Impact of different timing of consuming sweet snack on postprandial glucose excursions in healthy women.

AIMS: Our aim was to evaluate the acute effect of eating sweet snacks at different times of day on glycaemic parameters in young women without diabetes. METHODS: In this randomized controlled three-treatment crossover study, 17 women [(means ± SD) age: 21.2 ± 0.8 years, BMI: 20.7 ± 2.5 kg/m2, HbA1c: 36 ± 2 mmol/mol (5.1 ± 0.2%)] wore flash (continuous) glucose monitoring systems for 7 days. Each participant consumed identical test meals on days 4, 5 and 6, but consumed sweet snacks (baked cake: 498 kcal; 53.6 g of carbohydrate, 8.0 g of protein, 28.0 g of fat) at 12:30 (post-lunch), 15:30 (mid-afternoon) and 19:30 (post-dinner), respectively, on each of those days. Daily glycaemic parameters on those 3 days of snacking at different times of day were compared within-participant. RESULTS: The mean amplitude of glycaemic excursions (3.54 ± 0.32 vs. 2.73 ± 0.20 mmol/L; P < 0.05), standard deviation of glucose (1.20 ± 0.11 vs. 0.92 ± 0.07 mmol/L; P < 0.05), incremental area under the curve (IAUC) for glucose at 12:00-07:00 (986 ± 89 vs. 716 ± 88 mmol/L × min; P < 0.05) and IAUC at 07:00-10:00 the next day (141 ± 17 vs. 104 ± 12 mmol/L × min; P < 0.05) when the snack was eaten post-dinner were all significantly higher than with mid-afternoon snacking. CONCLUSION: Eating sweet snacks post-dinner should be avoided because it worsens glucose excursions as well as postprandial glucose levels after both dinner and the following day's breakfast in young healthy (non-diabetic) women.

Consuming snacks mid-afternoon compared with just after lunch improves mean amplitude of glycaemic excursions in patients with type 2 diabetes: A randomized crossover clinical trial.

AIMS: Our aim was to explore the acute effects of consuming snacks at different times on glucose excursions in patients with type 2 diabetes (T2D). METHODS: Seventeen patients with T2D [means±SD: age 67.4±9.4-years; BMI 23.5±3.1kg/m2; HbA1c 55±6mmol/mol (7.2±1.0%)] were randomly assigned in this crossover study. Each participant wore a continuous glucose monitoring device for 4 days and consumed identical test meals on the second and third days, comprising breakfast at 0700h, lunch at 1200h and dinner at 1900h. Half the participants consumed 75kcal biscuits at 1230h (just after lunch) on the second day and at 1530h (mid-afternoon) on the third day, while the other half consumed snacks at the same times, but vice versa. Each patient's glucose parameters were compared against baseline for the 2days of snacking at different times of day. RESULTS: Consuming snacks in the mid-afternoon led to significantly lower mean amplitudes of glycaemic excursions (mean±SEM: 5.19±0.48 vs. 6.90±0.69mmol/L, P<0.01; standard deviation: 1.75±0.17 vs. 2.16±0.21mmol/L, P<0.01) and incremental areas under the curve for glucose after dinner (479±76 vs. 663±104mmol/L per min, P<0.01) compared with snacking just after lunch, whereas mean glucose levels did not differ over the 2days. CONCLUSION: These results suggest that consuming snacks well separated from lunch may be an effective way to suppress postprandial glucose levels and glycaemic excursions.

Identification of genes induced by taxol application using a combination of differential display RT-PCR and DNA microarray analysis.

The differential display reverse transcriptional polymerase chain reaction (DD-RT-PCR) was used to hunt for cDNA fragments specifically expressed by taxol treatment of HeLa cells. Forty-eight cDNA clones were differentially displayed through the experiments. The cDNA fragments obtained were separately spotted onto glass slides to prepare a tailor-made DNA chip. The gene expression pattern of differentially displayed cDNA fragments were checked by DNA microarray analysis.

