Effects of Corticosteroids on Critically Ill Pulmonary Tuberculosis Patients With Acute Respiratory Failure: A Propensity Analysis of Mortality.

BACKGROUND: We investigated the effects of corticosteroids on the 90-day mortality outcomes in patients with pulmonary tuberculosis who were admitted to the intensive care unit (ICU) because of acute respiratory failure (ARF). METHODS: The medical records of 124 patients who had pulmonary tuberculosis with ARF and were admitted to the ICU at our tertiary referral center in South Korea between March 1989 and December 2014 were retrospectively analyzed. The 90-day mortality rate in this population was analyzed after adjustments with the inverse probability of treatment weighted (IPTW) method. RESULTS: The mean patient age was 62 years, and the 90-day mortality rate was 49.2% (61/124). Adjuvant steroids were used in 70 (56.5%) patients. The 90-day mortality rate was similar irrespective of corticosteroid use (48.6%, steroid group; 50.0%, nonsteroid group). The use of adjuvant steroids was not associated with the unadjusted 90-day mortality (odds ratio [OR], 0.94; 95% confidence interval [CI], .46-1.92; P = .875). In a comparison using an adjusted IPTW approach of the 90-day mortality between the 2 groups, we found that corticosteroid use was independently associated with reduced 90-day mortality (OR, 0.47; 95% CI, .22-.98; P = .049). CONCLUSIONS: The study results showed that corticosteroids could reduce the 90-day mortality rate in critically ill pulmonary tuberculosis patients with ARF.

Impact of Urea on Monoclonal Antibodies: Multiple Destabilization and Aggregation Effects for Therapeutic Immunoglobulin G Proteins.

We performed nano differential scanning fluorimetry (nanoDSF) measurements of immunoglobulin G (IgG) in urea gradient solutions under thermal unfolding. Our results show that the denaturing effect of urea on individual IgG domains can be monitored via a linear mapping of thermal shift curves to the corresponding urea concentrations. Assignment of IgG domains to each thermal shift curve allows for a reliable differentiation of the underlying mechanisms. Further results show a decisive influence of salt-induced electrostatic screening effects. We are able to explain all findings by preferential binding mechanisms in combination with electrostatic effects. The results of our study shed more light on the complex interaction mechanisms between buffer solutions and complex proteins, which are important for improving the shelf life of protein therapeutic formulation.

Development of biomarkers to distinguish different origins of red seabreams (Pagrus major) from Korea and Japan by fatty acid, amino acid, and mineral profiling.

Red seabream (Pagrus major) has been one of the most popular fish in East Asia since early times. However, the discharge of nuclear wastewater into the sea following the Fukushima nuclear disaster in Japan has led to violations of the country of origin labeling. Therefore, the aim of the present study was to determine the origin of fish based on fatty acid, amino acid, and mineral analyses, and to develop biomarkers that can discriminate between Japanese and Korean red seabream. To identify the differences between the two groups, 29 fatty acid families, 17 amino acids, and 4 minerals were analyzed in 60 fish samples (standard sample collected in autumn), and fatty acid profiles were analyzed using heatmap with hierarchical clustering analysis and orthogonal projections to latent structures discriminant analysis. The top 10 fatty acids that were different between the two groups were selected from all seasonal fish samples by combining variable importance in projection scores and p-values. According to the receiver operating characteristic curve analysis results, we proposed percentage linoleic acid (C18:2n-6, cis) as a candidate biomarker with excellent sensitivity and specificity. This study introduces a strategy to identify the origins of red seabream using linoleic acid obtained from fatty acid analysis.

Development of metabolomic biomarkers to discriminate the geographical origin of Korean and Russian snow crabs using CE-TOF/MS.

The quantity of snow crabs (Chionoecetes opilio) harvested in Korea is subject to seasonal restrictions; therefore, snow crabs are imported from Russia. Metabolites in snow crabs from two geographic origins were compared. The metabolites were subjected to metabolomic analysis to prevent fraudulent sales of marine products from a particular country. Capillary electrophoresis time-of-flight mass spectrometry was used. Seventy-seven target metabolites were identified using a mass spectral library. Through orthogonal partial least squares discriminant analysis, the top 25 biomarker candidates were evaluated based on p-values and fold changes. A total of 246 peaks (187 and 59 in the cation and anion modes, respectively) were identified. Among the biomarker candidates, 2-oxovaleric acid, asymmetric dimethylarginine, hypotaurine, and allo-threonine were selected as final biomarkers to unequivocally determine the geographic origin. Overall, metabolic analyses allowed us to differentiate snow crabs from different geographic origins. This method could also be extended of other marine products.

