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Background: DNA methylation has been suggested as a biomarker for early cancer detection and treatment. Varieties of technologies for detecting DNA methylation have been developed, but they are not sufficiently sensitive for use in diagnostic devices. The aim of this study was to determine the suitability of Raman spectroscopy for label-free detection of methylated DNA. Methods: The methylated promoter regions of cancer-related genes cadherin 1 (CDH1) and retinoic acid receptor beta (RARB) served as target DNA sequences. Based on bisulfite conversion, oligonucleotides of methylated or nonmethylated probes and targets were synthesized for the DNA methylation assay. Principal component analysis with linear discriminant analysis (PCA-DA) was used to discriminate the hybridization between probes and targets (methylated probe and methylated target or nonmethylated probe and nonmethylated target) of CDH1 and RARB from nonhybridization between the probe and targets (methylated probe and nonmethylated target or nonmethylated probe and methylated target). Results: This study revealed that the CDH1 and RARB oligo sets and their hybridization data could be classified using PCA-DA. The classification results for CDH1 methylated probe + CDH1 methylated target versus CDH1 methylated probe + CDH1 unmethylated target showed sensitivity, specificity, and error rates of 92%, 100%, and 8%, respectively. The classification results for the RARB methylated probe + RARB methylated target versus RARB methylated probe + RARB unmethylated target showed sensitivity, specificity, and error rates of 92%, 93%, and 11%, respectively. Conclusions: Label-free detection of DNA methylation could be achieved using Raman spectroscopy with discriminant analysis.

Purpose The differentiation properties of stem cells are not yet fully understood due to their close association with multiple environmental and extrinsic factors. This study investigates the differentiation properties of mesenchymal stem cells (MSCs) and correlates them with their intrinsic mechanical properties. Methods A total of 3 different types of MSCs, namely bone marrow-derived MSCs (BMSCs), umbilical cord-derived MSCs (UCSCs), and adipose-derived MSCs (ADSCs) were evaluated. These 3 MSCs were individually differentiated into adipocytes and osteoblasts for 3 weeks. The mechanical properties of the MSCs and differentiated cells were determined by atomic force microscopy. Results ADSCs showed the greatest ability to differentiate into adipocytes, followed by BMSCs and UCSCs. While UCSCs differentiated readily into osteoblasts, BMSCs and ADSCs were less likely to undergo this differentiation. UCSCs were the “hardest” cells, while ADSCs were the “softest.” The cells differentiated from “hard” MSCs were stiffer than the cells differentiated from “soft” MSCs, irrespective of lineage specification. Conclusions The differentiation ability of MSCs and the mechanical properties of the differentiated cells were closely linked. However, there were no significant correlations regarding changes in the mechanical properties between the nuclear region and the cytoplasm during differentiation.

Microfluidics is considered an important technology that is suitable for numerous biomedical applications, including cancer diagnosis, metastasis, drug delivery, and tissue engineering. Although microfluidics is still considered to be a new approach in urological research, several pioneering studies have been reported in recent years. In this paper, we reviewed urological research works using microfluidic devices. Microfluidic devices were used for the detection of prostate and bladder cancer and the characterization of cancer microenvironments. The potential applications of microfluidics in urinary analysis and sperm sorting were demonstrated. The use of microfluidic devices in urology research can provide high-throughput, high-precision, and low-cost analyzing platforms.

Cyanoacrylate (CA) is most widely used as a medical and commercial tissue adhesive because of easier wound closure, good cosmetic results and little discomfort. But, CA-based tissue adhesives have some limitations including the release of cytotoxic chemicals during biodegradation. In previous study, we made prepolymerized allyl 2-CA (PACA) based tissue adhesive, resulting in longer chain structure. In this study, we investigated a biocompatibility of PACA as alternative tissue adhesive for medical application, comparing with that of Dermabond® as commercial tissue adhesive. The biocompatibility of PACA was evaluated for short-term (24 hr) and long-term (3 and 7 days) using conventional cytotoxicity (WST, neutral red, LIVE/DEAD and TUNEL) assays, hematoxylin-eosin (H&E) and Masson trichrome (MT) staining. Besides we examined the biochemical changes in cells and DNA induced by PACA and Dermabond® utilizing Raman spectroscopy which could observe the denaturation and conformational changes in protein, as well as disintegration of the DNA/RNA by cell death. In particular, we analyzed Raman spectrum using the multivariate statistical methods including principal component analysis (PCA) and support vector machine (SVM). As a result, PACA and Dermabond® tissue adhesive treated cells and tissues showed no difference of the cell viability values, histological analysis and Raman spectral intensity. Also, the classification analysis by means of PCA-SVM classifier could not discriminate the difference between the PACA and Dermabond® treated cells and DNA. Therefore we suggest that novel PACA might be useful as potential tissue adhesive with effective biocompatibility.