SELEX for tubulin affords specific T-rich DNA aptamers. Systematic evolution of ligands by exponeential enrichment.

We succeeded in acquiring two DNA aptamers that selectively recognize tubulin by the SELEX method. A pool of single-stranded oligo-DNAs including a random region of 59 nucleotides was screened by SELEX for tubulin purified from calf-brain as a target. After 20 repetitions of selection round, the library converged on specific T-rich sequences. The binding activity of T-rich clones was analyzed by the SPR sensor to determine their dissociation constants to be in the order of 10 microM.

[Effect of bolus propofol administration on muscle evoked potential (MsEP) during spine surgery].

Intraoperative monitoring of descending pathways by means of muscle evoked potential (MsEP) is a reliable method to monitor spinal cord motor function, but MsEP is readily affected by anesthetics. We monitored MsEP evoked by repetitive transcranial electrical stimulation of the motor cortex in 30 patients receiving spine surgery. Total intravenous anesthesia was maintained with propofol and fentanyl without any muscle relaxant. Onset latencies and peak to peak amplitudes of MsEP were evaluated before and after the bolus propofol administration. The concentrations of propofol in blood and the effect-site during MsEP monitoring were predicted by computer simulation software. The amplitude of MsEP decreased slightly by bolus propofol administration, but the latencies showed no significant change with propofol under the same condition. We consider that total intravenous anesthesia with propofol and fentanyl without muscle relaxants is compatible with the recording of MsEP evoked by high frequency repetitive electrical transcranial stimulations. When MsEP is monitored during spine surgery, anesthetic condition should be controlled carefully in order to maintain a stable blood concentration of propofol and thus to assure the reliability of MsEP measurements.

The occurrence of geometric polyprenol isomers in the rubber-producing plant, Eucommia ulmoides Oliver.

The chain length and geometric isomerism of polyprenols from Eucommia ulmoides Oliver were analyzed using supercritical fluid chromatography. After intensive effort to establish separation conditions for geometric isomers, a phenyl-bonded silica gel-packed column was found that cleanly separated poly-trans and -cis prenols. The presence of long-chain poly-trans prenols (>9 mers) was confirmed for the first time in plants. Trans isomers were found in the leaf, seed coat, and root, but not in the bark and seed. Poly-trans prenols in this plant may act as intermediates for trans-polyisoprene biosynthesis.

High-resolution analysis of polyprenols by supercritical fluid chromatography.

A high-resolution analysis of polyprenol mixtures was achieved by supercritical fluid chromatography (SFC). The separation of polyprenols was examined on an octadecylsilane-packed column with liquid carbon dioxide as the mobile phase and ethanol as modifier. Using this chromatography system, the resolution of separation (Rs) between octadecaprenol (prenol 18) and nonadecaprenol (prenol 19) was two times higher than that using conventional reversed-phase high-performance liquid chromatography. Our SFC technique allows the advantage of baseline separation of polyprenol samples containing hydrophobic components such as terpenes or fatty acids that are unfavorable for good separation. This method is very useful for the analysis of structurally close polyprenol analogues of rubber plant metabolites.

DNA aptamers that bind to chitin.

We have succeeded in the acquisition of DNA aptamers that recognize chitin using in vitro selection. The obtained DNA aptamers have the stem-loop or bulge loop structures with guanine rich loop clusters and the clockwise B-form stems.

HLA-associated cellular response to GAD in type 2 diabetes with antibodies to GAD.

Proliferative response of peripheral blood mononuclear cells (PBMC) to glutamic acid decarboxylase (GAD), which has been reported in patients with type 1 diabetes, was measured in type 2 diabetes, especially in patients with antibodies to GAD initially diagnosed as having type 2 diabetes (anti-GAD+ type 2 diabetes). We studied 12 patients with type 1 diabetes, 22 with anti-GAD+ type 2 diabetes, 31 with type 2 diabetes who were negative for anti-GAD (anti-GAD+ type 2 diabetes), and 30 healthy control subjects for cellular responses in vitro to GAD. The mean stimulation index (SI) in response to GAD was significantly higher in type 1 diabetes than in anti-GAD+ type 2 diabetes or healthy controls (1.47+/-0.35 vs. 1.11+/-0.35, P<0.05, and 1.06+/-0.07, P<0.05, respectively). The mean

An artificial plastic receptor that discriminates axial asymmetry.

