Search

Background Clostridium difficile (Cdf) releases toxins (TcdA and TcdB) that damage the intestinal epithelial barrier. Ecto-5′-nucleotidase ( CD73) is expressed on intestinal epithelial cells, and it is hypothesized to protect against toxin-induced epithelial damage through the cleavage of 5′- AMP to adenosine (Ado) and subsequent activation of adenosine receptors (AdoRs). Herein, we sought to assess the potential protective effects of CD73 and AdoR signaling on the injurious effects of Cdf toxins. Methods Barrier function was assessed with T84 colonocytes. Transepithelial electrical resistance ( TEER), paracellular fluorescein isothiocyanate ( FITC)-dextran flux, and tight junction protein ( ZO-1) integrity were monitored. Intrarectal installation of Cdf toxin was used to assess epithelial damage in vivo. Key Results TcdA/B caused reduced TEER and increased paracellular flux in vitro. Concurrent treatment with 5′- AMP attenuated these responses to Cdf toxin; an effect that was blocked with ZM241385 (AdoRA2 antagonist). APCP, a CD73 inhibitor, also suppressed the protective effects of 5′- AMP on paracellular flux. 5′- AMP reduced toxin-induced disruption of ZO-1, an effect that was abolished by APCP and ZM241385. Inhibition of CD73 with APCP during Cdf toxin exposure led to increased intestinal barrier permeability and epithelial damage in vivo. Intrarectal instillation of 5′- AMP had no effect on toxin-induced intestinal injury. Conclusions & Inferences Our data suggest that CD73 has a protective role against TcdA/B-induced damage. 5′- AMP treatment attenuated the damaging effects of Cdf toxin in vitro, and inhibitors of CD73 ( APCP) and AdoRs (ZM241385) revealed that the cleavage of 5′- AMP to Ado was necessary for the protective effects. Inhibition of CD73 in vivo increases colonic tissue damage and epithelial permeability during Cdf toxin exposure. [ABSTRACT FROM AUTHOR]

Background Cannabinoid type 1 (CB1) receptors are involved in the regulation of gastrointestinal (GI) motility and secretion. Our aim was to characterize the roles of the CB1 receptor on GI motility and secretion in vitro and in vivo by using different classes of CB1 receptor antagonists. Methods Immunohistochemistry was used to examine the localization of CB1 receptor in the mouse ileum and colon. Organ bath experiments on mouse ileum and in vivo motility testing comprising upper GI transit, colonic expulsion, and whole gut transit were performed to characterize the effects of the inverse agonist/antagonist AM251 and the neutral antagonist AM4113. As a marker of secretory function we measured short circuit current in vitro using Ussing chambers and stool fluid content in vivo in mouse colon. We also assessed colonic epithelial permeability in vitro using FITC-labeled inulin. Key Results In vivo, the inverse agonist AM251 increased upper GI transit and whole gut transit, but it had no effect on colonic expulsion. By contrast, the neutral antagonist AM4113 increased upper GI transit, but unexpectedly reduced both colonic expulsion and whole gut transit at high, but not lower doses. Conclusions & Inferences Cannabinoid type 1 receptors regulate small intestinal and colonic motility, but not GI secretion under physiological conditions. Cannabinoid type 1 inverse agonists and CB1 neutral antagonists have different effects on intestinal motility. The ability of the neutral antagonist not to affect whole gut transit may be important for the future development of CB1 receptor antagonists as therapeutic agents. [ABSTRACT FROM AUTHOR]

Background: A low level of serum magnesium ion (Mg 2+ ) is associated with type 2 diabetes mellitus (T2D). However, the molecular mechanism of Mg 2+ deficiency has not been fully clarified. The current study sought to assesses the effect of reactive oxygen species on the expression of Mg 2+ channels and miRNA., Methods: The expression of Mg 2+ channels and miRNA were examined by real-time polymerase chain reaction. Intracellular Mg 2+ concentration was measured by Magnesium Green fluorescence measurement., Results: The mRNA level of transient receptor potential melastatin 6 (TRPM6), which functions as Mg 2+ influx channel in the distal convoluted tubule (DCT) of the kidney, was decreased by glycated albumin (GA), but not by insulin in rat renal tubule-derived NRK-52E cells. The mRNA levels of TRPM7, a homologue of TRPM6, and CNNM2, a Mg 2+ efflux transporter located at the basolateral membrane of DCT, were changed by neither GA nor insulin. The generation of reactive oxygen species (ROS) was increased by GA. Hydrogen peroxide (H 2 O 2 ) dose-dependently decreased TRPM6 mRNA, but it inversely increased the reporter activity of TRPM6. H 2 O 2 accelerated the degradation of TRPM6 mRNA in actinomycin D assay without affecting TRPM7 and CNNM2 mRNA expressions. Nine miRNAs were considered as candidates for the regulator of stability of TRPM6 mRNA. Among them, miR-24-3p expression was increased by H 2 O 2 . The H 2 O 2 -induced reduction of TRPM6 mRNA was rescued by miR-24-3p siRNA. Magnesium Green fluorescence measurement showed that Mg 2+ influx is suppressed by H 2 O 2 , which was rescued by an antioxidant and miR-24-3p siRNA., Conclusions: We suggest that GA decreases TRPM6 expression mediated by the elevation of ROS and miR-24-3p in renal tubular epithelial cells of T2D.

