Angiotensin-converting enzyme inhibition curbs tyrosine nitration of mitochondrial proteins in the renal cortex during the early stage of diabetes mellitus in rats.

Experiments were performed to evaluate the hypothesis that ACE (angiotensin-converting enzyme) inhibition (enalapril) suppresses 3-NT (3-nitrotyrosine) production in the renal cortex during the early stage of Type 1 DM (diabetes mellitus) in the rat. Enalapril was administered chronically for 2 weeks to subsets of STZ (streptozotocin)-induced DM and vehicle-treated sham rats. O(2)(-) (superoxide anion) and NO(x) (nitrate+nitrite) levels were measured in the media bathing renal cortical slices after 90 min incubation in vitro. SOD (superoxide dismutase) activity and 3-NT content were measured in the renal cortex homogenate. Renal cortical nitrated protein was identified by proteomic analysis. Renal cortical production of O(2)(-) and 3-NT was increased in DM rats; however, enalapril suppressed these changes. DM rats also exhibited elevated renal cortical NO(x) production and SOD activity, and these changes were magnified by enalapril treatment. 2-DE (two-dimensional gel electrophoresis)-based Western blotting revealed more than 20 spots with positive 3-NT immunoreactivity in the renal cortex of DM rats. Enalapril treatment blunted the DM-induced increase in tyrosine nitration of three proteins ACO2, GDH1 and MMSDH (aconitase 2, glutamate dehydrogenase 1 and methylmalonate-semialdehyde dehydrogenase), each of which resides in mitochondria. These data are consistent with enalapril preventing DM-induced tyrosine nitration of mitochondrial proteins by a mechanism involving suppression of oxidant production and enhancement of antioxidant capacity, including SOD activation.

[Approach to the changes of HbA1c notation in health checkups].

The Committee on the Standardization of Diabetes Mellitus-Related Laboratory Testing of the Japan Diabetes Society (JSD) released "International clinical harmonization of hemoglobin A1c in Japan: From JDS to NGSP values". In the document, the Japanese Ministry of Health, Labor, and Welfare, related academic societies, manufacturers and others, and the council meeting of JDS finally decided to use NGSP values in clinical practice from April 1, 2012. They also decided to use the JDS value as usual at specific health checkups and in specific health guidance for one year since the conversion of a great number of digital data needs a lot of time and is costly, and the conversion influences assessment systems markedly. As soon as the document was released, our laboratory (commercial laboratory) noticed that clients at clinics, hospitals and health checkup institutions were aware of the change (or no change) of the HbA1c notation. Subsequently, we set up analysis equipment to output the NGSP value and altered the settings of the laboratory information system to convert the JDS value from the NGSP value. In fact, the health checkup institutions took only the JDS value and the clinics and hospitals took the NGSP and JDS values together. In October 2012, JDS decided that only the NGSP value should be used at specific health checkups and in specific health guidance from April 1, 2013. In conclusion, the performance of international harmonization of HbA1c in Japan was considered to be of great significance.

Effects of supplemental ß-carotene on colostral immunoglobulin and plasma ß-carotene and immunoglobulin in Japanese Black cows.

Data from 26 Japanese Black cows were collected to clarify the effects of supplemental ß-carotene on colostral immunoglobulin (Ig) and plasma ß-carotene and Ig in the cows. Cows were assigned to control or ß-carotene groups from 21 days before the expected calving date to 60 days after parturition. Supplemental ß-carotene was provided at 500 mg/day in the ß-carotene group. Supplemental ß-carotene drastically increased plasma ß-carotene concentrations in the cows from parturition to 60 days after parturition, and plasma ß-carotene concentrations in the control and ß-carotene groups at parturition were 202 and 452 µg/dl, respectively. Supplemental ß-carotene had no effects on plasma IgG1 , IgA or IgM concentrations at parturition. Supplemental ß-carotene increased colostral IgG1 concentrations in the cows, but colostral ß-carotene, IgA and IgM concentrations were not affected by supplemental ß-carotene. These results indicate that supplemental ß-carotene is effective to enhance colostral IgG1 concentrations and plasma ß-carotene concentrations in Japanese Black cows.

Impact of angiotensin-converting enzyme inhibition on renal cortical nitrotyrosine content during increased extracellular glucose concentration.

