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Eiichi Saitoh

Publications and source records attributed to Eiichi Saitoh.

17 recordsLinked to original sources
PubMed2006

Extracellular production of human cystatin S and cystatin SA by Bacillus subtilis.

We herein describe the development of a Bacillus subtilis system that can be used to produce large quantities of recombinant (r-) human salivary cystatins, a cysteine protease inhibitor of family 2 in the cystatin superfamily. The B. subtilis that lacked the alkaline protease E gene (DeltaaprE type mutant strain) was prepared by homologous recombination. The cDNA fragments coding for mature cystatins (S and SA) were ligated in frame to the DNA segment for the signal peptide of endoglucanase in the pHSP-US plasmid vector that was then use to transform the DeltaaprE type mutant strain of B. subtilis. The transformants carrying the expression vectors were cultivated in 5-L jar fermenters for 3 days at 30 degrees C. Both r-cystatin S and r-cystatin SA were successfully expressed and secreted into the culture broth, and were purified using a fast performance liquid chromatography system. The first use of DeltaaprE type mutant strain of B. subtilis made it possible to obtain a high yield of secreted protein, which makes this system an improvement over expression in Escherichia coli. We conclude that this system has high utility for expression of commercial quantities of secreted proteins.

Bacillus subtilis↗
PubMed2005

Molecular cloning and characterization of oryzacystatin-III, a novel member of phytocystatin in rice (Oryza sativa L. japonica).

On the basis of cDNA sequences, we found that the calli of rice encodes an amino acid sequence that shares 56% and 89% identity, respectively, with oryzacystatin-I and oryzacystatin-II. This sequence differs from that of oryzacystatin-II in the N-terminal region (Gln(7)-Ala(19) in the oryzacystatin-III numbering), and this region contained a glycine residue (Gly(14)), which is evolutionarily conserved in the cystatin superfamily. We named this novel protein oryzacystatin-III. Nucleotide sequencing of the 5'-flanking region of the oryzacystatin-III gene showed that it is highly homologous to the oryzacystatin-II gene but distinct from the oryzacystatin-II locus. Oryzacystatin-III inhibited papain, ficin, and human cathepsin B. The inhibition constants for papain and ficin differ from those of oryzacystatin-I and -II, and cathepsin B activity is affected only by oryzacystatin-III, showing differences in the interaction of these inhibitors with enzymes. These data suggest that the above three inhibitors may play unique physiological roles in the regulations of rice cysteine proteinases.

Adaptor Proteins, Signal Transducing↗
PubMed2005

Murine monoclonal antibody which can distinguish cystatins SA1 and SA2.

To develop a diagnostic trial enabling the selective examination for a target cystatin in human body fluids, we attempted to prepare monoclonal antibodies against human cystatin SA1 (originally cystatin SA) and its variant form (cystatin SA2). BALB/c mice were immunized with recombinant (r-) cystatins SA1 and SA2. Two monoclonal antibodies designated Cys3F11 and Cys2E5 were selected. By ELISA analyses, the Cys2E5 was shown to react with r-cystatin SA2 but also somewhat with r-cystatin SA1 (22% cross-reactivity) and with plasma cystatin C (18% cross-reactivity), indicating a high specificity for cystatin SA2. The Cys3F11 reacted not only with r-cystatin SA1 but also with r-cystatin SA2 (89% cross-reactivity) and plasma cystatin C (47% cross-reactivity). This finding was further emphasized by immunoblotting of human submandibular-sublingual saliva samples. ELISA additivity test suggests that the two monoclonal antibodies bind to distinct epitopes. In conclusion, we have succeeded in producing two antibodies that discriminate the structural differences between salivary cystatins S and SN, which share more than 90% identity in amino acid sequence with cystatin SA.

Amino Acid Sequence↗
PubMed2005

A novel cysteine protease inhibitor with lectin activity from the epidermis of the Japanese eel Anguilla japonica.

A novel cysteine protease inhibitor (Eel-CPI-1) was isolated from the epidermis of the eel. Eel-CPI-1 was shown to bind strongly to both lactose- and carboxymethylated papain-affinity gels. Its molecular mass under reducing condition was determined to be 18 kDa by SDS-polyacrylamide gel electrophoresis but approximately 30.5 kDa under non-reducing-conditions. Eel-CPI-1 inhibited papain (K(i)=18 nM) and ficin (K(i)=120 nM) competitively. Combined with the data on amino acid and sequence analysis, Eel-CPI-1 is identical to the eel lectin, AJL-2. This is the first report describing a cysteine protease inhibitor with lectin activity.

