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Background Slow-motion training, which comprises exercising using extremely slow-movements, yields a training effect like that of high-intensity training, even when the applied load is small. We developed a slow-training exercise program that allows elderly people to safely use their own body weight without a machine. Previously, it was confirmed that functional gait and lower limb muscle strength were improved by low-intensity training using bodyweight training for 3 months. This study evaluated the long-term effects of low-intensity training using body weight with slow-movements on the motor function of frail, elderly patients. Methods Ninety-six elderly men and women aged 65 years or older whose level of nursing care was classified as either support required (1 and 2) or long-term care required (care levels 1 and 2) volunteered to participate. Two facilities were used. Participants at the first facility used low-intensity training using body weight with slow-movements (low-stress training [LST] group, n = 65), and participants at another facility used machine training (MT group, n = 31). Exercise interventions were conducted for 12 months, once or twice per week, depending on the required level of nursing care. Changes in motor function were examined. Results Post-intervention measurements based on the results of the chair-stand test after 12 months showed significant improvements from pre-intervention levels (P

Background Slow-motion training, an exercise marked by extremely slow movements, yields a training effect like that of a highly intense training, even when the applied load is small. This study evaluated the effects of low-intensity bodyweight training with slow movement on motor function in frail, elderly patients. Methods Ninety-seven elderly men and women aged 65 years or older, whose level of nursing care was classified as either support required (1 and 2) or long-term care required (care level 1 and 2), volunteered to participate. Two facilities were used. Participants in the first facility used low-intensity bodyweight training with slow movement (the LST group, n = 65), and participants in another facility used machine training (the control group, n = 31). Exercises were conducted for 3 months, once or twice a week, depending on the required level of nursing care. Changes in motor function were examined. Results Post-exercise measurements showed significant improvements from the pre-exercise levels after 3 months, based on the results of the Timed Up and Go test (p = 0.0263) and chair-stand test (p = 0.0016) in the low-intensity exercise with slow movement and tonic force generation (LST) group. Although the ability to stand on one leg with eyes open tended to improve, no significant change was found (p = 0.0964). Conclusions We confirmed that carrying out LST bodyweight training for 3 months led to improvements in ambulatory function and lower-limb muscle strength. In this way, it is possible that LST training performed by holding a bar or by staying seated on a chair contributes to improved motor function in elderly patients within a short time. Trial registration UMIN000030853. Registered 17 January 2018. (retrospectively registered).

Cassava pulp containing 60% starch and 20% cellulose is a promising renewable source for bioethanol. The starch granule was observed to tightly bind cellulose fiber. To achieve an efficient degradation for cassava pulp, saccharification tests without pre-gelatinization treatment were carried out using combination of commercial α-amylase with cellulosome from Clostridium thermocellum S14 and β-glucosidase (rCglT) from Thermoanaerobacter brockii. The saccharification rate for cassava pulp was shown 59% of dry matter. To obtain maximum saccharification rate, glucoamylase (GA) from C. thermocellum S14 was supplemented to the combination. The result showed gradual increase in the saccharification rate to 74% (dry matter). Supplementation of GA to the combination of commercial α-amylase, cellulosome and rCglT is powerful method for efficient saccharification of cassava pulp without pretreatment.

Application of microbial enzymes for paper deinking is getting tremendous attention due to the rapidly increasing of waste paper every year. This study reports the deinking efficiency of laser-printed paper by the lignocellulolytic enzyme from Penicillium rolfsii c3-2(1) IBRL strain compared to other enzyme sources as well as commercial available enzymes. High enzymatic deinking efficiency of approximately 82 % on laser-printed paper was obtained by pulp treatment with crude enzyme from P. rolfsii c3-2(1) IBRL. However, this crude enzyme was found to reduce the paper strength properties of the pulp based on the results of tensile, tear and burst indices, most probably due to the cellulose degradation. This was further proven by the low viscosity of paper pulp obtained after enzymatic treatment and increasing of sugar production during the treatment. Balancing to this detrimental effect on paper pulp, high deinking efficiency was achieved within a short period of time, in which the enzymatic treatment was conducted for 30 min that enabled contribution to higher brightness index obtained, thus promoting savings of time and energy consumption, therefore environmental sustainability. Extensive research should be conducted to understand the nature and mechanism of enzymatic deinking process by the crude enzyme from P. rolfsii c3-2(1) IBRL in order to improve paper strength properties.

For efficient utilization of both starchy and cellulosic materials, oil palm trunk was separated into parenchyma (PA) and vascular bundle (VB). High solid-state simultaneous saccharification and fermentation (HSS-SSF) using 30% (w/v) PA, containing 46.7% (w/w) starch, supplemented with amylases and Saccharomyces cerevisiae K3, produced 6.1% (w/v) ethanol. Subsequent alkali-pretreatment using sodium hydroxide was carried out with starch-free PA (sfPA) and VB. Enzymatic digestibility of 5% (w/v) pretreated sfPA and VB was 92% and 97%, respectively, using 18 FPU of commercial cellulase supplemented with 10 U of Novozyme-188 per gram of substrate. Likewise, HSS-SSF using 30% (w/v) alkali-pretreated sfPA and VB, with cellulases and yeast, resulted in high ethanol production (8.2% and 8.5% (w/v), respectively). These results show that HSS-SSF using separated PA and VB is a useful fermentation strategy, without loss of starchy and cellulosic materials, for oil palm trunk.

