Development of a simple prediction model for adrenal crisis diagnosis.

To develop a prediction model for adrenal crisis (AC) diagnosis among individuals with adrenal insufficiency that relies on the values of routinely measured clinical parameters, for application in standard clinical practice. We retrospectively analysed data from five referral centres in Japan. Multivariate binary logistic regression was used to identify independent predictors of AC, and receiver operating characteristic curve analysis was used to determine their optimal cut-off points. The analysis included data from 54 patients with 90 AC events. Logistic regression revealed that serum sodium and C-reactive protein (CRP) levels were independent predictors of AC. Serum sodium levels < 137 mEq/L had a sensitivity of 71.1% and specificity of 95.6%. CRP levels > 1.3 mg/dL had a sensitivity of 84.4% and specificity of 94.9%. In combination, serum sodium levels < 137 mEq/L or CRP levels > 1.3 mg/dL for AC diagnosis had sensitivity and specificity values of 97.8% and 94.4%, respectively. The combined use of serum sodium and CRP levels had high sensitivity and specificity, and can be used for AC screening in standard clinical practice. The model can assist in identifying AC among high-risk individuals. A larger prospective study is needed to validate these results.

Proposed cut-off value of CA19-9 for detecting pancreatic cancer in patients with diabetes: a case-control study.

Pancreatic cancer is a highly lethal malignancy. CA19-9 is a well-known marker for diagnosis of pancreatic cancer, but the serum CA19-9 level is reported to be elevated in patients with poorly controlled diabetes. This study evaluated the sensitivity, specificity, and cut-off value of serum CA19-9 for detection of pancreatic cancer in patients with diabetes. A case-control study of 236 patients was performed. The case group was selected from diabetic patients with pancreatic cancer, while one control was selected for each case from among diabetic patients without pancreatic cancer during the same period. The case group (n = 118) and the control group (n = 118) were matched for age, sex, and pancreatic cancer risk factors. Receiver operating characteristic (ROC) curves were plotted to determine the serum CA19-9 level that predicted pancreatic cancer. Then the sensitivity and specificity of CA19-9 were calculated for the threshold value. There were no significant differences of age, sex, BMI, smoking, alcohol intake, and HbA1c between the case and control groups. According to ROC analysis, a serum CA19-9 level of 75 U/mL had the maximum sensitivity and specificity for separating diabetic patients with or without pancreatic cancer. Using this cut-off value, the sensitivity and specificity of CA19-9 for pancreatic cancer was 69.5% and 98.2%, respectively, while the area under the ROC curve was 0.875 [95%CI: 0.826-0.924]. We propose that a serum CA19-9 level of 75 U/mL should be used as the cut-off value when screening patients with diabetes for pancreatic cancer.

Efficacy and safety of switching from basal insulin to once-daily insulin degludec/insulin aspart in Japanese patients with inadequately controlled type 2 diabetes: A 4-week, randomized, open-label, treat-to-target study.

AIMS/INTRODUCTION: A prospective, 4-week, single-center, randomized, open-label, parallel-group, treat-to-target study was carried out to develop an algorithm for safe and effective switching from basal insulin to once-daily insulin degludec/insulin aspart (IDegAsp) in patients with inadequately controlled type 2 diabetes. MATERIALS AND METHODS: Patients were randomly assigned to continue their current basal insulin therapy (n = 10) or to switch to IDegAsp on a 1:1 unit basis (n = 10). The insulin dose could be titrated once weekly, targeting a self-measured blood glucose of 80-100 mg/dL before breakfast. A mixed meal test was carried out at baseline and after 4 weeks. RESULTS: After 4 weeks, the mean daily dose of insulin was similarly increased by 60% in both groups, and there was a significant decrease of mean plasma glucose and glucose area under the glucose concentration vs time curve for 2 h in the meal test. The mean estimated treatment difference (IDegAsp group - basal insulin group) of the mean plasma glucose level was -28 mg/dL (95% confidence interval -47 to -8, P = 0.008) after 4 weeks and that of the area under the glucose concentration vs time curve for 2 h was -2,800 mg/min/dL (95% confidence interval -5,300 to -350, P = 0.028), confirming the superiority of IDegAsp to basal insulin. In the IDegAsp group, the 2-h postprandial plasma glucose level was significantly decreased to the fasting plasma glucose range. There were no confirmed hypoglycemic episodes in either group during the 4-week study period. CONCLUSIONS: After switching from basal insulin, the IDegAsp dose can be uptitrated by 60% based on fasting plasma glucose data. However, monitoring of postprandial glucose should be considered before further uptitration of IDegAsp.

