ASXL1 mutations are associated with distinct epigenomic alterations that lead to sensitivity to venetoclax and azacytidine.

The BCL2-inhibitor, Venetoclax (VEN), has shown significant anti-leukemic efficacy in combination with the DNMT-inhibitor, Azacytidine (AZA). To explore the mechanisms underlying the selective sensitivity of mutant leukemia cells to VEN and AZA, we used cell-based isogenic models containing a common leukemia-associated mutation in the epigenetic regulator ASXL1. KBM5 cells with CRISPR/Cas9-mediated correction of the ASXL1G710X mutation showed reduced leukemic growth, increased myeloid differentiation, and decreased HOXA and BCL2 gene expression in vitro compared to uncorrected KBM5 cells. Increased expression of the anti-apoptotic gene, BCL2, was also observed in bone marrow CD34+ cells from ASXL1 mutant MDS patients compared to CD34+ cells from wild-type MDS cases. ATAC-sequencing demonstrated open chromatin at the BCL2 promoter in the ASXL1 mutant KBM5 cells. BH3 profiling demonstrated increased dependence of mutant cells on BCL2. Upon treatment with VEN, mutant cells demonstrated increased growth inhibition. In addition, genome-wide methylome analysis of primary MDS samples and isogenic cell lines demonstrated increased gene-body methylation in ASXL1 mutant cells, with consequently increased sensitivity to AZA. These data mechanistically link the common leukemia-associated mutation ASXL1 to enhanced sensitivity to VEN and AZA via epigenetic upregulation of BCL2 expression and widespread alterations in DNA methylation.

Comparison between Epidural Ropivacaine versus Ropivacaine with Clonidine in Patients Undergoing Abdominal Hysterectomy: A Randomized Study.

CONTEXT: Regional anesthesia has emerged as one of the preferred and convenient modes for intra- and post-operative management owing to its advantage of not interfering with the metabolic functions, better tolerability, and decrease in reflex activity. In recent years, ropivacaine has increasingly replaced bupivacaine as a preferred local anesthetic because of its similar analgesic properties, lesser motor blockade, and decreased propensity of cardiotoxicity. Neuraxial adjuvant such as clonidine used in epidural anesthesia offers advantage by augmenting the local anesthetic effect and reducing the anesthetic and analgesic requirement. AIMS AND OBJECTIVES: Comparison of onset, duration of sensory and motor block, and any adverse effects between 0.5% ropivacaine with normal saline versus 0.5% ropivacaine with clonidine (75 µg/kg). MATERIALS AND METHODS: This prospective randomized study was carried out in 50 patients (25 in each group) of American Society of Anesthesiologist Grade 1 and 2 scheduled for abdominal hysterectomy under epidural block. Group-1 (ropivacaine-clonidine [RC]): Epidural ropivacaine 20 ml (0.5%) with 0.75 µg/kg clonidine. Group-2 (ropivacaine [R]): Epidural ropivacaine 20 ml (0.5%) with normal saline. Onset, duration of sensory-motor block, heart rate, blood pressure, oxygen saturation, and respiratory rate were recorded. STATISTICAL ANALYSIS: The statistical analysis was done using Statistical Package for Social Sciences version 15.0. Chi-square test, ANOVA, Student's t-test, and paired t-test were used. RESULTS: Groups were comparable with regard to demographic data and hemodynamic stability. Onset of sensory and motor blockade was faster in RC group as compared to R group. Duration of postoperative analgesia was significantly prolonged in RC group. No potential side effect was seen in either group. CONCLUSION: On account of faster onset, hemodynamic stability, and prolonged postoperative analgesia, ropivacaine with clonidine is a better option than ropivacaine alone.

Postoperative Analgesia After Panhysterectomy, Addition of Clonidine to Bupivacaine: Boon for the Patients.

INTRODUCTION: Postoperative period after panhysterectomy is very painful as there is too much tissue handling. In the practice of regional anesthesia neuraxial, opioids have been used extensively as an adjuvant to bupivacaine to enhance the potency and duration of sensory and motor block produced by bupivacaine with satisfactory results. However, delayed respiratory depression by opioids has prompted further research to develop nonopioid analgesics. This study was undertaken to assess the degree of sensory and motor block and postoperative analgesia provided by low dose 50 µg intrathecal clonidine admixed with 0.5% hyperbaric bupivacaine as compared to bupivacaine alone in patients undergoing a total abdominal hysterectomy. MATERIALS AND METHODS: Hundred adult patients of American Society of Anesthesiologist Class 1 and 2 were randomly allocated to Group A and Group B. Group A patients received 15 mg 0.5% hyperbaric bupivacaine with 50 µg clonidine intrathecally. Group B patients received 15 mg 0.5% hyperbaric bupivacaine with normal saline. OBSERVATION AND RESULTS: The mean duration of motor block was significantly higher in Group A (270.80± 66.0 min) as compared to Group B (184.60 ± 72.03 min), with statistically significant difference. There was also statistically significant difference in the duration of sensory block between Group A (290.20 ± 80.27 min) and Group B (190.83 ± 86.90 min). The duration of postoperative analgesia was significantly higher in Group A as compared to Group B (541.06 ± 130.64 min and 252.80 ± 84.10 min respectively). CONCLUSION: Addition of intrathecal clonidine 50 µg to bupivacaine (15 mg, 0.5%) prolongs the duration of sensory and motor block and duration of analgesia, thus produces an effective spinal anesthesia and good postoperative analgesia for longer duration and reduced postoperative analgesic requirement.

Relevance of Haematologic Parameters in Obese Women with or without Metabolic Syndrome.