Colonic epithelial cells move up along the crypt villus axis and are differentiated into absorptive or secretory cells. Claudin-7 (CLDN7), a tight junctional protein, is mainly located at the surface of crypt, whereas CLDN2 is located at the bottom. However, the expression mechanism and function of these CLDNs are not fully understood. The expression levels of CLDN2 and CLDN7 were altered depending on the culture days in MCE301 cells derived from mouse colon. The nuclear levels of transcriptional factors p53 and hepatocyte nuclear factor 4α (HNF4α) at day 21 were higher than those at day 7. Tenovin-1 (TEN), a p53 activator, increased the nuclear levels of p53 and HNF4α. The mRNA level and promoter activity of CLDN7 were increased by TEN, whereas those of CLDN2 were decreased. The changes of CLDNs expression were inhibited by p53 and HNF4α siRNAs. The association between p53 and HNF4α was elevated by TEN. In addition, the binding of p53 and HNF4α to the promoter region of CLDN2 and CLDN7 was enhanced by TEN. Transepithelial electrical resistance was decreased by TEN, but paracellular fluxes of lucifer yellow and dextran were not. In the Ussing chamber assay, TEN increased dilution potential and the ratio of permeability of Cl - to Na + . Both p53 and HNF4α were highly expressed at the surface of mouse colon crypt. We suggest that p53 and HNF4α alter the paracellular permeability of Cl - to Na + mediated by the inverse regulation of CLDN2 and CLDN7 expression in the colon.

We present a clinical case where a conservative treatment based on photonics [antimicrobial Photodynamic Therapy (aPDT) associated to Photobiomodulation therapy (PBMT)] of a patient with osteoradionecrosis (ORN) due to radiotherapy treatment of a laryngeal cancer. As a result of this combined treatment the ORN was controlled (e.g. the necrosis, infection and suppuration disappeared). Moreover, the symptoms reported by the patient (pain and xerostomia) also diminished along with the repair of oral mucosa. In future cases this combined therapy (e.g. PBM therapy and the aPDT) will be further examined., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Dietary NaCl depletion increases Na + and Cl - absorption in the colon, but the mechanisms are not fully understood. So far, we reported that the expression of claudin-7 (CLDN7), a tight junction (TJ) protein, was upregulated in the mice fed with NaCl-depleted diets, but the regulatory mechanism has not been clarified. Here, we found that angiotensin II (ANGII) increases the mRNA level of CLDN7, which was inhibited by losartan, a type 1 ANGII (AT1) receptor antagonist. Immunofluorescence measurement showed that CLDN7 is colocalized with zonula occludens-1 at the TJ in untreated and ANGII-treated cells. ANGII decreased transepithelial electrical resistance (TER) and increased permeability to C1 - without affecting permeability to lucifer yellow, a paracellular flux marker. In contrast, TER was increased by CLDN7 knockdown in the absence and presence of ANGII. ANGII increased the nuclear distribution of phosphorylated p65 subunit of NF-κB, which was inhibited by losartan. The ANGII-induced elevation of CLDN7 expression was blocked by BAY 11-7082 (BAY), an NF-κB inhibitor. Luciferase reporter assay showed that ANGII increases promoter activity of CLDN7, which was inhibited by the treatment with losartan or BAY, and introduction of mutations in κB-binding motifs in the promoter. The binding of p65 on the promoter region of CLDN7 was increased by ANGII, which was inhibited by losartan and BAY in chromatin immunoprecipitation assay. Our data suggest that ANGII acts on AT1 receptor and increases paracellular permeability to Cl - mediated by the elevation of CLDN7 expression in the colon.

Objectives: A standardized battery of quantitative sensory tests developed by the German Research Network on Neuropathic Pain (DFNS) was used to assess the association between somatosensory dysfunction and disease duration in patients with burning mouth syndrome (BMS)., Materials and Methods: The 28 female participants with BMS were classified according to disease duration: ≤ 6 months (subchronic BMS, n = 15) and > 6 months (chronic BMS, n = 13); 29 age- and sex-matched healthy volunteers (control group) were recruited from staff of a dental hospital. The DFNS quantitative sensory testing protocol was applied at the ulnar surface of the right forearm and the tip of the tongue. Values for BMS patients and controls were compared and analyzed., Results: The mechanical detection threshold (MDT) was significantly higher (i.e., loss of sensation) at the tongue tip in the chronic BMS group than in the control group (p = 0.011), whereas mechanical pain sensitivity (MPS) at the forearm was significantly higher (i.e., gain of sensation) in the chronic BMS group than in the control group (Z score = - 2.13 and 1.99, respectively). Multivariate analyses revealed that BMS patients could be discriminated from controls by using pressure pain threshold at the tongue (79.3%) (in the subchronic BMS group) and by MDT and MPS at the tongue tip and MPS at the forearm (96.6 and 89.7%, respectively) (in the chronic BMS group)., Conclusions: In BMS patients with long disease duration, MDT showed loss of sensation., Clinical Relevance: Increased MPS suggests that a neuropathic mechanism in the peripheral and central nervous systems is involved in BMS development.