Efficacy and safety of thrice-weekly insulin degludec in elderly patients with type 2 diabetes assessed by continuous glucose monitoring.

As life expectancy becomes longer in Japan, there has been an increase of elderly patients with type 2 diabetes who need insulin therapy but cannot perform self-injection due to dementia or other conditions. Therefore, the aim of this study was to investigate the efficacy and safety of thrice-weekly insulin degludec therapy in elderly patients with poorly controlled diabetes. The subjects were 22 hospitalized elderly Japanese patients with type 2 diabetes who had difficulty with self-injection. After becoming stable on once-daily insulin degludec treatment, they were assigned to continue once-daily injection (OD group) or were switched to thrice-weekly injection (TW group) for one week. In the TW group, insulin degludec (IDeg) was injected at twice the OD dose before lunch on Monday, Wednesday, and Friday. Glycemic control was assessed by continuous glucose monitoring (CGM) over 7 days. The mean 7-day glucose level (131±25 mg/dL with OD vs. 152±30 mg/dL with TW, p=0.11) and the mean 7-day standard deviation (32±10 mg/dL with OD vs. 36±14 mg/dL with TW, p=0.45) did not differ significantly between the two groups. The percent duration of glucose <70 mg/dL (2.4±3.1% with OD vs. 1.3±2.5% with TW, p=0.39) and glucose >200 mg/dL (7.2±12.1% with OD vs. 15.6±18.0% with TW, p=0.22) over 7 days also showed no significant differences between the two groups. In conclusion, thrice-weekly IDeg provided by a visiting nurse could be a practical option for elderly diabetic patients who have difficulty performing self-injection of insulin.

The role of the endocrine system in feeding-induced tissue-specific circadian entrainment.

The circadian clock is entrained to environmental cycles by external cue-mediated phase adjustment. Although the light input pathway has been well defined, the mechanism of feeding-induced phase resetting remains unclear. The tissue-specific sensitivity of peripheral entrainment to feeding suggests the involvement of multiple pathways, including humoral and neuronal signals. Previous in vitro studies with cultured cells indicate that endocrine factors may function as entrainment cues for peripheral clocks. However, blood-borne factors that are well characterized in actual feeding-induced resetting have yet to be identified. Here, we report that insulin may be involved in feeding-induced tissue-type-dependent entrainment in vivo. In ex vivo culture experiments, insulin-induced phase shift in peripheral clocks was dependent on tissue type, which was consistent with tissue-specific insulin sensitivity, and peripheral entrainment in insulin-sensitive tissues involved PI3K- and MAPK-mediated signaling pathways. These results suggest that insulin may be an immediate early factor in feeding-mediated tissue-specific entrainment.

Efficacy of low-dose rosuvastatin in patients with type 2 diabetes and hypo high-density lipoprotein cholesterolaemia.

OBJECTIVE: To analyse the efficacy of low-dose rosuvastatin for treating hypo high-density lipoprotein (HDL) cholesterolaemia in patients with type 2 diabetes and dyslipidaemia. METHODS: Patients with HDL-cholesterol (C) < 40 mg/dl and triglycerides (TG) < 400 mg/dl who were receiving treatment with lipid-lowering drugs other than rosuvastatin (or previously untreated with lipid-lowering drugs) and with low-density lipoprotein [LDL]-C ≥ 120 mg/dl were included. Patients were treated with 2.5 or 5 mg rosuvastatin orally, once daily, to achieve the target LDL-C level specified in Japanese guidelines. Changes in total cholesterol, HDL-C, TG, LDL-C, LDL-C/HDL-C and non-HDL-C at 3 and 6 months were prospectively analysed. Safety was evaluated by examining changes in hepatorenal function, glucose metabolism and creatine kinase. RESULTS: Out of 49 patients, all lipid parameters other than TG were significantly improved at 3 and 6 months. At 3 months, 83.3% of patients had achieved the target LDL-C level. Among nonlipid parameters, no changes were observed except for estimated glomerular filtration rate, which was improved by + 5.2% and + 9.6% at 3 and 6 months, respectively. CONCLUSIONS: Low-dose rosuvastatin was effective in improving hypo-HDL cholesterolaemia and may have renoprotective effects.

