Factors influencing subjective symptoms in dry eye disease

AIM: To retrospectively investigate the association between dry eye symptoms and clinical or in vivo confocal microscopy parameters in patients with dry eye disease (DED), and to compare these parameters between eyes with DED and normal subjects. METHODS: This retrospective, cross-sectional, controlled study comprised 25 consecutive patients with non-Sjögren dry eye disease and age- and sex-matched 25 healthy subjects. Each patient underwent a complete examination of the ocular surface in the following order: tear osmolarity measurements, InflammaDry test, tear break-up time, corneal fluorescein staining, Schirmer I test, subjective symptoms questionnaire using the dry eye-related quality-of-life score (DEQS), and in vivo confocal microscopy analysis of the central cornea. Beck depression inventory (BDI) as depressive scale and history of medications and smoking were also evaluated. Stepwise multiple regression analysis was used to assess the factors affecting the DEQS. RESULTS: In univariate analysis, DEQS was associated with tear break-up time (ρ=-0.48, P=0.01), oral medications, such as hypotensive drug (ρ=0.56, P=0.004) and anti-depressant (ρ=0.57, P=0.003), and BDI (ρ=0.61, P=0.001) in patients with DED. In multiple regression analysis, explanatory variables relevant to the DEQS were the anti-depressant medications (P=0.04, partial regression coefficient B=21.04) and BDI (P=0.02, B=0.76, adjusted R2=0.54) in these patients. CONCLUSION: Our study shows a significant association between depression and dry eye symptoms. It suggests that dry eye symptoms associate with higher depressive symptoms and its medications, although our patients were not followed longitudinally

Fluid dynamics in patients with nasal disease

Computational fluid dynamics （CFD） analysis is useful for quantitative assessment in patients with upper airway obstructions. We compared CFD analysis with rhinomanometry （RM） and acoustic rhinometry （AR）. Twenty patients with nasal and paranasal diseases who required computed tomography assessment underwent RM and AR. We measured the pressure and velocity at four parts of the upper airway using CFD analysis. Then we evaluated the correlation among CFD analysis, RM, and AR. CFD analysis detected obstruction sites in the nasal airway and pharynx in 14 and 2patients, respectively. High negative pressure accompanied the nasal obstruction, even behind the nasal cavity. Nasal airway pressure measured using CFD analysis strongly correlated with nasal resistance in RM （Spearman correlation coefficient＝0.853）. CFD analysis’s sensitivity and specificity to detect the obstruction were 84.6％ and 57.1％, respectively （compared to those of RM） and 83.3％ and 50.0％, respectively （compared to those of AR）. The CFD analysis’s ability to detect obstruction was comparable to that of RM and AR; therefore, it may help evaluate the upper airways in patients with nasal and paranasal diseases. We found impaired nasal ventilation also affected other parts of the upper airway. Further studies with a larger sample size are required to validate the use of CFD analysis for assessing the degree of upper airway ventilation disorders

Computational fluid dynamics analysis in patients with nasal disease

Computational fluid dynamics （CFD） analysis is useful for quantitative assessment in patients with upper airway obstructions. We compared CFD analysis with rhinomanometry （RM） and acoustic rhinometry （AR）. Twenty patients with nasal and paranasal diseases who required computed tomography assessment underwent RM and AR. We measured the pressure and velocity at four parts of the upper airway using CFD analysis. Then we evaluated the correlation among CFD analysis, RM, and AR. CFD analysis detected obstruction sites in the nasal airway and pharynx in 14 and 2patients, respectively. High negative pressure accompanied the nasal obstruction, even behind the nasal cavity. Nasal airway pressure measured using CFD analysis strongly correlated with nasal resistance in RM （Spearman correlation coefficient＝0.853）. CFD analysis’s sensitivity and specificity to detect the obstruction were 84.6％ and 57.1％, respectively （compared to those of RM） and 83.3％ and 50.0％, respectively （compared to those of AR）. The CFD analysis’s ability to detect obstruction was comparable to that of RM and AR; therefore, it may help evaluate the upper airways in patients with nasal and paranasal diseases. We found impaired nasal ventilation also affected other parts of the upper airway. Further studies with a larger sample size are required to validate the use of CFD analysis for assessing the degree of upper airway ventilation disorders

Quantification of (-)-epigallocatechin-3-gallate Inhibition of Migration and Invasion of Oral Squamous Cell Carcinoma Cell Lines Using Real-time Cell Analysis

