BACKGROUND: Kidney ischemia-reperfusion (IR) via laparotomy is a conventional method for kidney surgery in a mouse model. However, IR, an invasive procedure, can cause serious acute and chronic complications through apoptotic and inflammatory pathways. To avoid these adverse responses, a Non-IR and dorsal slit approach was designed for kidney surgery. METHODS: Animals were divided into three groups, 1) sham-operated control; 2) IR, Kidney IR via laparotomy; and 3) Non-IR, Non-IR and dorsal slit. The effects of Non-IR method on renal surgery outcomes were verified with respect to animal viability, renal function, apoptosis, inflammation, fibrosis, renal regeneration, and systemic response using histology, immunohistochemistry, real-time polymerase chain reaction, serum chemistry, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining, and Masson's trichrome staining. RESULTS: The Non-IR group showed 100% viability with mild elevation of serum blood urea nitrogen and creatinine values at day 1 after surgery, whereas the IR group showed 20% viability and lethal functional abnormality. Histologically, renal tubule epithelial cell injury was evident on day 1 in the IR group, and cellular apoptosis enhanced TUNEL-positive cell number and Fas/caspase-3 and KIM-1/NGAL expression. Inflammation and fibrosis were high in the IR group, with enhanced CD4/CD8-positive T cell infiltration, inflammatory cytokine secretion, and Masson's trichrome stain-positive cell numbers. The Non-IR group showed a suitable microenvironment for renal regeneration with enhanced host cell migration, reduced immune cell influx, and increased expression of renal differentiation-related genes and anti-inflammatory cytokines. The local renal IR influenced distal organ apoptosis and inflammation by releasing circulating pro-inflammatory cytokines. CONCLUSION: The Non-IR and dorsal slit method for kidney surgery in a mouse model can be an alternative surgical approach for researchers without adverse reactions such as apoptosis, inflammation, fibrosis, functional impairment, and systemic reactions.
Animals ; Apoptosis ; Blood Urea Nitrogen ; Cell Count ; Cell Movement ; Chemistry ; Creatinine ; Cytokines ; DNA Nucleotidylexotransferase ; Epithelial Cells ; Fibrosis ; Immunohistochemistry ; Inflammation ; Kidney ; Laparotomy ; Methods* ; Mice* ; Nephrectomy* ; Real-Time Polymerase Chain Reaction ; Regeneration*

Diagnosis of the urea cycle disorder (USD) carbamoyl-phosphate synthetase 1 (CPS1) deficiency (CPS1D) based on only the measurements of biochemical intermediary metabolites is not sufficient to properly exclude other UCDs with similar symptoms. We report the first Korean CPS1D patient using whole exome sequencing (WES). A four-day-old female neonate presented with respiratory failure due to severe metabolic encephalopathy with hyperammonemia (1,690 µmol/L; reference range, 11.2-48.2 µmol/L). Plasma amino acid analysis revealed markedly elevated levels of alanine (2,923 µmol/L; reference range, 131-710 µmol/L) and glutamine (5,777 µmol/L; reference range, 376-709 µmol/L), whereas that of citrulline was decreased (2 µmol/L; reference range, 10-45 µmol/L). WES revealed compound heterozygous pathogenic variants in the CPS1 gene: one novel nonsense pathogenic variant of c.580C>T (p.Gln194*) and one known pathogenic frameshift pathogenic variant of c.1547delG (p.Gly516Alafs*5), which was previously reported in Japanese patients with CPS1D. We successfully applied WES to molecularly diagnose the first Korean patient with CPS1D in a clinical setting. This result supports the clinical applicability of WES for cost-effective molecular diagnosis of UCDs.
Base Sequence ; Carbamoyl-Phosphate Synthase (Ammonia)/chemistry/*genetics ; Carbamoyl-Phosphate Synthase I Deficiency Disease/diagnosis/*genetics ; Codon, Nonsense ; Exons ; Female ; Frameshift Mutation ; High-Throughput Nucleotide Sequencing ; Humans ; Infant, Newborn ; Republic of Korea ; Sequence Analysis, DNA ; Urea Cycle Disorders, Inborn/diagnosis

