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1.  Influence of the difference of breastfeeding volume on a rat model of oxygen-induced retinopathy 
This study aimed to investigate the effects of abundant breast milk intake on rats model of oxygen-induced retinopathy (OIR). Neonatal Sprague–Dawley rats were randomly assigned to expand litters of 7 pups/litter (7-rats group) and 14 pups/litter (14-rats group). They were exposed to 80% oxygen from postnatal day (P) 0 to P12. Body weights were measured daily. At P13 and 18, rats were sacrificed, and the blood and eyes were collected. Retinal neovascularization (NV) score, total retinal area (TRA), avascular area (AVA), and vascularized area (VA) were measured in ADPase stained retinas. Retinal vascular endothelial growth factor (VEGF) and serum insulin-like growth factor (IGF-1) were measured using ELISA. Body weight gain was significantly greater in 7-rats group from P2. Serum IGF-1 levels at P13 and 18 were significantly higher in 7-rats group. Retinal VEGF and TRA at P18 were significantly larger in 7-rats group. NV score at P18 tended to be higher in 7-rats group. There was no significant difference in VA between the 2 groups at P13 and 18. Excess breast milk intake in OIR rat pups caused body weight gain and retinal development, whereas there was less effect on retinal vascularization in our study.
PMCID: PMC4186380  PMID: 25320460
nutrition; retinopathy of prematurity; oxygen-induced retinopathy; vascular endothelial growth factor; insulin-like growth factor
2.  Relationship between lifestyle and lifestyle-related factors in a rural–urban population of Japan 
To clarify the actual state of residents’ lifestyle in a mixed rural–urban area in Japan, and to investigate the relationship between residents’ lifestyle and lifestyle-related factors.
The Japanese version of Health Promoting Lifestyle Profile-II (HPLP-II), lifestyle-related factors developed through group work with residents of Town A, and demographic variables were used to evaluate 1176 community residents’ lifestyles and associated factors.
Factor analysis revealed that there were 4 factors related to healthy lifestyle. Nonparametric analysis revealed that female and elderly groups showed higher overall HPLP-II score than male and young groups. A significant correlation coefficient was seen between scores of overall HPLP-II and lifestyle-related factors (r = 0.611, p < 0.001). Multiple linear regression analysis demonstrated that HPLP-II was significantly associated with each lifestyle-related factor, showing a similar order in both gender and age groups. Finally, covariance structure analysis demonstrated that the score of health cognition and regional factors increased the score of HPLP-II, which then increased the score of self-rated health.
The present research clarified the actual state of residents’ lifestyles by age and gender in a mixed rural–urban area in Japan, demonstrating a vector model from health cognition and regional factors to self-rated health, via residents’ lifestyle.
PMCID: PMC3709039  PMID: 23160850
Quality of life; Self-rated health; HPLP-II; Lifestyle-related factors
4.  Community factors to promote parents’ quality of child-nurturing life 
The purpose of this study was to clarify the role of community factors in parents’ quality of child-nurturing life (QCNL).
We developed a questionnaire to evaluate the degree of QCNL and determine the structural factors related to QCNL as community factors related to parents’ QCNL derived from focus group interviews and the Delphi technique. The questionnaire also included the battery of the self-rating depression scale and Tsumori-Inage Infant's Developmental Test. Using the questionnaire, we then conducted a quantitative survey of parents whose children attended nursery schools in Kumamoto Prefecture. Factor analysis, calculation of the mean score and/or ratio to each item, Pearson’s correlation coefficient, t test, multiple regression analysis, and covariance structure analysis were performed.
The questionnaire we developed consisted of seven items with 75 elements, involving ten elements as community factors. Subjects included 699 parents (mean age 33.6 ± 5.4 years) and 965 children (age range 0–6 years). Factor analysis revealed that community factors consisted of five factors, such as “lifestyle rooted in the ground,” “balance of housekeeping and work,” “community network,” “amenity,” and “regeneration of life”. These factors may be dominant in a rural area. Finally, we developed a structural model with “community factors,” QCNL, QOL, and “child growth” by covariance structural analysis. The analysis revealed that community factors had a positive relation to parents’ QCNL (r = 0.81, p < 0.001) and that parental SDS score had a negative relation to parents’ QCNL (r = −0.59, p < 0.001). The analysis did show that community factors were positively related to the sound growth of children.
