User-friendly relative quantification procedure for gas chromatography/mass spectrometry-based plasma metabolome analysis.

RATIONALE: Recently, metabolome analysis has been applied to a variety of research fields, but differences between batches or facilities can cause discrepancies in the results of such analyses. To resolve these issues using comprehensive metabolome analysis, in which it is difficult to perform quantitative analyses of all detected metabolites, internal standard compounds are used to obtain relative metabolite levels. This study investigated gas chromatography/mass spectrometry-based plasma metabolome analysis methods that are superior to relative quantification using internal standard compounds. METHODS: In experiment I, four analyses were performed under different analytical conditions at one facility, and then the data from the four analyses were compared. In experiment II, the same samples were analyzed at three facilities, and then the data from the three facilities were compared. RESULTS: Regarding the relative values obtained through comparisons with the internal standard compound, differences in the analytical results were observed among the four analytical conditions in experiment I and among the three facilities in experiment II, and the differences observed among the three facilities (experiment II) were larger. When correction was performed using plasma as a quality control, which is the procedure suggested in this study, these differences were markedly ameliorated. CONCLUSION: The suggested procedure involves the analysis of a plasma standard as a quality control for each batch and the calculation of relative target plasma to quality-control plasma values for each metabolite. This is an easy and low-cost method and could be readily employed by researchers during comprehensive plasma metabolome analysis.

Effect of Spinal Sagittal Alignment in Sitting Posture on Swallowing Function in Healthy Adult Women: A Cross-Sectional Study.

Swallowing function is both directly and indirectly related to postures, such as head and cervical angle and body position. However, the effects of different sitting postures on oropharyngeal swallowing have not been investigated. This study aimed to investigate whether the change in thoracolumbar alignment affected the oropharyngeal swallowing. A total of 58 healthy adult women (mean age 22.2 ± 1.67 years) without dysphagia were enrolled in this cross-sectional study. Participants were positioned in three sitting postures: comfortable sitting (CS), thoracic upright sitting (TUS), and slump sitting (SS). In each sitting posture, the kyphosis index (using a flexicurve), head and cervical angles (using a digital camera), swallowing speed (100-ml water swallowing test), and oral and articulatory function [by maximum tongue pressure (MTP) and oral diadochokinesis (ODK)] were evaluated. SS showed the largest kyphosis index and was associated with a greater anterior translation of the head. Swallowing speed was significantly decreased in SS compared with CS (p = 0.002) and TUS (p = 0.020) and ODK was significantly decreased in SS compared with other postures, for both /ta/ (p = 0.004) and /ka/ (p < 0.001) syllables. Further, MTP tended to decrease in SS compared with TUS (p = 0.064). Our results suggest that changes in sitting posture with different thoracolumbar alignments affect swallowing speed and oral and articulatory function. Consequently, adjustments to reduce sitting postural kyphosis may improve swallowing speed and oral and articulatory function.

Efficacy of soft palatal augmentation prosthesis for oral functional rehabilitation in patients with dysarthria and dysphagia: a protocol for a randomised controlled trial.

INTRODUCTION: Palatal augmentation prosthesis (PAP) is used in patients with articulation and swallowing disorders caused by postoperative loss of tongue tissue due to tongue cancer, cerebrovascular disease sequelae and age-related hypofunction. We have previously reported a newly designed soft PAP fabricated using an thermoplastic material that is particularly appropriate for early intervention. However, the effect of soft PAP on oral function improvement remains to be elucidated. The aim of this study is to investigate whether soft PAP can improve dysarthria and dysphagia occurring as cerebrovascular disease sequelae. METHODS AND ANALYSIS: This prospective, randomised, controlled trial will compare the immediate and training effects of rehabilitation using soft PAP with those of rehabilitation without using it. Primary outcomes are the single-word intelligibility test score and pharyngeal transit time (PTT). Secondary outcomes are tongue function (evaluated based on maximum tongue pressure, repetitions of tongue pressure and endurance of tongue pressure), articulation function (evaluated based on speech intelligibility, oral diadochokinesis, Voice-Related Quality of Life (V-RQOL)) and swallowing function (evaluated using Eating Assessment Tool-10). The study results will help determine the efficacy of Soft PAP in improving functional outcomes of word intelligibility and PTT. We hypothesised that early rehabilitation using Soft PAP would more effectively improve articulation and swallowing function compared with conventional rehabilitation without using soft PAP. ETHICS AND DISSEMINATION: Ethical approval was obtained from the Okayama University Certified Review Board. The study findings will be published in an open access, peer-reviewed journal and presented at relevant conferences and research meetings. TRIAL REGISTRATION NUMBER: jRCTs062200054.

Effects of Tongue-Strengthening Self-Exercises in Healthy Older Adults: A Non-Randomized Controlled Trial.