Animals↗
PubMed2005

Development of a novel type of shoe to improve the efficiency of knee-ankle-foot orthoses with a medial single hip joint (Primewalk orthoses): a novel type of shoe for Primewalk orthosis.

The purpose of the study was to develop and evaluate a new heel cushion in shoes for use with knee-ankle-foot orthoses having a medial single hip joint (Primewalk orthoses) in order to improve walking velocity and efficiency. Primewalk orthoses and shoes were made for a 24-year-old man having paraplegia with flaccid paresis (level T-7; grade A, ASIA impairment scale) of 2 years' duration. Walking exercises were assigned. Shoes were modified with the sole made of hard rubber and the addition of soft rubber heel cushions. The walking speed, centre of foot pressure during walking, and ground reaction force were evaluated. The patient also subjectively assessed the devices. The modifications to the shoes resulted in a 1.94-fold increase in walking speed (8.6 to 16.7 m/min), a 1.87-fold increase in step length (16.7 to 31.3 cm) and a 54.8% decrease in the physiological cost index (7.7 to 3.48 beats/min). The centre of foot pressure during walking was found to deviate towards the lateral margin of the foot. The horizontal rotation of the pelvis increased simultaneously. The patient reported increased amplitude of flail motion of the trunk and decreased burden to the upper limbs. It was concluded that the modified new heel cushion of the shoe provided freedom to the lower legs and thereby increased walking efficiency.

Adult↗
PubMed2005

Reliability and validity of a wheelchair collision test for screening behavioral assessment of unilateral neglect after stroke.

OBJECTIVE: To examine the reliability and validity of a new wheelchair collision test (WCT) for screening behavioral unilateral neglect in people with stroke. DESIGN: A total of 19 patients with left hemiplegia resulting from first-ever stroke (mean +/- SD, 61.9 +/- 25.8 days after onset) were examined with the WCT, whereby the patient was asked to propel a wheelchair to pass four round chairs arranged in two rows. Separate tests were done using two different distances between the chairs: 120 and 140 cm. The number of collisions with the chairs was recorded. The Catherine Bergego Scale was used to evaluate behavioral unilateral neglect. Because unilateral neglect has an unfavorable impact on rehabilitation, the FIMtrade mark instrument was used to determine the function of daily activities. RESULTS: Test-retest reliability ranged from 0.68 to 0.97 in different situations in the WCT. The rate of agreement tested by Kappa statistics between the WCT and Catherine Bergego Scale was higher when the distance between chairs was 120 cm than when the distance was 140 cm (0.68 vs. 0.58). Spearman's rank-correlation coefficient between the WCT and Catherine Bergego Scale was 0.72 (P < 0.001) if the distance between the chairs was 120 cm and 0.75 (P < 0.001) if the distance was 140 cm. The WCT and Catherine Bergego Scale correlated well with the FIM instrument. Spearman's rank-correlation coefficient between the Catherine Bergego Scale and FIM total score was -0.70 (P < 0.001). CONCLUSION: The WCT is a simple screening test with high test-retest reliability and validity to evaluate behavioral unilateral neglect.

Aged↗
PubMed2005

Line bisection and rebisection: the crossover effect of space location.

OBJECTIVE: To investigate the relationship between the bisection test and the severity of behavioral hemineglect and to verify if this test can predict the behavioral hemineglect. METHODS: Thirty stroke patients with left hemiparesis were divided into 4 groups according to the Catherine Bergego Scale, which assessed the behavioral hemineglect: severe unilateral neglect (UN), moderate UN, mild UN, and lack of UN. Eleven healthy subjects served as age-matched control subjects. In the bisection test, 18 lines were presented on the left, middle, and right of an A4 paper, respectively. The subjects were asked to place a short cross mark in the exact middle point of each line on the paper using their right hand. The middle 6 lines in the above bisection test were extracted on another sheet of A4 paper for the rebisection test. The subjects were asked to divide a line into 4 segments by successive bisections. The proportion of the right part to the length of line for bisecting was calculated. RESULTS: In the bisection test, the main effect of space was significant in every group except the mild neglect group. The crossover effect of space location was found in the severe UN group, the group without UN, and the controls. In the severe UN group, the patients bisected the left and middle lines with rightward bias (<50%) but bisected the right lines with leftward bias (>50%). In the group without UN and the controls, the subjects bisected the left lines with leftward bias (>50%) but bisected the middle and right lines with rightward bias (<50%). Almost the same results were seen in the rebisection test. CONCLUSIONS: This study showed that if the spatial crossover effect occurred in the right space condition, it was strongly supported that this patient had moderate to severe behavioral hemineglect. The crossover effect of the space location was explained by a new model.