Background: The simplistically and ambiguously termed 'fibrostenosis' of bowel is a hallmark of severe Crohn's disease [CD] and a major contributor to medical treatment failure. Non-invasive imaging assessment and novel medical therapy targeting this condition are under investigation, which particularly requires a better understanding of the underlying histological basis., Methods: We analysed 48 patients with stricturing Crohn's ileitis or/and colitis that required surgical resection. The most representative sections of the fibrostenotic, non-stenotic and uninvolved regions were reviewed for histological analysis. For each layer of bowel wall (mucosa including muscularis mucosae [MU], submucosa [SM], muscularis propria [MP], subserosal adventitia [SS]), histological abnormalities were evaluated individually, including active and chronic inflammation, fibrosis, smooth muscle hyperplasia or hypertrophy, neuronal hypertrophy and adipocyte proliferation. A novel semiquantitative histological grading scheme was created., Results: The most significant histopathological features characterizing the stricturing intestines were smooth muscle hyperplasia of SM, hypertrophy of MP and chronic inflammation. The muscular alteration was predominant in all layers. The overall muscular hyperplasia/hypertrophy was positively correlated with chronic inflammation and negatively correlated with fibrosis, whereas SM muscular hyperplasia was also associated with MU active inflammation. Similar changes, to a lesser extent, occurred in the adjacent non-stenotic inflamed bowel as well., Conclusions: In CD-associated 'fibrostenosis', it is the smooth muscle hyperplasia/hypertrophy that contributes most to the stricturing phenotype, whereas fibrosis is less significant. The 'inflammation-smooth muscle hyperplasia axis' may be the most important in the pathogenesis of Crohn's strictures., (Copyright © 2016 European Crohn’s and Colitis Organisation (ECCO). Published by Oxford University Press. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Background: Distinction between 2 forms of inflammatory bowel disease (IBD), ulcerative colitis (UC) and Crohn's disease (CD), can be challenging. Aberrant mucosal immunity suggests that CD is a T helper type 1 cell (Th1)-driven disease, whereas UC as Th2-driven response. However, whether this paradigm truly distinguishes CD from UC is controversial. We aimed to clarify the discriminating potential of lamina propria Th subsets in patients with IBD., Methods: Biopsies from 79 patients with IBD and 20 healthy controls were collected for Th subsets analysis (Th1:interferon γ [IFN-γ], T-bet; Th2:interleukin 13 [IL-13], Gata3; Th17:IL-17, RORγt; Treg:FoxP3). The receiver-operating characteristic curves were constructed to assess the discriminating ability by calculating the area under the receiver-operating characteristic curve. The equation with the highest area under the receiver-operating characteristic curve was applied to newly diagnosed patients to evaluate discriminating ability., Results: Patients with CD showed increased IFN-γ or T-bet cells and decreased IL-13 or Gata3 cells compared with UC. A discriminant equation composed of 4 markers (IFN-γ, T-bet, IL-13, and Gata3) yielded the highest area under the receiver-operating characteristic curve. In 36 established CD or UC, the sensitivity, specificity, positive and negative predictive probabilities were 92.6%, 55.6%, 86.2%, and 71.4% and in 14 newly diagnosed patients were 100.0%, 42.9%, 63.6%, and 100.0%. Furthermore, Gata3 cells were increased in tumor necrosis factor inhibitor therapy nonresponders compared with responders in CD. IFN-γ cells were directly and inversely proportional to disease activity in patients with CD and UC, respectively., Conclusions: The Th1/Th2 paradigm can distinguish CD from UC and may be further associated with response to tumor necrosis factor inhibitor in CD and disease activity in patients with IBD.

Background: IL-17 and Foxp3 double-expressing (DE) CD4(+) T lymphocytes are novel crossover immune cell population, but the presence and role of these cells in human intestinal inflammation is unclear. The aim of this study was to investigate the circulating IL-17 and Foxp3 DE CD4(+) T lymphocytes in patients with inflammatory bowel disease (IBD)., Methods: The entire cohort consisted of 79 subjects: 31 patients with Crohn's disease, 28 patients with ulcerative colitis, and 20 healthy control subjects (HC). IBD patients with evidence of active disease at endoscopy were entered into the study. Peripheral blood mononuclear cells were used for ex vivo and in vitro studies to assess the characteristics and generation of these novel cells and the function of circulating Foxp3 CD4(+) regulatory T lymphocytes (Treg) in patients with IBD compared with HC., Results: Patients with IBD had significantly higher prevalence of IL-17 and Foxp3 DE CD4(+) T lymphocytes compared with age- and gender-matched HC. These cells expressed RORγt. The ability of Treg cells to suppress autologous T-cell proliferation was reduced by approximately 60% in patients with IBD compared with HC. Increased generation of these DE cells was demonstrated by the modulation of cytokine environment of CD4(+) lymphocytes in vitro in patients with Crohn's disease., Conclusions: Prevalence of circulating IL-17 and Foxp3 DE CD4(+) T cells is increased in patients with IBD. Coexpression of RORγt and Foxp3 in these cells implies conversion from Treg cells to Th17 cells. This is associated with a decreased suppressive function of Foxp3 CD4(+) T lymphocytes in patients with IBD.

