Development of an Imaging System Using Fluorescent Labeling Substances Excited by Infrared Rays

Abstract: An indocyanine green (ICG) derivative (ICG N-hydroxy sulfosuc-cinimide ester; ICG-sulfo-OSu) has been developed as an antibody labeling substance suitable for vital immunohistochemical staining. However, an appropriate fluorescence imaging system for ICG-sulfo-OSu using infrared rays has not as yet been reported. Therefore, we developed such a system. The absorption maxima of ICG and ICG-sulfo-OSu in buffer solution are both at 795 nm, the excitation maxima of their fluorescence spectra in buffer solution at 768 nm, and their emission maxima at 807 nm. An imaging system using an excitation filter with transmission at 710-790 nm and a barrier filter with transmission at 810-920 nm was constructed, and fluorescent images of ICG-sulfo-OSu labeled anti-epithelial membrane antigen (EMA) antibody were obtained with this system. Thus, vital immunohistochemical staining of microcancers under infrared ray excitation should now be possible, by exciting an ICG-sulfo-OSu labeled antibody specific to the tumor cells with infrared rays, using this imaging system.     

A side reaction in solid phase synthesis Insertion of glycine residues into peptide chains via Nim→ Nα transfer

In solid-phase peptide synthesis using N^α-Boc-N^im-tosyl-histidine (Boc-His(Tos)), byproducts having extra Gly residues in the peptide chain were observed at a high rate. When a Boc-amino acid such as Asn was incorporated after assembly of Boc-His(Tos), the N^im-tosyl group was partially or fully cleaved by an activating agent, 1-hydroxybenzotriazole. In the successive coupling reactions, Boc-Gly was incorporated into the free N^im ring as well as the α-amino function, and the N^im-Gly was then transferred to the α-amino group of Gly of the peptide chain after removal of these Boc groups to give extra Gly residues at the position of Gly. This was observed in only the coupling reaction with Boc-Gly and could be circumvented using a more stable N^im protecting group for His, such as a dinitrophenyl group.     

中药三七质量评价半微量方法及在三七连作障碍样品分析中的应用

该研究旨在建立三七质量评价半微量方法并应用于三七连作障碍样品分析。采用100倍量水饱和正丁醇超声提取0.1 g三七样品,所建立的半微量提取方法相比于甲醇常量提取法对5个主要皂苷类成分及三七皂苷K和Fa提取效率高9.6%~20.6%,且重复性良好,RSD2.5%。LC-MS-IT-TOF技术指认指纹图谱16个皂苷类成分特征峰,包括8个人参二醇型皂苷和8个人参三醇型皂苷。利用HPLC指纹图谱相似度评价及对特征峰峰面积提取可进行三七的质量分析。应用所建立的方法对人工干预克服连作障碍三七样品质量评价的结果表明:指纹图谱相似度、三七皂苷Fa的相对含量及三七皂苷K与三七皂苷Fa的比值(N-K/Fa值)均可作为评价连作地三七质量恢复程度的指标,其中N-K/Fa值同样适用于不同生长年限三七样品的区分。正常(无连作栽培)组样品三七皂苷K与三七皂苷Fa的峰面积比值为0.45~1.33;连作组样品个头小于正常组,指纹图谱与正常组相似度均低于0.87,三七皂苷Fa的峰面积显著低于正常组,N-K/Fa值显著高于正常组,为2.35~4.74,与正常组三七样品质量存在差异。人工干预一组的样品与正常组相似度均高于0.87,且16个特征峰的峰面积和分布模式均与正常组无显著差异,N-K/Fa值为0.42~2.06。人工干预二组的样品除重量较小的样品外,其余均与正常组指纹图谱相似度较高,三七皂苷K显著高于正常组,N-K/Fa值为0.96~6.16。连作障碍经人工干预后三七的质量均有不同程度的恢复和提高,其中人工干预一组的三七质量恢复情况优于人工干预二组。该研究建立的三七质量评价半微量方法,采用指纹图谱及峰面积比值作为评价指标可避免含量测定的繁琐及昂贵,为连作障碍三七样品的质量分析提供了简便、快速和准确的评价方法,且可应用于三七不同年限等样品的质量评价。     

Calcium supplementation and thyroid hormone protect against gentamicin-induced inhibition of proximal tubular Na+, K+-ATPase activity and other renal functional changes