This study investigated the reversible effects of pulsed radiofrequency (PRF) treatment at 42 °C on the ultrastructural and biological changes in nerve and collagen fibers in the progression of neuropathic pain after rat sciatic nerve injury. Assessments of morphological changes in the extracellular matrices by atomic force microscopy and hematoxylin-eosin, Masson's trichrome and picrosirius-red staining as well as the expressions of two fibril-forming collagens, types-I and -III, and two inflammatory cytokines, TNF-α and IL-6, were evaluated on day 30 after RF exposure. There were four groups for different RF thermal treatments: no treatment, no current, PRF, and continuous RF (CRF). An RF procedure similar to that used in human clinical trials was used in this study. The CRF treatment at 82 °C led to neural and collagen damage by the permanent blockage of sensory nociceptors. The PRF treatment led to excellent performance and high expandability compared to CRF, with effects including slight damage and swelling of myelinated axons, a slightly decreased amount of collagen fibers, swelling of collagen fibril diameters, decreased immunoreactivity of collagen types-I and -III, presence of newly synthesized collagen, and recovery of inflammatory protein immunoreactivity. These evidence-based findings suggest that PRF-based pain relief is responsible for the temporary blockage of nerve signals as well as the preferential destruction of pain-related principal sensory fibers like the Aδ and C fibers. This suggestion can be supported by the interaction between the PRF-induced electromagnetic field and cell membranes; therefore, PRF treatment provides pain relief while allowing retention of some tactile sensation.

The apoptosis of cancer cells is associated with changes in the important cell properties including morphology, surface roughness and stiffness. Therefore, the changes in morphology and biophysical properties can be a good way of evaluating the anticancer activity of a drug. This study examined the effect of paclitaxel on the properties of Ishikawa and HeLa cells using atomic force microscopy (AFM), and the relationship between the changes in morphology and the biophysical properties and apoptosis was discussed. The viability and proliferation of the cells were analyzed using the methylthiazol tetrazolium (MTT) method and a TUNEL assay to confirm cellular apoptosis due to a paclitaxel treatment. AFM observations clearly showed the apoptotic morphological and biophysical changes in Ishikawa and HeLa cells. After the paclitaxel treatment, the cell membrane was torn and holed, the surface roughness was increased, and the stiffness was decreased. These changes were observed more apparently after a 24 h treatment and in Ishikawa cells compared to HeLa cells. The MTT and TUNEL assays results revealed the Ishikawa cells to be more sensitive to paclitaxel than HeLa cells and definite apoptosis occurred after a 24 h treatment. These results showed good agreement with the AFM results. Therefore, research on the morphological and biophysical changes by AFM in cancer cells will help to evaluate the anticancer activities of the drugs.

The antifungal activity of allicin and its synergistic effects with the antifungal agents flucytosine and amphotericin B (AmB) were investigated in Candida albicans (C. albicans). C. albicans was treated with different conditions of compounds alone and in combination (allicin, AmB, flucytosine, allicin + AmB, allicin + flucytosine, allicin + AmB + flucytosine). After a 24-hour treatment, cells were examined by scanning electron microscopy (SEM) and atomic force microscopy (AFM) to measure morphological and biophysical properties associated with cell death. The clearing assay was conducted to confirm the effects of allicin. The viability of C. albicans treated by allicin alone or with one antifungal drug (AmB, flucytosine) in addition was more than 40% after a 24-hr treatment, but the viability of groups treated with combinations of more than two drugs was less than 32%. When the cells were treated with allicin alone or one type of drug, the morphology of the cells did not change noticeably, but when cells were treated with combinations of drugs, there were noticeable morphological changes. In particular, cells treated with allicin + AmB had significant membrane damage (burst or collapsed membranes). Classification of cells according to their cell death phase (CDP) allowed us to determine the relationship between cell viability and treatment conditions in detail. The adhesive force was decreased by the treatment in all groups compare to the control. Cells treated with AmB + allicin had a greater adhesive force than cells treated with AmB alone because of the secretion of molecules due to collapsed membranes. All cells treated with allicin or drugs were softer than the control cells. These results suggest that allicin can reduce MIC of AmB while keeping the same efficacy.