An artificial plastic receptor that can discriminate axial asymmetry of optically active binaphthyldiamine derivatives was prepared by the 'molecular imprinting' technique. A light-radical polymerization in the presence of an axially asymmetric compound, (R)-2,2'-bis-methylcarbonylamino-1,1'-binaphthyl (binaphthyldiamine bis-acetamide (BINADA-ac)) as a template molecule with methacrylic acid (MA) as a functional monomer, and ethyleneglycol dimethacrylate (EGDMA) as a crosslinker, at 4 degrees C. The obtained polymer exhibited a superior enantioselectivity to the (R)-enantiomer by HPLC analyses. This plastic receptor should recognize the template molecule with its shape and character of its functional groups. It should be useful in the development of chiral stationary phases for the optical resolutions of axially asymmetric compounds.

A simple assay for formate dehydrogenase activity by gas chromatography-mass spectrometry.

A new method using GC-MS was devised for the convenient measurement of formate dehydrogenase (FDH) activity in crude tissue samples. FDH activity was detected by measuring headspace 13CO2, which was enzymically converted from [13C]formic acid. This method proved to be sensitive and simple for the estimation of FDH activity without complicated pretreatment.

Effects of qigong walking on diabetic patients: a pilot study.

OBJECTIVES: The present study was designed to evaluate the advantages of qigong walking, a mild and slow exercise that uses all the muscles of the body, in comparison with conventional walking in patients with diabetes. INTERVENTIONS: Ten inpatients with diabetes mellitus and associated complications were studied on 3 different days. Either qigong walking (30-40-minute duration) or conventional walking was performed by the patients 30 minutes after lunch on 1 of the 3 study days. Plasma glucose levels and pulse rates were measured 30 minutes after lunch and again 20 minutes after exercising; that is, 90 minutes after lunch. These data were compared to those obtained on a day with no exercise after lunch. RESULTS: Plasma glucose levels decreased during both exercises (from 228 mg/dL before to 205 mg/dL after conventional walking) and (from 223 mg/dL before to 216 mg/dL after qigong walking). In both situations the results after exercise decreased more than those in the group with no exercise (229 mg/dL; p < 0.025). The pulse rates increased after conventional walking (from 77 to 95 beats per minute; p < 0.025) and were higher than those in the group with no exercise (70 beats per minute; p < 0.01) and those after qigong walking (79 beats per minute; p < 0.05). CONCLUSIONS: Qigong walking reduced plasma glucose after lunch without inducing a large increase in the pulse rate in patients with diabetes.

Ratio of motor nerve conduction velocity to F-wave conduction velocity in diabetic neuropathy.

