CTAR®system, a hypoxia-induced and targeted anti-aging and repair system on human skin.

OBJECTIVE: The effective protection of skin cells and timely repair of damaged ones are of great significance for the maintenance of human skin's normal functions. Researches on skin cells mainly focus on the functions of repair factors. Relatively few studies have been made to clarify how skin cells sense the environment, absorb and transport nutrients, and achieve self-healing. CTAR®system, a new system, is proposed and demonstrated against human skin aging. MATERIALS AND METHODS: This study is made based on the research outcomes of the 2013 and 2019 Nobel Prize in Physiology or Medicine: precise regulation of cellular vesicle transport networks, and cellular perception and responding mechanisms to oxygen. The technologies and methods derived from the award-winning theories were applied in the field of skin anti-aging. RESULTS: It has shown that skin aging is a result of a series of injuries that have not been adequately repaired and then accumulate. The latest research on anti-aging of skin including channel identification, vesicle transport, induced activation, and damage repair has spawned CTAR®system, a hypoxia-induced and targeted anti-aging and repair system. CONCLUSIONS: We determined that a considerable part of skin aging problems can be partially solved by taking full advantage of the vesicular trafficking system in skin and improving skin's metabolism levels and physiological functions under low oxygen levels with CTAR®system.

Source of Energetic Protons in the 2014 September 1 Sustained Gamma-ray Emission Event.

We report on the source of > 300 MeV protons during the SOL2014-09-01 sustained gamma-ray emission (SGRE) event based on multi-wavelength data from a wide array of space- and ground-based instruments. Based on the eruption geometry we provide concrete explanation for the spatially and temporally extended γ -ray emission from the eruption. We show that the associated flux rope is of low inclination (roughly oriented in the east-west direction), which enables the associated shock to extend to the frontside. We compare the centroid of the SGRE source with the location of the flux rope's leg to infer that the high-energy protons must be precipitating between the flux rope leg and the shock front. The durations of the SOL2014-09-01 SGRE event and the type II radio burst agree with the linear relationship between these parameters obtained for other SGRE events with duration ≥ 3 hrs . The fluence spectrum of the SEP event is very hard, indicating the presence of high-energy (GeV) particles in this event. This is further confirmed by the presence of an energetic coronal mass ejection with a speed > 2000 km s - 1 , similar to those in ground level enhancement (GLE) events. The type II radio burst had emission components from metric to kilometric wavelengths as in events associated with GLE events. All these factors indicate that the high-energy particles from the shock were in sufficient numbers needed for the production of γ -rays via neutral pion decay.

Long-term Solar Activity Studies using Microwave Imaging Observations and Prediction for Cycle 25.

We use microwave imaging observations from the Nobeyama Radioheliograph at 17 GHz for long-term studies of solar activity. In particular, we use the polar and low-latitude brightness temperatures as proxies to the polar magnetic field and the active-regions, respectively. We also use the location of prominence eruptions as a proxy to the filament locations as a function of time. We show that the polar microwave brightness temperature is highly correlated with the polar magnetic field strength and the fast solar wind speed. We also show that the polar microwave brightness at one cycle is correlated with the low latitude brightness with a lag of about half a solar cycle. We use this correlation to predict the strength of the solar cycle: the smoothed sunspot numbers in the southern and northern hemispheres can be predicted as 89 and 59, respectively. These values indicate that cycle 25 will not be too different from cycle 24 in its strength. We also combined the rush to the pole data from Nobeyama prominences with historical data going back to 1860 to study the north-south asymmetry of sign reversal at solar poles. We find that the reversal asymmetry has a quasi-periodicity of 3-5 cycles.

Basic research and clinical application of beta-tricalcium phosphate (ß-TCP).

The mechanism of bone substitute resorption involves two processes: solution-mediated and cell-mediated disintegration. In our previous animal studies, the main resorption process of beta-tricalcium phosphate (ß-TCP) was considered to be cell-mediated disintegration by TRAP-positive cells. Thus, osteoclast-mediated resorption of ß-TCP is important for enabling bone formation. We also report the results of treatment with ß-TCP graft in patients since 1989. Two to three weeks after implantation, resorption of ß-TCP occurred from the periphery, and then continued toward the center over time. Complete or nearly complete bone healing was achieved in most cases within a few years and was dependent upon the amount of implanted material, the patient's age, and the type of bone (cortical or cancellous). We have previously reported that an injectable complex of ß-TCP granules and collagen supplemented with rhFGF-2 enabled cortical bone regeneration of rabbit tibiae. Based on the experimental results, we applied this technique to the patients with femoral and humeral fractures in elderly patients, and obtained bone union.

