Cardiac Computed Tomography Angiography in Infants and Young Children Without Sedation.

Cardiac computed tomography (CT) imaging plays a pivotal role in the diagnosis and management of infants and young children with congenital heart disease (CHD). While the benefits of CT imaging are well-established, the challenge lies in adapting these procedures to the unique requirements of infants and young children. Traditionally, sedation has been a common practice to ensure cooperation and motion control during imaging. However, using sedation introduces its challenges including potential risks, limitations, and cost implications. In this study, authors explore the feasibility, safety, and diagnostic accuracy of unsedated cardiac CT examinations in infants and young children. This study proves cardiac CT can be performed in India without sedation using simple restraining techniques. This approach aligns with the cultural and familial dynamics prevalent in the country and holds the potential to address economic and infrastructure challenges.

A guide to epigenetics in leukaemia stem cells.

Leukaemia stem cells (LSCs) are the critical seed for the growth of haematological malignancies, driving the clonal expansion that enables disease initiation, relapse and often resistance. Specifically, they display inherent phenotypic and epigenetic plasticity resulting in complex heterogenic diseases. In this review, we discuss the key principles of deregulation of epigenetic processes that shape this disease evolution. We consider measures to define and quantify clonal heterogeneity, combining information from recent studies assessing mutational, transcriptional and epigenetic landscapes at single cell resolution in myeloid neoplasms (MN). We highlight the importance of integrating epigenetic and genetic information to better understand inter- and intra-patient heterogeneity and discuss how this understanding further informs evolution and progression trajectories and subsequent clinical response in MN. Under this topic, we also discuss efforts to identify mechanisms of resistance, by longitudinal analyses of patient samples. Finally, we highlight how we might target these aberrant epigenetic processes for better therapeutic outcomes and to potentially eradicate LSCs.

Production of Designer Xylose-Acetic Acid Enriched Hydrolysate from Bioenergy Sorghum, Oilcane, and Energycane Bagasses.

Xylan accounts for up to 40% of the structural carbohydrates in lignocellulosic feedstocks. Along with xylan, acetic acid in sources of hemicellulose can be recovered and marketed as a commodity chemical. Through vibrant bioprocessing innovations, converting xylose and acetic acid into high-value bioproducts via microbial cultures improves the feasibility of lignocellulosic biorefineries. Enzymatic hydrolysis using xylanase supplemented with acetylxylan esterase (AXE) was applied to prepare xylose-acetic acid enriched hydrolysates from bioenergy sorghum, oilcane, or energycane using sequential hydrothermal-mechanical pretreatment. Various biomass solids contents (15 to 25%, w/v) and xylanase loadings (140 to 280 FXU/g biomass) were tested to maximize xylose and acetic acid titers. The xylose and acetic acid yields were significantly improved by supplementing with AXE. The optimal yields of xylose and acetic acid were 92.29% and 62.26% obtained from hydrolyzing energycane and oilcane at 25% and 15% w/v biomass solids using 280 FXU xylanase/g biomass and AXE, respectively.

HOXA9 forms a repressive complex with nuclear matrix-associated protein SAFB to maintain acute myeloid leukemia.

HOXA9 is commonly upregulated in acute myeloid leukemia (AML), in which it confers a poor prognosis. Characterizing the protein interactome of endogenous HOXA9 in human AML, we identified a chromatin complex of HOXA9 with the nuclear matrix attachment protein SAFB. SAFB perturbation phenocopied HOXA9 knockout to decrease AML proliferation, increase differentiation and apoptosis in vitro, and prolong survival in vivo. Integrated genomic, transcriptomic, and proteomic analyses further demonstrated that the HOXA9-SAFB (H9SB)-chromatin complex associates with nucleosome remodeling and histone deacetylase (NuRD) and HP1γ to repress the expression of factors associated with differentiation and apoptosis, including NOTCH1, CEBPδ, S100A8, and CDKN1A. Chemical or genetic perturbation of NuRD and HP1γ-associated catalytic activity also triggered differentiation, apoptosis, and the induction of these tumor-suppressive genes. Importantly, this mechanism is operative in other HOXA9-dependent AML genotypes. This mechanistic insight demonstrates the active HOXA9-dependent differentiation block as a potent mechanism of disease maintenance in AML that may be amenable to therapeutic intervention by targeting the H9SB interface and/or NuRD and HP1γ activity.

