Dual-Stimuli-Responsive Probes for Detection of Ovarian Cancer Cells and Quantification of Both pH and Enzyme Activity

Many physiological processes involve multiple coordinated chemical and/or biological events. Therefore, it is considered urgent to develop dual-responsive probes for a more comprehensive understanding of the synergistic effects between multiple analytes in complex cellular environments. In this study, we developed a dual-responsive probe βgal-BP-PMB (β-galactosyl–3, 3′-dihydroxy-2, 2′-bipyridyl–p-methoxybenzyl), the photoluminescence of which can be activated by β-galactosidase (β-gal) and acidic conditions. The overexpression of β-gal is an important feature of senescent and ovarian cancer cells. Single-input activatable probes for detecting β-gal activity in ovarian cancer cells can induce a false positive response from senescent cells. Because the lysosomal pH in senescent cells is increased, probe βgal-BP-PMB can be specifically activated in ovarian cancer cells, but silenced in senescent cells. Probe βgal-BP-PMB has a small molecular size, high sensitivity towards targeted stimuli and unique ratiometric properties, thereby enabling the quantification of both pH and enzyme activity. Such dual-responsive probes could earn a unique place in the field of bioimaging, where multiple analytes should be accurately and simultaneously monitored

Amphiphilic γ-cyclodextrin–fullerene complexes with photodynamic activity

Amphiphilic γ-cyclodextrin–fullerene 2 : 1 complexes (CLFCH complexes) were prepared by high-speed vibration milling of lipophilic tail-grafted γ-cyclodextrin (γ-CD), hydrophilic tail-grafted γ-CD and fullerene C₆₀. The transamidation of γ-CD–fullerene complexes having two amino groups with lipophilic and hydrophilic activated esters also afforded amphiphilic CLFCH complexes. Self-assemblies consisting of amphiphilic CLFCH complexes efficiently generated singlet oxygen under photoirradiation. Under visible light irradiation conditions, CLFCH complexes bearing a vitamin E moiety as a lipophilic tail showed high photodynamic activity toward cancer cells

モンゴルコク カチクニュウ オヨビ ニュウセイヒンチュウ ノ サイキン ノ クローン ライブラリーホウ ニヨル カイセキ

Various animal milks and their dairy products like AIRAG (fermented horse milk) and yogurt are very popular among Mongolian people. The climate of Mongolia is very severe, and they preserve their health by the intake of above animal milks and dairy products. In this study, we explored the diversity of bacteria in Mongolian animal milks (Cow, Horse, Goat, and Camel), AIRAG, and Camel milk yogurt by the clone library method of their 16S rRNA genes. Firstly, we prepared the whole genomic DNA from four animal milks and two dairy products, and amplified each 16S rRNA genes by PCR. PCR products (about 1.4kbp) were cloned into pGEM-T vector, and analyzed DNA sequences of totally 79 clones from AIRAG, Camel milk yogurt, and four animal milks. It was revealed that homologous clones to Lactobacillus helveticus are dominant in the clone libraries of both dairy products, whereas the other clones to Lactococcus from AIRAG, and Acetobacter from camel milk yogurt. Furthermore, We found that homologous clones to Lactococcus are dominant in Mongolian cow and horse milks, whereas Leuconostoc in camel milk. Therefore, it might be useful information for screening the bioactive strains from milk products in Mongolia

A Novel System for Transcutaneous Application of Carbon Dioxide Causing an “Artificial Bohr Effect” in the Human Body

BACKGROUND: Carbon dioxide (CO(2)) therapy refers to the transcutaneous administration of CO(2) for therapeutic purposes. This effect has been explained by an increase in the pressure of O(2) in tissues known as the Bohr effect. However, there have been no reports investigating the oxygen dissociation of haemoglobin (Hb) during transcutaneous application of CO(2)in vivo. In this study, we investigate whether the Bohr effect is caused by transcutaneous application of CO2 in human living body. METHODS: We used a novel system for transcutaneous application of CO(2) using pure CO(2) gas, hydrogel, and a plastic adaptor. The validity of the CO(2) hydrogel was confirmed in vitro using a measuring device for transcutaneous CO(2) absorption using rat skin. Next, we measured the pH change in the human triceps surae muscle during transcutaneous application of CO(2) using phosphorus-31 magnetic resonance spectroscopy ((31)P-MRS) in vivo. In addition, oxy- and deoxy-Hb concentrations were measured with near-infrared spectroscopy in the human arm with occulted blood flow to investigate O2 dissociation from Hb caused by transcutaneous application of CO(2). RESULTS: The rat skin experiment showed that CO(2) hydrogel enhanced CO(2) gas permeation through the rat skin. The intracellular pH of the triceps surae muscle decreased significantly 10 min. after transcutaneous application of CO(2). The NIRS data show the oxy-Hb concentration decreased significantly 4 min. after CO(2) application, and deoxy-Hb concentration increased significantly 2 min. after CO(2) application in the CO(2)-applied group compared to the control group. Oxy-Hb concentration significantly decreased while deoxy-Hb concentration significantly increased after transcutaneous CO(2) application. CONCLUSIONS: Our novel transcutaneous CO(2) application facilitated an O(2) dissociation from Hb in the human body, thus providing evidence of the Bohr effect in vivo