Activities of Daily Living↗
PubMed2004

Full-time integrated treatment program, a new system for stroke rehabilitation in Japan: comparison with conventional rehabilitation.

OBJECTIVE: To validate the effectiveness of the Full-time Integrated Treatment (FIT) program that is characterized by rehabilitation 7 days/wk, encouragement of daytime activity, and enhanced communication between staff in stroke rehabilitation. DESIGN: Since our facility changed from the conventional rehabilitation system of 5 days of treatment to the FIT program in December 2000, we compared the conventional rehabilitation program with the FIT program at our hospital. The conventional treatment group and the FIT group consisted of 48 and 58 first-stroke hemiplegics, respectively. RESULTS: The motor subscore of the FIM instrument at admission and at discharge was 64.3 and 77.0 in the conventional group and 60.6 and 80.9 in the FIT group, respectively. The length of stay and efficiency of the FIM instrument were 80.0 days and 0.16 in the conventional group and 69.8 days and 0.30 in the FIT group, respectively. These differences between groups were statistically significant, with the exception of admission FIM data. CONCLUSIONS: Because the FIT program attained a higher discharge FIM level with a shorter length of stay, the FIT program was concluded to be an efficient and effective method of stroke rehabilitation.

Activities of Daily Living↗
PubMed2004

Cystatin SA, a cysteine proteinase inhibitor, induces interferon-gamma expression in CD4-positive T cells.

Recently, it has been demonstrated that family 2 cystatins upregulate interleukin-6 production by human gingival fibroblasts. In the present study, we investigated the effects of cystatin SA on cytokine production by helper T cells. Human CD4-positive T cells were cultured with phytohemagglutinin in the presence or absence of 0.1 microM recombinant cystatin SA1 or SA2. When the amounts of interleukin-4 (IL-4) and interferon-gamma (IFNgamma) were analyzed in an ELISA system after stimulation with either cystatin, no significantly increased levels of IL-4 were detected. However, the amounts of IFNgamma were significantly increased after stimulation with the cystatins. Our results suggest that salivary family 2 cystatins are involved in immune responses through the cytokine network.

CD4-Positive T-Lymphocytes↗
PubMed2003

Three tests for predicting aspiration without videofluorography.

The videofluorographic swallowing study (VFSS) is the definitive test to identify aspiration and other abnormalities of swallowing. When a VFSS is not feasible, nonvideofluorographic (non-VFG) clinical assessment of swallowing is essential. We studied the accuracy of three non-VFG tests for assessing risk of aspiration: (1) the water swallowing test (3 ml of water are placed under the tongue and the patient is asked to swallow); (2) the food test (4 g of pudding are placed on the dorsum of the tongue and the patient asked to swallow); and (3) the X-ray test (static radiographs of the pharynx are taken before and after swallowing liquid barium). Sixty-three individuals with dysphagia were each evaluated with the three non-VFG tests and a VFSS; 29 patients aspirated on the VFSS. The summed scores of all three non-VFG tests had a sensitivity of 90% for predicting aspiration and specificity of 71% for predicting its absence. The summed scores of the water and food tests (without X-ray) had a sensitivity of 90% and specificity of 56%. These non-VFG tests have limitations but may be useful for assessing patients when VFSS is not feasible. They may also be useful as screening procedures to determine which dysphagia patients need a VFSS.

Adolescent↗
PubMed2003

[The relationship between normal swallowing functions, food ingestion and the lifestyle of users in a day service center].

OBJECTIVES: The physical and mental condition of elderly people changes every day and we hypothesized that this influences food intake and swallowing functions. The present study was undertaken to clarify relationships between daily living conditions and swallowing function. METHODS: The subjects were users (105 males and 219 females) of 6 day-service centers. We performed a survey of IADL (Instrumental Activities of Daily Living), mobility, eating and swallowing behavior, and general health. In addition, we used RSST (Repetitive Saliva Swallowing Test) as a screen for functional dysphasia. RESULTS: Women who had normal food ingestion and swallowing functions were capable of doing shopping, housework and taking care of money independently. Males and females who were capable of moving around from place to place had normal food ingestion and swallowing functions. With regard to daily eating habits and health condition, males and females who had normal food ingestion and swallowing functions were able to consume ordinary food, able to eat without any help, and often laugh. CONCLUSION: It was concluded that a normal swallowing condition in the elderly is related to laughing often, being independent in daily life, moving around and eating ordinary meals without any help.