Clostridium thermocellum is known to produce the cellulosomes with efficient plant cell wall degradation ability. To bring out the maximum cellulolytic ability of the cellulosomes, it is necessary to eliminate the end product inhibition by cellobiose. Combinations of β-glucosidases from thermophilic anaerobic bacteria and Aspergillus niger and C. thermocellum S14 cellulosomes were evaluated for optimization of cellulose degradation. β-Glucosidase (CglT) from Thermoanaerobacter brockii, in combination with cellulosomes, exhibited remarkable saccharification ability for microcrystalline cellulose. When rice straw, soaked in 28% aqueous ammonia for 7 days at 60 °C, was hydrolyzed by an enzyme loading combination of 2 mg cellulosome and 10 units CglT per g glucan, 91% of glucan was hydrolyzed to glucose, indicating roughly1/10 the enzyme load of a Trichoderma reesei cellulase (Celluclast 1.5L) and Novozyme-188 combination is enough for the combination of C. thermocellum S14 cellulosomes and CglT to achieve the same level of saccharification of rice straw.

The number of patients with both hypertension and obesity has been increasing in Japan. Many of these patients may also have insulin resistance. Telmisartan, an angiotensin II receptor blocker (ARB), selectively activates peroxisome proliferatoractivated receptor (PPAR)-gamma, and this effect is considered to markedly improve insulin resistance in obese patients with hypertension. We compared the antihypertensive and insulin resistance-improving effects of telmisartan with those of candesartan and valsartan in this patient population. Twenty-eight elderly patients with an average body mass index (BMI) of 27.1 kg/m2 were enrolled in this 6-month study. Patients were randomly selected to either switch from candesartan or valsartan to telmisartan or to continue with their current ARB. A 75 g oral glucose tolerance test (OGTT) was performed before and after switching, and the effect of telmisartan on the insulin response to glucose loading was investigated. There was no significant difference in blood pressure between the two groups after drug administration, but glucose tolerance significantly improved in the telmisartan group. The hyperinsulin response to glucose loading also significantly improved in those taking telmisartan, as well as homeostasis model assessment of insulin resistance (HOMA-IR). These changes were not observed in the control group. In patients with hypertension and obesity showing insulin resistance, treatment with telmisartan significantly improved the hyperinsulin response to glucose loading. Telmisartan may therefore be beneficial in these patients.

An endocellulase-free multienzyme complex was produced by a thermophilic anaerobic bacterium, Thermoanaerobacterium thermosaccharolyticum strain NOI-1, when grown on xylan. The temperature and pH optima for growth were 60 degrees C and 6.0, respectively. The bacterial cells were found to adhere to insoluble xylan and Avicel. A scanning electron microscopy analysis showed the adhesion of xylan to the cells. An endocellulase-free multienzyme complex was isolated from the crude enzyme of strain NOI-1 by affinity purification on cellulose and Sephacryl S-300 gel filtration. The molecular mass of the multienzyme complex was estimated to be about 1,200 kDa. The multienzyme complex showed one protein on native PAGE, one xylanase on a native zymogram, 21 proteins on SDS-PAGE, and 5 xylanases on a SDS zymogram. The multienzyme complex consisted of xylanase, beta-xylosidase, alpha-L-arabinofuranosidase, beta-glucosidase, and cellobiohydrolase. The multienzyme complex was effective in hydrolyzing xylan and corn hulls. This is the first report of an endocellulase-free multienzyme complex produced by a thermophilic anaerobic bacterium, T. thermosaccharolyticum strain NOI-1.

This study examined production of α-glucosidase inhibitors by Bacillus subtilis B2 in Luria-Bertani (LB) fermentation with okara, soy powder, starch or pectin as additional source of carbon and nitrogen. All the fermentation broths of B. subtilis B2 exhibited gradual increase in α-glucosidase inhibitory activity during the fermentation process with or without supplemented source of carbon or nitrogen. Addition of okara into the LB medium greatly enhanced the strength (nearly twice as much of that without okara supplement) of α-glucosidase inhibitory activity of fermentation broth. The α-glucosidase inhibitory activity of B. subtilis B2 fermentation broth was positively correlated (p0.05) with the bacterial populations grown in LB medium containing okara. Glucose and sucrose were not detected in LB medium during the entire fermentation process and were both reduced drastically in media containing okara, soy powder, starch or pectin after 6days of fermentation. The fermented LB medium containing okara by B. subtilis B2 possessed very strong α-glucosidase inhibitory activity and contained little glucose and sucrose, suggesting that fermentation of B. subtilis B2 in LB added with okara might be considered as a strategy for preparing functional foods for diabetic patients.

Excessive visceral fat area (VFA) is a major risk factor in such conditions as cardiovascular disease. In assessing VFA, computed tomography (CT) is adopted as the gold standard; however, this method is cost intensive and involves radiation exposure. In contrast, the bioelectrical impedance (BI) method for estimating body composition is simple and noninvasive and thus its potential application in VFA assessment is being studied. To overcome the difference in obtained impedance due to measurement conditions, we developed a more precise estimation method by selecting the optimum body posture, electrode arrangement, and frequency. The subjects were 73 healthy volunteers, 37 men and 36 women, who underwent CT scans to assess VFA and who were measured for anthropometry parameters, subcutaneous fat layer thickness, abdominal tissue area, and impedance. Impedance was measured by the tetrapolar impedance method using multi-frequency BI. Multiple regression analysis was conducted to estimate VFA. The results revealed a strong correlation between VFA observed by CT and VFA estimated by impedance (r = 0.920). The regression equation accurately classified VFA > or = 100 cm(2) in 13 out of 14 men and 1 of 1 woman. Moreover, it classified VFA > or = 100 cm(2) or < 100 cm(2) in 3 out of 4 men and 1 of 1 woman misclassified by waist circumference (W) which was adopted as a simple index to evaluate VFA. Therefore, using this simple and convenient method for estimating VFA, we obtained an accurate assessment of VFA using the BI method.