The covariance structure analysis revealed that community factors were associated with parents’ QCNL, SDS, and “child growth.”
PMCID: PMC3541813  PMID: 23054992
Maternal and child health; Quality of child-nursing life; Qualitative study; Quantitative study
5.  Pegylated Interferon-α2a Inhibits Proliferation of Human Liver Cancer Cells In Vitro and In Vivo 
PLoS ONE  2013;8(12):e83195.
We investigated the effects of pegylated interferon-α2a (PEG-IFN-α2a) on the growth of human liver cancer cells.
The effect of PEG-IFN-α2a on the proliferation of 13 liver cancer cell lines was investigated in vitro. Cells were cultured with medium containing 0–4,194 ng/mL of PEG-IFN-α2a, and after 1, 2, 3, or 4 days of culture, morphologic observation and growth assay were performed. After hepatocellular carcinoma (HCC) cells (HAK-1B and KIM-1) were transplanted into nude mice, various doses of PEG-IFN-α2a were subcutaneously administered to the mice once a week for 2 weeks, and tumor volume, weight, and histology were examined.
PEG-IFN-α2a inhibited the growth of 8 and 11 cell lines in a time- and dose-dependent manner, respectively, although the 50% growth inhibitory concentrations of 7 measurable cell lines on Day 4 were relatively high and ranged from 253 ng/mL to 4,431 ng/mL. Various levels of apoptosis induction were confirmed in 8 cell lines. PEG-IFN-α2a induced a dose-dependent decrease in tumor volume and weight, and a significant increase of apoptotic cells in the tumor. Subcutaneous administration of clinical dose for chronic hepatitis C (3 μg/kg, 0.06 μg/mouse) was effective and induced about 30-50% reduction in the tumor volume and weight as compared with the control.
Although in vitro anti-proliferative effects of PEG-IFN-α2a were relatively weak, PEG-IFN-α2a induced strong anti-tumor effects on HCC cells in vivo. The data suggest potential clinical application of PEG-IFN-α2a for the prevention and treatment of HCC.
PMCID: PMC3861497  PMID: 24349459
6.  Aldehyde Dehydrogenase 1 Identifies Cells with Cancer Stem Cell-Like Properties in a Human Renal Cell Carcinoma Cell Line 
PLoS ONE  2013;8(10):e75463.
Cancer stem cells (CSC) or cancer stem cell-like cells (CSC-LCs) have been identified in many malignant tumors. CSCs are proposed to be related with drug resistance, tumor recurrence, and metastasis and are considered as a new target for cancer treatment; however, there are only a few reports on CSCs or CSC-LCs in renal cell carcinoma (RCC). Different approaches have been reported for CSC identification, but there are no universal markers for CSC. We used two different approaches, the traditional side population (SP) approach, and the enzymatic (aldehyde dehydrogenase 1 (ALDH1)) approach to identify CSC-LC population in two RCC cell lines, ACHN and KRC/Y. We found that ACHN and KRC/Y contain 1.4% and 1.7% SP cells, respectively. ACHN SP cells showed a higher sphere forming ability, drug resistance, and a slightly higher tumorigenic ability in NOD/SCID mice than Non-SP (NSP) cells, suggesting that cells with CSC-LC properties are included in ACHN SP cells. KRC/Y SP and NSP cells showed no difference in such properties. ALDH1 activity analysis revealed that ACHN SP cells expressed a higher level of activity than NSP cells (SP vs. NSP: 32.7% vs 14.6%). Analysis of ALDH1-positive ACHN cells revealed that they have a higher sphere forming ability, self-renewal ability, tumorigenicity and express higher mRNA levels of CSC-LC property-related genes (e.g., ABC transporter genes, self-replication genes, anti-apoptosis genes, and so forth) than ALDH1-negative cells. Drug treatment or exposure to hypoxic condition induced a 2- to 3-fold increase in number of ALDH1-positive cells. In conclusion, the results suggest that the ALDH1-positive cell population rather than SP cells show CSC-LC properties in a RCC cell line, ACHN.