Measurement and comparison of individual external doses of high-school students living in Japan, France, Poland and Belarus-the 'D-shuttle' project.

Twelve high schools in Japan (of which six are in Fukushima Prefecture), four in France, eight in Poland and two in Belarus cooperated in the measurement and comparison of individual external doses in 2014. In total 216 high-school students and teachers participated in the study. Each participant wore an electronic personal dosimeter 'D-shuttle' for two weeks, and kept a journal of his/her whereabouts and activities. The distributions of annual external doses estimated for each region overlap with each other, demonstrating that the personal external individual doses in locations where residence is currently allowed in Fukushima Prefecture and in Belarus are well within the range of estimated annual doses due to the terrestrial background radiation level of other regions/countries.

Specific increase in non-functional masseter bursts in subjects aware of tooth-clenching during wakefulness.

Previous studies have reported that subjective awareness of a tooth-clenching habit is associated with increased jaw motor activity (Rao SM, Glaros AG, J Dent Res. 1979;58:1872). The aim of this study was to test the hypothesis that subjects with clenching awareness exhibit different motor expressions specific to non-functional oromotor activity under laboratory conditions without psychological or sensory effects. Polygraphic and audio-video recordings were made for a 30-min period of silent reading by 33 subjects without oro-facial pain. Oro-facial behaviours (e.g. swallowing, lip movements) were scored according to the polygraphic and audio-video records and masseter bursts were quantitatively analysed. Subjective psychological/sensory measures were also recorded before and/or after the polygraphic recording using a visual analogue scale. The subjects were classified into two groups one with 15 subjects who were aware of having a tooth-clenching habit and another with 18 who were not aware of any such habit. There were no differences between the groups with respect to the number of functional oro-facial behaviours or subjective psychological/sensory measures. Masseter bursts unrelated to functional oro-facial behaviours occurred more frequently in subjects with awareness [median (range) = 23 (2-187) bursts] than in those without [9.0 (0-36); P = 0.01], while neither burst activity [12.3 (1.8-34.5) % of maximum voluntary clenching and 10.1 (6.5-25.1) %, respectively] nor duration [1.17 (0.2-2.2) s and 1.28 (0.3-4.1) s, respectively] differed between the groups. The occurrence of functional oro-facial behaviours or other body behaviours (e.g. limb and body movements) did not differ between the two groups. These findings suggest that the increased masseter activity in subjects with tooth-clenching awareness is characterized by a specific increase in non-functional masseter bursts.

Relationship between tooth contacts in the retruded contact position and mandibular positioning during retrusion.

We conducted a series of studies with the purpose to investigate the locations of tooth contacts in the retruded contact position (RCP) and to discuss their significance in the stomatognathic system. In the present study, the relationship between the locations of RCP contacts and mandibular positioning during retrusion was examined. Thirty dentists and clinical residents were selected as subjects. One specialist in prosthetic dentistry examined each subject for the location of the RCP contacts. The mandibular positioning during retrusion was measured using a mandibular movement analysis system with six degrees of freedom. Originally programmed software was developed. Five reference points were selected: the central lower incisor (point I), the first molars on both sides (points RM and LM) and the condyles on both sides (points RC and LC). Tooth contact was observed most frequently at the second molar, followed by the first premolar. Points I, RM and LM all moved in an inferior-posterior direction, whereas points RC and LC moved in various directions ranging from superior-posterior to inferior-posterior. When the subjects were divided into two groups according to the most anterior tooth of occlusion in the RCP, the condylar positioning tended to be more superior in the group with molar contact than that with premolar contact. These results suggest that the locations of RCP contacts could be an important factor in jaw guidance during retrusion.

Relationship of periodontal bacteria and Porphyromonas gingivalis fimA variations with phenytoin-induced gingival overgrowth.

OBJECTIVES: We investigated the relationship between phenytoin-induced gingival overgrowth (GO) and the harboring of periodontal bacteria. MATERIALS AND METHODS: Periodontal conditions and subgingival bacterial profiles were examined in 450 sites of 75 subjects. A polymerase chain reaction method was used to detect six bacterial species; Porphyromonas gingivalis (Pg), Actinobacillus actinomycetemcomitans (Aa), Tannerella forsythia, Treponema denticola (Td), Prevotella intermedia (Pi), and Prevotella nigrescens (Pn). Genetic variations of the Pg fimA gene were also examined. Bacterial occurrence was compared with the severity of GO, and alterations in the bacterial occurrence rate and quantities were monitored following periodontal treatment. RESULTS: The occurrences of Aa, Td, Pi, Pn, and Pg with type II fimA (type II Pg) were significantly associated with the severity of GO. Td occurrence was reduced in association with gingival improvement following ultrasonic scaling, however, no such relationship was observed with Aa, Pi, Pn, and Pg. In addition, Pg and Pi markedly persisted after treatment. Clinical improvement of the sites, following an Er:YAG laser treatment, significantly associated with quantitative reduction of Pg in improved sites, however, not that of Pi. CONCLUSION: Type II Pg and Td were each found to have a significant relationship with the development and deterioration of GO.