The death process of Candida albicans was investigated after treatment with the antifungal agents flucytosine and amphotericin B by assessing morphological and biophysical properties associated with cell death. C. albicans was treated varying time periods (from 6 to 48 hours) and examined by scanning electron microscopy (SEM) and atomic force microscopy (AFM). SEM and AFM images clearly showed changes in morphology and biophysical properties. After drug treatment, the membrane of C. albicans was perforated, deformed, and shrunken. Compared to the control, C. albicans treated with flucytosine was softer and initially showed a greater adhesive force. Conversely, C. albicans treated with amphotericin B was harder and had a lower adhesive force. In both cases, the surface roughness increased as the treatment time increased. The relationships between morphological changes and the drugs were observed by AFM clearly; the surface of C. albicans treated with flucytosine underwent membrane collapse, expansion of holes, and shrinkage, while the membranes of cells treated with amphotericin B peeled off. According to these observations, the death process of C. albicans was divided into 4 phases, CDP(0), CDP(1), CDP(2), and CDP(4), which were determined based on morphological changes. Our results could be employed to further investigate the antifungal activity of compounds derived from natural sources.

Voltage-dependent anion channel (VDAC) is the main protein in mitochondria-mediated apoptosis, and the modulation of VDAC may be induced by the excessive release of extracellular glutamate. This study examined the role of glutamate release on VDAC-mediated apoptosis in an eleven vessel occlusion model in rats. Male Sprague-Dawley rats (250-350 g) were used for the 11 vessel occlusion ischemic model, which were induced for a 10-min transient occlusion. During the ischemic and initial reperfusion episode, the real-time monitoring of the extracellular glutamate concentration was measured using an amperometric microdialysis biosensor and the cerebral blood flow (CBF) was monitored by laser-Doppler flowmetry. To confirm neuronal apoptosis, the brains were removed 72 h after ischemia to detect the neuron-specific nuclear protein and pro-apoptotic proteins (cleaved caspase-3, VDAC, p53 and BAX). The changes in the mitochondrial morphology were measured by atomic force microscopy. A decrease in the % of CBF was observed, and an increase in glutamate release was detected after the onset of ischemia, which continued to increase during the ischemic period. A significantly higher level of glutamate release was observed in the ischemia group. The increased glutamate levels in the ischemia group resulted in the activation of VDAC and pro-apoptotic proteins in the hippocampus with morphological alterations to the mitochondria. This study suggests that an increase in glutamate release promotes VDAC-mediated apoptosis in an 11 vessel occlusion ischemic model.

We present a method for improving the lithographic resolution of digital micromirror devices for flow-lithography using a wobulation technique. While maintaining the area of UV exposure, the lithographic resolution was improved using a wobulation technique, which is a large screen display technique that enhances resolution via overlapping pixels by half a pixel. The edges of a diagonal pattern in a microstructure were smoothly generated with additional sub-patterns compared to conventional single pattern-exposure. In addition, the surface roughness of the microstructure was improved because the gaps between pixels were filled by the overlapping patterns. [ABSTRACT FROM AUTHOR]

Atmospheric pressure non-thermal plasma has been introduced in various applications such as wound healing, sterilization of infected tissues, blood coagulation, delicate surgeries, and so on. The nonthermal plasma generates reactive oxygen species (ROS), including ozone. Various groups have reported that the produced ROS influence proliferation and differentiation of cells, as well as apoptosis and growth arrest of tumor cells. In this study, we investigated the effects of non-thermal plasma on rheological characteristics of red blood cells (RBC). We experimentally measured the extent of hemolysis, deformability, and aggregation of red blood cells (RBC) with respect to exposure times of non-thermal plasma. RBC morphology was also examined using field-emission scanning electron microscopy. The absorbance of hemoglobin released from the RBCs increased with increasing exposure time of the non-thermal plasma. Values of the elongation index and aggregation index were shown to decrease significantly with increasing plasma exposure times. Therefore, hemorheological properties of RBCs could be utilized to assess the performance of various non-thermal plasmas. [ABSTRACT FROM AUTHOR]