PMCID: PMC3792959  PMID: 24116047
7.  Symptomatic Middle Cerebral Artery Stenosis Treated by Percutaneous Transluminal Angioplasty: Improvement of Cerebrovascular Reserves 
Interventional Neuroradiology  2012;18(2):213-220.
This study evaluated the recoveries of cerebrovascular reserves (CVR) after applying percutaneous transluminal angioplasty (PTA) to patients with symptomatic middle cerebral artery (MCA) stenosis of varying severity. The patients were submitted to single photon emission computed tomography (SPECT) to obtain their regional cerebral blood flows at resting stage (rCBFrest) and acetazolamide-challenged CBF in five regions of interest (ROIs), including the MCA, on the ipsilateral and contralateral sides of the hemisphere. rCVR values were then calculated from these CBF data to evaluate the CVR recoveries after PTA treatment. When the PTA effects were statistically analyzed of the patients dichotomized into more severe (n=9) and less severe (n=5) groups, distinctly significant ROI-specific PTA effectiveness was observed for CVR rather than CBF values in the patients of the severer group.
PMCID: PMC3380402  PMID: 22681739
cerebrovascular reserves, angioplasty, middle cerebral artery stenosis, cerebral blood flow, ischemic stroke
8.  Assessment of health-promoting lifestyle profile in Japanese university students 
This descriptive study of 314 students was conducted to determine whether there were any relationships of their demographic variables with a health-promoting lifestyle profile.
The Japanese version of the Health Promoting Lifestyle Profile II (HPLP-II) questionnaire was used to investigate students’ lifestyles. One-way analysis of variance (ANOVA) and multiple comparison procedures using the Scheffe test were conducted to identify significant differences among university year (first, second, etc.) groups. The t-test independent participants was utilized to compare the HPLP-II scores according to gender and living with family (yes/no). Multiple regression analysis was used to analyze the effects of various demographics on the overall HPLP-II score and the six health-promoting lifestyle subscales.
The whole cohort of participants had an overall HPLP-II mean score of 2.50 (SD = 0.29), with the highest mean score being for interpersonal relations (3.05 ± 0.44), and the lowest mean score being for health responsibility (2.01 ± 0.53). The overall HPLP-II score of the students during the first university year was 2.59 (SD = 0.29), and this score progressively declined in their second year (2.49 ± 0.29), third year (2.47 ± 0.28), and fourth year (2.45 ± 0.30). Female students practiced significantly better health responsibility, interpersonal relations, and nutrition than males, and male students practiced significantly better physical activity than females. The students who were living with family had significantly higher nutrition scores than those not living with family.
These significant findings linking the HPLP-II, university year level, and living with family (yes/no) in university students will enable healthcare providers to develop interventions to assist students in improving their health lifestyles in the university environment and will help in devising suitable education programs.
PMCID: PMC3348247  PMID: 21987366
Health-promoting lifestyle profile II; Health behaviors; University year (first, second, etc.); University students; Lifestyle
9.  Transmesocolic Hernia of the Ascending Colon with Intestinal Obstruction 
Case Reports in Gastroenterology  2012;6(2):344-349.
