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ABSTRACT Objective Methods Results Conclusion Whether or not an athlete plays with sports equipment in their hands may influence handgrip strength (HGS) changes during development, but longitudinal studies have not confirmed this. This study compared one‐year HGS changes between two sports types (soccer vs. kendo) in children and adolescent athletes.One hundred sixty‐eight young athletes (86 kendo boys and 82 soccer boys) had two HGS measurements separated by 1 year. A 2 (sports) by 2 (timepoints 1 and 2) repeated measures ANOVA was used to determine whether HGS changed differently between sports.There was no evidence for a sport × time interaction in HGS (p = 0.14); however, the mean difference and 95% CI were in the direction of favoring a greater change in kendo athletes [difference of 0.6 (95% CI: −0.2, 1.5) kg]. There was a main effect of time and sport. Kendo athletes had a 4.6 (95% CI: 1.8, 7.5) kg greater HGS than soccer athletes. There was no evidence that the change in HGS between sports depended on the initial age of the athlete (p = 0.205).Using sports equipment during play may positively affect HGS. [ABSTRACT FROM AUTHOR]

Objectives: Males, on average, are bigger and stronger than females. Hormonal differences during puberty are one reason given for this performance advantage. However, not all evidence supports that thesis. Our aim was to further this discussion by measuring early life changes between sexes (when hormones would be similar) in components of muscle function. Methods: Fifty‐one children (29 boys, 22 girls) completed this study. Forearm muscle size and strength were assessed three times with each time point being separated by approximately a year (2021–2023). Results: There was no sex*time interaction for handgrip strength (p = 0.637). There was, however, a time (p < 0.001) and sex (p < 0.001) effect. Strength increased each year and boys were stronger than girls (difference of 1.5 [95% 0.7, 2.3] kg). There was no sex*time interaction for ulnar muscle thickness (p = 0.714) but there was a time (p < 0.001) effect. Muscle size increased each year but there was no evidence of a sex effect (p = 0.12; difference of 0.81 [95% −0.21, 1.8] mm). A strong positive within‐participant correlation between muscle size and strength (r = 0.803 95% CI: [0.72, 0.86], p < 0.0001) was found across time. Conclusion: Muscle size and strength increased together but this increase did not differ based on sex and boys were stronger than girls. Future work is needed to determine the reason for this difference in maximal strength. Any effect was seemingly present at the initial measurement (at the age of 4 years), since muscle size and strength did not change differently between boys and girls over time. [ABSTRACT FROM AUTHOR]

Objective: This study aimed to compare the current handgrip strength (HGS) of Kendo athletes with their HGS when they were in university (up to 50 years). Methods: Eighty male graduates who were Kendo club members during their university days performed anthropometric and HGS measurements, and these HGS were compared with those measured during their university days (mean age of 19.5 years old). Results: There was no evidence of a statistical difference in HGS between the current measurement and the measurement taken during university [−0.64 (−1.9, 0.67) kg, p =.336]. There was, however, evidence that the difference in HGS depended upon the current age of the individual (t = −6.43, p <.001). When probing the interaction, there were statistical differences between the ages of 24.6 and 38.2 years and between the ages of 47.4 and 69.9 years. Strength increased across time in the younger participants and decreased for those who were older. Between the ages of 38.9 and 46.1 years, there was no evidence of a statistical difference indicating a maintenance of strength. Conclusion: The HGS of Kendo club graduates, which they acquired during their formative years, continued to increase even after they graduated from university and entered their 30s. However, their HGS decreased from age 50, even though they practiced Kendo. [ABSTRACT FROM AUTHOR]

Objective: Handgrip strength may differ depending on the type of sport played during the developmental period. Youth sports in which athletes hold equipment in their hands may be the most effective for improving handgrip strength. This study aimed to examine the age at which differences in handgrip strength appear by comparing sports that involve gripping (kendo) with those that do not involve gripping (soccer) in young athletes. Methods: Two hundred and twenty‐two male athletes (115 kendo and 107 soccer) between 6 and 15 years old participated in this study. Handgrip strength was measured using a dynamometer, and the average value of both hands was used for analysis. Sports experience was determined when they started practicing each sport. Handgrip strength was compared between sports. Statistical moderation was used to determine if the relationship between sport and handgrip strength depended upon the age of the athlete. Results: Kendo athletes had significantly higher handgrip strength than soccer athletes (4.77 kg [95% CI: 2.34, 7.19]) in the overall sample. We found that the relationship between sport and handgrip strength depended upon the age of the child (sport*age t = −3.6, p =.004). Using the Johnson‐Neyman procedure, we found statistically significant differences between sports from 8.48 years and older. Conclusions: Our results suggest that the type of sport played, that is, whether or not an athlete plays with sports equipment in their hands, may influence the development of handgrip strength during the period of growth, and these sports may contribute to a higher level of handgrip strength in adulthood. [ABSTRACT FROM AUTHOR]