Gentamicin can cause proximal tubule necrosis. We have shown that inhibition of PT Na⁺, K⁺-ATPase activity is rapidly induced by gentamicin. We have now investigated whether manipulations known to attenuate the negative effects of gentamicin on renal excretory capacity, i.e. high calcium intake and L-thyroxine treatment, will also attenuate gentamicin-induced inhibition of Na⁺, K⁺-ATPase activity and ameliorated signs of proximal tubule damage. Rats were gentamicin- or vehicle-treated for 7 days. Sub-groups were given 4% calcium (Ca) supplements or L-thyroxine 20 μg 100 g^-1 body weight daily. Gentamicin significantly reduced the glomerular filtration rate and increased the urinary excretion of the proximal tubule lysosomal enzyme, N-acetyl-β-d -glucosaminidase. Gentamicin significantly reduced proximal tubule Na⁺, K⁺-ATPase activity, measured in single permeabilized proximal tubule segments. Sodium excretion was inversely correlated to proximal tubule Na⁺, K⁺-ATPase activity. Both calcium and L-thyroxine alleviated all gentamicin-induced side-effects on renal function as well as on proximal tubule Na⁺, K⁺-ATPase activity. Calcium and L-thyroxine had no significant effect on renal function. L-thyroxine, but not calcium, increased proximal tubule Na⁺, K⁺-ATPase activity in control rats. Renal cortical tissue gentamicin concentration was not influenced by calcium but was significantly lowered by L-thyroxine. Two procedures which, via different mechanisms, afford protection from gentamicin-induced changes in renal function also give protection from gentamicin-induced inhibition of Na⁺, K⁺-ATPase activity. This suggests that loss of integrity of the Na⁺, K⁺-ATPase enzyme contributes to gentamicin-induced nephrotoxicity.     

Hybrid Assistive Limb (HAL) Rehabilitation in Patients with Acute Hemorrhagic Stroke

The efficacy of hybrid assistive limb (HAL) rehabilitation in the acute phase of stroke remains unclear. The purpose of this study was to evaluate the outcomes of patients with acute intracranial hemorrhage (ICH) who were treated with or without HAL rehabilitation. Among 270 patients with acute ICH from 2009 to 2014, 91 patients with supratentorial ICH were included in this retrospective study. Of these, 14 patients (HAL group) received HAL rehabilitation at approximately 1 week after ICH occurrence, while the remaining 77 patients received usual rehabilitation without HAL (N-HAL group). We obtained various patient data from the hospitals where the patients were moved to for further rehabilitation. Statistical comparisons were performed for the characteristics of the ICH patients, and outcomes between the HAL and N-HAL groups. There were no differences in outcomes between the HAL and N-HAL groups. However, patients with right ICH in the HAL group exhibited a significant association with a functional independence measure (FIM) score of ≥ 110 compared with patients in the N-HAL group (HAL group: 81.8%, N-HAL group: 43.9%, P = 0.04). In patients with right ICH, HAL rehabilitation was associated with improved outcomes as evaluated by the FIM score. Thus, HAL rehabilitation may improve outcomes of acute ICH in appropriately selected patients.     

Masticatory jaw movement optimization after introduction of occlusal interference

Summary How ‘control’ characteristics of masticatory jaw movement, such as skilfulness of the movement, change after alteration in occlusion remains uncertain. For each of 10 healthy adults with good occlusion, an occlusal interference with artificial ‘tooth‐cusp’ was introduced to the crown of the upper molar tooth on the non‐working side of unilateral chewing. Mandibular incisor‐point movements were then recorded by a 3D tracking device. The introduction of the occlusal interference induced a remarkable increase in the normalized jerk‐cost (NJC), prolonged duration of the decelerative phase and lowered peak velocity for jaw closing movement during chewing. Overall, the NJC and velocity profile showed significant recoveries during the course of about 90 repetitive chewing cycles performed under the altered occlusal condition. These findings suggest that acute adaptive changes of jaw motion after introduction of occlusal interference might be characterized as the recovery process of movement skilfulness in terms of movement smoothness and velocity profile.     

Cost-benefit Analysis of Comprehensive Mental Health Prevention Programs in Japanese Workplaces: A Pilot Study

We examined the implementation of mental health prevention programs in Japanese workplaces and the costs and benefits. A cross-sectional survey targeting mental health program staff at 11 major companies was conducted. Questionnaires explored program implementation based on the guidelines of the Japanese Ministry of Health, Labor and Welfare. Labor, materials, outsourcing costs, overheads, employee mental discomfort, and absentee numbers, and work attendance were examined. Cost-benefit analyses were conducted from company perspectives assessing net benefits per employee and returns on investment. The surveyed companies employ an average of 1,169 workers. The implementation rate of the mental health prevention programs was 66% for primary, 51% for secondary, and 60% for tertiary programs. The program’s average cost was 12,608 yen per employee and the total benefit was 19,530 yen per employee. The net benefit per employee was 6,921 yen and the return on investment was in the range of 0.27–16.85. Seven of the 11 companies gained a net benefit from the mental health programs.