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Background & Aims: While skeletal muscle index (SMI) is the most widely used indicator of low muscle mass (or sarcopenia) in oncology, optimal cut-offs (or definitions) to better predict survival are not standardized., Methods: We compared five major definitions of SMI-based low muscle mass using an Asian patient cohort with gastrointestinal or genitourinary cancers. We analyzed 2015 patients with surgically-treated gastrointestinal (n = 1382) or genitourinary (n = 633) cancer with pre-surgical computed tomography images. We assessed the associations of clinical parameters, including low muscle mass by each definition, with cancer-specific survival (CSS) and overall survival (OS)., Results: During a median follow-up period of 61 months, 303 (15%) died of cancer, and 147 died of other causes. An Asian-based definition diagnosed 17.8% of patients as having low muscle mass, while the other Caucasian-based ones classified most (>70%) patients as such. All definitions significantly discriminated both CSS and OS between patients with low or normal muscle mass. Low muscle mass using any definition but one predicted a lower CSS on multivariate Cox regression analyses. All definitions were independent predictors of lower OS. The original multivariate model without incorporating low muscle mass had c-indices of 0.63 for CSS and 0.66 for OS, which increased to 0.64-0.67 for CSS and 0.67-0.70 for OS when low muscle mass was considered. The model with an Asian-based definition had the highest c-indices (0.67 for CSS and 0.70 for OS)., Conclusions: The Asian-specific definition had the best predictive ability for mortality in this Asian patient cohort., (Copyright © 2023 Elsevier Ltd and European Society for Clinical Nutrition and Metabolism. All rights reserved.)

Introduction: Low health literacy (HL) is associated with poor surgical outcomes in patients with abdominal tumors, despite enhanced recovery programs. However, the relationship between HL and postoperative outcomes, including health-related quality of life (HRQOL) and physical functions, has been unclear in patients with abdominal tumors receiving perioperative rehabilitation programs (PRPs). Our objective was to identify associations of HL with postoperative functional and general outcomes in patients with abdominal tumors undergoing surgery and PRPs, controlling for critical covariates., Materials and Methods: The prospective cohort study was conducted from April 2020 to December 2021 in a single acute care hospital in Akita City, Japan. Out of 112 consecutive older adults with abdominal tumors referred to PRPs before surgery for tumors, 101 participated. Three patients declined undergoing follow-up assessment, and thus 97 were analyzed. Demographic data, comorbidities, education, surgical procedure, frailty status, cognitive functions, physical activity, nutritional status, tumor sites, types, and stages were collected as covariates at baseline. All participants underwent PRPs. HL was measured using the Japanese version of the European Health Literacy Survey Questionnaire, a comprehensive HL measurement. Primary endpoints were changes in HRQOL, measured using the EuroQol 5-dimension 5-level (EQ-5D-5L) index and EuroQol-visual analog scale (EQ-VAS), and physical functions (chair-stand ability, usual gait speed, and grip strength) from before surgery to discharge. Secondary endpoints were 90-day readmissions, postoperative complications, and length of hospital stay. Multivariate regression with inverse probability weighting, using propensity scores, estimeted associations of HL with the outcomes., Results: Of the 97 patients (36 [37.1%] women, mean [standard deviation] age 74.4 [6.3] years), 42 (43.3%) reported low HL. At baseline, patients with low HL exhibited worse EQ-5D-5L scores (P = .001), EQ-VAS values (P = .03), chair-stand ability (P = .001), and gait speed (P = .03) than controls. Low HL was associated with a low risk of grip strength declines (risk ratio, 0.13; 95% confidence interval, 0.12-0.99) but not with any other outcome., Discussion: There appeared to be no apparent associations of HL with surgical outcomes in patients with abdominal tumors receiving PRPs. Thus, HL might not be a crucial predictor of surgical outcomes when combined with PRPs., Competing Interests: Declaration of Competing Interest None reported., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

In this study, the phytoremediation potential of tropical and subtropical arsenic (As) hyperaccumulating fern Pteris vittata in an As contaminated farmland field near an abandoned goldmine was investigated. The tested field is located in a subarctic area of northeast Japan. This study was aimed at decreasing the risk of As in the soil (water-soluble As) with nurturing the soil and respecting the plant life cycle for the sustainable phytoremediation for 8 years. The field was tilled and planted with new seedlings of the fern every spring and the grown fern was harvested every autumn. The biomass and As concentration in fronds, rhizomes and roots of the fern were analyzed separately after harvesting each year. The biomass of the fronds of P. vittata was significantly affected by the yearly change of the weather condition, but As concentration in fronds was kept at 100-150 mg/kg dry weight. The accumulated As in P. vittata was higher than that of As-hyperaccumulator fern Pteris cretica, the native fern in the field trial area. Harvested biomass of P. vittata per plant was also higher than that of P. cretica. More than 43.5 g As/154 m 2 (convertible to 2.82 kg of As per hectare) was removed from the farmland field by P. vittata phytoremediation at the end of the 8-year experiment. Because of the short-term plant growth period and soil tilling process, total As in soil did not show significant depletion. However, the water-soluble As in the surface and deeper soil, which is phytoavailable and easily taken in cultivated plants, decreased to 10 μg/L (Japan Environmental Quality Standard for water-soluble As in soil) by the 8-year phytoremediation using P. vittata. These research data elucidate that the tropical and subtropical As hyperaccumulating fern, P. vittata, is applicable for As phytoremediation in the subarctic climate area., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Objectives: We aimed to identify the inspiratory muscle strength thresholds below which exercise capacity is compromised in men with chronic obstructive pulmonary disease (COPD)., Methods: We measured the maximum static inspiratory mouth pressure (PImax) and the percentage of predicted values (%PImax) in 113 patients with COPD. Six-minute walk distance (6-MWD) was measured as an indicator of functional exercise capacity, and a 6-MWD of <350 m was defined as functional exercise intolerance. Thresholds were determined as values with high specificity (>0.90) and maximal sensitivity. Statistical significance was set at P < 0.01., Results: The data of 96 patients (74 ± 6 years old; forced expiratory volume in 1-s [FEV 1 ], 56.5 ± 26.2% predicted) were analyzed; three women and 14 participants with missing data were excluded. Multivariate logistic regression models identified significant associations of PImax (odds ratio at 99% confidence interval [CI]: 0.95 [0.92-0.98]) and %PImax (odds ratio at 99% CI: 0.97 [0.95-0.99]) with 6-MWD, after adjustments for height and FEV 1 . C-statistics showed that the area under the curves (99% CI) of PImax and %PImax were comparable (0.87 [0.77-0.96] and 0.83 [0.72-0.94]). The thresholds of PImax and %PImax were 45.1 cmH 2 O and 66%; PImax and %PImax also had moderate positive likelihood ratios of 4.44 and 5.00., Conclusions: Thresholds of inspiratory muscle strength in men with COPD could help clinicians evaluate whether their patient's inspiratory muscle strength is inadequate to achieve a 6-MWD of ≥350 m, and identify patients who should be targeted for inspiratory muscle training., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Monocytes are differentiated into macrophages. In this study, mitochondrial DNA copy number (mtDNAcn) levels and downstream events such as the expression of respiratory chain mRNAs were investigated during the phorbol 12-myristate 13-acetate (PMA)-induced differentiation of monocytes. Although PMA treatment increased mtDNAcn, the expression levels of mRNAs encoded in mtDNA were decreased. The levels of mitochondrial transcription factor A mRNA and protein were also decreased. The levels of coenzyme Q10 remained unchanged. These results imply that, although mtDNAcn is considered as a health marker, the levels of mtDNAcn may not always be consistent with the parameters of mitochondrial functions., (Copyright © 2021. Published by Elsevier Inc.)