OBJECTIVES: Experiments evaluated the hypothesis that angiotensin-converting enzyme (ACE) inhibition suppresses hyperglycemia-induced nitrotyrosine (NT) production in the renal cortex. DESIGN AND METHODS: Rats were untreated (UNTR, n = 6) or received the ACE inhibitor enalapril (20 mg/kg/day; ENAL, n = 6) for 2 weeks. Renal cortical slices were incubated for 90 min in media containing 5 (normal) or 20 mmol/L (high) glucose. Superoxide anion (O2*-) and nitrate + nitrite (NO(X)) levels were measured in the media. Superoxide dismutase (SOD) activity and NT content were measured in the tissue homogenate. RESULTS: In the UNTR group, high glucose increased O2*- and NO(X) production by the renal cortex (P < 0.05 vs. normal glucose). Likewise, NT content and SOD activity of the renal cortex augmented (P < 0.05 vs. normal glucose). In the ENAL group, O2*- production and NT content were glucose-insensitive, but high glucose exerted an exaggerated impact on NO(X) production and SOD activity (P < 0.01 vs. UNTR in high glucose). CONCLUSION: Accelerated NT content in the renal cortex during high-glucose conditions was prevented by ACE inhibitor treatment. It was suggested that, apart from its anti-hypertensive effect, the mechanism of suppressed NT degradation in the renal cortex by the ACE inhibitor enhances both O2*- degradation per se and antioxidative effects including SOD activation.

[Age- and sex-specific reference intervals for 10 health examination items: mega-data from a Japanese Health Service Association].

OBJECTIVE: To determine age- and sex-specific reference intervals for 10 health examination items in Japanese adults. METHODS: Health examination data were accumulated from 24 different prefectural health service associations affiliated with the Japan Association of Health Service. Those who were non-smokers, drank less than 7 days/week, and had a body mass index of 18.5-24.9kg/m2 were sampled as a reference population (n = 737,538; 224,947 men and 512,591 women). After classified by age and sex, reference intervals for 10 health examination items (systolic blood pressure, diastolic blood pressure, total cholesterol, triglyceride, glucose, uric acid, AST, ALT, gamma-GT, and hemoglobin) were estimated using the parametric and nonparametric methods. RESULTS: In every item except for hemoglobin, men had higher reference intervals than women. Systolic blood pressure, total cholesterol, and glucose showed an upward trend in values with increasing age. Hemoglobin showed a downward trend in values with increasing age. Triglyceride, ALT, and gamma-GT reached a peak in middle age. Overall, parametric estimates showed narrower reference intervals than non-parametric estimates. CONCLUSION: Reference intervals vary with age and sex. Age- and sex-specific reference intervals may contribute to better assessment of health examination data.

High glucose augments arginase activity and nitric oxide production in the renal cortex.

To clarify the interaction between arginase and nitric oxide (NO) production in the kidney with normal and high glucose levels, renal cortical slices from male Sprague-Dawley rats were incubated in Hank's solution containing various concentrations of L-norvaline (Nval; an arginase inhibitor), 500 U/mL superoxide dismutase, and either 5 mmol/L (normal) or 20 mmol/L (high) glucose (n = 5 per group). Incubation with Nval increased renal cortical NOX (nitrite + nitrate) production dose-dependently, indicating competition between arginase and NO synthase (NOS) for the substrate (L-arginine). In the basal condition without Nval, high glucose also increased NO(X) production to a rate 3 times that observed during incubation with normal glucose (P < .01). This effect of high glucose was not altered by Nval. Rather, the effects of high glucose and Nval were additive, indicating that the activity of NOS per se is enhanced by high glucose. Direct assay of arginase and NOS activities confirmed stimulation of both enzymes under the high glucose condition (P < .05, P < .01, v normal glucose, respectively). However, high glucose did not change the amount of L-arginine present in renal cortical slices. These data reveal that arginase competes with NOS for L-arginine in the renal cortex, and that high glucose increases the activity of both enzymes without affecting the amount of substrate. These results suggest that increased NOS activity, rather than altered substrate availability, may be the principal factor underlying increased NO synthesis in diabetic kidneys.

Thrombomodulin alfa in the treatment of infectious patients complicated by disseminated intravascular coagulation: subanalysis from the phase 3 trial.