Angiotensin II receptor blockers as a class are reported to act as insulin sensitizers. Of these, telmisartan has been shown to have additional unique peroxisome proliferator-activated receptor-gamma-mediated, insulin-sensitizing properties. In this study, investigators explored the effects of telmisartan on glycemic control and lipid metabolism in hypertensive patients with type 2 diabetes who had switched to telmisartan from another angiotensin II receptor blocker. The study subjects were 42 hypertensive outpatients with type 2 diabetes who were being treated with candesartan 8 mg/d and who agreed to switch to treatment with telmisartan 40 mg/d. Relevant laboratory variables were measured 6 mo before treatment switching, at the time of switching, and 6 mo after switching. No significant differences were noted in blood pressure, body mass index, or glycosylated hemoglobin among subjects before and after therapy was switched. No adverse reactions such as edema or hepatic toxicity were noted. No significant changes in fasting plasma glucose, fasting insulin, HOMA-R (insulin resistance as measured by the homeostasis model), preheparin lipoprotein lipase mass, high-density lipoprotein cholesterol, and free fatty acids were noted. Triglyceride levels were significantly decreased, however, and adiponectin levels were significantly increased (8.1+/-3.1 microg/mL at switching; 8.6+/-3.0 microg/mL 6 mo after switching; P.01) after the switch to telmisartan therapy. Study results show that telmisartan did not affect glycemic control, but it improved lipid metabolism and adiponectin production in patients with type 2 diabetes, suggesting that AT(1)-receptor antagonism and selective peroxisome proliferator-activated receptor-gamma activation by telmisartan combine to account for observed effects on lipid metabolism and adiponectin production.

Objective We investigated the relationship between serum uric acid (SUA) and body fat area, serum lipid level, insulin resistance, and metabolic syndrome in Japanese men. Method We studied 508 Japanese man industrial workers who underwent an annual medical examination and agreed to participate in the CT scanning examination. Body fat area was measured at the umbilical level. Metabolic syndrome was defined by the presence of visceral fat Accumulation (≥100 cm2) accompanied by two or more disorders; dislipidemia, hypertension, and hyperglycemia. Results SUA was positively correlated with visceral fat area, subcutaneous fat area, serum total cholesterol level, serum triglyceride level, the Homeostasis Model Assessment index, and was negatively correlated with the high-density lipoprotein cholesterol level. In multiple regression analysis, the most influential factor for SUA was visceral fat area (p=0.0027), followed by the serum triglyceride level (p=0.0245). We clarified a higher SUA in the metabolic syndrome group as compared with the non-metabolic syndrome group: 6.67±1.14 mg, 6.09±1.14 mg, respectively (p

This study characterizes crude enzymes derived from Penicillium rolfsii c3-2(1) IBRL, a mesophilic fungus isolated from the local soil of Malaysia. Prior to enzyme activity evaluation, P. rolfsii c3-2(1) IBRL was inoculated into a broth medium containing oil-palm trunk residues for the preparation of crude enzymes. Oil-palm trunk residues were optimally hydrolysed at pH5.0 and 50°C. P. rolfsii c3-2(1) IBRL-derived crude enzymes displayed higher thermal stability compared with the commercial enzymes, Celluclast 1.5 L and Acellerase 1500. Moreover, the hydrolysing activities of the P. rolfsii c3-2(1) IBRL-derived crude enzymes (xylan, arabinan, and laminarin) were superior compared to that of Celluclast 1.5 L and Acellerase 1500, and exhibit 2- to 3-fold and 3- to 4-fold higher oil-palm trunk residues-hydrolysing specific activity, respectively. This higher hydrolysis efficiency may be attributed to the weak 'lignin-binding' ability of the P. rolfsii c3-2(1) IBRL-derived enzymes compared to the commercial enzymes.

Mutant Clostridium thermocellum YM72 that produces small-size scaffolding protein CipA (ssCipA) was isolated from wild-type YM4. Sequencing of ssCipA revealed that two domains, cohesin 6 and cohesin 7, were not present. Cellulosome prepared from YM72 exhibited a significant reduction of hydrolysis ability on crystalline celluloses such as Sigmacell type-20 and cellulose from Halocynthia. To investigate this influence in vitro, artificial cellulosomes were assembled as recombinant CipA (rCipA) and ssCipA (rssCipA) using native free-cellulosomal subunits. The cellulosome assembled using rssCipA showed a 1.8-fold decrease in the hydrolysis of crystalline cellulose compared with that of rCipA. However, no significant differences in the hydrolysis of carboxymethylcellulose and acid-swollen cellulose were observed. One protein band was missing from the complex that was assembled using rssCipA (confirmed by native-PAGE). The missing protein was identified as CelJ, which is a major cellulosomal subunit. This suggests that insufficient cooperation of CelJ into the cellulosome results in the significant reduction of hydrolysis toward crystalline cellulose. These results indicate that cohesin 6 and 7 may be responsible for the cooperation of CelJ through cohesin and dockerin interactions, and adequate cooperation of CelJ into the cellulosome is important for significant hydrolysis of crystalline cellulose.

We experienced a case in which 131I-6beta-iodomethyl-19-norcholest-5(10)-en-3beta-ol (131I-adosterol) scintigraphy showed high uptake in the right adrenal gland. We diagnosed functional cortical adenoma because of the finding of 131I-adosterol scintigraphy. However, no positive findings for the existence of cortical adenoma were obtained in other examinations and we performed right adrenalectomy. Unexpectedly, pathological finding showed the right adrenal gland was occupied with a large ganglioneuroma. This is an instructive case in which 131I-adosterol scintigraphy showed abnormal high uptake in the adrenal gland, in spite of the fact that the adrenal gland was occupied by a tumor derived from adrenal medulla.

