Association between Insufficient Sleep and Dental Caries among Preschoolers in Japan: A Cross-Sectional Multicentre Study.

Recently, sleeping status has attracted attention for its relationship with oral health. In the present study, we have investigated the association between early childhood caries and sleeping status. A multicentre observational cross-sectional study was conducted among 332 preschoolers (aged 3-6 years) and their parents in Chitose, Japan. Dental caries and sleeping status were assessed in the children and the sleep quality and health literacy of the parents were also assessed. Univariate and multivariate regression analyses were used in order to investigate the effect of the sleeping status of the parents and their children on dental caries. Among the children, the prevalence of dental caries experience was 12.7%. The children without caries slept significantly longer and their parents had a better Pittsburgh Sleep Quality Index (PSQI) score than those with caries experience. The sleeping status and the numbers of caries in the children were significantly correlated. Health literacy was better in those without caries experience. Parents' PSQI was significantly positively correlated with the numbers of caries in the children (r = 0.19, p = 0.0004). The children's sleep durations, screen time, and parental smoking status were independently associated with early childhood caries. Poor sleeping status of children and their parents is related to dental caries among preschoolers.

PATZ1 knockdown enhances malignant phenotype in thyroid epithelial follicular cells and thyroid cancer cells.

This study was designed to examine the involvement of PATZ1 in carcinogenesis and dedifferentiation of thyroid cancer. Immunohistochemistry on clinical specimens indicated nuclear PATZ1 expression in all normal thyroid glands and adenomatous goiter, while nuclear PATZ1 expression decreased along with the dedifferentiation of thyroid cancer. Knockdown of nuclear PATZ1 by siRNA in an immortalized normal follicular epithelial cell line (Nthy-ori 3-1) altered cellular morphology and significantly increased cell proliferation, migration, and invasion. In addition, the expression of urokinase-type plasminogen activator (uPA), matrix metalloproteinase (MMP) 2, MMP9, and MMP11 was increased by PATZ1 knockdown in Nthy-ori 3-1 cells. When PATZ1 was silenced in differentiated thyroid cancer (DTC) cell lines (TPC-1 and FTC-133), proliferation, cellular motility, and expression of uPA and MMPs were significantly increased. Forced expression of exogenous PATZ1 decreased proliferation, cellular motility, and the expression of uPA and MMPs in ATC cell lines (ACT-1 and FRO). In thyroid cancer cell lines, PATZ1 functioned as a tumor suppressor regardless of p53 status. Moreover, the ratio of nuclear PATZ1 positive tumors was significantly decreased in ATC irrespective of p53 status. Our study demonstrates that PATZ1 knockdown enhances malignant phenotype both in thyroid follicular epithelial cells and thyroid cancer cells, suggesting that PATZ1 functions as a tumor suppressor in thyroid follicular epithelial cells and is involved in the dedifferentiation of thyroid cancer.

Coexpression of EpCAM, CD44 variant isoforms and claudin-7 in anaplastic thyroid carcinoma.

BACKGROUND: Anaplastic thyroid cancer is considered to be one of the most aggressive human malignancies, and the mean survival time after diagnosis is approximately six months, regardless of treatments. This study aimed to examine how EpCAM and its related molecules are involved in the characteristics of anaplastic thyroid carcinoma. METHODOLOGY/PRINCIPAL FINDINGS: Two differentiated thyroid cancer cell lines (TPC-1 and FTC-133), and two anaplastic thyroid cancer cell lines (FRO, ACT-1) were analyzed for expression of CD44 standard isoform (CD44s), CD44 variant isoforms, and EpCAM, and human aldehyde dehydrogenase-1 (ALDH1) enzymatic activity using flow cytometry. CD44s expression was higher in TPC-1 and FTC-133 than in the FRO and ACT-1, whereas ALDH1 activities were higher in FRO and ACT-1 than in TPC-1 and FTC-133. An inverse correlation between CD44s expression and ALDH1 activity was observed in all thyroid cancer cell lines. As for the expressions of CD44 variant isoforms, ACT-1 showed higher and FRO showed moderate CD44v6 expressions, whereas either TPC-1 or FTC-133 showed negative CD44v6 expression. EpCAM expressions in FRO and ACT-1 were higher than those in TPC-1 and FTC-133, and EpCAM expressions inversely correlated with those of CD44s. A positive correlation was observed between EpCAM expression and ALDH1 activity in thyroid cancer cell lines. In the RT-PCR analysis, the expression levels of EpCAM, caludin-7 and ALDH1 in FRO and ATC-1 cells were significantly higher than those in TPC-1 and FTC-133 cells. In clinical specimens of thyroid cancers, nuclear expression of EpCAM and high expression of CD44v6 were detected significantly more frequently in anaplastic carcinomas. CONCLUSIONS/SIGNIFICANCE: Our study suggests the possibility that EpCAM, together with CD44v6 and claudin-7 as well as ALDH1, may be involved in the development of the aggressive phenotype of anaplastic thyroid carcinoma. Our findings may suggest a novel therapeutic strategy for treatment of anaplastic thyroid carcinoma.

