Isolation and characterization ofβ-transducin repeat-containing protein ligands screened using a high-throughput screening system

β-transducin repeat-containing protein(β-TrCP)is an F-box protein subunit of the E3 Skp1-Cullin-F box(SCF)type ubiquitin-ligase complex,and provides the substrate specificity for the ligase.To find potent ligands ofβ-TrCP useful for the proteolysis targeting chimera(PROTAC)system usingβ-TrCP in the future,we developed a high-throughput screening system for small moleculeβ-TrCP ligands.We screened the chemical library utilizing the system and obtained several hit compounds.The effects of the hit compounds on in vitro ubiquitination activity of SCFβ-TrCP1 and on downstream signaling pathways were examined.Hit compounds NPD5943,NPL62020-01,and NPL42040-01 inhibited the TNFα-induced degradation of IκBαand its phosphorylated form.Hence,they inhibited the activation of the transcription activity of NF-κB,indicating the effective inhibition ofβ-TrCP by the hit compounds in cells.Next,we performed an in silico analysis of the hit compounds to determine the important moieties of the hit compounds.Carboxyl groups of NPL62020-01 and NPL42040-01 and hydroxyl groups of NPD5943 created hydrogen bonds withβ-TrCP similar to those created by intrinsic target phosphopeptides ofβ-TrCP.Our findings enhance our knowledge of useful small molecule ligands ofβ-TrCP and the importance of residues that can be ligands ofβ-TrCP.

Identification of a dihydroorotate dehydrogenase inhibitor thatinhibits cancer cell growth by proteomic profiling

Dihydroorotate dehydrogenase(DHODH)is a central enzyme of the de novo pyrimidine biosynthesis pathway and is a promising drug target for the treatment of cancer and autoimmune diseases.This study presents the identification of a potent DHODH inhibitor by proteomic profiling.Cell-based screening revealed that NPD723,which is reduced to H-006 in cells,strongly induces myeloid differentiation and inhibits cell growth in HL-60 cells.H-006 also suppressed the growth of various cancer cells.Proteomic profiling of NPD723-treated cells in ChemProteoBase showed that NPD723 was clustered with DHODH inhibitors.H-006 potently inhibited human DHODH activity in vitro,whereas NPD723 was approximately 400 times less active than H-006.H-006-induced cell death was rescued by the addition of the DHODH product orotic acid.Moreover,metabolome analysis revealed that H-006 treatment promotes marked accumulation of the DHODH substrate dihydroorotic acid.These results suggest that NPD723 is reduced in cells to its active metabolite H-006,which then targets DHODH and suppresses cancer cell growth.Thus,H-006-related drugs represent a potentially powerful treatment for cancer and other diseases.

The 2023 Turkey earthquake doublet: Earthquake relocation, seismic tomography, and stress field inversion

On February 6,2023,two earthquakes with magnitudes of M_(W) 7.8 and M_(W) 7.5 struck southeastern Turkey,causing significant casualties and economic losses.These seismic events occurred along the East Anatolian Fault Zone,a convergent boundary between the Arabian Plate and the Anatolian Subplate.In this study,we analyze the M_(W) 7.8 and M_(W) 7.5 earthquakes by comparing their aftershock relocations,tomographic images,and stress field inversions.The earthquakes were localized in the upper crust and exhibited steep dip angles.Furthermore,the aftershocks occurred either close to the boundaries of low and high P-wave velocity anomaly zones or within the low P-wave velocity anomaly zones.The East Anatolia Fault,associated with the M_(W) 7.8 earthquake,and the SürgüFault,related to the M_(W) 7.5 earthquake,predominantly experienced shear stress.However,their western sections experienced a combination of strike-slip and tensile stresses in addition to shear stress.The ruptures of the M_(W) 7.8 and M_(W) 7.5 earthquakes appear to have bridged a seismic gap that had seen sparse seismicity over the past 200 years prior to the 2023 Turkey earthquake sequence.