INTRODUCTION: Obesity is rapidly growing problem worldwide. It predisposes to a variety of serious ailments including heart disease, diabetes mellitus, degenerative joint disease, atherosclerosis, etc. This is probably related to proinflammatory state associated with obesity due to release of several inflammatory mediators by the adipose tissue. The mediators are also probably responsible for metabolic syndrome associated with obesity. Besides, they may also induce significant changes in haematological parameters associated with inflammation. AIM: Present study was undertaken to ascertain the relationship between obesity and leucocyte counts (particularly TLC and ANC) and find out if the changes induced in them are significant enough to be used as predictors of metabolic syndrome. MATERIALS AND METHODS: This case-control study was carried out on 243 female subjects allocated to four groups based on WHO and IDF criteria: Control, Overweight, Obese and Obese with Metabolic Syndrome. From all the subjects, data pertaining to obesity related anthropometric measurements, lipid profile, fasting plasma glucose levels and complete blood counts were collected. These were analysed statistically. RESULTS: There was a strong positive correlation between obesity related anthropometric measurements (BMI, BF, WC) and leucocyte counts - TLC and ANC - which were statistically highly significant; TNC and ANC also showed strong positive correlation with FPG. Mean values for TLC and ANC showed statistically significant difference between each and every group. The difference in the mean values of these parameters between obese and metabolic syndrome was highly significant. Both elevated FPG and BMI were independently associated with relative leucocytosis; when both of them were elevated simultaneously, the effect appeared to be potentiating. CONCLUSION: Increase in obesity associated anthropometric measurements (BMI, WC, BF) is associated with relative leucocytosis within the physiological range. The changes in TLC and ANC are significant enough to be used as predictors of onset of metabolic syndrome in obese subjects.

Germinoma-unusual presentation: a case report.

Germinoma accounts for two-thirds of germ cell tumors and about 40% of all pineal region neoplasms. This case illustrates an unusual manifestation of metatastic germinoma with spread to ventricles and meninges without a pineal mass. A 24-year-old man presented with nausea, vertigo, and left facial droop. Cerebrospinal fluid aspirate showed malignant cells most suggestive of a germinoma.

Aquaporin 5 regulates cigarette smoke induced emphysema by modulating barrier and immune properties of the epithelium.

Chronic obstructive pulmonary disease (COPD) causes significant morbidity and mortality. Cigarette smoke, the most common risk factor for COPD, induces airway and alveolar epithelial barrier permeability and initiates an innate immune response. Changes in abundance of aquaporin 5 (AQP5), a water channel, can affect epithelial permeability and immune response after cigarette smoke exposure. To determine how AQP5-derived epithelial barrier modulation affects epithelial immune response to cigarette smoke and development of emphysema, WT and AQP5(-/-) mice were exposed to cigarette smoke (CS). We measured alveolar cell counts and differentials, and assessed histology, mean-linear intercept (MLI), and surface-to-volume ratio (S/V) to determine severity of emphysema. We quantified epithelial-derived signaling proteins for neutrophil trafficking, and manipulated AQP5 levels in an alveolar epithelial cell line to determine specific effects on neutrophil transmigration after CS exposure. We assessed paracellular permeability and epithelial turnover in response to CS. In contrast to WT mice, AQP5(-/-) mice exposed to 6 months of CS did not demonstrate a significant increase in MLI or a significant decrease in S/V compared with air-exposed mice, conferring protection against emphysema. After sub-acute (4 weeks) and chronic (6 mo) CS exposure, AQP5(-/-) mice had fewer alveolar neutrophil but similar lung neutrophil numbers as WT mice. The presence of AQP5 in A549 cells, an alveolar epithelial cell line, was associated with increase neutrophil migration after CS exposure. Compared with CS-exposed WT mice, neutrophil ligand (CD11b) and epithelial receptor (ICAM-1) expression were reduced in CS-exposed AQP5(-/-) mice, as was secreted LPS-induced chemokine (LIX), an epithelial-derived neutrophil chemoattractant. CS-exposed AQP5(-/-) mice demonstrated decreased type I pneumocytes and increased type II pneumocytes compared with CS-exposed WT mice suggestive of enhanced epithelial repair. Absence of AQP5 protected against CS-induced emphysema with reduced epithelial permeability, neutrophil migration, and altered epithelial cell turnover which may enhance repair.

Disparity in survival outcome after hematopoietic stem cell transplantation for hematologic malignancies according to area of primary residence.

We evaluated whether or not a patient's area of primary residence is an independent risk factor for overall survival (OS) after HLA-identical sibling or autologous hematopoietic stem cell transplantation (HSCT). This retrospective cohort study included patients who underwent autologous (n = 1739) or HLA-identical sibling (n = 267) HSCT to treat a hematologic malignancy between 1983 and 2004 at the University of Nebraska Medical Center. Primary area of residence, using the patient's zip code, was categorized as either urban or rural (including isolated, small rural, or large rural) according to the Rural Urban Commuting Area Codes (RUCA) classification system. An association between area of primary residence and survival was examined using Cox proportional hazards regression analysis while adjusting for patient-, disease-, and treatment-related variables. Patients from rural areas who received autologous HSCT had a higher relative risk of death (relative risk = 1.18; P = .016) than urban patients who underwent the same procedure. Survival rates in patients from rural and urban locations are as follows: 1 year, 73% vs 78% (P = .04); 5 year, 48% vs 54% (P = .012). We failed to detect a significant difference in the risk of death according to primary area of residence in the HLA-identical sibling HSCT cohort, although this may be from lack of statistical power. Our findings suggest that the primary location of a patient's residence may be an independent risk factor for survival after HSCT.