Activities of Daily Living↗
PubMed2002

Ontogenetic changes in neuropeptide Y-immunoreactive cerebrospinal fluid-contacting neurons in the hypothalamus of the cloudy dogfish, Scyliorhinus torazame (Elasmobranchii).

Ontogenetic changes in neuropeptide Y-immunoreactive (NPY-ir) cerebrospinal fluid (CSF)-contacting neurons in the dogfish hypothalamus were studied immunohistochemically. NPY-ir CSF-contacting neurons first appeared in the median infundibular floor of the embryo at the 34 mm stage. At the 40 mm stage, similar neurons were found also in the saccus vasculosus (SV). The number of these neurons increased during the 54-80 mm stages, and the cells in the infundibular floor extended their basal processes to the neuropil of the median eminence, whereas the cells in the SV sent their axonal fibers to the tractus sacci vasculosi. After hatching, NPY immunoreactivity in the ventral hypothalamus became less dense, and the labeled CSF-contacting neurons tended to be confined to the nucleus lateralis tuberis, similarly as in the adults. The occurrence of NPY-ir CSF-contacting neurons in the SV was transient during the embryonic periods.

Animals↗
PubMed2002

Functional Neuromuscular Stimulation for articular angle control with an Inverse Dynamics Model tuned by a neural network.

A Functional Neuromuscular Stimulation (FNS) system was developed to restore the motor function of people with central nervous system disorders. An FNS articular angle control system with an Inverse Dynamics Model (IDM) tuned by a neural network was proposed. The system was designed to control the elbow joint angles by learning the characteristics of the neuromuscular system of controlled limbs. Three kinds of control schemes using the IDM were proposed. In a simulation experiment, the system was able to learn the abnormal characteristics of a limb and any changes in these characteristics, and then allow for better control of the limb. The results of experiments using human participants showed that the system had ability to control both normal and paralysed limbs.

Biophysical Phenomena↗
PubMed2002

Salivary cystatins induce interleukin-6 expression via cell surface molecules in human gingival fibroblasts.

Recently, family 2 cystatins have been demonstrated to upregulate interleukin-6 (IL-6) production by human gingival fibroblasts [Biol. Chem. 381 (2000) 1143]. To elucidate the mechanism of the IL-6 inducing activity of cystatins, we tested NF-kappa B activation with salivary cystatins SA1 and SA2-stimulated human gingival fibroblast whole cell lysates. The IL-6 production by human fibroblasts in response to these cystatins was inhibited by tyrosine kinase inhibitors and an inhibitor of NF-kappa B activation. The IL-6 inducing activity of the cystatins was depressed by the anti-CD58 monoclonal antibody. These findings supply evidence that cystatins SA1 and SA2 adhere to human fibroblasts and that the event results in tyrosine phosphorylation and upregulation of the release of IL-6 mediated enhancement of NF-kappa B activity.

Cell Line↗
PubMed2002

Sac I Restriction Fragment Length Polymorphism (RFLP) related to the human CST2 gene.

Restriction Fragment Length Polymorphism (RFLP) related to cystatin gene (CST) family was detected in the Japanese population by using restriction enzyme Sac I. A polymorphic site, located at 0.9 kb from the 3' end of the CST2 gene, revealed a two allele polymorphism with band sizes of 3.5 kb and 8.3 kb by hybridization with probe including exon 2 of the CST1 gene. The gene frequencies in the Japanese population were 0.326 for 3.5 kb allele and 0.674 for 8.3 kb allele (n = 86). The phenotypes of the polymorphism showed no association with the previously reported electrophoretic cystatin SA protein phenotypes.

Alleles↗
PubMedDate not supplied

Incidence and consequence of falls in inpatient rehabilitation of stroke patients.

The incidence rate, location, and timing of falls and performance of activities of daily living (ADLs) in 256 patients with stroke admitted to an 88-bed rehabilitation ward was observed with information on falls and level of ADLs upon admission extracted from patients' records. Of 273 falls among 121 of the 256 patients, 229 occurred in the patient's room or lavatory and 147 within 4 weeks of admission. Significant differences were detected between motor subscores under 64 and over 65 on the Functional Independence Measure (FIM) in fall proportional analysis. The mean fall rate in motor subclass of 26 to 38 was higher than in other subgroups. Those with cognitive subscore on the FIM lower than 29 were prone to fall. Also, deteriorated motor and cognitive functions were associated with a high risk of falls.

Accidental Falls↗