Free play in kindergarten can be roughly divided into fine and gross motor activities, but the effects of these activities on improving handgrip strength are unknown. Therefore, we aimed to compare one-year changes in handgrip strength and forearm flexor muscle size in children separated by preferred play in a kindergarten. One hundred and eleven children were recruited from a local kindergarten. They underwent handgrip strength and forearm muscle thickness measurements, and 95 (49 boys and 46 girls) underwent a second measurement one year after the first measurement. Class teachers assessed the physical activity of everyone in their class after the second measurement. Using three evaluation scores by the class teachers, we divided children into three groups based on the children's preference to play in kindergarten (fine movement vs. gross motor movement). Handgrip strength did not change differently between groups across one year. However, children who liked active playing outside (i.e., gross motor activity) were stronger than others. Furthermore, children who like playing outside observed greater changes than the other groups in the ulna (right hand) and radius muscle thickness (left hand), suggesting that changes in forearm muscle size might be incongruent with changes in handgrip strength among the three activity groups. [ABSTRACT FROM AUTHOR]

Objectives: The factors involved in changes in grip strength (GS) during growth/development are not well known. Findings from cross‐sectional studies have indicated that digit lengths are associated with physical fitness, including GS. This study aimed to investigate the association of changes in GS over 1 year and the second (2D) and fourth (4D) digit lengths in young children using the 4D as a covariate. Methods: One hundred and three young children (54 boys and 49 girls) performed maximum voluntary GS and ultrasound‐measured forearm muscle thickness measurements in the right hand. All participants completed the first measurement and underwent a second measurement 1 year after the first one. The 2D and 4D were taken on the palmar surface of the outstretched right hand at the second measurement. Results: The 2D was inversely associated with the change in GS (B = −2.1, p = 0.023) adjusted for all covariates. Girls had numerically lower adjusted changes in grip strength, although this was not statistically significant [−0.61 (−1.2, 0.02) kg]. When sex was removed from the model, the 2D remained inversely associated with the change in GS (B = −2.39, p = 0.011). Finally, when only adjusting for the 4D, the 2D was inversely associated with the change in GS (B = −3.07, p = 0.004). Conclusion: This study documented the association between changes in GS over 1 year and digit lengths in young children. The difference in children's digit length needs to be recognized as a factor involved in weak GS in children. [ABSTRACT FROM AUTHOR]

Objective: This study developed a scale for public health nurses supporting resident groups toward community‐building and verified its reliability and validity. Design and Sample: In this cross‐sectional investigation, self‐administered questionnaires were distributed to 1,924 public health nurses in Japanese municipalities. Measurements: The questionnaire included items on the public health nurses' demographic attributes, the developmental stage of the resident groups they supported, a draft scale, and an external criterion. Reliability was verified by calculating the alpha coefficient and test‐retest reliability. To clarify construct validity, we conducted exploratory and confirmatory factor analyses. Results: We analyzed 570 questionnaires. The scale was structured using 30 items covering four factors: "Creating opportunities to connect with community residents, other groups, and governments," "Stimulating activities by improving organizational capacity," "Promoting stable organizational management," and "Providing opportunities to review community health issues and activities." The Cronbach's alpha coefficient for this scale was 0.944. The intraclass correlation coefficient was 0.923 using test‐retest relativity. Correlations were noted for criterion‐related validity (r = 0.388, p <.01). Confirmatory factor analysis with structural equation modeling revealed a reasonable fit to the data. Conclusions: The scale for public health nurses supporting resident groups toward community‐building was confirmed to be reliable and valid. [ABSTRACT FROM AUTHOR]