Introduction: The purpose of this study is to evaluate whether anatomical variations of the cystic duct and accessory bile duct can be grasped by cystic duct three-dimensional (3D)-computed tomography (CT) using non-contrast CT and to examine the possibility of omitting magnetic resonance cholangiopancreatography (MRCP)., Methods: Of patients who underwent non-contrast abdominal CT between May and October 2019, those who underwent MRCP within 1 month before and afterwards were targeted. Seven assessors visually evaluated the cystic duct 3D-CT images on a 5-point scale. Average scores of ≥3 and <3 points were assigned as the good and poor groups, respectively. Regions of interest (ROIs) were placed inside the cystic duct and four places around it, and the CT values in those ROIs were measured. The CT value difference was calculated by subtracting the surrounding CT values from the CT value in the cystic duct and converting the result to an absolute value. The CT value difference was classified into good and poor groups, and statistical analysis was performed. Seven assessors evaluated anatomical variations of the cystic duct and the presence of the accessory bile duct. The results were compared with the MRCP interpretation results to calculate sensitivity, specificity, positive predictive value, and negative predictive value., Results: The average visual evaluation score was 3.8. The good and poor groups were comprised by 53 (85.5%) and 9 (14.5%) patients, respectively. The CT difference value averages were 54.7 and 15.9 for the good and poor groups, respectively, and the value was significantly higher in the good group (p = 0.001). The comparison results with MRCP were sensitivity=83.3%, specificity=78.0%, positive predictive value=47.6%, and negative predictive value=95.1%., Conclusion: Cystic duct 3D-CT using non-contrast CT is a useful technique for understanding anatomical variations of the cystic duct and accessory bile duct. Our method may reduce the number of MRCP sessions performed., (Copyright © 2021. Published by Elsevier Inc.)

Absolute pitch (AP) is known as the ability to recognize and label the pitch chroma of a given tone without external reference. The neural mechanism and its asymmetry of AP musicians remain unclear. We herein report a 41-year-old AP musician who developed a right putaminal hemorrhage. On a postoperative day 5, a fluid-attenuated inversion recovery image revealed the rest of the hematoma and edematous lesion at the right white matter between the Heschl's gyrus and other cortices. Diffusion tensor tractography with the region of interest at the Heschl's gyrus was performed. In the left hemisphere, the anterior part of the arcuate fiber and middle longitudinal fasciculus were observed. However, these connections were absent in the right hemisphere, but her AP ability was maintained. Our case suggested that the fibers from the right Heschl's gyrus to the right frontal lobe via the right ventral stream is not associated with AP., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Background: The survival impacts of the prognostic nutritional index (PNI) and sarcopenia have been separately investigated in patients with gastric carcinoma (GC), while the prognostic impact of the combination of them remains to be addressed., Methods: In total, 1166 GC patients undergoing radical gastrectomy were retrospectively reviewed. A new prognostic score (PNIS) was developed based on preoperative PNI and sarcopenia; patients with both low PNI (≤44.8) and sarcopenia were allocated a score of 2, and those with only one or neither of these abnormalities were assigned a score of 1 or 0, respectively., Results: A lower PNI was independently associated with sarcopenia (P = 0.007). There were 704 (60.4%), 356 (30.5%) and 106 (9.1%) patients in the PNIS 0, 1 and 2 groups, respectively. A higher PNIS was associated with advanced age (P < 0.001) and a higher incidence of postoperative complications (P = 0.01). Patients with PNIS 2 showed significantly poorer overall survival (OS) than those with PNIS 1 or 0 (5-year OS; 57.8% vs. 79.2% vs. 91.6%, P < 0.001). Multivariate Cox hazards analysis showed PNIS 2 to be a powerful predictor of poor OS (HR 5.73, P < 0.001) in patients with pStage I disease, while not being independently associated with OS in those with pStage II/III disease. Patients with PNIS 2 had a markedly higher prevalence of non-GC-related death than those with scores of 0-1., Conclusion: The scoring system combining PNI and sarcopenia is useful for predicting survival outcomes, especially non-GC-related death, in patients with early GC, a population with basically good oncological outcomes., Competing Interests: Declaration of competing interest The authors have no conflicts of interest to disclose., (Copyright © 2020 Elsevier Ltd, BASO ~ The Association for Cancer Surgery, and the European Society of Surgical Oncology. All rights reserved.)

Background: Patients with oesophageal squamous cell carcinoma (ESCC) generally have distinctive body compositions; being underweight is highly prevalent and sarcopenic obesity is rare. We investigated the survival impacts of body mass index (BMI) in elderly (≥65 years) and non-elderly patients undergoing surgery for ESCC., Methods: In total, 379 ESCC patients were retrospectively reviewed. Patients were divided into 3 groups according to BMI; low (<20), medium (20-25) and high (≥25). The skeletal muscle index (SMI) was calculated and its relationship with BMI was analysed. Univariate and multivariate Cox hazards models were applied to determine independent predictors of poor overall survival (OS) and cancer-specific survival (CSS)., Results: The low-, medium- and high-BMI groups included 102 (26.9%), 231 (60.9%) and 46 (12.1%) patients, respectively. High BMI with low SMI was rare (n = 6, 1.6%). Patients with low BMI had significantly poorer OS and CSS than those with high and medium BMI (OS; P < 0.001, CSS; P = 0.003). Notably, OS and CSS curves were well-demarcated by BMI (both P < 0.001) in elderly patients, while not being stratified according to BMI in non-elderly patients (OS; P = 0.08, CSS; P = 0.54). Multivariable analysis revealed low BMI, as well as pStage III disease and non-curative resection, to be independent predictors of poor OS (HR 2.73, P < 0.001) and poor CSS (HR 2.88, P < 0.001) in the elderly group., Conclusions: The survival and oncological impacts of low BMI were evident only in elderly patients with ESCC. Our findings highlight the age-dependent significance of BMI in patients with this tumour entity., Competing Interests: Declaration of competing interest The authors have no conflicts of interest to declare., (Copyright © 2020 Elsevier Ltd, BASO ~ The Association for Cancer Surgery, and the European Society of Surgical Oncology. All rights reserved.)