Inhibitory Effects of Pu-erh Tea Leaves on Mouse Type IV Allergy

Pu-erh tea is one of post fermented tea manufactured through oxidative maturing by microorganisms. We investigated the preventive effects of pu-erh tea extract on oxazolone-induced type IV allergy in male (ICR) mice. Oral administration of 50 mg/kg water extract of pu-erh tea leaves resulted in significant preventive effects against mouse type IV allergy. The hydrophilic Theabrownin-like fraction (TBW-ND), with a high molecular weight of approximately 12,000 or higher, was prepared from pu-erh tea leaves by a solvent-extraction method, followed by Diaion HP-20 column chro-matography and dialysis. Oral and percutaneous administrations of TBW-ND at doses of 18.7 mg/kg and 0.037 mg/ear also resulted in significant preventive effects, which included preventing increases in levels of the proinflam-matory cytokine interleukin-12. The anti-allergic ingredients in TBW-ND were predicted to be highly complex compounds containing of polyphenols, polysaccharides and/or proteins. The results suggest pu-erh tea leaves could be a beneficial food for protecting against delayed-type allergy.

Analysis of antioxidant prenylflavonoids in different parts of Macaranga tanarius,the plant origin of Okinawan propolis

Objective:To analyze the antioxidant prenylilavonoids in different parts of Macaranga tanarius(M.tanarius)(Euphorbiaceae)including the leaf,petiole,stem,leaflet,flower and fruit(only in female plant),and to evaluate their antioxidant properties.Methods:Methanol extracts of each part of M.tanarius were prepared and five prenylilavonoids in them were quantitatively analyzed using HPLC.The fruits from female plant were further separated into seed,pericarp,and glandular trichome.After the quantitative analyses of prenylflavonoids in each part of M.tanarius,antioxidant activity of the extracts was evaluated by 2,2-diphenyl-l-picryI-hydrazyl(DPPH)radical scavenging assay.Results:The leaf of M.tanarius contained two prenylflavonoids as main components in both male and female plants.Both flowers(male and female)contained five kinds of prenylflavonoids.In the petiole,stem and leaflet of both male and female plants,the prenylflavonoids were not delected or their amounts were very low.Five kinds of prenylflavonoids were detected in the seed,pericarp and glandular trichome of female M.tanarius.In particular,the glandular trichome had the highest level of total prenylflavonoids(235 mg/g of fresh plant).DPPH radical scavenging activity of all parts was more than 30%.Conclusions:We found that different parts of M.tanarius contained antioxidant prenylflavonoids.In particular,not only the glandular trichome but also the leaf contained prenylflavonoids,which indicated that M.tanarius may be developed as a functional plant,because the leaves of this plant can be easily collected.

Outcomes of Integrated Community Care Interventions for Frail Elderly People: A Literature Review

This study aimed to summarize the outcome measures and the significant outcome of effective integrated community care for frail elderly people through a literature review. A literature search was conducted using the Cochrane Library and PubMed for articles published up to November 2016 with the following search terms: Integrated community care, primary care, community, frail elderly, and effectiveness. A total of 106 articles were identified, of which eight with an interventional research design the inclusion criteria. All outcome measures were classified into the three categories: Functional abilities, quality of life, and health. As the significant effect, the physical function was perceived in four references, the quality of life in one reference, and the mental health in one reference. The evidence of the effectiveness of integrated community care seemed to be lacking. More studies will need to be conducted.

Improving Flow Property of Nifedipine Loaded Solid-Lipid Nanoparticles by Means of Silica for Oral Solid Dosage Form

In this study, a new formulation of silica nanocomposite containing nifedipine (NI) loaded freeze-dried solid-lipid nanoparticles (NI-SLNs) and silica have been developed with improved flowability of powders, which can lead to the formulation of a widely acceptable oral dosage form. The stable NI-SLNs were prepared using two phospholipids, hydrogenated soybean phosphatidylcholine and dipalmitoylphosphatidylglycerol mixed with 2.5% w/v trehalose as a cryoprotectant followed by lyophilization. We employed various grades of two types of silica, such as fumed and precipitated. Silica improved the poor flow property of NI-SLNs to good category as per USP-29. In addition, most of the silica nanocomposites showed the satisfactory results in their physicochemical properties such as particle size, polydispersity index, zeta potential, and recovered potency by around 100 nm, 0.3, -50 mV, and 80%, respectively. Furthermore, it was found that NI-SLNs were easily released form nanocomposites within 30 min, therefore, suggesting an improvement of drug dissolutions. Among them, precipitated silica cooperated fairly in improving the powder characteristics as well as the physicochemical, morphological, and pharmaceutical properties.

