Dietary Survey of Japanese Individuals with Type 2 Diabetes Mellitus on a Low-Carbohydrate Diet: An Observational Study.

The nutrient intake of persons with diabetes placed on a low-carbohydrate diet remains unclear. This study aimed to assess nutrient intake in persons with type 2 diabetes mellitus treated with a low-carbohydrate diet. The brief-type self-administered diet history questionnaire was used to collect the dietary information of 335 outpatients at Kitasato Institute Hospital, while their clinical characteristics were collected from their electronic medical records. The median age, HbA1c level, and body mass index of the participants were 68 (60-74) years, 49 (45-55) mmol/mol [6.7 (6.3-7.2)%], and 24.0 (21.8-26.7) kg/m2, respectively; median energy intake was 1457 (1153-1786) kcal/day; and protein-energy, fat-energy, and available carbohydrate-energy ratios were 18.6 (15.7-21.4)%E, 36.8 (31.6-43.2)%E, and 34.6 (26.0-42.4)%E, respectively. As the available carbohydrate-energy ratio decreased, the fat-energy ratio increased significantly. The total dietary fibre and salt intake were 7.1 (5.6-8.4) g/1000 kcal and 6.5 (5.6-7.5) g/1000 kcal, respectively. Japanese individuals with type 2 diabetes mellitus placed on a low-carbohydrate diet had a fat-to-energy ratio exceeding 30%, while the fat-energy ratio increased as the carbohydrate-energy ratio decreased.

Team-based Learning Impact: A Comparative Study of Student and Faculty Facilitators.

BACKGROUND: Team-based learning (TBL) refers to the application of an active-learning method that has gained popularity across all health-care disciplines. This study aimed to assess nutrition students' perceptions of the roles of student versus faculty facilitators. METHODS: Participants in the study included, 117 2nd-year nutrition students registered in the "Introduction to Medicine" course in the 2022 academic year at a Japanese university. The first TBL session was faculty-led, whereas three students served as facilitators in the second. Upon completion of the course, learners and student facilitators completed a questionnaire on the student-led TBL. Responses to close-ended questions were analyzed using descriptive statistics, and those to open-ended questions were categorized into common themes. RESULTS: A total of 114 learners and 3 student facilitators responded to the questions. Learners found student-led TBL to be just as or more effective than faculty-led TBL in three respects: comprehension (93.0%), active participation (96.5%), and expectation of academic performance improvement (93.9%). According to student facilitators, it improved their knowledge, confidence, communication skills, and leadership abilities. Learners and facilitators indicated that student-led TBL was significantly more effective than faculty-led TBL. Thus, student-led TBL can enhance the ability of all students at different academic levels. DISCUSSION: Student-led TBL appears to be an effective learning strategy in higher education and further shifts toward student-centered learning in the course curriculum.

A rotating cerium anode X-ray system allows visualization of intramural coronary vessels after cardiac stem cell therapy for myocardial infarction.

Conventional angiography is insufficient for evaluating the therapeutic effect of cardiac regeneration therapy. A microangiographic X-ray system using a cerium anode was developed. Cerium has a characteristic X-ray with a peak at 34.6 keV, which allows visualization of tiny amounts of iodine. The performance of the cerium anode X-ray system was evaluated in two excised normal canine hearts and in excised ischemic canine hearts treated with c-kit-positive cardiac stem cells (5 canines) or without cells (5 control canines). In the normal canines, branches penetrating from the left anterior descending artery into the myocardium were visualized, down to third-order branches. In just the treated hearts treated with stem cells, small vessels characterized by irregular vessel walls were observed. The cerium anode X-ray system allowed visualization of microvessels in excised ischemic canine hearts, and may evaluate the effect of cardiac stem cell therapy.

Common and specific risk factors for ischemic stroke in elderly: Differences based on type of ischemic stroke and aging.