BACKGROUND: Prompt and accurate urine chemistry analysis is important to provide information for diagnosis and therapy. In this study, we evaluated the overall performance and utility of an automated chemistry analyser for urine chemistry testing in accordance with Clinical and Laboratory Standards Institute guidelines. METHODS: From January 2015 to March 2015, we evaluated the precision, linearity, limits of detection, carryover, and turnaround times after automation of nine items: total protein, albumin, glucose, blood urea nitrogen, total calcium, magnesium, inorganic phosphate, creatinine, and uric acid. A Hitachi 7600-110 instrument (Hitachi Ltd., Japan) and Hitachi ID Privileged Access Manager (Hitachi Ltd.) were used for automated chemistry analysis and sample preparation, respectively. RESULTS: Regarding precision, the coefficient of variation was 3.9% to 1.6% for high levels and 3.3% to 24.1% for low levels. The linearity and coefficients of determination of all the test items were acceptable. Performance comparison revealed that the two systems were comparable, as evidenced by correlation coefficients >0.975 for most items; moreover, carryover of all items was <1%. The mean turnaround time was 59 minutes. CONCLUSIONS: Urine chemistry testing can be performed with acceptable precision, linearity, and performance by using the Hitachi 7600-110 automated chemistry analyser. The sample preparation system reduces turnaround time, which enhances the clinical utility of urine chemistry testing.
Automation ; Blood Glucose ; Calcium ; Chemistry* ; Creatinine ; Diagnosis ; Limit of Detection ; Magnesium ; Nitrogen ; Urea ; Uric Acid

BACKGROUND: Point-of-care testing (POCT) is designed to be used near the site where the clinical care is being delivered. The demand for POCT in the medical field is expanding significantly, given that rapid results can eventually lead to early diagnosis and immediate clinical management of diseases. Therefore, the aim of this study was to evaluate the performance of the i-STAT POC analyser (Abbott Diagnostics, USA) for testing 8 chemical analytes (viz., sodium, potassium, chloride, total carbon dioxide, blood urea nitrogen, creatinine, glucose, and ionised calcium) and 2 hematological analytes (hematocrit [HCT], hemoglobin [Hb]). METHODS: The precision and linearity of the 10 analytes were measured according to Clinical and Laboratory Standards Institute (CLSI) EP15-A3 and EP6-A guidelines. Comparisons with a central laboratory hematology analyser, Coulter LH 780 (Beckman Coulter Inc., USA), and a chemical analyser, UniCel DxC 880i (Beckman Coulter Inc.), were performed using 85 patient samples according to CLSI EP9-A3. RESULTS: The coefficient of variation values for the within-run precision and total precision at 3 levels of all analytes were within 5%, except those for low level creatinine. In the aspect of linearity, the correlation coefficient values of all analytes were over 0.975 in the clinically important concentration range. A very high correlation was observed in glucose, blood urea nitrogen and creatinine (R>0.975), high correlation was observed in sodium, potassium, Hct and Hb (R>0.9), and relatively good correlation was observed in chloride and total carbon dioxide (R>0.7) compared to the central laboratory analysers. CONCLUSIONS: i-STAT showed relatively high precision and linearity, and comparable data to that of routine hematology and chemistry analysers. This device was concluded to have potential for providing faster results and relatively acceptable values to clinicians in need of immediate results.
Blood Glucose ; Blood Urea Nitrogen ; Carbon Dioxide ; Chemistry ; Creatinine ; Early Diagnosis ; Glucose ; Hematology ; Humans ; Nitrogen ; Point-of-Care Systems* ; Point-of-Care Testing ; Potassium ; Sodium ; Urea

The Clinical Chemistry Subcommittee of the Korean Association for External Quality Assessment Service conducted external quality assessments in 2015. This included general chemistry and blood gas measurements as part of a scheme of six trials, comprising three samples each. All control materials were included at the same time. The overall response rates were 94.4% for general chemistry and 92.2% for blood gas. The parameters tested included sodium, potassium, chloride, blood urea nitrogen, glucose, calcium, phosphorus, uric acid, creatinine, bilirubin, total protein, albumin, total cholesterol, triglyceride, aspartate aminotransferase (AST), alanine transaminase (ALT), alkaline phosphatase, lactate dehydrogenase, gamma glutamyl transferase, high-density lipoprotein (HDL) cholesterol, and low-density lipoprotein (LDL) cholesterol for general chemistry, and pH, partial pressure of carbon dioxide (pCO₂), and partial pressure of oxygen (pO₂) for blood gas assessments. Two types of reports were generated: a method summary, including mean, standard deviation, and coefficient of variation for each test method; and a result summary of each participating laboratory, including mean, standard deviation, number of peer groups, and standard deviation index and variance index scores of each laboratory. The overall quality performance in 2015 was similar to that of previous years, and showed lower interlaboratory variation than that in 2014. The requisite continual improvement in clinical chemistry testing quality can be achieved through participation in similar proficiency testing programs.
Alanine Transaminase ; Alkaline Phosphatase ; Aspartate Aminotransferases ; Bilirubin ; Blood Urea Nitrogen ; Calcium ; Carbon Dioxide ; Chemistry ; Chemistry, Clinical* ; Cholesterol ; Clinical Chemistry Tests ; Creatinine ; Glucose ; Hydrogen-Ion Concentration ; Korea* ; L-Lactate Dehydrogenase ; Lipoproteins ; Methods ; Oxygen ; Partial Pressure ; Peer Group ; Phosphorus ; Potassium ; Sodium ; Transferases ; Triglycerides ; Uric Acid