Objectives: Alzheimer’s disease (AD) is diagnosed by either inspection of the brain perfusion SPECT, or three-dimensional stereotactic surface display (3D-SSP). The purpose was to compare diagnostic performances of these methods.Methods: Sixteen nuclear medicine physicians independently interpreted99mTc-ECD SPECT in one session and SPECT with 3D-SSP in another session without clinical information for 50 studies of AD patients and 40 studies of healthy volunteers. Probabilities of AD were reported according to a subjective scale from 0% (normal) to 100% (definite AD). Receiver operating characteristics curves were generated to calculate areas under the ROC curves (Az’s) for the inspection as well as for an automated diagnosis based on a mean Z value in the bilateral posterior cingulate gyri in a 3D-SSP template.Results: Mean Az for visual interpretation of SPECT alone (0.679 ± 0.058) was significantly smaller than that for visual interpretation of both SPECT and 3D-SSP (0.778 ± 0.060). Az for the automated diagnosis (0.883 ± 0.037) was significantly greater than that for both modes of visual interpretation.Conclusions: 3D-SSP enhanced performance of the nuclear medicine physicians inspecting SPECT. Performance of the automated diagnosis exceeded that of the physicians inspecting SPECT with and without 3D-SSP.

We report the first clinical PET study using [1-methyl-11C]8-dicyclopropylmethyl-1-methyl-3-propylxanthine ([11C]MPDX) for imaging adenosine A1 receptors in the human brain. The binding of [11C]MPDX evaluated quantitatively as the distribution volume by a graphical analysis was high in the striatum and thalamus, and low in the cerebellum. The distribution pattern of [11C]MPDX was coincident with that of adenosine A1 receptors in vitro reported previously, and was different from those of blood flow and [18F]FDG. The [11C]MPDX PET has the potential for mapping adenosine A1 receptors in the human brain.

A 41-year-old woman was hospitalized because of gross hematuria. She was found to have a renal arteriovenous malformation and was treated with transcatheter arterial embolization (TAE). Renal scintigraphy with Tc-99m mercaptoacetyltriglycine was performed before and after TAE. Renal scintigraphy was useful for evaluating renal function in this patient.

Bilateral symmetric striopallidodentate calcinosis, also known as Fahr's disease, is characterized by bilateral calcifications of the basal ganglia, thalami, dentate nuclei of the cerebellum, and the white matter of the cerebral hemisphere. Intracranial calcifications are easily visible as high-density areas on computed tomographic images. On magnetic resonance images, the calcifications exhibit different signal intensities. The differences in signal intensity are thought to be related to the stage of the disease, differences in calcium metabolism, and the volume of the calcium deposit. The moderate reduction of cerebral blood flow in bilateral thalami was also identified using brain SPECT.

Objective: We investigated the changes in125I-iomazenil (125I-IMZ) benzodiazepine receptor (BZR) binding with psychological stress in a rat model.Methods: Six male Wistar rats were placed under psychological stress for 1 hour by using a communication box. No physical stress was not received. 1.85 MBq of125I-IMZ was injected into the lateral tail vein and the rat was killed 3 hours later. Twenty-micrometer-thick sections of the brain were collected and % injected dose per body weight (%ID/BW) of eleven regions (frontal, parietal, temporal, occipital cortices, caudate putamen, accumubens nuclei, globus pallidus, amygdala, thalamus, hippocampus and hypothalamus) were calculated by autoradiography. The %ID/BW of rats which were placed under psychological stress was compared with that of 6 control rats.Results: The %ID/BW of rats which were placed under psychological stress diffusely tended to show a reduction in125I-IMZ-BZR binding. A significant decrease in BZR binding was observed in the hippocampus of the rats which were placed under psychological stress.Conclusion:125I-IMZ-BZR binding tended to decrease throughout the brain.

99mTc-methoxy-isobutyl-isonitrile (99mTc-MIBI) scintigraphy with Digirad 2020tc ImagerTM (2020tc), which was a multi-crystal scintillation camera with solid-state detectors was performed for patients with secondary hyperparathyroidism having autografts of parathyroid glands in the right arm. With the 2020tc camera, three abnormal accumulations were found in the right arm. The images obtained with this camera were superior in resolution to those obtained with a conventional NaI crystal gamma camera (ZLC7500, Siemens, Germany). The next day, resection of autografts of parathyroid glands was done. Four hyperplastic parathyroid glands were resected and all were hyperplastic in pathological findings.

We have synthesized β- d -GlcNAc-(1→4)-β- d -GlcNAc-(1→4)-β- d -GlcNAc-(1→4)- d -GlcN (2) through a partial N-acetylation reaction of chitosan tetramer 1 by a chitin deacetylase from Colletotrichum lindemuthianum ATCC 56676. The compound was purified from the mixture of acetylation products of 1 using cation-exchange column chromatography and amine-adsorption column chromatography, and its structure was estimated by 1H NMR and FABMS analyses. The enzymatic reaction allows a regioselectivity that is hard to achieve by chemical N-acetylation.