Background: Although both sarcopenia and systemic inflammation reportedly affect long-term outcomes of esophageal carcinoma (EC) patients, their reciprocal associations with survival outcomes have yet to be investigated. This study aimed to evaluate the survival impact of sarcopenia combined with the neutrophil-to-lymphocyte ratio (NLR) in EC patients undergoing esophagectomy., Methods: In total, 378 EC patients were retrospectively reviewed. The cutoff value for NLR was set at the NLR median of the cohort. Sarcopenia was determined based on decreased skeletal muscle index calculated from computed tomography obtained before surgery. Univariate and multivariate Cox hazards models were applied to determine independent predictors of poor overall survival and cancer-specific survival., Results: Sarcopenia was more common in the high-NLR group (2.57 or greater) than in the low-NLR group (less than 2.57; P = .01). In the high-NLR group, patients with sarcopenia had significantly poorer overall and cancer-specific survival than those without sarcopenia (P < .001). In contrast, there was no survival impact of sarcopenia in the low-NLR group. Patients with both high NLR and sarcopenia exhibited poor overall and cancer-specific survival (5-year overall survival = 44.4%, 5-year cancer-specific survival = 57.0%). Sarcopenia was independently associated with poor overall survival (hazard ratio = 1.95; P = .007) and poor cancer-specific survival (hazard ratio = 2.66; P = .002) as well as pathological stage III disease and noncurative resection in the high-NLR group., Conclusions: The survival and oncological impact of sarcopenia was noteworthy only when present with elevated NLR. The combination of 2 factors is rational for identifying EC patients likely to have poor survival outcomes., (Copyright © 2020 The Society of Thoracic Surgeons. Published by Elsevier Inc. All rights reserved.)

Objectives: The survival impact of the preoperative prognostic nutritional index (PNI) has been investigated in older patients with gastric carcinoma (GC), while that of the postoperative PNI has yet to be addressed. We evaluated the significance of PNI before and after surgery in older GC patients (≥75 years)., Materials and Methods: In total, 309 older GC patients undergoing radical gastrectomy between 2006 and 2016 were retrospectively reviewed. The PNI was evaluated before and at six months after gastrectomy. Patients were divided into low (<45) and high (≥45) PNI groups. The impact of low PNI on overall survival (OS), cancer-specific survival (CSS), and non-GC-related death were investigated., Results: Low PNI was present in 134 patients (43.4%) preoperatively and 121 (39.2%) postoperatively. Low pre-PNI was independently associated with poor overall survival (P < .001). Similarly, OS was significantly stratified by post-PNI (P < .001). The significant survival difference according to post-PNI was present only in pStage I disease (P < .001). Low post-PNI independently increased the risk of non-GC-related death in a multivariable analysis (P = .002). In contrast, CSS was not stratified by post-PNI (P = .45). In the high pre-PNI group, total gastrectomy and super-older age (≥80 years) independently increased the risk of low post-PNI, which was significantly associated with poor survival outcomes., Conclusions: Pre- and post-operative PNI are useful for predicting long-term outcomes in older patients with GC. Low postoperative PNI is a powerful determinant of mortality due to other diseases. Optimal perioperative management is required for those at high risk of malnutrition postoperatively., Competing Interests: Declaration of Competing Interest The authors have no conflicts of interest to declare., (Copyright © 2020 Elsevier Inc. All rights reserved.)

We designed an electrode that has the ability to sense a target cell. This new electrode is intended for use in cell recognition via electron-transfer and cross-linker peptide immobilization. Myelopeptide-4 (MP-4:FRPRIMTP) is a marrow-origin peptide that interacts with receptors of the human leukemia cell line (K562 cells), and allows their differentiation. The YYYYC electron-transfer peptide improves the electron-transfer accessibility from an electroactive compound to an electrode. Oligoalanine plays the role of a cross-linker that immobilizes a peptide series (Ac-FRPRIMTPYYYYCAAAAA) to collagen, which then allows it to be cast onto an electrode. Use of the electrode with a peptide increased the peak currents of [Fe(CN) 6 ] 4-/3- and also improved the reversibility of redox. These improvements are due to the interaction between [Fe(CN) 6 ] 4-/3- and the peptide. When electrochemical impedance spectroscopy (EIS) measurements were carried out using a collagen/peptide probe-immobilized electrode, the electron transfer resisitance was lower than that without the peptide. The detection of K562 cells was based on an increase in resistance, because MP-4 was bound to the receptors on the cell surface. The responses were linear and ranged in number from 27 to 2,000 cells/mLwith a detection limit of 8 cells/mL. Recoveries of 50 and 1,000 cells/mL in human serum were accomplished at rates of 98 and 101%, respectively. Consequently, the proposed procedure is a powerful new concept for cytosensing., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Dermokine is a chiefly skin-specific secreted glycoprotein localized in the upper epidermis, and its family consists of three splice variants in mice and five in humans. To investigate the pathophysiological impact of dermokine, we generated mice deficient for two (βγ) or all dermokine isoforms (αβγ). Both variants, especially dermokine αβγ-deficient mice exhibited scale and wrinkle formation resembling ichthyosis accompanied by transepidermal water imbalance at the neonatal stage. Several dermokine αβγ-deficient mice died by postnatal day 21 when reared under low humidity. Moreover, the cornified envelope was vulnerable, and skin barrier lipid ceramides were reduced in the epidermis of dermokine αβγ-deficient mice. cDNA microarray and quantitative reverse transcriptase-PCR assays of the epidermis revealed the upregulation of small proline-rich protein and late cornified envelope family members, as well as antimicrobial peptides in the dermokine αβγ-deficient mice. These barrier gene signatures were similar to that seen in psoriasis, whereas recent studies demonstrated that congenital ichthyosis has gene profiles resembling psoriasis. In line with these findings, adult dermokine αβγ-deficient mice exhibited aggravated phenotypes in psoriasis-like dermatitis models but not in allergic dermatitis models. Dermokine may play a regulatory role in inflammatory dyskeratotic diseases, such as congenital ichthyosis and psoriasis, in the crosstalk between barrier dysfunction and inflammation., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)

We used an electron-transfer carbohydrate-mimetic peptide (YYYYC) to construct an electrochemical cytosensing system. Magnetic beads were modified with either asialofetuin (ASF) or soybean agglutinin (SBA) to evaluate the effect on cell sensing. Because SBA binds to the galactose residue that exists at the terminals of the carbohydrate chains in ASF, the target protein was accumulated on the protein magnetic beads. SBA is an example of N-acetylgalactosamine- and galactose-binding proteins that readily combine with YYYYC. When the peptides and protein-immobilized beads competed for a target protein, the peak current of the peptides changed according to the concentration of the protein at the 10 -12  M level. Next, human myeloid leukemia cells (K562 cell) were measured using the peptide and the carbohydrate chains on the cell surface that recognize SBA. The electrode response was linear to the number of K562 cells and ranged from 1.0 × 10 2 to 5.0 × 10 3  cells mL -1 . In addition, detection of a human liver cancer cell (HepG2 cell) was carried out using interactions with the peptide, the ASF receptors in HepG2 cells, and the carbohydrate chains of ASF. The peak currents were proportional and ranged between 5.0 × 10 1 and 1.5 × 10 3  cells mL -1 . When the values estimated from an electrochemical process were compared with those obtained by ELISA, the results were within the acceptable range of measurement error., (Copyright © 2018 Elsevier B.V. All rights reserved.)