Nonhuman primates are increasingly used in biomedical research since they are highly homologous to humans compared to other rodent animals. However, there is limited reliable reference data of the clinical pathology parameters in cynomolgus monkeys, and in particular, only some coagulation and urinalysis parameters have been reported. Here, we reported the reference data of clinical chemical, hematological, blood coagulation, and urinalysis parameters in cynomolgus monkeys. The role of sex differences was analyzed and several parameters (including hematocrit, hemoglobin, red blood cell, blood urea nitrogen, total bilirubin, alkaline phosphatase, creatinine kinase, gamma-glutamyl tranferase, and lactate dehydrogenase) significantly differed between male and female subjects. In addition, compared to previous study results, lactate dehydrogenase, creatinine kinase, and aspartate aminotransferase showed significant variation. Interstudy differences could be affected by several factors, including age, sex, geographic origin, presence/absence of anesthetics, fasting state, and the analytical methods used. Therefore, it is important to deliberate with the overall reference indices. In conclusion, the current study provides a comprehensive and updated reference data of the clinical pathology parameters in cynomolgus monkeys and provides improved assessment criteria for evaluating preclinical studies or biomedical research.
Alkaline Phosphatase ; Anesthetics ; Animals ; Aspartate Aminotransferases ; Bilirubin ; Blood Coagulation ; Blood Urea Nitrogen ; Chemistry, Clinical ; Creatinine ; Erythrocytes ; Fasting ; Female ; Hematocrit ; Hematology ; Humans ; L-Lactate Dehydrogenase ; Lactic Acid ; Macaca fascicularis* ; Male ; Pathology, Clinical* ; Phosphotransferases ; Primates ; Reference Values* ; Rodentia ; Sex Characteristics ; Urinalysis

OBJECTIVETo explore the molecular mechanism for a boy suspected with 3-methylcrotonyl-CoA carboxylase deficiency by neonatal screening.

METHODSPCR and Sanger sequencing were used to identify potential mutations of MCCC1 and MCCC2 genes. SIFT and Polyphen-2 software was used to predict the effect of variant on the protein function and conservation of the variant across various species. Human Splicing Finder and Swiss-PdbViewer4.1.0 were applied to analyze the possible mechanism of the variant.

RESULTSFor the proband, a compound heterozygous mutation was discovered in the MCCC1 gene, namely c.539G>T (p.G180V) and c.704_711del (p.A235Vfs*4), which were inherited from his father and mother, respectively. The two mutations have disrupted the protein conformation, which in turn may impact the function of MCC protein.

CONCLUSIONThe compound heterozygous mutations of the MCCC1 gene may contribute to the 3-methylcrotonyl-CoA carboxylase deficiency manifested by the patient.

Amino Acid Sequence ; Base Sequence ; Carbon-Carbon Ligases ; chemistry ; deficiency ; genetics ; DNA Mutational Analysis ; Heterozygote ; Humans ; Infant, Newborn ; Male ; Models, Molecular ; Mutation ; Neonatal Screening ; methods ; Protein Conformation ; Sequence Homology, Amino Acid ; Urea Cycle Disorders, Inborn ; diagnosis ; genetics

Ethyl carbamate (EC) as a potential carcinogen commonly exists in traditional fermented foods. It is important eliminate urea that is the precursors of EC in many fermented foods, including Chinese Rice wine. On the basis of achieving high-level overexpression of food-grade ethanol-resistant acid urease, we studied the hydrolysis of urea and EC with the recombinant acid urease. Recombinant acid urease showed degraded urea in both the simulated system with ethanol and Chinese Rice wine (60 mg/L of urea was completely degraded within 25 h), indicating that the recombinant enzyme is suitable for the elimination of urea in Chinese Rice wine. Although recombinant acid urease also has degradation catalytic activity on EC, no obvious degradation of EC was observed. Further investigation results showed that the Km value for urea and EC of the recombinant acid urease was 0.7147 mmol/L and 41.32 mmol/L, respectively. The results provided theoretical foundation for realizing simultaneous degradation of urea and EC.
Oryza ; Recombinant Proteins ; metabolism ; Urea ; chemistry ; Urease ; metabolism ; Urethane ; chemistry ; Wine ; analysis

