In the title compound, C19H25Cl2NOSi, the NH group and the carbonyl O atom of the amide fragment are involved in an intermolecular N—H⋯O hydrogen bond forming chains of molecules. The plane of the benzene ring forms a dihedral angle of 50.5 (2)° with respect to the silolane ring and an angle of 49.74 (2)° with the cyclopentyl moiety.
Gliomas are the leading cause of death among adults with primary brain malignancies. Treatment for malignant gliomas remains limited, and targeted therapies have been incompletely explored. In this study, we found that the protein expression of presenilin 2 (PS2) was significantly increased in glioma tissues, at least partially because of promoter demethylation. We further evaluated the biological functions of PS2 in U251 glioma cell proliferation, migration, invasion, and tumor growth in vivo by specific inhibition of PS2 using short hairpin RNA (shRNA). We found that PS2 depletion inhibited glioma cell growth as the result of inhibited proliferation and induced apoptosis. PS2 depletion also decreased the invasive capability of glioma cells and anchorage-independent colony formation in soft agar. Moreover, suppression of PS2 expression significantly impaired the growth of glioma xenografts in nude mice. Finally, the decrease in glioma cell growth caused by PS2 depletion seems to involve Nrg1/ErbB signaling. In summary, our data highlight the use of RNA interference (RNAi) as a tool to better understand the molecular basis of PS2 in glioma progression and to uncover new targets for the treatment of glioma.
glioma; invasion; Nrg1/ErbB; presenilin 2; proliferation; RNAi
Different cell/tissue derived extracellular matrix (ECM) display subtle differences that might provide important cues for proliferation and differentiation of cells in vitro or in vivo. However, the bioactivities of different ECMs in vitro were not fully understood. In this study, osteoblasts-derived and fibroblast-derived ECM-coated cell culture dishes were prepared respectively by culturing osteoblastic MC3T3-E1 cells and rat fibroblast then decellularizing the cultures. We investigated the bioactivities of the two different ECMs coated on cell culture plates using cellular, biochemical and molecular method. The proliferative activity of the bone marrow-derived mesenchymal stem cells (BMSCs) cultured on osteoblast-ECM was lower than for BMSCs grown on fibroblast-ECM. Compared with the BMSCs cultured on fibroblast-derived ECM, the cells grown on osteoblastic ECM showed enhanced alkaline phosphatase (ALP) activity, higher BMP-2 and osteopontin protein levels, increased secreted calcium content, and higher levels of runt-related transcriptional factor 2 (Runx 2) and osteocalcin (OCN) mRNA. Knockdown of BMP-2 or FGF-2 with shRNA transfection hardly effected osteoblastic differentiation or proliferation of MC3T3-E1 seeded on osteoblast-ECM or fibroblast-ECM. Therefore, the osteoblastic ECM had better osteoinductive potential and lower proliferative effect than fibroblastic ECM, and the two ECM presented enough bioactivity, knockdown of growth factors had no significant effect on differentiation and proliferation of re-seeded cells.
Extracellular matrix; Bone marrow-derived mesenchymal stem cells; Osteoblasts; siRNA; Proliferation; Differentiation
Transcription factors of the FoxO (forkhead box O) family regulate a wide range of cellular physiological processes, including metabolic adaptation and myogenic differentiation. The transcriptional activity of most FoxO members is inhibitory to myogenic differentiation and overexpression of FoxO1 inhibits the development of oxidative type I fibres in vivo. In this study, we found that FoxO6, the last discovered FoxO family member, is expressed ubiquitously in various tissues but with higher expression levels in oxidative tissues, such as brain and oxidative muscles. Both the expression level and promoter activity of FoxO6 were found to be enhanced by PGC-1α (peroxisome-proliferator-activated receptor γ co-activator 1α), thus explained its enriched expression in oxidative tissues. We further demonstrated that FoxO6 represses the expression of PGC-1α via direct binding to an upstream A/T-rich element (AAGATATCAAAACA,−2228–2215) in the PGC-1α promoter. Oxidative low-intensity exercise induced PGC-1α but reduced FoxO6 expression levels in hind leg muscles, and the binding of FoxO6 to PGC-1α promoter was also prevented by exercise. As FoxO6 promoter can be co-activated by PGC-1α and its promoter in turn can be repressed by FoxO6, it suggests that FoxO6 and PGC-1α form a regulatory loop for setting oxidative metabolism level in the skeletal muscle, which can be entrained by exercise.