This study describes the association between low muscle mass and disability in performing IADL in elderly, community-dwelling Japanese subjects. Subjects were 1268 elderly, community-dwelling Japanese aged 65 years or older. Low muscle mass was defined by appendicular muscle mass index (AMI, weight [kg]/height [m(2)]), measured by bioelectrical impedance analysis (BIA). We classified "middle" AMI as a value 1-2 standard deviation (SD) below the young normal mean of AMI and "low" as more than 2 SD below the young normal mean. Examination of IADL was performed using the Tokyo Metropolitan Institute of Gerontology Index of Competence (TMIG-IC) questionnaire. In this study, 17.2% and 26.2% of elderly men and women, respectively, were classified as having low muscle mass. There was a significant association with age and the change from normal to middle to low muscle mass in both men and women. The prevalence of IADL disability also increased significantly as muscle mass decreased. After adjustment for age and BMI, low muscle mass was significantly associated with IADL disability in men and women. Furthermore, middle muscle mass was significantly associated with IADL disability in women. This study revealed that low muscle mass, defined as a value 2 SD below the mean of young adults, had a significant association with IADL disability in elderly Japanese. Interventions to prevent age-related losses in muscle mass may be an effective strategy to prevent IADL disability among the elderly., (Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.)

Objectives: Diagnostic criteria for the metabolic syndrome (Mets) in Japan have been set by the Medical Committee of the Japanese Association of Medical Sciences (Med), the National Health and Nutrition Examination Survey (Nat), specific health checkups (Ckup), and second medical examination by Worker's Accident Compensation Insurance System (Wor). The purpose of this study was to compare classification of the metabolic syndrome by different organizational criteria and to investigate underlying differences., Methods: All faculty members of a university in Osaka, Japan, underwent mandatory health checkups in September 2008. The demographic distribution included 769 males (mean age, 49 +/- 12 years) and 415 females (mean age, 43 +/- 10 years). Using the Med, Nat, Ckup and Wor criteria, individuals were assessed for the MetS and pre-metabolic syndrome (pre-Mets), strongly suspected metabolic syndrome (S-Mets) and assumed pre-metabolic syndrome (A-pre-Mets), as well as a positive support level (PSL) and a motivational support level (MSL). All faculty members were categorized into a morbid group (Mets, S-Mets, PSL, and FB) or a pre-morbid group (pre-Mets, A-pre-Mets, and MSL) based on medical data and smoking habits. The incidence of morbid and pre-morbid individuals was compared across the four criteria and analyzed based on gender and age (under 40 and 40 or over)., Results: Male incidences for the morbid and pre-morbid classifications were 17% and 20% with Med, 9% and 23% with Nat, 27% and 14% with Ckup, and 1.4% and 0% with Wor. There were significant differences across criteria sets in both the morbid and pre-morbid groups, with significantly greater numbers of males than females, and higher prevalences in those aged 40 or over than in their younger counterparts. Males aged under 40 classified into the pre-morbid group comprised 18% in Med, 16% in Nat, and 13% in Ckup., Conclusion: The different disease incidences found between Med and Ckup data in males aged 40 or over might be attributed to varying criteria for blood glucose levels, while Wor data may be influenced by the higher level of blood pressure set as a criterion with this approach. It will be important to continuously validate currently established criteria to identify the actual prevalence of MetS in Japan. Furthermore, incorporation of waist circumference and BMI for females, and a positive approach for young males, may be critical for future developments.

Aim: The aim of this study was to objectively assess the risk parameters associated with walking in housebound elderly., Methods: The subjects were 622 community-dwelling elderly (210 men and 375 women: age 65-85) with independence in activities of daily living (ADL). We administered questionnaires pertaining to housebound factors and measured walking parameters, twice, in May 2008 and May 2009. Housebound status was defined as leaving the house no more than once a week, and non-housebound status as more than once every 2-3 days. We measured the following walking parameters: usual and maximum walking speed, timed up and go, obstacle-negotiating gait, stair-climbing, and number of daily steps., Results: The overall prevalence of housebound status was 10.0% in men and 8.5% in women. No statistically significant differences were seen in the prevalence of housebound status between genders or age groups. In men, maximum walking speed, timed up and go, and daily steps were slower or less in housebound than in non-housebound subjects. In women, all walking parameters, except usual walking speed, were slower or less in housebound than in non-housebound subjects. Logistic regression analysis showed that obstacle-negotiating gait was a risk factor for men (odds ratio 2.49), and for women, all walking parameters, except walking at usual speed, were risk factors, with the highest odds ratio of 4.77 for obstacle-negotiating gait. A slower obstacle-negotiating gait was a risk factor for housebound status for both men and women., Conclusion: Compared with non-housebound subjects, housebound elderly with the ability to go out alone had a similar usual walking speed but a slower obstacle-negotiating gait.

