Single-molecule analysis of intracellular insulin granule behavior and its application to analyzing cytoskeletal dependence and pathophysiological implications

Introduction: Mobilization of intracellular insulin granules to the plasma membrane plays a crucial role in regulating insulin secretion. However, the regulatory mechanisms of this mobilization process have been poorly understood due to technical limitations. In this study, we propose a convenient approach for assessing intracellular insulin granule behavior based on single-molecule analysis of insulin granule membrane proteins labeled with Quantum dot fluorescent nanocrystals.Methods: This approach allows us to analyze intracellular insulin granule movement with subpixel accuracy at 33 fps. We tracked two insulin granule membrane proteins, phogrin and zinc transporter 8, fused to HaloTag in rat insulinoma INS-1 cells and, by evaluating the tracks with mean-square displacement, demonstrated the characteristic behavior of insulin granules.Results and discussion: Pharmacological perturbations of microtubules and F-actin affected insulin granule behavior on distinct modalities. Specifically, microtubule dynamics and F-actin positively and negatively regulate insulin granule behavior, respectively, presumably by modulating each different behavioral mode. Furthermore, we observed impaired insulin granule behavior and cytoskeletal architecture under chronic treatment of high concentrations of glucose and palmitate. Our approach provides detailed information regarding intracellular insulin granule mobilization and its pathophysiological implications. This study sheds new light on the regulatory mechanisms of intracellular insulin granule mobilization and has important implications for understanding the pathogenesis of diabetes

Oil-in-water emulsion lotion providing controlled release using 2-methacryloyloxyethyl phosphorylcholine n-butyl methacrylate copolymer as emulsifier

AbstractLotion is a useful vehicle for active ingredients used to treat skin disease because it can be applied to the scalp, can cover large areas of skin, and it is easy to spread due to low viscosity. An emulsion lotion (EL) containing 2-methacryloyloxyethyl phosphorylcholine n-butyl methacrylate copolymer (PMB) as an emulsifier that provides controlled-release was developed. Diphenhydramine (DPH) was used as a model drug. Formulation with 5% DPH, 5% soybean oil, and 4% PMB in water was emulsified using a high-pressure homogenizer. Polysorbate 80 (TO) was used instead of PMB for comparison. They were applied in vitro to Yucatan micropig intact or stripped skin at a practical dose (2μL/cm2). For stripped skin, penetration of DPH from 4% PMB EL was slower than that from 1% TO EL; results for intact skin were similar. The same phenomenon was observed with application to rabbit skin in vivo. When 4% PMB EL dried on the skin, it made a thin film matrix incorporating the oil phase, which controlled the release of DPH. The release rate could be controlled by the ratio of oil phase to PMB. The EL with PMB shows promise as a vehicle for long-acting treatment of skin diseases

Introduction of thickened food for patient with dysphagia after surgical treatment for tumor in oral cavity

口腔腫瘍術後患者は,程度に差はあるが,嚥下･摂食障害をきたしやすい｡その経口摂取開始初期の栄養について,患者が満足して,より快適に摂取でき,経口のみで栄養が満たされるような食事を工夫した｡そのためには,食べやすさを増すためにトロメリンでとろみをつけ, また,食べ残しがないように患者の嗜好を取り入れた｡そして,この嗜好を取り入れたとろみ食を, 口腔腫瘍術後患者一例に提供した｡その結果, とろみ食は,食べやすく,全量摂取でき,良好な栄養状態を維持し,口内の移植皮弁へのトラブルもなく有効であるという結果が得られた｡また,患者が意欲や自信を持つという精神面への効果も見られた｡Following an operation to remove a tumor in the oral cavity, a patient with dysphagia who had received IVH and intubation feeding for 21 days, was given food thickened with proceeded starch and dextrin (Tromelin). The patient acceptance was good and he could chew and swallow safely and easily than if given ordinary food. The psychological and emotional effects of such postoperational care are also discussed