Objective: A Smedley hand dynamometer is one of the standard devices for measuring handgrip strength (HGS) for children and adults. The aim was to compare the HGS values using two different types of dynamometers (Grip‐A or Grip‐D) in young children. To enable comparison between the two devices, we have redesigned the Grip‐D (i.e., modified Grip‐D). Methods: Twenty‐five preschool children (10 girls and 15 boys) performed maximal voluntary HGS in the right hand using two different types of dynamometers. We ran a paired sample t‐test on the difference in HGS between the two devices. Results: The measured values of HGS were 9.95 kg for Grip‐A and 8.56 kg for modified Grip‐D, and the difference between the two devices [1.39 (SD 0.65) kg] was greater than we expected (95% limits of agreement: 0.11, 2.6 kg). Thus, we then calibrated both dynamometers ourselves using known weights. The measured values were corrected if there was an error between the known weight and each dynamometer. Following adjustment, there was still a statistical difference (p < 0.001) in HGS between Grip‐A [10.65 (SD 1.52) kg] and modified Grip‐D [9.98 (SD 1.85) kg]. However, the difference between the two devices was 0.67 (SD 0.69) kg with the 95% limits of agreement between −0.68 and 2.0 kg. Conclusion: It is concluded that the HGS values of children measured with the company‐calibrated new Grip‐A and modified Grip‐D could provide reasonably close estimates. [ABSTRACT FROM AUTHOR]

Background: Contraction of bronchial smooth muscle is the main mechanism of asthmatic responses. Inflammatory cells such as eosinophils and mast cells produce chemical mediators that induce smooth muscle contraction. To investigate the mechanisms of IgE-independent asthmatic response in murine asthma models, a novel in vitro assay system using primary cultured mouse bronchial smooth muscle cells (BSMC) was explored. Method: Trachea and bronchi were taken from young mice and underwent primary culture. BSMC were expanded in culture and then embedded in a collagen gel. The well-known constrictors leukotriene D4 (LTD4), methacholine and histamine were applied to the BSMC gels. The gel images were captured by an image analyzer and contractile responses were evaluated. Results: LTD4 and methacholine significantly induced the gel contraction in a dose-dependent manner, but histamine did not. Montelukast, a CysLT type ΙΙ receptor antagonist, and atropine, a muscarinic acetylcholine receptor antagonist, inhibited the contractile responses in an agonist-specific manner. Conclusion: A contraction assay system using cultured mouse BSMC was successfully established for the first time. It may go a long way toward identifying bronchoconstricting mediators involved in murine asthma models. Copyright © 2013 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Background: Adoptive transfer of helper T (Th) cells conferred a late asthmatic response upon antigen challenge. A possible production of a contractile activity for bronchial smooth muscle (BSM) by activated Th cells was examined. Method: Murine Th clones were stimulated with immobilized anti-CD3 antibody in the presence or absence of anti-CD28 antibody. Culture supernatants were dialyzed and then applied to the collagen gels containing cultured human BSM cells. Results: Culture supernatants of activated but not resting murine Th clones that conferred a late asthmatic response, induced the contraction of human BSM cell-containing collagen gels. Conclusion: Activated Th cells produce a contractile activity for BSM in vitro. Copyright © 2012 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Background: Glucocorticoid (GC) action on asthma has been partly explained by the inhibition of T cell activation. We analyzed the steroid sensitivity of ovalbumin (OVA) reactive helper T (Th) cell clones both in vitro and in vivo. Method: For in vitro experiments, Th clones were cultured with antigen-presenting cells, OVA, and various concentrations of dexamethasone (DEX). The proliferative response of each Th clone was measured by 3H-thymidine uptake. For in vivo experiments, unprimed BALB/c mice were transferred with Th clones, challenged with OVA, and administered DEX subcutaneously. The number of infiltrating cells in bronchoalveolar lavage fluid (BALF) was measured. Results: Six Th clones were classified as steroid-sensitive or steroid-resistant clones in terms of the effects of GC on the proliferative responses analyzedin vitro. Airway infiltration of eosinophils and lymphocytes of mice transferred with steroid-sensitive clones were effectively inhibited by the administration of DEX. In contrast, those of mice transferred with steroid-resistant clones were not significantly inhibited by DEX; the number of eosinophils in the BALF of mice transferred with 1 steroid-resistant clone, i.e. T5-1, was only partially reduced. Conclusion: The steroid sensitivity of Th clones measured in vitro was consistent with that of an adoptively transferred asthma model measuredin vivo. Steroid-sensitive and resistant asthma models seem valuable for understanding the mechanisms of steroid resistance in severe asthma. Copyright © 2011 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Background: Helper T (Th) cells are deeply involved in the pathophysiology of bronchial asthma, such as eosinophilic inflammation, bronchial hyperresponsiveness (BHR), airflow limitation and remodeling. It is still unclear whether Th cells contribute to BHR independently of eosinophilic inflammation. The double GATA (dblGATA) site is a high-affinity GATA-binding site in the GATA-1 promoter. dblGATA site-deficient (ΔdblGATA) mice lack eosinophils. Method: Ovalbumin (OVA)-reactive Th clones were transferred into ΔdblGATA and wild-type (WT) mice of BALB/c background. The number of eosinophils in the bronchoalveolar lavage fluid (BALF) and bronchial responsiveness to methacholine were examined after OVA challenge. Results: The number of BALF eosinophils was significantly increased in WT mice, but not detectable in ΔdblGATA mice. BHR was also induced in WT mice, but significantly attenuated in ΔdblGATA mice. Conclusion: Eosinophils are involved in T-cell-mediated BHR. Copyright © 2010 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Fluoroalkyl end-capped N,N-dimethylacrylamide and acryloylmorpholine oligomers were polymerized with hydrolysis and condensation of tetraethoxysilane (TEOS) to afford homogeneous fluoroalkyl end-capped oligomers and silica gel polymer hybrids, respectively. In particular, the sol solutions of fluorinated oligomers/TEOS polymer hybrids were applied to surface modifications of glass with not only a strong oleophobicity but also good hydrophilicity. [ABSTRACT FROM AUTHOR]

