Toxic detection in mine water based on proteomic analysis of lysosomal enzymes in Saccharomyces cerevisiae

OBJECTIVES: Lysosome is the cell-organelle which is commonly used as biomonitoring tool in environmental pollution. In this study, the lysosomal proteomic of the yeast Saccharomyces cerevisiae was analyzed for utilization in the detection of toxic substances in mine water samples. METHODS: This work informs the expression of lysosomal proteomic in yeast in response with toxic chemicals, such as sodium meta-arsenite and tetracycline, for screening specific biomarkers. After that, a recombinant yeast contained this biomarker were constructed for toxic detection in pure toxic chemicals and mine water samples. RESULTS: Each chemical had an optimal dose at which the fluorescent protein intensity reached the peak. In the case of water samples, the yeast showed the response with sample 1, 3, 4, and 5; whereas there is no response with sample 2, 6, and 7. CONCLUSIONS: The recombinant yeast showed a high ability of toxic detection in response with several chemicals such as heavy metals and pharmaceuticals. In the case of mine water samples, the response varied depending on the sample content.

Imaging Mass Spectrometry in Papillary Thyroid Carcinoma for the Identification and Validation of Biomarker Proteins

Direct tissue imaging mass spectrometry (IMS) by matrix-assisted laser desorption ionization and time-of-flight (MALDI-TOF) mass spectrometry has become increasingly important in biology and medicine, because this technology can detect the relative abundance and spatial distribution of interesting proteins in tissues. Five thyroid cancer samples, along with normal tissue, were sliced and transferred onto conductive glass slides. After laser scanning by MALDI-TOF equipped with a smart beam laser, images were created for individual masses and proteins were classified at 200-microm spatial resolution. Based on the spatial distribution, region-specific proteins on a tumor lesion could be identified by protein extraction from tumor tissue and analysis using liquid chromatography with tandem mass spectrometry (LC-MS/MS). Using all the spectral data at each spot, various intensities of a specific peak were detected in the tumor and normal regions of the thyroid. Differences in the molecular weights of expressed proteins between tumor and normal regions were analyzed using unsupervised and supervised clustering. To verify the presence of discovered proteins through IMS, we identified ribosomal protein P2, which is specific for cancer. We have demonstrated the feasibility of IMS as a useful tool for the analysis of tissue sections, and identified the tumor-specific protein ribosomal protein P2.

A Case of Epstein-Barr Virus Associated Hemophagocytic Lymphohistiocytosis after Scrub Typhus Infection / 임상소아혈액종양

A 7-year-old boy with pancytopenia, cervical lymphadenopathy, interstitial pneumonia, and hepatosplenomegaly was diagnosed with Epstein-Barr Virus (EBV)-associated hemophagocytic lymphohistiocytosis. His clinical course was characterized by hepatorenal syndrome and myocarditis. Based on his serological markers for EBV and an immunochromatography test for scrub typhus, this case was inferred as an EBV infection that was reactivated during tsutsugamushi infection. We treated this patient with the HLH-2004 protocol and administered clarithromycin. Normal ferritin level was achieved within 8 weeks after starting chemotherapy and antibiotics.

Plasma Real Time-Quantitative Polymerase Chain Reaction of Epstein-Barr Virus in Immunocompetent Patients with Hepatitis / 대한소아소화기영양학회지

PURPOSE: Epstein-Barr virus (EBV) hepatitis is a usually asymptomatic and self-limiting disease in immunocompetent patients. However, the range of severity is wide, and the serological diagnosis is typically difficult until the convalescent phase. Thus, we examined the value of plasma EBV DNA real-time quantitative polymerase chain reaction (RT-qPCR) in EBV hepatitis for the timely diagnosis and the relationship between EBV viral load and clinical severity. METHODS: Sixty samples were confirmed as having EBV infection by RT-qPCR with the EBV BALF5 gene sequence. We examined the clinical characteristics of EBV hepatitis by reviewing medical records. RESULTS: The median total duration of fever was 8 days (range: 0-13 days). The mean peak value of aspartate aminotransferase (AST) was 241+/-214 U/L, and the mean peak value of alanine aminotransferase (ALT) was 298+/-312 U/L. There was no correlation between the serum levels of liver enzyme and plasma EBV DNA titer (p=0.1) or between median total duration of fever and EBV DNA titer (p=0.056). The median age of the EBV VCA IgM-negative group was lower compared with the EBV VCA IgM-positive group in EBV hepatitis (2 years vs. 6 years, p=0.0009). CONCLUSION: The severity of EBV hepatitis does not correlate with circulating EBV DNA load according to our data. Furthermore, we suggest that plasma EBV PCR may be valuable in young infants in whom the results of serology test for EBV infection commonly are negative.

A Case of Epstein-Barr Virus Associated Hemophagocytic Lymphohistiocytosis after Scrub Typhus Infection / 임상소아혈액종양

A 7-year-old boy with pancytopenia, cervical lymphadenopathy, interstitial pneumonia, and hepatosplenomegaly was diagnosed with Epstein-Barr Virus (EBV)-associated hemophagocytic lymphohistiocytosis. His clinical course was characterized by hepatorenal syndrome and myocarditis. Based on his serological markers for EBV and an immunochromatography test for scrub typhus, this case was inferred as an EBV infection that was reactivated during tsutsugamushi infection. We treated this patient with the HLH-2004 protocol and administered clarithromycin. Normal ferritin level was achieved within 8 weeks after starting chemotherapy and antibiotics.