Associations with health-related quality of life after intracerebral haemorrhage: pooled analysis of INTERACT studies

Background and purpose Limited data exist on health-related quality of life (HRQoL) after intracerebral haemorrhage (ICH). We aimed to determine baseline factors associated with HRQoL among participants of the pilot and main phases of the Intensive Blood Pressure Reduction in Acute Cerebral Haemorrhage Trials (INTERACT 1 and 2). Methods The INTERACT studies were randomised controlled trials of early intensive blood pressure (BP) lowering in patients with ICH (<6hours) and elevated systolic BP (150-220mmHg). HRQoL was determined using the European Quality of Life Scale (EQ-5D) at 90days, completed by patients or proxy responders. Binary logistic regression analyses were performed to identify factors associated with poor overall HRQoL. Results 2756 patients were included. Demographic, clinical and radiological factors associated with lower EQ-5D utility score were age, randomisation outside of China, antithrombotic use, high baseline National Institutes of Health Stroke Scale (NIHSS) score, larger ICH, presence of intraventricular extension and use of proxy responders. High (14) NIHSS score, larger ICH and proxy responders were associated with low scores in all five dimensions of the EQ-5D. The NIHSS score had a strong association with poor HRQoL (p<0.001). Female gender and antithrombotic use were associated with decreased scores in dimensions of pain/discomfort and usual activity, respectively. Conclusions Poor HRQoL was associated with age, comorbidities, proxy source of assessment, clinical severity and ICH characteristics. The strongest association was with initial clinical severity defined by high NIHSS score. Trial registration numbers NCT00226096 and NCT00716079; Post-results.National Health and Medical Research Council of AustraliaSCI(E)ARTICLE170-758

Stability and Structure of RNA Duplexes Containing Isoguanosine and Isocytidine

Isoguanosine (iG) and isocytidine (iC) differ from guanosine (G) and cytidine (C), respectively, in that the amino and carbonyl groups are transposed. The thermodynamic properties of a set of iG, iC containing RNA duplexes have been measured by UV optical melting. It is found that iG−iC replacements usually stabilize duplexes, and the stabilization per iG−iC pair is sequence-dependent. The sequence dependence can be fit to a nearest-neighbor model in which the stabilities of iG−iC pairs depend on the adjacent iG−iC or G−C pairs. For 5‘-CG-3‘/3‘-GC-5‘ and 5‘-GG-3‘/3‘-CC-5‘ nearest neighbors, the free energy differences upon iG−iC replacement are smaller than 0.2 kcal/mol at 37 °C, regardless of the number of replacements. For 5‘-GC-3‘/3‘-CG-5‘, however, each iG−iC replacement adds 0.6 kcal/mol stabilizing free energy at 37 °C. Stacking propensities of iG and iC as unpaired nucleotides at the end of a duplex are similar to those of G and C. An NMR structure is reported for r(CiGCGiCG)2 and found to belong to the A-form family. The structure has substantial deviations from standard A-form but is similar to published NMR and/or crystal structures for r(CGCGCG)2 and 2‘-O-methyl (CGCGCG)2. These results provide benchmarks for theoretical calculations aimed at understanding the fundamental physical basis for the thermodynamic stabilities of nucleic acid duplexes

Investigation of the Structural Basis for Thermodynamic Stabilities of Tandem GU Wobble Pairs: NMR Structures of (rGGAGUUCC)₂ and (rGGAUGUCC)₂^†^,^‡

The symmetric, tandem GU mismatch motifs, and , which only differ in the mismatch order, have an average difference in thermodynamic stability of 2 kcal/mol at 37 °C. Thermodynamic studies of duplexes containing these motifs indicate the effect is largely localized to the mismatches and adjacent base pairs. The three-dimensional structures of two representative duplexes, (rGGAGUUCC)2 and (rGGAUGUCC)2, were determined by two-dimensional NMR and a simulated annealing protocol. Local deviations are similar to other intrahelical GU mismatches with little effect on backbone torsion angles and a slight overtwisting between the base pair 5‘ of the G of the mismatch and the mismatch itself. Comparisons of the resulting stacking patterns along with electrostatic potential maps suggest that interactions between highly negative electrostatic regions between base pairs may play a role in the observed thermodynamic differences

Table1_Differential Activation of TRPM8 by the Stereoisomers of Menthol.docx

The stereoisomers of menthol elicit cooling sensation to various levels. Though the high-resolution three-dimensional structures of the menthol receptor, the transient receptor potential melastatin 8 (TRPM8) ion channels, have been revolved in different states, the menthol-bound state structure is not determined and how the stereoisomers of menthol interact with TRPM8 remains largely elusive. Taking advantage of the identical atom composition but distinct spatial orientation of chemical groups in menthol stereoisomers, we performed thermodynamic mutant cycle analysis (TMCA) with patch-clamp recordings to probe the interaction between these ligands and TRPM8. By comparing (−)-menthol with (+)-neoisomenthol or (+)-neomenthol, we observed that the isopropyl or hydroxyl group in menthol interacts with the S4 or S3 helix in TRPM8, respectively. These interactions were also corroborated in our molecular docking of the stereoisomers, though the predicted structural details in the interactions of these ligands with TRPM8 residues are different. Therefore, we suggest similar molecular mechanisms of TRPM8 activation by the stereoisomers of menthol, while the binding configuration of an individual stereoisomer is varied.</p

