Polymorphisms in the promoter regions of the CXCL1 and CXCL2 genes contribute to increased risk of alopecia areata in the Korean population.

Alopecia areata (AA) is a common disease, which causes hair loss in humans. AA has a genetically complex inheritance. This study investigated the possible correlations between single nucleotide polymorphisms (SNPs) in the promoter regions of the chemokine (C-X-C motif) ligand 1 (melanoma growth stimulating activity, alpha) (CXCL1) and chemokine (C-X-C motif) ligand 2 (CXCL2) genes and the development of AA in the Korean population. Two hundred and thirty-five AA patients and 240 control subjects were recruited. The specific SNPs occurring in the promoter regions of the CXCL1 and CXCL2 genes (rs3117604, -429C/T and rs3806792, -264T/C, respectively) were genotyped. All data obtained was evaluated using the SNPStats, SPSS 18.0, and the Haploview v.4.2 software platforms. The Odd's ratios (OR), 95% confidence intervals (CI), and P values were calculated using multiple logistic regression models. Analyses of the genetic sequences obtained revealed a significant correlation between the two SNPs and the development of AA (rs3117604, P = 0.0009 in co-dominant model 1, P = 0.01 in co-dominant model 2, P = 0.004 in the dominant model, P = 0.005 in the log-additive model, P = 0.012 in allele distribution; rs3806792, P = 0.036 in co-dominant model 2, P = 0.0046 in the log-additive model). The TT and CC haplotypes were also observed to show a significant association with increased risk of AA (TT haplotype, P = 0.0018; CC haplotype, P = 0.0349). Our data suggests that the CXCL1 and CXCL2 genes may be associated with AA susceptibility.

Assessment of masticatory function in patients with non-sagittal occlusal discrepancies.

Non-sagittal occlusal discrepancies such as posterior cross-bite and anterior openbite are common types of malocclusion, but studies on masticatory function related to those malocclusions have been scarce. The aim of this study was to quantify the masticatory performance in patients with non-sagittal discrepancies compared to those with normal occlusion, using both objective and subjective measures. Maximum bite force and contact area using Dental Prescale(®) system as a static objective assessment, Mixing Ability Index (MAI) as a dynamic objective evaluation and food intake ability (FIA) as a subjective assessment were analysed from 21 people in normal occlusion (Group N) and 64 patients with posterior cross-bite (Group C), anterior openbite (Group O) or both (Group B). The differences of the maximum bite force, the contact area, the MAI and the FIA were compared, and their correlations were figured out. The non-sagittal malocclusion groups showed lower values in the maximum bite force, the contact area, the MAI and the FIA compared to those in the normal group (P < 0·0001). Compared to Group N, Groups C, O and B showed 61·5%, 42·1% and 40·1% of the maximum bite force, and 84%, 84% and 76% of hard food FIA, respectively. However, there were no significant differences among Groups C, O and B. The MAI showed higher correlation with the FIA (r = 0·38, P < 0·01), than with the maximum bite force and the contact area (both r = 0·24, P < 0·5). These results revealed that masticatory function in patients with non-sagittal discrepancies is significantly reduced both objectively and subjectively.

Local application of periodontal ligament stromal cells promotes soft tissue regeneration.

OBJECTIVES: To test the potential stimulatory effect of local application of periodontal ligament (PDL) stromal cells on soft tissue regeneration. MATERIALS AND METHODS: Fluorescently labeled PDL cells outgrown from extracted human premolars or phosphate-buffered saline were locally injected to the cutaneous wounds created on mice. Soft tissue regeneration was evaluated for 14 days using photographs and histomorphometry. PDL cell engraftment was tracked with confocal microscopy. To detect the paracrine effect of the PDL cells on soft tissue regeneration, PDL cell-conditioned medium (CM) was evaluated for the concentration of secretory factors, transforming growth factor-beta 1 (TGFß1). The effect of PDL CM on the proliferation and migration of dermal fibroblast and keratinocyte was tested using MTT assay and migration assay. RESULTS: The application of PDL cells significantly promoted soft tissue regeneration compared with the application of PBS. Self-replicating PDL cells were engrafted into the hair follicles of the host tissue. Dermal fibroblast proliferation and keratinocyte migration were significantly enhanced by the treatment with PDL CM. Physiologically significant amount of TGFß1 was secreted from PDL cells into the CM. CONCLUSION: Local injection of PDL cells promoted soft tissue regeneration in part by the enhancement of fibroblast proliferation and keratinocyte migration through a paracrine mechanism.

Interleukin-13/-4-induced oxidative stress contributes to death of hippocampal neurons in aß1-42-treated hippocampus in vivo.

AIMS: The present study examined whether Aß(1-42) can induce endogenous expression of interleukin-13 (IL-13) or (IL-4) within activated microglia in the rat hippocampus in vivo. We further investigated whether these cytokines mediate ROS/RNS generation through activation of NADPH oxidase and/or inducible nitric oxide synthase (iNOS), and thus contribute to the degeneration of hippocampal neurons in vivo. RESULTS: Here, we show that IL-13 and IL-4, endogenously expressed in Aß(1-42)-activated microglia in hippocampus in vivo, contribute to degeneration of hippocampal neurons in vivo. Neutralization of IL-13 and IL-4 protected hippocampal neurons in vivo against neurotoxicity by inhibiting activation of microglial NADPH oxidase and iNOS, resulting in attenuation of ROS generation and oxidative damage of protein, lipid and DNA. INNOVATION: To our knowledge, this is the first study to demonstrate the possible involvement of endogenously expressed IL-13 and/or IL-4 in activated microglia after Aß(1-42) injection in the degeneration of hippocampal neurons in vivo. The current findings suggest that the deleterious effects of microglia-derived endogenous IL-13 and/or IL-4 are involved in oxidative stress-mediated neurodegenerative diseases, such as AD. CONCLUSION: We carefully hypothesize that IL-13 and IL-4, well-known as anti-inflammatory cytokines might serve as neurotoxic mediators by enhancing microglia-derived oxidative stress in Aß(1-42)-treated hippocampus in vivo.