Coping behaviors with the experience of outpatients with gynecological cancer during adjuvant chemotherapy

research repor

Gender and coronary artery bypass graft surgery

Background Traditional and non-traditional risk factors for atherosclerotic cardiovascular disease (ASCVD) are different between men and women. Gender-linked impact of epicardial adipose tissue volume (EATV) in patients undergoing coronary artery bypass grafting (CABG) remains unknown. Methods Gender-linked impact of EATV, abdominal fat distribution and other traditional ASCVD risk factors were compared in 172 patients (men: 115; women: 57) who underwent CABG or non-coronary valvular surgery (non-CABG). Results In men, EATV, EATV index (EATV/body surface area) and the markers of adiposity such as body mass index, waist circumference and visceral fat area were higher in the CABG group than in the non-CABG group. Traditional ASCVD risk factors were also prevalent in the CABG group. In women, EATV and EATV index were higher in the CABG group, but other adiposity markers were comparable between CABG and non-CABG groups. Multivariate logistic regression analysis showed that in men, CABG was determined by EATV Index and other ASCVD risk factors including hypertension, dyslipidemia, adiponectin, high sensitive C-reactive protein (hsCRP) and type 2 diabetes mellitus (Corrected R2 = 0.262, p < 0.0001), while in women, type 2 diabetes mellitus is a single strong predictor for CABG, excluding EATV Index (Corrected R2 = 0.266, p = 0.005). Conclusions Our study found that multiple risk factors, including epicardial adipose tissue volume and traditional ASCVD factors are determinants for CABG in men, but type 2 diabetes mellitus was the sole determinant in women. Gender-specific disparities in risk factors of CABG prompt us to evaluate new diagnostic and treatment strategies and to seek underlying mechanisms

The Japanese space gravitational wave antenna; DECIGO

DECi-hertz Interferometer Gravitational wave Observatory (DECIGO) is the future Japanese space gravitational wave antenna. DECIGO is expected to open a new window of observation for gravitational wave astronomy especially between 0.1 Hz and 10 Hz, revealing various mysteries of the universe such as dark energy, formation mechanism of supermassive black holes, and inflation of the universe. The pre-conceptual design of DECIGO consists of three drag-free spacecraft, whose relative displacements are measured by a differential Fabry– Perot Michelson interferometer. We plan to launch two missions, DECIGO pathfinder and pre- DECIGO first and finally DECIGO in 2024

Herpesvirus protein ICP27 switches PML isoform by altering mRNA splicing

Viruses use alternative splicing to produce a broad series of proteins from small genomes by utilizing the cellular splicing machinery. Since viruses use cellular RNA binding proteins for viral RNA processing, it is presumable that the splicing of cellular pre-mRNAs is affected by viral infection. Here, we showed that herpes simplex virus type 2 (HSV-2) modifies the expression of promyelocytic leukemia (PML) isoforms by altering pre-mRNA splicing. Using a newly developed virus-sensitive splicing reporter, we identified the viral protein ICP27 as an alternative splicing regulator of PML isoforms. ICP27 was found to bind preferentially to PML pre-mRNA and directly inhibit the removal of PML intron 7a in vitro. Moreover, we demonstrated that ICP27 functions as a splicing silencer at the 3′ splice site of the PML intron 7a. The switching of PML isoform from PML-II to PML-V as induced by ICP27 affected HSV-2 replication, suggesting that the viral protein modulates the splicing code of cellular pre-mRNA(s) governing virus propagation

Isolation of Native Arbuscular Mycorrhizal Fungi within Young Thalli of the Liverwort <i>Marchantia paleacea</i>

Arbuscular mycorrhizal fungi (AMF) are a group of soil microorganisms that establish symbioses with most land plant species. &#8220;Root trap culture&#8221; generally has been used for isolating a single regenerated spore in order to establish a monospecific, native AMF line. Roots may be co-colonized with multiple AMF species; however, only a small portion of AMF within roots sporulate, and do so only under certain conditions. In this study, we tested whether young thalli (&lt;2 mm) of the liverwort Marchantia paleacea harbour monospecific AMF, and can be used as a vegetative inoculant line. When M. paleacea gemmae were co-cultivated with roots obtained from the field, the young thalli were infected by AMF via rhizoids and formed arbuscules after 18 days post-sowing. Ribosomal DNA sequencing of the AMF-colonized thalli (mycothalli) revealed that they harboured phylogenetically diverse AMF; however, new gemmae sown around transplanted mycothalli showed evidence of colonization from phylogenetically uniform Rhizophagus species. Of note, mycothalli can also be used as an inoculum. These results suggest that the young thalli of M. paleacea can potentially isolate monospecific AMF from field soil in a spore-independent manner