Mutations in the PKM2 exon-10 region are associated with reduced allostery and increased nuclear translocation.

PKM2 is a key metabolic enzyme central to glucose metabolism and energy expenditure. Multiple stimuli regulate PKM2's activity through allosteric modulation and post-translational modifications. Furthermore, PKM2 can partner with KDM8, an oncogenic demethylase and enter the nucleus to serve as a HIF1α co-activator. Yet, the mechanistic basis of the exon-10 region in allosteric regulation and nuclear translocation remains unclear. Here, we determined the crystal structures and kinetic coupling constants of exon-10 tumor-related mutants (H391Y and R399E), showing altered structural plasticity and reduced allostery. Immunoprecipitation analysis revealed increased interaction with KDM8 for H391Y, R399E, and G415R. We also found a higher degree of HIF1α-mediated transactivation activity, particularly in the presence of KDM8. Furthermore, overexpression of PKM2 mutants significantly elevated cell growth and migration. Together, PKM2 exon-10 mutations lead to structure-allostery alterations and increased nuclear functions mediated by KDM8 in breast cancer cells. Targeting the PKM2-KDM8 complex may provide a potential therapeutic intervention

Effects of ranibizumab on human corneal endothelial cells

AbstractPurposeThis study aims to evaluate corneal endothelial changes occurring over a 3-month period after intravitreal injections of ranibizumab in patients with wet age-related macular degeneration.MethodsThis is a prospective case series. A total of 29 patients (29 eyes) received a 0.5-mg intravitreal injection of ranibizumab. Specular microscopy, including measurement of central corneal thickness and endothelial cell count, was performed on each patient prior to and after completing three intravitreal injections.ResultsAll patients received three intravitreal injections and were followed up for a mean of 3 months. There was no significant change in corneal thickness (p = 0.32) or endothelial cell density (p = 0.38) after ranibizumab injections.ConclusionIntravitreal ranibizumab injections (0.5 mg) have no harmful effects on corneal endothelial cells

Monitoring Apnea in the Elderly by an Electromechanical System with a Carbon Nanotube-based Sensor

SummaryBackgroundBreathing, a part of respiration, is one of the vital functions. Breathing disorders are common in the elderly. An effective breathing sensor for real-time detection of apnea is important in clinical critical care. We aimed to construct a real-time warning platform with a combination of carbon nanotubes (CNTs) and related nano-electromechanical system (NEMS) for elderly care.MethodsThrough a specific acid-treated procedure, multiwalled carbon nanotubes (MWCNTs) were immobilized on a thin silicon dioxide (SiO2) film, coated on a heated silicon wafer. Techniques of photolithography and sputtering with chromium and gold were then implemented on the MWCNT film to develop micro-interdigitated electrodes as a base for the breathing sensor. The sensor was equipped with a programmed microchip processor to become a warning detector for abnormal human breathing, namely less than six breaths per minute. Elderly volunteers were enrolled for examining the effective sensitivity of this novel electromechanical device.ResultsThere were 15 elderly volunteers (9 males and 6 females) tested in this experiment. The dynamic analyses of the MWCNT sensor to exhaled breath showed that it had characteristics of rapid response, high aspect ratio, small tip ratio, and high electrical conductivity. Responses of the MWCNT sensor to exhaled breath was recorded according to different performance parameters, i.e., strength, frequency, flow rate, and breath components. In this study, variable pattern-simulated tests showed that a MWCNT sensor combined with a processor could accurately evoke warning signals (100% of sensitivity rate), indicating its effectiveness and usefulness for detecting abnormal breathing rates, especially apnea.ConclusionOur results showed that a new device composed of an NEMS by combining an MWCNT sensor and complementary metal-oxide semiconductor (CMOS) circuits could be integrated to effectively detect apnea in the elderly. This novel device may improve the pattern of safe respiratory care for the elderly in the future

A novel role of sesamol in inhibiting NF-κB-mediated signaling in platelet activation

<p>Abstract</p> <p>Background</p> <p>Platelet activation is relevant to a variety of coronary heart diseases. Our previous studies revealed that sesamol possesses potent antiplatelet activity through increasing cyclic AMP formation. Although platelets are anucleated cells, they also express the transcription factor, NF-κB, that may exert non-genomic functions in platelet activation. Therefore, we further investigated the inhibitory roles of sesamol in NF-κB-mediated platelet function.</p> <p>Methods</p> <p>Platelet aggregation, Fura 2-AM fluorescence, and immunoblotting analysis were used in this study.</p> <p>Results</p> <p>NF-κB signaling events, including IKKβ phosphorylation, IκBα degradation, and p65 phosphorylation, were markedly activated by collagen (1 μg/ml) in washed human platelets, and these signaling events were attenuated by sesamol (2.5~25 μM). Furthermore, SQ22536 and ODQ, inhibitors of adenylate cyclase and guanylate cyclase, respectively, strongly reversed the sesamol (25 μM)-mediated inhibitory effects of IKKβ phosphorylation, IκBα degradation, and p65 phosphorylation stimulated by collagen. The protein kinase A (PKA) inhibitor, H89, also reversed sesamol-mediated inhibition of IκBα degradation. Moreover, BAY11-7082, an NF-κB inhibitor, abolished IκBα degradation, phospholipase C (PLC)γ2 phosphorylation, protein kinase C (PKC) activation, [Ca²⁺]i mobilization, and platelet aggregation stimulated by collagen. Preincubation of platelets with the inhibitors, SQ22536 and H89, both strongly reversed sesamol-mediated inhibition of platelet aggregation and [Ca²⁺]i mobilization.</p> <p>Conclusions</p> <p>Sesamol activates cAMP-PKA signaling, followed by inhibition of the NF-κB-PLC-PKC cascade, thereby leading to inhibition of [Ca²⁺]i mobilization and platelet aggregation. Because platelet activation is not only linked to hemostasis, but also has a relevant role in inflammation and metastasis, our data demonstrating that inhibition of NF-κB interferes with platelet function may have a great impact when these types of drugs are considered for the treatment of cancer and various inflammatory diseases.</p

ATP-Binding Cassette Transporter VcaM from Vibrio cholerae is Dependent on the Outer Membrane Factor Family for Its Function.