Effect of insulin glargine on endogenous insulin secretion and beta-cell function in Japanese type 2 diabetic patients using oral antidiabetic drugs.

The aim of the present study was to evaluate the effect of insulin glargine (Gla) (as part of basal-supported oral therapy) on endogenous insulin secretion and beta-cell function in type 2 diabetic patients. In 33 insulin-naive patients showing poor glycemic control on treatment with sulfonylurea (SU)-based OADs without DPP4 inhibitors, once-daily injection of Gla was added without changing OADs, and the dose of Gla was titrated to attain a fasting plasma glucose (FPG) <110 mg/dL over 24 weeks. Morning meal tests were done at baseline, 12 weeks and 24 weeks. FPG and 2-hour plasma glucose (2HPG) and serum C-peptide (FCPR and 2HCPR) were measured 3 times, while serum intact proinsulin (FPI and 2HPI) was measured at baseline and 24 weeks. Levels of FPG, FCPR, 2HPG, and HbA1c were significantly reduced from baseline at 24 weeks (176±52 to 117±27 mg/dL, p<0.01; 2.0±0.9 to 1.6±1.0 ng/mL, p<0.01; 257±53 to 202±27 mg/dL, p<0.01; and 8.4±0.9 to 7.3±0.6%, p<0.01, Mean±SD), but 2HCPR was unchanged. The patients were divided into two groups depending on whether FPG at 24 weeks was <110 mg/dL or not: attained group (n=15) and not attained group (n=18). The dose of Gla did not differ between the two groups, but the 2HPI/2HCPR ratio at 24 weeks showed a significant decrease from baseline in the attained group. Supplementation with Gla improved glycemic control and maintained intrinsic basal insulin secretion, without changing 2-hour postprandial secretion. Achieving good glycemic control with an FPG<110 mg/dL by adding Gla decreased the 2HPI/2HCPR ratio at 24 weeks.

Contamination of poultry products with Listeria monocytogenes at poultry processing plants.

This study aimed to confirm that poultry products packed at poultry processing plants have already been contaminated with Listeria monocytogenes and that poultry products contaminated with L. monocytogenes are derived from broiler flocks infected with L. monocytogenes. L. monocytogenes was isolated from 16.8% (58/345) of chicken breast products and 2.3% (8/345) of chicken liver products. In contrast, L. monocytogenes was isolated from the pooled cecal content sample from only 1 (4%) of 25 flocks and was never isolated from any pooled dropping samples collected from 25 farms. The results of our study indicate that cecal content does not seem to be an important source of L. monocytogenes in poultry products.

Prevalence and antimicrobial susceptibility of foodborne bacteria in wild boars (Sus scrofa) and wild deer (Cervus nippon) in Japan.