FoxO6; muscle; oxidative metabolism; PGC-1α; promoter; transcription; CHIP, chromatin immunoprecipitation; DM, differentiation medium; DMEM, Dulbecco’s modified Eagle’s medium; EMSA, electrophoretic mobility-shift assay; FCS, fetal calf serum; FoxO, forkhead box O; Gapdh, glyceraldehydes-3-phosphate dehydrogenase; GC, gastrocnemius; GM, growth medium; HEK-293T, HEK-293 cells expressing the large T-antigen of SV40; HRP, horseradish peroxidase; MBP, maltose-binding protein; MRFs, myogenic regulatory factors; PBST, PBS containing 0.5% Tween 20; PGC-1α, peroxisome-proliferator-activated receptor γ co-activator 1α; PI3k, phosphoinositide 3-kinase; qRT-PCR, quantitative reverse transcription PCR; T2D, type 2 diabetes
Objectives. We report the single-incision laparoscopic cholecystectomy (SILC) learning experience of 2 hepatobiliary surgeons and the factors that could influence the learning curve of SILC. Methods. Patients who underwent SILC by Surgeons A and B were studied retrospectively. Operating time, conversion rate, reason for conversion, identity of first assistants, and their experience with previous laparoscopic cholecystectomy (LC) were analysed. CUSUM analysis is used to identify learning curve. Results. Hundred and nineteen SILC cases were performed by Surgeons A and B, respectively. Eight cases required additional port. In CUSUM analysis, most conversion occurred during the first 19 cases. Operating time was significantly lower (62.5 versus 90.6 min, P = 0.04) after the learning curve has been overcome. Operating time decreases as the experience increases, especially Surgeon B. Most conversions are due to adhesion at Calot's triangle. Acute cholecystitis, patients' BMI, and previous surgery do not seem to influence conversion rate. Mean operating times of cases assisted by first assistant with and without LC experience were 48 and 74 minutes, respectively (P = 0.004). Conclusion. Nineteen cases are needed to overcome the learning curve of SILC. Team work, assistant with CLC experience, and appropriate equipment and technique are the important factors in performing SILC.
Accumulating evidence shows that enhancer of zeste homolog 2 (E2H2) is upregulated in a broad range of cancer types, such as breast cancer, prostate cancer, ovarian cancer, and colon cancer. Therefore, inhibiting EZH2 expression may be a promising strategy for anticancer therapy. This review focuses on the current understanding of the mechanisms underlying EZH2 regulation that are involved in cancer progression. Also, it introduces two EZH2 inhibitors that target EZH2 and could be potentially applied in the treatment of cancer in the future.
EZH2; PRC2; cancer
In human cancer, expression of the let-7 family is significantly reduced, and this is associated with shorter survival times in patients. However, the mechanisms leading to let-7 downregulation in cancer are still largely unclear. Since an alteration in copy-number is one of the causes of gene deregulation in cancer, we examined copy number alterations of the let-7 family in 2,969 cancer specimens from a high-resolution SNP array dataset. We found that there was a reduction in the copy number of let-7 genes in a cancer-type specific manner. Importantly, focal deletion of four let-7 family members was found in three cancer types: medulloblastoma (let-7a-2 and let-7e), breast cancer (let-7a-2), and ovarian cancer (let-7a-3/let-7b). For example, the genomic locus harboring let-7a-3/let-7b was deleted in 44% of the specimens from ovarian cancer patients. We also found a positive correlation between the copy number of let-7b and mature let-7b expression in ovarian cancer. Finally, we showed that restoration of let-7b expression dramatically reduced ovarian tumor growth in vitro and in vivo. Our results indicate that copy number deletion is an important mechanism leading to the downregulation of expression of specific let-7 family members in medulloblastoma, breast, and ovarian cancers. Restoration of let-7 expression in tumor cells could provide a novel therapeutic strategy for the treatment of cancer.
To estimate the cost and patterns of expenditure of dry eye treatment.
We retrieved data on the type and cost of dry eye treatment in Singapore National Eye Centre from pharmacy and clinic inventory databases over a 2 year period (2008–2009) retrospectively. According to the type of treatment, data were sorted into 7 groups; meibomien gland disease (MGD) treatment, preservative free lubricant eye drops, preserved lubricant eye drops, lubricant ointments and gels, cyclosporine eye drops, oral supplements and non-pharmacological treatments/procedures. Each recorded entry was considered as one patient episode (PE). Comparisons in each group between two years were carried out using Pearson Chi-Square test. Significance level was set at alpha = 0.05.
