Non-Destructive Monitoring via Electrochemical NADH Detection in Murine Cells.

Nicotinamide adenine dinucleotide (NADH) is an important cofactor involved in metabolic redox reactions in living cells. The detection of NADH in living animal cells is a challenge. We developed a one-step monitoring method for NADH via an electrocatalytic reaction that uses a surface-modified, screen-printed electrode (SPE) having a redox active monolayer 4'-mercapto-N-phenlyquinone diamine (NPQD) formed by a self-assembled monolayer (SAM) of an aromatic thiol, 4-aminothiophenol (4-ATP). This electrode has a limit of detection (LOD) of 0.49 µM and a sensitivity of 0.0076 ± 0.0006 µM/µA in cell culture media, which indicates that it retains its selectivity. The applicability of this NADH sensor was demonstrated for the first time by cell viability monitoring via NADH-sensing in cell culture supernatants.

Establishment of patient-derived xenografts from patients with gastrointestinal stromal tumors: analysis of clinicopathological characteristics related to engraftment success.

Patient-derived xenografts (PDXs) can represent the heterogeneity and histological characteristics of tumors and are thus useful for testing the efficacy of anti-cancer drugs; however, PDXs are difficult to generate, especially for gastrointestinal stromal tumor (GIST). We analyzed the clinicopathologic factors associated with the successful establishment of GIST PDX in NOD.Cg-Prkdcscid IL2rgtm1Wjl/SzJ mice. We used 185 GIST tumor fragments from patients who underwent surgical resection prior to (n = 66; 35.7%) and after treatment (n = 119; 64.3%) with tyrosine kinase inhibitors. The overall success rate of PDX establishment was 17%; in univariate analysis, engraftment success was associated with after TKI treatment, larger tumor size, higher mitotic count, higher Ki-67 index, higher cellularity, presence of tumor necrosis, primary mutations in KIT exon 11, and originating from metastatic lesions. In multivariate analysis, higher Ki-67 index, after TKI treatment, and larger tumor size were independent factors for engraftment success. Immunohistochemistry in representative samples further corroborated the above results. These results will be useful in the establishment of PDX models from GISTs.

Treatment of peri-invagination lesion and vitality preservation in an immature type III dens invaginatus: a case report.

BACKGROUND: To report a case of type III dens invaginatus associated with peri-invagination periodontitis in an immature permanent mandibular central incisor with open apex, in which only the invagination area was treated and vitality was preserved. CASE PRESENTATION: A 9-year-old boy was referred complaining of pain in the mandibular left central incisor. After radiographic examination, an invagination into the pulp chamber of the tooth associated with periapical radiolucency was detected. Endodontic access was performed and the orifice was identified under a dental operating microscope. The invagination area was chemo-mechanically cleaned. After 1 week, the invagination was obturated with mineral trioxide aggregate. During the 2-year follow up period, the tooth was asymptomatic. Radiographic examination revealed significant progression of periapical healing and root development in the main root canal of the tooth. CONCLUSION: Non-surgical root canal treatment of the invagination may preserve pulp vitality, and continuous root development of the tooth.

Clinical significance of MET gene amplification in metastatic or locally advanced gastric cancer treated with first-line fluoropyrimidine and platinum combination chemotherapy.

