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Methylthioadenosine phosphorylase (MTAP) is a rate-limiting enzyme in the methionine salvage pathway, which recycles one carbon unit that is lost during polyamine synthesis back into the methionine cycle. Although MTAP deficiency has been reported in various tumours, MTAP is overexpressed and might promote oncogenesis in other cancers, including prostate and colon cancer. Currently, little is known about the MTAP status of oral squamous cell carcinoma (OSCC). In this study, we immunohistochemically examined the expression of MTAP in surgically resected oral epithelial dysplasia (OED, n=7), carcinoma in situ (CIS) (n=16), and OSCC (n=118). In the normal epithelium, MTAP was only weakly expressed in the cytoplasm of the basal layer cells. In OED, CIS, and OSCC, MTAP was uniformly expressed in the cytoplasm of the dysplastic and cancer cells. In addition to cytoplasmic MTAP expression, 45 of 118 cases (38.1%) exhibited increased nuclear expression of MTAP in the cancer cells at the invasive front. Statistical analysis showed that the concomitant nuclear and cytoplasmic expression of MTAP was associated with a high budding score (p=0.0023); poor differentiation (p=0.0044); aggressive invasion patterns (p=0.0001); and features of epithelial-to-mesenchymal transition (EMT), such as loss of E-cadherin expression (p=0.0003) and upregulated expression of vimentin (p=0.0002), slug (p=0.0002), and laminin 5 (p

In this article, current understandings of neurotransmitters and neuropharmacology of sleep/wake regulations are discussed. Sleep/wake is a complex physiology regulated by brain activities in multiple brain sites, and multiple neurotransmitter systems such as monoamines, acetylcholine, excitatory and inhibitory amino acids, peptides, purines, and neuronal and nonneuronal humoral modulators are likely to be involved. With recent advances in neuroscience research, neurobiology of wakefulness, nonrapid eye movement (NREM) sleep, and rapid eye movement (REM) sleep pathways have been refined from the classical models of sleep/wake pathways. Sleep disturbances can be induced intrinsically and extrinsically, and the functional disturbance of these neurotransmitter systems are likely to cause sleep disturbances. Some of these neurotransmitters have been recognized as target molecules for the treatment of common sleep disorders, such as γ-aminobutyric acid (GABA) enhancers as hypnotics and presynaptic dopamine enhancers as wake-promoting compounds. Gaining the knowledge of neurobiology of sleep/wake is essential for developments of better pharmacological treatments for sleep disorders.

An improved process for preparing tenuifolin (presenegenin 3-β-d-glucopyranoside) from the root of Polygala senega L. was developed. A crude saponin mixture extracted from P. senega was subjected to hydrolysis, and the reactivity of compounds in the extract was controlled by utilizing the combination of a flow reactor and experimental design. In addition, column chromatography with HP 20, a synthetic polystyrenic adsorbent, allowed the gram-scale preparation of tenuifolin in a continuous manner with fewer steps. This approach shortens the total time required for gram-scale preparation from 16 to 5 h in a continuous manner while improving the yield from 0.59% to 2.08% (w/w).

Thermal transport properties of patterned binary self-assembled monolayers (SAMs) on Au(111) were examined using scanning thermal microscopy (SThM) with both contact and noncontact methods. We fabricated two-dimensional (2D) patterns with two separate domains of n-hexadecanethiol/benzenethiol, benzenethiol/n-butanethiol, or n-hexadecanethiol/n-butanethiol. In the experimental setup, the efficiency of thermal transport from a SThM tip to the SAM surface can be evaluated in terms of the temperature change at the SThM tip. In the contact regime, where a SThM tip physically contacts the SAM surface, direct thermal transport through the SAM and radiation-based thermal transport through the space where SAMs exist may contribute to a drop in temperature at the tip. In the noncontact regime, thermal transport relies on radiation-based heat dissipation from the heated tip to the SAMs. 2D mapping of the spatial temperature distribution on SAMs reflects the difference in thermal transport properties of the two SAM domains. We found that the contact method is effective for visualizing the temperature contrast, which reflects the thermal transport properties of the constituent molecules when the domains of the SAMs have a similar height, while the noncontact method allows visualization of the temperature distribution, which is related to the height of each domain of the SAMs, rather than the chemical structures of the constituent molecules. Combination of contact and noncontact SThM enables 2D imaging of thermal transport properties and topographic imaging simultaneously and represents a new technique for investigating the thermal properties of materials surfaces, which is essential for nanoscale thermal management.

Myostatin (Mstn) and GDF11 are critical factors that are involved in muscle atrophy in the young and sarcopenia in the elderly, respectively. These TGF-β superfamily proteins activate not only Smad signalling but also non-Smad signalling including the Ras-mediated ERK pathway (Raf–MEK–ERK phosphorylation cascade). Although Mstn and GDF11 have been shown to induce muscle atrophy or sarcopenia by Smad2/3-mediated Akt inhibition, participation of the non-Smad Ras–ERK pathway in atrophy and sarcopenia has not been well determined. We show here that both Mstn and GDF11 prevented skeletal myocyte differentiation but that the MEK inhibitor U0126 or trametinib restored differentiation in Mstn- or GDF11-treated myocytes. These MEK inhibitors induced the expression of DA-Raf1 (DA-Raf), which is a dominant-negative antagonist of the Ras–ERK pathway. Exogenous expression of DA-Raf in Mstn- or GDF11-treated myocytes restored differentiation. Furthermore, administration of trametinib to aged mice resulted in an increase in myofiber size or recovery from muscle atrophy. The trametinib administration downregulated ERK activity in these muscles. These results imply that the Mstn/GDF11-induced Ras–ERK pathway plays critical roles in the inhibition of myocyte differentiation and muscle regeneration, which leads to muscle atrophy. Trametinib and similar approved drugs might be applicable to the treatment of muscle atrophy in sarcopenia or cachexia.