Perception of Long-Term Oxygen Therapy for Chronic Lung Diseases May Affect Poor Adherence in Korea.

BACKGROUND: Long-term oxygen therapy (LTOT) improves the survival of patients with hypoxemia due to chronic respiratory diseases. The clinical outcomes of LTOT are strongly associated with patient adherence. To improve the adherence of patients, physicians have focused on the efficacy of LTOT. However, poor adherence may stem from patients' perceptions of LTOT. Herein we evaluated patients' perceptions of LTOT affecting adherence. METHODS: We conducted a cross-sectional survey study using descriptive, open, and closed-ended questionnaire. Patients using oxygen therapy (OT) or requiring it but avoiding OT responded to the questionnaires at three university hospitals. RESULTS: Seventy-nine patients responded to the questionnaires. The number of patients using home and portable OT was 69 (93%) and 37 (46.3%), respectively. Patients with good adherence were 22 (30.1%). Among patients with good adherence, 90.9% used oxygen according to physicians' prescriptions whereas only 37.3% of those with poor adherence followed physicians' prescriptions (p<0.01). The reasons for avoiding using home OT were fear of permanent use (50%), unwanted attention (40%), and lack of symptoms (40%). They avoided portable OT because of unwanted attention (39%), heaviness (31.7%), and lack of symptoms (21.6%). CONCLUSION: Patients on LTOT had the perception of the misunderstanding the effects of OT and of psychosocial barriers to initiate or use LTOT. Considering these findings, health professionals need to provide effective education on the purpose of LTOT to improve patient adherence to OT and provide sufficient support for the management of psychosocial barriers in patients using LTOT.

Potential metabolomic biomarkers for evaluation of adriamycin efficacy using a urinary 1H-NMR spectroscopy.

A metabolomics approach using proton nuclear magnetic resonance (NMR) was applied to investigate metabolic alterations following adriamycin (ADR) treatment for gastric adenocarcinoma. After BALB/c-nu/nu mice were implanted with human gastric adenocarcinoma, ADR (1 or 3 mg kg(-1) per day) was intraperitoneally administered for 5 days. Urine was collected on days 2 and 5 and analyzed by NMR. The levels of trimethylamine oxide (TMAO, ×0.3), hippurate (×0.3) and taurine (×0.6) decreased significantly (P < 0.05), whereas the levels of 3-indoxylsulfate (×12.6), trigonelline (×1.5), citrate (×2.5), trimethylamine (TMA, ×2.0) and 2-oxoglutarate (×2.3) increased significantly (P < 0.05) in the tumor model. After ADR treatment, TMAO, hippuarte and taurine were increased significantly on day 5 compared with those of the tumor model. The levels of 2-oxoglutarate, 3-indoxylsulfate, trigonelline, TMA and citrate, which increased in the tumor model, significantly decreased to those of normal control by ADR treatment. Furthermore, the ratio between TMA and TMAO was dramatically altered in both tumor and ADR-treated groups. Overall, metabolites such as TMAO, TMA, 3-indoxylsulfate, hippurate, trigonelline, citrate and 2-oxoglutarate related to the tricarboxylic acid (TCA) cycle might be considered as therapeutic targets to potentiate the efficacy of ADR. Thus, these results suggest that the metabolomics analysis of tumor response to ADR treatment may be applicable for demonstrating the efficacy of anticancer agent, ADR and treatment adaptation.

Biomarker Development for Identifying Mud Loach (Misgurnus mizolepis) Origin Country Using Untargeted Metabolite Profiling.

Mud loach (Misgurnus mizolepis) has long been consumed in Korea. Recently, Chinese mud loaches were replaced with expensive Korean mud loaches, owing to taste and preference. Such issues occur in aquatic food distribution processes, leading to inferior food delivery. Previously, a study was conducted to confirm the origin of mud loaches using genetic analysis. However, untargeted metabolites profiling of mud loaches has not been reported. Untargeted metabolomics provides information on the overall metabolic profiling of a sample, allowing the identification of new metabolites. Here, we analyzed the metabolites of mud loaches of different geographical origins using liquid chromatography (LC)-quadrupole-time-of-flight mass spectrometry (MS). Orthogonal partial least squares discriminant analysis from LC/MS datasets showed a clear distinction between Korean and Chinese mud loaches, and univariate statistical analysis showed significantly different metabolites between them. N-acetylhistidine and anserine were selected as biomarkers for geographical origin discrimination using the receiver operating characteristic curve. N-acetylhistidine and anserine levels were significantly higher in Chinese than in Korean mud loaches. These results indicate that metabolic analysis can be used to discriminate between the geographical origins of mud loaches, curtailing the inadvertent substitution of mud loaches from different regions.