activator-induced quencher-detachmentに基づいたバイオイメージングのための酵素応答性 turn-on型蛍光プローブの開発

付記する学位プログラム名: 充実した健康長寿社会を築く総合医療開発リーダー育成プログラム京都大学新制・課程博士博士(工学)甲第23915号工博第5002号新制||工||1781(附属図書館)京都大学大学院工学研究科物質エネルギー化学専攻(主査)教授 大江 浩一, 教授 近藤 輝幸, 教授 浜地 格学位規則第4条第1項該当Doctor of Philosophy (Engineering)Kyoto UniversityDGA

Novel Surgical Treatment for Refractory Heel Ulcers in Werner’s Syndrome

Patients with Werner’s syndrome frequently develop chronic leg ulcers that heal poorly. We present a patient who suffered from this rare syndrome and developed typical heel ulcers. Treatment of the ulcer is challenging, as flap options are limited over the lower third of the leg and skin grafting is not easy as there is a lack of healthy granulations. We successfully treated the ulcer with osteomyelitis by drilling the bone and applying an ultrathin split thickness skin graft with the thigh skin as the donor site

Steric Control in Activator-Induced Nucleophilic Quencher Detachment-Based Probes: High-Contrast Imaging of Aldehyde Dehydrogenase 1A1 in Cancer Stem Cells

Turn-on fluorescence probes can visualize the enzyme activity with high contrast. We have established a new turn-on mechanism, activator-induced nucleophilic quencher detachment (AiQd), and developed AiQd-based turn-on fluorescence probes for the detection of enzymes. Herein, we demonstrate that the precise steric control efficiently quenches the fluorescence of AiQd-based turn-on probes before the enzymatic transformation. Theoretical calculation appropriately predicted the ratio of the fluorescence-quenched closed-ring form of probes. βC5S-A, which has a sterically demanding methyl group at the β-position of a fluorescence-quenching nucleophilic mercapto group, showed a low background signal. βC5S-A responded to aldehyde dehydrogenase 1A1 (ALDH1A1) with high selectivity, thereby enabling high-contrast live imaging of cancer stem cells (signal-to-noise ratio >10). The ALDH1A1-responsiveness of βC5S-A was not significantly affected by amino acids and biological thiols, such as cysteine and glutathione

Differences in Circulating microRNAs between Grazing and Grain-Fed Wagyu Cattle Are Associated with Altered Expression of Intramuscular microRNA, the Potential Target PTEN, and Lipogenic Genes.

We aimed to understand the roles of miRNAs in the muscle tissue maturation and those of circulating microRNAs (c-miRNAs) in beef production of Japanese Black (JB) cattle (Wagyu), a breed with genetically background of superior intermuscular fat depot, by comparing different feeding conditions (indoor grain-feeding vs. grazing on pasture). The cattle at 18 months old were assigned to pasture feeding or conventional indoor grain feeding conditions for 5 months. Microarray analysis of c-miRNAs from the plasma extracellular vesicles led to the detection of a total of 202 bovine miRNAs in the plasma, including 15 miRNAs that differed between the feeding conditions. Validation of the microarray results by qPCR showed that the circulating miR-10b level in the grazing cattle was upregulated compared to that of the grain-fed cattle. In contrast, the levels of miR-17-5p, miR-19b, miR-29b, miR-30b-5p, miR-98, miR-142-5p, miR-301a, miR-374b, miR-425-5p, and miR-652 were lower in the grazing cattle than in the grain-fed cattle. Bioinformatic analysis indicated that the predicted target genes of those c-miRNAs were enriched in gene ontology terms associated with blood vessel morphogenesis, plasma membrane, focal adhesion, endocytosis, collagen, ECM-receptor interaction, and phosphorylation. In the grazing cattle, the elevation of miR-10b expression in the plasma was coincident with its elevation in the longissimus lumborum (LL) muscle. Expression of bovine-specific miR-2478, the most plasma-enriched miRNA, tended to be also upregulated in the muscle but not in the plasma. Furthermore, grazing caused the downregulated mRNA expression of predicted miR-10b and/or miR-2478 target genes, such as DNAJB2, PTEN, and SCD1. Thus, the feeding system used for JB cattle affected the c-miRNAs that could be indicators of grain feeding. Among these, miR-10b expression was especially associated with feeding-induced changes and with the expression of the potential target genes responsible for glucose homeostasis and intramuscular fat depot in the LL muscle of JB cattle