A comparative proteomic study of secretomes in kaempferitrin-treated CTX TNA2 astrocytic cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Kaempferitrin is extracted in significantly high quantities from the leaves of Cinnamomum osmophloeum (C.O) and Bauhinia forficata, and are used as an antidiabetic herbal remedy in China and Brazil. Commercial product using dry Cinnamomum osmophloeum leaves has been sold locally in Taiwan. Oral administration of kaempferitrin reduced blood sugar in diabetic rats. AIM OF THE STUDY: Though previously demonstrated to activate the classical insulin signaling pathways, a mechanism for kaempferitrin is still not fully understood. Also, studies on kaempferitrin on immune related cells have been inconclusive, and people consuming extract containing kaempferitrin often happen to be at high risk of diabetes and neurodegenerative diseases. Therefore, for kaempferitrin to be used every day, a comprehensive study is needed. MATERIALS AND METHODS: Astrocytic cell line was used as a model to test the differentially regulated secretomes, to test kaempferitrin effect on CNS glia, on pro-inflammatory cytokines, and to test how different the mechanism of kaempferitrin is from that of insulin. CTX TNA2 astrocytic cells were differentially treated with and without 10 µM kaempferitrin for 24 h, and the conditioned medium was collected. For the proteomic study, protein in conditioned medium was trypsin digested, and resulting peptides in kaempferitrin/non-treated sample pair were differentially dimethyl labeled. The labeled peptides were further fractionated by StageTip-based strong-exchange method before LC-MS/MS analyses. Levels of interesting proteins were verified using Western or Eliza. C.O. leaf crude extract treated samples were included for a comparison of effects of purified kaempferitrin vs. kaempferitrin containing crude extract. RESULTS AND CONCLUSIONS: Data were obtained via ProteomeXchange with identifier PXD002814. It was found that no pro-inflammatory cytokines or inhibitory ECM were elevated upon treatment of kaempferitrin or a crude extract of C.O. leaves. This suggests that prolonged use of kaempferitrin containing herbs may not increase pro-inflammatory reaction. LDL-R trafficking between the cell membrane and the extracellular niche was regulated by kaempferitrin toward reduced secretion. Our proteomic study also demonstrated that molecules related to plasma membrane recycling were regulated by kaempferitrin. Our discoveries provide evidence that link kaempferitrin regulation for LDL-R and membrane recycling to the blood lipid regulation by the C.O. leaves extract. However, these proteins were differently regulated when cells were treated with crude extract. This demonstrates that the molecular interactions within crude extract of herbs are complex and may not act similar to the compound purified from the crude extract.

The influence of orthokeratology on intraocular pressure measurements.

PURPOSE: Underestimation of IOP in a myopic patient may lead to misjudgment of the risk of glaucoma. This study investigated the influence of orthokeratology-induced change in CCT on IOP measured by the non-contact pneumotonometer (NCT), Goldmann applanation tonometer (GAT), and Pascal dynamic contour tonometer (PDCT). METHODS: This study was conducted to examine the eyes of 34 patients who received orthokeratology for myopia. CCT and IOP were measured, and IOP was obtained with the NCT, GAT, and PDCT. The associations between changes in measured IOP and change in CCT at different orthokeratology follow-up time points were evaluated by linear regression analysis. RESULTS: Change in IOP measured by the three tonometries correlated significantly with change in CCT after one-week application of orthokeratology. The correlation was strongest for NCT followed by GAT and PDCT. The changes in measured IOP corresponding to a 10 µm decrease in CCT were 0.7-0.9, 0.4-0.6, and 0.2-0.3 mm Hg for NCT, GAT, and PDCT, respectively. CONCLUSIONS: The IOP measured by the three methods--NCT, GAT, and PDCT--decreased as a result of the change in CCT induced by orthokeratology. The influence on NCT and GAT was greater than that on PDCT.

Association between the characteristics of metabolic syndrome and Alzheimer's disease.