OBJECTIVE: To investigate the usefulness of a new parameter, the ratio of motor nerve conduction velocity to F-wave conduction velocity (M/F ratio), for the differential diagnosis of diabetic neuropathy. RESEARCH DESIGN AND METHODS: Nerve conduction studies were conducted in 95 patients with diabetic neuropathy, 44 nondiabetic patients with peripheral neuropathy, and 24 normal control subjects. Nondiabetic patients with neuropathy were grouped by clinical diagnosis as follows: segmental demyelination (n = 15), axonal neuropathy (n = 11), alcoholic polyneuropathy (n = 4), and other polyneuropathy (n = 14). Motor nerve conduction velocity (MCV) of post-tibial nerves, sensory nerve conduction velocity (SCV) of sural nerves, and F-wave conduction velocity (FWCV) of post-tibial nerves were measured by standardized techniques. The M/F ratio was calculated from these measurements. RESULTS: The MCV and SCV of diabetic patients were significantly slower and the M/F ratio was significantly lower than those of normal subjects: MCV, 43.7 +/- 5.4 vs. 47.1 +/- 2.9 m/s, P < 0.001; SCV, 44.7 +/- 11.1 vs. 48.3 +/- 5.7 m/s, P < 0.05; M/F ratio, 0.84 +/- 0.09 vs. 0.90 +/- 0.06, P < 0.001. The FWCV of nondiabetic patients with neuropathy was significantly slower (40.0 +/- 6.3 vs. 48.3 +/- 4.0 m/s, P < 0.001) and the M/F ratio was significantly higher (1.04 +/- 0.12, P < 0.001) than that of normal subjects, respectively. Although MCV, SCV, and FWCV were correlated with age in normal control subjects, the M/F ratio was independent of age in the diabetic as well as the nondiabetic patients with neuropathy. CONCLUSIONS: Results suggest that the M/F ratio, which is influenced by the neuronal damages in the distal segment of peripheral nerves, is useful in the differential diagnosis of diabetic neuropathy.

[Mild hypothermia anesthesia for carotid microendoarterectomy in a patient with ischemic heart disease].

A 69 year-old male with ischemic heart disease indicated for coronary artery bypass grafting was scheduled for carotid microendoarterectomy. We induced mild hypothermia technique with vasodilation and surface cooling by convecting warming device. We examined hemodynamics by pulmonary artery catheter. Anesthesia was induced with thiamylal, fentanyl, midazolam and isoflurane in nitrous oxide and oxygen. Following administration of vecuronium, trachea was intubated. Pulmonary artery catheter was inserted from the femoral vein. Dopamine, dobutamine 3-5 micrograms.kg-1.min-1 and PGE1 5-10 ng.kg-1.min-1 were continuously administered to keep peripheral blood circulation and cardiac output (CO). Systemic vascular resistance decreased from 1800 to 591 dyne.s.cm-5 and CO increased from 2.8 to 6.9 l.min-1. The occlusion of blood flow of the right carotid artery for 40 min at 34.5 degrees C of rectal temperature did not cause any neurological deficits. No other complications such as arrhythmia, myocardial ischemia and bleeding tendency were observed. Keeping peripheral blood circulation and uniform cooling and warming are important in inducing mild hypothermia safely in a patient with ischemic heart disease.

HLA-DRB1 alleles contribute to determining the prognosis of Japanese diabetes mellitus positive for antibodies to glutamate decarboxylase.

Diabetes mellitus positive for antibodies to glutamate decarboxylase is heterogeneous as far as the degree of impairment of endogenous insulin release, though antibodies to glutamate decarboxylase are the most useful marker for future insulin deficiency. To investigate what determines the prognosis of diabetes mellitus positive for antibodies to glutamate decarboxylase, we measured HLA-DRB1 alleles in three groups: 77 cases of insulin-dependent diabetes mellitus (IDDM), 44 of non-insulin-dependent diabetes mellitus (NIDDM) with secondary failure of oral hypoglycemic therapy, and 22 of NIDDM well controlled by diet and/or sulfonylurea agents. The proportion of susceptible and resistant alleles to IDDM determined the degree of insulin deficiency, and comparison of IDDM to NIDDM well controlled by diet and/or sulfonylurea agents revealed significant differences in DRB1*0405 (P < 0.05; RR = 2.82 and RR = 0.89, respectively) and DRB1*1502 (P < 0.001; RR = 0.02 and RR = 2.19, respectively). This study revealed that HLA-DRB1 alleles contribute to determining the prognosis of Japanese diabetes mellitus positive for antibodies to glutamate decarboxylase.

Diagnostic significance of antibodies to glutamic acid decarboxylase in Japanese diabetic patients with secondary oral hypoglycemic agents failure.