To investigate treatment effects of thrombomodulin alfa (TM-α) in patients with disseminated intravascular coagulation (DIC) having infection as the underlying disease, retrospective subanalysis of a double-blind, randomized controlled phase 3 trial was conducted. In the phase 3 trial, 227 DIC patients (full-analysis set) having infection and/or hematologic malignancy as the underlying disease received either TM-α (0.06 mg·kg for 30 min once daily) or heparin (8 U·kg·h for 24 h) for 6 days using the double-dummy method. Among these patients, 147 patients with noninfectious comorbidity leading to severe thrombocytopenia (e.g., hematologic malignancy, or aplastic anemia) were excluded from the present analysis, and 80 patients with infectious disease and DIC were extracted and subjected to the present retrospective subanalysis. Disseminated intravascular coagulation resolution rates were determined using the DIC diagnostic criteria for critically ill patients at 7 days, and mortality rates were evaluated at 28 days. In the TM-α and heparin groups, DIC resolution rates were 67.5% (27/40) and 55.6% (20/36), and 28-day mortality rates were 21.4% (9/42) and 31.6% (12/38), respectively. Mortality rates of patients who recovered from DIC were 3.7% (1/27) in the TM-α group and 15% (3/20) in the heparin group. These results suggest TM-α may be valuable in the treatment of DIC associated with infection.

Commutability of National Institute of Standards and Technology standard reference material 1955 homocysteine and folate in frozen human serum for total folate with automated assays.

BACKGROUND: The aim of the present study was to evaluate standard reference material (SRM) 1955 commutability as a reference material for serum folate using automated methods. We also designed so as to reduce the intermethod variability present in different automated methods. METHODS: Using a microbiological assay related to the 'information value' of SRM 1955 as a comparison method, we investigated the possibility of standardization for the assay values of serum folate as measured by the automated methods (Access, Centaur and Elecsys). In the assay of 50 patient sera by these automated methods, we corrected observed values by the SRM 1955 and compared with comparison values. RESULTS: The observed values of SRM 1955 Levels I, II and III were within or outside (but near) a 95% prediction interval obtained from patient sera by the automated methods. The normalized residuals obtained from SRM 1955 were within ±3.0 (in SD units), which enabled us to conclude that the SRM 1955 had a physicochemical characterization similar to native serum. Twelve patients were assessed as hypofolataemia (<6.0 ng/mL) and 38 patients as normal (≥6.0 ng/mL). Before correction, folate levels in six of 12 patients were lower than 6.0 ng/mL, and those in seven of 38 patients were higher than 6.0 ng/mL with the automated methods. After correction, low levels were found in four of 12 patients, and normal levels were found in 33 of 38 patients. CONCLUSIONS: The use of SRM 1955 would help to reduce the intermethod variability present in different automated methods for serum folate measurement.

Virological significance of low-level hepatitis B virus infection in patients with hepatitis C virus associated liver disease.

The clinical and virological significance of low-level viremia by hepatitis B virus (HBV) in hepatitis C virus (HCV)-infected patients remains unclear. HBV-DNA and HCV-RNA were, therefore, quantitatively analyzed in livers and sera from co-infected patients. HBV-DNA and HCV-RNA were quantitated using real-time detection of polymerase chain reaction (RTD-PCR), based on Taq-Man chemistry, in 220 non-HCV-infected healthy volunteers and 93 HCV-infected patients without detectable HBsAg. Serum HBV-DNA was detected in 4 (1.8%) of 220 non-HCV-infected healthy volunteers and 32 (34.4%) of 93 HCV-infected patients without detectable HBsAg. HCV-infected patients displayed higher frequency of HBV infection than healthy volunteers (P < 0.0001). Hepatocellular carcinoma (HCC) was more frequent among co-infected patients than among HCV mono-infected patients (P < 0.001). However, quantities of HBV-DNA in sera from co-infected patients were very low (8-19,000 copies/ml). HBV-DNA was detected in liver tissue from co-infected patients at 2-20 copies per 100 hepatocytes, accounting for 1/1,000 to 1/10,000 of HBsAg positive patients. In livers of patients with HCC and HCV or HBV mono-infection, the viruses existed predominantly in non-cancerous tissue, with levels 10- to 1,000-fold and 1- to 100-fold higher than in cancerous tissue, respectively. In contrast, patients co-infected with HCV and HBV displayed decreased HBV levels in non-cancerous tissue, but no change in cancerous tissue. These results indicate that low-level HBV infection exists in HCV-infected patients. HCC was more common among HCV/HBV co-infected patients than among HCV mono-infected patients. HCV might initiate hepatocarcinogenesis, but does not necessarily determine progression to HCC.