Arginine vasopressin (AVP) is a hormone that is mainly synthesized in the hypothalamus and stored in the posterior pituitary. Receptors for vasopressin are categorized into at least three subtypes (V1A, V1B, V2). Among these subtypes, the V1B receptor (V1BR), highly expressed in the pituitary, is a primary regulator of the hypothalamic-pituitary-adrenal axis activity, and thus a potential target for the treatment of neuropsychiatric disorders, such as depression and anxiety. 11C-TASP699 is a novel PET radiotracer with high affinity and selectivity for the V1BR. The purpose of this study was to characterize the pharmacokinetic and binding profiles of 11C-TASP699 in human and determine its utility in an occupancy study of a novel V1BR antagonist, TS-121. Methods: Six healthy subjects were scanned twice with 11C-TASP699 to determine the most appropriate kinetic model for analysis of imaging data and test-retest reproducibility of outcome measures. Nine healthy subjects were scanned before and after administration of TS-121 (active component: THY1773) to assess V1BR occupancy. Metabolite-corrected arterial input functions were obtained. Pituitary time-activity curves were analyzed with one- and two-tissue compartment (1TC, 2TC) models and multilinear analysis 1 (MA1) to calculate distribution volumes (V T). Relative test-retest variability (TRV) and absolute test-retest variability (aTRV) were calculated. Since no brain region could be used as a reference region, percent change in V T after TS-121 administration was computed to assess its receptor occupancy and correlate with plasma concentration of the drug. Results: 11C-TASP699 showed high uptake in the pituitary and no uptake in any brain regions. The 2TC model provided better fits than the 1TC model. The MA1 V T estimates were very similar to the 2TC V T estimates, so MA1 was the model of choice. TRV of V T was good (TRV: -2 ± 14%, aTRV: 11%). THY1773 reduced VT in a dose-dependent fashion, with IC50 of 177 ± 52 ng/mL in plasma concentration. There were no adverse events resulting in discontinuation from the study. Conclusion: 11C-TASP699 was shown to display appropriate kinetics in human with substantial specific binding and good reproducibility of V T Therefore, this tracer is suitable for measurement of the V1BR in human pituitary and V1BR occupancy of TS-121, a novel V1BR antagonist.

The interphase structure and properties of isotactic polypropylene (it.PP) have received much attention in the fundamental science and industrial applications for the improvement of the poor adhesion properties of the it.PP. In this work, we focused on the laminates with it.PP substrates and various ethylene–octene rubber (EOR) hot-melt adhesives with different molecular weights and crystallinities. We prepared laminate samples using EOR with low molecular weight, random copolymerized EOR, and block-like copolymerized EOR as adhesives. We performed T-peel tests of the laminate samples and evaluated their thickness of the interphase with confocal Raman scattering imaging, X-ray refraction contrast imaging, and atomic force microscopic imaging. The laminates with larger interphase thicknesses possessed the larger T-peel strengths, which were relatively consistent with these obtained results among all the measurement methods. It is revealed that the adhesion properties and the interphase thicknesses depended on the crystallinities of the it.PP substrates. These results mean that the interphase would be constructed by not only diffusion of molecular chains but also the crystalline growth and the formation of lamellae interlock in the interphase region.

Background It is difficult to distinguish between a tumor and its liver segment with traditional use of indocyanine green (ICG) alone. In the present study, a method was used to limit ICG to the liver segment adjacent to a tumor. A spectrally-distinct fluorescently-labeled tumor-specific antibody against human carcinoembryonic antigen-related cell-adhesion molecules was used to label the metastatic tumor in a patient-derived orthotopic xenograft mouse model to enable color-coded visualization and distinction of a colon-cancer liver metastases and its adjacent liver segment. Materials and Methods Nude mice received surgical orthotopic implantation in the liver of colon-cancer liver metastases derived from two patients. An anti- carcinoembryonic antigen-related cell-adhesion molecules monoclonal antibody (mAb 6G5j) was conjugated to a near-infrared dye IR700DX (6G5j-IR700DX). After three weeks, mice received 6G5j-IR700DX via tail-vein injection 48 hours before surgery. ICG was intravenously injected after ligation of the left or left lateral Glissonean pedicle resulting in labeling of the segment with preserved blood-flow in the liver. Imaging was performed with the Pearl Trilogy and FLARE Imaging Systems. Results The metastatic liver tumor had a clear fluorescence signal due to selective tumor targeting by 6G5j-IR700DX, which was imaged on the 700 nm channel. The adjacent liver segment, with preserved blood-flow in the liver, had a clear fluorescence ICG 800 nm signal, while the left or left lateral segment had no fluorescence signal. Overlay of the images showed clear color-coded differentiation between the tumor fluorescing at 700 nm and the adjacent liver segment fluorescing at 800 nm. Conclusions Color-coding of a liver tumor and uninvolved liver segment has the potential for improved liver resection.

The random copolymer of chloroprene and acrylonitrile is a newly developed rubber whose features and value propositions are not scientifically explored yet. This article focuses on the basic characterizations and properties of acrylonitrile-chloroprene rubber. Qualitative analyses through infrared (FTIR) and nuclear magnetic resonance (1H-NMR) spectra confirm the presence of both the -Cl and -CN groups in the new rubber. As evidenced through differential scanning calorimetry (DSC) and dynamic mechanical thermal analysis (DMTA), the single glass transition temperature of acrylonitrile-chloroprene rubber reflects its monophasic random microstructure. While compared against commercial grades of chloroprene rubber (CR) and nitrile rubber (NBR), the new rubber provides a distinctive combination of properties that are not available with either of the elastomer alone. Acrylonitrile-chloroprene rubber demonstrates slightly lower specific gravity, an improved low-temperature compression set, higher flex-fatigue resistance, and lower volume swelling in IRM 903 and Fuel C to chloroprene rubber. As compared to nitrile rubber, the new copolymer shows appreciably better heat aging and ozone resistance. Good abrasion resistance, low heat buildup, and remarkably high flex-fatigue resistance indicate excellent durability of the acrylonitrile-chloroprene rubber under dynamic loading. Based on the preliminary results, it is apparent that the new copolymer can be a candidate elastomer for various industrial applications which demand good fluid resistance, high heat and low-temperature tolerances, good weatherability, and durability under static and dynamic conditions.

