Aims: This study was aimed to express Meyerozyma guilliermondii strain RT lipase using Komagataella phaffii X-33 expression system and its biochemical characterization and analyse the predicted structure of the product. Methodology and results: Meyerozyma guilliermondii strain RT obtained from the previous study was used as the source of RT lipase gene. Extracellular M. guilliermondii strain RT lipase expression has significantly been improved up to 56 U/mg at 24 h cultivation in Yeast extract-Peptone-Dextrose (YPD) medium containing (in w/v): 1% yeast extract, 2% peptone, 2% dextrose with 0.5% v/v methanol induction. Characterization of RT lipase showed optimum activity at 45 °C and pH 9. It exhibited stability in the alkaline pH range (8 to 10) and retained 50% of its residual activity at 30 °C for 30 min. Substrate specificity analysis revealed that it preferred short to medium-chain triacylglycerols (C2-C12) with the highest activity towards caprylic acid (C8). Pairwise alignment revealed three substitutions (S2L, S92L and S193L) present in non-CTG-clade hosts (K. phaffii). Homology modelling (YASARA) was used to predict the structures of RT lipase [wild type (wt) and recombinant (rc)]. Mutational analysis of the structures showed the differences in loops that might attribute to the reduction of the optimum temperature from 75 °C (wt) to 45 °C (rc). Conclusion, significance and impact of study RT lipase was successfully overexpressed extracellularly using K. phaffii expression system with 91.8-fold higher specific activity than the native host. The conceptual advances on the importance of codon optimization before expressing a protein from a CTG-clade species in a non-CTG-clade yeast have been highlighted and the effect of the rare codon usage in recombinant protein characteristics has been evident.
Candida ; Lipase--analysis

Human breast cancer cell line, MDA-MB-231, is highly invasive and aggressive, compared to less invasive cell line, MCF-7. To explore the genes that might influence the malignancy of MDA-MB-231, DNA microarray analysis was performed. The results showed that G0/G1 switch 2 (G0S2) was one of the most highly expressed genes among the genes upregulated in MDA-MB-231. Although G0S2 acts as a direct inhibitor of adipose triglyceride lipase, action of G0S2 in cancer progression is not yet understood. To investigate whether G0S2 affects invasiveness of MDA-MB-231 cells, G0S2 expression was inhibited using siRNA, which led to decreased cell proliferation, migration, and invasion of MDA-MB-231 cells. Consequently, G0S2 inhibition inactivated integrin-regulated FAK-Src signaling, which promoted Hippo signaling and inactivated ERK1/2 signaling. In addition, G0S2 downregulation decreased β-catenin expression, while E-cadherin expression was increased. It was demonstrated for the first time that G0S2 mediates the Hippo pathway and induces epithelial to mesenchymal transition (EMT). Taken together, our results suggest that G0S2 is a major factor contributing to cell survival and metastasis of MDA-MB-231 cells.
Breast Neoplasms ; Breast ; Cadherins ; Cell Line ; Cell Proliferation ; Cell Survival ; Down-Regulation ; Humans ; Lipase ; Neoplasm Metastasis ; Oligonucleotide Array Sequence Analysis ; RNA, Small Interfering ; Signal Transduction

PURPOSE: Glyphosate herbicides (GHs) are widely used and increasingly associated with poisoning cases. Acute pancreatitis (AP) is among the many complications associated with the toxicity of GHs. We investigated the relationship between incidence of AP and its prognosis in patients with GH poisoning. METHODS: This was a retrospective cohort study conducted at a single tertiary hospital between January 2004 and December 2014. We enrolled all patients presented to the emergency department with GH poisoning. The Clinical and laboratory variables were analyzed to investigate the relationship between GH intoxication and AP. RESULTS: We studied 245 patients. Incidence of AP after GH poisoning was 6.5%. Patients with AP (mean 66 years) were older than the non-AP group (56 years). Systolic blood pressure, Glasgow Coma Scale, and amount of ingested poison differed significantly between the two groups. In the blood tests, white blood cell count, alanine aminotransferase, glucose, potassium, amylase, and lipase showed significant differences. The pH, bicarbonate, and lactate levels also differed significantly. Patients with AP demonstrated higher incidence of respiratory failure, pneumonia, acute kidney injury, rhabdomyolysis, and intensive care unit stay time. Additionally, 30-day mortality (n=11, 68.8%) was significantly higher in the AP group. On multivariate analysis, adjusted age, amount of ingestion, and lactate correlated with occurrence of AP. CONCLUSION: The incidence of GH-induced AP was 6.5% with a 30-day mortality of 68.8%. The patient's age, ingested dosage, and lactate levels were associated with GH-induced AP.
Acute Kidney Injury ; Alanine Transaminase ; Amylases ; Blood Pressure ; Cohort Studies ; Eating ; Emergency Service, Hospital ; Glasgow Coma Scale ; Glucose ; Hematologic Tests ; Herbicides ; Humans ; Hydrogen-Ion Concentration ; Incidence ; Intensive Care Units ; Lactic Acid ; Leukocyte Count ; Lipase ; Mortality ; Multivariate Analysis ; Pancreatitis* ; Pneumonia ; Poisoning ; Potassium ; Prognosis* ; Respiratory Insufficiency ; Retrospective Studies ; Rhabdomyolysis ; Tertiary Care Centers