Human adipose-derived stem cells (hASCs) have a capacity to undergo adipogenic, chondrogenic, and osteogenic differentiation. Recently, hASCs were applied to various fields including cell therapy for tissue regeneration. However, it is hard to predict the direction of differentiation of hASCs in real-time. Matrix metalloproteinases (MMPs) are one family of proteolytic enzymes that plays a pivotal role in regulating the biology of stem cells. MMPs secreted by hASCs are expected to show different expression patterns depending on the differentiation state of hASCs because biological functions exhibit different patterns during the differentiation of stem cells. Here, we investigated proteolytic enzyme activity, especially MMP-2 activity, in hASCs during their differentiation. The activities of proteolytic enzymes and MMP-2 were higher during chondrogenic differentiation than during adipogenic and osteogenic differentiation. During chondrogenic differentiation, mRNA expression of MMP-2 and the level of the active form of MMP-2 were increased, which also correlated with Col II. It is concluded that proteolytic enzyme activity and the level of the active form of MMP-2 were increased during chondrogenic differentiation, which was accelerated in the presence of Col II protein. According to our findings, MMP-2 could be a candidate maker for real-time detection of chondrogenic differentiation of hASCs. [ABSTRACT FROM AUTHOR]

The extracellular matrix (ECM) of cartilage performs essential functions in differentiation and chondroprogenitor cell maintenance during development and regeneration. Here, we discuss the vital role of matrilin-3, an ECM protein involved in cartilage development and potential osteoarthritis pathomechanisms. As an adaptor protein, matrilin-3 binds to collagen IX to form a filamentous network around cells. Matrilin-3 is an essential component during cartilage development and ossification. In addition, it interacts directly or indirectly with transforming growth factor β (TGF-β), and bone morphogenetic protein 2 (BMP2) eventually regulates chondrocyte proliferation and hypertrophic differentiation. Interestingly, matrilin-3 increases interleukin receptor antagonists (IL-Ra) in chondrocytes, suggesting its role in the suppression of IL-1β-mediated inflammatory action. Matrilin-3 downregulates the expression of matrix-degrading enzymes, such as a disintegrin metalloproteinase with thrombospondin motifs 4 (ADAMTS4) and ADAMTS5, matrix metalloproteinase 13 (MMP13), and collagen X, a hypertrophy marker during development and inflammatory conditions. Matrilin-3 essentially enhances collagen II and aggrecan expression, which are required to maintain the tensile strength and elasticity of cartilage, respectively. Interestingly, despite these attributes, matrilin-3 induces osteoarthritis-associated markers in chondrocytes in a concentration-dependent manner. Existing data provide insights into the critical role of matrilin-3 in inflammation, matrix degradation, and matrix formation in cartilage development and osteoarthritis. [ABSTRACT FROM AUTHOR]

In this study, we developed poly(ε-caprolactone) (PCL) 3D scaffolds using a solid free form fabrication (SFF) technique. β-cyclodextrin (βCD) was grafted to hydroxyapatite (HAp) and this βCD grafted HAp was coated onto the PCL scaffold surface, followed by drug loading through an inclusion complex interaction between the βCD and adamantane (AD) or between βCD and simvastatin (SIM). The scaffold structure was characterized by scanning electron microscopy (SEM). The release profile of simvastatin in the β-CD grafted HAp was also evaluated. Osteogenic differentiation of adipose-derived stromal cells (ADSCs) was examined using an alkaline phosphatase activity (ALP) assay. The results suggest that drug loaded PCL-HAp 3-D scaffolds enhances osteogenic differentiation of ADSCs. [ABSTRACT FROM AUTHOR]

Polymers have been used extensively taking forms as scaffolds, patterned surface and nanoparticle for regenerative medicine applications. Angiogenesis is an essential process for successful tissue regeneration, and endothelial cell-cell interaction plays a pivotal role in regulating their tight junction formation, a hallmark of angiogenesis. Though continuous progress has been made, strategies to promote angiogenesis still rely on small molecule delivery or nuanced scaffold fabrication. As such, the recent paradigm shift from top-down to bottom-up approaches in tissue engineering necessitates development of polymer-based modular engineering tools to control angiogenesis. Here, we developed cationic nanocylinders (NCs) as inducers of cell-cell interaction and investigated their effect on angiogenic activities of human umbilical vein endothelial cells (HUVECs) in vitro. Electrospun poly (L-lactic acid) (PLLA) fibers were aminolyzed to generate positively charged NCs. The aninolyzation time was changed to produce two different aspect ratios of NCs. When HUVECs were treated with NCs, the electrostatic interaction of cationic NCs with negatively charged plasma membranes promoted migration, permeability and tubulogenesis of HUVECs compared to no treatment. This effect was more profound when the higher aspect ratio NC was used. The results indicate these NCs can be used as a new tool for the bottom-up approach to promote angiogenesis. [ABSTRACT FROM AUTHOR]