An internal hernia may be either congenital or acquired. The reported incidence of such hernias is 1–2%. In rare cases, internal hernias are the cause of small bowel obstruction, with a reported incidence of 0.2–0.9%. Transmesocolic hernia of the ascending colon is especially rare. We report a case of transmesocolic hernia of the ascending colon with intestinal obstruction diagnosed preoperatively. A 91-year-old Japanese female was admitted to our hospital with abdominal distention and vomiting of 3 days duration. She had no past history of any abdominal surgery. Abdominal examination revealed distention and tenderness in the right iliac fossa. Abdominal computed tomography revealed ileus in the sac at the left side of the ascending colon and dilatation of the oral side of the intestine. We diagnosed a transmesocolic hernia of the ascending colon with intestinal obstruction and performed emergency surgery. At the time of operation, there was internal herniation of ileal loops through a defect in the ascending mesocolon, without any strangulation of the small bowel. The contents were reduced and the tear in the ascending mesocolon was closed. The postoperative course was uneventful and the patient was discharged 14 days after surgery. In conclusion, preoperative diagnosis of bowel obstruction caused by a congenital mesocolic hernia remains difficult despite the techniques currently available, so it is important to consider the possibility of a transmesocolic hernia when diagnosing a patient with ileus with no past history of abdominal surgery.
PMCID: PMC3383250  PMID: 22740809
Transmesocolic hernia; Ascending colon; Intestinal obstruction
10.  A comparative study of health-promoting lifestyles in agricultural and non-agricultural workers in Japan 
To clarify the difference in health-promoting lifestyles between agricultural and non-agricultural workers in Japan, a cross-sectional study was conducted on 627 residents living in a town with a mixed rural–urban population.
The subjects were divided into 8 groups by job (agricultural and non-agricultural), age (young and old), and gender (male and female). To evaluate the subjects’ lifestyles, the Health Promoting Lifestyle Profile II (HPLP-II) was applied. The Bartlett test and the Kendall rank test were performed for statistical analysis.
There was no significant difference in the overall score of the HPLP-II between the two job groups. However, for the HPLP-II subscales, a significantly higher score for “spiritual growth” and a significantly lower score for “physical activity” were seen in the agricultural group than in the non-agricultural group. In general, the old and female groups showed higher scores than the corresponding groups, regardless of job type.
It was determined that the major countermeasures to maintain a healthy lifestyle in agricultural workers should be associated with how to introduce daily activities that maintain and enhance “spiritual growth” and improve “physical activity”.
PMCID: PMC3047662  PMID: 21432222
Lifestyle; Health promotion; Quality of life; Health Promoting Lifestyle Profile-II; Agriculture
11.  Recovery of Cerebrovascular Reserves after Stenting for Symptomatic Carotid Artery Stenosis 
Interventional Neuroradiology  2010;16(4):420-428.
Although a decrease in cerebrovascular reserves (CVR) is known to enhance the risk of stroke, changes in this parameter after carotid artery stenting (CAS) have rarely been investigated. The present study is the first to compare CVR recoveries after applying CAS to patients with symptomatic carotid artery disease.
CAS was performed for 31 consecutive patients with symptomatic carotid artery disease. They underwent acetazolamide-challenged single photon emission computed tomography (SPECT) before and after CAS to obtain data on resting stage cerebral blood flow (CBFrest values) in various regions of interest (ROIs) defined by a three-dimensional stereotactic ROI template. CVR values on ipsilateral and contralateral hemispheric sides were then calculated based on the CBFrest data.
The 31 patients were dichotomized into unilateral (n=22) and bilateral (n=9) lesion groups, and no significant between-group differences were observed in CBFrest before and after CAS. In the unilateral group, there were no differences in CVR values before and after CAS. In the bilateral group, however, the CVR values significantly increased in nearly all the investigated ROIs on the contralateral side. Also, the hemispheric CVR values on both sides significantly increased after CAS in the bilateral group, while no such increase was observed in the unilateral group.
CAS in patients with symptomatic bilateral carotid artery disease has the potential utility for their haemodynamic improvement even on the contralateral hemispheric side.
PMCID: PMC3278007  PMID: 21162772
acetazolamide, carotid artery stenosis, cerebral blood flow, ischemic stroke, SPECT
12.  Calpain-mediated down-regulation of myelin-associated glycoprotein in lysophosphatidic acid-induced neuropathic pain 
Journal of neurochemistry  2010;113(4):1002-1011.