The Clinical Chemistry subcommittee of The Korean Association of External Quality Assessment Service conducted external quality assessments in 2014. This included general chemistry and blood gas measurements as part of a scheme of six trials, comprising of three samples each. All control materials were sent at the same time. The overall response rates were 93.4% for general chemistry and 90.0% for blood gas. The parameters tested included sodium, potassium, chloride, blood urea nitrogen, glucose, calcium, phosphorus, uric acid, creatinine, bilirubin, total protein, albumin, total cholesterol, triglyceride, AST, ALT, alkaline phosphatase, lactate dehydrogenase, gamma glutamyl transferase, HDL cholesterol, and LDL cholesterol for general chemistry and pH, partial pressure of carbon dioxide, and partial pressure of oxygen for blood gas assessment. Two types of reports were generated, namely, a method summary report including mean, standard deviation, and coefficient of variation, for each test method, as well as a result summary report of each participating laboratory, including mean, standard deviation, number of peer groups, and standard deviation index and variance index scores of each laboratory. The overall quality performance in 2014 was similar to that of previous years and as compared to results from 2013, the inter-laboratory variation was lower. The requisite continual improvement in the quality of clinical chemistry testing can be achieved through participation in similar proficiency testing programs.
Alkaline Phosphatase ; Bilirubin ; Blood Urea Nitrogen ; Calcium ; Carbon Dioxide ; Chemistry ; Chemistry, Clinical* ; Cholesterol ; Cholesterol, HDL ; Cholesterol, LDL ; Clinical Chemistry Tests ; Creatinine ; Glucose ; Hydrogen-Ion Concentration ; Korea* ; L-Lactate Dehydrogenase ; Laboratory Proficiency Testing ; Oxygen ; Partial Pressure ; Peer Group ; Phosphorus ; Potassium ; Research Report ; Sodium ; Transferases ; Triglycerides ; Uric Acid

BACKGROUND/AIMS: The aims of this study were to investigate whether a broccoli sprout extract containing sulforaphane (BSES) inhibited the Helicobacter pylori infection density and exerted an antioxidative effect on gastric mucosal damage. METHODS: The enrolled subjects were randomized in a double-blinded manner into three groups. Finally, 33 H. pylori (+) BSES treatment subjects (group A), 28 H. pylori (+) placebo subjects (group B), and 28 H. pylori (-) BSES treatment subjects (group C) were studied. H. pylori infection density was indirectly quantified by a 13C-urea breath test (UBT), and the ammonia concentration in gastric juice aspirates was measured through gastroscopic examination. Malondialdehyde (MDA), an oxidative damage biomarker, and reduced glutathione (GSH), an antioxidant biomarker, were measured in the gastric mucosa by an enzyme-linked immunosorbent assay. RESULTS: BSES treatment did not significantly affect the UBT values or ammonia concentration in group A (p=0.634 and p=0.505, respectively). BSES treatment did significantly reduce mucosal MDA concentrations in group A (p<0.05) and group C (p<0.001), whereas the gastric mucosal GSH concentrations did not differ before and after treatment in any of the groups. CONCLUSIONS: BSES did not inhibit the H. pylori infection density. However, BSES prevented lipid peroxidation in the gastric mucosa and may play a cytoprotective role in H. pylori-induced gastritis.
Adult ; Ammonia/metabolism ; Antioxidants/*pharmacology ; Biomarkers/analysis ; Brassica/*chemistry ; Breath Tests ; Double-Blind Method ; Enzyme-Linked Immunosorbent Assay ; Female ; Gastric Juice/enzymology ; Gastric Mucosa/*drug effects/metabolism ; Glutathione/analysis ; Helicobacter Infections/*drug therapy ; *Helicobacter pylori ; Humans ; Isothiocyanates/*pharmacology ; Lipid Peroxidation/*drug effects ; Male ; Malondialdehyde/analysis ; Middle Aged ; Plant Extracts/chemistry/*pharmacology ; Urea