220-fs erbium-ytterbium:glass laser mode locked by a broadband low-loss silicon/germanium saturable absorber.

We demonstrate femtosecond performance of an ultrabroadband high-index-contrast saturable Bragg reflector consisting of a silicon/silicon dioxide/germanium structure that is fully compatible with CMOS processing. This device offers a reflectivity bandwidth of over 700 nm and subpicosecond recovery time of the saturable loss. It is used to achieve mode locking of an Er-Yb:glass laser centered at 1540 nm, generating 220-fs pulses, with what is to our knowledge the broadest output spectrum to date.

Methylation pattern of CDH13 gene in digestive tract cancers.

Recently, the loss of CDH13 (T-cadherin, H-cadherin) gene expression accompanied by CDH13 promoter methylation was identified in colon cancers. We examined CDH13 methylation in oesophageal and gastric carcinomas. Five of 37 oesophageal cancers (14%) and 23 of 66 gastric cancers (35%) demonstrated abnormal methylation of the CDH13 promoter. Abnormal methylation was frequently found in gastric cancers of patients at all clinical stages just as in E-cadherin, another of the cadherin family, suggesting that these cancers could be methylated at an early stage. These results suggested that CDH13 might play a variety of roles depending on the tissue type.

CDH13 promoter region is specifically methylated in poorly differentiated colorectal cancer.

It has recently become clear that CDH13 (H-cadherin, T-cadherin) expression is frequently silenced by aberrant methylation in colorectal cancers and adenomas. In this study, we investigated the methylation status of CDH13 gene and detected aberrant promoter methylation in 27 of 84 (32%) colorectal cancers. We then correlated the results with the clinicopathological features of affected patients. We found a significant difference in histology (P=0.0053) when we compared the CDH13 methylation of poorly differentiated colorectal cancers to that of differentiated ones. This result suggested that poorly differentiated colorectal cancers specifically exhibited CDH13 methylation, and since CDH13 might be responsible for selective cell recognition and adhesion, inactivation of CDH13 could lead to the formation of scattered carcinoma cells in these cancers.

Detection of mitochondrial DNA alterations in primary tumors and corresponding serum of colorectal cancer patients.

We previously examined colorectal cancer patients using mutation-specific mismatch ligation assay for genetic alterations in primary tumors and paired serum samples and proved that genetic alterations present in the tumors of cancer patients can be detected in the serum of those same patients. Recent evidence has proved that various cancers frequently have mutations in the D-loop region of mitochondrial DNA (mtDNA). Therefore, we thought that mutations in the mitochondrial genome might also become a genetic marker of colorectal cancer to detect tumor DNA in the serum of patients. We first sequenced the D-loop region of mtDNA in colorectal cancers. We then proceeded with a sensitive method, i.e., mismatch ligation assay to examine the possibility that mtDNA alterations can be found in the serum DNA. We analyzed the D-loop region of mtDNA in 77 primary colorectal cancers, 7 of which (9%) contained true somatic mutations in this region. We then examined whether mtDNA alterations can be found in the serum DNA using mismatch ligation assay. Of 7 alterations that were examined, 1 (14%) could be detected in the serum. This result suggested that the mtDNA alteration could also be used as a tumor marker to detect tumor DNA in the serum.

Pharmacodynamics induced by direct electric current for the treatment of 5-fluorouracil resistant tumor: an animal experiment.

The repeated use of chemotherapy to treat patients with colorectal carcinoma may be limited by the fact it creates resistance cells. However, we have observed a remarkable decrease in certain types of drug resistance in patients treated with direct electric current. Experimental studies were therefore performed in animals to determine the differences in pharmacodynamics between chemotherapy with and that without electric treatment. Tumors were created in BALB/c mice by intradermal injection of 0.25 ml 4 x 10(6) Colon 26 cells/ml in the abdomen. Seven days later the mice were divided into two groups: controls and those that underwent electric treatment. Direct electric current (1,000 V, 0.2-0.8 microA) was passed between a platinum electrode inserted intradermally and the earth during and for 1 h after a single intravenous injection of 5-fluorouracil (5-FU; 12.5 mg ml(-1) kg(-1)). Peripheral blood samples were collected before and 5, 10, 20, 30, 45, and 60 min after the injection of 5-FU. Concentrations of 5-FU in the sera and tissues were measured by HPLC. The intratumoral concentrations of 5-FU in the electric treatment group were higher than those in the controls (P<0.05, two-factor analysis of variance), but the serum concentrations were not statistically different between the two groups. Pharmacodynamic changes were thus observed as a result of electrostatic treatment during chemotherapy. This elevated 5-FU concentration in the tumor tissue is considered one of the reasons for the effective inhibition of 5-FU resistance in clinical cases.