Lysophosphatidic acid receptor (LPA1) signaling initiates neuropathic pain through demyelination of the dorsal root (DR). Although LPA is found to cause down-regulation of myelin proteins underlying demyelination, the detailed mechanism remains to be determined. In the present study, we found that a single intrathecal (i.t.) injection of LPA evoked a dose- and time-dependent down-regulation of myelin-associated glycoprotein (MAG) in the DR through LPA1 receptor. A similar event was also observed in ex vivo DR cultures. Interestingly, LPA-induced down-regulation of MAG was significantly inhibited by calpain inhibitors (calpain inhibitor X, E-64 and E-64d) and LPA markedly induced calpain activation in the DR. The pre-treatment with calpain inhibitors attenuated LPA-induced neuropathic pain behaviors such as hyperalgesia and allodynia. Moreover, we found that sciatic nerve injury activates calpain activity in the DR in a LPA1 receptor-dependent manner. The E-64d treatments significantly blocked nerve injury-induced MAG down-regulation and neuropathic pain. However, there was no significant calpain activation in the DR by complete Freund’s adjuvant treatment, and E-64d failed to show anti-hyperalgesic effects in this inflammation model. The present study provides strong evidence that LPA-induced calpain activation plays a crucial role in the manifestation of neuropathic pain through MAG down-regulation in the DR.
PMCID: PMC2908211  PMID: 20420580
lysophosphatidic acid; neuropathic pain; myelin-associated glycoprotein; dorsal root; calpain; demyelination
13.  Description of environmental determinants of quality of life in children with intellectual disability in Japan using the Delphi technique 
The present study aimed to define the framework of an environment conducive to the well-being of children with intellectually disability (CID).
A questionnaire composed of 31 items was developed through literature review. Then a 2-round Delphi survey was conducted with 3 different panels: health professionals (HPs), parents of CID, and teachers. The participants were asked to rate each item, select and rank the 10 most important items, and suggest additional ones.
A total of 71 participants responded to the first round: 24 HPs, 22 parents, and 25 teachers. In the second round the overall response rate was 83%. At the end of the exercise, 12 items reached global consensus, i.e., in all groups. Only 5 items were ranked as most important by all groups: attitudes of family members at home; attitudes of HPs and teachers; support from family members at home; support at school (classmates and teachers); and government policies. Nevertheless, the panelists’ views diverged on the remaining items. Several additional elements were suggested.
The views of HPs, teachers, and parents are complementary for the improvement of quality of life (QOL) of CID. The present findings will be used as a basis for the development of an instrument to assess the living environment of CID.
PMCID: PMC2824106  PMID: 19806431
Quality of life; Intellectual disability; Child; Delphi technique; Environment
14.  Autotaxin and lysophosphatidic acid1 receptor-mediated demyelination of dorsal root fibers by sciatic nerve injury and intrathecal lysophosphatidylcholine 
Molecular Pain  2010;6:78.
Although neuropathic pain is frequently observed in demyelinating diseases such as Guillain-Barré syndrome and multiple sclerosis, the molecular basis for the relationship between demyelination and neuropathic pain behaviors is poorly understood. Previously, we found that lysophosphatidic acid receptor (LPA1) signaling initiates sciatic nerve injury-induced neuropathic pain and demyelination.
In the present study, we have demonstrated that sciatic nerve injury induces marked demyelination accompanied by myelin-associated glycoprotein (MAG) down-regulation and damage of Schwann cell partitioning of C-fiber-containing Remak bundles in the sciatic nerve and dorsal root, but not in the spinal nerve. Demyelination, MAG down-regulation and Remak bundle damage in the dorsal root were abolished in LPA1 receptor-deficient (Lpar1-/-) mice, but these alterations were not observed in sciatic nerve. However, LPA-induced demyelination in ex vivo experiments was observed in the sciatic nerve, spinal nerve and dorsal root, all which express LPA1 transcript and protein. Nerve injury-induced dorsal root demyelination was markedly attenuated in mice heterozygous for autotaxin (atx+/-), which converts lysophosphatidylcholine (LPC) to LPA. Although the addition of LPC to ex vivo cultures of dorsal root fibers in the presence of recombinant ATX caused potent demyelination, it had no significant effect in the absence of ATX. On the other hand, intrathecal injection of LPC caused potent dorsal root demyelination, which was markedly attenuated or abolished in atx+/- or Lpar1-/- mice.