BACKGROUND: The risk factors among the types of ischemic stroke (atherothrombotic cerebral infarction: ATI, cardio-embolic infarction: CEI, lacunar infarction: LI) in aged stroke patients have rarely been compared to each other. METHODS: We compared the clinical parameters of 300 elderly patients with ischemic stroke, age 65-98years, to 100 age-matched control patients. RESULTS: Comparison by parametric test and logistic regression analysis between all 300 and 100 control patients showed higher systolic and diastolic blood pressures (p<0.001, p=0.03), lower estimated glomerular filtration rate (eGFR) (p=0.01), larger cardiothoracic ratio (CTR) (p<0.001), smoking (p<0.01) and possibly poor adherence to anti-hypertensive agents in the ischemic stroke patients (p<0.001). Comparisons among three types (n=100 for each) showed the highest atheromatous risk factors for ATI to be hemoglobin A1c (p=0.01) and low-density lipoprotein (p<0.001) and for CEI to be largest cardiac load, indicated by largest left atrial dimension (p<0.001), and CTR (p<0.001). Triglyceride level was found to be a borderline risk factor for LI (p=0.054). Comparison between those aged <74 versus ≥75years (n=150 for each) showed a lower eGFR (p=0.02) and larger right atrial dimension (p<0.001) in patients ≥75. CONCLUSION: The risk factors were quite different among the subtypes and aging.

Evaluation of Intracranial Microvessel Visualization in Mouse and Dog Models by Using a New Rotating Cerium Anode X-ray System.

OBJECTIVE: Lacunar stroke may be caused by infarction of small perforating branches of the middle cerebral artery. We developed a microangiographic X-ray system using a cerium anode to evaluate the perforating branches. METHODS: Iodine has K-edges at 33.2 kilo electron volts. Cerium yields a characteristic X-ray of 34.6 kilo electron volts, therefore, the cerium anode X-ray system could detect tiny amounts of contrast material. First, an X-ray chart was used to evaluate the resolution. Second, the brains of mice were dissected and irradiated. Third, the brains of dogs were excluded and irradiated. Fourth, iodine was perfused into the carotid artery of living dogs during brain imaging. RESULTS: In the first experiment, the cerium anode X-ray system elicited 4.86 clear line pairs. In mice, the perforating branches of the middle cerebral artery could be visualized. The perforating branches were clearly observed in dog brains ex situ even through an acrylic plate, but not in conventional X-ray images. Iodine moving inside the perforating branches was visualized in dog brains in situ using the cerium anode X-ray system. CONCLUSION: The cerium anode X-ray system allowed us to visualize the perforating branches of the middle cerebral artery in living dogs.

Cilostazol protects against microvascular brain injury in a rat model of type 2 diabetes.

Cilostazol, a pluripotent phosphodiesterase III-specific inhibitor with anti-platelet and vasculogenic effects, is useful for preventing recurrent brain vascular events, particularly in stroke patients with diabetes mellitus (DM). However, it is unclear whether cilostazol affects autoregulatory responses in small cerebral arteries. Thus, we investigated the effect of cilostazol on diabetic brain vasculopathy in a model of type II DM using male OLETF rats. OLETF rats were treated with either cilostazol (CG) or vehicle (VG) and subjected to microangiography with monochromatic synchrotron radiation to investigate middle cerebral artery (MCA) vasoreactivity following an injection of acetylcholine (Ach). Ach administration led to MCA diameter contraction in the VG, but MCA dilation in the CG. We also evaluated morphological changes in the small intracranial vessels and found that in the CG, the endothelial cell structure in the small artery was not destroyed. Moreover, protein levels of phosphorylated endothelial nitric oxide synthase (eNOS) and vascular endothelial growth factor (VEGF) were higher in each evaluated brain region in CG rats vs. VG rats. Our results suggest that cilostazol could potentially improve autoregulatory responses in the small cerebral arteries by increasing eNOS phosphorylation and VEGF expression in DM, and thus, may act as a neurovascular protectant.

The vital prognosis of elderly adults living in a group home in their mid-eighties.