For use in the voltammetric sensing of galactose-dependent proteins, we modified magnetic beads with a peptide that had both electroactive- and molecular recognition properties. The peptide consisted of a YXY sequence and behaved as an electron-transfer carbohydrate-mimetic peptide that would combine with proteins. With this tool, the protein could be detected via a label-free system. We synthesized several penta- and hexa-peptides with a cysteine residue on the C-terminals to examine the properties of peptides. These peptides contained amino acid residues (X) of alanine, serine, or tyrosine. The peptides were immobilized on magnetic beads via N-(8-maleimidocapryloxy) succinimide. Soybean agglutinin(SBA), the in vivo function of which has been well established in animals, was selected as a model protein. The protein was detected via the changes in electrode response due to the oxidation of tyrosine residues from the phenol group to quinone. As a result, SBA was selectively accumulated on the beads modified with YYYYC. The calibration curve of SBA was linear and ranged from 2.5 × 10 -12 to 1.0 × 10 -10  M. With this system, SBA was recovered in human serum at values that ranged from 98 to 103%. Furthermore, the beads with peptides were regenerated five times using a protein denaturant. Accordingly, this electrochemical system was simple and could be rapidly applied to the detection of galactose-recognition proteins., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Background: Sorafenib is a multi-kinase inhibitor for treating advanced hepatocellular and renal cell carcinomas by targeting various types of receptors and signaling molecules, including vascular endothelial growth factor receptors, platelet-derived growth factor receptor, and Raf-1. Sorafenib may cause diverse cutaneous adverse reactions, including hand-foot reaction, facial and scalp eruptions, alopecia and pruritus. However, the mechanism of these adverse effects has not been well-investigated., Objective: Mast cells (MCs) are reported to be associated with various types of skin diseases. To investigate the mechanism of sorafenib-induced cutaneous adverse effects, we focused on MCs in situ., Methods: We evaluated skin samples of organ cultured normal human skin treated with sorafenib using c-Kit, tryptase, and stem cell factor (SCF), Ki-67, and TUNEL immunohistochemistry as well as quantitative real-time polymerase chain reaction to evaluate MC number, degranulation, proliferation, and apoptosis in situ., Results: Sorafenib significantly increased the number and degranulation of skin-type MCs compared with the vehicle-treated control group in situ. However, sorafenib did not affect MC proliferation and apoptosis, suggesting that it stimulated MC maturation from resident precursors. Furthermore, sorafenib increased SCF expression in situ. The increase in MC number by sorafenib was abrogated by co-administration of SCF neutralizing antibody or the phosphoinositide 3-kinase (PI3K) inhibitor, wortmannin, but not the mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) kinase (MEK) inhibitor, PD98059. This suggests that SCF is involved in sorafenib-induced MC maturation. In addition, the compensatory upregulation of PI3K-signaling from inhibition of MAPK signaling by sorafenib might stimulate MC maturation in situ. We also evaluated MCs within the skin samples from patients with drug eruptions by sorafenib administration. The total and degranuated MCs number as well as SCF expression was significantly increased compared to healthy individuals., Conclusion: Our results contribute to a better understanding of the mechanism by which sorafenib induces adverse cutaneous reactions via activation of skin-type MC degranulation and maturation. This activation appears to be related to PI3K signaling and SCF production, which could be a new targets for treating sorafenib-induced adverse reactions., (Copyright © 2017 Japanese Society for Investigative Dermatology. Published by Elsevier B.V. All rights reserved.)

A carbohydrate/electro-transfer peptide probe was fabricated to perform cell sensing. The probe consisted of a cello-oligosaccharide that was created by the conjugation of an electron-transfer peptide (Y 5 C) and a carbohydrate via a Schiff base. An oxidation wave due to a phenolic hydroxyl group was obtained by scanning with a glassy carbon electrode. This cell-sensing system was based on a competitive reaction between carbohydrates on a cell surface and the probe as each reacted to a protein that recognized the carbohydrate. When amounts of the protein and probe were constant, the peak current of the probe was changed as the number of cells increased. A human histocytic lymphoma cell (U937 cell) with carbohydrates such as glucose and N-acetylglucosamine on its surface was selected as the target cell. Wheat germ agglutinin (WGA) binded to both the probe and the carbohydrates on U937 cells, which resulted in a linear peak current of the cellobiose/electron-transfer peptide at concentrations that ranged from 100 to 3500 cells/ml. The values of the cell sensing using this electrochemical method were consistent with those established via ELSIA. The sensitivity of this procedure, however, was two-fold superior to that of ELISA. Consequently, this carbohydrate/electron-transfer peptide could be a powerful tool for cell sensing and searching for carbohydrate chains on a cell surface., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Electrochemical sensing of ovalbumin (OVA) was performed using magnetic beads with OVA recognition (RNRCKGTDVQAW)/electron-transfer (YYYYC) peptides. The focus of this study was to construct a highly sensitive and regenerative tool for OVA detection based on the interaction between a protein and peptide-1(RNRCKGTDVQAWYYYYC). The peptide-1 was introduced to the bead through four types of cross-linking reagents. Magnetic beads of different sizes with N-(6-maleimidocaproyloxy)sulfosuccinimide (Sulfo-EMCS) were also prepared. An oxidation peak due to tyrosine residues at 0.65 V depended on the distance of the electron-transfer peptide from the bead surface and on the surface area of the magnetic beads that contacted the electrode surface. The response of the electro-transfer peptide moiety was decreased because the protein was accumulated via the recognition peptide on the beads. When using Sulfo-EMCS and beads that were 6.0-6.9 μm in diameter, the calibration curve of OVA was linear and ranged from 8.0 × 10 -13 to 2.0 × 10 -11  M. To regenerate the magnetic beads, the measurements were achieved after removal of the OVA using a denaturing reagent. When OVA was added to fetal bovine serum containing a complex matrix, OVA was recovered at a rate of 98-100%. Consequently, these magnetic beads could be a powerful tool for the sensing of OVA in real samples., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Lactate dehydrogenase (LDH) is a glycolytic enzyme that catalyzes the final step of glycolysis and produces NAD + . In somatic cells, LDH forms homotetramers and heterotetramers that are encoded by two different genes: LDHA (skeletal muscle type, M) and LDHB (heart type, H). Analysis of LDH isozymes is important for understanding the physiological role of homotetramers and heterotetramers and for optimizing inhibition of their enzymatic activity as it may result in distinct effects. Previously, we reported that hydroxychloroquine (HCQ) inhibited LDH activity, but we did not examine isozyme specificity. In the present study, we isolated heterotetrameric LDH (H 2 M 2 ) from swine brain, determined its kinetic and thermodynamic properties, and examined the effect of HCQ on its activity compared to homotetrameric LDH isozymes. We show that: (1) the K m values for H 2 M 2 -mediated catalysis of pyruvate or lactate were intermediate compared to those for the homotetrameric isozymes, M 4 and H 4 whereas the V max values were similar; (2) the K m and V max values for H 2 M 2 -mediated catalysis of NADH were not significantly different among LDH isozymes; (3) the values for activation energy and van't Hoff enthalpy changes for pyruvate reduction of H 2 M 2 were intermediate compared to those for the homotetrameric isozymes; (4) the temperature for half residual activity of H 2 M 2 was closer to that for M 4 than for H 4 . We also show that HCQ had different affinities for various LDH isozymes., (Copyright © 2016 Elsevier Inc. All rights reserved.)