OBJECTIVE: To investigate the clinical significance of MET gene amplification in patients with gastric cancer in the palliative setting. METHODS: MET amplification was assessed using fluorescence in situ hybridization (FISH) in 50 patients and quantitative polymerase chain reaction (qPCR) in 326 patients; 259 patients treated with first-line fluoropyrimidine and platinum were included for survival analysis. RESULTS: The results of FISH and qPCR indicated that the c-MET/CEP7 ratio was correlated with gene copy number. The optimal cutoff value for the copy number using qPCR to detect MET gene amplification with FISH was 5 (κ=0.778, P<0.001). Twenty-one out of 326 patients (6.4%) were identified asMET amplification with a copy number of >5 detected by qPCR. MET-amplified gastric cancer was associated with an Eastern Cooperative Oncology Group (ECOG) performance status (PS) score of ≥2 (33.3% vs. 10.5% P=0.007), peritoneal metastasis (76.2% vs. 46.2%, P=0.008), and elevated bilirubin levels (28.6% vs. 7.3%, P=0.006). The median overall survival (OS) and progression-free survival (PFS) were 11.9 and 5.6 months, respectively. MET-amplified gastric cancer was not associated with survival outcomes [hazard ratio (HR)=0.68, 95% confidence interval (95% CI): 0.35-1.32, P=0.254 for PFS; HR=0.68, 95% CI: 0.35-1.32, P=0.251 for OS]. CONCLUSIONS: qPCR can be used to detect MET gene amplification. MET amplification was not a predictor of poor prognosis in patients with metastatic or unresectable gastric cancer.

In Vitro Comparison of Biocompatibility of Calcium Silicate-Based Root Canal Sealers.

The aim of this study was to assess the effect of three calcium silicate-based sealers (EndoSeal MTA, Nano-ceramic Sealer, and Wellroot ST) and two epoxy resin-based sealers (AH-Plus, AD Seal) on various aspects, such as cell viability, inflammatory response, and osteogenic potential, of human periodontal ligament stem cells (hPDLSCs). AH-Plus showed the lowest cell viability on hPDLSCs in all time periods in fresh media. In set media, hPDLSCs showed no significant differences in cell viability among all the tested materials. Wellroot ST showed the highest level of cell adhesion and the morphology of attached cells. AH-plus presented a significantly higher expression of IL-6 and IL-8 than the other sealers. AD Seal and three calcium silicate sealers showed high expression of the mesenchymal stem cell markers. ALP mRNA expression showed a significant increase in time-dependent manner on all of three calcium silicate-based sealers, which do not seem to interfere with the differentiation of hPDLSCs into osteoblasts. Based on the results from this study, calcium silicate-based sealers appear to be more biocompatible and less cytotoxic than epoxy resin-based sealers. Meanwhile, further and long-term clinical follow-up studies are required.

Efficacy and retrievability of root canal filling using calcium silicate-based and epoxy resin-based root canal sealers with matched obturation techniques.

This study aimed to compare the efficiency of root canal filling procedures and the retrievability of the filling material with various sealers. Forty-three patients assigned to endodontic treatment with (i) continuous wave of condensation technique (CTW) with AH-plus (ii) single-cone technique (SCT) with EndoSeal MTA. The spent time, voids entrapping and postoperative symptoms were evaluated. To evaluate the retrievability, mandibular premolar (n = 60) were divided into four groups: AH-plus/CTW, EndoSeal MTA/SCT, MTA Fillapex/SCT and EndoSequence BC Sealer/SCT. The time required removing the filled materials and remnant score were examined. EndoSeal MTA/SCT showed significantly shorter time of filling procedure. The number of void did not show significant differences between two techniques. No patients showed clinical signs during the follow-up periods. There were no significant differences between group AH-plus and EndoSeal MTA for remnant score. A certain calcium silicate-based sealer with SCT may give similar clinical efficiencies as much as continuous-wave technique using AH-plus sealer.

Establishment and characterization of patient-derived xenograft models of gastrointestinal stromal tumor resistant to standard tyrosine kinase inhibitors.