The purpose of the present study is to evaluate the vital prognoses of elderly people in their mid-eighties living in a group home (GH) compared to age- and sex-matched outpatient clinic (OPC) in an observational study conducted over 6 years. We investigated the association between mortality and general, cardiac, and nutritional parameters, including eicosapentaenoic acid (EPA) in 54 GH residents (83 ± 8 years old) and 57 OPC attendees (83 ± 5 years old). Kaplan-Meier curves and Cox proportional hazard ratio analyses were used to assess the association between EPA drug administration and mortality in the GH residents and OPC attendees, respectively. The 54 GH residents had higher mortality and poorer nutritional states, as indicated by lower EPA/arachidonic acid values (median 0.20 vs 0.55, p < 0.001), and BMI under the condition without EPA drug administration (1800 mg daily) than did the OPC group. The significant factors that differed between survivors and deceased in the GH residents and OPC attendees were nutritional and cardiac factors. Cox proportional hazard ratio analysis confirmed that a possible determinant of the prognosis was a lower incidence of EPA drug administration and lower hemoglobin in GH. Kaplan-Meier curves and Cox proportional hazard ratio analyses revealed that EPA drug administration significantly reduced the relative mortality by 82 % in the GH residents (p < 0.001) but not in the OPC attendees. The vital prognosis in individuals from GHs was potentially improved by EPA drug administration, which was not the case in the OPC group; however, further prospective studies are needed.

Nutrition-dependent eicosapentaenoic acid deficiency in care house residents.

OBJECTIVE: We have identified that vital prognosis of adults in a group home (GH) in their mid-80s is associated with lower eicosapentaenoic acid (EPA) levels. We investigated the cause of EPA deficiency in residents in a GH and the effect of EPA treatment on cardiac function and nutritional function in 130 older adults. METHODS: We compared blood chemistry data among three age- and sex-matched groups-outpatient clinic (OPC) attendees (n = 54, 87 ± 5 y old), GH residents (n = 40, 85 ± 8 y old), and older adults in a geriatric welfare home for the elderly (GWHE) (n = 36, 87 ± 6 y old)-using non-parametric test. Furthermore, we investigated the sequential changes in blood chemistry and cardiac function at 4 to 12 mo after the initiation of EPA administration (1800 mg/d). RESULTS: Non-parametric test revealed that the EPA/arachidonic acid ratio as well as EPA levels were lower in the GH and GWHE residents than in the OPC attendees (OPC: 0.56 ± 0.3, GH: 0.23 ± 0.12, GWHE: 0.31 ± 0.1). Fish consumption was lower in the GH and GWHE group than in the OPC group. Repeated measured analyses using analysis of variance revealed that EPA administration increased serum EPA levels (54.0 ± 29.0 to 210.5 ± 50.6 µg/mL, P < 0.001); decreased arachidonic acid, docosahexaenoic acid, triacylglycerol, and LDL cholesterol levels at 4.5 ± 3.4 mo after administration; and reduced the severity of supraventricular arrhythmias on ambulatory electrocardiogram at 12.5 ± 4.5 mo (P < 0.05). CONCLUSION: EPA deficiency in GH residents could be related to the nutritional characteristics of older adults in care facilities. EPA treatment induced changes in various lipids and reduced the severity of supraventricular arrhythmias.

Severity of white matter hyperintensities in older adults aged in their mid 80s and its relationship with hemodynamic and cardiac parameters.

AIM: In the present study, we analyzed the severity of white matter hyperintensities detected on magnetic resonance imaging of the brain in older Japanese adults who were aged in their mid 80s, and examined its relationships with the clinical parameters. METHODS: To identify factors related to the severity of white matter hyperintensities in 33 older adult attendees of our outpatient clinic and 17 older adults living in a group home, we carried out logistic regression analyses and/or correlation analyses. RESULTS: Cognitive function and activities of daily living were significantly correlated with the severity of white matter hyperintensities. Multivariate analysis identified activities of daily living, but not cognitive function, as being independently associated with the severity of white matter hyperintensities. Several hemodynamic and cardiac parameters, including diastolic blood pressure, hemoglobin, serum level of N-terminal pro-brain natriuretic peptide, cardiothoracic ratio on the chest X-ray, severity of supraventricular arrhythmias on a Holter electrocardiogram and serum levels of docosahexaenoic acid, were significantly correlated with the severity of white matter hyperintensities. In contrast, the serum cholesterol, glycosylated hemoglobin value and systolic blood pressure were not correlated with the severity of white matter hyperintensities. CONCLUSIONS: The severity of white matter hyperintensities detected on magnetic resonance imaging of the brain in older Japanese patients aged in their mid 80s was significantly correlated with activities of daily living, hemodynamic and cardiac parameters, and the serum level of docosahexaenoic acid.