The aim of this study is to assess obstacle-negotiating gait (ONG) and explore its related physical factors. Subjects comprised 571 community-dwelling persons (195 men and 376 women) age>or=65 years and without cognitive impairment. ONG, timed up and go (TUG), and stair-climbing were measured and their relations were assessed. Walking ability (usual walking speed=UWS and daily walking steps=DWS), balance deficiency, and musculoskeletal function (handgrip strength and bone health) were examined objectively to explore ONG related physical factors. Logistic regression and general linear model analysis were performed with adjustment for age and body mass index (BMI). Time of ONG was 7.72s in men and 8.93s in women. Logistic regression analysis showed that the fast tertiles of ONG corresponding to the fast levels of TUG and stair-climbing had the highest odds ratio values. General linear model analysis showed that ONG was significantly associated with UWS and balance deficiency (no) in men, and with UWS, handgrip strength, DWS and balance deficiency (no) in women. Overall, ONG was considerably associated with falls screening tools and its determinants were also the main causes of falls. Training in ONG could be an attractive strategy for the prevention of falls among the elderly., (Copyright (c) 2009 Elsevier Ireland Ltd. All rights reserved.)

Aim: The Trail Making Test (TMT) is used in Western countries as an indicator of executive function, but there is little information regarding its use in Japan. Our previous initial study showed that the TMT was significantly associated with mobility-related functions among elderly Japanese living in the community and in this study, we increased the number of participants to clarify that result., Methods: The TMT, comprised of two parts (part A and part B), was administered to 493 subjects aged 65 years or older (164 men, 329 women). Our assessment used the time difference (DeltaTMT) between parts B and A, and eight physical performance indicators: four of preventive care (usual walking speed, timed Up & Go [TUG], one-leg standing balance and handgrip strength) and four movement parameters (maximum walking speed, dual-task TUG, stair climbing and obstacle-negotiating gait)., Results: The median DeltaTMT score was 64.01 s for men and 65.56 s for women. The DeltaTMT score increased with age, and there was no difference between sexes. Multinomial logistic regression analysis showed that a poor DeltaTMT was related to low tertiles for all physical performances. It related to the intermediate tertile of TUG and maximum walking speed, dual-task TUG, stair climbing and obstacle-negotiating gait., Conclusion: The TMT reflects complex walking performance, so it can be a useful synthetic indicator for health programs promoting independence in elderly Japanese.

Aim: The purpose of this study is To examine the influence of age on muscle mass in a Japanese population for health promotion., Methods: Subjects were 4,003 community-dwelling Japanese men and women. We employed four-frequency bioelectrical impedance analysis to estimate upper and lower limbs, trunk and whole body muscle mass., Results: Men showed significantly more muscle mass in all parts of the body compared to women. There was a curvilinear relationship between age and muscle mass in both men and women. For all parts of the body, the slope of the regression line between age and muscle mass was greater in men than women. The changes in muscle mass with advancing age were different in each part of the body. In the upper limbs, there was little change with advancing age in both men and women. In the lower limbs, the decrease in muscle mass began after two decades, with the reduction in this muscle mass the greatest of all parts of the body with advancing age. In the trunk, the slope of the regression line increased from the second to the fifth decade, after which the slope decreased., Conclusions: These findings indicated that lower limb muscle mass was the first to begin to decrease and also showed the greatest decrease. For health promotion, it was seen as important to maintain muscle mass from at least middle age, with particular emphasis on the lower limbs.

Purpose: The masticatory ability of community-dwelling elderly persons is often evaluated using subjective questionnaires. However, an objective evaluation would clearly be beneficial so that problems can be adequately addressed. The purpose of this study was to determine the utility of color-changeable chewing gum to evaluate masticatory ability in community-dwelling elderly persons., Methods: We analyzed the reliability of the chewing gum for assessment of masticatory ability in 5 individuals who were examined on 5 occasions at 2 hour intervals. We also analyzed the validity of color-change of the chewing gum for assessment of masticatory ability by means of a four-part examination of 210 community-dwelling elderly persons. The four parts were as follows: evaluation of masticatory ability with the gum, determination of the number of residual teeth, measurement of maximum bite force, and a questionnaire in which participants were asked to assess their own masticatory ability. Color changes in the gum after chewing were measured with a color-reader and quantified with the a* color space defined by the Commission Internationale de l'Eclairage. A higher a* value indicates a higher degree of mastication., Results: With respect to reliability, the coefficient of variation for the a* of the color-changeable chewing gum was 2.15-3.75%. There were no significant a* value differences between men and women in any age group. Decrease with age was significantly only in women. With respect to validity, the a* value of the color-changeable chewing gum had a significant positive correlation with the number of residual teeth and maximum bite force in both men and women. In addition, men and women who reported good masticatory ability in response to the questionnaire had significantly high a* values., Conclusion: We found the use of color-changeable chewing gum to be a highly reliable and valid method for evaluating masticatory ability. Our results indicate that this method could be useful for monitoring masticatory ability in the community-dwelling elderly.