Estrogens and estrogen receptors (ESRs) have been implicated in the stimulation of aberrant prostate growth and the development of prostate diseases. The aim of the present study was to investigate four single nucleotide polymorphisms (SNPs) of the ESR2 gene in order to examine whether ESR2 is a susceptibility gene for benign prostatic hyperplasia (BPH). In order to evaluate whether an association exists between ESR2 and BPH risk, four polymorphisms [rs4986938 (intron), rs17766755 (intron), rs12435857 (intron) and rs1256049 (Val328Val)] of the ESR2 gene were genotyped by direct sequencing. A total of 94 patients with BPH and 79 control subjects were examined. SNPStats and Haploview version 4.2 we used for the genetic analysis. Multiple logistic regression models (codominant1, codominant2, dominant, recessive and log-additive) were produced in order to obtain the odds ratio, 95% confidence interval and P-value. Three SNPs (rs4986938, rs17766755 and rs12435857) showed significant associations with BPH (rs4986938, P=0.015 in log-additive model; rs17766755, P=0.033 in codominant1 model, P=0.019 in dominant model and P=0.020 in log-additive model; rs12435857, P=0.023 in dominant model and P=0.011 in log-additive model). The minor alleles of these SNPs increased the risk of BPH, and the AAC haplotype showed significant association with BPH (?2=6.34, P=0.0118). These data suggest that the ESR2 gene may be associated with susceptibility to BPH. [ABSTRACT FROM AUTHOR]

The study was to investigate the changes in biochemical properties of activated mature CD8+ T cells related to apoptosis at a molecular level. We confirmed the activation and apoptosis of CD8+ T cells by fluorescence- activated cell sorting and atomic force microscopy and then performed Raman spectral measurements on activated mature CD8+ T cells and cellular deoxyribose nucleic acid (DNA). In the activated mature CD8+ T cells, there were increases in protein spectra at 1002 and 1234 cm-1. In particular, to assess the apoptosisrelated DNA spectral signatures, we investigated the spectra of the cellular DNA isolated from resting and activated mature CD8+ T cells. Raman spectra at 765 to 786 cm-1 and 1053 to 1087 cm-1 were decreased in activated mature DNA. In addition, we analyzed Raman spectrum using the multivariate statistical method including principal component analysis. Raman spectra of activated mature DNA are especially well-discriminated from those of resting DNA. Our findings regarding the biochemical and structural changes associated with apoptosis in activated mature T cells and cellular DNA according to Raman spectroscopy provide important insights into allospecific immune responses generated after organ transplantation, and may be useful for therapeutic manipulation of the immune response. [ABSTRACT FROM AUTHOR]

We demonstrate the fabrication of an all-carbon electrode by plasma-enhanced chemical vapor deposition for use in flexible electrochemical applications. The electrode is composed of vertically aligned carbon nanotubes that are grown directly on a flexible graphite foil. Being all-carbon, the simple fabrication process and the excellent electrochemical characteristics present an approach through which high-performance, highly-stable and cost-effective electrochemical applications can be achieved. [ABSTRACT FROM AUTHOR]

The exact mechanism of rheumatoid arthritis (RA) is unclear, but a combination of genetic, environmental and hormonal factors is thought to be involved. This study examined the progressive arthritic reaction of murine type II collagen-induced arthritis (CIA), a representative animal model of RA. Arthritic reactions, including inflammation and bone erosion were examined using an objective non-invasive method. Two scoring systems were used to evaluate changes in cutaneous inflammation and bone erosion during RA progression. The severity of inflammation was evaluated by visual scoring of erythema and edema, while the degree of bone erosion was quantified by macroradiographical erosion analysis of specific bones. A significant difference was observed in both visual ( P = 0.0001, n = 7) and radiographic ( P < 0.0001, n = 7) examinations for the RA group as compared to the control. The relationship between inflammatory change and erosive change in bone showed a significant positive correlation, r = 0.9550 ( P < 0.0001, n = 7). The overall rate of asymmetry was 25.23% in both fore- and hindpaws. The results generated from these experiments show that murine CIA is a promising model for elucidating the mechanism of RA. In addition, the results of this study may be used for monitoring RA progression as well as screening therapy efficacy in the joint pathology. [ABSTRACT FROM AUTHOR]