Altered blood flow in cerebral perforating arteries of rat models of diabetes: A synchrotron radiation microangiographic study toward clinical evaluation of white matter hyperintensities.

AIM: As altered blood flow in the cerebral perforating arteries (PA) might be related to development of cerebral white matter hyperintensities, we examined whether the hemodynamic relationship of the PA and middle cerebral artery (MCA) is altered in rat models of diabetes, compared with normal rats and a rat model of sinoatrial denervation (blood pressure fluctuation model). METHODS: We used microangiography with monochromatic synchrotron radiation to measure the diameters of the PA and MCA at 4.5 µm resolution in five groups of rats: (i) Long-Evans Tokushima Otsuka (LETO); (ii) Otsuka Long-Evans Tokushima Fatty (a model of type 2 diabetes with obesity); (iii) LETO with sinoaortic denervation (LETO + SAD); (iv) F344; and (v) F344 + streptozotocin (a model of type 1 diabetes). RESULTS: Compared with LETO, Otsuka Long-Evans Tokushima Fatty rats showed a significant reduction in the diameter of both PA and MCA, though the PA/MCA diameter ratio was unchanged. In contrast, compared with LETO, LETO + SAD rats showed an increased MCA diameter, and the PA/MCA diameter ratio was decreased. Compared with F344 rats, the MCA diameter was increased in F344 + streptozotocin rats, and the PA/MCA diameter ratio was decreased. Scatter diagrams showed that the diameters of the PA and MCA were essentially independent of each other in the two types of diabetic models. CONCLUSION: PA were consistently visualized at high resolution by means of microangiography using synchrotron radiation. The present results show that rat diabetic models exhibit changes in PA diameter and PA/MCA diameter ratio, which might be related to the development of diabetes-associated cerebral white matter hyperintensities.

Development of Fingertip Synchrotron Radiation Microangiography toward Clinical Prediction of Diabetic Microangiopathy.

OBJECTIVES: The spatial resolution of conventional angiographic systems is not enough to predict diabetic microangiopathy in arterioles (20-200 µm). METHODS: To determine whether fingertip synchrotron (SR) radiation microangiography has enough spatial resolution to quantitate arteriolar diameter changes, and whether an arteriolar paradoxical vasoconstriction is a characteristic observation for diabetic microangiopathy, diameter reduction as arteriolar branching and difference of the diameter changes induced by acetylcholine between control (n = 5) and diabetic rats (n = 5) were analyzed. RESULTS: Fingertip SR microangiography visualized the arterioles with a diameter range of 30-300 µm and demonstrated vascular diameter reduction as branching with a fixed ratio (r = 0.93, P < 0.004 and r = 0.73, P < 0.001). A vasodilatory reaction was induced by acetylcholine in the control (142.4 ± 61.9 to 190.9 ± 73.5, P < 0.05, n = 25), in contrast, paradoxical vasoconstriction in diabetic rats (201.6 ± 83.0 to 16 0.4 ± 67.9, P < 0.05, n = 37). Histological angiopathy was noted only in the diabetic rats. CONCLUSION: In conclusion, the fingertip SR microangiography is useful to predict diabetic micrangiopahty.

Nicorandil prevents sirolimus-induced production of reactive oxygen species, endothelial dysfunction, and thrombus formation.

