Masaharu Shinkai

Clarithromycin Delays Progression of Bronchial Epithelial Cells from G1 Phase to S Phase and Delays Cell Growth via Extracellular Signal-Regulated Protein Kinase Suppression

ABSTRACT The nonsteroidal anti-inflammatory drugs have been shown to support cytoprotection of cells by shifting cells toward a quiescent state (G0/G1). Extracellular signal-regulated kinase (ERK) is required for cells to pass from G1 phase into S phase, and macrolide antibiotics can inhibit ERK1/2 phosphorylation. However, previous reports suggest that macrolide antibiotics do not affect cell growth in bronchial epithelial cells. Therefore, we studied normal human bronchial epithelial (NHBE) cells to determine whether clarithromycin (CAM) suppresses ERK, delays bronchial epithelial cells from progressing to S phase, and delays cell growth. Exposure to CAM at 10 μg/ml daily over 4 days irreversibly decreased the cell proliferation with and without growth supplements (P < 0.0001). CAM also inhibited ERK1/2 phosphorylation over the first 90 min of exposure (P < 0.05 for 30 min, P < 0.0001 for 60 min, and P < 0.01 for 90 min) and decreased the ratio of phosphorylated ERK1/2 (pERK1/2) to total ERK1/2 (tERK1/2) (P < 0.0001). Incubation with CAM for 48 h increased the proportion of cells in G1 phase (means ± standard deviations) from 63.5% ± 0.9% to 79.1% ± 1.4% (P < 0.0001), decreased that in S phase from 19.8% ± 1.2% to 10.0% ± 2.1% (P < 0.01), and decreased that in G2/M phase from 16.7% ± 0.4% to 11.0% ± 0.8% (P < 0.001). In contrast, the ratio of pMEK1/2 to tMEK1/2 was not altered after exposure to CAM. These results suggest that macrolide antibiotics can delay the progression of NHBE cells from G1 phase to S phase and can slow cell growth, probably through the suppression of ERK1/2.

Clarithromycin therapy for patients with Cystic Fibrosis: A randomized controlled trial

The clinically significant actions of oral azithromycin in modifying progressive cystic fibrosis (CF) lung disease have been well documented. In vitro and clinical data suggests that clarithromycin has immunomodulatory properties similar to other 14‐member macrolides, however two previously reported short term, open label trials of clairthromycin in small numbers of patients with CF failed to show significant benefits in modifying lung function or inflammation. We performed an international double blind, cross‐over trial in which 63 subjects with CF were studied while receiving either placeo or 500 mg oral clarithromycin twice daily for 5 months, with a 1‐month wash‐out. The primary efficacy end point was the change in lung function (FEV1 and FVC) during the clarithromycin treatment period compared to placebo treatment. Secondary efficacy end points included; quality of life, number of pulmonary exacerbations, height and weight, sputum inflammatory mediator content, sputum transportability and surface properties, bacterial flora, nasal potential difference, and breath condensate. No significant difference in either the primary efficacy end point or any secondary end point was seen during the period of clarithromycin treatment compared to those seen during placebo administration. We conclude that clarithromycin is not effective in treating CF lung disease. Pediatr Pulmonol. 2012; 47:551–557.

Immunomodulatory Effects of Macrolide Antibiotics

Macrolide antibiotics have been used for over 50 years to treat bacterial infections. Over the past 20 years, macrolide antibiotics have also been used to treat some chronic inflammatory airway diseases by virtue of their immunomodulatory properties. Macrolides have been reported to down-regulate prolonged inflammation or hyperinflammation by effects on cellular immunity, suppressing the production of pro-inflammatory cytokines and reactive oxygen species, blocking the activation of nuclear transcription factors, inhibiting neutrophil activation and mobilization, accelerating neutrophil apoptosis, and improving mucus clearance. Long-term, low-dose macrolide therapy has dramatically improved survival and decreased the frequency of exacerbation for persons with diffuse panbronchiolitis (DPB). In studies using azithromycin or clarithromycin, persons with cystic fibrosis (CF) had significantly improved lung function and fewer respiratory exacerbations compared with those taking a placebo. Macrolides are now first line therapy for DPB and recommended for the therapy for CF. Recent guidelines by the Japanese Ministry of Health and Welfare recommend for adults, erythromycin (EM) 400 or 600 mg/d, clarithromycin (CAM) 200 or 400 mg/d, roxithromycin (RXM) 150 or 300 mg/d, or azithromycin (AZM) 250 or 500 mg/d be given for at least 6 months and for up to 2 years. A clinical response is usually seen by the third month of therapy, and side effects are few and generally self-limited.

