Wen-pi-tang-Hab-Wu-ling-san, a Polyherbal Medicine, Attenuates ER Stress in 3T3-L1 Preadipocytes by Promoting the Insulin Signaling Pathway

The endoplasmic reticulum (ER) is an organelle that functions to synthesize, fold, and transport proteins. ER stress is a key link between type 2 diabetes (T2D), obesity, and insulin resistance. In this study, we investigated the effect of WHW on the ER stress response and the insulin signaling pathway in 3T3-L1 preadipocytes. 3T3-L1 preadipocytes were differentiated into adipocytes, and ER stress was then induced by treatment with tunicamycin. ER stress-induced adipocytes were treated with different concentrations of WHW for 24 h. The expression of ER stress-related molecules such as X-box-binding protein-1 (XBP-1), glucose-regulated protein 78 (GRP78), C/EBP-homologous protein 10 (CHOP10), and eukaryotic initiation factor 2α (eIF2α) and signaling molecules such as phosphatidylinositol 3-kinase (PI3K), insulin receptor substrates-1 (IRS-1), and c-Jun N-terminal protein kinase (JNK) were investigated. WHW significantly inhibited the expression of XBP-1, GRP78, CHOP10, and eIF2α in ER stress-induced 3T3-L1 adipocytes. WHW also increased the PI3K expression and the IRS-1 phosphorylation but decreased the phosphorylation of JNK in ER stress-induced 3T3-L1 adipocytes. Our results indicate that WHW inhibits ER stress in adipocytes by suppressing the expression of ER stress-mediated molecules and the insulin signaling pathway, suggesting that WHW may be an attractive therapeutic agent for managing T2D

Production of α-Bisabolol from metabolically engineered Escherichia coli

α-Bisabolol is a natural-occurring sesquiterpenoid with applications in cosmetics as whitening and soothing agent. It is synthesized from the universal precursors, isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP), which are generated either through the mevalonate (MVA) pathway or the 2C-methyl-D-erythritol-4-phosphate (MEP) pathway. Farnesyl pyrophosphate (FPP) synthase (IspA) then catalyzes the condensation of IPP and DMAPP to the linear FPP, which is rearranged and cyclized to α-bisabolol by bisabolol synthases. Here, we compared the capacity of 5 α-bisabolol synthases from Lippia dulcis, Streptomyces citricolor, Santalum spicatum, Matricaria recutita, and Artemisia annua for α-bisabolol production. MVA pathway and FPP synthase were also overexpressed to supply sufficient FPP for bisabolol synthesis in the recombinant E. coli. Bisabolol synthase from M. recutita (MrBBS) shows the highest activity of bisabolol synthesis, and 75 mg/L/OD600 of bisabolol was produced in a test-tube culture. We further optimized the expression level of IspA and MrBBS by modulation their RBS strength. The 24 bisabolol synthesis operons with different RBSs were assessed for their performance on bisabolol synthesis. By this approach, the best strain is able to produce bisabolol with a capacity of 220mg/L/OD600 in a test tube culture. The consequence of host strain optimization led to an increase in bisabolol production to 300 mg/L/OD600, which presents a 4-fold increase over the initial engineered strain. This work was supported by a grant (NRF-2016R1A2B2010678) from the National Research Foundation, MSIP, Korea

MODULATORY EFFECT OF PERIOSTRACUM CICADAE AND BETULAE CORTEX EXTRACTS ON THE ACTIVATION OF ATOPIC DERMATITIS-RELATED ION CHANNELS ORAI1 AND TRPV3

Background: The cast-off shells of Cryptotympana pustulata (Periostracum Cicadae, PC) and the bark of Betula platyphylla (Betulae Cortex, BC) are used as traditional medicines for the treatment of skin diseases. This study was conducted to investigate the regulatory effects of PC and BC extracts on the activation of the ion channels, calcium release-activated calcium channel protein 1 (ORAI1) and transient receptor potential cation channel subfamily V member 3 (TRPV3). Materials and Methods: Human HEK293T cells, co-overexpressing ORAI1/stromal interaction molecule 1 (STIM1) or overexpressing TRPV3, were treated with PC or BC extracts at 0.1 mg/mL. The changes in ORAI1 and TRPV3 activities were measured using a conventional whole-cell patch-clamp technique. Results: PC and BC extracts significantly decreased ORAI1 activation in ORAI1-STIM1 co-overexpressing HEK293T cells and significantly increased TRPV3 activation in TRPV3 overexpressing cells, compared to that of 2- aminoethoxydiphenyl borate (2-APB, 100 μM), a known agonist of TRPV3. Conclusion: Our results suggest that PC and BC extracts have therapeutic potential to improve skin barrier abnormalities in atopic dermatitis via modulation of ORAI1 and TRPV3 activation