Aim: The Trail Making Test (TMT) has been studied in western countries as an indicator of executive function, but there are few data on the TMT in Japan. This study was conducted to assess the characteristics of TMT and to explore the effect of TMT on physical function among community-dwelling elderly Japanese to promote health in the aged., Methods: Subjects were 175 individuals aged>or=65 years (57 men and 118 women). The TMT test which consists of two parts (part A and part B)was performed. 8 physical function items such as indicators of functional decline (usual walking speed, timed up-and-go (TUG), one-leg standing balance, and handgrip strength), and indicators of mobility (maximum walking speed, TUG with cup, stair-climbing, obstacle-negotiating gait) were measured. For this analysis, we used a difference score defined as TMT calculated as the difference between times (part B-part A=TMT). Data were analyzed by sex respectively., Results: The median TMT value was 58.61 seconds in men and 65.67 seconds in women. TMT value increased with age and there was no difference between men and women in absolute values. multinomial logistic regression analysis showed that poor TMT was related to low tertiles of TUG, handgrip strength, MWS, TUG with cup, stair-climbing and obstacle-negotiating gait, and to intermediate tertile of MWS with adjustment for age and sex., Conclusion: TMT was significantly associated with mobility-related functions, suggesting that TMT, as the indicator of executive function, should be considered to be included in the test batteries for evaluating older people.

Aim: The purpose of this study was to determine the best way to indicate muscle mass of community-dwelling elderly men and women., Methods: We recruited 398 elderly men and women (age range, 61-96, years) who use a community center and live in a metropolitan suburb. We measured appendicular and whole body muscle mass by using bioelectrical impedance analysis and analyzed our results in relation to indicators of physical function. We assessed muscle mass with no adjustment, adjustment by body mass, and adjustment by body height., Results: In men, appendicular muscle mass adjusted by body mass correlated significantly with 10-m obstacle walking time, and whole body muscle mass adjusted by body mass correlated significantly with all indicators physical function except five chair stands and maximum walking speed. In women, appendicular and whole body muscle mass adjusted by body mass correlated significantly with all indicators of physical function. However, appendicular and whole body muscle mass adjusted by body height were unrelated to indicators of physical function., Conclusions: Only whole body muscle mass adjusted by body mass was related to physical function in both men and women. This finding suggests that whole body muscle mass adjusted by body mass is the best indicator of muscle mass in community-dwelling elderly persons.

Isoprostanes comprise a class of membrane lipid metabolites produced during oxidative stress, including asthma, chronic obstructive pulmonary disease, and cystic fibrosis. They are widely recognized to evoke a variety of biological responses in airway and pulmonary vascular smooth muscle, lymphatics, and innervation. However, their effects on airway epithelium are largely unstudied. We examined the electrophysiological responses evoked by several different isoprostane species in bovine airway epithelium using the Ussing chamber technique. The E-ring isoprostanes 15-E(1t)-IsoP and 15-E(2t)-IsoP evoked a substantial increase in short-circuit current (I(SC)), whereas four different F-ring isomers were ineffective. 15-E(2t)-IsoP-evoked I(SC) was mimicked by the prostaglandin E(2)-selective prostanoid receptor (EP)-agonist prostaglandin E(2) but not by agonists of EP(1)/EP(3)-, FP-, or TP receptors (sulprostone, fluprostenol, and U46619, respectively). This response was significantly reduced by the EP(4)-receptor blocker GW627386 but not by blockers of other prostanoid receptors (ICI 192,605 [TP-selective], SC19220 [EP(1)-selective], AH6809 [DP/EP(1)/EP(2)-selective], and AL8810 [FP-selective]). 15-E(2t)-IsoP-evoked I(SC) was reduced by blockers of Cl(-) channels (niflumic acid and 5-nitro-2-(3-phenylpropylamino)-benzoic acid), of Na(+)/K(+)/2Cl(-) co-transport (furosemide and bumetanide), of adenylate cyclase (MDL 12,330A), or of guanylate cyclase (1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one) but not by blockers of Na(+) conductances (amiloride). We conclude that 15-E(2t)-IsoP activates a transepithelial Cl(-) conductance in bovine airway epithelium through an EP(4) receptor coupled to adenylate cyclase and soluble guanylate cyclase.

Quantification of CD4+ T lymphocytes and viral RNA in patients with HIV/AIDS. Levels of progression markers (viral load and CD4+ T lymphocytes) in 410 patients with HIV/AIDS that were in different clinical stages of the disease and under different therapeutic schemes were quantified. The objective was to determine the correlation between values of progression markers and clinical stage of the patients. Commercial methodologies for the quantification of lymphocytes, subpopulations and circulating viral RNA were used. Results indicate that there was a correlation between low CD4+ values and high viral load in patients with antiretroviral treatment but not in patients without treatment. Furthermore, analysis of 1,208 samples processed during 1999 showed that 46% of the patients had less than 200 CD4+ T lymphocytes/mL blood and more than 500 copies of circulating viral RNA. Implications of these results in public health in Mexico are discussed.