Photosafety testing of dermally-applied chemicals based on photochemical and cassette-dosing pharmacokinetic data

Drug-induced phototoxicity is elicited by exposure of skin and/or eyes to topical or systemic administration of chemicals,followed by exposure to sunlight.For photosafety testing,a new screening strategy,employing in vitro photochemical/photobiological and in vivo cassette-dosing pharmacokinetic(PK)studies,was proposed[1,2].The screening system provided reliable phototoxic predictions of tested chemicals at least within the same chemical series;however,the applicability of the proposed strategy is not fully understood.

ROLE OF REACTIVE OXYGEN IN MOUSE EPIDERMAL TUMOR PROMOTION

Generation of reactive oxygen(RO),has been proposedas an important event in mouse epidermal tumor promo-tion by 12-O-tetradecanoylphorbol-13-acetate (TPA). We investigate the role of RO in the promotion ofneoplastic transformation of JB6 mouse epidermal cellsby TPA using a battery of RO scavengers:CuZn-superoxide dismutase (CuZn-SOD),catalase。

Development of self-micellizing solid dispersion system employing amphipathic copolymer for the improvement of dissolution and oral bioavailability of cyclosporine A

The main objective of the present study is to develop a selfmicellizing solid dispersion(SMSD)system of cyclosporine A(CsA)using an amphiphilic copolymer,poly[MPC-co-BMA](pMB)to improve the biopharmaceutical properties of CsA(Fig.1A).Unlike conventional carrier compounds,pMB would perform the bifunctional ability as both polymeric carrier of solid dispersion system and solubilizer derived from a high micellizing property,which could be considered beneficial for the production of highly water soluble formulation of poorly water soluble compound[1].Improvement in the aqueous solubility has been believed to be a key consideration for acquiring potent pharmacological effects of BCS class II drug like CsA.

Discrete Circular Distributions with Applications to Shared Orthologs of Paired Circular Genomes

For structural comparisons of paired prokaryotic genomes,an important topic in synthetic and evolutionary biology,the locations of shared orthologous genes(henceforth orthologs)are observed as binned data.This and other data,e.g.,wind directions recorded at monitoring sites and intensive care unit arrival times on the 24-hour clock,are counted in binned circular arcs,thus modeling them by discrete circular distributions(DCDs)is required.We propose a novel method to construct a DCD from a base continuous circular distribution(CCD).The probability mass function is defined to take the normalized values of the probability density function at some pre-fixed equidistant points on the circle.Five families of constructed DCDs which have normalizing constants in closed form are presented.Simulation studies show that DCDs outperform the corresponding CCDs in modeling grouped(discrete)circular data,and minimum chi-square estimation outperforms maximum likelihood estimation for parameters.We apply the constructed DCDs,invariant wrapped Poisson and wrapped discrete skew Laplace to compare the structures of paired bacterial genomes.Specifically,discrete four-parameter wrapped Cauchy(nonnegative trigonometric sums)distribution models multi-modal shared orthologs in Clostridium(Sulfolobus)better than the others considered,in terms of AIC and Freedman’s goodness-of-fit test.The result that different DCDs fit the shared orthologs is consistent with the fact they belong to two kingdoms.Nevertheless,these prokaryotes have a common favored site around 70°on the unit circle;this finding is important for building synthetic prokaryotic genomes in synthetic biology.These DCDs can also be applied to other binned circular data.

Effect of Electrochemical Treatment in a Lithium Chloride Solution on Field Emission from Carbon Nanotubes

Carbon nanotubes （CNTs） are electrochemically treated in a lithium chloride solution at a concentration 0. 1 mol/L The field emission properties of the CNTs are investigated at different temperatures before and after the electrochemical treatment. After treatment, the turn-on voltage to produce field emission current of 10μA decreases from 4.2kV to 2. 7kV and the field emission current increases distinctly, but the stability falls off. Based on the Fowler-Nordheim plot, the values of the work function for the CNTs are calculated, which reveals that work function decreases after the electrochemical treatment. These results are attributed to the decrease of the work function of the carbon nanotubes.

The Function of Roots of Tea Plant (<i>Camellia sinensis</i>) Cultured by a Novel Form of Hydroponics and Soil Acidification

A novel form of hydroponic culture was employed to explore the physiological function of roots of a tea plant (Camellia sinensis). The pH of the nutrient solution with an actively growing tea plant decreased during cultivation. Furthermore, no oxalic acid, tartaric acid, malic acid or citric acid, all possible factors in acidification, was detected in the nutrient solution of a growing plant. A proton pump inhibitor suppressed the acidification of the solution. Soil acidification might have been accelerated with a proton released from ammoniacal nitrogen preferentially for the growth, suggesting the specific mechanism of tea plant as a functional food.