A homozygous Keap1-knockout human embryonic stem cell line generated using CRISPR/Cas9 mediates gene targeting

Kelch-like ECH-associated protein 1 (keap1) is a cysteine-rich protein that interacts with transcription factor Nrf2 in a redox-sensitive manner, leading to the degradation of Nrf2 (Kim et al., 2014a). Disruption of Keap1 results in the induction of Nrf2-related signaling pathways involving the expression of a set of anti-oxidant and anti-inflammatory genes. We generated biallelic mutants of the Keap1 gene using a CRISPR-Cas9 genome editing method in the H9 human embryonic stem cell (hESC). The Keap1 homozygous-knockout H9 cell line retained normal morphology, gene expression, and in vivo differentiation potential. (C) 2016 The Author(s). Published by Elsevier B.V.

Quantitative measurements of C-reactive protein using silicon nanowire arrays

A silicon nanowire-based sensor for biological application showed highly desirable electrical responses to either pH changes or receptor-ligand interactions such as protein disease markers, viruses, and DNA hybridization. Furthermore, because the silicon nanowire can display results in real-time, it may possess superior characteristics for biosensing than those demonstrated in previously studied methods. However, despite its promising potential and advantages, certain process-related limitations of the device, due to its size and material characteristics, need to be addressed. In this article, we suggest possible solutions. We fabricated silicon nanowire using a top-down and low cost micromachining method, and evaluate the sensing of molecules after transfer and surface modifications. Our newly designed method can be used to attach highly ordered nanowires to various substrates, to form a nanowire array device, which needs to follow a series of repetitive steps in conventional fabrication technology based on a vapor-liquid-solid (VLS) method. For evaluation, we demonstrated that our newly fabricated silicon nanowire arrays could detect pH changes as well as streptavidin-biotin binding events. As well as the initial proof-of-principle studies, C-reactive protein binding was measured: electrical signals were changed in a linear fashion with the concentration (1 fM to 1 nM) in PBS containing 1.37 mM of salts. Finally, to address the effects of Debye length, silicon nanowires coupled with antigen proteins underwent electrical signal changes as the salt concentration changed

Comparison of standard versus 90° rotation technique for LMA Flexible™ insertion: a randomized controlled trial

Background Insertion of a flexible laryngeal mask airway (LMA Flexible) is known to be more difficult than that of a conventional laryngeal mask airway. The 90° rotation technique can improve the success rate with a conventional laryngeal mask airway but its effect with the LMA Flexible remains unknown. We assessed whether the 90° rotation technique increased the first-attempt success rate of LMA Flexible insertion versus the standard technique. Methods In total, 129 female patients undergoing breast surgery were analyzed. The primary endpoint was success at the first attempt. The insertion time, number of trials, number of manipulations required, and oropharyngeal leak pressure were also evaluated. Heart rate and mean blood pressure were recorded 1 min before and 1 min after insertion. Blood staining on the LMA Flexible after removal and postoperative sore throat were checked. Results The first-attempt success rates were comparable between the groups (93% vs. 98.3%, P = .20). The insertion time, number of trials and manipulations, hemodynamic variables, and complications, such as blood staining and sore throat, did not differ between the groups. Conclusions The 90° rotation technique is a good alternative to the standard technique for insertion of the LMA Flexible. Trial registration ClinicalTrials.gov(NCT03028896). It was registered retrospectively at Jan 19th, 2017.ASA: American Society of Anesthesiologists; BMI: Body mass index; IV: Intravenous; LMA: Laryngeal mask airway; MAP: Mean arterial pressure; NRS: Numerical rating scale; PACU: Post-anesthetic care unit; SD: Standard deviatio

Barriers to Outpatient Hospital-Based Cardiac Rehabilitation in Korean Patients With Acute Coronary Syndrome