Gastrointestinal stromal tumors (GISTs) with KIT or platelet-derived growth factor receptor alpha (PDGFRa) oncogenic driver gene mutations, respond to tyrosine kinase inhibitors (TKIs) including imatinib, sunitinib, and regorafenib. However, most patients develop TKI resistance; therefore, novel agents are required. We established three TKI-resistant GIST patient-derived xenograft (PDX) models for effective drug development. These were PDX models harboring primary and secondary KIT and additional mutations; KIT exon 11 (p.Y570_L576del), KIT exon 17 (p.D816E), and PTEN (p.T321fs) mutations in GIST-RX1 from a patient who was unresponsive to imatinib, sunitinib, and sorafenib, and KIT exon 11 (p.K550_splice) and KIT exon 14 (p.T670I) mutations in GIST-RX2 and KIT exon 9 (p.502_503insYA) and KIT exon 17 (p.D820E) mutations in GIST-RX4 from patients with imatinib and imatinib/sunitinib resistance, respectively. The histological features and mutation statuses of GIST PDXs were consistent with those of the original patient tumors, and the models showed TKI sensitivity comparable to clinical responses. Imatinib inhibited the KIT pathway in imatinib-sensitive GIST-T1 but not GIST-RX1, RX2, and RX4. These GIST PDX models will be useful for studying TKI resistance mechanisms and evaluating novel targeted agents in GIST.

Physicochemical Properties of Epoxy Resin-Based and Bioceramic-Based Root Canal Sealers.

Three bioceramic sealers (EndoSequence BC sealer, EndoSeal MTA, and MTA Fillapex) and three epoxy resin-based sealers (AH-Plus, AD Seal, and Radic-Sealer) were tested to evaluate the physicochemical properties: flow, final setting time, radiopacity, dimensional stability, and pH change. The one-way ANOVA and Tukey's post hoc test were used to analyze the data (P = 0.05). The MTA Fillapex sealer had a highest flow and the BC Sealer presented a flow significantly lower than the others (P < 0.05). The BC Sealer and MTA Fillapex samples were not set in humid incubator condition even after one month. EndoSeal MTA had the longest setting time among the measurable materials and Radic-Sealer and AD Seal showed shorter setting time than the AH-Plus (P < 0.05). AH-Plus and EndoSeal MTA showed statistically higher values and MTA Fillapex showed statistically lower radiopacity (P < 0.05). BC Sealer showed the highest alkaline pH in all evaluation periods. Set samples of 3 epoxy resin-based sealers and EndoSeal MTA presented a significant increase of pH over experimental time for 4 weeks. In conclusion, the bioceramic sealer and epoxy resin-based sealers showed clinical acceptable physicochemical properties, but BC Sealer and MTA Fillapex were not set completely.

Prognostic impact of fibroblast growth factor receptor 2 gene amplification in patients receiving fluoropyrimidine and platinum chemotherapy for metastatic and locally advanced unresectable gastric cancers.

Although Fibroblast growth factor receptor (FGFR) 2 gene amplification and its prognostic significance have been reported in resectable gastric cancers, information on these features remains limited in the metastatic setting. The presence of FGFR2 amplification was assessed in formalin-fixed, paraffin-embedded tissues using a quantitative PCR-based gene copy number assay with advanced gastric cancer cohorts. A total of 327 patients with tumor portion of ≥70% were analyzed for clinical features. Among these patients, 260 who received first-line fluoropyrimidine and platinum chemotherapy were analyzed for survival.Sixteen of 327 patients (4.9%) exhibited FGFR2 amplification. The amplification group showed associations with age <65 years, Borrmann type 4 disease, poor performance status, poorly differentiated histology, extra-abdominal lymph node metastases, and bone metastases. The median overall survival (OS) and progression-free survival (PFS) were found to be 12.7 and 5.8 months, respectively. In univariate analysis, PFS did not differ between amplification and no amplification groups (hazard ratio [HR]=1.34, 95% confidence interval [CI]: 0.78-2.31, p=0.290), although the OS was significantly shorter in the amplification group (HR=1.92, 95% CI: 1.13-3.26, p=0.015). However, multivariate analysis indicated that FGFR2 amplification was not an independent prognostic factor for OS (HR=1.42, 95% CI: 0.77-2.61, p=0.261).Although FGFR2 amplification is associated with poorer OS, it does not appear to be an independent prognostic predictor in patients with advanced gastric cancer treated with palliative fluoropyrimidine and platinum chemotherapy.