Sirolimus (SRL) is widely used to prevent restenosis after percutaneous coronary intervention. However, its beneficial effect is hampered by complications of thrombosis. Several studies imply that reactive oxygen species (ROS) play a critical role in endothelial dysfunction and thrombus formation. The present study investigated the protective effect of nicorandil (NIC), an anti-angina agent, on SRL-associated thrombosis. In human coronary artery endothelial cells (HCAECs), SRL stimulated ROS production, which was prevented by co-treatment with NIC. The preventive effect of NIC on ROS was abolished by 5-hydroxydecanoate but not by 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one. NIC also inhibited SRL-induced up-regulation of NADPH oxidase subunit p22(phox) mRNA. Co-treatment with NIC and SRL significantly up-regulated superoxide dismutase 2. NIC treatment significantly improved SRL-induced decrease in viability of HCAECs. The functional relevance of the preventive effects of NIC on SRL-induced ROS production and impairment of endothelial viability was investigated in a mouse model of thrombosis. Pretreatment with NIC inhibited the SRL-induced acceleration of FeCl3-initiated thrombus formation and ROS production in the testicular arteries of mice. In conclusion, NIC prevented SRL-induced thrombus formation, presumably due to the reduction of ROS and to endothelial protection. The therapeutic efficacy of NIC could represent an additional option in the prevention of SRL-related thrombosis.

Low plasma eicosapentaenoic acid concentration as a possible risk factor for intracerebral hemorrhage.

N-3 fatty acids, including eicosapentaenoic acid (EPA), prevent ischemic stroke. The preventive effect has been attributed to an antithrombic effect induced by elevated EPA and reduced arachidonic acid (AA) levels. However, the relationship between intracranial hemorrhage and N-3 fatty acids has not yet been elucidated. In this cross-sectional study, we compared common clinical and lifestyle parameters between 70 patients with intracranial hemorrhages and 66 control subjects. The parameters included blood chemistry data, smoking, alcohol intake, fish consumption, and the incidences of underlying diseases. The comparisons were performed using the Mann-Whitney U test followed by multiple logistic regression analysis. Nonparametric tests revealed that the 70 patients with intracerebral hemorrhages exhibited significantly higher diastolic blood pressures and alcohol intakes and lower body mass indices, high-density lipoprotein (HDL) cholesterol levels, EPA concentrations, EPA/AA ratios, and vegetable consumption compared with the 66 control subjects. A multiple logistic regression analysis revealed that higher diastolic blood pressure and alcohol intake and lower body mass index, HDL cholesterol, EPA/AA ratio, and vegetable consumption were relative risk factors for intracerebral hemorrhage. High HDL cholesterol was a common risk factor in both of the sex-segregated subgroups and the <65-year-old subgroup. However, neither EPA nor the EPA/AA ratio was a risk factor in these subgroups. Eicosapentaenoic acid was relative risk factor only in the ≥65-year-old subgroup. Rather than higher EPA levels, lower EPA concentrations and EPA/AA ratios were found to be risk factors for intracerebral hemorrhage in addition to previously known risk factors such as blood pressure, alcohol consumption, and lifestyle.

Visualization of microvessels by angiography using inverse-Compton scattering X-rays in animal models.

The fundamental performance of microangiography has been evaluated using the S-band linac-based inverse-Compton scattering X-ray (iCSX) method to determine how many photons would be required to apply iCSX to human microangiography. ICSX is characterized by its quasi-monochromatic nature and small focus size which are fundamental requirements for microangiography. However, the current iCSX source does not have sufficient flux for microangiography in clinical settings. It was determined whether S-band compact linac-based iCSX can visualize small vessels of excised animal organs, and the amount of X-ray photons required for real time microangiography in clinical settings was estimated. The iCSX coupled with a high-gain avalanche rushing amorphous photoconductor camera could visualize a resolution chart with only a single iCSX pulse of ∼3 ps duration; the resolution was estimated to be ∼500 µm. The iCSX coupled with an X-ray cooled charge-coupled device image sensor camera visualized seventh-order vascular branches (80 µm in diameter) of a rabbit ear by accumulating the images for 5 and 30 min, corresponding to irradiation of 3000 and 18000 iCSX pulses, respectively. The S-band linac-based iCSX visualized microvessels by accumulating the images. An iCSX source with a photon number of 3.6 × 10(3)-5.4 × 10(4) times greater than that used in this study may enable visualizing microvessels of human fingertips even in clinical settings.