Tongue-strengthening exercises (TSE) using a device have been proposed as an intervention for improving tongue strength and endurance. However, devices for TSE have been expensive and difficult to manipulate and are not commonly used in home or clinical settings. This study therefore aimed to investigate whether tongue-strengthening self-exercises (TSsE) using a tongue-strengthening self-exercise tool at home can improve tongue strength in healthy older adults. This study included 27 participants (exercise group, η = 16, 7 men, 9 women, median age 84.5 years; control group, n = 11, 2 men, 9 women, median age 79.0 years). Exercises in the exercise group consisted of pushing the anterior tongue against the hard palate 30 times, 3 times a day, 5 days a week, for 8 weeks using a tongue-strengthening self-exercise tool. This tool is available in five levels of hardness. The most suitable hardness of the tool for each participant was calculated based on 60% of maximum tongue pressure (MTP) during the first 2 weeks of the training period and 80% of MTP for the remainder of the training period, as assessed using a tongue pressure-measuring device. The exercise group showed a significant improvement of 4.1 kPa in MTP (an 11.53% increase) and 4.53 s in endurance of tongue pressure (ETP) (a 99.86% increase). Furthermore, adherence in the exercise group was 99.2%. In conclusion, performing TSsE for 8 weeks was effective for increasing MTP and ETP in healthy older adults. This indicates that TSsE may be useful in older individuals at home to prevent age-related tongue muscle weakness.

Effects of Tongue-Strengthening Exercise on the Geniohyoid Muscle in Young Healthy Adults.

The activities of the suprahyoid muscles have been reported to be induced by tongue muscle contraction. The purpose of this research was to investigate whether tongue-strengthening exercises using a device cause hypertrophy of the geniohyoid muscle in healthy adults. Seven healthy young adults (3 men and 4 women, 21.0 ± 1.3 years old) received 8-week tongue muscle training with the JMS Tongue Pressure Measuring Device. The participants were instructed to press the anterior tongue against the hard palate 30 times in each session, three sessions a day, and 3 days a week. The exercise intensity was set to 60% of maximum tongue pressure in the first week, and to 80% of maximum tongue pressure for the remaining period. The training effect was evaluated by measuring (1) maximum tongue pressure value with the JMS Tongue Pressure Measuring Device, and (2) the area at rest, shortening amount, and contraction ratio of the geniohyoid muscle using ultrasonic imaging. After the 8-week training program, the maximum tongue pressure increased significantly from 44.9 to 61.6 kPa. The area of the geniohyoid muscle at rest also increased significantly from 2.3 to 2.6 cm2 after the program. There were no significant differences in the shortening amounts and the contraction ratios of the geniohyoid muscle between the values before and after the program. The tongue-strengthening exercise was useful to increase the muscle power of the geniohyoid, as well as the tongue muscles, in healthy young adults.

Effects of anterior tongue strengthening exercises on posterior tongue strength in healthy young adults.

OBJECTIVE: The aim of the present study was to investigate whether anterior tongue muscle strengthening exercises can affect the strength of posterior tongue muscles. DESIGN: Eleven healthy subjects (20.6 ± 1.2 years) were included. The subjects exercised by pushing the anterior tongue to the palate 30 times, three times a day, 3 days a week for 8 weeks. The exercise intensity was set at 60% of maximum tongue pressure (MTP) in the first week and 80% of MTP for the remainder of training. After the completion of training, MTP measurements were continued every month for another 3 months to evaluate whether training effects were sustained. RESULTS: MTP was significantly increased after 8 weeks of training compared with before training. No significant differences were seen between MTP immediately after completion of training and MTP 1-3 months after completion of training. However, MTP was significantly higher 1-3 months after completion of training than before training. CONCLUSIONS: The present study showed significant increases in both anterior and posterior MTPs by anterior tongue muscle strengthening exercises. In the future, a database on tongue muscle strengthening exercises in elderly persons, patients with dysphagia, etc. will need to be generated, with the aim of preventing frailty.

Estimation of P-glycoprotein-mediated efflux in the oral absorption of P-gp substrate drugs from simultaneous analysis of drug dissolution and permeation.

The purpose of this study was to establish an in vitro system that evaluates the effects of P-glycoprotein (P-gp)-mediated efflux on the oral absorption of P-gp substrates. An in vitro system (dissolution/permeation system, D/P system) was developed that consisted of apical and basal chambers and a Caco-2 cell monolayer mounted between the chambers. Both sides of the monolayer were filled with physiological solution and were stirred at 200rpm. The dissolution in the apical medium and permeation to the basal medium were monitored for 2h after P-gp substrates were applied to the apical side of the system. When erythromycin existed in the apical medium, the permeations of fexofenadine and talinolol were significantly enhanced without change in their dissolution. The prediction of oral absorptions of fexofenadine and talinolol from in vitro data indicated that co-administration of erythromycin results in 2.1- and 1.9-fold higher oral absorptions, respectively. Moreover, the D/P system could estimate the effect of cremophor EL on the oral absorption of saquinavir. These estimations corresponded well to in vivo human observations. Our in vitro system is useful in assessment of the effect of P-gp-mediated efflux on in vivo oral absorption of P-gp substrates.

