Katsumi Murakami

Cellular localization of inflammatory cytokines in human glomerulonephritis.

We evaluated the expression of inflammatory cytokines in renal tissues obtained from 45 patients with several types of glomerulonephritis. Immunofluorescence studies with specific antibodies to interleukin (IL)-1α, IL-1β, IL-6, tumour necrosis factor (TNF)-α, and TNF-β showed intense cytoplasmic staining in the glomeruli and interstitium. Cells positive for these cytokines were found frequently in tissue from patients with lupus nephritis (WHO Class IV) and membranoproliferative glomerulonephritis, and, to a lesser extent, in tissue from patients with mesangial proliferative glomerulonephritis, Henoch-Schönlein purpura nephritis, and minimal change nephrotic syndrome. Most of these cells were dual-stained with a monoclonal antibody to monocytes-macrophages. In situ hybridization for cytokine mRNA, combined with immunoperoxidase staining for monocytes-macrophages, detected IL-1α, IL-6, and TNF-α mRNA in monocytes-macrophages infiltrating the glomeruli and interstitium. Occasionally, there was weak or moderate immunostaining for IL-1α, IL-6, and TNF-α in the glomerular mesangial and epithelial cells, but in situ hybridization signals were rarely found in these loci. These findings suggest that infiltrating monocytes-macrophages, rather than resident glomerular cells, are the major source of inflammatory cytokines in human glomerulonephritis.

Urinary transforming growth factor-β in patients with glomerular diseases

Abstract. We measured the urinary levels of active transforming growth factor-β (TGF-β) by enzyme-linked immunosorbent assay in 12 healthy controls and 42 patients with various glomerular diseases, including mesangial proliferative (IgA nephritis, Henoch-Schönlein purpura nephritis, and IgA-negative mesangial proliferative glomerulonephritis) and non-proliferative (minimal change nephrotic syndrome and focal glomerulosclerosis) types. Urinary TGF-β, expressed as a ratio to urinary creatinine (ng/mg creatinine), was elevated in patients with IgA nephritis and focal glomerulosclerosis, and was significantly higher than in patients with other types of glomerular diseases and healthy controls. There was a significant correlation between urinary TGF-β levels and the grade of interstitial fibrosis. Among patients with proliferative-type disease, urinary TGF-β was significantly correlated with the grade of mesangial matrix increase and the magnitude of proteinuria. The relationship between urinary TGF-β levels and the immunostaining intensity of TGF-β in the glomeruli was not significant. These results indicated that urinary TGF-β reflects the grade of interstitial fibrosis in glomerular diseases and also the mesangial matrix increase in proliferative-type glomerulonephritis. Measuring TGF-β levels in the urine might be helpful in monitoring patients with some types of glomerular disease.

Immunohistochemical localization of C3d fragment of complement and S-protein (vitronectin) in normal and diseased human kidneys: association with the C5b-9 complex and vitronectin receptor

The localization of C3d, a fragment produced by C3 activation and S-protein (vitronectin), a regulatory factor of C5b-9, was studied immunohistochemically in normal human kidney and renal biopsies from patients with several types of glomerulonephritis. Immunofluorescent staining of the normal kidneys showed that C3d was present along the glomerular basement membrane (GBM), tubular basement membrane (TBM) and arterioles, and that S-protein was present in the GBM, mesangium, TBM, and arterioles. Immunoelectron microscopy of isolated basement membranes showed that C3d was localized exclusively on the epithelial side of the GBM, and that S-protein was present along both the epithelial and endothelial sides. In nephritic tissues, glomerular staining of C3d, C5b-9, and S-protein was increased when compared with that in normal tissues. S-protein, frequently co-localized with C3d and C5b-9 neoantigen, was intensely positive in the immune deposits of glomerular capillaries and the mesangial area, overlapping the background staining of GBM and mesangial matrix. S-protein and its receptor were occasionally co-localized in the glomeruli. These findings indicate that C3d and S-protein are normally present in the glomeruli. Co-staining of C3d, C5b-9 neoantigen, and S-protein within the immune deposits of nephritic kidneys suggests in situ binding of S-protein to locallyformed C5b-9 complex, or merely co-distribution of S-protein with the complex, rather than trapping of large molecular SC5b-9 complex from the circulation.

High-pitched breath sounds indicate airflow limitation in asymptomatic asthmatic children

Background and objective:  Asthmatic children may have airway dysfunction even when asymptomatic, indicating that their long‐term treatment is less than optimal. Although airway dysfunction can be identified on lung function testing, performing these tests can be difficult in infants. We studied whether breath sounds reflect subtle airway dysfunction in asthmatic children.