These results suggest that LPA, which is converted from LPC by ATX, activates LPA1 receptors and induces dorsal root demyelination following nerve injury, which causes neuropathic pain.
PMCID: PMC2989310  PMID: 21062487
15.  Crystallization and preliminary X-ray crystallographic studies of the axin DIX domain 
The DIX domain of rat axin has been purified and crystallized. Crystals diffracted to 2.9 Å resolution using synchrotron radiation.
Axin is a negative regulator of the canonical Wnt signalling pathway that mediates the phosphorylation of β-catenin by glycogen synthase kinase 3β. The DIX domain of rat axin, which is important for its homooligomerization and interactions with other regulators in the Wnt pathway, was purified and crystallized by the sitting-drop vapour-diffusion technique using polyethylene glycol 6000 and lithium sulfate as crystallization agents. Crystals belong to space group P61 or P65, with unit-cell parameters a = b = 91.49, c = 84.92 Å. An X-ray diffraction data set has been collected to a nominal resolution of 2.9 Å.
PMCID: PMC2335085  PMID: 17554179
axin; DIX domain; Wnt signal; β-catenin
16.  Identification of prothymosin-α1, the necrosis–apoptosis switch molecule in cortical neuronal cultures 
The Journal of Cell Biology  2007;176(6):853-862.
We initially identified a nuclear protein, prothymosin-α1 (ProTα), as a key protein inhibiting necrosis by subjecting conditioned media from serum-free cultures of cortical neurons to a few chromatography steps. ProTα inhibited necrosis of cultured neurons by preventing rapid loss of cellular adenosine triphosphate levels by reversing the decreased membrane localization of glucose transporters but caused apoptosis through up-regulation of proapoptotic Bcl2-family proteins. The apoptosis caused by ProTα was further inhibited by growth factors, including brain-derived neurotrophic factor. The ProTα-induced cell death mode switch from necrosis to apoptosis was also reproduced in experimental ischemia-reperfusion culture experiments, although the apoptosis level was markedly reduced, possibly because of the presence of growth factors in the reperfused serum. Knock down of PKCβII expression prevented this cell death mode switch. Collectively, these results suggest that ProTα is an extracellular signal protein that acts as a cell death mode switch and could be a promising candidate for preventing brain strokes with the help of known apoptosis inhibitors.
PMCID: PMC2064059  PMID: 17353361
17.  Effects of Green Tea Fractions on Oxygen-Induced Retinal Neovascularization in the Neonatal Rat 
This study aimed to investigate the preventive effects of green tea fractions (GTFs) on rat model of oxygen-induced retinopathy (OIR). Neonatal Sprague-Dawley rats were exposed to daily cycles of 80% oxygen (20.5 h), ambient air (0.5 h), and progressive return to 80% oxygen (3 h) until postnatal day 12 (P12), then the rats were placed in ambient air until P18. The green tea was fractionated by DM-A50, DM-W, M-B, and M-W. The rats were treated once daily from P6 to P17 by gastric gavage of GTFs (0.05 or 0.01 g/ml) or distilled water (DW) at 50 µl/10 g body weight. On P18, the rats were sacrificed and the retinal samples were collected. The retinal neovascularization (NV) was scored and avascular areas (AVAs) were measured as a % of total retinal area (%AVAs) in ADPase stained retinas. The NV scores in 0.01 g/ml M-W were significantly lower than those in DW. The %AVAs in 0.05 g/ml DM-A50 and in 0.05 g/ml and 0.01 g/ml M-W were significantly lower than those in DW. There were less catechins, and less caffeine in M-W fraction compared with other GTFs, suggesting components of green tea except for catechins and caffeine might suppress the neovascularization in rat model of OIR.