Cost data from 54,052 patients were available for analysis. Total number of recorded PEs was 132,758. Total annual expenditure on dry eye treatment for year 2008 and 2009 were US$1,509,372.20 and US$1,520,797.80 respectively. Total expenditure per PE in year 2008 and 2009 were US$22.11 and US$23.59 respectively. From 2008 to 2009, there was a 0.8% increase in total annual expenditure and 6.69% increase in expenditure per PE. Pharmacological treatment attributes to 99.2% of the total expenditure with lubricants accounting for 79.3% of the total pharmacological treatment expenditure. Total number of units purchased in preservative free lubricants, cyclosporine eye drops and MGD therapy have increased significantly (p<0.001) whereas number of units purchased in preserved lubricants and ointments/gels have reduced significantly (p<0.001) from 2008 to 2009.
Dry Eye imposes a significant direct burden to health care expenditure even without considering indirect costs. Health care planners should be aware that these direct costs appear to increase over the time and more so for particular types of medications. Given the limitations of socio-economic data, true societal costs of Dry eye syndrome are likely to be much higher than estimated.
Background. Alzheimer's disease (AD) is the most common cause of dementia in the elderly population. Growing evidence supports that AD patients are at high risk for hip fracture, but the issue remains questionable. The purpose of the present study is to perform a meta-analysis to explore the association between AD and risk of hip fracture. Considering that bone mineral density (BMD) acts as a strong predictor of bone fracture, we also studied the hip BMD in AD patients. Methods. We searched all publications in Medline, SciVerse Scopus, and Cochrane Library published up to January 2012 about the association between AD and hip fracture or hip BMD. Results. There are 9 studies included in the meta-analysis. The results indicate that AD patients are at higher risk for hip fracture (OR and 95% CI fixed: ES = 2.58, 95% CI = [2.03, 3.14]; dichotomous data: summary OR = 1.80, 95% CI = [1.54, 2.11]) than healthy controls. Further meta-analysis showed that AD patients have a lower hip BMD (summary SMD = −1.12, 95% CI = [−1.34, −0.90]) than healthy controls. Conclusions. It was found that in comparison with healthy controls AD patients are at higher risk for hip fracture and have lower hip BMD.
Background. Many studies have reported an association between tea drinking and Parkinson's disease (PD). Our purpose is to summarize the available information and evaluate the risk of PD associated with tea drinking. Methods. We searched all publications in English language on the association of tea drinking and PD risk published up to December 2010. The pooled analysis was performed with Review Manager 5.0. Results. In total, eight articles including 1418 cases and 4250 controls were included in the meta-analysis. The pooled odds ratio (95% CI) was 0.85 (0.74–0.98), which suggests the protective effect of tea drinking in PD risks. Moreover, the summary OR (OR: 0.83, 95% CI = 0.69–0.99) for drinkers of ≤1 cup of tea per day versus nonconsumers and that (OR: 0.96, 95% CI = 0.73–1.27) for drinkers of >1 cups of tea per day versus nonconsumers showed that there was not an apparent dose-response relationship. No indication for publication bias was found. Conclusions. This meta-analysis showed that tea drinking can lower the risk of PD, while no apparent dose-response relationship was found. Further effort is needed to fully understand the mechanism underlying the beneficial effect of tea consumption in lowering PD risk.
In the title homochiral mononuclear compound, [Zn(NO3)2(C6H12N4O2)2], the ZnII atom is located on a twofold rotation axis and coordinated by two N atoms from two ligands and two O atoms from two NO3
− anions, adopting a distorted tetrahedral coordination geometry. The compound is enantiomerically pure and corresponds to the S diastereoisomer, with the optical activity originating from the chiral ligand. In the crystal, molecules are connected into three-dimensional supramolecular networks through O—H⋯O, O—H⋯N and N—H⋯O hydrogen bonds.
Altered metabolism is one of the most significant features of cancer cells. ATP citrate lyase (ACL), a key enzyme in de novo lipid synthesis, has been reported to be overexpressed or activated in several cancer types. To determine the role of ACL in ovarian cancer progression, we detected ACL expression in human epithelial ovarian cancer tissues. qRT-PCR and western blotting showed higher ACL expression in malignant tissues compared to normal ovarian tissues. Immunohistochemical analysis showed that phosphorylated ACL was increased in ovarian cancer tissues and that its expression correlated well with tumor grade, FIGO stage and poorer prognosis. To explore the therapeutic potential of ACL, we assessed the effect of ACL-siRNA on cellular proliferation and cell cycle distribution. ACL knockdown inhibited cellular proliferation and induced cell cycle arrest in A2780 cells. Taken together, our findings suggest that ACL may contribute to the pathogenesis of human epithelial ovarian cancer, and may serve as a novel therapeutic target.