The effect of Lactobacillus brevis KB290 against irritable bowel syndrome: a placebo-controlled double-blind crossover trial

BackgroundIrritable bowel syndrome (IBS) is a functional disorder of the digestive tract that causes chronic abdominal symptoms. We evaluated the effects of Lactobacillus brevis KB290 (KB290), which has been demonstrated to be effective at improving bowel movements and the composition of intestinal microflora, on IBS symptoms.MethodsWe performed a placebo control double-blind cross matched trial. Thirty-five males and females (aged 6 years and above) who had been diagnosed with IBS according to the Rome III criteria were divided into 2 groups, and after a 4-week pre-trial observation period, they were administered test capsules containing KB290 or placebo for 4 weeks (consumption period I). Then, the capsule administration was suspended for 4 weeks in both groups (washout period), before the opposite capsules were administered for a further 4 weeks (consumption period II). Fecal samples were collected on the first day of the pre-consumption observation period, the last day of consumption period I, the last day of the washout period, and the last day of consumption period II. In addition, the subjects’ IBS symptoms and quality of life (QOL) and any adverse events that they experienced were evaluated.ResultsNo significant difference in IBS symptoms was noted among the various periods. However, the mean QOL scores were improved during the test capsule consumption.The frequencies of watery and mushy feces were significantly lower in the test capsule consumption period than during the pre-consumption observation period, and the frequency of abdominal pain was significantly reduced in the test capsule consumption period compared with the other periods.The frequency of the genus Bifidobacterium was significantly higher, and that of the genus Clostridium was significantly lower, after the test capsule consumption than after the placebo consumption. The frequencies of the genera Lactobacillus, Bacteroides, and Enterococcus were also investigated, but no differences in their frequencies were detected between the placebo and test capsule consumption periods.ConclusionsProbiotics, the safety of which has been established, are used widely in various foods and can now be purchased readily. The results of the present study suggest that KB290 is useful for early intervention in IBS.

Changes in the highest frequency of breath sounds without wheezing during methacholine inhalation challenge in children.

A breath sound analyser was used to detect bronchoconstriction without wheezing during methacholine inhalation challenge in children. The highest frequency of inspiratory breath sounds increased significantly during bronchoconstriction and decreased after inhalation of a bronchodilator. The highest frequency of inspiratory breaths sounds was correlated with bronchial reactivity.

Proto-oncogene expression in human glomerular diseases

The expression of the protein products and mRNA of c‐fos, c‐myc, p53, and c‐raf was examined in normal renal tissues and biopsy specimens from 73 patients with various glomerular diseases. Immunofluorescent staining showed that there were cell nuclei stained for c‐Fos, c‐Myc, and p53, and cytoplasm positive for c‐Raf, in the glomeruli of patients with proliferative types of glomerulonephritis, including IgA nephritis and lupus nephritis, and in patients with focal glomerular sclerosis. Glomerular expression of c‐fos and c‐myc mRNA was detected by in situ hybridization. The number of proto‐oncogene‐positive glomerular cells was significantly higher in lupus nephritis, IgA nephritis, and focal segmental sclerosis, as compared with minimal change nephrotic syndrome and normal specimens. In IgA nephritis, the population of glomerular cells positive for c‐Fos and c‐Myc and the grade of c‐Raf immunoreactivity were significantly correlated with the proportion of proliferating cell nuclear antigen (PCNA)‐positive glomerular cells, with histological grading of mesangial hypercellularity and matrix increase, and with the magnitude of proteinuria. These data indicate that proto‐oncogene expression is associated with mesangial proliferation and matrix expansion in proliferative types of glomerulonephritis and in focal glomerular sclerosis.

A New Modality Using Breath Sound Analysis to Evaluate the Control Level of Asthma