The X-ray crystal structures of Thermoactinomyces vulgaris R-47 α-amylase 1 (TVAI) and α-amylase 2 (TVAII) have been determined at 1.6 A˚ and 2.3 A˚ resolution, respectively. The structures of TVAI and TVAII have been refined, R-factor of 0.182 (Rfree=0.206) and 0.179 (0.224), respectively, with good chemical geometries. Both TVAI and TVAII have four domains, N, A, B and C, and all very similar in structure. However, there are some differences in the structures between them. Domain N of TVAI interacts strongly with domains A and B, giving a spherical shape structure to the enzyme, while domain N of TVAII is isolated from the other domains, which leads to the formation of a dimer. TVAI has three bound Ca ions, whereas TVAII has only one. TVAI has eight extra loops compared to TVAII, while TVAII has two extra loops compared to TVAI. TVAI can hydrolyze substrates more efficiently than TVAII with a high molecular mass such as starch, while TVAII is much more active against cyclodextrins than TVAI and other α-amylases. A structural comparison of the active sites has clearly revealed this difference in substrate specificity. [Copyright &y& Elsevier]

We investigated the inhibitory effects of a non-acylguanidine Na... -H[sub+] exchange (NHE) inhibitor, T-162559 ((5E7SH7-{5-fluoro-2-methylphenyl)-4guanidine dimethanesulphonate), on NHE-1, and its cardioprotective effect against ischaemia and reperfusion injury in rats and rabbits. 2 T-162559 inhibited human platelet NHE-1 in a concentration-dependent manner, with an IC[sub50] value of 13 ±3 nmol l[sup-1], making it 16 and three times more potent than cariporide IC[sup50]: 209 ±75 nmol l[sup-1], P <0.01) and eniporide (IC[sub50]: 40 ±11 nmol l[sup-1], P=0.066), respectively. T-162559 also inhibited rat NHE-1 with an IC[sub50] value of 14 ±2 nmol 1[sup-1], which was five and three times lower than that of cariporide (IC[sub50]: 75 ±7 nmol l[sup-1], P <0.01) and eniporide (IC[sub50]: 44 ±2 nmot 1[sup-1], P<0.01), respectively. 3 T-162559 inhibited, in a concentration-dependent manner, the reduction in cardiac contractility, progression of cardiac contracture, and increase in lactate dehydrogenase release after global ischaemia and reperfusion in perfused rat hearts. The inhibitory effects of T-162559 were observed at a lower concentration range (10- 100 nmol 1[sup-1]) than with cariporide and eniporide, T-162559 did not alter basal cardiac contractility or coronary flow after reperfusion, suggesting that it exerts direct cardioprotective effects on the heart. 4 Intravenous administration of T-162559 (0.03 and 0.l mg kg[sup-1]) significantly inhibited the progression of myocardial infarction induced by left coronary artery occlusion and reperfusion in rabbits; the infarct size normalized by area at risk was 74 ±6% in the vehicle group, and 47 ±5% and 51 ±7% in the T-162559-0.03 mg kg[sup-1] and T-162559-0.1 mg kg[sup-1] groups (both P <0.05), respectively. 5 These results indicate that the new structural NHE-1 inhibitor T-162559 is more potent than cariporide and eniporide and possesses a cardioprotective effect against ischaemia and reperfusion injury in rat and rabbit models. [ABSTRACT FROM AUTHOR]