Adenovirus-mediated transduction of Escherichia coli uracil phosphoribosyltransferase gene increases the sensitivity of esophageal cancer cells to 5-fluorouracil.

Esophageal cancer is associated with the poorest prognosis among the digestive tract cancers, and chemotherapy is the treatment of choice for many patients. In this study, we experimentally introduced an Escherichia coli-derived uracil phosphoribosyltransferase (UPRT) gene to cultured esophageal cancer cell lines to potentiate the antitumor effects of a representative anticancer drug, 5-fluorouacil (5-FU). UPRT is a pyrimidine salvage enzyme that catalyzes the synthesis of uridine monophosphate from uracil and PRPP. The UPRT gene was transduced into five cultured esophageal cancer cell lines, TE1, TE2, TE3, NUEC1, and T.T, using an adenovirus vector. It was confirmed that the sensitivities of all cultured cell lines to 5-FU were increased in vitro. Subsequently, the T.T line was subcutaneously inoculated into nude mice to induce tumors, after which 5-FU was administered intraperitoneally. When a UPRT gene-recombinant adenovirus vector was directly injected into the tumors, tumor proliferation was markedly inhibited compared with that in the group treated with 5-FU alone, suggesting potentiation of 5-FU sensitivity by UPRT gene transduction in vivo. Therefore, we potentiated the effects of commercially available anticancer drugs by gene transduction. Our method may prove useful as a new form of cancer gene therapy in the future.

Colorimetric fluoride ion sensing by boron-containing pi-electron systems.

The boron-containing pi-conjugated systems, including tri(9-anthryl)borane (1) and tris[(10-dimesitylboryl)-9-anthryl]borane (2), have been investigated as a new type of fluoride chemosensor. Upon complexation of 1 with a fluoride ion, a significant color change from orange to colorless was observed and, in the UV-visible absorption spectra, the characteristic band of 1 at 470 nm disappeared and new bands around 360-400 nm assignable to pi-pi transitions of the anthryl moieties were observed. This change can be rationalized as a result of the interruption of the pi-conjugation extended through the vacant p-orbital of the boron atom by the formation of the corresponding fluoroborate. The binding constant of compound 1 with the fluoride ion was quite high [(2.8 +/- 0.3) x 10(5) M(-1)], whereas 1 only showed small binding constants with AcO- and OH- of around 10(3) M(-1) and no sensitivity to other halide ions such as Cl-, Br-, and I-, thus demonstrating its selective sensing ability to the fluoride ion. In contrast to the monoboron system 1, compound 2 having four boron atoms showed multistage changes in the absorption spectra by the stepwise complexation with fluoride ions.

Expression of mouse osteocalcin transcripts, OG1 and OG2, is differently regulated in bone tissues and osteoblast cultures.

Osteocalcin is a noncollagenous protein that is abundant in mineralized bone matrix. Mice have a gene cluster of osteocalcin that consists of OG1, OG2, and ORG. We established a new method to directly analyze the expression levels of OG1, OG2, and ORG mRNAs relative to total osteocalcin mRNA. They were amplified as 371-bp fragments by reverse transcription-polymerase chain reaction (RT-PCR) at the same time using common primers, digested with ApaLI, and separated in a polyacrylamide gel. ApaLI digestion did not affect the mobility of the OG1-derived 371-bp fragment, whereas both 371-bp fragments, derived from OG2 and ORG, were digested into 350 bp. Total RNA prepared from mouse bone was then subjected to RT-PCR followed by ApaLI digestion. OG1 and OG2 mRNAs were found to be expressed at ratios of 80%-86% and 14%-20%, respectively, to the total osteocalcin mRNA in mouse bone. The ratios were almost constant in various bones in vivo, independent of the animal's genetic background, age, or gender, or different parts of bone. RT-PCR using specific primers revealed that mouse bone tissues strongly expressed osteocalcin mRNA derived from OG1 and OG2, but not ORG. In contrast, cells cultured in vitro showed different expression ratios of osteocalcin mRNA: 53%-65% for OG1 and 35%-47% for OG2 to the total osteocalcin mRNA in the osteoblast cell line and primary osteoblasts in culture even though they formed many mineralized bone nodules. Similar results were obtained in both KS483 osteoblasts and C2C12 myoblasts, when they were cultured with bone morphogenetic protein-2 (BMP-2) to induce osteocalcin mRNA. Taken together, these findings indicate that OG1 is the predominant transcript among the three osteocalcin genes in mouse bone in vivo. It is also suggested that the expression of OG1 and OG2 is regulated differently in bone tissues and osteoblast cultures.