PMCID: PMC2274993  PMID: 18392105
oxygen-induced retinopathy; green tea; neovascularization; retinopathy of prematurity
18.  Community-based analysis of the factorial structures of the recent increase in low birthweight infants 
This study was conducted to determine whether the regional factors were related to the increase in the percentage of low birthweight (LBW: <2,500g) infants in Kumamoto Pref., and to establish a tentative structure model for predicting low birthweight infants. Analyses for frequency of LBW infants between 1974 and 1997, and a multiple regression model and covariance structure model were conducted using data from the vital statistics between 1992 and 1997 and regional indicators concerned with LBW infants from official registered statistical data between 1992 and 1997. The 72 regional factors were clustered into four groups linked with agricultural areas such as Urban, Flat, Hilly and Mountainous areas. The recent increase in the incidence of LBW infants resulted from the increase in moderate-LBW (MLBW: 2,000-2,500 g) infants of full term-LBW infants. There was a steady annual increase in the Urban agricultural area LBW infants since 1992. The two structure analyses revealed that the Urban area had a marked effect on the increase in LBW infants, whereas, farm villages in Hilly or Moutainous areas had less effect on the increase in LBW infants. These findings suggest that the regional factors relating to the mothers’ life-style or regional environments play a key role in the etiology and prevention of LBW, and will be a useful in the analyses using official registered material.
PMCID: PMC2723582  PMID: 21432195
low birthweight infant; community-based factors; urban rural differences; structure model; agricultural area
20.  Recent trends in bagassosis in Japan. 
The only workers presently exposed to bagasse dust in Japan are the employees of sugar refineries and lacquerware factories. A follow up study of six former cases of bagassosis from among the retired employees of a paper board factory, closed since 1973, showed that none of the subjects still had bagassosis. Examinations of 70 employees of a sugar refinery for allergic reactions also showed no case of bagassosis. Seven cases with suspicious shadows of bagassosis on chest radiographs and four cases with positive serum precipitin to stored bagasse were, however, found among those 70 subjects. The results show the disappearance of a past episode of bagassosis and the possibility of a new occurrence of bagassosis among the employees of sugar refineries and lacquerware factories in the near future in Japan.
PMCID: PMC1039272  PMID: 1637710
21.  Allergy to laboratory animals: an epidemiological study. 
A large cross sectional survey was carried out using a self administered questionnaire to examine the prevalence of laboratory animal allergy (LAA) and the factors associated with its development. Out of 5641 workers who were exposed to animals at 137 laboratory animal facilities in Japan, 23.1% had one or more allergic symptoms related to laboratory animals. The commonest symptom as rhinitis. About 70% of LAA subjects developed symptoms during their first three years of exposure. Atopy (past and family history), the number of animal species handled, and the time spent in handling correlated significantly with the development of LAA as did some types of job. A close relation between nasal symptoms and exposure to rabbits and between skin symptoms and exposure to rats were found. LAA subjects developed symptoms most quickly to rabbits.
PMCID: PMC1039233  PMID: 1733454
22.  Effects of ML-236B (Compactin) on Sterol Synthesis and Low Density Lipoprotein Receptor Activities in Fibroblasts of Patients with Homozygous Familial Hypercholesterolemia 
Journal of Clinical Investigation  1981;67(5):1532-1540.
We studied biochemical genetics of low density lipoprotein (LDL) receptor mutations in fibroblasts from six homozygous and five heterozygous patients with familial hypercholesterolemia (FH). Three of six homozygotes are receptor-negative type and the other three homozygotes are receptor-defective type. In the cells from three receptor-negative homozygotes, the receptor binding, internalization, and degradation of 125I-LDL were 0.5±0.3 ng/mg protein (mean±SEM), 14±8 and 8±6 ng/mg protein per 6 h (four normal cells; 44±3, 386±32, and 1,335±214 ng/mg protein per 6 h), respectively. In the cells from three receptor-defective homozygotes, the receptor binding, internalization, and degradation of 125I-LDL were 6±2, 29±8, and 90±32 ng/mg protein per 6 h, respectively. In these six homozygotes, two pairs of siblings are included. Two siblings in the same family were classified as receptor-negative and two siblings in another family were classified as receptor-defective. The receptor-negative phenotypes and the receptor-defective phenotypes bred true in individual families. The cells from five heterozygotes showed ∼46% of the normal activities of receptor.