Objective: This study presents a method for non-invasive screening of progressive arthritic reactions in a representative rheumatoid arthritis (RA) animal model of Type II collagen-induced arthritis (CIA). Materials: Seventeen male DBA/1J mice (9 weeks old) were used for the control group (seven) and the CIA group (ten). Treatment: The CIA model was induced by the injection of a Type II collagen and incomplete Freund's adjuvant emulsion. Methods: Arthritic reactions including joint space narrowing (JSN), bone erosion, inflammation, and physiological defects were observed using five non-invasive methods. The degrees of JSN and bone erosion were quantified using macro-radiographs of specific joints and bones, while the severity of inflammation was evaluated by changes in paw volume and visual scoring of edema and erythema. Results: Significant differences were observed in JSN (≤48.29%, p < 0.0001), bone erosion (78.30%, p < 0.0001), grip strength ( p < 0.05), and inflammation (91.67% in edema and 194.12% increase in paw volume, p < 0.001) between the CIA group and the controls. The relationships between factors exhibited significant positive correlations ( R ≥ 0.9229, p < 0.05) between radiographic parameters, with symmetries of ≥0.9062 ( p < 0.05) for JSN and ≥ 0.8575 ( p < 0.001) for bone erosion, 0.8770 ( p < 0.0001) for inflammatory parameters, and ≥0.9527 ( p < 0.0001) for radiographic and non-radiographic parameters. Conclusions: Our non-invasive radiographic and non-radiographic methods of documentation were both acceptable for screening joint defects during RA progression. In addition, the results of this study may be useful for monitoring the progression of joint pathology in RA diseases. [ABSTRACT FROM AUTHOR]

Background/Aims: In humans, the kidneys regulate blood pressure by balancing sodium concentrations. Fine-tuning of renal sodium reabsorption and excretion depends on the epithelial sodium channel protein (ENaC: protein complex of SCNN1A, SCNN1B, and SCNN1G). The surface expression of ENaC components is directed by the ubiquitination of ENaC by NEDD4L, an ENaC-specific E3 ubiquitin ligase, and is regulated by the deubiquitination of ENaC by USP2. The activity of NEDD4L in turn is regulated by phosphorylation by SGK1 and also through interaction with NDFIP2. Methods: We analyzed 91 SNPs in 7 genes using the genotype data of 8,842 individuals from the Korea Association REsource subject pool for their correlation with blood pressure and hypertension. Results: 25 SNPs in the SCNN1A, SCNN1B, SCNN1G, NEDD4L, NDFIP2, and USP2 loci were found to be associated with blood pressure. An additional hypertension case-control study identified 13 SNPs in SCNN1B, SCNN1G, and NEDD4L that were linked to hypertension. Conclusion: These results support our hypothesis that individual variations in blood pressure are attributed to variants of the genes that regulate renal sodium reabsorption and excretion. Our data also suggest that it would be meaningful to investigate the role of NEDD4L-mediated ubiquitination in the pathogenesis of hypertension. Copyright © 2010 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Background: The prevalence of type 2 diabetes has reached epidemic proportions worldwide, and the incidence of life-threatening complications of diabetes through continued exposure of tissues to high glucose levels is increasing. Advances in genotyping technology have increased the scale and accuracy of the genotype data so that an association genetic study has expanded enormously. Consequently, it is difficult to search the published association data efficiently, and several databases on the association results have been constructed, but these databases have their limitations to researchers: some providing only genome-wide association data, some not focused on the association but more on the integrative data, and some are not user-friendly. In this study, a userfriend database of type 2 diabetes genetic association of manually curated information was constructed. Description: The list of publications used in this study was collected from the HuGE Navigator, which is an online database of published genome epidemiology literature. Because type 2 diabetes genetic association database (T2DGADB) aims to provide specialized information on the genetic risk factors involved in the development of type 2 diabetes, 701 of the 1,771 publications in the type 2 Diabetes case-control study for the development of the disease were extracted. Conclusions: In the database, the association results were grouped as either positive or negative. The gene and SNP names were replaced with gene symbols and rsSNP numbers, the association p-values were determined manually, and the results are displayed by graphs and tables. In addition, the study design in publications, such as the population type and size are described. This database can be used for research purposes, such as an association and functional study of type 2 diabetes related genes, and as a primary genetic resource to construct a diabetes risk test in the preparation of personalized medicine in the future. [ABSTRACT FROM AUTHOR]