The influence of sediment contaminants on the Salmonella mutagenicity test was investigated to develop a preparation method not subject to interference by coexisting substances. Organic chemicals were extracted from sediment by dichloromethane/ethanol (4:1), acetonitrile or ethanol, and dichloromethane (DCM) extract was refined by washing with alkali, passing through a reduced-Cu column and/or fractionation with a silica cartridge. The strain Salmonella typhimurium TA98 was used. Sediment extracts and their refined preparations were first determined in their own mutagenicity, and then the influence on the mutagenicity of a fixed dose of 2-aminofluorene (2-AF) as a positive standard mutagen was examined. The toxicity to the strain was also tested. It was concluded that the coexisting environmental chemicals in sediment inhibited or promoted the original mutagenicity of the samples. [ABSTRACT FROM AUTHOR]

Asthma therapy in general has improved a lot in recent years, but it is still a major problem that severe asthma, which accounts for 10 to 20%, still suffers from strong symptoms on a daily basis despite all therapeutic agents used in combination. American SARP and European ENFUMOSA started in 2000 to advance pathophysiological insights of severe asthma. Clinical usage of antibodies and inhibitors against IgE, TNF, IL-5, IL-4, IL-13, and TSLP are also accumulating. Some of these molecular-targeted drugs improve respiratory function and reduce acute exacerbations in patients with severe asthma. Until now, cytokines have been assumed to be involved in chronic inflammation, but it is also interesting to elucidate the pathways of how cytokines are involved in respiratory function and acute exacerbations. We registered approximately 100 steroid-dependent asthma patients in Japan. Although long-lasting poor control of the disease was considered the cause of severe asthma in the past, steroid dependence in one third of the cases occurred within 2-3 years after the onset. Steroid resistance seems a key process from the early stage of the disease. Steroid resistance of T cell level was induced by extracellular co-stimulation and cytokine signals. The inhibition may improve steroid sensitivity and treat steroid-resistant asthma. Therefore, we established a steroid-resistant asthma model for the first time by transferring steroid resistant T cell clones, and analyzed the steroid sensitivity recovery effect of CTLA4-Ig. In addition, a multicenter, double-blind, placebo-controlled exploratory trial was performed as a POC study investigating the efficacy of abatacept in treatment-resistant severe asthma. Elucidation of the pathophysiology and mechanism by which steroids do not work is expected to be a breakthrough for the prevention and treatment of severe asthma.

Objective: To create a "Health Promotion Checklist for Residents" to help promote healthy habits among local residents., Methods: First, we investigated items for a health promotion checklist in the Health Japan 21 (2(nd) edition) and other references. Next, we conducted a questionnaire survey including these checklist items in August 2012. The study subjects were randomly selected Hatsukaichi city residents aged ≥20 years. Anonymous survey forms explaining this study were mailed to the investigated subjects and recovered in return envelopes. Data were compared by sex and age group., Results: We created a checklist comprising a 23-item health promotion evaluation index with established scoring. There were 33 questions regarding health checkups; cancer screenings; dental checkups, blood pressure; glycated hemoglobin or blood glucose; dyslipidemia; body mass index; number of remaining teeth; breakfast, vegetable, fruit, and salt intake; nutrient balance; exercise; smoking; drinking; sleep; stress; and mental state. There were also questions on outings, community involvement, activities to improve health, and community connections. The questions were classified into six categories: health management, physical health, dietary and exercise habits, indulgences, mental health, and social activities. Of the 4,002 distributed survey forms, 1,719 valid responses were returned (recovery rate, 43.0%). The mean score by category was 1.69 (N=1,343) for health management, 6.52 (N=1,444) for physical health, 12.97 (N=1,511) for dietary and exercise habits, and 2.29 (N=1,518) for indulgences, all of which were higher for women, and 5.81 (N=1,469) for mental health, which was higher for men. The health management scores were higher among subjects in their 40s and 50s. The physical health score increased gradually with age from the 70 s and older to the 20 s, whereas the dietary and exercise habits increased gradually from the 20 s to the 70 s and older. The 20 s had high scores for indulgences, while mental health was low for the 20 s and 30 s and gradually increased from the 40 s to the 70 s and older. The social activities score (1.93; N=1,539) tended to be higher in the 40 s and older., Conclusion: Here we created and attempted to validate a checklist that promotes healthy habits nd found that subjects were able to use it to examine their living habits.