Mutational synergy during leukemia induction remodels chromatin accessibility, histone modifications and three-dimensional DNA topology to alter gene expression.

Altered transcription is a cardinal feature of acute myeloid leukemia (AML); however, exactly how mutations synergize to remodel the epigenetic landscape and rewire three-dimensional DNA topology is unknown. Here, we apply an integrated genomic approach to a murine allelic series that models the two most common mutations in AML: Flt3-ITD and Npm1c. We then deconvolute the contribution of each mutation to alterations of the epigenetic landscape and genome organization, and infer how mutations synergize in the induction of AML. Our studies demonstrate that Flt3-ITD signals to chromatin to alter the epigenetic environment and synergizes with mutations in Npm1c to alter gene expression and drive leukemia induction. These analyses also allow the identification of long-range cis-regulatory circuits, including a previously unknown superenhancer of Hoxa locus, as well as larger and more detailed gene-regulatory networks, driven by transcription factors including PU.1 and IRF8, whose importance we demonstrate through perturbation of network members.

Association of Leptin and Carotid Intima-Media Thickness in Overweight and Obese Individuals: A Cross-sectional Study.

OBJECTIVE: Leptin levels are increased in obesity and have been found to be strongly associated with obesity, increased risk of cardiovascular diseases and morbidity. While, carotid intima-media thickness (CIMT) is measured to predict atherosclerosis in early phase. Thus, the objective of this study was to evaluate the leptin levels and CIMT in overweight and obese individuals. METHODS: This cross-sectional study involving 95 subjects, was performed over a period of 1 year in the Department of Medicine, King George's Medical University, Lucknow. Anthropometric measurements included weight, height, waist circumference (WC), and BMI (Kg/m2). Baseline investigations were fasting blood glucose and lipid profile. Quantitative estimation of leptin was done by leptin ELISA, and CIMT was measured using a high-resolution B-mode ultrasound scanner with a 7 MHz linear transducer. Unpaired t-test or ANNOVA was used to compare quantitative variables, and chi-square or fisher's exact test was used to compare categorical variables. Pearson's correlation coefficient was used to test the strength of correlation. A p value of < 0.05 was considered as statistically significant. RESULTS: Based on both BMI and WC, mean leptin levels were significantly increased in overweight and obese subjects (p < 0.05) as compared to normal subjects. Similarly, based on both BMI and WC, significantly higher proportion of overweight and obese subjects had increased CIMT values (p < 0.05), as compared to normal subjects. Mean leptin levels were positively and significantly correlated with weight, WC, BMI, total cholesterol, triglyceride, LDL-cholesterol, and CIMT. Similarly, CIMT values were positively and significantly correlated with age, weight, WC, BMI, total cholesterol, triglyceride, HDL-cholesterol. CONCLUSION: Findings of this study indicates that overweight and obesity results in significant increase in both leptin levels and CIMT values. Furthermore, increased leptin levels and CIMT values are positively correlated with increase in BMI and WC.

Histone deacetylase 4 promotes type I interferon signaling, restricts DNA viruses, and is degraded via vaccinia virus protein C6.

Interferons (IFNs) represent an important host defense against viruses. Type I IFNs induce JAK-STAT signaling and expression of IFN-stimulated genes (ISGs), which mediate antiviral activity. Histone deacetylases (HDACs) perform multiple functions in regulating gene expression and some class I HDACs and the class IV HDAC, HDAC11, influence type I IFN signaling. Here, HDAC4, a class II HDAC, is shown to promote type I IFN signaling and coprecipitate with STAT2. Pharmacological inhibition of class II HDAC activity, or knockout of HDAC4 from HEK-293T and HeLa cells, caused a defective response to IFN-α. This defect in HDAC4-/- cells was rescued by reintroduction of HDAC4 or catalytically inactive HDAC4, but not HDAC1 or HDAC5. ChIP analysis showed HDAC4 was recruited to ISG promoters following IFN stimulation and was needed for binding of STAT2 to these promoters. The biological importance of HDAC4 as a virus restriction factor was illustrated by the observations that (i) the replication and spread of vaccinia virus (VACV) and herpes simplex virus type 1 (HSV-1) were enhanced in HDAC4-/- cells and inhibited by overexpression of HDAC4; and (ii) HDAC4 is targeted for proteasomal degradation during VACV infection by VACV protein C6, a multifunctional IFN antagonist that coprecipitates with HDAC4 and is necessary and sufficient for HDAC4 degradation.