A series of six title compounds have been prepared using 3-amino- 6-chloropyridazine as starting material collected from commercial sources and were characterized by IR, 1H NMR, and high-resolution mass spectral (HRMS) data. The method involves three steps: Suzuki-Miyaura cross-coupling reaction, N(2)-alkylation, and acid hydrolysis, respectively, to obtain final products with good yields. According to Lipinski's rule of five and Veber's rule, pharmacokinetics studies of the synthesized compounds showed that all the parameters were in between the permissible limits. The toxicity parameters were low for the compounds to act as drugs. J. Bangladesh Acad. Sci. 45(1); 37-47: June 2021

Mutations in the type 1 ryanodine receptor (RyR1), a Ca2+ release channel in skeletal muscle, hyperactivate the channel to cause malignant hyperthermia (MH) and are implicated in severe heat stroke. Dantrolene, the only approved drug for MH, has the disadvantages of having very poor water solubility and long plasma half-life. We show here that an oxolinic acid-derivative RyR1-selective inhibitor, 6,7-(methylenedioxy)-1-octyl-4-quinolone-3-carboxylic acid (Compound 1, Cpd1), effectively prevents and treats MH and heat stroke in several mouse models relevant to MH. Cpd1 reduces resting intracellular Ca2+, inhibits halothane- and isoflurane-induced Ca2+ release, suppresses caffeine-induced contracture in skeletal muscle, reduces sarcolemmal cation influx, and prevents or reverses the fulminant MH crisis induced by isoflurane anesthesia and rescues animals from heat stroke caused by environmental heat stress. Notably, Cpd1 has great advantages of better water solubility and rapid clearance in vivo over dantrolene. Cpd1 has the potential to be a promising candidate for effective treatment of patients carrying RyR1 mutations., Mutations in ryanodine receptor 1 (RyR1), a Ca2+ release channel in skeletal muscle, cause malignant hyperthermia (MH) and are involved in heat stroke. Here, the authors show that an oxolinic acid-derivative RyR1 inhibitor effectively prevents and treats MH and heat stroke in various MH mouse models.

The chylous turbidity of blood samples is one of the causes of false-high hemoglobin (Hgb) concentration measurements by the colorimetric method, which has been widely applied in hematology analyzers. In such cases, additional manual procedures are required to correct Hgb concentrations. We therefore examined the effectiveness of an optical method for measuring Hgb concentrations in samples with chylous turbidity using Hgb-O in the reticulocyte channel equipped in XN-series analyzers (Sysmex, Kobe, Japan). Hgb-O showed excellent basic performance, including linear correlation and invariability with sodium lauryl sulfate (SLS)-Hgb detected by the colorimetric method. In the analysis of samples from healthy volunteers supplemented with fat emulsion, chylous turbidity did not affect Hgb-O but SLS-Hgb, which was falsely increased according to the dose of fat emulsion. Actually, SLS-Hgb was falsely elevated in 34 of 40 chylous turbidity 3+ samples. The remaining 6 samples were measured in hematology analyzers where Hgb-O was inconsistent with SLS-Hgb in the internal quality control records. For these samples, the correction factors calculated from the internal quality control records could contribute to providing the corrected Hgb-O value. These findings suggested that the optical method was effective and convenient for accurately evaluating Hgb concentrations in samples with extremely chylous turbidity.

BACKGROUND Children with Down syndrome (DS) have different growth rates compared with normal children. The present study examined the reliability of a general formula, Uemura's formula, utilized in normal Japanese children to estimate renal function (estimated glomerular filtration rate - eGFR) in children with DS. METHODS This study included 758 children aged 2-18 years with DS who visited our medical center. Patients with congenital heart disease, or congenital anomalies of the kidney or urinary tract detected via abdominal ultrasonography, chronic glomerulonephritis, and vesicoureteral reflux, etc., were excluded. Height and serum creatinine data gathered from 2421 examinations of 379 children with DS (224 boys and 155 girls) were used to evaluate Uemura's formula. RESULTS The mean eGFR was lower in children with DS than in children without DS. Stage II chronic kidney disease was indicated in 44.6% of examinations and stage III in 0.8%. The association of eGFR with age differed between sexes. Boys with DS showed a significant but weak negative correlation between eGFR and age (r = -0.273, P

Positron emission tomography (PET) has been used to investigate changes in the concentration of endogenous neurotransmitters. Recently, this technique has been applied to the imaging of serotonin2A receptors using [18F]altanserin. In these measurements, a reduction in binding potential (BP) suggests an increase in endogenous serotonin levels caused by pharmacological or cognitive stimulations, and the sensitivity of BP reduction depends on the characteristics of [18F]altanserin. In this study, we evaluated an analytical method for estimating the changes in endogenous serotonin levels based on PET scans with [18F]altanserin at baseline and stimulated states and validated it using simulations and small animal PET studies. First, in the simulations, the time-activity curves at baseline and the stimulated states were generated using an extended compartment model including the competition for the receptors between the administered [18F]altanserin and endogenous serotonin. In the stimulated state, the magnitude and onset of the endogenous serotonin elevation were altered to varying degrees. In these time-activity curves, BP was estimated using the simplified reference tissue model (SRTM), and the reduction in BP was evaluated by comparison with that of the baseline state. Next, the proposed method was applied to mouse PET studies. Endogenous serotonin levels were elevated by treatment with selective serotonin reuptake inhibitors (SSRIs), and PET studies were performed twice, once with and once without treatment. In both scans, BP was estimated using the SRTM with the cerebellum as a reference region, and the reduction in BP after SSRI treatment was evaluated. In the simulations, the BP estimate of the stimulated state was smaller than that of the baseline state, and their reduction was related to the amount of change in the serotonin concentration. BP reduction was also affected by the onset of serotonin elevation. In the mouse studies, the BP of the cerebral cortex decreased in the scans with SSRI treatment. The reduction in BP estimated using the SRTM from [18F]altanserin-PET studies at baseline and in stimulated states can detect changes in the binding conditions of serotonin2A receptors. This may be useful for investigating the elevation of endogenous serotonin levels caused by stimulations.

Extracellular vesicles (EVs) are secreted from most cells and play important roles in cell-cell communication by transporting proteins, lipids, and nucleic acids. As the involvement of EVs in diseases has become apparent, druggable regulators of EV secretion are more desirable. However, the lack of a highly sensitive EV detection system has made the development of EV regulators difficult. We developed an ELISA system to detect EVs using TIM4 proteins, which have high affinity to phosphatidylserine and screened a 1,567-compound library. Consequently, we identified one inhibitor and three activators of EV secretion in a variety of cells. The inhibitor, apoptosis activator 2, suppressed EV secretion via a different mechanism and had a broader cellular specificity than GW4869. The three activators, cucurbitacin B, gossypol, and obatoclax, also had broad cellular specificity, including HEK293T cells and human mesenchymal stem cells (hMSCs). In vitro bioactivity assays revealed that some regulators control EV secretion from glioblastoma and hMSCs, which induces angiogenesis and protects cardiomyocytes against apoptosis, respectively. In conclusion, we developed a high-throughput method to detect EVs with high sensitivity and versatility, and identified three compounds that can regulate the bioactivity of EVs.

