Application of Ketamine in Current Practice of Anesthesiology

Ketamine was discovered in 1964 by merging a ketone with an amine. Patients described feeling disconnected like they were floating in outer. Thus, it was characterized as a dissociative anesthetic. It is a unique drug that expresses hypnotic, analgesic, and amnesic effects. No other drug used in clinical practice produces these three important effects at the same time. Its newly found neuroprotective, anti-inflammatory, antitumor effects and low dose applications have helped to widen the clinical profile of ketamine. Ketamine as an analgesic adjunct in chronic pain patients is currently being researched. Combined use of ketamine and an opiate analgesic has been found to provide good perioperative pain control with reduction in symptoms such as nausea and vomiting, sedation, and respiratory insufficiency

AGNet: Weighing Black Holes with Deep Learning

Supermassive black holes (SMBHs) are ubiquitously found at the centers of most massive galaxies. Measuring SMBH mass is important for understanding the origin and evolution of SMBHs. However, traditional methods require spectroscopic data which is expensive to gather. We present an algorithm that weighs SMBHs using quasar light time series, circumventing the need for expensive spectra. We train, validate, and test neural networks that directly learn from the Sloan Digital Sky Survey (SDSS) Stripe 82 light curves for a sample of $38,939$ spectroscopically confirmed quasars to map out the nonlinear encoding between SMBH mass and multi-color optical light curves. We find a 1$\sigma$ scatter of 0.37 dex between the predicted SMBH mass and the fiducial virial mass estimate based on SDSS single-epoch spectra, which is comparable to the systematic uncertainty in the virial mass estimate. Our results have direct implications for more efficient applications with future observations from the Vera C. Rubin Observatory. Our code, \textsf{AGNet}, is publicly available at \url{https://github.com/snehjp2/AGNet}.Comment: 8 pages, 7 figures, 1 table, Accepted by MNRA

HTLV-1 Tax Mediated Downregulation of miRNAs Associated with Chromatin Remodeling Factors in T Cells with Stably Integrated Viral Promoter

RNA interference (RNAi) is a natural cellular mechanism to silence gene expression and is predominantly mediated by microRNAs (miRNAs) that target messenger RNA. Viruses can manipulate the cellular processes necessary for their replication by targeting the host RNAi machinery. This study explores the effect of human T-cell leukemia virus type 1 (HTLV-1) transactivating protein Tax on the RNAi pathway in the context of a chromosomally integrated viral long terminal repeat (LTR) using a CD4+ T-cell line, Jurkat. Transcription factor profiling of the HTLV-1 LTR stably integrated T-cell clone transfected with Tax demonstrates increased activation of substrates and factors associated with chromatin remodeling complexes. Using a miRNA microarray and bioinformatics experimental approach, Tax was also shown to downregulate the expression of miRNAs associated with the translational regulation of factors required for chromatin remodeling. These observations were validated with selected miRNAs and an HTLV-1 infected T cells line, MT-2. miR-149 and miR-873 were found to be capable of directly targeting p300 and p/CAF, chromatin remodeling factors known to play critical role in HTLV-1 pathogenesis. Overall, these results are first in line establishing HTLV-1/Tax-miRNA-chromatin concept and open new avenues toward understanding retroviral latency and/or replication in a given cell type

Down regulation of miR-149 and miR-873 in HTLV-1 infected cell line, MT-2.

<p>miR-873 were validated for their presence in MT-2 and Jurkat cells by real time PCR. Amplification plots were obtained for the same and plotted with their respective Ct values in both the cell lines with GAPDH as internal control (A). Fold-change in the expression was calculated based on the ΔΔCt method. Triplicate samples were utilized to derive the standard deviation and significance by t-test represented here as * for p<0.05.</p

Verifying the functionality of the integrated HTLV-1 LTR-luc by determining the ability to respond to Tax.

<p>The stably integrated pooled clone (clones C1, C2, and C4) was compared to transiently transfected HTLV-1 LTR-luc Jurkat with respect to its ability to respond to Tax by transiently transfecting both cell lines with either pCMV (control) or pCMV-Tax. Luciferase activity was measured 24 h post-transfection. (A) Stably integrated pooled clone versus the control clone (without integrated viral promoter). (B) Transiently transfected Jurkat clone with pU3R-luc versus control Jurkat (without transfected viral promoter). Each value shown represents the average of duplicate transfection reactions and is representative of two independent experiments. Error bars indicate the standard deviation of one representative experiment.</p

Screening of HTLV-1 LTR-luc stable integrants.

<p>(A) Luciferase activity in stably integrated HTLV-1 LTR Jurkat clones was measured in terms of relative light units. Control represents the luciferase activity from the cells that were stably integrated with pGL3-basic vector and pRSV-neo. Each value shown represents the average of duplicate transfection reactions and is representative of at least two independent experiments. Error bars indicate standard deviation (±) from one representative experiment. (B) For verifying HTLV-1 LTR-luc integration, all five clones (C1–C5) were examined and confirmed qualitatively for HTLV-1 LTR-luc integration for both LTR and luciferase genes independently by PCR amplification of LTR with product size 183 bp and Southern blot hybridization using luciferase probe following PCR amplification of luciferase gene with product size 1.6 kb. The control clone represents the Jurkat clone stably integrated with pGL3-basic vector and pRSV-neo in the absence of the HTLV-1 LTR. The positive control in each case is the pU3R-luc plasmid. (D) For quantifying HTLV-1 provirus copy numbers by real-time PCR in the stably integrated clones, all five clones (C1–C5) were selected for real-time PCR analysis to determine the copy number of copies of the integrated LTR per 100 cells. Standard curves for β-actin and the HTLV-1 LTR were generated from Jurkat and TARL-2 cell lines, respectively, with the coefficient of correlation, <i>r²</i>, being greater than 0.996 for both.</p

Mechanism to explain the differential requirements of cellular chromatin remodeling factors in stably integrated versus transiently transfected HTLV-1 LTR.

<p>The major differences in the need for cellular chromatin remodeling factors between a stably integrated and transiently transfected viral promoter are outlined. In the case of a stably integrated LTR, Tax mediates the increased recruitment of the chromatin remodeling factors (HATs and SWI/SNF) and their substrates. In a transient transfection, the LTR is more readily accessible and thus exhibits a reduced need for such factors.</p

Effect of miRNA mimics on the expression of p300 and p/CAF in MT-2 cells.

<p>(A) Equal protein amounts from the whole cell lysates of MT-2 and Jurkat cells lines were resolved on SDS polyacrylamide gel and then transferred to a PVDF membrane. The membrane was then incubated with anti-p300 (1∶1000 dilution) and anti-p/CAF (1∶1000 dilution) monoclonal antibodies for 2 h at room temperature and incubated for 1 h with an anti-rabbit secondary antibody conjugated to IRDy800 and IRDy680, respectively. Signals were detected by Licor Pearl Pulse Imager. It was observed that the levels of p300 and p/CAF went down on transfection of MT-2 cells with respective miR-mimic when compared to non-transfected MT-2 cells. (B) HTLV-1 p19 was quantitated with ELISA in MT-2 culture supernatant and compared to supernatant obtained from MT-2 cells transfected with miR-149 and miR-873 mimics.</p