Chylomicronemia is a severe type of hypertriglyceridemia characterized by chylomicron accumulation that arises from a genetic defect in intravascular lipolysis. It requires urgent and proper management, because serious cases can be accompanied by pancreatic necrosis or persistent multiple organ failure. We present the case of a 1-month-old infant with chylomicronemia treated by plasmapheresis. His chylomicronemia was discovered incidentally when lactescent plasma was noticed during routine blood sampling during a hospital admission for fever and irritability. Laboratory investigation revealed marked triglyceridemia (>5,000 mg/dL) with high chylomicron levels. We therefore decided to perform a therapeutic plasmapheresis to prevent acute pancreatitis. Sequence analysis revealed a homozygous novel mutation in exon 4 of GPIHBP1: c.476delG (p.Gly159Alafs). Glycosylphosphatidylinositol-anchored high density lipoprotein-binding protein 1 (GPIHBP1) stabilizes the binding of chylomicrons near lipoprotein lipase and supports lipolysis. Mutations of GPIHBP1, the most recently discovered gene, can lead to severe hyperlipidemia and are known to make up only 2% of the monogenic mutations associated with chylomicronemia. The patient maintains mild hypertriglyceridemia without rebound after single plasmapheresis and maintenance fibrate medication so far. Here, we report an infant with chylomicronemia due to GPIHBP1 mutation, successfully treated by plasmapheresis.
Chylomicrons ; Exons ; Fever ; Humans ; Hyperlipidemias ; Hypertriglyceridemia ; Infant* ; Infant, Newborn* ; Lipolysis ; Lipoprotein Lipase ; Multiple Organ Failure ; Necrosis ; Pancreatitis ; Plasma ; Plasmapheresis* ; Sequence Analysis

Drug-induced liver injury (DILI) is the serious and fatal drug-associated adverse effect, but its incidence is very low and individual variation in severity is substantial. Acetaminophen (APAP)-induced liver injury accounts for >50% of reported DILI cases but little is known for the cause of individual variations in the severity. Intrinsic genetic variation is considered a key element but the identity of the genes was not well-established. Here, pre-biopsy method and microarray technique was applied to uncover the key genes for APAP-induced liver injury in mice, and a cause and effect experiment employing quantitative real-time PCR was conducted to confirm the correlation between the uncovered genes and APAP-induced hepatotoxicity. We identified the innately and differentially expressed genes of mice susceptible to APAP-induced hepatotoxicity in the pre-biopsied liver tissue before APAP treatment through microarray analysis of the global gene expression profiles (Affymetrix GeneChip® Mouse Gene 1.0 ST for 28,853 genes). Expression of 16 genes including Gdap10, Lpl, Gabra3 and Ccrn4l were significantly different (t-test: FDR <10%) more than 1.5 fold in the susceptible animals than resistant. To confirm the association with the susceptibility to APAP-induced hepatotoxicity, another set of animals were measured for the expression level of selected 4 genes (higher two and lower two genes) in the liver pre-biopsy and their sensitivity to APAP-induced hepatotoxicity was evaluated by post hoc. Notably, the expressions of Gabra3 and Lpl were significantly correlated with the severity of liver injury (p<0.05) demonstrating that these genes may be linked to the susceptibility to APAP-induced hepatotoxicity.
Acetaminophen ; Animals ; Drug-Induced Liver Injury ; Genetic Variation ; Incidence ; Lipoprotein Lipase* ; Lipoproteins* ; Liver ; Methods ; Mice ; Microarray Analysis ; Real-Time Polymerase Chain Reaction ; Receptors, GABA-A* ; Toxicogenetics ; Transcriptome