Protective effect of tryptophan against dextran sulfate sodium- induced experimental colitis.

BACKGROUND/AIMS: Little is known about the anti-colitis effect of tryptophan or its metabolites. Here, the protective effect and its mechanism of tryptophan administration on dextran sulfate sodium -induced colitis in mice was studied. MATERIALS AND METHODS: Twenty C57black6 female mice were equally divided into the control group, and treatment group. The control group received a standard CE-2 diet, while the tryptophan group received a CE-2 diet containing 0.5% l-tryptophan. After one week on this diet, all mice were orally administered a solution of 3.5% dextran sulfate sodium for 12 days to induce colitis. Changes in body weight and bloody stool frequency were monitored during dextran sulfate sodium administration. At 12 days post initial dextran sulfate sodium administration, all mice were sacrificed and the histology of their colonic tissue was examined. The nitrotyrosine levels in colonic tissues in both groups, and nitrate and nitrite levels in the urine of the control group, the tryptophan group and the group of mice without dextran sulfate sodium administration was measured. RESULTS: The tryptophan group showed significantly attenuated body weight loss, bloody stool frequency and ameliorated histological changes of colitis. While tryptophan treatment significantly reduced nitrotyrosine level in the colonic tissues, there was no significant reduction in urine nitrate and nitrite levels compared with the (dextran sulfate sodium-induced) control group. CONCLUSION: Tryptophan treatment ameliorated dextran sulfate sodium-induced colitis in this study. One of the anti-colitis mechanisms of tryptophan treatment is attributable to an anti-nitration effect, and may not be via the suppression of nitric oxide generation.

Comparison of eicosapentaenoic acid concentrations in plasma between patients with ischemic stroke and control subjects.

OBJECTIVE: ω-3 fatty acids, including eicosapentaenoic acid (EPA), prevent ischemic stroke. However, the clinical importance of EPA for ischemic stroke and its subtype has not been fully elucidated. METHODS: In a cross-sectional study, we determined whether ω-3 fatty acids were predictive factors for ischemic stroke. We compared common clinical parameters among 65 patients with ischemic stroke and 65 control subjects. The parameters included blood chemistry data; concentrations of EPA, docosahexaenoic acid, and arachidonic acid (AA); EPA/AA ratio; smoking; alcohol intake; fish consumption more than four times per week; and the incidence of underlying diseases. The comparisons were performed using the Mann-Whitney U test, and multiple logistic regression analysis was applied to the significant factors in the non-parametric test. We also applied the same approach to the ischemic stroke subtypes, cardioembolism and large-artery atherosclerosis. RESULTS: In the multiple logistic regression analysis after the Mann-Whitney U test, a lower EPA concentration was one of the significant risk factors for ischemic stroke, as were a lower body mass index, lower high-density lipoprotein cholesterol, and smoking (sensitivity 0.846, specificity 0.831, positive predictive value 0.833). In the analysis of subtypes, a lower EPA/AA ratio and a lower body mass index were the significant risk factors for cardioembolism (sensitivity 0.800, specificity 0.733, positive predictive value 0.750). However, large-artery atherosclerosis was not related to the EPA concentration or the EPA/AA ratio. CONCLUSIONS: In this study, the plasma EPA concentration and the EPA/AA ratio were potential predictive risk factors for ischemic stroke, especially for cardioembolism. Further prospective studies are necessary.

Investigation into bacteremia and spontaneous bacterial peritonitis in patients with liver cirrhosis in Japan.