To electrochemically sense lymphoma cells (U937), we fabricated a multifunctional peptide probe that consists of cell-penetrating/apoptosis-inducing/electron-transfer peptides. Electron-transfer peptides derive from cysteine residue combined with the C-terminals of four tyrosine residues (Y4). A peptide whereby Y4C is bound to the C-terminals of protegrin 1 (RGGRLCYCRRRFCVCVGR-NH2) is known to be an apoptosis-inducing agent against U937 cells, and is referred to as a peptide-1 probe. An oxidation response of the peptide-1 probe has been observed due to a phenolic hydroxyl group, and this response is decreased by the uptake of the peptide probe into the cells. To improve the cell membrane permeability against U937 cells, the RGGR at the N-terminals of the peptide-1 probe was replaced by RRRR (peptide-2 probe). In contrast, RNRCKGTDVQAWY4C (peptide-3 probe), which recognizes ovalbumin, was constructed as a control. Compared with the other probes, the change in the peak current of the peptide-2 probe was the greatest at low concentrations and occurred in a short amount of time. Therefore, the cell membrane permeability of the peptide-2 probe was increased based on the arginine residues and the apoptosis-inducing peptides. The peak current was linear and ranged from 100 to 1000 cells/ml. The relative standard deviation of 600 cells/ml was 5.0% (n = 5). Furthermore, the membrane permeability of the peptide probes was confirmed using fluorescent dye., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Hydroxychloroquine (HCQ) is a widely used drug in the treatment of autoimmune diseases, such as arthritis and systemic lupus erythematosus. It has also been prescribed for the treatment of malaria owing to its lower toxicity compared to its closely related compound chloroquine (CQ). However, the mechanisms of action of HCQ in erythrocytes (which bind preferentially this drug) have not been documented and the reasons underlying the lower side effects of HCQ compared to CQ remain unclear. Here we show that, although the activity of erythrocyte lactate dehydrogenase (LDH), but not GAPDH, was inhibited by both HCQ and CQ in vitro, LDH activity in erythrocytes incubated with 20 mM HCQ was not significantly reduced within 5 h in contrast to CQ did. Using HCQ coupled Sepharose chromatography (HCQ-Sepharose), we identified Band 3, spectrin, ankyrin, protein 4.1R and protein 4.2 as HCQ binding proteins in human erythrocyte plasma membrane. Recombinant cytoplasmic N-terminal 43 kDa domain of Band 3 bound to HCQ-Sepharose and was eluted with 40 mM (but not 20 mM) HCQ. Band 3 transport activity was reduced by only 23% in the presence of 20 mM HCQ. Taken together, these data demonstrate that HCQ binds to the cytoplasmic N-terminal domain of Band 3 in human erythrocytes but does not inhibit dramatically its transport activity. We hypothesize that the trapping of HCQ on Band 3 contributes to the lower side effects of the drug on energy production in erythrocytes., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Daunomycin penetrates the membrane of a U937 cell, which is a human histiocyte-related lymphoma cell. Several arginine-rich peptides have also exhibited a high degree of permeability with these cells. Therefore, we attempted to improve the membrane permeability of daunomycin by coupling it with an arginine-rich peptide. The cell membrane permeability of daunomycin was monitored using voltammetry, because daunomycin is an electroactive compound. First, daunomycin was combined with N-(6-maleimidocaproyloxy)sulfosuccinimide. Second, the cross-linking agent with daunomycin was bound to the cysteine residue of RRRRRRRRGC (peptide-1). The two-step synthesis suppressed the formation of by-products that might have conjugated with the amino groups of peptide-1. After the quinone moieties of daunomycin were reduced using an electrode, an oxidation peak appeared due to the moieties. The peak current of daunomycin with U937 cells had decreased. For the mixture of the daunomycin/peptide-1 probe and cells, the electrode response was smaller than that of daunomycin with the cells. Thus, the membrane penetration of the daunomycin/peptide-1 probe was improved compared with the use of only daunomycin. In addition, the membrane penetration of the probe was measured using fluorescence spectroscopy. The sensitivity of the electrochemical procedure was 100-fold that was obtained by fluorescence spectrometry., (Copyright © 2015 Elsevier B.V. All rights reserved.)

We designed a new electroactive peptide probe that has a molecular recognition function for the sensing of a protein. Ovalbumin (OVA) was the model protein, and when RNRCKGTDVQAW interacted with OVA, it conjugated with a tyrosine-rich peptide (Y4C). This peptide is electroactive, has a high degree of biocompatibility, and offers the possibility of gene expression. To measure the effect of a number of the tyrosine residues, voltammetric measurements were conducted using a series of tyrosine-rich peptides (YnC, n = 3-7) with sensitivities that ranged from 10(-9) to 10(-8) M. The electrode response of Y5C was the maximum value in the series. However, the peak current did not increase when the number of tyrosine residues was increased in a linear fashion. This may have been due to the micelles that are formed by a tyrosine-rich surfactant peptide. Thus, Y4C was suitable as an electroactive label for the construction of the peptide probe. The electrode response of Y4CRNRCKGTDVQAW obtained by a glassy carbon electrode was 100-fold that of tyrosine alone. The measurement of OVA via the peptide probe resulted in a detection on the order of 10(-12) M. In contrast, the sensitivity of OVA using RCKGTDVQAWY4C probe was at the 10(-11) M level, because the hydrophobic moiety gave it a molecular recognition function. The recoveries of the OVA using Y4CRNRCKGTDVQAW in a solution containing fetal bovine serum ranged between 98 and 101%. Consequently, the combination of a specific peptide and an electroactive element could be a powerful probe for the sensing of proteins., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Purpose: We evaluated the effects of low-intensity and home-based pulmonary rehabilitation (PR) on physical activity (PA) and the feedback provided by a pedometer in stable elderly patients with chronic obstructive pulmonary disease (COPD)., Methods: We assessed PA using a newly developed triaxial accelerometer (A-MES™, Kumamoto, Japan), which measures the time spent walking, standing, sitting and lying down. Twenty-seven elderly patients with COPD (age 74 ± 8 yrs; %FEV1 56.6 ± 18.7%) participated. They were randomly selected to undergo PR (pulmonary rehabilitation only) or PR + P (PR plus the feedback from using a pedometer). Their PA and pulmonary function, exercise capacity (6-min walking distance; 6MWD), quadriceps femoris muscle force (QF) were evaluated before the PR began (baseline) and at 1 year later. We compared the patients' changes in PA and other factors between the baseline values and those obtained 1 year later and analyzed the relationships between the changes in PA and other factors in the both groups., Results: The increase in the time spent walking in the PR + P group (51.3 ± 63.7 min/day) was significantly greater than that of PR group (12.3 ± 25.5 min/day) after the PR. The improvement rate of daily walking time after PR was significantly correlated with that of the 6MWD and QF in all subjects., Conclusions: These data suggest that low-intensity and home-based PR with the feedback from using pedometer was effective in improving PA, and the improvements of physiological factors were correlated with increased walking time in stable elderly patients with COPD., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