We report the synthesis of a novel compound for chitinase assays, p-nitrophenyl 2-acetamido-4-O-(2-amino-2-deoxy-β- d -glucopyranosyl)-2-deoxy-β- d -glucopyranoside [GlcNGlcNAc-pNP] by selective N-deacetylation of p-nitrophenyl 2-acetamido-4-O-(2-acetamido-2-deoxy-β- d -glucopyranosyl)-2-deoxy-β- d -glucopyranoside [(GlcNAc)2-pNP] using a purified chitin deacetylase isolated from Colletotrichum lindemuthianum ATCC 56676. FABMS, 1H NMR, and 13C NMR analyses confirmed the structure of this new compound. This disaccharide derivative can be used to distinguish special chitinases that effectively remove partially deacetylated parts of substrates within a mixture of chitinases which degrades (GlcNAc)2-pNP.

Ventilation and pulmonary perfusion scintigraphy were performed in 7 patients in whom postural change from the sitting position to the supine position decreased partial oxygen pressure in arteries (PaO2) by 15 mmHg or more. Six of these 7 patients were obese. Five patients had organic pulmonary disease or space occupying lesions of the liver. On lateral supine-position images taken by using the continuous inhalation method for 81mKr ventilation scintigraphy, ventilation was reduced in the dorsal area corresponding to a gravity-dependent area, but on pulmonary perfusion scintigrams, there were no marked changes compared to normal adults. The mismatch of ventilation to perfusion may have caused hypoxemia. Reduced ventilation was correlated with reduced PaO2. The distribution of 81mKr bolus gas inhalation suggested closure of the airway in the dorsal area at functional residual capacity (FRC), which means the resting expiratory level, in the supine position.

The bone-seeking radiopharmaceutical Sr-89 has been used as a palliative treatment for patients with bone pain caused by bone metastases. The authors report the results of nine patients (three with prostate cancer, four with breast cancer, one with thyroid cancer, and one with lung cancer) who underwent therapy with Sr-89 chloride for painful bone metastases, and evaluate Sr-89 imaging with bremsstrahlung. Two levels of dosage (1.5 and 2.2 MBq/kg) were used. Sr-89 imaging was performed in seven patients 1 week after injection. Abnormal uptake was seen in all and was consistent with the results of Tc-99m HMDP imaging. Six patients were assessed at 3 months and three patients toward the time they were terminal; 78% (seven of nine) derived some benefit. Two patients had a favorable clinical response and showed improvement on Tc-99m HMDP imaging.

This report describes a novel recycling method utilizing a combination of Clostridium thermocellum cellulosomes and Thermoanaerobacter brockii β-glucosidase (CglT). To recover cellulosomes and CglT through re-binding to additional cellulose, a chimeric CBM3-CglT was created by fusing carbohydrate binding module (CBM3) from the scaffolding protein CipA into the N-terminal region of CglT. When a recycling test using cellulosomes and CBM3-CglT was performed on microcrystalline cellulose, the process was capable of 4 rounds of recycling (1%w/vcellulose/round). Although irreversible absorption of cellulosomes and CBM3-CglT into the residues was observed when ammonia-pretreated rice straw and delignified rice straw was used as substrates, a maximum of 2 and 4 recycling rounds (1%w/vglucan/round) were achieved, respectively, consistent with a 70% saccharification rate. This novel recycling method using cellulosomes and CBM3-CglT has great potential as an effective lignocellulose degradation system.

An obligately anaerobic, cellulolytic-xylanolytic bacterium, designated strain A6T, was isolated from soil of a coconut garden in the Bangkuntien district of Bangkok, Thailand. The strain was Gram-stain positive, catalase-negative, endospore-forming, motile and rod-shaped with a cell size of 0.2–0.3×2.0–3.0 µm. Optimal growth of strain A6T occurred at pH55 °C 9.5, 55 °C. Strain A6T fermented various carbohydrates, and the end products from the fermentation of cellobiose were acetate, ethanol, propionate and a small amount of butyrate. The major cellular fatty acids were iso-C14 : 0 3-OH, iso-C15 : 0, iso-C16 : 0 and C16 : 0. The cell-wall peptidoglycan contained meso-diaminopimelic acid. No respiratory quinones were detected. The DNA G+C content was 30.0 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the strain represented a new phyletic sublineage within the family Clostridiaceae , with T represents a novel species of a new genus, for which the name Cellulosibacter alkalithermophilus gen. nov., sp. nov. is proposed. The type strain of the type species is A6T ( = TISTR 1915T = KCTC 5874T).

In order to develop a method for producing fuel ethanol from cassava pulp using cell surface engineering (arming) technology, an arming yeast co-displaying α-amylase (α-AM), glucoamylase, endoglucanase, cellobiohydrase, and β-glucosidase on the surface of the yeast cells was constructed. The novel yeast strain, possessing the activities of all enzymes, was able to produce ethanol directly from soluble starch, barley β-glucan, and acid-treated Avicel. Cassava is a major crop in Southeast Asia and used mainly for starch production. In the starch manufacturing process, large amounts of solid wastes, called cassava pulp, are produced. The major components of cassava pulp are starch (approximately 60%) and cellulose fiber (approximately 30%). We attempted simultaneous saccharification and ethanol fermentation of cassava pulp with this arming yeast. During fermentation, ethanol concentration increased as the starch and cellulose fiber substrates contained in the cassava pulp decreased. The results clearly showed that the arming yeast was able to produce ethanol directly from cassava pulp without addition of any hydrolytic enzymes.