Background Prognosticating disease progression in patients with diabetic kidney disease (DKD) is challenging, especially in the early stages of kidney disease. Anemia can occur in the early stages of kidney disease in diabetes. We therefore postulated that serum hemoglobin (Hb) concentration, as a reflection of incipient renal tubulointerstitial impairment, can be used as a marker to predict DKD progression. Methods Drawing on nationally representative data of patients with biopsy-proven DKD, 246 patients who had an estimated glomerular filtration rate (eGFR) ≥60 mL/min/1.73 m2 at renal biopsy were identified: age 56 (45–63) years; 62.6% men; Hb 13.3 (12.0–14.5) g/dL; eGFR 76.2 (66.6–88.6) mL/min/1.73 m2; urine albumin-to-creatinine ratio 534 (100–1480) mg/g Crea. Serum Hb concentration was divided into quartiles: ≤12, 12.1–13.3, 13.4–14.5 and ≥14.6 g/dL. The association between serum Hb concentration and the severity of renal pathological lesions was explored. A multivariable Cox regression model was used to estimate the risk of DKD progression (new onset of end-stage kidney disease, 50% reduction of eGFR or doubling of serum creatinine). The incremental prognostic value of DKD progression by adding serum Hb concentration to the known risk factors of DKD was assessed. Results Serum Hb levels negatively correlated with all renal pathological features, especially with the severity of interstitial fibrosis (ρ = −0.52; P Conclusions Serum Hb concentration, which reflects incipient renal fibrosis, can be useful for predicting DKD progression in the early stages of kidney disease.

In this work, the design, synthesis, and single-molecule conductance of ethynyl- and butadiynyl-ruthenium molecular wires with thioether anchor groups [RS=n-C6 H13 S, p-tert-Bu-C6 H4 S), trans-{RS-(C≡C)n }2 Ru(dppe)2 (n=1 (1R ), 2 (2R ); dppe: 1,2-bis(diphenylphosphino)ethane) and trans-(n-C6 H13 S-C≡C)2 Ru{P(OMe)3 }4 3hex ] are reported. Scanning tunneling microscope break-junction study has revealed conductance of the organometallic molecular wires with the thioacetylene backbones higher than that of the related organometallic wires having arylethynylruthenium linkages with the sulfur anchor groups, trans-{p-MeS-C6 H4 -(C≡C)n }2 Ru(phosphine)4 4n (n=1, 2) and trans-(Th-C≡C)2 Ru(phosphine)4 5 (Th=3-thienyl). It should be noted that the molecular junctions constructed from the butadiynyl wire 2R , trans-{Au-RS-(C≡C)2 }2 Ru(dppe)2 (Au: gold metal electrode), show conductance comparable to that of the covalently linked polyynyl wire with the similar molecular length, trans-{Au-(C≡C)3 }2 Ru(dppe)2 63 . The DFT non-equilibrium Green's function (NEGF) study supports the highly conducting nature of the thioacetylene molecular wires through HOMO orbitals.

In mammalian cloning by somatic cell nuclear transfer (SCNT), the treatment of reconstructed embryos with histone deacetylase (HDAC) inhibitors improves efficiency. So far, most of those used for SCNT are hydroxamic acid derivatives—such as trichostatin A—characterized by their broad inhibitory spectrum. Here, we examined whether mouse SCNT efficiency could be improved using chlamydocin analogues, a family of newly designed agents that specifically inhibit class I and IIa HDACs. Development of SCNT-derived embryos in vitro and in vivo revealed that four out of five chlamydocin analogues tested could promote the development of cloned embryos. The highest pup rates (7.1–7.2%) were obtained with Ky-9, similar to those achieved with trichostatin A (7.2–7.3%). Thus, inhibition of class I and/or IIa HDACs in SCNT-derived embryos is enough for significant improvements in full-term development. In mouse SCNT, the exposure of reconstructed oocytes to HDAC inhibitors is limited to 8–10 h because longer inhibition with class I inhibitors causes a two-cell developmental block. Therefore, we used Ky-29, with higher selectivity for class IIa than class I HDACs for longer treatment of SCNT-derived embryos. As expected, 24-h treatment with Ky-29 up to the two-cell stage did not induce a developmental block, but the pup rate was not improved. This suggests that the one-cell stage is a critical period for improving SCNT cloning using HDAC inhibitors. Thus, chlamydocin analogues appear promising for understanding and improving the epigenetic status of mammalian SCNT-derived embryos through their specific inhibitory effects on HDACs., Chlamydocin analogues, a novel family of inhibitors specific for class I and IIb HDACs, significantly improved the ability of mouse SCNT-derived embryos to produce offspring.

Aims The impacts of high density lipoprotein cholesterol (HDL‐C) as an anti‐inflammatory and C reactive protein (CRP) as inflammatory properties on the pathogenesis of heart failure were reported. At present, the clinical significance of the HDL‐C/CRP ratio in heart failure with preserved ejection fraction (HFpEF) patients remains unknown. Methods and results We examined the data on 796 consecutive HFpEF (left ventricular ejection fraction ≥50%) patients hospitalized due to acute decompensated heart failure from the PURSUIT‐HFpEF registry, a prospective, multicentre observational study. We calculated the HDL/CRP ratios and evaluated the relationship between the values and clinical outcomes, including degree of cardiac function. The mean follow‐up duration was 420 ± 346 days. All‐cause death occurred in 118 patients, of which 51 were cardiac deaths. HDL/CRP ≤ 4.05 was independently and significantly associated with all‐cause death (odds ratio = 1.84, 95% CI: 1.06–3.20, P = 0.023), and HDL/CRP ≤ 3.14 was associated with cardiac death by multivariate Cox proportional hazard analysis (odds ratio = 2.86, 95% CI: 1.36–6.01, P = 0.003). HDL‐C/CRP ratio significantly correlated with the product of the left atrial volume and left ventricular mass index as well as the tricuspid annular plane systolic excursion by multiple regression analysis (standardized beta‐coefficient = −0.085, P = 0.034 and standardized beta‐coefficient = 0.081, P = 0.044, respectively). Conclusions HDL‐C/CRP ratio was a useful marker for predicting all‐cause death and cardiac death and correlated with left ventricular diastolic function and right ventricular systolic function in HFpEF patients.