Characterization of the complete mitochondrial genome of demersal flatfish Eopsetta grigorjewi (Pleuronectiformes: Pleuronectidae) from South Korea.

Eopsetta grigorjewi (Pleuronectiformes: Pleuronectidae) is a demersal flatfish found in South Korea, Japan, Taiwan, China, and the Yellow Sea. E. grigorjewi complete mitochondrion DNA (mtDNA) consists of 16,921 bp and a 54% A + T content. It includes 2 ribosomal RNA (rRNA), 22 transfer RNA (tRNA), 13 protein-coding genes, and 1 non-coding regulatory area. ND2, ND3, ND4, COII, COIII, ATPase6, and CytB all have incomplete stop codon genes. The evolutionary analysis of 13 species from the same family indicated a close relationship. This work will be valuable for future research on molecular evolution and the creation of biomarker databases for determining the originality of E. grigorjewi.

Complete mitochondrial genome of marine Petrale sole Eopsetta jordani (Pleuronectiformes: Pleuronectidae) flatfish.

Petrale sole Eopsetta jordani (Pleuronectiformes: Pleuronectidae) is a species of flounder, found in the northeastern Pacific Ocean and the Bering Sea of the United States and Canada. The complete mitochondrial DNA (mtDNA) of E. jordani has 16,483 bp with an overall A + T content of 61% and consists of 2 ribosomal RNA (rRNA) genes, 13 protein-coding genes (PCGs), 22 transfer RNA (tRNA) genes, and a non-coding control region. It has incomplete stop codon genes in ND2, COII, ATPase6, COIII, ND3, and ND4. Phylogenetic analysis indicated that E. jordani is not monophyletic with cogeneric Eopsetta grigorjewi and is separated from other species in the same family by a large distance. Present study results provide useful data for further research on genetic diversity and evolution of the Eopsetta and the Pleuronectidae.

Complete mitochondrial genome of scorpionfish Scorpaena neglecta (Actinopterygii).

Scorpaena neglecta (Temminck and Schlegel, 1843) is a marine fish, in the family Scorpaenidae, order Scorpaeniformes, class Actinopterygii of the phylum Chordata. The first species of Scorpaena with a complete mitochondrial genome is described in the present study. The circular mitochondrial genome of S. neglecta has 17,202 bp with 54.75% A + T content and encodes 37 genes, including 13 protein-coding genes (PCGs), 22 transfer RNA (tRNA), and two ribosomal RNA (rRNA). The phylogenetic tree indicates S. neglecta clustered into one branch and is closely related to other Scorpaenidae species. The mitochondrial genome structure and gene content of S. neglecta will support the study of evolution and phylogenetic relationships among Scorpaenidae species.

Ethyl Carbamate Formation from Cyanate in Model System of Ethanol-Water Media Using Response Surface Methodology.

Ethyl carbamate (EC) has been identified as a possible human carcinogen belonging to Group 2A. EC is naturally formed during the fermentation and storage of alcoholic drinks and fermented foods. When ingested in large amounts, EC can cause various health problems, such as gastroenteric hemorrhage, vomiting, and cancer. In this study, optimization of EC formation from cyanate was examined using response surface methodology (RSM), a central composite design that includes variables such as alcohol concentration (10, 15, 20, 25, and 30%), pH (2.5, 3.0, 3.5, 4.0, and 4.5), storage temperature (5, 10, 15, 20, and 25°C), and storage duration (2, 4, 6, 8, and 10 days). EC content was determined using gas chromatography with flame ionization detection and the results were optimized using RSM. EC formation from cyanate degradation was found to increase with storage duration and temperature, acidity, and alcohol concentration. Cy-anate degradation was associated with the formation of EC. Approximately 83.1±0.1% of cyanate was degraded to 538±9 µM of EC. However, not all of the cyanate reacted with ethanol during fermentation to form EC. This study aimed to develop the ideal conditions for EC analysis to reduce EC production in alcoholic drinks and fermented foods.

Comparison of Extraction Methods and Extraction Optimization in Aqueous Two-Phase System for Ethyl Carbamate Analysis in Model Systems for Maesil Wine.