PLZF targets developmental enhancers for activation during osteogenic differentiation of human mesenchymal stem cells.

The PLZF transcription factor is essential for osteogenic differentiation of hMSCs; however, its regulation and molecular function during this process is not fully understood. Here, we revealed that the ZBTB16 locus encoding PLZF, is repressed by Polycomb (PcG) and H3K27me3 in naive hMSCs. At the pre-osteoblast stage of differentiation, the locus lost PcG binding and H3K27me3, gained JMJD3 recruitment, and H3K27ac resulting in high expression of PLZF. Subsequently, PLZF was recruited to osteogenic enhancers, influencing H3K27 acetylation and expression of nearby genes important for osteogenic function. Furthermore, we identified a latent enhancer within the ZBTB16/PLZF locus itself that became active, gained PLZF, p300 and Mediator binding and looped to the promoter of the nicotinamide N-methyltransferase (NNMT) gene. The increased expression of NNMT correlated with a decline in SAM levels, which is dependent on PLZF and is required for osteogenic differentiation.

Comparative Evaluation of Myeloperoxidase Enzymatic Activity in Gingival Crevicular Fluid of Subjects having Orthodontic Treatment by Different Aligning Arch Wires.

INTRODUCTION: There exist a number of factors that affect the outcome of orthodontic treatment. These factors can be assessed by various gingival markers. One such maker is myeloperoxidase (MPO). Hence, we planned the present study to assess and compare the MPO activity in the gingival crevicular fluid (GCF) of subjects undergoing orthodontic treatment by different aligning arch wires. MATERIALS AND METHODS: The present study included assessment of patients who underwent orthodontic treatment for crowding of anterior teeth. Diagnostic cast models of all the subjects were made for recording the irregularity index. All the subjects were randomly divided into three study groups with 15 patients in each group based on the type of nickel-titanium (NiTi) arch wires used. A collection of GCF samples was done in all the patients at various time intervals and it was sent to the laboratory for assessment of MPO activity. Activity of the MPO enzyme was expressed in terms of number of units per 100 µL. All the results obtained were compiled and analyzed by Statistical Package for the Social Sciences (SPSS) software. RESULTS: We observed that nonsignificant results were obtained while comparing the mean age and mean gingival score in all the study groups. However, significant results were obtained on comparing the mean MPO enzymatic activity in all the study groups at different time intervals. CONCLUSION: Both superelastic NiTi and heat-activated NiTi generate optimal forces, which are necessary for higher metabolic response of the periodontal ligament. CLINICAL SIGNIFICANCE: In the intimal stages of orthodontic treatment, both superelastic NiTi and heat-activated NiTi wires are superior in leveling and aligning the crowded teeth.

Loss of the histone methyltransferase EZH2 induces resistance to multiple drugs in acute myeloid leukemia.

In acute myeloid leukemia (AML), therapy resistance frequently occurs, leading to high mortality among patients. However, the mechanisms that render leukemic cells drug resistant remain largely undefined. Here, we identified loss of the histone methyltransferase EZH2 and subsequent reduction of histone H3K27 trimethylation as a novel pathway of acquired resistance to tyrosine kinase inhibitors (TKIs) and cytotoxic drugs in AML. Low EZH2 protein levels correlated with poor prognosis in AML patients. Suppression of EZH2 protein expression induced chemoresistance of AML cell lines and primary cells in vitro and in vivo. Low EZH2 levels resulted in derepression of HOX genes, and knockdown of HOXB7 and HOXA9 in the resistant cells was sufficient to improve sensitivity to TKIs and cytotoxic drugs. The endogenous loss of EZH2 expression in resistant cells and primary blasts from a subset of relapsed AML patients resulted from enhanced CDK1-dependent phosphorylation of EZH2 at Thr487. This interaction was stabilized by heat shock protein 90 (HSP90) and followed by proteasomal degradation of EZH2 in drug-resistant cells. Accordingly, inhibitors of HSP90, CDK1 and the proteasome prevented EZH2 degradation, decreased HOX gene expression and restored drug sensitivity. Finally, patients with reduced EZH2 levels at progression to standard therapy responded to the combination of bortezomib and cytarabine, concomitant with the re-establishment of EZH2 expression and blast clearance. These data suggest restoration of EZH2 protein as a viable approach to overcome treatment resistance in this AML patient population.