Cecal and environmental samples were collected and examined for the presence of Salmonella on a farm where a high prevalence of Salmonella blockley in chickens was observed. Of 895 cecal and 525 environmental samples examined, 242 (27.0%) and 202 (38.5%) samples, respectively, yielded S. blockley. From the analysis of plasmid profile patterns, 11 different plasmid profile patterns were found among 444 isolates, with plasmid patterns c and d the most frequent among the isolates from ceca and the environment. Salmonella blockley was isolated from environmental samples such as floor litter, walls, drinking water, waste water, dust, and soil collected when barns were occupied and was positive in drinking water, waste water, and soil when samples were collected from empty barns with occupied neighboring barns, but it was negative in all environmental samples with the exception of soil when the environmental samples were collected from empty barns with empty neighboring barns.

Cecal contents of 2,345 broiler chickens consisting of 28 flocks originated from 12 farms were examined for the prevalence of Salmonella to know the actual status of infection with Salmonella in the chicken flocks. Salmonella was isolated from 336 (14.3%) samples. From these isolates, eight serovars were identified. Of the 336 Salmonella isolates, 242 (72.0%) were serotyped as S. Blockley, 60 (17.9%) S. Hadar, 15 (4.5%) S. Bredeney, nine (2.7%) S. Schwarzengrund, four (1.2%) S. Anatum, three (0.9%) S. Enteritidis, two (0.6%) S. Ohio, and one (0.3%) S. Livingstone. The same serovars of Salmonella were repeatedly found in the chickens from the same farms. S. Typhimurium and S. Enteritidis were detected in pooled broken eggshell samples collected from the hatchery. Analysis of plasmid profiles revealed 11 patterns of S. Blockley and seven patterns of S. Hadar. Strains of the same plasmid profiles of S. Blockley were isolated repeatedly from the same farm over one year after the first isolation.

Background: Oral sulindac is known to reduce polyps in patients with familial adenomatous polyposis (FAP). The authors speculated that rectal administration of indomethacin would be effective therapy for adenomas in the rectal remnant of FAP., Methods: Eight patients with FAP who had been treated by total colectomy with ileorectal anastomosis were administered an indomethacin suppository (50 mg) once or twice daily during a period of 4 or 8 weeks. The number of polyps at the same site within the rectum was counted under proctoscopy prior to, at the end of, and after the treatment. In four patients, proliferative activity of the rectal mucosa was assessed by immunohistochemical staining for MIB-1., Results: In six of the eight patients who initially had ten or more polyps, the number of polyps decreased to fewer than five, whereas such a decrease could not be observed in the remaining two patients. In the six patients, the number of polyps increased after indomethacin was discontinued. The proliferative activity of the rectal mucosa was higher at the end of treatment than it was prior to indomethacin administration., Conclusions: Indomethacin suppositories may be effective in the management of rectal adenomatosis in patients with FAP.

To determine, whether 558Thr in the carboxyl-terminal domain of moesin is phosphorylated in cells other than platelets, rabbit phosphorylation state-specific antibodies were made to the chemically phosphorylated synthetic hexapeptide KYKpTLR of the moesin sequence, as well as to the unphosphorylated form. The affinity-purified antibody populations were specific for either the phosphorylated or the unmodified peptide conjugated to BSA. Site-specific phosphorylation of moesin is detected in RAW macrophages by Western blot analysis, and immunofluorescence studies demonstrate that phosphorylated moesin is localized in filopodial protrusions. After pretreatment with the phosphatase inhibitor calyculin A, a similar effect to that seen in platelets in found, namely a substantial increase in moesin phosphorylation at 558Thr and redistribution of phospho-moesin together with F-actin into one or more ring-like structures in the cytoplasm, presumably due to binding of phosphorylated moesin to F-actin.

In the chicken, enkephalin-immunoreactive cells and nerve fibers are distributed in the ultimobranchial glands, which consist of C-cell groups and cyst structures. Ultrastructural features of the enkephalin cells and nerve fibers were examined by immuno-electron microscopy using both the streptavidin-biotin-peroxidase method and the protein A-colloidal gold method. Immunoreactivity for enkephalin was located on the secretory granules of C cells. In 1-day-old chickens, three types of C cells were distinguished on the basis of their granule size. Type-I cells were filled with large secretory granules (200-600 nm in diameter). These elements represented a majority of the C-cell population. Type-II cells contained medium-sized granules (100-280 nm in diameter). Type-III cells displayed small secretory granules (60-200 nm in diameter). The latter cells were elongate or irregular in shape and frequently extended cytoplasmic processes into the connective tissue stroma or contacted other C cells. Enkephalin-immunoactivity was revealed by dense deposits of immunogold particles on the secretory granules of type-II and type-III cells. There were only a few type-I cells showing immunoreactivity for enkephalin. A double immunogold labeling procedure demonstrated that calcitonin and enkephalin were colocalized in the same secretory granules of type-I and type-II cells. Type-III cells were devoid of immunoreactivity for calcitonin. Enkephalin-immunoreactive nerve fibers were characterized by the presence of granular vesicles, 60-160 nm in diameter, and frequently established direct contact with the surface of C cells.(ABSTRACT TRUNCATED AT 250 WORDS)