ATP citrate lyase; ovarian cancer; prognosis; SREBP-1; RNAi
The natural isoquinoline alkaloid berberine possesses potential to treat Alzheimer's disease (AD) by targeting multiple pathogenic factors. In the present study, docking simulations were performed to gain deeper insights into the molecular basis of berberine's inhibitory effects against the important pathogenic enzymes of AD, that is, acetylcholinesterase, butyrylcholinesterase, and two isoforms of monoamine oxidase. It was found that the theoretical binding affinities of berberine to the four enzymes are very close to the experimental values, which verify the methodology. Further inspection to the binding modes found that hydrophobic interactions between the hydrophobic surface of berberine and neighboring hydrophobic residues are the principal forces contributing to the ligand-receptor interactions. Although berberine cation also has potential to form electrostatic interaction with neighboring residues, it is interesting to find that electrostatic force is excluded in the four cases unexpectedly. These results have important implications for the berberine-based anti-AD drug design.
It is now widely accepted that at an early stage in the evolution of life an RNA world arose, in which RNAs both served as the genetic material and catalyzed diverse biochemical reactions. Then, proteins have gradually replaced RNAs because of their superior catalytic properties in catalysis over time. Therefore, it is important to investigate how primitive functional proteins emerged from RNA world, which can shed light on the evolutionary pathway of life from RNA world to the modern world. In this work, we proposed that the emergence of most primitive functional proteins are assisted by the early primitive nucleotide cofactors, while only a minority are induced directly by RNAs based on the analysis of RNA-protein complexes. Furthermore, the present findings have significant implication for exploring the composition of primitive RNA, i.e., adenine base as principal building blocks.
In the title compound, C20H26N2O4S2, the cyclohexane ring has a chair conformation. The two chiral C atoms are in S configurations. In the crystal, intermolecular N—H⋯O hydrogen bonds link the molecules into chains propagating in . Weak intermolecular C—H⋯O hydrogen bonds further stabilize the crystal packing.
We determined the proportion of foreign-born persons with tuberculosis (TB) in Singapore. This proportion increased from 25.5% in 2004 to 37.6% in 2009. Unskilled workers from countries with high incidences of TB accounted for the highest number of and greatest increase in foreign-born TB case-patients.
Epidemiology; tuberculosis; tuberculosis and other mycobacteria; bacteria; foreign-born persons; Singapore; dispatch
The runt-related transcription factor, Runx2 may have an oncogenic role in mediating metastatic events in breast cancer, but whether Runx2 has a role in the early phases of breast cancer development is not clear. We examined the expression of Runx2 and its relationship with estrogen receptor (ER) and progesterone receptor (PR) in breast cancer cell lines and tissues.
Two human breast cancer cell lines, MCF-7 and MDA-MB-231 were transiently transfected with vectors expressing either Runx2 or ER and the levels of both proteins and mRNA were examined by Western blot analysis and quantitative real-time PCR respectively. Runx2 expression was also examined in tissue microarray sections of 123 breast cancer patients by immunohistochemistry and results were correlated with clinico-pathological characteristics.
Expression of Runx2 and ER was reciprocal in the breast cell culture models and Runx2 suppressed ERβ but ERα not mRNA levels. In contrast, functional expression of Runx2 was evident in the nucleus in 28% of the breast cancer tissues and in both early and late stages of tumor growth. Importantly, Runx2 expression was significantly more frequent in Grade 2 compared to Grade 1 and Grade 3 tumors (48% vs 39% vs 13%) and the expression was significantly associated with ER (p=0.005), PR (p=0.008) expression in Grade 2 & 3 tumors than the Grade 1 tumors.
We propose that Runx2, ER and PR triple positivity in Grade 2 & 3 defines a biological subtype in breast cancer.
Runx2; breast cancer; estrogen receptor; progesterone receptor; CerbB2
Two-dimensional echocardiographic (2DE) continuity-equation derived aortic valve area (AVA) in aortic stenosis (AS) relies on non-simultaneous measurement of left ventricular outflow tract (LVOT) velocity and geometric assumptions of LVOT area, which can amplify error, especially in upper septal hypertrophy (USH). We hypothesized that real-time three-dimensional echocardiography (RT3DE) can improve accuracy of AVA by directly measuring LVOT stroke volume (SV) in one window.
