral-based retrovirus vectors and an in vivo mutation rate assay employing them. More particularly, an assay for directly determining the in vivo mutation rate of HIV-1

Biological Safety: Principles and Practices

Biological safety and biosecurity protocols are essential to the reputation and responsibility of every scientific institution, whether research, academic, or production. Every risk―no matter how small―must be considered, assessed, and properly mitigated. If the science isn\u27t safe, it isn\u27t good. Now in its fifth edition, Biological safety: Principles and Practices remains the most comprehensive biosafety reference. Led by editors Karen Byers and Dawn Wooley, a team of expert contributors have outlined the technical nuts and bolts of biosafety and biosecurity within these pages. This book presents the guiding principles of laboratory safety, including: the identification, assessment, and control of the broad variety of risks encountered in the lab; the production facility; and, the classroom. Specifically, Biological Safety covers protection and control elements―from biosafety level cabinets and personal protection systems to strategies and decontamination methods administrative concerns in biorisk management, including regulations, guidelines, and compliance various aspects of risk assessment covering bacterial pathogens, viral agents, mycotic agents, protozoa and helminths, gene transfer vectors, zooonotic agents, allergens, toxins, and molecular agents as well as decontamination, aerobiology, occupational medicine, and training A resource for biosafety professionals, instructors, and those who work with pathogenic agents in any capacity, Biological safety is also a critical reference for laboratory managers, and those responsible for managing biohazards in a range of settings, including basic and agricultural research, clinical laboratories, the vivarium, field study, insectories, and greenhouses.https://corescholar.libraries.wright.edu/books/1248/thumbnail.jp

Duplex Quantitative Polymerase Chain Reaction Assay for Detection of Adenoviral and Lentiviral Vectors

The ability to detect and quantify viral vector sequences from a variety of clinical sample types is crucial to performing biosafety risk assessments. Viral vector-mediated gene transfer studies are often performed in animals, and these animals must be placed under appropriate biocontainment conditions to protect the workers and environment. Data on the shedding of viral vectors from animals are limited, and the sample types are challenging for polymerase chain reaction (PCR). For this reason, we developed a quantitative PCR assay that could be used for such purpose. We designed a sequence-specific, probe-binding method for detecting adenoviral and lentiviral vector sequences. A duplex strategy was used that included a quality control sequence to be amplified in the same reaction as the viral vector. This sequence provided an internal control for normalization of noncellular sample types, such as animal excretions that inherently lack a natural control sequence. The new assay was used to establish the efficiency of reverse transcription and to detect viral genomes in stocks of whole virus particles. We identified sets of primers and probes for both adenoviral and lentiviral sequences that work well together with no interference. The average conversion rate of RNA into complementary DNA was 18.5%. The new quantitative PCR assay was efficient and specific, and it measured successfully the number of viral genomes in stocks of whole virus particles. This assay could be used to detect adenoviral and lentiviral vector sequences for biosafety and other research purposes

Duplex Quantitative Polymerase Chain Reaction Assay for Detection of Adenoviral and Lentiviral Vectors

The ability to detect and quantify viral vector sequences from a variety of clinical sample types is crucial to performing biosafety risk assessments. Viral vector-mediated gene transfer studies are often performed in animals, and these animals must be placed under appropriate biocontainment conditions to protect the workers and environment. Data on the shedding of viral vectors from animals are limited, and the sample types are challenging for polymerase chain reaction (PCR). For this reason, we developed a quantitative PCR assay that could be used for such purpose. We designed a sequence-specific, probe-binding method for detecting adenoviral and lentiviral vector sequences. A duplex strategy was used that included a quality control sequence to be amplified in the same reaction as the viral vector. This sequence provided an internal control for normalization of noncellular sample types, such as animal excretions that inherently lack a natural control sequence. The new assay was used to establish the efficiency of reverse transcription and to detect viral genomes in stocks of whole virus particles. We identified sets of primers and probes for both adenoviral and lentiviral sequences that work well together with no interference. The average conversion rate of RNA into complementary DNA was 18.5%. The new quantitative PCR assay was efficient and specific, and it measured successfully the number of viral genomes in stocks of whole virus particles. This assay could be used to detect adenoviral and lentiviral vector sequences for biosafety and other research purposes

COVID 19 What You Need to Know

This is the first presentation in the new Shelter-in-Place lecture series. This first lecture is two-fold. First, it covers the basics of the COVID-19 virus including: how it spreads, possible origin, and other pertinent information. This portion of the presentation was followed by an overview of the international recruitment efforts in response to the virus, including: student perspectives, recruitment methods, and other important information