Regioselective synthesis of 2-substituted 3-nitropyridines was achieved by one-pot reaction of either 4- or 6- substituted 1-methyl-3,5-dinitro-2-pyridones with ketones in the presence of ammonia. The selectivity is interpreted in terms of steric factor of substituent on the pyridone

A cellulolytic and xylanolytic enzyme complex-producing alkalothermoanaerobacterium strain, Tepidimicrobium xylanilyticum BT14 is described. The cell was Gram-positive, rod-shaped and endospore-forming. Based on 16S rRNA gene analysis and various lines of biochemical and physiological properties, the strain BT14 was a new member of the genus Tepidimicrobium. The strain BT14 cells had ability to bind to Avicel, xylan and corn hull. The pH and temperature optima for growth were 9.0 and 60 degrees C, respectively. The strain BT14 was able to use a variety of carbon sources. When the bacterium was grown on corn hulls under an anaerobic condition, a cellulolytic and xylanolytic enzyme complex was produced. Crude enzyme containing cellulase and xylanase of the strain BT14 was active in broad ranges of pH and temperature. The optimum conditions for cellulase and xylanase activities were pH 8.0 and 9.0 at 60 degrees C, respectively. The crude enzyme had ability to bind to Avicel and xylan. The analysis of native-PAGE and native-zymograms indicated the cellulose-binding protein showing both cellulase and xylanase activities while SDS-PAGE zymograms showed 4 bands of cellulases and 5 bands of xylanases. Evidence of cohesin-like amino acid sequence seemed to indicate that the protein complex shared direct relationship to the cellulosome of Clostridium thermocellum. The crude enzyme from the strain BT14 showed effective degradation of plant biomass. When grown on corn hulls at pH 9.0 and 60 degrees C under anaerobic conditions, the strain BT14 produced ethanol and acetate as the main fermentation products.

The hyper-cellulolytic strain ofClostridium thermcellum, YM4, produced about 20-fold more CMCase than did strain JW20 (ATCC 31449). Most part of the difference between their enzyme production resided in the enzyme fraction which was free in the culture fluid but could bind to cellulose. The chief distinction of the cellulolytic system of strain YM4 from that of JW20 was the lack of the polycellulosome.

Three-phase bone scintigraphy has been used for the diagnosis of osteomyelitis in some regions of the body. However, its utility in patients with chronic osteomyelitis of the mandible (COM) has been reported only occasionally and the significance has not been fully examined. The aim of this study was to investigate what can be identified from each phase of the three-phase bone scintigraphy in patients with COM. Three-phase bone scintigraphy using 99mTc-labeled phosphonates was performed [96 s (phase 1), 5 min (phase 2) and 3 h (phase 3)] in 15 patients with COM. An increase in accumulation was regarded as a positive result in visual analysis. We investigated the positive ratio of each phase, including in the classification of the disease type, serum white blood cell count, C-reactive protein value and morbidity period. We also calculated the uptake ratio relative to the contralateral side using mean and maximum counts of the region of interest in semiquantitative analysis and investigated the correlation between the uptake ratios of each phase. The positive ratio of phases 2 and 3 was 100% and higher than that of phase 1 (33.3%, p = 0.0001). Almost all patients with a positive result in phase 1 had the osteolytic type (5/6). The extent of increased uptake on phase 3 images was similar to that of phase 2 images. The uptake ratio of phase 2 correlated with that of phase 3 (mean: r = 0.88, p

Paenibacillus curdlanolyticus B-6 showed effective degradation activities for xylan and cellulose and produced an extracellular multienzyme complex (approximately 1,450 kDa) containing several xylanases and cellulases. To characterize the multienzyme complex, we purified the complex from culture supernatants by four kind of chromatography. The purified multienzyme complex was composed of a 280-kDa protein with xylanase activity, a 260-kDa protein that was a truncated form on the C-terminal side of the 280-kDa protein, two xylanases of 40 and 48 kDa, and 60 and 65 kDa proteins having both xylanase and carboxymethyl cellulase activities. The 280-kDa protein resembled the scaffolding proteins of cellulosomes based on its migratory behavior in polyacrylamide gels and as a glycoprotein. Cloning of the 40-kDa major xylanase subunit named Xyn11A revealed that Xyn11A contained two functional domains which belonged to glycosyl hydrolase family-11 and to carbohydrate-binding module family-36, respectively, and a glycine- and asparagine-rich linker. However, an amino acid sequence similar to a dockerin domain, which is crucial to cellulosome assembly, was not found in Xyn11A. These results suggest that the multienzyme complex produced by P. curdlanolyticus B-6 should assemble by a mechanism distinct from the cohesin-dockerin interactions known in cellulosomes.

Childhood obesity is a medical and social problem in Japan. The number of obese children has been increasing steadily since 1980 according to a survey on school health statistics. However, more simple and reliable criteria for assessment of obesity in children are required. The aim of this study was to establish gender- and age-specific body mass index (BMI) cutoff values for evaluation of obesity in school-aged children. A total of 100,587 elementary school, junior and senior high school students, aged 6 to 17 years, comprised the study population, in whom BMI was measured. The BMI cutoff values were calculated by using the receiver operating characteristic (ROC) analysis in the degree of obesity of 20% or greater as definition of obesity. The validity of the determined cutoff values of BMI was then compared with the Rohrer index, the International Obesity Task Force (IOTF) data and the percentile method. The sensitivity and specificity of the BMI cutoff values were over 93% in both boys and girls of all ages, thereby providing criteria that are highly diagnostic in all ages. The Rohrer Index and BMI 95th percentile values tended to underestimate obesity, while the IOTF criteria tended to overestimate it. In conclusion, the BMI cutoff values determined in the present study enable us to detect obese individuals with extremely high precision, and thus may represent an instrument with superior diagnostic capabilities to those of the existing instruments currently being used for assessment of pediatric obesity.