In this study, a peptide-1 (RNRCKGTDVQAW) constructing lysozyme was conjugated with an electroactive daunomycin in order to voltammetrically detect ovalbumin (OVA). Hetero-bifunctional cross-linking agents with four kinds of ethylene chains in differing lengths were used to bind the peptide-1 and daunomycin. After a cross-linking agent had reacted with an amino group of daunomycin, the compound was introduced into the peptide to the cysteine residue in the peptide using a pendant arm. The OVA was sensed via a change in the electrode response of the daunomycin moiety, based on the binding between the peptide and the OVA. The adsorption of the peptide probe on the electrode increased with increases in the ethylene chain. The binding constants between the peptide probes and the OVA, however, did not depend on the length of the chain. This was because the ethylene chain influenced the binding. When the peptide and the daunomycin were bound using N-(6-maleimidocaproyloxy) sulfosuccinimide, the electrode response of the peptide probe was the most sensitive from among the four cross-linking agents. The calibration curve of the OVA using the peptide probe was linear and ranged from 1.5×10(-11) to 3.0×10(-10)M. Furthermore, this method could be applied to the electrochemical sensing of the OVA in egg whites and in fetal bovine serum., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Objectives: The purpose of this study was to investigate the effect of anodal transcranial direct-current stimulation (tDCS) applied over the primary motor (M1) or the primary somatosensory (S1) cortices on somatosensory evoked magnetic fields (SEFs) following median nerve stimulation., Methods: Anodal tDCS was applied for 15min on the left motor or somatosensory cortices at 1mA. SEFs were recorded following right median nerve stimulation using a magnetoencephalography (MEG) system before and after the application of tDCS. SEFs was measured and compared before and after tDCS was applied over M1 or S1., Results: The source strengths for the P35m and P60m increased after tDCS was applied over M1 and that for the P60m increased after tDCS was applied over S1. The mean equivalent current dipole (ECD) location for the P35m was located significantly anterior to that of the N20m, but only during post 1 (10-20min after tDCS was applied over M1)., Conclusion: Our results indicated that the anodal tDCS applied over M1 affected the P35m and P60m sources on SEF components, while that applied over S1 influenced the P60m source., Significance: We demonstrated anodal tDCS applied over M1 or S1 can modulate somatosensory processing and components of SEFs, confirming the hypothesis for locally distinct generators of the P35m and P60m sources., (Copyright © 2014 International Federation of Clinical Neurophysiology. Published by Elsevier Ireland Ltd. All rights reserved.)

Background: The motor cortex in the human brain can be modulated by the application of transcranial static magnetic field stimulation (tSMS) through the scalp. However, the effect of tSMS on the excitability of the primary somatosensory cortex (S1) in humans has never been examined., Objective: This study was performed to investigate the possibility of non-invasive modulation of S1 excitability by the application of tSMS in healthy humans., Methods: tSMS and sham stimulation over the sensorimotor cortex were applied to 10 subjects for periods of 10 and 15 min. Somatosensory evoked potentials (SEPs) following right median nerve stimulation were recorded before and immediately after, 5 min after, and 10 min after tSMS from sites C3' and F3 of the international 10-20 system of electrode placement. In another session, SEPs were recorded from 6 of the 10 subjects every 3 min during 15 min of tSMS., Results: Amplitudes of the N20 component of SEPs at C3' significantly decreased immediately after 10 and 15 min of tSMS by up to 20%, returning to baseline by 10 min after intervention. tSMS applied while recording SEPs every 3 min and sham stimulation had no effect on SEP., Conclusions: tSMS is able to modulate cortical somatosensory processing in humans, and thus might be a useful tool for inducing plasticity in cortical somatosensory processing. Lack of change in the amplitude of SEPs with tSMS implies that use of peripheral nerve stimulation to cause SEPs antagonizes alteration of the function of membrane ion channels during exposure to static magnetic fields., (Copyright © 2014 Elsevier Inc. All rights reserved.)

For this study, a new method was developed to electrochemically detect ovalbumin via its binding with the peptide-1(RNRCKGTDVQAW) in lysozymes. The peptide that exists at the C-terminal of a lysozyme was combined with ovalbumin. When an electroactive compound was introduced to the N-terminal side of the peptide through ethylene gycolbis(sulfosuccinimidyl succinate), the labeled peptide-1 served as a probe for the detection of ovalbumin. The electrode responses of labeled peptide-1 were measured after the labeled peptide-1 and ovalbumin were incubated in a 0.1 M phosphate buffer (pH 5.6). As a result, the electrode response decreased as the concentration of ovalbumin increased. The detection limit of ovalbumin was 2.3 × 10(-11) M as estimated at 3-fold the standard deviation (3σ) (n = 5). Because the steric structure of the peptide and some of the amino acid residues were related to the binding, we prepared a peptide-2, to which the N- and C-terminals of peptide-1 were alternated. The decrease in the response for the labeled peptide-2 was less than that for the labeled peptide-1. In addition, the peak current of a peptide-3, for which the D of peptide-1 was replaced with S, was hardly changed with or without ovalbumin. Therefore, it was clear that the binding was influenced by the steric factors and by the sequence of the peptide. However, a peptide-1 with bis(sulfosuccinimidyl) suberate was designed to investigate the hydrophobic influences on the probe. The change in the peak current was smaller than that of peptide-1 with ethylene gycolbis(sulfosuccinimidyl succinate), which was due to the hydrophobic properties of the alkyl chain between the peptide and the ovalbumin. The proposed method could be applied to the determination of ovalbumin in egg whites. Consequently, the concept becomes an electrochemical sensing method for proteins based on the protein-peptide interaction., (Copyright © 2014 Elsevier B.V. All rights reserved.)

To electrochemically detect concanavalin A (ConA), a new method was developed using mixed micelles between a non-ionic surfactant with a maltose moiety and electroactive daunomycin. The surfactants, in which the length of the alkyl chain was different, were n-decyl-β-D-maltoside, n-dodecyl-β-D-maltoside, and n-tetradecyl-β-D-maltoside. The measurement principle was due to the micelle breakdown caused by the binding between the ConA and maltose moieties. When ConA was combined with maltose moieties at a concentration of surfactant that was near the critical micelle concentration, the daunomycin that formed the micelles was moved to a solution from the micelles. As a result, the peak current of daunomycin increased as the concentration of ConA was increased. The mechanism was proposed using voltammetry, spectrometry, and gel filtration. The linear range using n-tetradecyl-β-D-maltoside was 2.0×10(-9) to 8.0×10(-8) M of ConA, and it was the most sensitive in the presence of the three surfactants. To examine whether selective binding took place, measurements with several proteins were carried out. The electrode responses of daunomycin were not influenced by the presence of 5.0×10(-6) M protein. Furthermore, this method could be applied to the determination of ConA in a serum, and to the measurement of sugar chains that can be combined with ConA on the cell surface., (Copyright © 2014 Elsevier B.V. All rights reserved.)