Some non-insulin-dependent diabetes mellitus (NIDDM) patients are positive for antibodies to glutamic acid decarboxylase (anti-GAD), and they tend to develop insulin deficiency. The aim of this study was to evaluate the prevalence of anti-GAD in NIDDM with secondary failure of sulfonylurea agents (NIDDM-SF) and to investigate the diagnostic significance of seropositivity for anti-GAD in NIDDM-SF patients by evaluating human leukocyte antigen (HLA)-DRB1 alleles concurrently. The prevalence of anti-GAD in NIDDM-SF, NIDDM, and new-onset (within 1 year after onset) insulin-dependent diabetes mellitus (IDDM) was 9.3% (39/420), 3.1% (12/392), and 65.0% (13/20), respectively. Pancreatic beta cell function deteriorated in NIDDM-SF patients positive for anti-GAD. HLA-DRB1 allele typing revealed that NIDDM-SF patients positive for anti-GAD were significantly associated with DRB1*0901 (RR = 2.81, P < 0.01), which is one of the susceptible alleles to IDDM. Shorter interval before development of secondary failure and insulin deficiency were significantly associated with the presence of DRB1*0901 (P < 0.05) in NIDDM-SF patients positive for anti-GAD. In conclusion, nearly 10% of NIDDM-SF patients are positive for anti-GAD, suggesting that an autoimmune mechanism might play an important role in the pathogenesis of NIDDM-SF patients. In addition, a combination of serological marker (anti-GAD) and genetic marker (HLA-DRB1) is useful for predicting clinical course of NIDDM patients with secondary failure of sulfonylurea agents.

High prevalence of mitochondrial diabetes mellitus in Japanese patients with major risk factors.

To identify diabetes mellitus caused by the mitochondrial gene substitution at genomic nucleotide pair 3243 (M3243A-->G) we selected 87 diabetic patients with high risk factors such as maternal inheritance and hearing loss. Total DNA was extracted from peripheral leukocytes, and mitochondrial DNA fragments containing M3243A-->G were amplified by polymerase chain reaction (PCR). The amplified fragments were digested with a restriction endonuclease Apa1 and analyzed by agarose gel electrophoresis. The incidence of the M3243A-->G mutation was 4.6% (four of 87) in diabetic patients with maternal inheritance and/or hearing loss. In a subgroup with both maternal inheritance and hearing loss, the incidence of the mutation was as high as 21.4% (three of 14). Cardiac disorders were also present in all four diabetic patients with the mutation. This study suggests that maternal inheritance and hearing loss are useful clinical findings to identify diabetic patients with the mutation, and that cardiac involvement is a high risk factor for the M3243A-->G mutation.

[The efficiency of a new porous type leukocyte removal filter for red cell blood components, Terumo Imuguard III-RC, in the rapid transfusion conditions].

We evaluated the efficiency of a new porous type leukocyte removal filter for red cell blood components, Terumo Imuguard III-RC, in the rapid transfusion conditions. One leukocyte removal filter was used for 2 units of RC-M.A.P (red cell mannitol-adenine-phosphate). Filtration methods employed were gravity infusion, high pressure infusion (300 mmHg), pumping infusion and 20 ml.min-1 infusion under high pressure (300 mmHg). Blood samples were taken before and after the filtration to measure white blood cell (WBC), red blood cell (RBC) and platelet content. Blood samples before filtration and after filtration with WBC excluded, were examined by automated hematology analyzer (Coulter counter STKS-Retic). WBC after filtration was counted by the hemacytometer method using Nageotte Chamber. The removal rate of WBC was found to be more than 99.99% and residual WBC content was less than 4 x 10(4) with every method. The recovery rate of RBC was not significantly decreased in all filtration methods. The removal rate of platelet was equal in all filtration methods. In conclusion, Imuguard III-RC could be useful for effective homologous blood transfusion.

Antimicrobial constituents of Angelica dahurica roots.

One novel coumarin from Angelica dahurica roots was elucidated to be 5,8-di(2,3-dihydroxy-3-methylbutoxy)-psoralen. It occurs together with six other known coumarins and ferulic acid. The antimicrobial activity of the coumarins and ferulic acid were compared.