An Analysis of the Impact of Charging for Plastic Shopping Bags to Retailers in Japan

In accordance with a revision of the Containers and Packaging Recycling Law, there is an accelerating trend towards charging for plastic shopping bags （PSBs） in Japan for the purpose of reducing their discharge. The objectives of this research were to gauge, by means of a questionnaire survey, the opinions of retailers with regard to charging for PSBs, and to consider future directions for reducing the quantity of waste discharged. A questionnaire survey on charging for PSBs was conducted for retail stores in Shizuoka City. The survey was carried out in two phases. The results of the first phase （August to October 2008） showed that supermarkets were generally positive, while conversely other retailer types were negative. The results of the second phase （November to December, 2010）, targeting only supermarkets that had introduced charging after the first survey, showed that there was no major confusion and the shift proceeded relatively smoothly. In addition, the number of customers did not decline on account of charging, and the ratio of shoppers bringing their own reusable ＂eco bags＂ jumped dramatically. On these accounts, it is evident that charging for PSBs is an effective means of reducing usage of PSBs. The authors view it as important for the relevant governmental agencies to work towards promoting charging at retailers and simultaneously repeatedly explaining to consumers the necessity for charging.

Evaluating the Effects of the Concentration of Large Scale Municipal Solid Waste Incineration Facilities Using Geographical Information System

In Japan, each municipality generally has its own municipal solid waste incineration facilities because of the principle of self-management in the respective territories. Typically, a small municipality often owns a small facility. However, a large facility which can run continuously at high temperature with stability would be preferable if the dioxin generation mechanism is to be considered. Accordingly, municipalities in contiguous areas should cooperate mutually by using a large facility. To evaluate the effect of the concentration of large facilities, the authors created a GIS （Geographic Information System） based database of combustible waste generation at town level in Shizuoka Prefecture. Focusing on large facilities in Shizuoka City and superannuated facilities in Shida area, the authors evaluated the effect of the concentration of large facilities in Shizuoka city on utilization rate of facilities, energy balance and CO2 emissions. Our results showed the amount of light oil consumption and CO2 emissions increased because the mileage distance of garbage collection trucks becomes longer. However, the utilization rates of facilities and the amount of energy recovery from waste are improved. From these standpoints, the authors conclude that the concentration of large facilities is better compared to a single municipality based facility.

One-Step Preparation of Poly-Lactic-Co-Glycolic-Acid Microparticles to Prevent the Initial Burst Release of Encapsulated Water-Soluble Proteins

An initial burst is often observed during the release of active pharmaceutical ingredients (APIs) from poly-lactic-coglycolic-acid (PLGA) microparticles (MPs) which have been prepared by the emulsion-solvent evaporation method. Herein, we describe the development of a simple one-step coating method that suppresses the initial burst release process. This new method involves coating the PLGA-MPs with PLGA, with the coating process being performed through the phase separation of PLGA on the surface of PLGA-MPs using the emulsion-solvent evaporation method. Bovine serum albumin (BSA) was encapsulated in the PLGA-MPs as a model API. The coated MPs were spherical in shape with no pores on their smooth surface, whereas the non-coated PLGA-MPs had porous surfaces. An in vitro release study showed that the residual levels of BSA in the coated and non-coated PLGA-MPs after 1 h were about 99% and 16% of the original loads, respectively. The one-step coating method therefore represents a useful method for preparing PLGA-MPs that do not give an initial burst release of proteinaceous APIs.

Maintenance time of sedative effects after an intravenous infusion of diazepam: A guide for endoscopy using diazepam