Video1_Differential Activation of TRPM8 by the Stereoisomers of Menthol.MP4

The stereoisomers of menthol elicit cooling sensation to various levels. Though the high-resolution three-dimensional structures of the menthol receptor, the transient receptor potential melastatin 8 (TRPM8) ion channels, have been revolved in different states, the menthol-bound state structure is not determined and how the stereoisomers of menthol interact with TRPM8 remains largely elusive. Taking advantage of the identical atom composition but distinct spatial orientation of chemical groups in menthol stereoisomers, we performed thermodynamic mutant cycle analysis (TMCA) with patch-clamp recordings to probe the interaction between these ligands and TRPM8. By comparing (−)-menthol with (+)-neoisomenthol or (+)-neomenthol, we observed that the isopropyl or hydroxyl group in menthol interacts with the S4 or S3 helix in TRPM8, respectively. These interactions were also corroborated in our molecular docking of the stereoisomers, though the predicted structural details in the interactions of these ligands with TRPM8 residues are different. Therefore, we suggest similar molecular mechanisms of TRPM8 activation by the stereoisomers of menthol, while the binding configuration of an individual stereoisomer is varied.</p

Image1_Differential Activation of TRPM8 by the Stereoisomers of Menthol.JPEG

The stereoisomers of menthol elicit cooling sensation to various levels. Though the high-resolution three-dimensional structures of the menthol receptor, the transient receptor potential melastatin 8 (TRPM8) ion channels, have been revolved in different states, the menthol-bound state structure is not determined and how the stereoisomers of menthol interact with TRPM8 remains largely elusive. Taking advantage of the identical atom composition but distinct spatial orientation of chemical groups in menthol stereoisomers, we performed thermodynamic mutant cycle analysis (TMCA) with patch-clamp recordings to probe the interaction between these ligands and TRPM8. By comparing (−)-menthol with (+)-neoisomenthol or (+)-neomenthol, we observed that the isopropyl or hydroxyl group in menthol interacts with the S4 or S3 helix in TRPM8, respectively. These interactions were also corroborated in our molecular docking of the stereoisomers, though the predicted structural details in the interactions of these ligands with TRPM8 residues are different. Therefore, we suggest similar molecular mechanisms of TRPM8 activation by the stereoisomers of menthol, while the binding configuration of an individual stereoisomer is varied.</p

Image_1_TGF-β1 Promotes Autophagy and Inhibits Apoptosis in Breast Cancer by Targeting TP63.tif

BackgroundBreast cancer (BC) is a prevalent female cancer, which has high morbidity and mortality. However, the pathogenesis of BC has not been fully elucidated. Studies have shown that TGF-β1 plays an important role in regulating the balance between autophagy and apoptosis of tumor. We aim to clarify the specific mechanism of autophagy and apoptosis in breast cancer maintaining the tumor microenvironment.MethodsThe clinical characteristics of 850 BC patients were retrieved from the TCGA database. Differentially expressed autophagy-related genes (DEARGs) between tumor and normal tissues were obtained by the Wilcox test. Through Cox proportional hazard regression analysis, the prognostic risk model was constructed and verified by the ROC curve. We used MDC staining, colony formation assay, CCK-8, flow cytometric analysis to confirm the importance of TGF-β1 on the autophagy and apoptosis of breast cancer cells. Furthermore, western blot was performed to determine the relative expression of protein. The Kaplan-Meier Plotter database was utilized to identify the prognostic value of TP63.ResultsWe successfully constructed a prognostic risk model of breast cancer and screened out an autophagy-related prognostic gene -TP63. We predicted that TGF-β1 and TP63 have a binding site in the JASPAR database as expected. Additionally, TGF-β1 promoted autophagy and inhibited apoptosis of breast cancer cells by inhibiting the expression of TP63.ConclusionOur study demonstrated that the molecular mechanism of TGF-β/TP63 signaling in regulating autophagy and apoptosis of breast cancer and provided a potential prognostic marker in breast cancer.</p