P-cadherin serves as a major topobiological cue in mammalian epithelium. In human hair follicles (HFs), it is prominently expressed in the inner hair matrix that harbors the HF pigmentary unit. However, the role of P-cadherin in normal human pigmentation remains unknown. As patients with mutations in the gene that encodes P-cadherin show hypotrichosis and fair hair, we explored the hypothesis that P-cadherin may control HF pigmentation. When P-cadherin was silenced in melanogenically active organ-cultured human scalp HFs, this significantly reduced HF melanogenesis and tyrosinase activity as well as gene and/or protein expression of gp100, stem cell factor, c-Kit, and microphthalmia-associated transcription factor (MITF), both in situ and in isolated human HF melanocytes. Instead, epidermal pigmentation was unaffected by P-cadherin knockdown in organ-cultured human skin. In hair matrix keratinocytes, P-cadherin silencing reduced plasma membrane β-catenin, whereas glycogen synthase kinase 3 beta (GSK3β) and phospho-β-catenin expression were significantly upregulated. This suggests that P-cadherin-GSK3β/Wnt signaling is required for maintaining the expression of MITF to sustain intrafollicular melanogenesis. Thus, P-cadherin-mediated signaling is a melanocyte subtype-specific topobiological regulator of normal human pigmentation, possibly via GSK3β-mediated canonical Wnt signaling.

Objectives:  To develop effective electrical stimulation treatment to reduce spasticity, we examined the optimal stimulus point of the common peroneal nerve., Materials and Methods:  The locations of selective stimulus points for the deep peroneal nerve or superficial peroneal nerve fiber were examined in 25 healthy subjects in both legs (50 legs) using the ratio of the tibialis anterior (TA) to the peroneus longus (PL) M-wave amplitude (TA/PL ratio). In addition, we measured reciprocal Ia inhibition in ten healthy subjects. The amount of inhibition was determined from short-latency suppression of the soleus (Sol) H-reflex by conditioning stimuli to the deep or superficial peroneal nerve. The paired t-test was used for statistical analysis., Results:  The mean TA/PL ratio during deep peroneal nerve stimulation was significantly different from superficial peroneal nerve stimulation (p < 0.001). The mean stimulus point for the deep peroneal nerve was located 7 ± 5 mm distal and 3 ± 6 mm anterior from the distal edges of the head of fibula and was markedly different from the stimulus point for the superficial peroneal nerve (20 ± 7 mm distal and 12 ± 8 mm posterior). During deep peroneal nerve stimulation, the mean conditioned H-reflex was depressed to 83.8 ± 10.7% of the unconditioned value of the H-reflex. In contrast, during superficial peroneal nerve stimulation, the mean conditioned H-reflex increased to 105.3 ± 5.2%. These values were significantly different (p < 0.001)., Conclusions:  In the present study, we revealed a stimulus area of the deep peroneal nerve. Also, we observed the inhibitory effects of stimulation upon the deep peroneal nerve at individual stimulus point. Our results appear to indicate that localized stimulation of the deep peroneal nerve is more useful for the reduction of ankle spasticity., (© 2012 International Neuromodulation Society.)

Background: One of the major pathophysiologies in advanced chronic obstructive pulmonary disease (COPD) has been attributed to systemic inflammation. Meta-analysis of the 2005 Cochrane Database concluded the effect of nutritional supplementation alone on stable COPD was insufficient to promote body weight gain or exercise capacity. The aim of this study was to investigate the effectiveness of nutritional supplementation therapy using a nutritional supplement containing whey peptide with low-intensity exercise therapy in stable elderly patients with COPD., Method: In stable elderly COPD patients with %IBW and %FEV(1) of less than 110 and 80%, respectively, anti-inflammatory nutritional supplementation therapy was added to low-intensity exercise therapy. Thirty-six COPD patients were divided into those with and those without the ingestion of an anti-inflammatory nutritional supplement containing whey peptide, which exhibited an anti-inflammatory effect. These two groups were designated as the nutritional support and the control groups, respectively. The body composition, skeletal muscle strength, exercise tolerance, health-related QOL (HRQOL), and inflammatory cytokines were evaluated before and three months after nutritional support combined with exercise therapy in both the nutritional support group and the control group., Results: In the nutritional support group, the body weight, %IBW, FM, energy intake, %AC, Alb, PImax, PEmax, 6MWD, WBI, emotional function, and CRQ total were significantly increased, and the levels of hsCRP, IL-6, IL-8, and TNF-α were reduced significantly, while no significant change was noted in any item of physiological evaluation or any biomarker in the control group., Conclusion: Concomitant use of a anti-inflammatory nutritional supplement containing whey peptide, which exhibits an anti-inflammatory effect, with exercise therapy in stable elderly COPD patients with %IBW<110% and %FEV(1)<80% may not only increase body weight but may also inhibit systemic inflammation and thus improve exercise tolerance and HRQOL., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

To evaluate protein-protein interactions, a new voltammetric method was developed using a protein labeled with an electroactive compound. Concanavalin A (ConA), which is a lectin, recognizes α-mannose residues. Because the ConA was to be bound to ovalbumin (OVA), which has a high-mannose sugar chain, ConA labeled with daunomycin was prepared as the probe to monitor the binding. The binding to OVA was caused by the label modification of the ConA. As a result, the electrode response of the labeled ConA decreased as the OVA concentration increased. The electrode response of the labeled ConA was linearly over the range of 1.5×10(-10) and 1.5×10(-9)M OVA. The relative standard deviation of 1.5×10(-8)M labeled ConA and 1.5×10(-10)M OVA was 6.9% (n=5). The labeled ConA-OVA binding could then be conveniently monitored based on the change in response. In contrast, interactions between the labeled ConA and a protein with no specific sugar chain also were investigated. Incubation scarcely influenced the peak current of the labeled ConA. When several concentrations of OVA were added to a serum, good recovery determined it. Consequently, this method could be applied to the measurement of protein-protein interactions., (Copyright © 2011 Elsevier B.V. All rights reserved.)

We have recently shown that lipid mediators of the emerging endocannabinoid system (ECS) are key players of growth control of the human pilosebaceous unit. In this study, we asked whether the prototypic endocannabinoid anandamide (N-arachidonoylethanolamine, AEA) has a role in growth and survival of epidermal keratinocytes (KCs). Using human cultured KCs and skin organ-culture models, and by employing combined pharmacological and molecular approaches, we provide early evidence that AEA markedly suppresses KC proliferation and induces cell death, both in vitro and in situ. Moreover, we present that these cellular actions are mediated by a most probably constitutively active signaling mechanism that involves the activation of the metabotropic cannabinoid receptor CB(1) and a sequential engagement of the "ionotropic cannabinoid receptor" transient receptor potential vanilloid-1 (TRPV1). Finally, we demonstrate that the cellular effects of AEA are most probably due to a Ca(2+) influx via the non-selective, highly Ca(2+)-permeable ion channel TRPV1, and the concomitant elevation of intracellular Ca(2+) concentration. The data reported here may encourage one to explore whether the targeted manipulation of the above signaling pathway of the cutaneous ECS could become a useful adjunct treatment strategy for hyperproliferative human dermatoses such as psoriasis or KC-derived skin tumors.