FGFR2 Assessment in Gastric Cancer Using Quantitative Real-Time Polymerase Chain Reaction, Fluorescent In Situ Hybridization, and Immunohistochemistry.

OBJECTIVES: Fibroblast growth factor receptor 2 (FGFR2) amplification has been reported to be a target for treatment in gastric cancer. However, an optimal tissue source and method for evaluating FGFR2 have yet to be established. METHODS: Copy numbers were compared by quantitative polymerase chain reaction (qPCR) using frozen vs formalin-fixed, paraffin-embedded (FFPE) tissue and biopsy vs surgical specimens. We correlated the results of qPCR and immunohistochemistry (IHC) with fluorescence in situ hybridization (FISH) using stage IV gastric cancer biopsy specimens and validated the results in surgical specimens. RESULTS: FFPE tissues were suitable for qPCR, and biopsy specimens were equivalent to or better than surgical specimens. qPCR and IHC results exhibited an excellent correlation with FISH at eight or more copies by qPCR in any kind of tissue, 5% or more by IHC in biopsy specimens, and 10% or more by IHC in surgical specimens. FGFR2 amplification was 6.6% in stage IV gastric cancers, and 42% of these showed heterogeneous amplification and overexpression. IHC indicated a good correlation with FISH even in the heterogeneous cases. CONCLUSIONS: FFPE biopsy tissues are an adequate source for FGFR2 evaluation in gastric carcinomas, and a qPCR-based copy number assay can be used for screening. IHC is also a valid and practical method for evaluating FGFR2, considering frequent heterogeneity.

Tailored surface structure of LiFePO4/C nanofibers by phosphidation and their electrochemical superiority for lithium rechargeable batteries.

We offer a brand new strategy for enhancing Li ion transport at the surface of LiFePO4/C nanofibers through noble Li ion conducting pathways built along reduced carbon webs by phosphorus. Pristine LiFePO4/C nanofibers composed of 1-dimensional (1D) LiFePO4 nanofibers with thick carbon coating layers on the surfaces of the nanofibers were prepared by the electrospinning technique. These dense and thick carbon layers prevented not only electrolyte penetration into the inner LiFePO4 nanofibers but also facile Li ion transport at the electrode/electrolyte interface. In contrast, the existing strong interactions between the carbon and oxygen atoms on the surface of the pristine LiFePO4/C nanofibers were weakened or partly broken by the adhesion of phosphorus, thereby improving Li ion migration through the thick carbon layers on the surfaces of the LiFePO4 nanofibers. As a result, the phosphidated LiFePO4/C nanofibers have a higher initial discharge capacity and a greatly improved rate capability when compared with pristine LiFePO4/C nanofibers. Our findings of high Li ion transport induced by phosphidation can be widely applied to other carbon-coated electrode materials.

DZNep, inhibitor of S-adenosylhomocysteine hydrolase, down-regulates expression of SETDB1 H3K9me3 HMTase in human lung cancer cells.

3-Deazaneplanocin A (DZNep), an epigenetic anticancer drug, leads to the indirect suppression of S-adenosyl methionine-dependent cellular methylations by inhibiting S-adenosyl homocystein (AdoHcy) hydrolase. Although it is well known that DZNep targets the degradation of EZH2 protein, H3K27me3 HMTase, there are still uncertainties about the regulation of other types of HMTases during cell death. In this study, we describe that SETDB1 gene expression was regulated by DZNep treatment in human lung cancer cells. We confirm that DZNep induced growth inhibition and increased the dead cell population of lung cancer cells. DZNep treatment affected histone methylations, including H3K27me3 and H3K9me3, but not H3K4me3. Reduced levels of H3K27me3 and H3K9me3 were related with the decreased EZH2 and SETDB1 proteins. Real time PCR analysis showed that SETDB1 gene expression was decreased by DZNep treatment, but no effect was observed for EZH2 gene expression. We cloned the promoter region of SETDB1 and SUV39H1 genes, and performed luciferase assays. The promoter activity of SETDB1 gene was down regulated by DZNep treatment, whereas no effect on SUV39H1 promoter activity was observed. In conclusion, we suggest that DZNep regulates not only on H3K27me3 HMTase EZH2, but also H3K9 HMTase SETDB1 gene expression at the transcription level, implicating that the mechanism of action of DZNep targets multiple HMTases during the death of lung cancer cells.