HO-1 inhibits IL-13-induced goblet cell hyperplasia associated with CLCA1 suppression in normal human bronchial epithelial cells.

Mucus hypersecretion and goblet cell hyperplasia are common features that characterize asthma. IL-13 increases mucin (MUC) 5AC, the major component of airway mucus, in airway epithelial cells. According to the literature, IL-13 receptor activation leads to STAT6 activation and consequent induction of chloride channel accessory 1 (CLCA1) gene expression, associated with the induction of MUC5AC. Heme oxygenase-1 (HO-1) is an enzyme that catalyzes oxidation of heme to biliverdin, and has anti-inflammatory and anti-oxidant properties. We examined the effects of HO-1 on mucin production and goblet cell hyperplasia induced by IL-13. Moreover, we assessed the cell signaling intermediates that appear to be responsible for mucin production. Normal human bronchial epithelial (NHBE) cells were grown at air liquid interface (ALI) in the presence or absence of IL-13 and hemin, a HO-1 inducer, for 14 days. Protein concentration was analyzed using ELISA, and mRNA expression was examined by real-time PCR. Histochemical analysis was performed using HE staining, andWestern blotting was performed to evaluate signaling transduction pathway. Hemin (4 μM) significantly increased HO-1 protein expression (p b 0.01) and HO-1 mRNA expression (p b 0.001). IL-13 significantly increased goblet cells, MUC5AC protein secretion (p b 0.01) and MUC5AC mRNA (p b 0.001), and these were decreased by hemin by way of HO-1. Tin protoporphyrin (SnPP)-IX, a HO-1 inhibitor, blocked the effect of hemin restoring MUC5AC protein secretion (p b 0.05) and goblet cell hyperplasia. Hemin decreased the expression of CLCA1 mRNA (p b 0.05) and it was reversed by SnPP-IX, but could not suppress IL-13-induced phosphorylation of STAT6 or SAM pointed domain-containing ETS transcription factor (SPDEF) and Forkhead box A2 (FOXA2) mRNA expression. In summary, HO-1 overexpression suppressed IL-13-induced goblet cell hyperplasia and MUC5AC production, and involvement of CLCA1 in the mechanism was suggested.

Severe or life-threatening asthma exacerbation: patient heterogeneity identified by cluster analysis

Severe or life‐threatening asthma exacerbation is one of the worst outcomes of asthma because of the risk of death. To date, few studies have explored the potential heterogeneity of this condition.

Topotecan for Relapsed Small-cell Lung Cancer: Systematic Review and Meta-Analysis of 1347 Patients

Topotecan is the most reliable chemotherapy regimen for relapsed small-cell lung carcinoma (SCLC). The efficacy and adverse effects of topotecan as reported by previous studies varied greatly. The inclusion criterion was a prospective study that was able to provide data for 6-month over-all survival (OS) rate, 1-year OS rate, objective responses, and/or adverse effects of single agent topotecan as a second line chemotherapy for SCLC, written in English language as a full article. Any topotecan regimen were allowed. Binary data were meta-analyzed with the random-model generic inverse variance method. We included 14 articles consisted of 1347 patients. Pooled values were estimated as follows. Six-month OS rate: 37% (95% CI: 28–46%). One-year OS rate: 9% (95% CI: 5–13%). Response rate: 5% (95% CI: 1–8%). Six-month OS rate: 57% (95% CI: 50–64%). One-year OS rate: 27% (95% CI: 22–32%). Response rate: 17% (95% CI: 11–23%). Grade III/IV neutropenia 69% (95% CI: 58–80%). Grade III/IV thrombopenia 41% (95% CI: 34–48%). Grade III/IV anemia 24% (95% CI: 17–30%). Non-hematorogical events were rare. Chemotherapy-related death 2% (95% CI: 1–3%). In conclusion, Topotecan provided a possibly promising outcome for sensitive-relapse SCLC and poor outcome for refractory relapse SCLC. Adverse events were mainly hematological.