To investigate changes in free benzodiazepine receptor density in response to repeated, long-term administration of diazepam in epilepsy, we assessed125I-iomazenil (125I-IMZ) binding in a mouse model. El mice were divided into two groups of 12 mice each which received either no diazepam (EI(D[-]) group) or 2 mg/kg of diazepam per week (EI(D[+]) group). Nine ddY mice were used as a control. Once each week from the age of 5 to 19 weeks, the El mice received stimulation to produce epileptic seizures 20 minutes after receiving intraperitoneal injections. At 20 weeks of age, a total dose of 0.37 MBq of125I-IMZ was injected in all mice and their brains were rapidly removed 3 hours later. The incidence of epileptic seizures at the age of 19 weeks and the autoradiograms of the brain were compared. The incidence of epileptic seizures in response to weekly stimulation was significantly lower in the E1(D[+]) group than in the E1(D[-]) group (p < 0.001). The percent injected doses of125I-IMZ per gram of tissue in the cortex, hippocampus and amygdala were significantly lower in the E1(D[+]) group than in the E1(D[-]) group (p < 0.05). The results suggest that diazepam binds competitively to125I-IMZ as an agonist to free benzodiazepine receptor sites in the cortex, hippocampus and amygdala and shows anticonvulsant effect in El mice.

Epidemiological and clinical studies on the lifestyle-related obesity have identified smoking, physical activity and alcohol intake as risk factors for obesity. However, no consensus has yet been reached on the effect of smoking on visceral adiposity. This study was designed to assess whether smoking is associated with the accumulation of visceral fat, glucose and lipid metabolism. The subjects were 450 males aged from 24 to 68 years old, who were examined at the health control center in the regular health check conducted by their company. A self-administered questionnaire was used to ascertain smoking status, daily physical activity and alcohol drinking. The number of Brinkman index as an index for smoking status was positively related to being visceral fat area (VFA). In smokers whose Brinkman index was higher, the percent of subjects with abnormal body mass index, VFA, triglyceride, high density lipoprotein-cholesterol, atherosclerotic index, plasma glucose, immunoreactive insulin, or homeostasis model assessment of insulin resistance (HOMA-IR) was higher than that in non-smokers. When evaluated in terms of age-adjusted odds ratios for incidence of a VFA of 100 cm(2) or greater, alcohol drinking was associated with the highest odds ratio. Smoking, physical inactivity and excessive alcohol drinking were associated with visceral adiposity, and smoking affected glucose and lipid metabolism. In conclusions, these findings suggest that smoking is a risk factor for visceral fat accumulation and deterioration of glucose and lipid metabolism.

In recent years, intra-abdominal visceral fat leads to obesity-related complications. A simple indicator that reflects the mass of visceral fat is also needed to enable practical screening of patients. The present study was designed to establish new body mass index (BMI) criteria of central obesity for male Japanese. The subjects were 516 men aged from 19 to 80 years old who were examined at the physical examination center in the regular health check conducted by their company. Correlations between visceral fat area (VFA) or subcutaneous fat area (SFA) and BMI in the subjects were investigated. Receiver Operating Characteristics (ROC) curve was used to find out the optimal cut-off values of BMI to predict central obesity. We compared the percentile ranks corresponding to VFA of 100 cm2, BMI of 25 kg/m2 and new BMI criteria to check to see whether the present BMI criteria classify correctly Japanese men as central obesity. Further evidence for the effectiveness of BMI for VFA is needed. The correlation coefficient between VFA or SFA and BMI was 0.59 or 0.67, respectively. At the cut-off for BMI that maximized sensitivity and specificity for predicting central obesity was 24 kg/m2. Moreover, the percentile value corresponding to VFA of 100 cm2, BMI of 24 kg/m2, and BMI of 25 kg/m2 was the 53, 50 and 61 percentile, respectively. It is necessary to lower a cut-off point for central obesity from BMI of 25 kg/m2 to 24 kg/m2.

3,4-Dihydro-2H-pyrroles are synthesized from γ, δ-unsaturated oximes by photochemical radical cyclization with 1,5-dimethoxynaphthalene (DMN) as the sensitizer. The cyclization of alkyl ketone O-acetyloximes proceeds via photosensitized electron transfer in the presence of acetic acid, while conjugated oximes of aryl and α,β-unsaturated ketones are cyclized via energy transfer.

Recent reports have shown that radio-guided surgery with an intraoperative gamma probe (IGP) is useful for identifying parathyroid adenomas. The aim of this study was to evaluate the usefulness of IGP mapping in patients with secondary hyperparathyroidism. Seven patients with secondary hyperparathyroidism underwent technetium-99m methoxyisobutylisonitrile (Tc-99m MIBI) scintigraphy immediately before undergoing surgical resection of all parathyroid tissues. We compared the sensitivity of Tc-99m MIBI scintigraphy alone with that of the combination of Tc-99m MIBI scintigraphy and IGP mapping. The sensitivity of the combination of Tc-99m MIBI scintigraphy and IGP mapping was 100% (28 of 28 lesions) and was significantly higher than that of Tc-99m MIBI scintigraphy alone (71.4%, 20 of 28 lesions). The combination of the Tc-99m MIBI scintigraphy and IGP mapping has increased sensitivity for identifying parathyroid lesions during parathyroidectomy in patients with secondary hyperparathyroidism.