Ethyl carbamate (EC) is a known carcinogen, and therefore its intake is regulated internationally. The objectives of this study were to compare the EC recovery yields under different liquid-liquid extraction (LLE) conditions and to investigate the optimum conditions of the aqueous two-phase system (ATPS) for EC extraction. Our results showed that for the LLE method, addition of 15% NaCl improved the EC yield by 15%, and dichloromethane as the extraction solvent showed a slightly higher yield (about 5%) than chloroform. However, there was little difference in the yield when mixing was performed using an ultrasonic bath compared to a vortex mixer. Using response surface methodology with central composite design to analyze the ATPS results, optimal extraction was found to occur at 21.5°C for 2.8 h in the sample containing 70% alcohol and 15% phosphate, showing a recovery yield of 75.64%. This information can be applied to alcoholic beverages and other fermented food products to analyze EC with better extraction methods, depending on the types of food.

(1)H NMR-based metabolomic profiling in mice infected with Mycobacterium tuberculosis.

Tuberculosis (TB) is one of three major infectious diseases, and the control of TB is becoming more difficult because of the emergence of multidrug-resistant and extensively drug-resistant strains. In this study, we explored the (1)H NMR-based metabolomics of TB using an aerobic TB infection model. Global profiling was applied to characterize the responses of C57Bl/6 mice to an aerobic infection with virulent Mycobacterium tuberculosis (MTB). The metabolic changes in organs (i.e., the lung, the target organ of TB, and the spleen and liver, remote systemic organs) and in serum from control and MTB-infected rats were investigated to clarify the host-pathogen interactions in MTB-infected host systems. Principal components analysis (PCA) and orthogonal partial least-squares discriminant analysis (OPLS-DA) score plots showed distinct separation between control and MTB-infected rats for all tissue and serum samples. Several tissue and serum metabolites were changed in MTB-infected rats, as compared to control rats. The precursors of membrane phospholipids, phosphocholine, and phosphoethanolamine, as well as glycolysis, amino acid metabolism, nucleotide metabolism, and the antioxidative stress response were altered based on the presence of MTB infection. This study suggests that NMR-based global metabolite profiling of organ tissues and serum could provide insight into the metabolic changes in host infected aerobically with virulent Mycobacterium tuberculosis.

Complete mitochondrial genome of sea peach Halocynthia aurantium (stolidobranchia: Pyuridae) from Korea.

Halocynthia aurantium (Stolidobranchia: Pyuridae) is a species of tunicate of commercial value that is commonly found in the northern Pacific Ocean and in the Bering Sea. Here, we determined the complete mitogenome of sea peach H. aurantium using 150 PE high-throughput sequencing. The assembled mitogenome is 14,979 bp in length (overall A + T contents 56.2%), and contains 13 protein-coding genes, 21 transfer RNAs, two ribosomal RNAs. Phylogenetic analysis of the mitogenome sequence of H. aurantium fully resolved it in a clade with H. roretzi. These data and results will be useful for future studies on the evolution of the Halocynthia and the Pyuridae.

Development and validation of multiplex PCR assay for differentiating tunas and billfishes.

Commercially available tunas and billfishes are generally processed as steaks, making it difficult to visually distinguish between the two. We developed and validated species-specific primers to prevent the adulteration of tunas by billfishes. Tunas and billfishes primers were designed on the cytochrome oxidase subunit I. Multiplex PCR bands obtained were 579 bp, 291 bp and 114 bp for tunas, billfishes and internal control. Sensitivity was determined to be 5 ng for tunas and billfishes. A total of 50 samples were monitored: 49 for tunas and 1 for billfish. As a result of the monitoring, the fake tunas did not show due to the agreement between product name and the raw material of the wrapping paper. Our results indicate that the species-specific primers developed in this study are suitable for differentiating tunas and billfishes. The newly developed multiplex PCR assay is a time and cost effective technique for determining the authenticity of tunas and billfishes.

Metabolic profiling of kidney and urine in rats with lithium-induced nephrogenic diabetes insipidus by (1)H-NMR-based metabonomics.

Lithium (Li) treatment for bipolar affective disorders is associated with a variety of renal side effects. The metabolic response of the kidney to chronic Li treatment has rarely been studied. We applied a novel method of (1)H-nuclear magnetic resonance (NMR)-based metabonomics to integrate metabolic profiling and to identify the changes in the levels of metabolites in the kidney and urine from rats with Li-induced NDI. Metabolic profiles of urine and kidney homogenate [3 different zones (cortex, outer medulla, and inner medulla) or whole kidney] were investigated using high-resolution NMR spectroscopy coupled with pattern recognition methods. The accurate concentrations of metabolites in kidney homogenates and urine were rapidly measured using the target-profiling procedure, and the difference in the levels of metabolites was compared using multivariate analysis, such as principal component analysis and orthogonal partial least squares-discriminant analysis. Major endogenous metabolites for kidney homogenates contained products of glycolysis (glucose, lactate) and amino acids, as well as organic osmolytes (e.g., betaine, myo-inositol, taurine, and glycerophosphocholine). Many metabolites revealed changes in their levels, including decreased levels of organic osmolytes and amino acids in the inner medulla. A number of urinary metabolites were changed in Li-induced NDI, and in particular, elevated urinary levels of acetate, lactate, allantoin, trimethylamine, and creatine could suggest Li-induced renal cell stress or injury. Taken together, metabonomics of kidney tissue and urine based on (1)H-NMR spectroscopy could provide insight into the effects of Li-induced renal effects and cell injury.