This study aimed to evaluate the role of wild boars and deer as reservoirs of foodborne bacteria. We investigated the prevalence and antimicrobial susceptibility of Campylobacter spp., Salmonella spp., Shiga toxin-producing Escherichia coli (STEC) O157 and O26, and Listeria monocytogenes isolated from wild boars and deer in Japan, from July through December 2010. Campylobacter spp. and Salmonella spp. were isolated from 43.8% (95% confidence interval [CI]: 35.0-52.6) and 7.4% (95% CI: 2.8-12.1) of rectal content samples of wild boars, respectively, but not from wild deer. The most common Campylobacter species was C. lanienae and C. hyointestinalis. The nine Salmonella serovars isolated were S. enterica subsp. enterica serovar Agona (three isolates), S. Narashino (two), S. Enteritidis (one), S. Havana (one), S. Infantis (one), and S. Thompson (one). Five (16%) and 6 (29%) isolates of C. lanienae and C. hyointestinalis, respectively, were resistant to enrofloxacin. STEC O157 and O26 and L. monocytogenes were isolated from 2.3% (95% CI: 0-5.0), 0.8% (95% CI: 0-2.3), and 6.1% (95% CI: 1.7-10.5) of the rectal content samples of wild deer, respectively, but not from wild boars. This first nationwide survey of the prevalence of foodborne bacteria in wild boars and wild deer in Japan suggests that consumption of meat from these animals is associated with the risk of causing infection with these bacteria in humans. Moreover, these animals are potential vehicles for distribution of antimicrobial-resistant bacteria into their habitat. The prevalence and antimicrobial susceptibility of such foodborne bacteria in these wild animals should be monitored periodically.

Comparison of the prevalence of shiga toxin-producing Escherichia coli strains O157 and O26 between beef and dairy cattle in Japan.

With the aim of comparing the prevalence of Shiga toxin-producing Escherichia coli (STEC) O157 and O26 between beef and dairy cattle, we collected rectal content samples from 250 beef cattle on 25 beef farms and 250 dairy cows on 25 dairy farms from July through September 2011. STEC O157 was isolated from 16 beef cattle on 7 beef farms, while no STEC O157 was isolated from any dairy farms. This result suggests that the prevalence of STEC O157 is higher in beef cattle than in dairy cattle. STEC O26 was isolated from 1 animal each from beef and dairy cattle herds, and therefore, it was not possible to compare statistically the prevalence of STEC O26 in beef and dairy cattle.

Prevalence of Campylobacter spp., Salmonella spp., Listeria monocytogenes, and hepatitis E virus in swine livers collected at an abattoir.

We investigated the prevalence of Campylobacter spp., Salmonella spp., Listeria monocytogenes, and hepatitis E virus (HEV) in swine liver. We collected swine livers from 110 pigs at an abattoir from September 2011 to March 2012 [corrected] . Pathogens were detected in the liver samples of 19 (17.3%) pigs. Campylobacter spp. were isolated from the liver samples of 14 (12.7%) pigs. In 10 of the 14 Campylobacter-positive pigs, bacteria were present in the internal regions of the liver. Salmonella spp. and L. monocytogenes were detected in the liver samples of 5 (4.5%) pigs and 1 (1%) pig, respectively. No HEV was detected in the swine liver samples tested. Regarding antimicrobial resistance in Campylobacter and Salmonella isolates, all isolates, except 1 Campylobacter jejuni isolate, were resistant to 1 or more antimicrobial agent. Campylobacter spp. resistant to erythromycin and/or enrofloxacin were isolated from the liver samples of 9 (8%) pigs. These results suggest that the consuming swine liver without proper heat treatment may increase the risk of foodborne illnesses.

Prevalence and antimicrobial resistance of Campylobacter isolates from beef cattle and pigs in Japan.

Rectal contents of beef cattle and pigs were collected between October 2010 and February 2011 in Japan. Campylobacter jejuni were isolated from 36.0% (90/250) of beef cattle from 88.0% (22/25) of beef farms. C. coli were isolated from 3.6% (9/250) of beef cattle from 16.0% (4/25) of beef farms and from 42.4% (106/250) of pigs from all 25 pig farms. As to enrofloxacin, 40.0% (36/90) of C. jejuni isolates and 66.7% (6/9) of C. coli isolates from beef cattle and 44.3% (47/106) of C. coli isolates from pigs were resistant. Additionally, 15.1% (16/106) of C. coli isolates from pigs were resistant to erythromycin and enrofloxacin. The high prevalence of Campylobacter carriers and significant antimicrobial resistance of the isolates were found.

Prevalence and characterization of foodborne pathogens in dairy cattle in the eastern part of Japan.