Quantitative analysis of H5N1 DNA hybridization on nanowell array electrode.

A nanowell array electrode-based electrochemical quantitative system without amplification was developed and applied for the detection of H5N1 target DNA. An 18-mer probe was immobilized on a nanowell array electrode with a diameter of 500 nm, which was coated with streptavidin and a self-assembly monolayer (SAM). The surface properties of probe DNA hybridization with complementary target DNA were characterized using atomic force microscopy (AFM) and electrochemical impedance spectroscopy (EIS). The AFM image shows that the depth of nanowell was reduced from 200 nm to 15 nm due to the formation of a DNA hybridization complex on the streptavidin/SAM structure. Differences in charge transfer resistance (deltaR(ct)) in EIS upon hybridization of the probe DNA with complementary target DNA were analyzed and used for the quantitation of H5N1 DNA. This approach shows that the quantitative analysis of H5N1 DNA ranging from 1 pM to 1 microM DNA is possible on a nanowell array electrode.

S-adenosyl methionine specifically protects the anticancer effect of 5-FU via DNMTs expression in human A549 lung cancer cells.

Cellular methylation is associated with stabilization of the chromatin structure. S-adenosyl methionine (SAM), a metabolite of methionine metabolism, is the methyl donor of essential cellular methyltransferase reactions. Using 3-(4,5-dimethylthiazol-2-yl)-2,5-dephenyl tetrazolium bromide (MTT) assay, we found that combination treatment of SAM and 5-fluorouracil (5-FU) specifically protected the anticancer effect of 5-FU, whereas the combination of SAM and cisplatin had no effect. This result was confirmed by FACS analysis. The combination treatment of SAM and 5-FU significantly decreased the dead cell population, while the G1 cell population was slightly increased, suggesting that protection of SAM is not associated with the cell cycle arrest of DNA-damaging drugs. We also analyzed which cellular methylation-related proteins were involved in the protective effect. Results showed the expression of DNA methyltransferases (DNMTs) was decreased with 5-FU alone but was increased with the combination treatment of SAM and 5-FU, suggesting that SAM protects the anticancer effect of 5-FU by regulating the expression of DNMTs. Taken together, the results indicated that SAM specifically modulates the anti-cancer effect of the DNA damage agent 5-FU and this may be modulated by aberrant DNA methylation.

Bronchial hyperresponsiveness in preschool children with allergic rhinitis.

BACKGROUND: Nonasthmatic subjects with allergic rhinitis often have bronchial hyperresponsiveness (BHR), characteristic of asthma. The presence and degree of atopy is suggested to be important for BHR in patients with asthma. We aimed to assess BHR to methacholine (direct stimulus) and to adenosine 5'-monophosphate (AMP; indirect stimulus) in preschool children with allergic rhinitis and to investigate their relationship with the degree of atopy. METHODS: Methacholine and AMP bronchial challenges were performed in preschool children with allergic rhinitis (n = 96), using a modified auscultation method. The end point concentration, resulting in audible wheezing and/or oxygen desaturation, was determined for each challenge. The degree of atopy was assessed using serum total IgE levels, the number of positive skin-prick tests, and atopic scores (sum of graded wheal size). RESULTS: BHR to methacholine (end point concentration, ≤8 mg/mL) and to AMP (end point concentration, ≤200 mg/mL) was observed in 32 (33.3%) and 26 (27.1%) subjects, respectively. No significant relationship was observed between BHR to methacholine and any atopy parameter. In contrast, the atopic scores were higher in the AMP-BHR(+) group compared with the AMP-BHR(-) group, and a significant association was found between the degree of atopic scores and the frequency of BHR to AMP (score for trend, p = 0.006). Such a relationship was not observed for serum total IgE levels and the number of positive SPTs. CONCLUSION: BHR to methacholine and BHR to AMP were detected in a significant proportion of preschool children with allergic rhinitis. The degree of atopy in terms of atopic scores seems to be an important factor for BHR to AMP but not for BHR to methacholine.