Toxicometabolomics of urinary biomarkers for human gastric cancer in a mouse model.

Toxicometabolomics of urinary biomarkers for human gastric cancer in a mouse model was investigated using (1)H-nuclear magnetic resonance (NMR) spectroscopy. A human gastric adenocarcinoma cell line (1 × 10(7) cells/ml) was grafted onto the skin of the back of intact male BALB/c-nu/nu mice. After the xenografted tumors developed, urine was collected and analyzed for endogenous metabolites. Global profiling combined with principal components analysis (PCA), partial least squares-discriminant analysis (PLS-DA), and orthogonal projections to latent squares-discriminant analysis (OPLS-DA) showed distinct separation of clusters between control and tumor-bearing mice. Targeted profiling revealed significant changes in trimethylamine oxide (TMAO), 3-indoxylsulfate, hippurate, and citrate levels in mice carrying human gastric cancer cells compared to normal mice. The levels of TMAO (0.41-fold) and hippurate (0.26-fold) in tumor-bearing mice were significantly decreased, whereas the levels of 3-indoxylsulfate (3.39-fold), 2-oxoglutarate (2.32-fold), and citrate (1.9-fold) were significantly increased in urine samples of tumor-bearing mice. Data suggest that TMAO, hippurate, 3-indoxylsulfate, 2-oxoglutarate, and citrate may serve as useful urinary biomarkers for gastric tumorigenesis in a mouse model.

The complete mitochondrial genome of pitted stingray Bathytoshia brevicaudata (Myliobatiformes: Dasyatoidea).

The complete mitochondrial genome of pitted stingray, Bathytoshia brevicaudata (Myliobatiformes: Dasyatoidea) was investigated by next-generation sequencing. The analyzed mitochondrial genome was 17,640 nucleotides in length and had 59.2% for AT contents. This genome contains 2 ribosomal RNA genes, 13 protein-coding genes, 22 transfer RNA genes. and 1 putative control region. Five protein-coding genes (ATPase6, COII, ND2, ND3, ND4) including incomplete stop codons and four tRNAs have atypical codons. The phylogenetic inference including 13 species of the same family revealed a close relationship with Pteroplatytrygon violacea. This is the first mitochondrial genome report from genus Bathytoshia.

A metabonomic study on the biochemical effects of doxorubicin in rats using (1)H-NMR spectroscopy.

Metabonomic investigation of doxorubicin (adriamycin) was carried out in male Sprague-Dawley rats using high-resolution (1)H nuclear magnetic resonance (NMR) spectroscopy coupled with multivariate statistics. Urine samples (d -1 to 7) from rats treated with doxorubicin at two dose levels (5 or 15 mg/kg body weight) were collected at each time point and doxorubicin-induced biomarkers were examined. Of metabolites, early elevated biochemical changes were observed in trimethylamine N-oxide (TMAO) levels suggesting renal dysfunction. Perturbation in TMAO was maximal in the low-dose group at 48 h post dose (p.d.) and returned to control at 168 h p.d., indicating recovery from renal toxicity induced by doxorubicin. After doxorubicin administration, the high-dose group was divided into low and high responders at 48 h and further divided into high, moderate, and no recovery animals at 96 h, indicating individual susceptible response to drug-induced toxicity. Urinary increases in glucose, lactate, alanine, and valine suggested progression of renal damage resulting in glycosuria, lactic aciduria, and aminoaciduria up to 168 h in the high-dose group. Urinary elevation of creatine and phenylacetylglycine (PAG) together with reduction of N-methylnicotinic acid (NMNA) and hippurate levels was suggestive of liver injury in the high-dose group. Impairment of energy metabolism was also indicated by decreased levels of tricarboxylic acid cycle intermediates in urine of rats treated with high-dose doxorubicin. This study highlights the applicability of NMR-based metabonomics with multivariate statistics for monitoring biomarkers produced by doxorubicin treatments.