ML-236B, competitive inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase), completely inhibited the incorporation of [14C]acetate into digitonin-precipitable sterols in fibroblasts from normal subjects and heterozygous and homozygous patients with FH with the concentration of 0.5 μg/ml. However, at 0.05 μg/ml of ML-236B sterol synthesis in fibroblasts from homozygotes was not completely suppressed in contrast to normal and heterozygous cells. Moreover, after preincubation with 0.05 μg/ml of ML-236B for 24 h in medium containing lipoproteins, sterol synthesis in the cells from receptor-negative homozygote showed 75% of the initial activity compared with that of 25% without preincubation. In the cells from a normal subject and a heterozygote, sterol synthesis was inhibited even after preincubation. These results suggest that (a) the inhibitory effect of ML-236B is overcome in homozygote cells by their high intracellular levels of HMG-CoA reductase and (b) that a higher dose of ML-236B may be required to lower serum cholesterol levels in FH homozygotes than in heterozygotes.
PMCID: PMC370722  PMID: 7229037
23.  Follow-up studies on the distribution of haemoglobin levels in female farm workers. 
Haemoglobin concentrations in about 1000 women agricultural workers in Japan were measured every year, except in 1972, during the period 1967-77. Improvements were noted in the course of this investigation, and these were predominantly associated with the fact that those in the study community began to pay attention to the problem of low haemoglobin levels and to improve their diet, with an increase in daily food intake, particularly of animal protein and iron. In addition, a marked decrease in anaemia caused by hookworm also played an important role. In this paper, the change in haemoglobin concentration during the period of study are described and the aetiology is discussed.
PMCID: PMC1051972  PMID: 536678
24.  A bi-HAC vector system towards gene and cell therapy 
ACS synthetic biology  2014;3(2):83-90.
Genetic manipulations with mammalian cells often require introduction of two or more genes that have to be in trans-configuration. However, conventional gene delivery vectors have several limitations, including a limited cloning capacity and a risk of insertional mutagenesis. In this paper, we describe a novel gene expression system that consists of two differently marked HAC vectors containing unique gene loading sites. One HAC, 21HAC, is stably propagated during cell divisions, therefore, it is suitable for complementation of a gene deficiency. The other HAC, tet-O HAC, can be eliminated, providing a unique opportunity for transient gene expression, e.g. for cell reprogramming. Efficiency and accuracy of a novel bi-HAC vector system have been evaluated after loading of two different transgenes into these HACs. Based on analysis of transgenes expression and HACs stability in the proof of principle experiments, the combination of two HAC vectors may provide a powerful tool towards gene and cell therapy.
PMCID: PMC4137782  PMID: 25101815
human artificial chromosome vector; gene expression; gene and cell therapy
25.  Proposal for a New Noncontact Method for Measuring Tongue Moisture to Assist in Tongue Diagnosis and Development of the Tongue Image Analyzing System, Which Can Separately Record the Gloss Components of the Tongue 
BioMed Research International  2015;2015:249609.
Tongue diagnosis is a noninvasive diagnosis and is traditionally one of the most important tools for physicians who practice Kampo (traditional Japanese) medicine. However, it is a subjective process, and its results can depend on the experience of the physician performing it. Previous studies have reported how to measure and evaluate the shape and color of the tongue objectively. Therefore, this study focused on the glossy component in order to quantify tongue moisture in tongue diagnosis. We hypothesized that moisture appears as a gloss in captured images and measured the amount of water on the tongue surface in 13 subjects. The results showed a high correlation between the degree of gloss and the amount of water on the tongue surface and suggested that the moisture on the tongue can be estimated by the degree of gloss in a captured image. Because the moisture level on the tongue changes during the course of taking photos, it became clear that we had to wait at least 3 minutes between photos. Based on these results, we established the tongue image analyzing system (TIAS), which can consistently record the gloss and color of the tongue surface simultaneously.
PMCID: PMC4324740

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