Airway refractoriness to inhaled mannitol after repeated challenge.

Exercise and inhaled mannitol are thought to cause bronchoconstriction through a similar mechanism in asthma. The response to exercise becomes refractory with repeated challenges. This study aimed to investigate whether repeated challenge with mannitol induces refractoriness, as with exercise. Forty-one children with asthma underwent two consecutive dose-response mannitol challenges (Phase 1); the second challenge proceeded after recovery (FEV(1) : 95% or more of baseline value) from the first. The response to mannitol was expressed as a provocative dose causing a 15% fall in FEV(1) (PD(15) ) and the response-dose ratio (RDR) (% fall in FEV(1) /cumulative dose). In 18 subjects who were deemed to have mannitol refractoriness in Phase 1, a mannitol challenge was performed before and after a methacholine challenge (Phase 2). In Phase 1, the time taken for the FEV(1) to recover after the first mannitol challenge ranged from 20 to 100 min with a median of 50 min. In the 23 subjects with a measurable mannitol PD(15) in both challenges, the geometric mean (95%CI) PD(15) in the second challenge (163 mg [114-232]) was significantly higher than that in the first challenge (66 mg [50-88], P < 0.001). The geometric mean (95%CI) RDR decreased from the value of 0.083%/mg (0.055-0.125) in the first challenge to 0.029%/mg (0.017-0.048) in the second challenge (P < 0.001). In Phase 2, prior challenge with methacholine or mannitol did not significantly alter subsequent bronchoconstriction to the opposite challenge. Repeated challenge with mannitol resulted in less bronchoconstriction when compared with the initial challenge. This refractoriness seems not to be attributable to functional loss of responsiveness or non-specific effect of prior bronchoconstriction.

Relationships of methacholine and adenosine 5'-monophosphate (AMP) responsiveness to the postbronchodilator FEV1/FVC ratio in children with asthma.

BACKGROUND: Airway remodeling has been assumed to cause bronchial hyperresponsiveness (BHR). A low postbronchodilator FEV1/FVC ratio has been suggested to be a functional surrogate marker of airway remodeling in asthma. BHR is commonly assessed by bronchial challenges using direct or indirect stimuli. OBJECTIVE: The aim of this study was to compare BHR to methacholine and adenosine 5'-monophosphate (AMP) with regard to their relationship with a marker of airway remodeling in children with asthma. METHODS: Methacholine and AMP challenge tests were performed in 129 children with asthma, aged 12 years, and a provocative concentration causing a 20% fall in FEV1 (PC20) was calculated for each challenge. All subjects also underwent pre- and postbronchodilator spirometry. A postbronchodilator FEV1/FVC ratio below the lower limits of normal was used as a marker of airway remodeling. RESULTS: A low postbronchodilator FEV1/FVC ratio was found in 17 subjects (13.2%). These subjects had a significantly lower methacholine PC20 (geometric mean: 0.63 mg/mL, range of 1 SD: 0.17-2.29) than those (n = 112) with a normal postbronchodilator FEV1/FVC ratio (2.42 mg/mL, 0.57-10.32, p = .000), whereas AMP PC20 was similar between the two groups (22.1 mg/mL, 3.9-125.9 vs. 27.7 mg/mL, 4.2-183.5, p = .231). In the whole group of subjects, methacholine PC20, but not AMP PC20, correlated significantly with the postbronchodilator FEV1/FVC ratio (r = 0.340, p = .000, and r = 0.056, p = .526, respectively). CONCLUSIONS: Our results provide evidence, though indirect, that BHR to methacholine is related to airway remodeling in children with asthma and suggest that BHR to methacholine may be a better marker of airway remodeling than BHR to AMP.