BACKGROUND/AIMS: Liver cirrhosis patients tend to develop bacteremia, but relatively little has been reported about the situation in Japan. Therefore, we investigated the microorganisms involved and the outcome among liver cirrhosis patients with bacteremia, including spontaneous bacterial peritonitis. METHODS: The subjects were 236 patients who underwent a total of 377 blood cultures and 30 patients diagnosed with spontaneous bacterial peritonitis, among 6,096 inpatients with liver cirrhosis admitted to several hospitals during the period 1991-2009. RESULTS: The rate of positive blood culture was 37.1% (140/377), and the isolated microorganisms were predominantly Gram-negative bacteria. In patients with confirmed bacteremia, the Child-Pugh score and serum blood urea nitrogen and creatinine levels were significantly higher than in non-bacteremia cases. Moreover, short-term mortality (within 1 month) was 48.2% (53/110), being significantly higher than that among non-bacteremia cases (18.8%; 22/117). Among spontaneous bacterial peritonitis cases, mortality within one month was 33.3% (10/30). Again, the Child-Pugh score and serum blood urea nitrogen and creatinine levels were significantly higher among the fatalities than among survivors. CONCLUSIONS: These results indicate that severity of liver dysfunction and severity of renal dysfunction are both important determinants of short-term mortality among liver cirrhosis patients with bacteremia and spontaneous bacterial peritonitis in Japan.

Therapeutic effects of four molecular-weight fractions of Kurozu against dextran sulfate sodium-induced experimental colitis.

BACKGROUND/AIMS: Kurozu has been reported to ameliorate colitis in mice and to have an anti-oxidative effect. However, the active components and mechanism of action remain unknown. Here, as a first step to identify the active components, we chromatographically fractionated Kurozu and investigated the anti-colitis activity of the fractions, focusing on anti-nitration activity. METHODS: Kurozu was divided into 4 molecular-weight fractions (fraction I, >4,000 daltons; II, 2,000~4,000 daltons; III, 800~2,000 daltons; IV, <800 daltons). Forty C57black6 mice were divided into 5 groups as follows: the control group received standard CE-2 diet, and Groups I~IV received CE-2 diet containing Kurozu fractions I~IV, respectively. Dextran sulfate sodium was administered to the mice for 12 days to induce colitis. Body weight and bloody stool frequency were monitored as indices of severity of colitis after administration of dextran sulfate sodium, and at 12 days, all mice were sacrificed for examination of colonic pathology and nitrotyrosine production in the colon tissues. RESULTS: Colitis was markedly ameliorated in Group III, followed by Group II, while Group IV showed little difference from the control. The colonic nitrotyrosine level in Group III was significantly reduced compared with the control. CONCLUSIONS: The major protective components in Kurozu appear to have molecular weights in the range of 800~4,000 daltons, and their action appears to be related, at least in part, to anti-oxidative and anti-nitration effects.

Protective effects of fermented rice vinegar sediment (Kurozu moromimatsu) in a diethylnitrosamine-induced hepatocellular carcinoma animal model.

Kurozu moromimatsu is the sediment of Kurozu, a jar-fermented Japanese black vinegar produced from unpolished rice. Here, we examined the protective effects of Kurozu moromimatsu in a diethylnitrosamine-induced model of hepatocellular carcinoma. Thirty-two F344 rats were divided into two groups; the control group received basal CE-2 diet, and the Kurozu moromimatsu group received CE-2 diet containing Kurozu moromimatsu. At 16 weeks after initial intraperitoneal administration of diethylnitrosamine (150 mg/kg/week), serum was collected from half the rats. These rats were sacrificed and the liver was resected for histological examination of hematoxylin-eosin-stained sections and assay of matrix metalloproteinase-2 and matrix metalloproteinase-9 levels in tumor tissues. Glutathione S-transferase placental form-positive foci were evaluated by immunostaining for glutathione S-transferase placental form. The remaining rats were maintained for evaluation of survival. There were no significant differences of serum transaminases, tumor necrosis factor-alpha, and also no marked hepatic histological differences, between the two groups. However, the size of hepatocellular carcinomas was greatly decreased and the levels of activated matrix metalloproteinase-2 and -9 were significantly reduced in the Kurozu moromimatsu group. Further, survival was significantly prolonged in the Kurozu moromimatsu group compared with the control. These results indicate that Kurozu moromimatsu inhibited the growth of hepatocellular carcinoma.