The 70% ethanol extract of Artemisia campestris was screened to find PPARγ ligands using the PPARγ ligand-responsive chimera luciferase reporter system. Capillartemisin B was identified as a PPARγ ligand that stimulated lipid accumulation in 3T3-L1 cells. By further purification of PPARγ ligands from a large-scale preparation of the methanol extract of Artemisia campestris, we isolated and identified eupatilin and santaflavone as PPARγ ligands. Weak PPARγ ligand activity of eupatilin or santaflavone in reporter assay was enhanced by a PPARγ antagonist, GW9662, suggesting that santaflavone or eupatilin and GW9662 bound simultaneously to the multiple sub-pockets of the PPARγ ligand-binding domain (LBD) and cooperatively activated PPARγ. Docking simulation suggested that eupatilin binds to the Ω-pocket but not to the AF-2 pocket of Y-shaped PPARγ LBD where artepillin C that differs from capillartemisin B at the C-5′ position without hydroxy group binds. Eupatilin or santaflavone with or without GW9662 did not stimulate lipid accumulation in differentiated 3T3-L1 cells, suggesting that binding of each compound alone or with GW9662 to the Ω-pocket which stimulated the PPARγ-responsive reporter expression was not enough to stimulate lipid accumulation. The PPARγ ligands found in this study have a potential to design the fragment-based drug design of a novel PPARγ ligand that cover the Y-shaped PPARγ LBD.

We evaluated the in vitro drug–drug interaction (DDI) potential of enerisant (TS-091), a histamine H3 receptor antagonist/inverse agonist, mediated by cytochrome P450 (CYP) and transporters, as wel...

Glandular epithelial cells (GE) in the endometrium are thought to support the elongation and survival of ruminant embryos by secreting histotrophs. In the present study, the gene expression of bovine endometrial epithelial cells cultured in matrigel was analyzed and examined whether it could be an in vitro model of GE. Bovine endometrial epithelial cells (BEE) and stromal cells (BES) were isolated from the slaughterhouse uteri and cultured in DMEM/F12 + 10% FBS. BEE showed the gland-like structure morphological changes when cultured in 15% matrigel but could not be identified in higher concentrations of the matrigel (30% or 60%). The expression of typical genes expressed in GE, SERPINA14 and GRP, was substantially high in matrigel-cultured BEE than in monolayer (P

We investigated a single-molecule reaction of DNA intercalation as an example of a bimolecular association reaction. Single-molecule conductance values of the product and reactant molecules adsorbed on an Au surface were measured to identify and quantify these molecules. The binding isotherm was constructed, and the association constant of the reaction was determined on a single-molecule basis.

Background In mammals, there are two subtypes of Grb2-associated regulator of Erk/MAPK (GAREM), an adaptor protein that functions downstream of the cell growth factor receptor. GAREM1 is ubiquitously expressed, whereas GAREM2 is mainly expressed in the brain. However, the precise mechanism of the translocation of each GAREM subtype in growth factor-stimulated cells is still unclear. Methods In this study, immunofluorescence staining with specific antibodies against each GAREM subtype and time-lapse analysis using GFP fusion proteins were used to analyze the subcellular localization of each GAREM subtype in a cell growth stimulus-dependent manner. We also biochemically analyzed the correlation between its subcellular localization and tyrosine phosphorylation of GAREM2. Results We found that endogenously and exogenously expressed GAREM2 specifically aggregated and formed granules in NGF-stimulated PC-12 cells and in EGF-stimulated COS-7 cells. Based on the observed subcellular localizations of chimeric GAREM1 and GAREM2 proteins, a glycine-rich region, which is present only in GAREM2, is required for the observed granule formation. This region also regulates the degree of EGF-stimulation-dependent tyrosine phosphorylation of GAREM2. Conclusions Our results, showing that aggregation of GAREM2 in response to EGF stimulation is dependent on a glycine-rich region, suggest that GAREM2 aggregation may be involved in neurodegenerative diseases.

Multidrug efflux pumps are inner membrane transporters that export multiple antibiotics from the inside to the outside of bacterial cells, contributing to bacterial multidrug resistance (MDR). Postgenomic analysis has demonstrated that numerous multidrug efflux pumps exist in bacteria. Also, the co-crystal structural analysis of multidrug efflux pumps revealed the drug recognition and export mechanisms, and the inhibitory mechanisms of the pumps. A single multidrug efflux pump can export multiple antibiotics; hence, developing efflux pump inhibitors is crucial in overcoming infectious diseases caused by multidrug-resistant bacteria. This review article describes the role of multidrug efflux pumps in MDR, and their physiological functions and inhibitory mechanisms.

Dialkyl-diphosphenes and -distibenes of the type RTrp*2E2 (E = P, Sb; R = H, n-Pr) were synthesized and isolated using the steric protection of extended triptycyl groups (Trp*). The solubility of t...

Advanced ovarian clear cell carcinoma (OCCC) is a recalcitrant disease, often resistant to the first-line platinum-based therapy. Using a novel patient-derived orthotopic xenograft (PDOX) nude-mouse model of OCCC, we tested whether oral-recombinant methioninase (o-rMETase) could enhance the efficacy of paclitaxel (PTX). The OCCC PDOX model was established and passaged in nude mice. The OCCC PDOX models were randomized into 5 groups. G1: untreated control; G2: paclitaxel (PTX) (20 mg/kg, intraperitoneal (i.p.) injection, weekly); G3: o-rMETase (100 units, oral, daily); G4: PTX (20 mg/kg, i.p. injection, weekly) + carboplatinum (CBDCA) (40 mg/kg, i.p. injection weekly); G5: PTX (20 mg/kg, i.p. injection, weekly) + o-rMETase (100 units, oral, daily). The treatment period was 2 weeks. The combination of PTX and o-rMETase arrested OCCC tumor growth (relative tumor volume: 1.09 ± 0.63 (mean ± SD)) compared with the untreated control (relative tumor volume: 3.92 ± 1.04 (mean ± SD)) (p