Various epidemiological studies have shown that type 2 diabetes and metabolic syndrome are highly correlated with Alzheimer's disease (AD). Here, we sought to assess the impact of metabolic syndrome characteristics on the progression of AD. Five-week-old male, spontaneously hypertensive (n = 32) and Wistar Kyoto (abbreviated WKY; n = 8) rats were divided into 5 groups (each n = 8): WKY, hypertension (HTN), streptozotocin-induced diabetes (STZ), high-fat diet (HFD), and STZ + high-fat diet-induced diabetes mellitus (DM). All animals were sacrificed and samples of the blood, liver, and brain were collected for further biological analysis. During the 15-week period of induction, the STZ and DM groups (animals injected with low-dose STZ) had significantly higher fasting glucose levels; the HFD group had elevated insulin levels, but normal blood glucose levels. The HFD and DM groups had hypercholesterolemia and higher hepatic levels of triglycerides and cholesterol. Additionally, correlations between HFD and elevated brain amyloid-beta 42 (Aß-42), hyperglycemia and down-regulation of brain insulin receptor, and serum Aß-42 and hepatic triglyceride concentrations (r(2) = 0.41, p < 0.05) were observed. Serum C-reactive protein and malondialdehyde did not appear to have a significant influence on the association with biomarkers of AD. Thus, our study demonstrated that rats with characteristics of metabolic syndrome had a large number of biomarkers predicting AD; however, no relationship between traditional inflammatory and oxidative markers and AD was found. Further studies are necessary to prove that these findings in rats are relevant to AD processes in humans.

Relationship between air pollution and outpatient visits for nonspecific conjunctivitis.

PURPOSE: Past studies present evidence of associations between air pollution and human ocular symptoms; however, to the knowledge of the authors, research investigating the hazardous effects of air pollution on nonspecific conjunctivitis is nonexistent. This study investigates the relationship between air pollution and outpatient visits for nonspecific conjunctivitis in Taiwan. A multiarea analysis was conducted to examine and assess the risks of short-term effects of particulate matter (PM), nitrogen dioxide (NO2), sulfur dioxide (SO2), ozone (O3), and carbon monoxide on nonspecific conjunctivitis. METHODS: Data were collected from outpatient visits for nonspecific conjunctivitis from seven air-quality-monitoring areas. To find immediate and lag effects of air pollution, an area-specific, case-crossover analysis was performed and a meta-analysis with random effects was used to combine the area-specific RESULTS: Results. The effects on outpatient visits for nonspecific conjunctivitis are strongest for O3 and NO2, with a 2.5% increase (95% confidence interval [CI], 0.9-4.1) for a 16.4 ppb (parts per billion) concentration rise in O3 and a 2.3% increase (95% CI, 0.7-3.9) for an 11.47 ppb concentration rise in NO2. Effects are also found for particulate matter with an aerodynamic diameter ≤ 10 µm (PM10) and SO2. Effects are more prominent in winter because the analysis was stratified according to season. CONCLUSIONS: The air pollutants NO2, SO2, O3, and PM10 increase the chances of outpatient visits for nonspecific conjunctivitis and have no evident lag effects.

Preparation of immunogens and production of antibodies.

The quality of reagents greatly affects the interpretation of serological tests. Methods used in conventional viral purification and molecular cloning and expression of target viral proteins to obtain antigens for immunization are presented. Immunization of rabbits, mice and chickens and isolation of immunoglobulin from immunized animals also are described.

Effectiveness and stability of heterologous proteins expressed in plants by Turnip mosaic virus vector at five different insertion sites.

The N-terminal (NT) regions of particular protein-coding sequences are generally used for in-frame insertion of heterologous open reading frames (ORFs) in potyviral vectors for protein expression in plants. An infectious cDNA clone of Turnip mosaic virus (TuMV) isolate YC5 was engineered at the generally used NT regions of HC-Pro and CP, and other possibly permissive sites to investigate their effectiveness to express the GFP (jellyfish green fluorescent protein) and Der p 5 (allergen from the dust mite, Dermatophagoides pteronyssinus) ORFs. The results demonstrated the permissiveness of the NT regions of P3, CIP and NIb to carry the ORFs and express the translates as part of the viral polyprotein, the processing of which released free-form proteins in the host cell milieu. However, these sites varied in their permissiveness to retain the ORFs intact and hence affect the heterologous protein expression. Moreover, strong influence of the inserted ORF and host plants in determining the permissiveness of a viral genomic context to stably carry the alien ORFs and hence to support their prolonged expression was also noticed. In general, the engineered sites were relatively more permissive to the GFP ORF than to the Der p 5 ORF. Among the hosts, the local lesion host, Chenopodium quinoa Willd. showed the highest extent of support to TuMV to stably carry the heterologous ORFs at the engineered sites and the protein expression therefrom. Among the systemic hosts, Nicotiana benthamiana Domin proved more supportive to TuMV to carry and express the heterologous ORFs than the Brassica hosts, whereas the protein expression levels were significantly higher and more stable in the plants of Brassica campestris L. var. chinensis and B. campestris L. var. ching-geeng than those in the plants of B. juncea L. and B. campestris L. var. pekinensis.