BACKGROUND/AIMS: Non-pancreatic elevations of serum lipase have been reported, and differential diagnosis is necessary for clinical practice. This study aimed to evaluate the clinical efficacy of serum lipase subtype analysis for the differential diagnosis of pancreatic and non-pancreatic lipase elevation. METHODS: Patients who were referred for the serum lipase elevation were prospectively enrolled. Clinical findings and serum lipase subtypes were analyzed and compared by dividing the patients into pancreatitis and non-pancreatitis groups. RESULTS: A total of 34 patients (12 pancreatitis vs. 22 non-pancreatitis cases) were enrolled. In univariate analysis, the fraction of pancreatic lipase (FPL) in the total amount of serum lipase subtypes was statistically higher in patients with pancreatitis ([median, 0.004; interquartile range [IQR], 0.003 to 0.011] vs. [median, 0.002; IQR, 0.001 to 0.004], p = 0.04). Based on receiver operating characteristic curve analysis for the prediction of acute pancreatitis, FPL was the most valuable predictor (area under the receiver-operating characteristic curve [AUROC], 0.72; 95% confidence interval [CI], 0.54 to 0.86; sensitivity, 83.3%; specificity, 63.6%; positive predictive value, 55.6%; negative predictive value, 97.5%). In multivariate analysis, a cut-off value higher than 0.0027 for the FPL was associated with acute pancreatitis (odds ratio, 8.3; 95% CI, 1.3 to 51.7; p = 0.02). CONCLUSIONS: The results did not support that serum lipase subtype analysis could replace standard lipase measurement for the diagnosis of acute pancreatitis. However, the test demonstrated adequate sensitivity for use in triage or as an add-on test for serum lipase elevation.
Diagnosis ; Diagnosis, Differential* ; Humans ; Lipase* ; Multivariate Analysis ; Pancreatitis ; Prospective Studies ; ROC Curve ; Sensitivity and Specificity ; Treatment Outcome* ; Triage

Chronic hepatitis C virus (HCV) infection is a major cause of liver cirrhosis and hepatocellular carcinoma. Combination therapy of pegylated interferon-alpha (PEG-IFN-α) and ribavirin (RBV) is a current standard treatment for chronic HCV infection in Korea, which has considerable adverse effects. Acute pancreatitis is a rare complication of PEG-IFN-α administration. We report a case of a 62-year-old female who experienced acute pancreatitis after 4 weeks of PEG-IFN-α-2a and RBV combination therapy for chronic HCV infection. The main cause of the acute pancreatitis in this case was probably PEG-IFN-α rather than RBV for several reasons. A few cases have been reported in which acute pancreatitis occurred during treatment with PEG-IFN-α-2b. This is the first report of acute pancreatitis associated with PEG-IFN-α-2a in Korea.
Amylases/analysis ; Antiviral Agents/adverse effects/*therapeutic use ; Drug Therapy, Combination ; Female ; Hepatitis C, Chronic/diagnostic imaging/*drug therapy ; Humans ; Interferon-alpha/adverse effects/*therapeutic use ; Lipase/analysis ; Middle Aged ; Pancreatitis/*etiology ; Polyethylene Glycols/adverse effects/*therapeutic use ; Recombinant Proteins/adverse effects/therapeutic use ; Republic of Korea ; Ribavirin/therapeutic use ; Tomography, X-Ray Computed

PURPOSE: The aim of the study was to determine the factors associated with complicated acute cholecystitis of initial clinical findings during an emergency department (ED) visit, and to use them as a guideline for consideration of early and active surgical intervention, to improve the prognosis of acute cholecystitis. METHODS: Medical records of adult patients diagnosed and treated in the ED as acute cholecystitis were reviewed retrospectively. Clinical findings including demographic data, past medical history, symptoms, physical exam, and laboratory test results were included in the analysis. A case associated with gall bladder empyema, gangrene, perforation, hydrops, or failure of initial laparoscopic approach was defined as complicated acute cholecystitis. Factors showing significance in univariate analyses were included in binary logistic regression analysis for prediction of complicated acute cholecystitis. RESULTS: Age, sex, hypertension history, anorexia, body temperature, white blood cell count (WBC), aspartate aminotransferase, creatinine, total bilirubin, amylase, and lipase were significant in univariate analyses, and included in multivariate analysis. Age (p=0.039), male sex (p=0.004), and WBC (p=0.019) were significant in multivariate analysis. CONCLUSION: Age, sex, and initial WBC of patients diagnosed and treated in the ED as acute cholecystitis were independently associated with complicated acute cholecystitis.
Adult ; Amylases ; Anorexia ; Aspartate Aminotransferases ; Bilirubin ; Body Temperature ; Cholecystitis ; Cholecystitis, Acute* ; Creatinine ; Edema ; Emergencies* ; Emergency Service, Hospital* ; Gangrene ; Humans ; Hypertension ; Leukocyte Count ; Lipase ; Logistic Models ; Male ; Medical Records ; Multivariate Analysis ; Prognosis ; Retrospective Studies