An efficient laboratory-scale preparative method for [1-(13)C]glycocholic acid.

A breath test using [1-(13)C]glycocholic acid as a substrate is a potential diagnostic method for small intestinal bacterial overgrowth syndrome. [1-(13)C]Glycocholic acid has been thus synthesized in an excellent yield from ethyl [1-(13)C]glycinate hydrochloride in a one-pot reaction. This method is suitable for the preparation of the labeled compound on a laboratory scale, which helps to perform extensive clinical studies of the breath test.

Gastric emptying in patients with autoimmune pancreatitis.

OBJECTIVES: Autoimmune pancreatitis (AIP) and its extrapancreatic lesions seem to be clinical manifestations of organs involved in IgG4-related systemic disease. To clarify whether the stomach is a target organ, gastric function was evaluated in patients with AIP. METHODS: In 6 patients with AIP, gastric emptying was assessed by Carbon 13 (¹³C) acetate breath test before and after steroid therapy. Based on 4-hour breath samples, the half ¹³CO2 excretion time (T(1/2)) and the time of maximal excretion (T(max)) were calculated as gastric emptying parameters. Data of 20 healthy volunteers were used as controls. The number of IgG4-positive plasma cells in gastrofiberscopic biopsy specimens was counted before and after steroid therapy. RESULTS: Both T(1/2) and T(max) in patients with AIP decreased significantly after steroid therapy (T(1/2): 1.89 ± 0.21 hours vs 1.69 ± 0.15 hours, P = 0.046; and T(max): 1.1 ± 0.2 hours vs 0.96 ± 0.2 hours, P = 0.027), and became similar to those of the controls (T(1/2): 1.69 ± 0.32 hours and T(max): 0.98 ± 0.2 hour). The number of IgG4-positive plasma cells infiltrating the gastric mucosa decreased after steroid therapy. CONCLUSIONS: Gastric emptying was impaired in patients with AIP and improved to the reference range after steroid therapy. The stomach may be a target organ of IgG4-related systemic disease.

The importance of characterization of FITC-labeled antibodies used in tissue cross-reactivity studies.

Fluorescein isothiocyanate (FITC)-labeled antibodies are widely used as primary antibodies in the tissue cross-reactivity (TCR) studies for the development of therapeutic antibodies. However, the effects of FITC-labeling on the characteristics of an antibody are poorly understood. The present study was performed to examine the effect of FITC-labeling on the binding affinity and immunohistochemical staining profile of an antibody, using several antibodies with different FITC-labeling indices. The results showed that the FITC-labeling index in antibody was negatively correlated with the binding affinity for its target antigen. Immunohistochemically, an antibody with a higher labeling index had a tendency to be more sensitive, but was also more likely to yield non-specific staining. Based on these findings, we recommend that a FITC-labeled antibody used as a primary antibody in a TCR study should be carefully selected from several differently labeled antibodies to minimize the decrease in the binding affinity and achieve the appropriate sensitivity and interpretation of the immunohistochemistry.

A brief outline of the history of the pancreatic anatomy.

In the middle of the 18th century, Kouan Kuriyama, a Japanese physician of the Choshu Domain, depicted the anatomy of the human pancreas in a report to his master, Toyo Yamawaki. This report is the first anatomical description of the pancreas in Japan. In the Mediterranean area, the pancreas was apparently first described about 2,000 years before his observation. Although there are quite a few reviews on the history of this complex organ, our brief essay offers a historical outline of the pancreas.

Selectively crosslinked hyaluronic acid hydrogels for sustained release formulation of erythropoietin.