Catechins found in green tea, in particular (−)-epigallocatechin-3-gallate (EGCG), have antitumor activity. The primary antitumor actions of catechins are anti-oxidative, anti-angiogenic, and anti-metastatic effects. Cell migration and invasion contribute to the metastatic potential of tumors. Real-time cell analysis (RTCA) measures cell migration and invasion in vitro. In the present study, using RTCA, we investigated whether the cell migration and invasion of oral squamous cell carcinomas (OSCCs) of the tongue and floor of the mouth were inhibited by EGCG. Studies were performed using the human SCC-4 and SAS cell lines, which are poorly differentiated OSCCs of the tongue, and the HO-1-u-1 cell line, an OSCC of the floor of the mouth. SCC-4 cells exhibited high cell migration and invasion compared with the SAS and HO-1-u-1 cells. EGCG was most effective in inhibiting the migration and invasion of SCC-4 cells, and inhibited OSCC cell invasion more strongly than it inhibited cell migration. EGCG inhibited the expression of matrix metalloproteinase (MMP)-2, MMP-9, and integrin α1 and β1 mRNA in the OSCC cell lines, particularly SCC-4 cells. The findings of the present study suggest that EGCG inhibits OSCC cell migration and invasion by inhibiting MMP-2, MMP-9, and integrin α1and β1 expression. Thus, EGCG may be a suitable agent or lead compound for the inhibition of OSCC metastasis

Investigation of Cell Migration and Invasion Using Real-time Cell Analysis, as well as the Association with Matrix Metalloproteinase-9 in Oral Squamous Cell Carcinomas

The recently developed technology of real-time cell analysis (RTCA) was designed to analyze cell migration and invasion in vitro. In this study, we investigated these cellular factors in oral squamous cell carcinomas (OSCCs) of the tongue and floor of the mouth with RTCA. We also examined the associated matrix metalloproteinases (MMPs) and integrins. We used the cell lines SCC-4 and SAS, which are human poorly differentiated OSCCs from the tongue, and HO-1-u-1, which are human poorly differentiated OSCCs from the floor of the mouth. Using RTCA, cell migration was assessed on fibronectin–coated CIM-Plates, and invasion was assessed on fibronectin- and matrigel-coated CIM-Plates. SCC-4 cells demonstrated a high ability for cell migration and invasion compared with SAS and HO-1-u-1 cells. The SCC-4 cells also expressed high levels of MMP-9 and integrin α1 mRNA compared with SAS and HO-1-u-1 cells. The MMP inhibitor Marimastat blocked migration and invasion of all OSCCs. The findings suggest that MMP-9 is associated with cell migration and invasion in OSCCs, and indicate that RTCA will be useful for analyzing the metastatic capability of OSCCs and developing more effective new drugs for this disease

Ethanol-induced Stress Leads to Apoptosls Via Endoplasmic Reticulum Stress in SK-Hepl Cells

Alcoholic liver disease causes oxidative stress and induces apoptosis during alcohol metabolism. Ethanol causes endoplasmic reticulum (ER) stress in hepatocytes, stimulating the unfolded protein response (UPR) pathway and/or Ca2+-dependent calpain and caspase-4 activities. However, it is poorly understood whether ethanol-induced oxidative stress directly leads to apoptosis promoted by ER stress-associated pathways. This study investigated this question in human liver adenocarcinoma (SK-Hep1) cells, which were treated with 200 mM ethanol for 5 hours in the presence or absence of the antioxidant N-acetyl-cysteine (NAC). We found that treatment with ethanol significantly increased ROS production and cellular apoptosis in the SK-Hep1 cells, and that this response was significantly suppressed by pretreatment with NAC. Furthermore, pretreatment with NAC significantly reduced the observed increases in the mRNA expressions of Bip, Chop, and sXbp-1, and the activity of caspase-3 in ethanol-induced apoptotic cells. However, pretreatment with NAC did not attenuate the transient rise in cytosolic Ca2+ nor the activities of caspase-4 and calpain induced by ethanol. Together, these results revealed that ethanol-induced stress promotes apoptosis not only through mitochondria-mediated pathways, but also via ER stress. The findings further suggested that ethanol-induced oxidative stress and non-oxidative stress both stimulate the pathway regulating ER stress-mediated apoptosis

Quantification of (–)-Epigallocatechin-3-gallate Inhibition of Anaplastic Thyroid Cancer Cell Line Adhesion and Proliferation Using Real-time Cell Analysis

Anaplastic thyroid cancer (ATC) has a poor prognosis because of immediate metastasis. Several studies in humans and animals have suggested that the ingestion of green tea or its active ingredient (–)-epigallocatechin-3-gallate (EGCG) may decrease the risk of cancer. Using a recently developed real-time cell analysis (RTCA) system, we have shown previously that EGCG inhibits cell migration and the invasion of oral cavity cancers by suppressing matrix metalloproteinases. In the present study we used RTCA to investigate the effects of EGCG on cell adhesion to fibronectin-coated plates using three cancer cell lines: one ATC cell line (TCO-1) and two poorly differentiated oral squamous cell carcinomas (OSCCs) cell lines (SAS and HO-1-u-1; originating from the tongue and floor of the mouth, respectively). EGCG (50µM) inhibited the adhesion of all three cell lines. In addition to its effects on cell adhesion, 50µM EGCG inhibited the cell proliferation of TCO-1 cells. Furthermore, EGCG decreased αV integrin (ITGAV) mRNA levels in all three cell lines, suggesting that EGCG inhibits the cell adhesion and proliferation of OSCC and ATC cells via suppression of integrin expression. Therefore, EGCG represents a useful dietary constituent or a lead compound for counteracting metastasis of oral cavity cancers and thyroid cancers