AIM: To examine whether the sedative effects assessed by psychomotor tests would depend on the cytochrome P450 (CYP ) 2C19 genotypes after an infusion regimen of diazepam commonly used forgastrointestinal endoscopy in Japan. METHODS: Fifteen healthy Japanese volunteers consisting of three different CYP2C19 genotype groups underwent a critical ? icker fusion test, an eye movement analysis and a postural sway test as a test for physical sedative effects, and a visual analog scale (VAS) symptom assessment method as a test for mental sedative effects during the 336 h period after the intravenous infusion of diazepam (5 mg). RESULTS: The physical sedative effects assessed by the critical flicker test continued for 1 h (t values of 5 min, 30 min and 60 min later: 4.35, 5.00 and 3.19, respectively) and those by the moving radial area of a postural sway test continued for 3 h (t values of 5 h, 30 h, 60 min and 3 h later: -4.05, -3.42, -2.17 and -2.58, respectively), which changed significantly compared with the baseline level before infusion (P < 0.05). On the other hand, the mental sedative effects by the VAS method improved within 1 h. The CYP2C19 genotype-dependent differences in the postinfusion sedative effects were not observed in any of the four psychomotor function tests. CONCLUSION: With the psychomotor tests, the objective sedative effects of diazepam continued for 1 h to 3 h irrespective of CYP2C19 genotype status and the subjective sedative symptoms improved within 1 h. Up to 3 h of clinical care appears to be required after the infusion of diazepam, although patients feel subjectively improved.

Evaluation of Pollution Sources of Zinc in Tokyo Bay Based on Zinc Isotope Ratio in Sediment Core

This study evaluated the sources of Zn pollution in Tokyo Bay, Japan, on the basis of δ66Zn in a sediment core. The Zn concentration in sediments in the 1980s-2000s was considerably higher than the background concentration, suggesting that there remain important sources of Zn in the bay. The δ66Zn (+0.51‰) of anthropogenic Zn estimated in the core was significantly higher than those (approximately –0.1‰ - +0.2‰) of treated water from sewage treatment plants and vehicle-related sources. A large number of electroplating plants are located in Tokyo areas. It is assumed that Zn in effluents from electroplating operations is isotopically heavier owing to a negative isotopic effect on Zn electroplating. This tends to support the hypothesis that river bottom sediments, which were contaminated with Zn in the untreated effluents from electroplating plants in the past, are the principal sources of anthropogenic Zn in Tokyo Bay in the 1980s-2000s.

Antihypertensive effects of foods

Hypertension is one of the major risk factors for arteriosclerosis,which leads to cardiovascular disease and stroke.Several clinical trials revealed that control of the blood pressure is useful to reduce the morbidity and mortality associated with these diseases.However,the protective efficacy against these complications still remains at less than 50% even if the high blood pressure is treated by current medical drugs.Healthy diets are expected to not only prevent but also treat lifestyle-related diseases.Improvement of the dietary life,including lowsalt diets,appropriate alcohol consumption,and calorie restriction,is important for the prevention of hypertension.In addition,green tea,which has been drunk on a daily basis in Japan and China since ancient times,possesses an antihypertensive effect,and it was revealed that its components with this effect are catechins.Many studies have been performed on the antihypertensive effects of foods.Therefore,functional foods and their ingredients,reported to possess antihypertensive effects in animal experiments and human clinical trials,are summarized in this review.Blood pressure might be controlled by improvement of the daily eating habits based on evidence regarding these functional foods,and a healthy longevity can be expected.

Verification of Drainage Sterilization System Uses Low-Voltage Pulsed Electric Field in a Prawn Farm

It is necessary to take measures against infectious diseases in the Southeast Asian prawn farming industry. In giant tiger prawn （Penaeus monodon） aquaculture, diseases caused by viruses such as white spot syndrome virus （WSSV） and bacteria such as Vibrio have become a menace. Appropriate treatments of prawn culture pond＇s waters are advocated for preventing and controlling pathogens. The purpose of this study was to conduct an antimicrobial water treatment using a low-voltage pulsed electric field sterilization. Here we prepared a mechanical low-voltage pulsed electric field system with copper wire coiling around a titanium ring. The viability of WSSV in seawater was examined by prawn infectivity experiments. We inoculated healthy prawns （approximately 10-15 g） with the WSSV master sample that passed 0-3 times through the system. WSSV infection in prawns decreased according to the number of passes through the system. Healthy prawns survived for〉 10 days without feeding, where as prawns inoculated with the WSSV master sample showed symptoms of white spot and died in about 4 days. Two-thirds of the prawns inoculated with the WSSV master sample that was passed once through the system developed WSSV symptoms and were polymerase chain reaction （PCR） positive. However, no symptoms were observed and prawns were PCR negative when the WSSV master sample was passed three times through the system. Based on these results, we purpose that a low-voltage pulsed electric field system will serve as an efficient pond drainage sterilization system and will replace conventional treatments using chemicals such as sodium hypochlorite.