Image2_Differential Activation of TRPM8 by the Stereoisomers of Menthol.JPEG

The stereoisomers of menthol elicit cooling sensation to various levels. Though the high-resolution three-dimensional structures of the menthol receptor, the transient receptor potential melastatin 8 (TRPM8) ion channels, have been revolved in different states, the menthol-bound state structure is not determined and how the stereoisomers of menthol interact with TRPM8 remains largely elusive. Taking advantage of the identical atom composition but distinct spatial orientation of chemical groups in menthol stereoisomers, we performed thermodynamic mutant cycle analysis (TMCA) with patch-clamp recordings to probe the interaction between these ligands and TRPM8. By comparing (−)-menthol with (+)-neoisomenthol or (+)-neomenthol, we observed that the isopropyl or hydroxyl group in menthol interacts with the S4 or S3 helix in TRPM8, respectively. These interactions were also corroborated in our molecular docking of the stereoisomers, though the predicted structural details in the interactions of these ligands with TRPM8 residues are different. Therefore, we suggest similar molecular mechanisms of TRPM8 activation by the stereoisomers of menthol, while the binding configuration of an individual stereoisomer is varied.</p

DataSheet1_Olanzapine induces weight gain in offspring of prenatally exposed poly I:C rats by reducing brown fat thermogenic activity.docx

Background: Olanzapine (OLZ) is an antipsychotic with a high risk of metabolic syndrome, and its induced metabolic disturbance may be related to the thermogenic function of brown adipose tissue (BAT). Of note is that schizophrenia itself appears to be associated with a higher incidence of metabolic syndrome. However, whether OLZ affects metabolic disorders by regulating BAT function and its mechanism in animal models of schizophrenia have not been reported.Methods: We induced maternal immune activation (MIA) in pregnant rodents by injection of synthetic double-stranded RNA-poly I:C (a virus-like substance), and rats were injected with poly I:C, 10 mg/kg) or saline on day 13 of gestation. Rat offspring received OLZ (1 mg/kg, tid) or vehicle from adulthood for 28 days, and body weight and food intake were recorded. Morphological alterations of white adipose tissue (WAT) and BAT were analyzed by HE and oil red staining, and expression of BAT-specific marker proteins/genes was detected by western blot and qRT-PCR. In addition, embryonic stem cells C3H10T1/2 were used to direct differentiation into brown-like adipocytes, and C3H10T1/2 cells were treated with OLZ for the differentiation process. The effects of OLZ on brown-like adipocyte differentiation and activity were analyzed using oil red staining, immunofluorescence and flow cytometry.Results: Compared with the Veh (saline) group, the TG, pWAT weight, adipocyte size and liver weight of the Veh (poly I:C) group were significantly increased, suggesting that the offspring of Poly I:C rats had obvious dyslipidemia and lipid accumulation, which were risk factors for metabolic abnormalities such as obesity. In addition, OLZ treatment resulted in altered WAT and BAT morphology in poly I:C or saline exposed offspring, causing lipid accumulation and weight gain and reducing the expression of the BAT-specific marker molecule UCP1 protein/gene. At the same time, OLZ inhibited the directional differentiation and mitochondrial activity of C3H10T1/2 brown-like adipocytes.Conclusion: Poly I:C-elicited MIA and OLZ differentially inhibited BAT activity and mitochondrial biogenesis, leading to weight gain in adult rats, a process involving PPAR-γ/UCP1-related thermogenic proteins.</p

Nuclear Magnetic Resonance Spectroscopy and Molecular Modeling Reveal That Different Hydrogen Bonding Patterns Are Possible for G·U Pairs: One Hydrogen Bond for Each G·U Pair in r(GGCGUGCC)₂ and Two for Each G·U Pair in r(GAGUGCUC)₂^†^,^‡

G·U pairs occur frequently and have many important biological functions. The stability of symmetric tandem G·U motifs depends both on the adjacent Watson−Crick base pairs, e.g., 5‘G > 5‘C, and the sequence of the G·U pairs, i.e., 5‘-UG-3‘ > 5‘-GU-3‘, where an underline represents a nucleotide in a G·U pair [Wu, M., McDowell, J. A., and Turner, D. H. (1995) Biochemistry 34, 3204−3211]. In particular, at 37 °C, the motif 5‘-CGUG-3‘ is less stable by approximately 3 kcal/mol compared with other symmetric tandem G·U motifs with G-C as adjacent pairs:  5‘-GGUC-3‘, 5‘-GUGC-3‘, and 5‘-CUGG-3‘. The solution structures of r(GAGUGCUC)2 and r(GGCGUGCC)2 duplexes have been determined by NMR and restrained simulated annealing. The global geometry of both duplexes is close to A-form, with some distortions localized in the tandem G·U pair region. The striking discovery is that in r(GGCGUGCC)2 each G·U pair apparently has only one hydrogen bond instead of the two expected for a canonical wobble pair. In the one-hydrogen-bond model, the distance between GO6 and UH3 is too far to form a hydrogen bond. In addition, the temperature dependence of the imino proton resonances is also consistent with the different number of hydrogen bonds in the G·U pair. To test the NMR models, U or G in various G·U pairs were individually replaced by N3-methyluridine or isoguanosine, respectively, thus eliminating the possibility of hydrogen bonding between GO6 and UH3. The results of thermal melting studies on duplexes with these substitutions support the NMR models