Objective: To investigate the projection of muscle afferents to the sensorimotor cortex after voluntary finger movement by using magnetoencephalography (MEG)., Methods: The movement-evoked magnetic fields (MEFs) after voluntary index-finger extension were recorded by a 204-channel whole-head MEG system. Somatosensory-evoked magnetic fields (SEFs) were recorded after motor-point stimulation was applied to the right extensor indicis muscle by using a pair of wire electrodes., Results: The MEF waveforms were observed at 35.8±9.7 ms after movement onset (MEF1). The most concentrated SEFs were identified at 78.7±5.6 ms (M70), and the onset latency of M70 was 39.0±5.5 ms after motor-point stimulation. The mean locations of the equivalent current dipoles (ECDs) of MEF1 and M70 were significantly medial and superior to that of N20m elicited by median-nerve stimulation. The ECD locations and directions of both MEF1 and M70 were concordant in the axial, coronal and sagittal planes., Conclusions: MEF1 and M70 might be elicited by muscle-afferent feedback following muscle contraction. In addition, these ECDs may be located in area 4., Significance: Motor-point stimulation is a useful tool for confirming the projection of muscle-afferent feedback to the sensorimotor cortex after voluntary movement., (Copyright © 2010 International Federation of Clinical Neurophysiology. Published by Elsevier Ireland Ltd. All rights reserved.)

Study Objectives: The first aim of this study was to investigate the effects of nutritional supplementation combined with low-intensity exercise on body components, exercise tolerance, and health-related quality of life (HRQOL) in malnourished patients with COPD. The second aim of this study was to examine the degree of systemic inflammation and the actual changes in levels of systemic CRP, TNFα, IL-6 and IL-8 actual changes after a combination of nutritional supplementation and low-intensity exercise in these patients., Design: A prospective randomized trial., Patients: Thirty-two moderate to severe, clinically stable malnourished COPD patients., Methods: Patients were randomly divided into a nutritional supplementation with low-intensity exercise group and a control group. Lung function, maximum inspiratory and expiratory muscle force, the Chronic Respiratory Disease Questionnaire (CRQ), the 6-min walking distance (6MWD), and the Borg scale were measured at baseline and were re-assessed at 3 months after intervention. The degree of systemic inflammation and the changes in levels of systemic CRP, TNFα, IL-6 and IL-8 were assessed before and after a combination nutritional supplementation with low-intensity exercise., Results: Body weight and FFM increased significantly after 12 weeks of nutritional supplementation therapy in patients with COPD. The dietary intake energy increased and the REE:REEpred ratio decreased significantly in the nutrition with low-intensity exercise group. PI(max), Quadriceps muscle force and the 6-min walking distance (6MWD) increased significantly from baseline through week 12. Health status, as assessed by CRQ, improved in the domains of dyspnea and total sores significantly in the nutrition with low-intensity exercise group after intervention. In this group, hsCRP, IL-6, IL-8, and TNFα, decreased significantly after intervention compared with the control group., Conclusions: The combination of nutritional supplementation with low-intensity exercise training was successful in increasing weight and energy intake as well as exercise capacity and health-related QOL in our patients. Moreover, REE and major inflammatory cytokines decreased significantly after nutritional supplementation with low-intensity exercise training. The present study results suggest a potential role for the combination of nutritional supplementation and low-intensity exercise in the management of malnourished patients with COPD., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

To elucidate the basis of neuronal ceroid lipofuscinosis 1 (CLN1) from the viewpoint of enzyme structure, we constructed structural models of mutant palmitoyl protein thioesterase 1 (PPT1) proteins using molecular modeling software, jackal and TINKER. We classified the amino acid substitutions responsible for CLN1 and divided them into two groups, groups 1 and 2, based on the biochemical phenotype. Then, we examined the structural changes in the PPT1 protein for each group by calculating the solvent-accessible surface area (ASA) and the number of atoms affected. Our results revealed that the structural changes in group 1, which exhibits a complete deficiency of PPT1 activity, were generally large and located in the core region of the enzyme molecule. In group 2 exhibiting residual PPT1 activity, the structural changes in PPT1 were smaller and localized near the surface of the enzyme molecule. Coloring of affected atoms based on the distances between those in the wild type and mutants revealed the characteristic structural changes in the PPT1 protein geographically and semi-quantitatively. Structural investigation provides us with a deeper insight into the basis of CLN1., (Copyright 2009 Elsevier B.V. All rights reserved.)

Hair follicles (HFs) maintain a peripheral, functional equivalent of the hypothalamic-pituitary-adrenal (HPA) axis, whose most proximal element is corticotropin-releasing hormone (CRH). The mast cell (MC)-rich connective-tissue sheath (CTS) of mouse vibrissa HFs harbors MC precursors. Differentiation of these MC precursors into mature MCs can be induced by stem cell factor (SCF). We have investigated whether the MC progenitors of normal human scalp HF CTS respond to stimulation with CRH. Microdissected anagen HFs and full-thickness scalp skin were treated with CRH (10(-7) M). CRH treatment induced the degranulation of CTS MCs, in addition to increasing the number of CTS MCs in full-thickness skin and HF organ cultures in situ. In the latter, cells with characteristic MC features emigrated from the CTS. CRH-receptor protein expression in the CTS was colocalized with Kit expression on some CTS MCs in situ. CRH treatment upregulated SCF mRNA and protein expression within the HF epithelium. In skin organ culture, CRH-induced degranulation of CTS MCs was abolished by anti-SCF antibody. We demonstrate that human skin is an extramedullary reservoir for MC precursors, and we have identified a regulatory loop between CRH and SCF signaling. This highlights a previously unpublished finding about neuroendocrine control of human MC biology.

Background: We recently reported that the basement membrane (BM) zone components laminin-511 and -332 precisely regulate hair growth spatially and temporally during the anagen stage of the hair cycle., Objective: In this study, we examined the localization and roles of laminin-511 and -332 during catagen in mice., Methods: Using tissue from C57BL/6 hair depilation model mice, we performed immunohistochemistry, in situ hybridization, western blotting, and quantitative reverse transcriptase polymerase chain reaction (QRT-PCR) studies., Results: Although the distribution of laminin-332 around the BM of lower hair follicles changed during catagen, its total expression was stable throughout catagen stages at both the mRNA and protein levels. In sharp contrast, in situ hybridization, western blotting, and QRT-PCR studies of laminin alpha 5 showed that laminin-511 expression was gradually downregulated. Moreover, while the injection of recombinant laminin-332 at anagen stage VI did not affect catagen progression, injection of a laminin-511-rich A549 cell conditioned media protein extract at anagen stage VI delayed progression of catagen., Conclusion: These results indicated that downregulation of laminin-511 is important for hair regression., ((c) 2010 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All rights reserved.)