Proteomic Profiling of Serum from Patients with Tuberculosis

BACKGROUND: Effective treatment and monitoring of tuberculosis (TB) requires biomarkers that can be easily evaluated in blood samples. The aim of this study was to analyze the serum proteome of patients with TB and to identify protein biomarkers for TB. METHODS: Serum samples from 26 TB patients and 31 controls were analyzed by using nano-flow ultra-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry in data-independent mode, and protein and peptide amounts were calculated by using a label-free quantitative approach. The generated data were analyzed by using principal component analysis and partial least squares discriminant analysis, a multivariate statistical method. RESULTS: Of more than 500 proteins identified, alpha-1-antitrypsin was the most discriminative, which was 4.4 times higher in TB patients than in controls. Peptides from alpha-1-antitrypsin and antithrombin III increased in TB patients and showed a high variable importance in the projection scores and coefficient in partial least square discriminant analysis. CONCLUSIONS: Sera from patients with TB had higher alpha-1-antitrypsin levels than sera from control participants. Alpha-1-antitrypsin levels may aid in the diagnosis of TB.

Growth Inhibition of Hepatocellular Carcinoma Huh7 Cells by Lactobacillus casei Extract

PURPOSE: Lactobacillus casei (L. casei) is known to exert anti-proliferation effects on many types of cancer cells. However, the effect of L. casei on liver cancer has not been reported. Accordingly, the aim of this study was to determine the anti-cancer effect of L. casei extract on Huh7 cells. MATERIALS AND METHODS: L. casei ATCC393 extract was prepared and purified. After the treatment of L. casei extract on Huh7 cells, cell viability, cell cycle arrest and cell death were analyzed by flow cytometry. The expression levels of tumor necrosis factor-alpha receptor 1 (TNFR1) and death receptor 3 (DR3) mRNA related with extrinsic apoptosis were assessed by reverse transcription polymerase chain reaction. Additionally, P21 and P27 cell cycle proteins as well as Caspase-3, -8, -9, phospho-Bad and Bcl-2 apoptosis proteins were analyzed by western blot analysis. To determine the effect of L. casei extract on cancer stem-like cells, we analyzed changes in side population fraction through flow cytometry. RESULTS: The cell viability of Huh7 cells treated with L. casei extract was decreased by 77%, potentially owing to increases in the rates of Huh7 cells arrested in the G2/M phase (3% increase) and that underwent apoptosis (6% increase). The expression levels of TNFR1 and DR3 mRNA, as well as P21 and P27 cell cycle proteins, were increased. Meanwhile, the expressions of caspase-8, -9, phospho-Bad and Bcl-2 proteins decreased. However, in the case of side population cells, no remarkable changes were observed. CONCLUSION: L. casei extract exerts a potent anti-tumor effect on the viability of liver cancer cells, although not on cancer stem-like cells.

The Regulation of Cytokine and Chemokine Expressions in Skin Xenograft by Aminoguanidine / 대한이식학회지

PURPOSE: Previous reports demonstrated that nitric oxide (NO) plays immuno-regulatory role in immune responses including allograft rejection response. However, its possible role in xenograft rejection has not been examined. The purpose of this study is to elucidate possible immunoregulatory role of NO in skin xenograft rejection by determining the expressions of chemokines and cytokines in the presence or absence of iNOS inhibitors. METHODS: C57BL/6J mice were grafted with Lewis rat tail skin. The mice were injected intraperitoneally with potent inhibitor of iNOS, aminoguanidine (AMG, 200 mg/kg). Graft survival was monitored and cytokine and chemokine mRNA expressions were measured by real-time RT-PCR in context with iNOS expression on day 3, 5, 7 and 9. These data were compared with those of control mice (saline injected). RESULTS: Compared with the control mice, the AMG treated mice showed delayed xenograft rejection by approximately 3 days (8.9+/-0.7 days vs 11.7+/-1.2 days). Infiltrations of CD11b+, MOMA-2+ cells and neutrophils were significantly reduced but not CD4+ and CD8+ cells in AMG treated graft. The expression of cytokines such as IL-1beta, IL-2, IL-6, IL-12, IFN-gamma in AMG treated graft significantly decreased (P<0.01) whereas IL- 10, TNF-alpha and TGF-beta1 were not changed or enhanced. Additionally, the expression of CC-chemokines such as RANTES and MIP-1alpha significantly reduced (P<0.01) whereas CXC-chemokines such as IP-10 and MIG did not change. CONCLUSION: These data imply that NO suppression by iNOS inhibitor may prolong rat to mouse skin xenograft survival through a selective inhibition of pro-inflammatory cytokines and chemokines. The possible role of NO in transplant rejection can be, therefore, extended to regulation of cytokine and chemokine expressions.

Comparison of the Bacterial Contamination Rates according to the Urine Collection Methods in Women / 기본간호학회지

BACKGROUND: The purpose of this study was to determine whether cleansing the perineum and urethral meatus and using midstream urine affect the rate of bacterial contamination of urine specimens, and to determine the optimum urine collection method. We studied 41 asymptomatic healthy nursing school students. Women who were menstruating were not excluded from this study. METHOD: The first and midstream urinesamples were collected during consecutive urinationsby each woman. The first sample was not a clean-catch specimen, and the second one was a clean-catch specimen. Both specimens were studied by urinalysis and bacterial culture with standard methods. RESULTS: 41 women met the study criteria and 39 successfully completed the study. None of the urine cultures were positive. 68.3% of the non clean-catch first urine cultures, 53.7% of the non clean-catch midstream cultures, 33.3% of the first clean-catch urine culteres and 30.8% of the midstream clean-catch urine were found to be contaminated. There was a significant difference in the bacterial contamination rates between the first and midstream urine, and the clean-catch and non clean-catch urine(p=0.035, p=0.001 respectively). On urinalysis, 7.3% of the non clean-catch first urine, 7.3% of the non clean-catch midstream urine, 2.6% of the clean-catch first urine and 2.6% of clean-catch midstream urine were found to be above grade 2. CONCLUSIONS: According to our results, the bacterial contamination rate was the lowest in midstream and clean catch urine specimens. Threrfore it is recommended that the midstream clean-catch technique is the standard practice for collecting urine specimens for bacterial culture in women.