A novel sustained release formulation of erythropoietin (EPO) was developed using hyaluronic acid (HA) hydrogels. For the preparation of HA hydrogels, adipic acid dihydrazide grafted HA (HA-ADH) was synthesized and analyzed with (1)H NMR. The degree of HA-ADH modification was about 69%. EPO was in situ encapsulated into HA-ADH hydrogels through a selective cross-linking reaction of bis(sulfosuccinimidyl) suberate (BS(3)) to hydrazide group (pK(a) = 3.0) of HA-ADH rather than to amine group (pK(a) > 9) of EPO. The denaturation of EPO during HA-ADH hydrogel synthesis was drastically reduced with decreasing pH from 7.4 to 4.8. The specific reactivity of BS(3) to hydrazide at pH = 4.8 might be due to its low pK(a) compared with that of amine. In vitro release of EPO in phosphate buffered saline at 37 degrees C showed that EPO was released rapidly for 2 days and then slowly up to 4 days from HA-ADH hydrogels. When the hydrogels were dried at 37 degrees C for a day, however, longer release of EPO up to 3 weeks could be demonstrated. According to in vivo release test of EPO from HA-ADH hydrogels in SD rats, elevated EPO concentration higher than 0.1 ng/mL could be maintained from 7 days up to 18 days depending on the preparation methods of HA-ADH hydrogels. There was no adverse effect during and after HA-ADH hydrogel implantation.

Digestion and absorption of patients with autoimmune pancreatitis.

BACKGROUND/AIMS: In patients with autoimmune pancreatitis, radiological, serological and pancreatic functional abnormalities usually improve with steroid therapy. However, no studies have specifically focused on digestion and absorption in such patients. This study aims to investigate digestion and absorption, in addition to pancreatic endocrine and exocrine function, and their responses to steroid therapy in patients with autoimmune pancreatitis. METHODOLOGY: Ideal body weight index, levels of serum albumin, total cholesterol and glucose, or glycosylated hemoglobin values were investigated in 22 patients with autoimmune pancreatitis. N-benzoyl-L-tyrosyl-paraminobenzoic acid excretion test was performed in 9 patients. In 10 patients treated with steroids, these factors were examined again about 2 months after medication. RESULTS: Levels of serum albumin decreased, while ideal body weight index and levels of serum albumin and cholesterol significantly increased after steroid therapy. Eleven (50%) patients showed evidence of diabetes mellitus, but this improved in 6 of 8 patients treated with steroids. Pancreatic exocrine function was reduced in 88% of patients. Impaired pancreatic exocrine function improved after steroid therapy in 3 of 7 patients treated. CONCLUSIONS: Mild malnutrition as well as impaired pancreatic endocrine and exocrine function were frequently detected in patients with autoimmune pancreatitis. Steroid therapy was occasionally effective for these conditions.

Carboxylester lipase gene polymorphism as a risk of alcohol-induced pancreatitis.

OBJECTIVES: Alcohol abuse causes pancreatic damage in humans. However, only 5% of alcoholic patients have a clinical manifestation of pancreatitis, and the genetic predisposition of alcohol-associated pancreatitis remains elusive. Nonoxidative metabolites of ethanol, fatty acid ethyl esters (FAEEs), might play an important role in pancreatic damage. Carboxylester lipase (CEL) has been known to catalyze FAEE synthesis from fatty acids and ethanol. METHODS: The variable number of tandem repeat (VNTR) polymorphism in the coding region of the CEL gene was studied in patients with alcoholic pancreatitis (n = 100), in alcoholics without pancreatitis (n = 52), in patients with nonalcoholic pancreatitis (n = 50), in hyperlipidemia patients (n = 96), and control subjects (n = 435). RESULTS: The frequency of the NN-type (wild-type) gene was significantly decreased in patients with alcoholic pancreatitis than in other groups. The frequency of subjects who had the L allele in patients with alcoholic pancreatitis was significantly higher than in other groups. The distribution of the CEL gene polymorphism was not different among the control subjects, alcoholics without pancreatitis, patients with nonalcoholic pancreatitis, and patients with hyperlipidemia. CONCLUSIONS: The CEL gene polymorphism, especially an increase in the frequency of the L allele, was found to be associated with alcohol-induced pancreatitis.

Impairment of host resistance to Listeria monocytogenes infection in liver of db/db and ob/ob mice.

Leptin is an adipocyte-derived hormone that regulates a number of physiological functions, including energy homeostasis and immune function. In immune responses, leptin plays a role in the induction of inflammation. We investigated a role of leptin in Listeria monocytogenes infection using leptin receptor-deficient db/db mice and leptin-deficient ob/ob mice. These mutant mice were highly susceptible to L. monocytogenes, and the elimination of bacteria from the liver was inhibited. After infection, the induction of monocyte chemoattractant protein-1 (MCP-1) and KC mRNA in the liver of db/db mice and the MCP-1 mRNA expression in the liver of ob/ob mice was decreased compared with their heterozygote littermates. Leptin replacement in ob/ob mice resulted in improvement of anti-listerial resistance and the MCP-1 mRNA expression. The elimination of L. monocytogenes was significantly enhanced, and the expression of MCP-1 and KC mRNA was completely reversed in db/db mice by insulin treatment. These results suggest that leptin is required for host resistance to L. monocytogenes infection and that hyperglycemia caused by leptin deficiency is involved in the inefficient elimination of bacteria from the liver. Moreover, defect of MCP-1 expression in the liver may be involved in the attenuated host resistance in these mutant mice.

