Measurement of Anions in Tear Fluid Using Ion Chromatography.

PURPOSE: Tear fluid (TF) contains a variety of electrolytes that exhibit a strong correlation with its osmotic pressure. These electrolytes are also related to the etiology of diseases on ocular surfaces such as dry eye syndromes and keratopathy. Although positive ions (cations) in TF have been investigated to understand their roles, negative ions (anions) have hardly been studied because applicable analytical methods are restricted to a few kinds. In this study, we established a method to analyze the anions involved in a sufficiently small amount of TF for in situ diagnosis of a single subject. METHODS: Twenty healthy volunteers (10 men and 10 women) were recruited. Anions in their TF were measured on a commercial ion chromatograph (IC-2010, Tosoh, Japan). Tear fluid (5 µL or more) was collected from each subject with a glass capillary, diluted with 300 µL of pure water, and conveyed to the chromatograph. We successfully monitored the concentrations of bromide, nitrate, phosphate, and sulfate anions (Br - , NO 3- , HPO 42- , and SO 42- , respectively) in TF. RESULTS: Br - and SO 42- were universally detected in all samples, whereas NO 3- was found in 35.0% and HPO 42- in 30.0% of them. The mean concentrations (mg/L) of each anion were Br - , 4.69 ± 0.96; NO 3- , 0.80 ± 0.68; HPO 42- , 17.48 ± 7.60; and SO 42- , 3.34 ± 2.54. As for SO 42- , no sex differences or diurnal variations were observed. CONCLUSIONS: We established an efficient protocol to quantitate various inorganic anions involved in a small amount of TF using a commercially available instrument. This is the first step to elucidate the role of anions in TF.

Upregulation of REV7 correlates with progression of malignant melanoma.

REV7 is a multifunctional protein implicated in DNA damage tolerance, cell cycle control, and gene expression, and is involved in the carcinogenesis of various human tumors. It has been reported that REV7 expression is associated with ultraviolet-induced mutagenesis; however, the role of REV7 expression in skin cancers, including malignant melanomas, remains unclear. In the present study, we investigated the clinical and biological significance of REV7 in malignant melanoma. Levels of REV7 expression in human skin cancers were evaluated immunohistochemically. Positive expression of REV7 was frequently observed in malignant melanomas, as well as in squamous cell carcinomas and basal cell carcinomas. Enhanced immunoreactivity to REV7 was closely linked with cell proliferation assessed by Ki-67 labeling indexes in the three skin cancers, and was related with tumor thickness in malignant melanomas. REV7 depletion in malignant melanoma cells MEWO and G361 suppressed cell proliferation, migration, and invasion abilities. REV7 depletion also affected the expression of intracellular signaling molecules AKT and ERK in MEWO cells, resulting in downregulation of ERK signal activation. In addition, REV7 depletion facilitated sensitivity to cisplatin, but not to dacarbazine, in MEWO cells. Our results suggest that REV7 expression correlates with disease progression of malignant melanoma.

Analysis of pharmacist-patient communication using the Roter Method of Interaction Process Analysis System.

BACKGROUND: Effective communication between pharmacists and patients is a crucial factor in ensuring that medications are used properly. However, few studies have examined the contents of actual on-site communications between pharmacists and patients. OBJECTIVE: To identify the characteristics of and problems with routine communications between pharmacists and patients using the Roter Method of Interaction Process Analysis System (RIAS). METHODS: Conversations between pharmacists and simulated patients (SPs) were recorded and transcribed. Using the RIAS technique, their utterances were classified into 42 categories, and these were further divided into 11 clusters, such as open- and closed-ended questions, and analyzed. Furthermore, the influence that the different scenarios performed by the pharmacists may have had on the structure of their communication was investigated. All of the transcripts were double-coded by two certified coders. RESULTS: A total of 57 pharmacists took part in the study. The mean ratio of utterances made by SPs and pharmacists were 44% and 56%, respectively. The percentage of pharmacists' questions was more than double that of SPs' for both open- and closed-ended questions. In the influence that the different scenarios, the scenarios for patients with cancer was significantly higher ratio of utterances by the pharmacists. CONCLUSIONS: Pharmacists' communications tended to focus more on information-gathering activities that concentrated on closed-ended questions and frequent counseling, or directing utterances about the medication than on considering the patient's background. On the other hand, the pharmacists did communicate in ways that matched each patient's disease. This study identified the structure of pharmacists' on-site communications, and revealed the associated characteristics and problems.

Dietary ß-carotene regulates interleukin-1ß-induced expression of apolipoprotein E in astrocytes isolated from stroke-prone spontaneously hypertensive rats.

Stroke-prone spontaneously hypertensive rats (SHRSP) have an abnormality in cholesterol synthesis, but the pathological relevance of this to stroke and related neuronal disorders is not yet clear. The induction of astrocyte-derived cholesterol transportation to neurons by apolipoprotein E (apoE) promotes neuronal repair after brain injuries such as stroke. Such repair is reduced by interleukin-1 beta (IL-1ß) and stroke conditions. Furthermore, fibroblast growth factor 1 (FGF1) regulates the production of apoE-cholesterol-rich high density lipoproteins (HDL) and induces gliosis of astrocytes. On the other hand, high levels of plasma carotenoids reduce the risk of ischemic stroke. Thus, we investigated the expression of apoE in primary astrocytes that had been treated with IL-1ß or ß-carotene. In addition, we compared the expression levels of Apoe genes in astrocytes from SHRSP/Izm and normal control rats, Wistar-Kyoto rats (WKY/Izm) following hypoxia/reoxygenation (H/R). The expression levels of genes and proteins were investigated by RT-PCR, Western blotting (WB), and immunofluorescence analysis. IL-1ß decreased the expression levels of the Apoe gene. Conversely, ß-carotene significantly enhanced the expression levels of genes related to cholesterol regulation, including Abca1, Abcg1, Hmgcr as well as Apoe. During H/R, the gene expression levels of Apoe were decreased in the SHRSP/Izm rats in comparison with the WKY/Izm rats. These results suggest that IL-1ß decreases Apoe expression levels, whereas ß-carotene strongly elevates Apoe levels and inhibits FGF1-mediated gliosis of astrocytes. Furthermore, under hypoxic stress, astrocytes isolated from SHRSP/Izm rats displayed altered regulation of Apoe compared with those from WKY/Izm rats.