Heat shock protein family B member 8, encoded by HSPB8, is an essential component of the chaperone-assisted selective autophagy complex, which maintains muscle function by degrading damaged proteins in the cells. Mutations in HSPB8 have been reported to cause Charcot-Marie-Tooth type 2L, distal hereditary motor neuropathy IIa, and rimmed vacuolar myopathies (RVM). In this study, we identified a novel heterozygous frameshift variant c.525_529del in HSPB8 in a large Japanese family with RVM, using whole exome sequencing. Three affected individuals had severe respiratory failure, which has not been addressed by previous studies. Muscle atrophy in the paraspinal muscles was also a clinical feature of the individuals affected with RVM in this study. The frameshift mutation was located in the last coding exon, and the mutated protein was predicted to harbor an isoleucine-leucine-valine (ILV) sequence, which corresponds to the IXI/V (isoleucine, X amino acids, and isoleucine or valine) motif. The IXI/V motif is essential for assembly into larger oligomers in other small heat shock proteins and all frameshift mutants of HSPB8 were predicted to share the ILV sequence in the C-terminal extension. The in silico prediction tools showed low protein solubility and increased aggregation propensity for the region around the ILV sequence. The IXI/V motif might be associated with the pathogenesis of HSPB8-related RVM.

BackgroundThe standard treatment for stage IB–IIB cervical cancer is radiotherapy or radical hysterectomy; after radical hysterectomy, adjuvant concurrent chemoradiotherapy is recommended for patients with high risk factors. However, adjuvant concurrent chemoradiotherapy can cause severe gastrointestinal and urinary toxicity.Primary ObjectiveTo assess whether postoperative adjuvant chemotherapy is not inferior to adjuvant concurrent chemoradiotherapy for overall survival in patients with high risk cervical cancer.Study HypothesisAdjuvant chemotherapy is not inferior to adjuvant concurrent chemoradiotherapy for overall survival and will reduce severe toxicities.Trial DesignPatients with high risk factors after radical hysterectomy will be randomized 1:1 to receive adjuvant concurrent chemoradiotherapy or adjuvant chemotherapy. Treatment will be started within 6 weeks of surgery. The concurrent chemoradiotherapy group will receive whole pelvis irradiation (50.4 Gy) and cisplatin (40 mg/m2/week). The chemotherapy group will receive paclitaxel (175 mg/m2) plus cisplatin (50 mg/m2) or carboplatin (AUC=6) every 3 weeks for six cycles.Major Inclusion/Exclusion CriteriaPatients with high risk stage IB–IIB cervical cancer (squamous cell carcinoma, adenocarcinoma, and adenosquamous cell carcinoma) who underwent radical hysterectomy are eligible for the study. High risk is defined as the presence of pelvic lymph node metastasis and/or parametrial invasion.Primary EndpointThe primary endpoint is overall survival.Sample Size250 patients in total are required.Estimated Dates for Completing AccrualThis study began in November 2019, and 250 patients will be accrued within 5 years.Trial Registration NumberThe study has been registered with the Japan Registry of Clinical Trials (jRCTs041190042).

Purpose: In ALK-rearranged non–small cell lung cancer (NSCLC), impacts of concomitant genetic alterations on targeted therapies with ALK-tyrosine kinase inhibitors (ALK-TKI) are not yet well understood. Here, we investigated genetic alterations related to ALK-TKI resistance using clinico-genomic data and explored effective therapies to overcome the resistance in preclinical models through the identification of underlying molecular mechanisms. Experimental Design: We used integrated clinical and next-generation sequencing data generated in a nationwide lung cancer genome screening project (LC-SCRUM-Japan). ALK-rearranged NSCLC cell lines expressing wild-type or mutant TP53 were used to evaluate cellular apoptosis induced by ALK-TKIs. Results: In 90 patients with ALK-rearranged NSCLC who were treated with a selective ALK-TKI, alectinib, TP53 comutated patients showed significantly worse progression-free survival (PFS) than TP53 wild-type patients [median PFS, 11.7 months (95% confidence interval, CI, 6.3–not reached, NR) vs. NR (23.6–NR); P = 0.0008; HR, 0.33 (95% CI, 0.17–0.65)]. ALK-rearranged NSCLC cell lines that lost p53 function were resistant to alectinib-induced apoptosis, but a proteasome inhibitior, ixazomib, markedly induced apoptosis in the alectinib-treated cells by increasing the expression of a proapoptotic protein, Noxa, which bound to an antiapoptotic protein, Mcl-1. In subcutaneous tumor models, combination of ixazomib and alectinib prominently induced tumor regression and apoptosis even though the tumors were generated from ALK-rearranged NSCLC cells with nonfunctional p53. Conclusions: These clinical and preclinical results indicate concomitant TP53 mutations reduce the efficacy of alectinib for ALK-rearranged NSCLC and the combined use of a proteasome inhibitor with alectinib is a promising therapy for ALK-rearranged/TP53-mutated NSCLC.

Hitachimycin is a macrolactam antibiotic with an (S)-β-phenylalanine (β-Phe) at the starter position of its polyketide skeleton. (S)-β-Phe is formed from l-α-phenylalanine by the phenylananine-2,3-aminomutase HitA in the hitachimycin biosynthetic pathway. In this study, we produced new hitachimycin analogs via mutasynthesis by feeding various (S)-β-Phe analogs to a ΔhitA strain. We obtained six hitachimycin analogs with F at the ortho, meta, or para position and Cl, Br, or a CH3 group at the meta position of the phenyl moiety, as well as two hitachimycin analogs with thienyl substitutions. Furthermore, we carried out a biochemical and structural analysis of HitB, a β-amino acid-selective adenylation enzyme that introduces (S)-β-Phe into the hitachimycin biosynthetic pathway. The KM values of the incorporated (S)-β-Phe analogs and natural (S)-β-Phe were similar. However, the KM values of unincorporated (S)-β-Phe analogs with Br and a CH3 group at the ortho or para position of the phenyl moiety were high, indicating that HitB functions as a gatekeeper to select macrolactam starter units during mutasynthesis. The crystal structure of HitB in complex with (S)-β-3-Br-phenylalanine sulfamoyladenosine (β-m-Br-Phe-SA) revealed that the bulky meta-Br group is accommodated by the conformational flexibility around Phe328, whose side chain is close to the meta position. The aromatic group of β-m-Br-Phe-SA is surrounded by hydrophobic and aromatic residues, which appears to confer the conformational flexibility that enables HitB to accommodate the meta-substituted (S)-β-Phe. The new hitachimycin analogs exhibited different levels of biological activity in HeLa cells and multidrug-sensitive budding yeast, suggesting that they may target different molecules.

