Robertsonfrazier7786

Human kinome/phosphatome screen identified CAMK2N1 genes suppressing the development of human hepatocellular carcinoma (HCC). CAMK2N1 downregulation was found in 47% HCCs and associated with poor prognosis. The downregulation was mainly attributed to its genome deletion (28.4%) and DNA hypermethylation of its promoter (12.5%). Silencing and ectopic expression of CAMK2N1 respectively enhanced and suppressed cell proliferation, colony formation, and xenograft tumor growth in nude mice. Comparative proteomics revealed that CAMK2N1 silencing transcriptionally deregulated the genes regulated by E2F1 (89 out of the 114 E2F-signaling targets, P = 8.8E-240). The promoter assays revealed that CAMK2N1 suppressed E2F1-mediated transcriptional activities. CAMK2N1 silencing induced cyclins D/E expression, whereas its ectopic expression induced P27/KIP1 expression and suppressed the cell cycle. CAMK2N1 was translocated from the nuclei to the cytoplasm when cell proliferation reached the stationary phase, where its functions as an endogenous inhibitor of CAMK2. In conclusion, CAMK2NA is a novel 1p36 tumor suppressor gene that inhibits E2F1 transcriptional activities and induces P27/KIP1 expression. CAMK2N1-CAMK2 signaling forms a mechanism that restricts the cell cycle progression. Its deregulation could lead to tumorigenesis and might serve as promising therapeutic targets.Electron cryo-microscopy (cryoEM) involves the estimation of spatial rotations, or saying orientations, of projection images or three-dimensional (3D) volumes. Euler angle system is widely used to describe spatial rotations in most cryoEM algorithms and software. In this review, we introduce unit quaternion as an alternate to Euler angles for describing spatial rotations, customize and develop corresponding tools for increasing demands of statistical analysis of spatial rotations in cryoEM. Some basic properties and definitions of quaternion are first recalled. Thereafter, distance and geodesic between rotations are introduced to aid comparisons and interpolations between rotations, which are prerequisites of statistics of rotations in 3D cryoEM. Furthermore, statistics of rotations are reviewed. Techniques potentially useful in cryoEM, such as calculations of the average rotation, generation of quasi-regular grids, sampling, inference with uniform distribution and angular central Gaussian (ACG) distribution, and estimation of rotation precision, are reviewed and developed. Finally, molecular symmetry presented in unit quaternion form is discussed. Unit quaternion system is shown as a convenient and comprehensive mathematical tool for cryoEM.The objective of the present investigation was to understand the impact of exogenously applied melatonin on mitochondrial respiration and sugar metabolism in two contrasting rice cultivars, viz., Khitish (arsenic-susceptible) and Muktashri (arsenic-tolerant) under arsenic-stress. Melatonin effectively restored the level of organic acids like pyruvic acid, malic acid and more particularly citric acid by 33 % in Khitish which were lowered during arsenic-stress, whereas their levels were further elevated in Muktashri to provide energy for defence against arsenic-induced injury. Arsenic-exposure led to a significant inhibition in enzyme activities as well as corresponding transcript level of key respiratory enzymes, viz., pyruvate dehydrogenase, citrate synthase, isocitrate dehydrogenase, succinate dehydrogenase and malate dehydrogenase, intriguingly more prominently in case of Khitish. Conversely, melatonin supplementation, irrespective of cultivars, considerably improved the activity of the above enzymes and corresponding gene expressions during stress, indicating acceleration in the rate of Krebs cycle. Melatonin supplementation also stimulated the accumulation of total soluble sugars by 62 % and 25 %, reducing sugars by 50 % and 44 % and non-reducing sugars by 75 % and 14 % in Khitish and Muktashri respectively, concomitant with higher activities of acid invertase, sucrose synthase and sucrose phosphate synthase enzymes, along with the expression of corresponding genes. Enhanced starch accumulation via regulation of alpha amylase and starch phosphorylase activities and gene expression, by melatonin also contributed towards better stress tolerance. Overall, this work illustrated the efficacy of melatonin in the regulation of representative organic acids and enzymes of respiratory cycle along with starch and sugar metabolism in rice cultivars under arsenic toxicity.Respiratory syncytial virus (RSV) is a highly contagious virus causing severe infection in infants and the elderly. Various approaches are being used to develop an effective RSV vaccine. The RSV fusion (F) subunit, particularly the cleaved trimeric pre-fusion F, is one of the most promising vaccine candidates under development. learn more -fusion conformation elicits the majority of neutralizing antibodies during natural infection. However, this pre-fusion conformation is metastable and prone to conversion to a post-fusion conformation, thus hindering the potential of this construct as a vaccine antigen. The Vaccine Research Center (VRC) at the National Institutes of Health (NIH) designed a structurally stabilized pre-fusion F glycoprotein, DS-Cav1, that showed high immunogenicity and induced a neutralizing response in animal studies. To advance this candidate to clinical manufacturing, a production process that maintained product quality (i.e. a cleaved trimer with pre-fusion conformation) and delivered high protein expression levels was required. #link# This report describes the development of the vaccine candidate including vector design and cell culture process development to meet these challenges. Co-transfection of individual plasmids to express DS-Cav1 and furin (for DS-Cav1 cleavage and activation) demonstrated a superior protein product expression and pre-fusion conformation compared to co-expression with a double gene vector. A top clone was selected based on these measurements. Protein expression levels were further increased by seeding density optimization and a biphasic hypothermia temperature downshift. The combined efforts led to a high-yield fed-batch production of approximately 1,500 mg/L (or up to 15,000 doses per liter) at harvest. The process was scaled up and demonstrated to be reproducible at 50 L-scale for toxicity and Phase I clinical trial use. Preliminary phase I data indicate the pre-fusion antigen has a promising efficacy (Crank et al., 2019).