The immunoreactivity of a monoclonal antibody against cell suspensions from guinea pig adrenal glands was examined at light- and electron-microscopic levels. In addition to the cell surface membrane of adrenocortical cells, the antibody labeled specific sites in the pancreas, liver and testis, but did not label any of the other tissues examined. In the pancreas, microvilli-like processes and the cell surface membrane of centroacinar cells were immunoreactive to the antibody. The microvilli of interlobular duct cells and pancreatic duct cells were also immunoreactive. In the liver, bile canalicular microvilli of hepatocytes were exclusively labeled. Membrane structures of cell organelles, mainly mitochondria, in testicular Leydig cells were also labeled. Immunoblot analysis showed that the monoclonal antibody bound to two common bands at molecular weights of approximately 62 kDa and 110 kDa in the pancreas, liver, testis, and adrenal gland. The two bands reacted with the digoxigenin-conjugated lectin, Sambucus nigra agglutinin (SNA), which recognizes sialic acid linked alpha (2-6) to galactose. Reaction patterns of SNA in the pancreas, liver and testis were similar to those of the monoclonal antibody; pancreatic centroacinar cells and interlobular duct cells, hepatocyte bile canaliculi and testicular Leydig cells were densely stained with SNA. Thus, the monoclonal antibody recognizes two common membrane glycoproteins containing sialic acids in the pancreas, liver, testis and adrenal cortex.

Immunogenicity, eliciting antigenicity of MT-141 and its cross-reactivity with other beta-lactam antibiotics were studied in mice, guinea pigs and rabbits. The results were as follows. Injections of MT-141 failed to produce IgE-type antibody in mice but injections of the MT-141 conjugated to rabbit serum albumin produced a trace of IgE-type antibody. No antibody was produced in the guinea pigs immunized with the MT-141 conjugated to rabbit serum albumin in alum or Freund's complete adjuvant. The conjugated MT-141 also failed to elicit anaphylactic shock in the immunized guinea pigs. The subcutaneous treatments with MT-141 in Freund's complete adjuvant produced an amount of hemagglutination antibody in rabbits. The intravenous treatments with MT-141 produced no antibody in rabbits. When rabbits were subcutaneously immunized with the MT-141 conjugated to rabbit serum albumin in Freund's complete adjuvant, production of specific antibody in the rabbits was demonstrated by observations of passive cutaneous anaphylaxis and hemagglutination. The results of hapten-induced inhibition of passive hemagglutination, passive cutaneous anaphylaxis, anaphylactic shock and hemagglutination by using conjugates of antibiotics and rabbit serum albumin as immunogens and conjugates of antibiotics and bovine gamma-globulin as eliciting antigen showed that MT-141 did not cross-react with other antibiotics. MT-141 did not cause the in vitro direct Coombs' reaction in the human blood even at a high concentration of 160 mg/ml. It is concluded from these results that immunological activity of MT-141 preparation is weak.

In ACD solution which contains PEP and sucrose was used as a preservative solution as well as a medium for rejuvenating depleted erythrocytes. 2,3-DPG and ATP in whole blood or red cell concentrates were increased effectively by incubating the cells with the solution at 37 degrees C for 30-60 min. The transport of PEP through the erythrocyte membrane was essential to the increase of 2,3-DPG and ATP. During storage of the cells at 4 degrees C in the presence of PEP, no increase of 2,3-DPG and ATP was observed because no transport of PEP into the cells occurred. By incubation at 37 degrees C for 30 min at the end of various storage periods, however, the levels of ATP and 2,3-DPG in the cells were raised. Addition of ascorbic acid, purine or purine nucleosides to the ACD-sucrose-PEP solution improved the PEP effect of maintaining ATP and/or 2,3-DPG in erythrocytes during storage at 4 degrees C.

A fertility study of MT-141 was performed in SD rats with the intramuscular (i.m.) injections at the dose levels of 400, 800 and 1,600 mg/kg/day. The male rats were injected with MT-141 for 63 days before mating and during the mating period, while the female rats were injected with MT-141 from the 14th day before mating up to the day 7 of gestation. All pregnant rats were sacrificed on day 20 of gestation followed by external, visceral and skeletal observations of their fetuses. The results are summarized as follows. The suppression of body weight gain was observed in males given above 800 mg/kg/day i.m. and in females of all treated groups during early period of gestation. However, no significant differences were found between treated groups and the control with regard to copulation rate and conception rate. Though no defects were observed for visceral and skeletal specimens in the fetuses of treated groups, MT-141 produced a delayed ossification of forelimbs in the fetuses at the doses above 800 mg/kg/day and of sternebrae at the dose of 1,600 mg/kg/day. It is concluded from the above-mentioned results that the maximal "no 'effective" dose of MT-141 on the fertility is above 1,600 mg/kg/day i.m. in parental rats and less than 800 mg/kg/day i.m. for the fetuses.