Depression in Japanese Patients With Chronic Obstructive Pulmonary Disease: A Cross-Sectional Study

BACKGROUND: Some investigations have revealed an association between depression and physical measurements of COPD patients in North America and Europe, but few related studies have been performed in Asia. METHODS: In this cross-sectional study, 84 consecutive, stable out-patients with COPD (mean ± SD age 72.0 ± 9.0 y, percent-of-predicted FEV1 46 ± 15%, 15 [17.9%] female) in a Japanese community hospital were recruited. “Probable depression” was defined as a score of ≥ 6 on the short-form Geriatric Depression Scale (SF-GDS). Relationships among commonly used physical measurements, SF-GDS raw score, and probable depression were evaluated with the Spearman rank correlation test, multiple linear regression analysis, logistic regression analysis, and receiver operating characteristic curves. RESULTS: Thirty-two subjects (38.1%) had probable depression. Body mass index, obstruction, dyspnea, exercise capacity index, percent-of-predicted FEV1, Modified Medical Research Council dyspnea score, 6-min walk distance, and SpO2 had: simple correlations (r 0.42–0.60, P < .001 for all) with the SF-GDS raw score; partial correlations (r 0.25–0.51, P < .05 for all) with the SF-GDS raw score after adjusting for demographic and social factors; association with probable depression in the logistic regression analysis after adjusting for demographic and social factors (P < .05 for all); and areas under the receiver operating characteristic curve of 0.72−0.84 (P < .001 for any) for probable depression. CONCLUSIONS: Physical parameters were associated with depression in our Japanese COPD out-patients.

Evaluation of the chronic obstructive pulmonary disease assessment test in Japanese outpatients

The Chronic Obstructive Pulmonary Disease (COPD) Assessment Test (CAT) is a quality‐of‐life (QOL) questionnaire that proved to correlate with St. Georges Respiratory Questionnaire. Correlations between CAT scores and other COPD parameters have not been thoroughly evaluated in Japanese outpatients.

Amrubicin for relapsed small-cell lung cancer: a systematic review and meta-analysis of 803 patients

Currently, amrubicin is permitted for relapsed small-cell lung carcinoma (SCLC) only in Japan. The efficacy and adverse effects of amrubicin as reported by previous studies varied greatly. The inclusion criterion was a prospective study that was able to provide data for efficacy and safety by the AMR single agent regimen as second-line chemotherapy for a patient with SCLC. Binary data were meta-analyzed with the random-model generic inverse variance method. We included nine articles consisted of 803 patients. The pooled three-, six-, and nine-month progression-free survival were 63% (95% CI 57–69%, I2 = 53%), 28% (95% CI 21–35%, I2 = 71%), and 10% (95% CI 6–14%, I2 = 41%), respectively. The pooled six-, 12-, and 18-month overall survival were 69% (95% CI 61–78%, I2 = 83%), 36% (95% CI 28–44%, I2 = 80%), and 15% (95% CI 8–21%, I2 = 81%), respectively. Amrubicin seemed much more beneficial for Japanese patients. However, compared to the efficacy of topotecan presented in a previous meta-analysis, amrubicin may be a better treatment option than topotecan for both Japanese and Euro-American. Adverse effects by amrubicin were almost exclusively observed to be hematological. Notably, grade III/IV neutropenia incidence was 70% and febrile neutropenia incidence was 12%.

Measurement of eNO with portable analyser might improve the management of persistent cough at primary care practice in Japan

There are some controversial reports that investigated the usefulness of exhaled nitric oxide (eNO) to predict the efficacy of inhaled corticosteroids (ICS) in chronic cough patients. Therefore, we retrospectively analysed the usefulness of eNO measurement with portable analyser to predict the requirement of ICS therapy in persistent cough (defined as lasting for 3 weeks or more) patients in Japan and investigated whether it might improve the management of persistent cough at primary care practice.