Positive regulation of cell-cell and cell-substrate adhesion by protein kinase A.

Integrin receptor activation is an important regulatory mechanism for cell-substrate and cell-cell adhesion. In this study, we explore a signaling pathway activated by mAb 12G10, an antibody that can activate beta(1) integrins and induce integrin-mediated cell-cell and cell-substrate adhesion. We have found that the cAMP-dependent protein kinase (PKA) is required for both mAb 12G10-induced cell-cell and cell-substrate adhesion of HT-1080 cells. Binding of mAb 12G10 to beta(1) integrins stimulates an increase in intracellular cAMP levels and PKA activity, and a concomitant shift in the localization of the PKA type II regulatory subunits from the cytoplasm to areas where integrins expressing the 12G10 epitope are located. MAb 12G10-induced cell-cell adhesion was mimicked by a combination of clustering beta(1) integrins and elevating PKA activity with Sp-adenosine-3',5'-cyclic monophosphorothioate or forskolin. We also show that two processes required for HT-1080 cell-cell adhesion, integrin clustering and F-actin polymerization are both dependent on PKA. Taken together, our data suggest that PKA plays a key role in the signaling pathway, resulting from activation of beta(1) integrins, and that this enzyme may be required for upregulation of cell-substrate and cell-cell adhesion.

Prognostic significance of multidrug resistance protein in adult T-cell leukemia.

The response of adult T-cell leukemia (ATL) to chemotherapy is poor, and a major obstacle to successful treatment is intrinsic or acquired drug resistance. To determine the clinical significance of multidrug resistance protein (MRP) 1 in ATL, we studied MRP1 expression and its association with clinical outcome. The expression of MRP1 mRNA in leukemia cells from 48 ATL patients was studied by slot blot analysis. The expression level of MRP1 mRNA in chronic-type ATL was significantly higher than that in lymphoma-type ATL (P = 0.033). There was no correlation between MRP1 expression and age, gender, WBC count, LDH, hypercalcemia, blood urea nitrogen, or performance status. However, the expression of MRP1 mRNA correlated only with peripheral blood abnormal lymphocyte counts (P = 0.008). The transporting activity of MRP1 was assessed using membrane vesicles. Membrane vesicles prepared from ATL cells with high expression of MRP1 mRNA showed a higher ATP-dependent leukotriene C(4) uptake than did those with low expression of MRP1 mRNA. This uptake was almost completely inhibited by LTD(4) antagonists ONO-1078 and MK571. In acute- and lymphoma-type ATL, high expression of MRP1 mRNA at diagnosis correlated with shorter survival, and Cox regression analysis revealed that MRP1 expression was an independent prognostic factor. These findings suggest that functionally active MRP1 is expressed in some ATL cells and that it is involved in drug resistance and has a possible causal relationship with poor prognosis in ATL. Multidrug resistance-reversing agents, such as ONO-1078 and MK571, that directly interact and inhibit the transporting activity of MRP1 may be useful for treating ATL patients.

Molecular detection of p16 promoter methylation in the serum of patients with esophageal squamous cell carcinoma.

PURPOSE AND EXPERIMENTAL DESIGN: Recent evidence shows that the presence of promoter hypermethylation of tumor suppressor genes has been demonstrated in the serum DNA of patients with various cancers such as lung, liver, and head and neck cancer. We have examined promoter hypermethylation of the p16 gene in esophageal squamous cell carcinoma (SCC) using methylation-specific PCR to detect tumor DNA in the serum. RESULTS: Aberrant promoter methylation of the p16 gene was detected in 31 of 38 (82%) esophageal SCCs. Subsequently, we tested for promoter methylation in the paired serum DNA of 31 patients with a p16 alteration in the primary tumor. We found that 7 of these 31 (23%) patients had the same methylation changes in the serum DNA. CONCLUSIONS: This result indicates that promoter methylation present in the tumors of esophageal SCC patients can be detected in the serum of the same patient and that this approach can potentially be used for the screening and monitoring of the disease.