Acyl-CoA thioesterases (ACOTs) are a group of enzymes that catalyze the hydrolysis of fatty acyl-CoAs to free fatty acids and CoA, with the potential to regulate the intracellular levels of these molecules. In this study, we show that a cytosolic isoform, ACOT7, is expressed at a significant level in the mesenteric lymph nodes (MLNs) of mice. While crude preparations of the mesenteric visceral fat contained significant levels of palmitoyl-CoA thioesterase activity, enzyme activity was concentrated 6.9-fold in MLNs compared with the residual adipose portion after excision of MLNs. When MLN homogenates were centrifuged, 82% of the enzyme activity was recovered in the cytosolic fraction, concomitant with almost exclusive recovery of ACOT7. Immunoprecipitation using anti-ACOT7 antibody estimated that 87% of enzyme activity in the homogenates was accounted for by ACOT7. On MLN sections, the germinal centers of secondary lymphoid follicles were immunostained with the antibody. In MLNs of mice fasted for 16 h, ACOT7 levels were induced 1.8-fold, which reflected a 1.5-fold increase in enzyme activity. These findings suggest that ACOT7 may be involved in dietary intake-associated responses in fatty acid metabolism in MLNs.

Thermoactinomyces vulgaris R-47 alpha-amylase 1 (TVAI) has unique hydrolyzing activities for pullulan with sequence repeats of alpha-(1,4), alpha-(1,4), and alpha-(1,6) glycosidic linkages, as well as for starch. TVAI mainly hydrolyzes alpha-(1,4) glycosidic linkages to produce a panose, but it also hydrolyzes alpha-(1,6) glycosidic linkages with a lesser efficiency. X-ray structures of three complexes comprising an inactive mutant TVAI (D356N or D356N/E396Q) and a pullulan model oligosaccharide (P2; [Glc-alpha-(1,6)-Glc-alpha-(1,4)-Glc-alpha-(1,4)]2 or P5; [Glc-alpha-(1,6)-Glc-alpha-(1,4)-Glc-alpha-(1,4)]5) were determined. The complex D356N/P2 is a mimic of the enzyme/product complex in the main catalytic reaction of TVAI, and a structural comparison with Aspergillus oryzaealpha-amylase showed that the (-) subsites of TVAI are responsible for recognizing both starch and pullulan. D356N/E396Q/P2 and D356N/E396Q/P5 provided models of the enzyme/substrate complex recognizing the alpha-(1,6) glycosidic linkage at the hydrolyzing site. They showed that only subsites -1 and -2 at the nonreducing end of TVAI are effective in the hydrolysis of alpha-(1,6) glycosidic linkages, leading to weak interactions between substrates and the enzyme. Domain N of TVAI is a starch-binding domain acting as an anchor in the catalytic reaction of the enzyme. In this study, additional substrates were also found to bind to domain N, suggesting that domain N also functions as a pullulan-binding domain.

The X-ray structures of complexes of Thermoactinomyces vulgaris R-47 alpha-amylase 1 (TVAI) with an inhibitor acarbose and an inactive mutant TVAI with malto-hexaose and malto-tridecaose have been determined at 2.6, 2.0 and 1.8A resolution, and the structures have been refined to R-factors of 0.185 (R(free)=0.225), 0.184 (0.217) and 0.164 (0.200), respectively, with good chemical geometries. Acarbose binds to the catalytic site of TVAI, and interactions between acarbose and the enzyme are very similar to those found in other structure-solved alpha-amylase/acarbose complexes, supporting the proposed catalytic mechanism. Based on the structure of the TVAI/acarbose complex, the binding mode of pullulan containing alpha-(1,6) glucoside linkages could be deduced. Due to the structural difference caused by the replaced amino acid residue (Gln396 for Glu) in the catalytic site, malto-hexaose and malto-tridecaose partially bind to the catalytic site, giving a mimic of the enzyme/product complex. Besides the catalytic site, four sugar-binding sites on the molecular surface are found in these X-ray structures. Two sugar-binding sites in domain N hold the oligosaccharides with a regular helical structure of amylose, which suggests that the domain N is a starch-binding domain acting as an anchor to starch in the catalytic reaction of the enzyme. An assay of hydrolyzing activity for the raw starches confirmed that TVAI can efficiently hydrolyze raw starch.