Expression and reactivation of HIV in a chemokine induced model of HIV latency in primary resting CD4+ T cells

<p>Abstract</p> <p>Background</p> <p>We recently described that HIV latent infection can be established <it>in vitro </it>following incubation of resting CD4+ T-cells with chemokines that bind to CCR7. The main aim of this study was to fully define the post-integration blocks to virus replication in this model of CCL19-induced HIV latency.</p> <p>Results</p> <p>High levels of integrated HIV DNA but low production of reverse transcriptase (RT) was found in CCL19-treated CD4+ T-cells infected with either wild type (WT) NL4.3 or single round envelope deleted NL4.3 pseudotyped virus (NL4.3- Δenv). Supernatants from CCL19-treated cells infected with either WT NL4.3 or NL4.3- Δenv did not induce luciferase expression in TZM-bl cells, and there was no expression of intracellular p24. Following infection of CCL19-treated CD4+ T-cells with NL4.3 with enhanced green fluorescent protein (EGFP) inserted into the <it>nef </it>open reading frame (NL4.3- Δnef-EGFP), there was no EGFP expression detected. These data are consistent with non-productive latent infection of CCL19-treated infected CD4+ T-cells. Treatment of cells with phytohemagluttinin (PHA)/IL-2 or CCL19, prior to infection with WT NL4.3, resulted in a mean fold change in unspliced (US) RNA at day 4 compared to day 0 of 21.2 and 1.1 respectively (p = 0.01; n = 5), and the mean expression of multiply spliced (MS) RNA was 56,000, and 5,000 copies/million cells respectively (p = 0.01; n = 5). In CCL19-treated infected CD4+ T-cells, MS-RNA was detected in the nucleus and not in the cytoplasm; in contrast to PHA/IL-2 activated infected cells where MS RNA was detected in both. Virus could be recovered from CCL19-treated infected CD4+ T-cells following mitogen stimulation (with PHA and phorbyl myristate acetate (PMA)) as well as TNFα, IL-7, prostratin and vorinostat.</p> <p>Conclusions</p> <p>In this model of CCL19-induced HIV latency, we demonstrate HIV integration without spontaneous production of infectious virus, detection of MS RNA in the nucleus only, and the induction of virus production with multiple activating stimuli. These data are consistent with <it>ex vivo </it>findings from latently infected CD4+ T-cells from patients on combination antiretroviral therapy, and therefore provide further support of this model as an excellent <it>in vitro </it>model of HIV latency.</p

Design of a modified MIMO antenna based on tweaked spherical fractal geometry for 5G New Radio (NR) band N258 (24.25–27.25 GHz) applications

This article describes a fractal-based MIMO antenna for 5G mm-wave mobile applications with micro-strip feeding. The proposed structure is a fractal-based spherical configuration that incorporates spherical slots of different iterations on the patch, as well as rectangular slots on the ground plane. These additions are meant to reduce patch isolation. The two-element MIMO antenna has closely spaced antenna elements that resonate at multiple frequencies, 9.5 GHz, 11.1 GHz, 13.4 GHz, 15.8 GHz, 21.1 GHz, and 26.6 GHz, in the frequency range of 8 to 28 GHz. The antenna’s broadest operational frequency range spans from 17.7 GHz to 28 GHz, encompassing a bandwidth of 10,300 MHz. Consequently, it is well-suited for utilization within the millimeter wave (mm wave) application, specifically for the 5G new radio frequency band n258, and partially covers some other bands X (8.9–9.9 GHz, 10.4–11.4 GHz), and Ku (13.1–13.7 GHz, 15.4–16.2 GHz). All the resonating bands have isolation levels below the acceptable range of (|S12| > −16 dB). The proposed antenna utilizes a FR4 material with dimension of 28.22 mm × 44 mm. An investigation is conducted to analyze the effectiveness of parameters of the antenna, including radiation pattern, surface current distributions and S parameters. Furthermore, an examination and assessment are conducted on the efficacy of the diversity system inside the multiple input multiple output (MIMO) framework. This evaluation encompasses the analysis of key performance metrics such as the envelope correlation coefficient (ECC), diversity gain (DG), and mean effective gain (MEG). All antenna characteristics are determined to be within a suitable range for this suggested MIMO arrangement. The antenna design underwent experimental validation and the simulated outcomes were subsequently verified

