Biomarkers and Genomic Medicine最新文献

The prevalence of diabetes around the world is increasing, and the complications of diabetes are becoming worse so that the global burden of diabetes-related complications is rising. The purpose of this study was to simulate the physiological condition of diabetes in the human body by culturing human microvascular endothelial cell 1 (HMEC-1) cells in Dulbecco’s modified Eagle’s medium (DMEM) containing 5.5mM glucose, 25mM glucose, and 50mM glucose. The cell viability of HMEC-1 cells at the indicated glucose concentrations was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The specific epithelial cell junction N-cadherin was measured by immunofluorescence. Furthermore, monocytic initiation of inflammatory reactions was studied by using Western blotting and enzyme-linked immunosorbent assay. Based on our results, hyperglycemia treatment influenced the distribution of N-cadherin-containing structures in HMEC-1 cells, whereas the change of the inflammatory profiles in HMEC-1 cells was affected after their coculture with the supernatant of diabetic THP-1 monocytes. In this study, we conclude that the removal of endothelial N-cadherin caused by hyperglycemia (at the 25mM glucose and 50mM glucose concentrations) may lead to endothelial dysfunction and subsequently endothelial cell death in the late stage of diabetes. Furthermore, the loss of intercellular adhesion molecule 1 and the upregulation of interleukin (IL)-1, IL-6, and tumor necrosis factor alpha mediated by diabetic monocytes showed altered interaction between the vascular endothelium and blood cells in the diabetic microenvironment.

The Charnoly body (CB) is a pleomorphic, electron-dense, multilamellar, preapoptotic, mitochondrial biomarker of cell injury. Nutritional stress and environmental toxins induce CB formation in highly vulnerable developing neurons because of compromised mitochondrial bioenergetics; however, nutritional rehabilitation, physiological zinc supplementation, and metallothioneins (MTs) inhibit CB formation. Accumulation of CBs at the junction of the axon hillock may impair the axoplasmic transport of ions, neurotransmitters, neurotropic factors, and enzymes at the synaptic terminals. Therefore, drugs may be developed to inhibit CB formation in neurodegenerative and cardiovascular diseases. In addition, nonspecific induction of CB formation in hyperproliferating cells with cancer chemotherapy causes as adverse effects alopecia, myelosuppression, gastrointestinal tract symptoms, cardiovascular toxicity, and infertility. Hence, drugs may be developed to induce cancer stem cell-specific CB formation to cure multidrug-resistant malignancies and chronic infections. Natural abundance and genetic susceptibility of mitochondrial DNA qualify CB as an early, unique, and sensitive universal biomarker of clinical significance.

This study was undertaken to assess the frequency of angiotensin converting enzyme (ACE) insertion/deletion (I/D) polymorphism in patients with type 2 diabetes mellitus. A total of 162 patients with type 2 diabetes and 160 individuals without this disease were analyzed. From the diabetes group, 81 patients with cardiovascular risk (according to American Diabetes Association parameters) were selected to form another subgroup. For polymorphism identification, two polymerase chain reactions were performed: one reaction to identify all genotypes and a second one to confirm the presence of the I allele. The observed genotype frequencies were as follows: diabetes group I/I (19.1%), I/D (52.5%), D/D (28.4%); control group I/I (12.5%), I/D (55.6%), D/D (31.9%); and diabetes with cardiovascular risk group I/I (16.0%), I/D (59.3%), D/D (24.7%). No statistically significant difference was observed between the allelic and genotypic frequencies in the analyzed groups. Previous studies reported an association between the D allele and type 2 diabetes in Caucasian and East Asian populations. However, in mixed populations, such as those found in Brazil, such an association was not found. This fact does not discard the need for more studies on the frequencies of this polymorphism in the Brazilian population and the associations with risk factors, which can compromise the quality of life of diabetes patients.

The purpose of this study is to determine the role of curcumin in the alteration of levels of interleukin (IL)-1α, IL-6, and tumor necrosis factor α (TNF-α) as proinflammatory cytokines in monocyte culture exposed to preeclamptic plasma, as well as the effect on the transcription factors: nuclear factor kappa beta (NF-κB) and peroxisome proliferator-activated receptor γ (PPAR-γ). Plasma was taken from preeclamptic women (n = 12) and normotensive pregnant women (n = 12). Monocyte cultures were taken from nonpregnant healthy woman. Monocyte cultures were incubated with plasma for 48 hours. Curcumin in various doses were given in monocytic cultures prior to and after preeclamptic plasma exposure. The levels of IL-1α, IL-6, and TNF-α as well as NF-κB and PPAR-γ in each culture were determined by enzyme-linked immunosorbent assay procedures. The final data were analyzed by analysis of variance (ANOVA) and path analysis. This study shows a significant increase (p < 0.05) in the levels of proinflammatory cytokines (IL-1α, IL-6, and TNFα) in monocyte cultures exposed to preeclamptic plasma compared with normotensive pregnancy plasma. Curcumin treatment in various doses can decrease significantly (p < 0.05) proinflammatory cytokine levels in monocyte cultures that have been already stimulated by preeclamptic plasma. After curcumin treatment, there was a decreased level of nuclear NF-κB p50 and a significantly increased level of PPAR-γ. Curcumin has a direct effect on decreasing the levels of nuclear NF-κB p50 and also curcumin indirectly influenced the level of nuclear NF-κB p50 by the increased level of PPAR-γ. Curcumin could decrease levels of proinflammatory cytokines (IL-1α, IL-6, and TNFα) in monocyte cultures exposed to preeclamptic plasma by affecting the transcription factors, NF-κB and PPAR-γ. Curcumin has potential in the prevention and future treatment of preeclampsia, through inflammation pathways assumed as being responsible for the development of preeclampsia.

In this study, we designed a novel colorimetric method to detect multidrug resistance in Mycobacterium tuberculosis isolates. The assay of loop-mediated isothermal amplification (LAMP) is used to amplify target DNA from multidrug-resistant M. tuberculosis isolates, and enzyme-linked immunosorbent assay (ELISA) is used for the colorimetric determination. This method is designed based on point mutation at the hot spot region in target drug-resistant gene using LAMP-polymerase chain reaction (PCR), hybridization, and thermal melting for differentiating homoduplex DNA (drug-susceptible stain) and heteroduplex DNA (resistance mutant). From ELISA colorimetric detection, color change developed in drug-susceptible strains, and colorless result appeared in resistance mutants. A comparison of this LAMP-PCR-hybridization–thermal melt–ELISA (LAMP–TM–ELISA) method with the automated BACTEC MGIT 960 system showed that the sensitivity of this molecular analysis of resistance to isoniazid, rifampin, amikacin, and ciprofloxacin in M. tuberculosis was 92.3%, 95.3%, 93.1%, and 91.4%, respectively. This method for detection of resistance to isoniazid, rifampin, amikacin, and ciprofloxacin in M. tuberculosis showed a specificity of 95.5–98.2% and a test efficiency of 93.2–96.8%. This LAMP–TM–ELISA method will be a useful tool for rapid diagnosis (within 1 working day) and cost-effectiveness (US$15/reaction) to detect resistance to isoniazid, rifampin, amikacin, and ciprofloxacin via katG, inhA and mabA-inhA promoter, rpoB, rrs, gyrA, and gyrB genes in M. tuberculosis isolates.