In this work, the δ%Y–10%Co/MA (δ = 0, 1, 2, 3, and 5 wt%) catalysts synthesised by sequential incipient wetness impregnation were characterised and evaluated in CH4 dry reforming. Superior catalytic performances were shown by 3 wt% Y2O3 loading (CH4 conversion = 85.8%, and CO2 conversion = 90.5%), followed by 2 wt% > 5 wt% > 1 wt% > 0 wt% Y2O3 loading. This result was attributed to the favorable catalytic properties of 3 wt%Y–10%Co/MA including small Co particle size, high Co dispersion, high amount of atomic ratio (Co/Al), and high number of lattice oxygen vacancies. The excess Y2O3 addition (>3 wt%) led to inevitably blocked Co active sites and resulted in decreasing catalytic performance. The 3 wt% Y2O3 promoter loading recorded the lowest carbon deposited (7.0%) due to the highest oxygen vacancies (78.1%) compared to 1, 2 and 5 wt% Y2O3.

In this study, general methods were applied for the preparation of new iminopyridazine butyronitriles. A series of six new 4-(3-aryl-1,6-dihydro-6-iminopyridazin-1-yl)butyronitrile hydro-bromides (2a-2f) (Scheme 1) have been prepared starting from commercially available 3-amino-6-chloropyridazine in two steps with good yields. The synthesized compound’s structures were characterized by IR, 1H NMR and high-resolution mass structures spectral (HRMS) data. Journal of Bangladesh Academy of Sciences, Vol. 44, No. 2, 131-138, 2020

We investigated the electronic structure and charge transport properties of single-molecule junctions of a series of acene-type molecules (i.e., pyrazine, quinoxaline, and phenazine) with different...

A 69-year-old woman was admitted to our hospital because of limb weakness. She was diagnosed to have chronic renal failure due to diabetes mellitus and had suffered from pericardial effusion at 67 years of age. She started taking colchicine 18 months before admission and thereafter gradually developed muscle weakness in her limbs and had become bedridden at the time of admission. The withdrawal of colchicine improved her limb weakness, and therefore we diagnosed her to have colchicine myopathy. Her muscle strength did not completely recover even after six months from cessation of colchicine. It was suggested that renal failure and muscle disuse had prevented the full recovery of her muscles in addition to the long-term use of colchicine. Typical colchicine myopathy improves rapidly, but the long-term use of colchicine is considered to cause muscle weakness. Although the CK level was elevated, the elevated CK and myopathy had been overlooked because the CK baseline was low due to the patient's small amount of muscle mass. Moreover, the estimated GFR was recorded to be higher than her actual renal function due to her small amount of muscle mass, therefore the risk of colchicine myopathy in this case remained unrecognized.

Three crystals of MHF and its complex with FANCM were obtained, and structural analysis revealed not only their structures but the unexpected release of FANCM from MHF. The biochemical stability of the FANCM–MHF complex was analyzed and it was found that an oxidative environment and organic solvent promote the aggregation and dissociation of the complex., FANCM is involved in eukaryotic DNA-damage recognition and activates the Fanconi anemia (FA) pathway through complex formation. MHF is one of the FANCM-associating components and contains a histone-fold DNA-binding domain. Loss of the FANCM–MHF interaction compromises the activation of the FA pathway, resulting in chromosomal instability. Thus, formation of the FANCM–MHF complex is important for function, but its nature largely remains elusive. Here, the aim was to reveal the molecular and structural basis for the stability of the FANCM–MHF complex. A recombinant tripartite complex containing chicken FANCM (MHF interaction region), MHF1 and MHF2 was expressed and purified. The purified tripartite complex was crystallized under various conditions and three different crystals were obtained from similar crystallization conditions. Unexpectedly, structure determination revealed that one of the crystals contained the FANCM–MHF complex but that the other two contained the MHF complex without FANCM. How FANCM dissociates from MHF was further investigated and it was found that the presence of 2-methyl-2,4-pentanediol (MPD) and an oxidative environment may have promoted its release. However, under these conditions MHF retained its complexed form. FANCM–MHF interaction involves a mixture of hydrophobic/hydrophilic interactions, and chicken FANCM contains several nonconserved cysteines within this region which may lead to aggregation with other FANCM–MHF molecules. These results indicate an unexpected nature of the FANCM–MHF complex and the data can be used to improve the stability of the complex for biochemical and structural analyses.

Background Diabetes mellitus is the most commonly encountered endocrinopathy in patients with mitochondrial encephalopathy, lactic acidosis, and stroke-like episodes (MELAS), which manifests as multisystemic organ failure. Whether the management of diabetes mellitus in MELAS requires special consideration is not fully clarified. Methods In this single-center study, we retrospectively reviewed the medical records of patients with MELAS to elucidate the clinical characteristics of MELAS–associated diabetes mellitus. Results Four patients among a total of 25 individuals with MELAS who were treated in the study institution developed diabetes mellitus. One patient had well-controlled diabetes mellitus, whereas the remaining three patients experienced hyperglycemic crisis as the first manifestation of diabetes mellitus. Two of the three patients were children aged four and six years. The hyperglycemic events occurred after surgery, infection, and status epilepticus, respectively. None of the three patients had diabetes mellitus previously based on randomly measured serum glucose levels that were within the normal range before the hyperglycemic crisis. Glycated hemoglobin levels measured during the hyperglycemic crisis indicated prediabetes in two patients and diabetes mellitus in one patient. Two patients recovered, whereas one patient died after developing multiorgan failure. Conclusions Fulminant-onset diabetes mellitus occurring in patients with MELAS underscore the importance of routine measurement for glycated hemoglobin and more intense evaluation of glucose intolerance regardless of the patient age and lack of symptoms. Clinicians should be aware of the potential acute onset of hyperglycemic crisis in patients with MELAS, especially in individuals with aggravating factors.