Self‐Medication Patterns Among Medical Students in South India

BackgroundSelf-medication results in wastage of resources, increases resistance of pathogens and generally causes serious health hazards such as adverse drug reactions, prolonged suffering and drug dependence. This study was undertaken to determine the reasons for self-medication and the pattern of self-medication among medical students. Method This cross-sectional descriptive study was conducted at the K.S. Hegde Medical Academy, Mangalore. The participants were medical students from first to final year. Medical students were selected through convenience sampling. The data was collected using a pre-tested semi-structured questionnaire. The data was analysed using SPSS version 16 and the results expressed as proportions.Results A total of 200 students, 121 (60.5%) female and 79 (39.5%) male, were included in the study. Of the medical students surveyed, self-medication was reported among 92%. The respondents who used self-medication found it to be time-saving in providing relief from minor ailments. The most common ailments for which self-medication were used were: the common cold (69%), fever (63%) and headache (60%). The students consulted their textbooks (39%) and seniors or classmates (38%) for the medications. Antipyretics (71%), analgesics (65%), antihistamines (37%) and antibiotics (34%) were the most common self-medicated drugs. Of the respondents, 33% were unaware of the adverse effects of the medication and 5% had experienced adverse reactions. The majority (64%) of students advised medications to others, more often to family and friends. Conclusion The prevalence of self-medication among medical students is high, facilitated by the easy availability of drugs and information from textbooks or seniors. A significant number of students are unaware of the adverse effects of the medication that they themselves take and suggest to others. Therefore, potential problems of self-medication should be emphasised to the students

Dendritic cell regulation of HIV-1 latency in CD4+ T-cell

Combination antiretroviral therapy (cART) is able to control HIV-1 replication and prevent acquired immune deficiency syndrome (AIDS), however does not provide a cure. The major barrier to HIV-1 cure is the persistence of long-lived, latently infected resting CD4+ T-cells. We developed an in vitro dendritic cell (DC)-T-cell latency model, which showed that myeloid (m)DC induced post-integration latent infection in non-proliferating, resting and proliferating CD4+ T-cells. Gene expression profiles indicated that mDC could regulate the establishment of latent infection in non-proliferating and proliferating CD4+ T-cells through interaction between adhesion molecules, co-stimulatory molecules and immune checkpoints. On the contrary, pDC-T-cell interactions led to the inhibition of latent infection in co-cultured resting CD4+ T-cells and death of proliferated CD4+ T-cells. Gene expression profiles suggested that inhibition is mediated through type-I and III interferons. Investigation of the mechanisms of mDC mediated establishment of latent infection and pDC mediated inhibition of latent infection, may provide novel elimination strategies for HIV-1 cure

A quality initiative to improve exclusive breast milk feeding in preterm neonates

Background and Objective: Preterm babies thrive well on exclusive breastfeeding with fewer complications. In a retrospective analysis, challenges were noticed in the form of delayed first milk expression and frequency of expression, which was limited to 2–3 times a day. A quality improvement initiative was designed to increase the exclusive breast milk feeding rates in admitted preterm babies. The purpose of the present study is to evaluate the proportion of preterm neonates receiving mother's own milk by day 7 of life after such an initiative. Methods: The quality improvement initiative was conducted at a level 3 neonatal intensive care unit from May 10, 2018, to August 10, 2018. Inborn preterm neonates <34 weeks with a minimum hospital stay of 7 days were included. Neonates of critically sick mothers and neonates having major congenital malformations were excluded. Mothers were given structured antenatal counseling regarding expressing breast milk (EBM). Postnatal visits were conducted to provide information through a video and a leaflet and formation of breastfeeding support group. Results: A total of 30 babies were recruited during a period of 3 months and compared to historical controls. The median age of the first EBM improved from 2.5 days to 1.16 days, and the amount of EBM on the first day improved from 0.24 ml to 2.6 ml (p = .002). The proportion of babies receiving EBM on the first day improved from 24% to 80% (p = .0001), and at the end of 7 days, it increased to 73% from 46% (p = .02). The factors time to reach full enteral feeds, time to regain birth weight, rate of necrotizing enterocolitis, rate of sepsis, and proportion of babies on exclusive mother's milk during discharge appear comparable in both the groups. Conclusions: A simple quality improvement approach through the formation of breastfeeding support group, antenatal counseling, and postnatal support for breast milk expression has made a significant improvement in milk expression from mothers delivered prematurely. It reflects on a team approach using the available resources. Keywords: Exclusive breastfeeding, Preterm neonate, Quality improvemen

Problems and solutions to conduct of thesis of postgraduate medical students during the COVID-19 pandemic: an insight into the students perspective