Quantification of Cell Migration and Invasion, and Their Association with Periostin in Anaplastic Thyroid Cancer, Using a Real-time Cell Analyzer

Anaplastic thyroid cancer (ATC) is known to be a highly malignant cancer of the thyroid with a high mortality rate. In a previous study, we used real-time cell analysis (RTCA) to analyze cell migration and invasion of oral squamous cell carcinomas (OSCCs) of the tongue and floor of the mouth. In the present study, we investigated cell migration and invasion of ATC using RTCA, as well as their association with periostin, matrix metalloproteinases (MMPs), and integrins. Experiments were performed on TCO-1 and HTC/C3 cells, which are human ATC cell lines. OSCC cell lines were used for comparison. Using the cell analysis system, cell migration was assessed on fibronectin-coated CIM-Plates, whereas invasion was assessed on fibronectin- and matrigel-coated CIM-Plates. SCC-4 cells exhibited high cell migration and invasion activity compared with other OSCC cell lines. TCO-1 cells exhibited equivalent cell invasion but stronger migration than SCC-4 cells. Although TCO-1 cells had strong invasive activity, they did not express MMP-9, unlike SCC-4 cells. Conversely, periostin expression was high in TCO-1 cells. Therefore, periostin expression appears to be associated with the cell migration and invasion activity of ATC. The RTCA system will be useful for the analysis of the metastatic characteristics of ATC in head and neck cancer

Increase in Matrix Metalloproteinase-2 and 9 in the Liver of Nonalcoholic Steatohepatitis (NASH) Model Rats

Nonalcoholic steatohepatitis (NASH) is regarded as a hepatic manifestation of the metabolic syndrome which can progress to hepatic cirrhosis and hepatocellular carcinoma. It is thought that matrix metalloproteinases (MMPs) play an important role in hepatic fibrosis and we previously reported a correlation between oxidative stress and MMP-9 expression. However, the expression of MMPs and tissue inhibitors of metalloproteinases (TIMPs) in the progression of NASH is unclear. In this study we used spontaneously hypertensive and hyperlipidemic rats (SHHR) fed a high-fat diet and 30% sucrose solution (HFDS) as a model for NASH, in order to clarify the relationships between oxidative stress, liver weight (LW), MMPs and TIMPs at various time-points in the progression of NASH. Male SHHR and Sprague-Dawley (SD) rats were divided into four groups: SHHR-normal diet (ND), SHHR-HFDS, SD-ND and SD-HFDS. Hepatic fibrosis was clearly increased at 13 months in SHHR-HFDS, resembpling NASH. LW and oxidative stress markers in plasma were increased in SHHR-HFDS compared to the other groups. Oxidative stress was correlated with LW in all rats. Expression of MMP-2, MMP-9, TIMP-1 and TIMP-2 mRNA, measured by real-time polymerase chain reaction, was increased in the liver of SHHR-HFDS at 13 months. This study suggests that oxidative stress, MMPs and TIMPs may play an important role in the progression of NASH

Significance of Coronary Artery Calcium Score in the Target Lesion Evaluated by Multi-detector Computed Tomography for Selecting Treatment of Rotational Atherectomy in Patients with Coronary Artery Disease

We investigated whether coronary artery calcium score (CAC) in the target lesion on the multidetector computed tomography angiography (CTA) predicts the addition of the Rotational atherectomy (Rota) during percutaneous coronary intervention (PCI). Lesion CAC on CTA were evaluated with quantitative coronary analysis (QCA) on coronary angiography for predicting the Rota treatment in 114 consecutive patients (165 target lesions) with first PCI (68 ± 9 years old, females: 17.6%). Rota was added in 8 patients (11 lesions). The lesion length and diameter stenosis on QCA, and lesion length and lesion CAC on CTA were the primary factors associated with the addition of Rota. Using the cut-off value based on receiver operating characteristic analysis, the sensitivity and specificity for predicting the Rota based on QCA was 72.7% in 8 of 11 lesions (vessels) with Rota and the specificity was 74% in 114 of 154 without Rota in the lesion length of ≥ 23mm (χ2=10.9, p=0.001), and 54.5% in 6 of 11 lesions with Rota and the specificity was 79.2% in 122 of 154 without Rota in the diameter stenosis of ≥ 83% (χ2=6.6, p=0.01). Those based on CTA were 90.9% in 10 of 11 lesions with Rota and 77.3% in 119 of 154 without Rota in the lesion length of ≥ 34mm (χ2=24.1, p<0.001), and 90.9% in 10 of 11 with Rota and 88.3% in 136 of 154 without Rota in the lesions with CAC ≥453 (χ2=45.7, p<0.001). Lesion CAC on CTA is most predictive of addition of Rota during PCI