The possible role of hematopoietic cell kinase in the pathophysiology of COPD.

BACKGROUND: Hematopoietic cell kinase (Hck) is a myeloid cell-specific tyrosine kinase, which is known to induce neutrophil infiltration to the lungs. Although the overexpression of Hck causes emphysema-like histologic changes in mice, its expression and activity in patients with COPD are unclear. METHOD: The aim of this study was to clarify the expression and activity of Hck in neutrophils from COPD patients, and to investigate the association between the degree of Hck expression and the lung function parameters in COPD patients. Peripheral blood neutrophils were isolated from 22 patients with COPD and 9 healthy subjects (HSs). The protein levels of Hck and phosphorylated Hck were assessed, and the correlation with various background characteristics was evaluated. RESULTS: The Hck protein level was significantly higher in neutrophils from COPD patients compared with HSs (COPD patients, 1.094; HSs, 0.801; p < 0.05). A significant positive correlation was observed between the protein level of Hck and the surface expression of the integrin molecule CD-11b (r = 0.540; p < 0.01) or CXC chemokine receptor-1 (r = 0.432; p < 0.05). In contrast, there was no difference in the phosphorylation of the Hck protein between COPD patients and HSs. CONCLUSION: The Hck protein level in peripheral blood neutrophils was increased in COPD patients, suggesting that Hck might have an important role in the neutrophil function and play a key role in the pathophysiology of COPD.

Overexpression of CD-11b and CXCR1 on circulating neutrophils: its possible role in COPD.

BACKGROUND: It has been shown that the beta2-integrin molecule is up-regulated in circulating neutrophils in COPD subjects. However, little has been reported about the expression of the cell surface molecules in such patients and their relationship with pulmonary function. The aim of the present study was to investigate the surface expression of molecules in circulating neutrophils and to clarify their possible role in the airflow limitation of COPD. METHODS: The surface expression of Mac-1 cells (ie, CD-11b and CD-18 cells) and CXC chemokine receptor (CXCR) 1 and CXCR2 of circulating neutrophils obtained from COPD patients and healthy subjects (HSs) was measured by flow cytometry analysis. The serum levels of interleukin (IL)-8 were measured by enzyme-linked immunosorbent assay. RESULTS: Both CD-11b and CXCR1 expression were significantly higher in COPD patients than in HSs (mean [+/- SE] CD-11b concentration: HSs, 9.7 +/- 1.0; COPD patients, 14.2 +/- 1.8 [p < 0.05]; mean CXCR1 concentration: HSs, 9.6 +/- 0.5; COPD patients, 11.9 +/- 0.4 [p < 0.01]). Although aging was positively correlated with the expression of CXCR1 (r = 0.440; p < 0.01), none of the other background factors, including smoking and body mass index, showed a correlation with the expression of the molecules. Although serum IL-8 levels were higher in patients with COPD than in HSs, no significant correlation between serum IL-8 levels and the expression of any molecule was seen. The expression of CD-11b (r = -0.317) and CXCR1 (r = -0.383) showed a significant negative correlation with the severity of airflow limitation (both p < 0.05). CONCLUSIONS: The overexpression of CD-11b and CXCR1 in circulating neutrophils may be associated with the development of airflow limitation in COPD patients.

Varicella-zoster virus gH:gL contains a structure reactive with the anti-human gamma chain of IgG near the glycosylation site.

Varicella-zoster virus (VZV) glycoproteins were purified from infected cells using monoclonal antibodies and gH:gL was found to react with antibodies to the gamma chain of human IgG (h-IgG), whereas gE:gI and gB did not. When gH:gL was captured by concanavalin A, it lost reactivity with the anti-h-IgG gamma chain (anti-h-gamma-IgG). gH:gL reacted with anti-h-gamma-IgG in an ELISA assay and gave a K:(d) value of 2.16x10(-7) M in a BIAcore assay. The K:(d) value of the human monoclonal antibody to gH (TI-57) used for the purification of gH:gL was 4.45x10(-10) M. Virus pretreated with anti-h-IgG was five times more resistant to neutralization with TI-57. Although the nature of the binding was not clear, gH:gL bound to anti-h-gamma-IgG. If this interaction results from immunological similarity between gH:gL and h-IgG, it may cause immune evasion in the pathogenesis of VZV infection.