Methods and results
RT3DE colour Doppler and 2DE were acquired in 68 AS patients (74 ± 12 yrs) prospectively. SV was derived from flow obtained from a sampling curve placed orthogonal to LVOT (Tomtec Imaging). Agreement between continuity-equation derived AVA by RT3DE (AVA3D-SV) and 2DE (AVA2D) and predictors of discrepancies were analysed. Validation of LVOT SV was performed by aortic flow probe in a sheep model with balloon inflation of septum to mimic USH. There was only modest correlation between AVA2D and AVA3D-SV (r = 0.71, difference 0.11 ± 0.23 cm2). The degree of USH was significantly associated with difference in AVA calculation (r = 0.4, P = 0.005). In experimentally distorted LVOT geometry in sheep, RT3DE correlated better with flow probe assessment (r = 0.96, P < 0.001) than 2DE (r = 0.71, P = 0.006).
RT3DE colour Doppler-derived LVOT SV in the calculation of AVA by continuity equation is more accurate than 2D, including in situations such as USH, common in the elderly, which modify LVOT geometry.
Aortic stenosis; Real-time three-dimensional echocardiography; Colour Doppler; Valvular heart disease; Continuity equation
In the present work, a theoretical study on the deactivation of triplet excited (T1) state thioxanthone (TX) by indole (INH) was performed, based on density functional theory calculations. Three feasible pathways, namely direct electron transfer from INH to T1 state TX, electron transfer followed by proton transfer from INH.+ to TX.−, and H-atom transfer from nitrogen of INH to keto oxygen of T1 state TX, were proposed theoretically to be involved in T1 state TX deactivation by INH.
thioxanthone; indole; triplet excited state; deactivation; quantum chemical calculation
In the title compound, C8H12N2·H2O, four substituted pyridine molecules alternate with four water molecules, forming a large ring via Owater—H⋯Npyridine and Namine—H⋯Owater hydrogen bonding. Adjacent rings are connected via Owater—H⋯Owater hydrogen-bonds, forming a three-dimensional network.
The deactivation of triplet excited state riboflavin by polyphenols, e.g. rutin and catechin, was studied on the basis of density functional theory calculations. The results show that the H-atom transfer pathway is more feasible on thermodynamic grounds in comparison with the direct energy transfer or direct electron transfer pathways involved in the triplet excited state riboflavin deactivation by rutin/catechin. The findings are helpful to understand the protective effect of polyphenols against the riboflavin induced photosensitizing damage.
Riboflavin; polyphenols; triplet excited state; deactivation; density functional theory
Ligand-protein mapping was found to follow a power law and the preferential attachment principle, leading to the identification of the molecules, mostly nucleotide-containing compounds, that are likely to have evolved earliest.
Extant life depends greatly on the binding of small molecules (such as ligands) with macromolecules (such as proteins), and one ligand can bind multiple proteins. However, little is known about the global patterns of ligand-protein mapping.
By examining 2,186 well-defined small-molecule ligands and thousands of protein domains derived from a database of druggable binding sites, we show that a few ligands bind tens of protein domains or folds, whereas most ligands bind only one, which indicates that ligand-protein mapping follows a power law. Through assigning the protein-binding orders (early or late) for bio-ligands, we demonstrate that the preferential attachment principle still holds for the power-law relation between ligands and proteins. We also found that polar molecular surface area, H-bond acceptor counts, H-bond donor counts and partition coefficient are potential factors to discriminate ligands from ordinary molecules and to differentiate super ligands (shared by three or more folds) from others.
These findings have significant implications for evolution and drug discovery. First, the chronology of ligand-protein binding can be inferred by the power-law feature of ligand-protein mapping. Some nucleotide-containing ligands, such as ATP, ADP, GDP, NAD, FAD, dihydro-nicotinamide-adenine-dinucleotide phosphate (NDP), nicotinamide-adenine-dinucleotide phosphate (NAP), flavin mononucleotide (FMN) and AMP, are found to be the earliest cofactors bound to proteins, agreeing with the current understanding of evolutionary history. Second, the finding that about 30% of ligands are shared by two or more domains will help with drug discovery, such as in finding new functions from old drugs, developing promiscuous drugs and depending more on natural products.