Fenofibrate is one of PPAR-alpha (peroxisome proliferator activated receptor alpha) agonists. Fenofibrate decreases effectively triglyceride and increases high density lipoprotein cholesterol level through the effect on lipoprotein lipase, hepatic production and degradation of lipoproteins. Fenofibrate was recommended as the drug for hypertriglyceridemia treatment in European guideline released in 2011. But American heart association guideline in 2013 did not recommend non-statin therapy including fibrate for the prevention of atherosclerotic cardiovascular disease. But fenofibrate is still considered as the important drug for the management of atherogenic dyslipidemia especially in patients with metabolic syndrome and diabetes to reduce the residual risk after statin therapy from the evidence of many studies. Fibrates including bezafibrate, gemfibrozil, and fenofibrate increased serum creatinine level in several studies. But the mechanism of change in renal function is not clear till now. And the reversibility of renal function with drug discontinuation is dependent on the kinds of fibrate. Fenofibrate increased serum creatinine level, decreased albuminuria and renal function was reversible with the drug discontinuation in large clinical trials. In these days renal function change with fenofibrate therapy in Korean patients with hypertriglyceridemia was investigated. Fenofibrate treatment for 2 months increased serum creatinine level significantly and old age was associated with the change of renal function in multivariate analysis. Short-term therapy significantly increased serum creatinine level even within normal range, and this change may be important in some groups especially old age.
Albuminuria ; American Heart Association ; Bezafibrate ; Cardiovascular Diseases ; Cholesterol, HDL ; Creatinine ; Dyslipidemias ; Fenofibrate* ; Fibric Acids ; Gemfibrozil ; Glomerular Filtration Rate ; Humans ; Hydroxymethylglutaryl-CoA Reductase Inhibitors ; Hypertriglyceridemia ; Lipoprotein Lipase ; Lipoproteins ; Multivariate Analysis ; Reference Values ; Triglycerides

PURPOSE: Carbamate insecticides are potent cholinesterase inhibitors capable of causing severe cholinergic toxicity. Use of carbamate rather than organophosphate insecticides has been increasing. Compared with organophosphate poisoning, relatively few studies have investigated carbamate-associated acute pancreatitis. We investigated general characteristics and pancreatitis of carbamate poisoning and the predictors, among those readily assessed in the emergency department. METHODS: We performed a retrospective review of consecutive patients, aged over 18 years, who were admitted between January 2008 and April 2012 to an emergency department (ED) of an academic tertiary care center for treatment of carbamate poisoning. Patients who exhibited poisoning by any other material, except alcohol, were excluded. After application of exclusion criteria, patients were divided according to carbamate-induced pancreatitis and non-pancreatitis groups. RESULTS: A total of 41 patients were included in this study. Among these 41 patients, the prevalence of acute pancreatitis was 36.6% (15 patients). Initial blood chemistry tests showed a statistically higher glucose level in the pancreatitis group, compared with the non-pancreatitis group (222, IQR 189-284 vs. 137, IQR 122-175 mg/dL, P<0.05). Regarding clinical courses and outcomes, a significantly higher proportion of patients developed pneumonia [10 (66.7%) vs. 6 (23.1%), P<0.05] and had a longer hospital stay (7 days, IQR 6-12 vs. 5 days, IQR 2-11, P<0.05), but no difference in mortality, in the pancreatitis group vs. the non-pancreatitis group. In multivariate analysis, the initial glucose was showing significant association with the presentation of carbamate-induced acute pancreatitis (odds ratio 1.018, 95% confidence interval 1.001-1.035, P<0.05). CONCLUSION: Carbamate-induced acute pancreatitis is common, but not fatal. Initial serum glucose level is associated with acute pancreatitis.
Blood Glucose ; Carbamates ; Chemistry ; Cholinesterase Inhibitors ; Emergency Service, Hospital ; Glucose ; Humans ; Insecticides ; Length of Stay ; Lipase ; Mortality ; Multivariate Analysis ; Organophosphate Poisoning ; Pancreatitis* ; Pneumonia ; Poisoning* ; Prevalence ; Retrospective Studies ; Tertiary Care Centers