Bronchial hyperresponsiveness to methacholine and adenosine monophosphate and the degree of atopy in children with allergic rhinitis.

BACKGROUND: nonasthmatic patients with allergic rhinitis often have bronchial hyperresponsiveness (BHR). Not only the presence but also the degree of atopy are important factors in BHR of patients with asthma. BHR is commonly evaluated by bronchial challenges using direct or indirect stimuli. OBJECTIVES: to assess BHR to methacholine (direct) and to adenosine monophosphate (AMP) (indirect) in children with allergic rhinitis and to compare their relationships with the degree of atopy. METHODS: methacholine and AMP challenges were performed in 88 children with allergic rhinitis, and a provocative concentration causing a 20% decrease in forced expiratory volume in 1 second (PC(20)) was calculated for each challenge. The degree of atopy was measured using serum total IgE levels, number of positive skin prick test results, and atopic scores (sum of graded wheal size). RESULTS: BHR to methacholine (PC(20) <8 mg/mL) and to AMP (PC(20) <200 mg/mL) was observed in 22 (25%) and 30 (34%) patients, respectively. No association was found between BHR to methacholine and any atopy parameter. In contrast, serum total IgE levels and atopic scores were higher in the group with BHR to AMP than in the group without BHR to AMP. Furthermore, a significant association was found between the degree of these 2 parameters and BHR to AMP (score for trend, P < .001 and P = .03, respectively). CONCLUSIONS: both BHR to methacholine and BHR to AMP were detected in a significant proportion of children with allergic rhinitis. The degree of atopy seems to be an important factor in BHR to AMP but not in BHR to methacholine.

The role of inhaled and/or nasal corticosteroids on the bronchodilator response.

PURPOSE: To compare the profiles of the bronchodilator response (BDR) among children with asthma and/or allergic rhinitis (AR) and to determine whether BDR in these children is reduced by treatment with inhaled and/or nasal corticosteroid. METHODS: Sixty-eight children with asthma (mean age, 10.9 years), 45 children with comorbid asthma and AR (mean age, 10.5 years), and 44 children with AR alone (mean age, 10.2 years) were investigated. After a 2-week baseline period, all children were treated with inhaled fluticasone propionate (either 100 or 250 µg b.i.d., tailored to asthma severity) or nasal fluticasone propionate (one spray b.i.d. in each nostril) or both, according to the condition. Before and 2 weeks after starting treatment, all children were evaluated with spirometry and bronchodilator testing. BDR was calculated as a percent change from the forced expiratory volume in 1 second (FEV(1)) at baseline. RESULTS: The mean BDR was 10.3% [95% confidence interval (CI) 8.3-12.4%] in children with asthma, 9.0% (95% CI 7.3-10.9%) in subjects with asthma and AR, and 5.0% (95% CI 4.1-5.9%) in children with AR alone (P<0.001). After treatment, the mean BDR was reduced to 5.2% (95% CI 4.2-6.3%) (P<0.001) in children with asthma and to 4.5% (95% CI 3.5-5.5%) (P<0.001) in children with asthma and AR. However, children with rhinitis showed no significant change in BDR after treatment, with the mean value being 4.7% (95% CI 3.7-5.8%) (P=0.597). CONCLUSION: The findings of this study imply that an elevated BDR in children with AR cannot be attributed to nasal inflammation alone and highlights the close relationship between the upper and lower airways.