Vibrio cholerae ATP-binding cassette transporter VcaM (V. cholerae ABC multidrug resistance pump) has previously been shown to confer resistance to a variety of medically important drugs. In this study, we set to analyse its properties both in vitro in detergent-solubilised state and in vivo to differentiate its dependency on auxiliary proteins for its function. We report the first detailed kinetic parameters of purified VcaM and the rate of phosphate (Pi) production. To determine the possible functional dependencies of VcaM on the tripartite efflux pumps we then utilized different E. coli strains lacking the principal secondary transporter AcrB (Acriflavine resistance protein), as well as cells lacking the outer membrane factor (OMF) TolC (Tolerance to colicins). Consistent with the ATPase function of VcaM we found it to be susceptible to sodium orthovanadate (NaOV), however, we also found a clear dependency of VcaM function on TolC. Inhibitors targeting secondary active transporters had no effects on either VcaM-conferred resistance or Hoechst 33342 accumulation, suggesting that VcaM might be capable of engaging with the TolC-channel without periplasmic mediation by additional transporters. Our findings are indicative of VcaM being capable of a one-step substrate translocation from cytosol to extracellular space utilising the TolC-channel, making it the only multidrug ABC-transporter outside of the MacB-family with demonstrable TolC-dependency

Do female directors will have impact on corporate performance?

The purpose of this study is to explore the influence of female directors on the corporate performance of Chinese banks and Taiwanese financial holding companies. The sample consists of 14 Chinese banks and 15 Taiwanese financial holding companies in 2010–2014. The empirical results show that the proportion of non-executive female directors and independent female directors have significant positive influence on corporate performance. The result shows that the higher the proportion of female directors is, the better off the corporate performance would be. There is robust evidence in the data to conclude that female directors in specific posts can influence a firm’s corporate performance

Discussing the conceptual framework of cultural landscape in Taiwan

This study aims to develop a measureable tool to identify the feature of Taiwan cultural landscape. In-depth interviews from ten scholars were first conducted to design a questionnaire which contained a seven-dimension measurement model of identifying cultural landscape, and then 808 local participants were asked to response this questionnaire for gathering the quantitative data in four cultural significant sites. Exploratory factor analysis and confirmatory factor analysis were applied to confirm the structure of the supposed measurement model from the questionnaire data. The result of factor analysis showed that the confirmed measurement model was less complicated than the supposed measurement model. The cultural feature dimension of cultural landscape was composed of three factors which are environment element, human evidence and traditional custom. And the place meaning dimension of cultural landscape contained cultural atmosphere, life dependence and affective identity. This finding revealed the evidence that local participants’ perception to the feature of cultural landscape was very different from experts’ understanding of the composition of cultural landscape in Taiwan. It also hints the difficulty of conservation for cultural landscape was local people’s disregards to their cultural landscape since these features didn’t have any physical connection with local people’s daily life. This study build a 16-item scale measuring cultural landscape from the perspective of local people in Taiwan, which provide practitioners a reliable and valid analytical tool to assess cultural landscape. And it also offered government agencies a useful foundation to make conservation strategies with local people for cultural landscape

The Vibrio cholerae var regulon encodes a metallo-β-lactamase and an antibiotic efflux pump, which are regulated by VarR, a LysR-type transcription factor

The genome sequence of V. cholerae O1 Biovar Eltor strain N16961 has revealed a putative antibiotic resistance (var) regulon that is predicted to encode a transcriptional activator (VarR), which is divergently transcribed relative to the putative resistance genes for both a metallo-β-lactamase (VarG) and an antibiotic efflux-pump (VarABCDEF). We sought to test whether these genes could confer antibiotic resistance and are organised as a regulon under the control of VarR. VarG was overexpressed and purified and shown to have β-lactamase activity against penicillins, cephalosporins and carbapenems, having the highest activity against meropenem. The expression of VarABCDEF in the Escherichia coli (ΔacrAB) strain KAM3 conferred resistance to a range of drugs, but most significant resistance was to the macrolide spiramycin. A gel-shift analysis was used to determine if VarR bound to the promoter regions of the resistance genes. Consistent with the regulation of these resistance genes, VarR binds to three distinct intergenic regions, varRG, varGA and varBC located upstream and adjacent to varG, varA and varC, respectively. VarR can act as a repressor at the varRG promoter region; whilst this repression was relieved upon addition of β-lactams, these did not dissociate the VarR/varRG-DNA complex, indicating that the de-repression of varR by β-lactams is indirect. Considering that the genomic arrangement of VarR-VarG is strikingly similar to that of AmpR-AmpC system, it is possible that V. cholerae has evolved a system for resistance to the newer β-lactams that would prove more beneficial to the bacterium in light of current selective pressures