ObjectiveTo investigate factors associated with enrollment and participation in cardiac rehabilitation (CR) in Korea.MethodsPatients admitted to four university hospitals with acute coronary syndrome between June 2014 and May 2016 were enrolled. The Cardiac Rehabilitation Barriers Scale (CRBS) made of 21-item questionnaire and divided in four subdomains was administered during admission. CRBS items used a 5-point Likert scale and ≥2.5 was considered as a barrier. Differences between CR non-attender and CR attender, or CR non-enroller and CR enroller in subscale and each items of CRBS were examined using the chi-square test.ResultsThe CR participation rate in four hospitals was 31% (170 of the 552). Logistical factors (odds ratio [OR]=7.61; 95% confidence interval [CI], 4.62–12.55) and comorbidities/functional status (OR=6.60; 95% CI, 3.95–11.01) were identified as a barrier to CR enrollment in the subdomain analysis. Among patients who were enrolled (agreed to participate in CR during admission), only work/time conflict was a significant barrier to CR participation (OR=2.17; 95% CI, 1.29–3.66).ConclusionDiverse barriers to CR participation were identified in patients with acute coronary syndrome. Providing the tailored model for CR according to the individual patient's barrier could improve the CR utilization. Further multicenter study with large sample size including other CR indication is required

Clinical outcome of patients with refractory Kawasaki disease based on treatment modalities

PurposeAlthough a significant number of reports on new therapeutic options for refractory Kawasaki disease (KD) such as steroid, infliximab, or repeated intravenous immunoglobulin (IVIG) are available, their effectiveness in reducing the prevalence of coronary artery lesions (CAL) remains controversial. This study aimed to define the clinical characteristics of patients with refractory KD and to assess the effects of adjuvant therapy on patient outcomes.MethodsWe performed a retrospective study of 38 refractory KD patients from January 2012 to March 2015. We divided these patients into 2 groups: group 1 received more than 3 IVIG administration+ steroid therapy, (n=7, 18.4%), and group 2 patients were unresponsive to initial IVIG and required steroid therapy or second IVIG (n=31, 81.6%). We compared the clinical manifestations, laboratory results, and echocardiographic findings between the groups and examined the clinical utility of additional therapies in both groups.ResultsA significant difference was found in the total duration of fever between the groups (13.0±4.04 days in group 1 vs. 8.87±2.30 days in group 2; P=0.035). At the end of the follow-up, all cases in group 1 showed suppressed CAL. In group 2, coronary artery aneurysm occurred in 2 patients (6.4 %). All the patients treated with intravenous corticosteroids without additional IVIG developed CALs including coronary artery aneurysms.ConclusionNo statistical difference was found in the development of CAL between the groups. Prospective, randomized, clinical studies are needed to elucidate the effects of adjunctive therapy in refractory KD patients

Effects of Berberine and Hwangryunhaedok-Tang on Oral Bioavailability and Pharmacokinetics of Ciprofloxacin in Rats

Hwangryunhaedok-Tang (HR) and berberine-containing single herbs are used to treat bacterial infection and inflammatory diseases in eastern Asia. The combination of berberine-containing herbal medicines and ciprofloxacin can be an excellent antibacterial chemotherapy against multidrug resistance bacteria. To evaluate the pretreatment effect of berberine and HR, vehicle, berberine (25 and 50 mg/kg/day), and HR (1.4 g/kg/day) were daily administered to rats for five consecutive days. On day 6, ciprofloxacin was administered (10 mg/kg, i.v. and 20 mg/kg, p.o.) to rats. To assess cotreatment effect of berberine and ciprofloxacin, berberine (50 mg/kg) and ciprofloxacin (20 mg/kg) were coadministered by single oral gavage. Pharmacokinetic data were estimated by noncompartmental model. Compared with ciprofloxacin alone (control group), coadministration of berberine (50 mg/kg) and ciprofloxacin significantly decreased Cmax of ciprofloxacin (P<0.05). In addition, the pretreatment of berberine (50 mg/kg/day) and HR (1.4 g/kg/day) significantly decreased Cmax and AUC0→∞, compared with control group (P<0.05). The oral bioavailability of ciprofloxacin was reduced by cotreatment of berberine and pretreatment of berberine and HR. Our results suggest that the expression of P-glycoprotein and organic anion and/or organic cation transporters (OAT/OCT) could take a role in reduced oral bioavailability of ciprofloxacin by berberine and HR