Melatonin possesses a number of important biologic activities including oncostatic, anti-oxidant, and immunostimulatory actions. This study was designed to assess the effects of melatonin on inflammation-related gene expression in lipopolysaccharide (LPS)-stimulated human peripheral blood mononuclear cells (PBMCs), using CombiMatrix 2K Human Inflammation chip. After pretreatment with melatonin (100 μm) for 4 hr, cells were incubated with LPS (1 μg/mL) for 24 hr. We compared gene expression profiles between LPS-treated, melatonin-treated, LSP/melatonin-treated, and control groups. LPS induced the upregulation of 95 genes, compared with controls. Melatonin pretreatment in LPS-stimulated PBMCs suppressed the expression of 23 genes more than twofold. Interestingly, melatonin showed a suppressive effect on the expression of CC chemokine subfamily genes, including CCL2/MCP1, CCL3/MIP1 α, CCL4/MIP1 β, CCL5/RANTES, CCL8/MCP2, CCL20/MDC, and CCL22/MIP3 α, in LPS-stimulated PBMCs. This result was confirmed by reverse transcriptase polymerase chain reaction. Among the CC chemokine subfamily genes, particularly, the expression of CCL2 and CCL5 was markedly downregulated by melatonin in LPS-stimulated PBMCs. The secretion levels of CCL2 and CCL5 were measured using enzyme-linked immunosorbent assay. Stimulation of PBMCs by LPS induced the secretion of CCL2 (2334.3 ± 161.4 pg/mL, mean ± S.E.M.), whereas melatonin pretreatment (153.0 ± 3.8 pg/mL) inhibited the LPS-induced secretion of CCL2. Melatonin pretreatment (2696.2 ± 385.3 pg/mL) also inhibited the LPS-induced secretion of CCL5 (4679.6 ± 107.5 pg/mL). Taken together, these results suggest that melatonin may have a suppressive effect on LPS-induced expression of CC chemokine genes, especially CCL2 and CCL5, which may explain its beneficial effects in the treatment of various inflammatory conditions. [ABSTRACT FROM AUTHOR]

Melatonin is reported to be an anti-inflammatory agent. No genetic study concerning the association between melatonin and inflammatory disease has yet been reported. Here we performed a polymorphism study on the melatonin receptor type 1B ( MTNR1B) in Korean rheumatoid arthritis (RA) patients and controls. The polymorphism of MTNR1B located in 3′-untranslated region (rs 1562444) was selected for its higher rate of heterozygosity among other single nucleotide polymorphisms (SNPs) in both MTNR1A and MTNR1B genes and investigated in RA patients (n = 173) and healthy controls (n = 195) by polymerase chain reaction-restriction fragment length polymorphism assay using NlaIII restriction enzyme. No statistically significant difference in either genotype distribution or allele frequency was observed between RA patients and controls. The genotype distributions and allele frequencies of rheumatoid factor negative [RF(−)] patients were similar to those of controls. However, statistical analysis of genotype revealed a significant association ( χ2 = 6.42, P = 0.04) is present between RF(+) and MTNR1B SNP (rs 1562444). Although no statistically significant difference in allele frequency between RF(+) and controls was observed ( χ2 = 2.75, P = 0.10), the results might suggest that MTNR1B SNP (rs 1562444) is associated with the presence of RF in RA. This is the first study, to our knowledge, to report a positive genetic relationship between melatonin and RA. [ABSTRACT FROM AUTHOR]

Vitiligo (leukoderma) is an acquired idiopathic hypomelanotic disorder characterized by the circumscribed depigmented patches. Vitiligo is a polygenic disease. The exact pathogenesis is not yet known. The angiotensin converting enzyme (ACE) gene was selected as a candidate gene as ACE plays an important role in the physiology of the vasculature, blood pressure and inflammation, and its relationship with various diseases, including autoimmune diseases, has been widely investigated. The I/D polymorphism of ACE gene in vitiligo patients has not been reported. In this study, we investigated ACE gene polymorphism in 120 vitiligo patients and in 429 healthy volunteers in Korea. The ACE gene genotype distribution (P = 0.032) and allele frequency (P = 0.012) were significantly different between vitiligo patients and healthy controls. This study suggests that the ACE gene polymorphism has a strong association with the development of vitiligo in Korean patients. [ABSTRACT FROM AUTHOR]

There are several applications for non-thermal plasma at atmospheric pressure, such as the sterilization of infected tissues, wound healing, and delicate surgery. Non-thermal plasma generates reactive oxygen/nitrogen species (RONS). In this study, we examined human red blood cells (RBCs), platelet function, and blood coagulation, following treatments with different feeding gases (air, nitrogen (N2), and argon (Ar)) of the dielectric barrier discharge plasma (DBDP) at different exposure times. This is the first study to evaluate the effects of DBDP on hemolysis, deformability, aggregation, and morphology of RBCs, as well as platelet function and blood coagulation at three doses (5, 10 and 15 min exposure). In comparison to N2 and Ar DBDP, air DBDP at more than 10 min exposure to RBC in culture plate with 2 ml liquid solution caused hemolysis and alterations in RBC deformability and aggregation owing to the higher levels of hydrogen peroxide, hydroxyl oxidants and other RONS. Moreover, air DBDP affected platelet function at high doses (more than 10 min exposure). Consequently, the air DBDP at, or more, than 10 min exposure affects blood coagulation and blood cells. The proposed method is useful for assessing the performances and dose optimization of various non-thermal plasma types used with different type of gases in biological applications. [ABSTRACT FROM AUTHOR]

A chart and a diagram are presented that depict a schematic model of carboxyl group, polycaprolactone and cyclodextrin.