The fermentation of different dietary fibers is associated with fecal clostridia levels in men.

Only a few reports have compared the fermentation of pectin and cellulose using the hydrogen-breath test, and no studies have examined the relation between the hydrogen breathing pattern and colonic microflora. Using breath-hydrogen measurements, we investigated whether different dietary fibers (DFs) were fermented differently and whether there were individual differences after ingestion of the same DF; we also examined the relation between individual fecal microflora and the fermentation of DF. Results of hydrogen tests in 14 men were compared after they had ingested 20 g of pectin, 20 g of cellulose, or 6 g of lactulose (a DF-like substance). We examined the relation between the breath hydrogen results and the subjects' fecal microflora. We defined significant fermentation (i.e., positive cases) as a continuous rise in hydrogen in the expiratory air of >19 ppm. The subjects were divided into 3 groups according to their hydrogen breath test pattern, i.e., positive for lactulose and pectin (Group LP, n = 4); positive for lactulose alone (Group L, n = 7); and negative for pectin, cellulose, and lactulose (Group N, n = 3). Individual differences were noted in subjects from Group LP and Group L. The detection frequency of lecithinase-negative clostridia was higher in Group LP than in the other groups (P < 0.05), and the detection frequency and the number of lecithinase-positive clostridia were higher in Groups LP and L than in Group N (P < 0.05). These findings suggest that the Clostridium species are associated with hydrogen production. The hydrogen breath test results of DFs depend on both the type of DF and the individual colonic microflora. The amount and constitution of colonic microflora might be predicted by the hydrogen-breath test using different DFs.

Chronic pancreatitis in the elderly in Japan.

BACKGROUND/AIM: Although the elderly comprise an increasingly large segment of the population, little has been written about chronic pancreatitis in this age group in Japan. In this study, we analyzed the clinical features of elderly Japanese patients with chronic pancreatitis and compared them with those of late-onset chronic pancreatitis patients in Western countries. METHODS: Subjects were 182 patients (162 males and 20 females) with chronic pancreatitis. They were divided into two groups: early-onset group (onset <65 years of age, n = 119) and late-onset group (onset > or =65 years of age, n = 63). Clinical findings and follow-up data were examined for each group. RESULTS: Alcohol abuse was the most common etiological factor in early-onset pancreatitis patients. In the late-onset group, the frequencies of idiopathic and autoimmune pancreatitis increased. Furthermore, the age at onset of autoimmune pancreatitis was >60 years in 94% of the cases. The late-onset group was more likely to have painless disease, and calcification of the pancreas and steatorrhea were rare in this group. Major causes of death were malignancy and malnutrition in each group. CONCLUSIONS: Early- and late-onset chronic pancreatitis showed different clinical features. Clinicians should consider autoimmune pancreatitis in the differential diagnosis in elderly patients with obstructive jaundice.

Pancreatic endocrine and exocrine function and salivary gland function in autoimmune pancreatitis before and after steroid therapy.

INTRODUCTION: Autoimmune pancreatitis (AIP) is a distinct clinical entity in which an autoimmune mechanism may be involved in pathogenesis. AIM: To investigate salivary gland function in addition to pancreatic endocrine and exocrine function in patients with AIP, and to determine changes occurring after steroid therapy. METHODOLOGY: Fasting serum glucose levels, oral glucose tolerance tests or glycosylated hemoglobin values were examined in 19 patients with AIP. N-benzoyl-L-tyrosyl-p-aminobenzoic acid excretion test, sialochemistry and parotid gland scintigraphy were performed in 8 patients. RESULTS: Eight patients had evidence of DM. Steroid therapy subsequently improved insulin secretion and glycemic control in 3 of 5 patients treated. Pancreatic exocrine function was reduced in 88% of patients. Impaired pancreatic exocrine function improved after steroid therapy in 3 of 6 patients treated. The 3 patients also showed treatment-related improvement in endocrine function. Concentration of beta2-microglobulin in saliva was significantly raised in patients with AIP compared with controls (P < 0.05). Ratio of cumulative peak count to injected radionuclide and washout ratio in parotid scintigraphy in patients with AIP was significantly lower than those of controls (P < 0.01). Salivary gland dysfunction improved after steroid therapy in all 5 patients treated. CONCLUSIONS: Pancreatic endocrine and exocrine and salivary gland function were frequently impaired in patients with AIP, and steroid therapy was occasionally effective for these dysfunctions.