To investigate the prevalence and characterization of foodborne pathogens [Campylobacter spp., Shiga toxin-producing Escherichia coli (STEC), Listeria monocytogenes and Salmonella spp.] in dairy cows, rectal content grab samples were collected from 250 dairy cows reared on 25 dairy farms in eastern Japan from December 2010 through February 2011. Campylobacter jejuni was isolated from 106 (42%) cows on 23 (92%) farms, STEC O157 from three cows on one farm, L. monocytogenes from three cows on another three farms and Salmonella enterica subsp. enterica serovar Typhimurium from eight cows on another farm. STEC O26 was not isolated from any of the dairy farms investigated. The results suggest that C. jejuni is widespread in dairy farms in eastern Japan.

Antimicrobial resistance in Shiga toxin-producing Escherichia coli O157 and O26 isolates from beef cattle.

This study was conducted to determine the prevalence of antimicrobial resistance in Shiga toxin-producing Escherichia coli (STEC) O157 (n = 241) and O26 (n = 11) isolated from beef cattle and to characterize their antimicrobial resistance profiles. Resistance to dihydrostreptomycin was detected most frequently (STEC O157, 9.5%; STEC O26, 54.5%), followed by resistance to oxytetracycline (7.9%; 45.5%) and ampicillin (5.4%; 36.4%). Resistance to one or more antimicrobial agents was detected in 13.3% (32/241) of the STEC O157 isolates and 54.5% (6/11) of the STEC O26 isolates. The antimicrobial resistance rate in the STEC O26 isolates was significantly higher than that in the STEC O157 isolates (P = 0.002, Fisher's exact test). The antimicrobial resistance rate in the STEC O157 isolates possessing both stx(1) and stx(2) genes was 26.3% (15/57), while that in the isolates possessing stx(2c) gene alone was 3.9% (3/77). These findings suggest that the antimicrobial resistance in STEC O157 is associated with serogroups and the Shiga toxin genotype.

[Establishment of a cut-off value for the diagnosis of acute myocardial infarction using a new CK-MB activity measurement reagent containing anti-MtCK antibody].

CK-MB is an important marker for the diagnosis of acute myocardial infarction (AMI). Since mitochondrial CK (MtCK) is universally present in the blood of healthy individuals, it is known to positively affect the measurement of CK-MB using the immunoinhibition method, causing false-positive results. We performed basic evaluation of ACCURAS AUTO CK-MB MtO, a new reagent containing anti-MtCK antibody that inhibits MtCK activity, and attempted to calculate a cut-off CK-MB level to diagnose AMI. The measurement was performed in samples submitted to the Clinical Laboratory of our center for the measurement of CK-MB. This method was confirmed to have satisfactory basic attributes concerning the reproducibility, linearity, lower detection limit, and effects of interfering substances. When 2886 samples were examined using this and conventional methods, the results of the two methods were correlated in some but not in others. In the samples that showed no correlation, MtCK was demonstrated by isozyme analysis using electrophoresis. The AUC calculated from the ROC curve in AMI patients was 0.912 with this method and 0.861 with the conventional method. The sensitivity and specificity of the new method were higher than those of the conventional method. The cut-off value determined by ROC analysis was 7.7 U/l using the new method and 13.6 U/l using the conventional method, causing an increase in false-positive results compared with the cut off value of 25 U/l widely used for the conventional method to date. However, the cut-off value for the new method that yielded a specificity comparable to 99.1%, which is the specificity of the conventional method using a cut-off value of 25 U/l, was 12 U/l. With a cut-off value of 12 U/l, the sensitivity was improved compared with that employing the conventional method, and both the sensitivity and specificity became comparable to those of the CK-MB mass method. This method is very useful for the accurate measurement of CK-MB activity.

Effects of caloric intake on intestinal mucosal morphology and immune cells in rats treated with 5-Fluorouracil.