BACKGROUND Reliable symptom assessment is essential in asthma management. We developed new technology for analyzing breath sounds and assessed its clinical usefulness for monitoring asthmatic children. METHODS Eighty asthmatic children and 59 non-asthmatic children underwent breath sound analysis in an asymptomatic state. Their asthma control was assessed by the Asthma Control TestTM or Childhood ACTTM scores and divided into two groups, namely, well-controlled (perfect) (n = 19) and not well-controlled (not perfect) (n = 61). Breath sounds were recorded using two sensors, located on the right anterior chest and trachea. We calculated the acoustic transfer characteristics between the two points, which indicated the relationship between frequencies and attenuation during breath sound propagation. Two indices of sound parameters, the chest wall sound index (CWI) and the tracheal sound index (TRI), were calculated from the transfer characteristics and tracheal sounds. We also developed a new parameter, the breath sound index (BSI), on a 2-dimensional diagram of CWI and TRI and tried to determine whether BSI may clarify asthma control better than CWI or TRI alone. RESULTS There was a significant difference in TRI and BSI between asthmatic and non-asthmatic children (p = 0.007, p < 0.001). There was a significant difference in CWI and TRI between the well-controlled and not-wellcontrolled groups (p < 0.001). BSI discriminated between the two groups accurately (p < 0.001). The sensitivity and specificity of BSI for asthma control were 83.6% and 84.2%, respectively. CONCLUSIONS Asthma control could be evaluated using a new index calculated from breath sound analysis.BACKGROUND Reliable symptom assessment is essential in asthma management. We developed new technology for analyzing breath sounds and assessed its clinical usefulness for monitoring asthmatic children. METHODS Eighty asthmatic children and 59 non-asthmatic children underwent breath sound analysis in an asymptomatic state. Their asthma control was assessed by the Asthma Control TestTM or Childhood ACTTM scores and divided into two groups, namely, well-controlled (perfect) (n = 19) and not well-controlled (not perfect) (n = 61). Breath sounds were recorded using two sensors, located on the right anterior chest and trachea. We calculated the acoustic transfer characteristics between the two points, which indicated the relationship between frequencies and attenuation during breath sound propagation. Two indices of sound parameters, the chest wall sound index (CWI) and the tracheal sound index (TRI), were calculated from the transfer characteristics and tracheal sounds. We also developed a new parameter, the breath sound index (BSI), on a 2-dimensional diagram of CWI and TRI and tried to determine whether BSI may clarify asthma control better than CWI or TRI alone. RESULTS There was a significant difference in TRI and BSI between asthmatic and non-asthmatic children (p = 0.007, p < 0.001). There was a significant difference in CWI and TRI between the well-controlled and not-well-controlled groups (p < 0.001). BSI discriminated between the two groups accurately (p < 0.001). The sensitivity and specificity of BSI for asthma control were 83.6% and 84.2%, respectively. CONCLUSIONS Asthma control could be evaluated using a new index calculated from breath sound analysis.

Evaluation of airway responsiveness using colored three-dimensional analyses of a new forced oscillation technique in controlled asthmatic and nonasthmatic children

BACKGROUND Bronchodilator response (BDR) is routinely used in asthma management. A new forced oscillation technique (FOT) is able to quickly measure respiratory system resistance (Rrs) and reactance (Xrs) at each tidal breath phase. The present study evaluated bronchial changes by using the new FOT. METHODS Respiratory resistance and reactance were measured using FOT in 132 children (age, 10.86±4.78 years; M:F=88:44), including asthmatic (n=98) and nonasthmatic children (n=34), pre- and post-bronchodilator inhalation in an asymptomatic state. Whole-breath or within-breath changes in Rrs and Xrs were measured and compared pre- and post-bronchodilator inhalation and between each group. All patients performed spirometry and forced expiratory nitric oxide pre- and post-bronchodilator inhalation. RESULTS Spirometric parameters showed significant positive changes at V50 and V25 in both groups; however, these changes were not significantly different between the groups. eNO was significantly higher in the asthmatic group than in the nonasthmatic group; however, there was no significant change pre- and post-inhalation in either group. Rrs in the asthma group was significantly higher in the expiratory phase than in the inspiratory phase. Rrs and Xrs before and after bronchodilator inhalation were significantly different in the asthma group alone, except for the expiratory-inspiratory phase of each of these parameters. Changes in Rrs and Xrs at 5Hz (R5 and X5) in a whole-breath and the inspiratory phase were significantly different between the groups. CONCLUSIONS Changes in X5 and R5 reflect bronchial reversibility. The new FOT is useful for asthmatic children.

Japanese clinical guidelines for chronic pain in children and adolescents.

Chronic pain is a common problem in pediatric practice. The prevalence of chronic pain in children is >30%. Because pain indicates emotional expression as well as the physiological reaction toward infection, injury, and inflammation, both physiological and psychological assessments are essential to determine primary interventions for chronic pain. The Japanese Society of Psychosomatic Pediatrics Task Force of clinical practice guidelines for chronic pain in children and adolescents compiled clinical evidence and opinions of specialists associated with the primary care of pediatric chronic pain in the Japanese ‘clinical guidelines for chronic pain in children and adolescents’ in 2009, which are presented herein. The guidelines consist of three domains: general introduction to chronic pain; chronic abdominal pain; and chronic headache. Each section contains information on the physiological mechanism, psychological aspects, assessment methods, and primary interventions for pediatric chronic pain. These guidelines are expected to help disseminate knowledge on primary interventions for chronic pain in children and adolescents.