Insulin secretion dynamics and response to nateglinide were studied in patients with type 2 diabetes and reduced early-phase insulin secretion. On day 1, 24 patients underwent a 75-g oral glucose tolerance test without taking nateglinide. On day 2, they were given oral nateglinide 90 mg immediately before the oral glucose tolerance test. After glucose loading, insulin levels increased significantly at 30, 60, 90, and 120 minutes after the patients took nateglinide, along with insulinogenic indices, the total area under the insulin curve, the area under the 0- to 90-minute insulin curve, and the area under the 90- to 180-minute insulin curve. Both the plasma glucose level at 60, 90, 120, and 180 minutes and the total area under the glucose curve were significantly reduced following nateglinide administration. Compared with the low responders (n=13), the high responders (n=11) had a significantly shorter duration of disease, significantly higher insulinogenic indices in the absence of nateglinide administration, and a higher homeostasis model assessment-beta cell performance. Nateglinide demonstrated a rapid-onset and rapid-offset insulin secretion-stimulating effect in this study population. A single dose of nateglinide may be indicated for patients with a relatively high homeostasis model assessment-beta cell performance, a short duration of disease, and relatively high insulinogenic indices prior to nateglinide administration.

Thermoanaerobacter thermohydrosulfuricus strain YM3 andClostridium thermocellum strain YM4, obtained originally as a stable coculture, required yeast extract to grow separately. Cell-free broths ofT. thermohydrosulfuricus strain YM3 andC. thermocellum strain YM4 monocultures replaced yeast extract in supporting the growth of strains YM4 and YM3, respectively.T. thermohydrosulfuricus strain YM3 produced vitamin B6, B12 analog(s),p-aminobenzoic acid and folic acid, which were required byC. thermocellum strain, YM4. Likewise, strain YM4 produced niacin-active compound(s), thiamine, and methionine required by strain YM3.

We compared the ability to diagnose skeletal metastasis between serum prostate specific antigen (PSA), C-terminal propeptide of blood type I procollagen (PICP), and urine type I collagen-crosslinked N telopeptide (NTx) in prostate cancer patients. In sixty-nine patients with prostate cancer, bone scintigraphy was performed, and serum PSA and PICP and urine NTx were measured. The median level of serum PSA in the osseous metastasis-negative group (n = 33) was 0.80 ng/ml being significantly lower as compared to the osseous metastasis-positive group (n = 36, 7.70 ng/ml) (p0.0001). The serum PICP and urine NTx/Cr levels appeared lower in the osseous metastasis-negative group than the osseous metastasis-positive group, but there was no significant difference. Logistic regression analysis showed that ability to diagnose skeletal metastasis of serum PSA was 68.1% and superior to those of serum PICP (56.5%) and urine NTx/Cr (53.6%). Serum PSA improved the ability to diagnose skeletal metastasis when combined with serum PICP or urine NTx/Cr. When patients were grouped according to the extent of disease grade (EOD grade) nomenclature, Spearman's correlation coefficient by rank showed that serum PSA was most significantly correlated with EOD grade (p0.0001). In 14 patients whose skeletal metastases progressed or regressed, the change of serum PSA more clearly separated the osseous metastasis-regression group and osseous metastasis-progression group than did serum PICP and urine NTx/Cr. Serum PSA was more reliable than bone resorption and formation markers produced by crosslinking of type I collagen.

The aim of this study is to re-evaluate and clarify the diagnostic role of ventilation/perfusion lung scintigraphy in Japan, now that single-detector-row helical CT and multidetector-row CT are available in clinical practice. The Respiratory Nuclear Medicine Working Group of the Japanese Society of Nuclear Medicine distributed a questionnaire to institutions in Japan equipped with scintillation cameras as of September 2001. Of 1,222 institutions, 239 returned effective answers (19.6%). The most frequent combination for initial diagnosis of acute pulmonary thromboembolism was chest radiography, perfusion lung scintigraphy, and contrast-enhanced CT (111 institutions, 46.4%). The questionnaire revealed that the validity and usage of perfusion lung scintigraphy and those of contrast-enhanced CT were equivalent in the present clinical situation. On the other hand, the diagnostic value of ventilation lung scintigraphy in suspected pulmonary thromboembolism has not been established in Japan. Even though contrast-enhanced CT is widely used in Japan, perfusion lung scintigraphy is still required to determine disease severity and monitor its progress.

A 43-year-old man fell down after drinking alcohol and had an abnormal finding on brain computed tomography in the emergency room. The findings of a neurologic examination were not significant. Cerebral perfusion scintigraphy was performed using Tc-99m L,L-ethylcysteinate dimer (Tc-99m ECD). Dynamic radionuclide angiography showed a site of increased activity in the left cerebral hemisphere. SPECT showed decreased perfusion in the left corpus striatum. MRI showed a left caudate head arteriovenous malformations (AVM). This is a rare case in which dynamic and SPECT Tc-99m ECD images show the presence of an AVM that correlated with MRI.

We found that a chitin deacetylase from Colletotrichum lindemuthianum could acetylate free amino sugar residues into N-acetylated forms in the presence of 3.0 M sodium acetate. The result was analyzed using a β- N -acetylhexosaminidase-coupled assay system with p -nitrophenyl 2-amino-2-deoxy-β- d -glucopyranosyl-(1→4)-2-acetamido-2-deoxy-β- d -glucopyranoside as the substrate, and the liberation of p -nitrophenol was observed as a consequence of enzymatic N-acetylation of the glucosamine residue at the nonreducing end of the substrate. The chitin deacetylase also acetylated chitobiose and chitotetraose as substrates, which was evidenced by the decrease in the amount of free amino sugar residues in the chitooligosaccharides. The reaction product of chitobiose after the acetylation reaction was exclusively 2-acetamido-2-deoxy-β- d -glucopyranosyl-(1→4)-2-amino-2-deoxy- d -glucose [GlcNAcGlcN], the structure of which was determined by FABMS and NMR analyses. This study offers a novel method for enzymatic N-acetylation of amino sugars, and especially with chitobiose as substrate, a selectively N-acetylated product, GlcNAcGlcN, can be synthesized.