Anticancer drugs have been reported to damage the intestinal mucosa. We evaluated the effects of caloric intake on the mucosal morphology and immune cells in rats treated with 5-fluorouracil (5-FU). Rats were received a liquid diet plus 5-FU treatment for 8 days as follows: Low calorie group (25 kcal/day with 5-FU), Normal calorie group (50 kcal/day with 5-FU), and Control group (50 kcal/day with saline). The mucosal morphology, cell numbers and phenotypes of spleen and intraepithelial lymphocytes (IEL) were assessed. As compared with the control group, the villus heights were significantly lower in the Low calorie group, but not significantly lower in the Normal calorie group. The total cell yield from the spleen, CD4+ and CD8+ T cells decreased in the Low and Normal calorie group, but these changes were less pronounced in Normal calorie group. The total cell yield from the IEL also decreased in the Low calorie group, but not in the Normal calorie group. Our study demonstrated that sufficient caloric intake attenuated the damages in intestinal morphology and in the immune cell numbers. Clinically, nutritional support would be expected to be one approach to reducing the risk of bacterial translocation or infection induced by chemotherapy.

Prevalence of fowl glioma-inducing virus in chickens of zoological gardens in Japan and nucleotide variation in the env gene.

Fowl glioma-inducing virus (FGV), which belongs to subgroup A of avian leukosis virus (ALV), is tumorigenic in the nervous system. In a zoological garden in Japan, approximately 40% of chickens, including Japanese fowls, were infected with FGV. Because this zoological garden plays a role as a major supplier of Japanese fowl for other zoological gardens, FGV infection is suspected to have spread among ornamental chickens. In this study, the prevalence of the disease was examined in a total of 129 chickens in three other zoological gardens by nested polymerase chain reaction (PCR), reverse transcription nested PCR and enzyme-linked immunosorbent assay. Twenty-six to 56 percent of the fowls in each of the examined gardens were positive by nested PCR. The phylogenetic analysis revealed that the 3' untranslated region, including the specific sequence of FGV, of the 14 isolated ALVs showed high sequence identity and a close relationship with FGV. In addition, the env gene of the isolates frequently showed mutations and deletions of nucleotides. These results suggest that FGV is prevalent among ornamental chickens kept in zoological gardens in Japan.

Changes in lymphocyte phenotypes and cytokine production by surgical stress in a rat small intestinal resection model.

Small intestinal resection rats are used widely as a malabsorption model, but the immunological changes are unclear. We examined the changes in systemic and mucosal immune status after a small intestinal resection in rats with a controlled nutritional status. Rats had 60% of their small intestine removed. At 5 days after the surgery, spleen cells and intraepithelial lymphocytes (IEL) were isolated. The phenotypes of spleen cells and IEL, the production patterns of Th1 and Th2 cytokines, and the proinflammatory cytokine levels in the plasma were measured. CD4+ T cells in the blood and spleen were significantly decreased in the Resection group (p<0.05). In contrast, IEL subpopulations were not different between the two groups. Interferon-gamma production from the spleen cells was significantly decreased in the Resection group (p<0.05). Interleukin (IL)-4 production was not different between the two groups. Plasma IL-6 concentrations were significantly elevated in the Resection group 6 h after surgery (p<0.05). In conclusions, small intestinal resection in rats suppressed systemic immunity, and this model is useful as a surgical stress model.

Characterization of a haemolytic factor from Candida albicans.

The culture supernatant of Candida albicans promoted the disruption of human red blood cells (RBCs). The haemolytic activity was detected in a sugar-rich fraction (about 200 kDa) from Sephacryl S-100 chromatography. As the haemolytic activity was adsorbed by concanavalin A-Sepharose, the haemolytic factor may be a mannoprotein. The activity was inactivated by periodate oxidation, indicating that the sugar moiety of the mannoprotein played an important role in the haemolysis. The structure of the sugar moiety of the mannoprotein was identified as a cell-wall mannan by 1H-NMR analysis, and purified C. albicans mannan promoted the disruption of RBCs. The binding of mannan to RBCs was demonstrated by flow cytometric analysis and was inhibited by the addition of band 3 protein inhibitor, 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS). The haemolysis caused by mannan was inhibited by DIDS, SITS (4-acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic acid) and bis(sulfosuccinimidyl) suberate, but not by pyridoxal 5-phosphate. These results indicated that a mannoprotein released from C. albicans bound to the band 3 protein on RBCs, thereby promoting their disruption.