A weak boundary layer (WBL), at the surface of polymers, causes poor adhesion, but such a WBL has not been characterized or quantified. In this study, to characterize a WBL of polytetrafluoroethylene (PTFE), we observed a cross-section of PTFE using a scanning electron microscope and compared the surfaces and bulk sides. We observed voids on the surface of the PTFE but not on the bulk side. These voids existed up to 5 µm from the outermost surface of the PTFE. We defined a WBL by the presence or absence of voids and then concluded that the thickness of the WBL in PTFE was of the order of a single micrometer. Polytetrafluoroethylene (PTFE) has a weak boundary layer (WBL) on the surface. A cross-section of PTFE was observed using a scanning electron microscope for investigating the difference in morphology between WBL and bulk layer. Large voids of 0.5–2.0 µm in diameter were observed on the surface side of PTFE but not on the bulk side. These voids existed up to approximately 5 µm from the outermost surface of the PTFE. This result indicated that the thickness of WBL of PTFE was in the order of single µm.

Poly(vinyl butyral) (PVB) is widely accepted as an adhesive for glass substrates within the automobile windows. In this work, we suggested and performed butyralization of poly(vinyl alcohol) (PVA) under supercritical carbon dioxide (sc-CO2) and investigated not only the structure and mechanical properties of the obtained PVB but also its adhesion properties under various conditions, comparing this PVA with other PVBs prepared in the solution and swollen states. The conversion ratio of butyralization under sc-CO2 was larger than that in the swollen state and lower than that in the solution state and exhibited a sufficient material performance as the adhesive for glass substrates. The Young’s modulus and tensile strength of PVB under sc-CO2 were higher than those of the other PVBs. The mechanical properties of the PVB prepared under sc-CO2 have no correlation to the modification ratios because sc-CO2 penetrated into the amorphous region of the PVA and preferentially modified its hydroxyl groups. Furthermore, the adhesive strengths of all the obtained PVBs increased, and under a high-humidity atmosphere, the adhesive strength of the PVB prepared under sc-CO2 was the largest. The humidity resistance of the PVB adhesive prepared under sc-CO2 was proven. Poly(vinyl butyral) (PVB) is accepted as an adhesive to glass substrates. The PVB was prepared through butyralization of poly(vinyl alcohol) under supercritical carbon dioxides. The Young’s modulus and tensile strength of the PVB prepared under supercritical carbon dioxides were higher than those of the PVBs in the preparation method in solution and swollen states. In addition, the PVB under supercritical carbon dioxide possessed higher adhesive strength to glass substrates even under high humid condition, compared with the other PVBs.

Background/purpose Indocyanine green (ICG) fluorescence navigation has been adapted for anatomic liver resection (AR) but an objective method for evaluation of its validity is required. This pilot study aimed to propose a new method to evaluate the accuracy of parenchymal division along the plane between hepatic segments and estimate the real-time navigation efficacy for AR by the Medical Imaging Projection System (MIPS), which continuously demonstrates the transection plane using projection mapping with ICG fluorescence. Methods Ten patients who underwent open AR using liver segmentation with ICG fluorescence technique between August 2016 and July 2019 were included: six patients under MIPS guidance (MIPS group), while four using only conventional ICG fluorescence technique before parenchymal resection (non-MIPS group). Densitometry of the captured fluorescence image was performed to evaluate the fluorescence area ratio of each transection plane. The accurate fluorescence area ratio was calculated by subtracting the fluorescence area rate on the resected side from that on the remnant side. Results The accurate fluorescence area ratio of the MIPS group and the non-MIPS group was 23.0 ± 12.6% and 5.6 ± 9.5%, respectively (P = .038). Conclusions Based on the results of our new method, real-time navigation using the MIPS may facilitate performing AR along the plane between hepatic segments.

We describe the development of a one-pot preparation process of (E)-3,5-dichloro-4-(3-ethoxy-2-methyl-3-oxoprop-1-en-1-yl)benzoic acid (3), which is a key carboxylic acid intermediate of lusutrombo...

Vasohibin-1 and small vasohibin-binding protein (SVBP) form an intermolecular heterotetramer in the crystal. The heterotetramer was stabilized by exchange of the conserved N-terminal region., Vasohibins regulate angiogenesis, tumor growth, metastasis and neuronal differentiation. They form a complex with small vasohibin-binding protein (SVBP) and show tubulin tyrosine carboxypeptidase activity. Recent crystal structure determinations of vasohibin–SVBP complexes have provided a molecular basis for complex formation, substrate binding and catalytic activity. However, the regulatory mechanism and dynamics of the complex remain elusive. Here, the crystal structure of the VASH1–SVBP complex and a molecular-dynamics simulation study are reported. The overall structure of the complex was similar to previously reported structures. Importantly, however, the structure revealed a domain-swapped heterotetramer that was formed between twofold symmetry-related molecules. This heterotetramerization was stabilized by the mutual exchange of ten conserved N-terminal residues from the VASH1 structural core, which was intramolecular in other structures. Interestingly, a comparison of this region with previously reported structures revealed that the patterns of hydrogen bonding and hydrophobic interactions vary. In the molecular-dynamics simulations, differences were found between the heterotetramer and heterodimer, where the fluctuation of the N-terminal region in the heterotetramer was suppressed. Thus, heterotetramer formation and flexibility of the N-terminal region may be important for enzyme activity and regulation.

Recently, smaller-size electron-beam (EB) accelerators have offered EB irradiation in laboratory systems. Therefore, polymer surface treatments with low-energy EB have been developed in the past years. For high adhesion strength, low-energy EB treatment is also a promising method in comparison to plasma surface treatment. In the plasma treatment, the mechanism of the effect on the adhesion properties has been proved and the excess treatments led to the formation of a weak boundary layer and reduction of adhesion strength. In contrast, the low-energy EB possesses high penetration ability. In this work, we focused on the surface treatments of isotactic polypropylene (it.PP) with low-energy EB irradiation for adhesion. The dependence of adhesion strength on the absorbed dose of electron beam was evaluated, and the mechanism of electron beam on the adhesion properties was investigated from various perspectives of surface properties and morphology. Compared to that of plasma-treated it.PP, the adhesion strength of it.PP with electron-beam irradiation increased drastically. We proved that the radical was generated in the substrates after electron-beam treatments and would form covalent bonds between adhesives and substrates, which achieved higher adhesion than plasma treatments. In addition, the electron beam reached effectively a deep region from the top surface of the substrates and provided larger adhesion strength.