Although microfluidic systems have been important tools in analytical chemistry, life sciences, and medical research, their application was rather limited for drug-screening and biosensors. Here, we described a microfluidic device consisting of a multilayer micro-channel system that represented the hemodynamic cerebral vascular system. We analyzed wall shear stresses related to aneurysm formation in the circle of Willis (CoW) and their morphological variations using this system. This device was controlled by pneumatic valves, which occluded various major arteries by closing the associated channels. The hemodynamic analysis indicated that higher degrees of shear stress occurred in an anterior communicating artery (ACoA), particularly in the hypoplastic region of the posterior communicating artery (PCoA) and the P1 segment. Furthermore, occlusion of a common carotid artery (CCA) or a middle cerebral artery (MCA) increased the shear stress, whereas occlusion of a vertebral artery (VA) decreased the shear stress. These results indicate that the morphological variation of the CoW may affect aneurysm formation resulting from increased wall shear stress. Therefore, the technique described in this paper provides a novel method to investigate the hemodynamics of complex cerebral vascular systems not accessible from previous clinical studies. [ABSTRACT FROM AUTHOR]

Prostaglandin E1 (Alprostadil®, PGE1) is often used for patients with critical limb ischemia due to its vasodilator and anti-platelet effects. Although PGE1 is considered to have a cardioprotective effect against myocardial ischemia-reperfusion (IR) injury, its mechanism and correlation with nitric oxide (NO) for cardioprotection are not clear. In this study, we used sol-gel-modified electrochemical NO and O2 microsensors to simultaneously measure the changes in NO level and oxygen tension (pO2) in the myocardium during myocardial IR that were induced by PGE1 pretreatment. Rat hearts were randomly divided into two groups: a control group (n = 5) and a PGE1 group (n = 5, with PGE1 pretreatment). Myocardium that underwent PGE1 pretreatment (26.0 ± 13.1%, p < 0.001) demonstrated more highly reduced endogenous NO production during the ischemic period than that of the control group (86.7 ± 18.5%). After the onset of reperfusion, %NO level in the PGE1 group increased to a maximum of 82.0 ± 6.4 but did not exceed the pre-ischemic basal NO level. However, in the control group, the maximum %NO response (164.9 ± 41.0) on reperfusion was approximately double that of the PGE1 group (p < 0.01, n = 5). The restoration value of pO2 in the PGE1 group significantly increased and was restored to pre-ischemic level (82.2 ± 21.3%); however, the pO2 of the control group did not increase throughout the reperfusion period (12.3 ± 7.9%, p < 0.001). As a result, we suggest that the cardioprotective effect of PGE1 might be attributed to a reduction in excessive NO production during reperfusion. Therefore, simultaneous and real-time monitoring of NO and pO2 dynamics in the myocardium would be effective in evaluating the effects of therapeutic treatments such as drug administration. Also, the reduction in excessive NO release during the early reperfusion period might be helpful as a therapeutic strategy to protect the myocardium from IR injury. [ABSTRACT FROM AUTHOR]

Because the cell membrane is an important regulator of cell function, its morphological changes are important markers of cell apoptosis. These changes can differ for each cell type, and depend on the treatment conditions, including the drug, doses, and treatment time. To quantify morphological changes, HeLa and Ishikawa cells were investigated with atomic force microscopy. Both cells were treated with cisplatin (1 mM) for 24 hours. The viability and proliferation of the cells were analyzed with methylthiazol tetrazolium method. The proliferation rates of both cells treated with cisplatin decreased more than 50%. The morphological changes induced by cisplatin were dependent on the cell type, and the results were determined quantitatively. The surface of HeLa cells became rougher with cisplatin treatment, whereas cisplatin-treated Ishikawa cells were smoother than untreated cells. In both cases, cell height was decreased with cisplatin treatment. These results suggest that atomic force microscopy can be used to analyze anticancer drug activity in cancer cells. (J Nippon Med Sch 2012; 79: 320?326) [ABSTRACT FROM AUTHOR]