Purpose The coronavirus disease 2019 (COVID-19) pandemic has adversely impacted medical education worldwide. However, its impact on the postgraduate medical thesis and dissertation work is still not evaluated. Through this study, we planned to find out the problems brought by the pandemic and likely alternatives and possible solutions to thrust the academic competence of postgraduate students. Methods After obtaining institutional ethics committee approval, we sent a 13-item questionnaire to postgraduate medical students in India via various social media online platforms. Data on the impact of the COVID-19 pandemic on thesis work and alternatives/solutions to improve the research competence were collected on a Likert scale and analyzed. Results We received a total of 398 responses out of which 377 entries were included for final analysis. The majority of participants (88%) reportedly had an adverse impact on the thesis work and out of 25% of the participants who recently submitted their around 45% had to do so without achieving the estimated sample size. The 6-month departmental review for thesis progress was seen in merely 28% of participants. Possible alternatives suggested were the maintenance of log books, task-based assessment of research methodology, departmental audits, and systematic reviews. Solutions suggested for improving the research competence of students were a compulsory research methodology curriculum, a biostatistics department in each institution, permission to conduct thesis work beyond submission time, exclusive time for research work, and financial incentives. Conclusion Modification in the research aspect of the current postgraduate medical education is the need of the hour and the pandemic has enlightened us regarding the current weaknesses

Forest policies and programs affecting vulnerability and adaptation to climate change

Due to large scale afforestation programs and forest conservation legislations, India's total forest area seems to have stabilized or even increased. In spite of such efforts, forest fragmentation and degradation continues, with forests being subject to increased pressure due to anthropogenic factors. Such fragmentation and degradation is leading to the forest cover to change from very dense to moderately dense and open forest and 253 km(2) of very dense forest has been converted to moderately dense forest, open forest, scrub and non-forest (during 2005-2007). Similarly, there has been a degradation of 4,120 km(2) of moderately dense forest to open forest, scrub and non-forest resulting in a net loss of 936 km(2) of moderately dense forest. Additionally, 4,335 km(2) of open forest have degraded to scrub and non-forest. Coupled with pressure due to anthropogenic factors, climate change is likely to be an added stress on forests. Forest sector programs and policies are major factors that determine the status of forests and potentially resilience to projected impacts of climate change. An attempt is made to review the forest policies and programs and their implications for the status of forests and for vulnerability of forests to projected climate change. The study concludes that forest conservation and development policies and programs need to be oriented to incorporate climate change impacts, vulnerability and adaptation

Novel RT-ddPCR Assays for determining the transcriptional profile of SARS-CoV-2

The exact mechanism of coronavirus replication and transcription is not fully understood; however, a hallmark of coronavirus transcription is the generation of negative-sense RNA intermediates that serve as the templates for the synthesis of positive-sense genomic RNA (gRNA) and an array of subgenomic mRNAs (sgRNAs) encompassing sequences arising from discontinuous transcription. Existing PCR-based diagnostic assays for SAR-CoV-2 are qualitative or semi-quantitative and do not provide the resolution needed to assess the complex transcription dynamics of SARS-CoV-2 over the course of infection. We developed and validated a novel panel of specially designed SARS-CoV-2 ddPCR-based assays to map the viral transcription profile. Application of these assays to clinically relevant samples will enhance our understanding of SARS-CoV-2 replication and transcription and may also inform the development of improved diagnostic tools and therapeutics. Highlights We developed a novel panel of 7 quantitative RT-ddPCRs assays for SARS-Cov-2 Our panel targets nongenic and genic regions in genomic and subgenomic RNAs All assays detect 1-10 copies and are linear over 3-4 orders of magnitude All assays correlated with the clinical Abbott SARS-CoV-2 Viral Load Assay Clinical samples showed higher copy numbers for targets at the 3’ end of the genom

Cardiac output monitoring: Technology and choice

The accurate quantification of cardiac output (CO) is given vital importance in modern medical practice, especially in high-risk surgical and critically ill patients. CO monitoring together with perioperative protocols to guide intravenous fluid therapy and inotropic support with the aim of improving CO and oxygen delivery has shown to improve perioperative outcomes in high-risk surgical patients. Understanding of the underlying principles of CO measuring devices helps in knowing the limitations of their use and allows more effective and safer utilization. At present, no single CO monitoring device can meet all the clinical requirements considering the limitations of diverse CO monitoring techniques. The evidence for the minimally invasive CO monitoring is conflicting; however, different CO monitoring devices may be used during the clinical course of patients as an integrated approach based on their invasiveness and the need for additional hemodynamic data. These devices add numerical trend information for anesthesiologists and intensivists to use in determining the most appropriate management of their patients and at present, do not completely prohibit but do increasingly limit the use of the pulmonary artery catheter