Preparation of doxorubicin-containing chitosan microspheres for transcatheter arterial chemoembolization of hepatocellular carcinoma.

A new form of doxorubicin hydrochloride (DRH)-containing chitosan microspheres (CMs) was prepared by employing an expanding-loading-shrinking (E-L-S) process. One hundred mg of pre-formed CMs were soaked in absolute ethanol and then placed in reduced pressure (the expanding process). Ten mg of DRH (2 mg ml(-1)) were added into the expanded CMs (the loading process). Next the microspheres were freeze-dried (the shrinking process). As a result of this E-L-S process, 10% (w/w) DRH-containing CMs (DRH-CM) were made. During 7 days, 22.6% of the DRH was observed to be released on the in vitro drug release study. In addition, these new DRH-CMs could be used for transcatheter arterial chemoembolization (TACE) procedure in VX2 hepatic tumour models of rabbit and the anti-tumour effects of DRH-CMs were investigated. On the post-CT scan 7 days after the TACE, total infarctions of the VX2 tumour were observed in 5 rabbits among the 6 total rabbits.

Topiramate stimulates glucose transport through AMP-activated protein kinase-mediated pathway in L6 skeletal muscle cells.

The use of topiramate (TPM) in the treatment of binge-eating disorder, bulimia nervosa, and antipsychotic-induced weight gain has recently increased, however, the exact molecular basis for its effects on body weight reduction and improved glucose homeostasis, is yet to be elucidated. Here we investigated the effect and signaling pathway of TPM on glucose uptake in L6 rat skeletal muscle cells, which account for >70% of glucose disposal in the body. Intriguingly, we found that TPM (10 microM) stimulated the rate of glucose uptake up to twofold increase. And TPM-stimulated glucose transport was inhibited with the overexpression of dominant-negative form of AMP-activated protein kinase (AMPK), an important mediator in glucose transport, implicating that AMPK-mediated pathway is involved. The TPM-stimulated glucose transport was blocked by SB203580, a specific inhibitor of AMPK downstream mediator, p38 mitogen-activated protein kinase (MAPK) protein. LY294002, an inhibitor of phosphatidylinositol (PI) 3-kinase, which is another crucial mediator in independent glucose transport pathway, did not inhibit TPM-stimulated glucose transport. We also found that TPM increased the phosphorylation level of AMPK and p38 MAPK, whereas no effect on the activity of PI 3-kinase of TPM, when assessed by PI 3-kinase assay, was observed. These results together suggest that TPM stimulates glucose transport, not via PI 3-kinase mediated, but via AMPK-mediated pathway in skeletal muscle cells, thereby contributing to the body weight regulation and glucose homeostasis.

Expression of an artificial polypeptide with a repeated tripeptide glutamyl-tryptophanyl-lysine in Saccharomyces cerevisiae.

AIMS: Artificial genes, which encode 48 or 64 repeats of a tripeptide, glutamyl-tryptophanyl-lysine have been cloned to the yeast expression vector pAM82 containing the PHO5 promoter and expressed in Saccharomyces cerevisiae AH22. METHODS AND RESULTS: When the yeast cells harbouring recombinant plasmids pALTG6-2 and pALTG4-4 were derepressed in Burkholder minimal medium (Toh-e, A., Ueda, Y., Kakimoto, S.I. and Oshima, Y. (1973) Journal of Bacteriology113, 727-738) containing low phosphate (0.03 g l-1 KH2PO4 and 1.5 g l-1 KCl), the expression was the highest after 24 h induction and the artificial polypeptides were synthesized to about 10% (pALTG6-2) and 14% (pALTG4-4) of the total cell protein. CONCLUSIONS: The artificial polypeptides produced in yeast were made to react with the rabbit antiserum against the polypeptide purified from Escherichia coli and found only in the pellet fraction of cell lysates, indicating the formation of inclusion body. Artificial polypeptide consisting of Glu-Trp-Lys may be useful as partial supplement in food and feeds. SIGNIFICANCE AND IMPACT OF THE STUDY: The production of single cell enriched with homopolymers of an essential amino acid in yeast might be an important tool of supplementing cereal diets and feed grain rations and could be used as means for improvement of the amino acid profile of single cell protein and production of pharmaceutical peptides.

Evaluation of design parameters of osseointegrated dental implants using finite element analysis.

Finite element analyses were performed for various shapes of dental implant to study effects on stress distribution generated in the surrounding jaw bone and to determine an optimal thread shape for even stress distribution. It was found that the square thread shape filleted with a small radius was more effective on stress distribution than other dental implants used in the analyses. Additional analyses were performed on the implant with the thread shape obtained from previous analyses for varying other design parameters, such as the width of thread end and height of thread for various load directions, to determine the optimal dimensions of the implant. Stress distribution was more effective in the case when the width of thread end and the height of thread were 0.5p and 0.46p, respectively, where p is the screw pitch. Then, using the optimal implant thread dimensions determined previously, stress analyses were performed with various screw pitches and implant lengths, to investigate effects on stress distribution and to find the way to reduce the maximum effective stress generated in the jaw bone. Results show that the maximum effective stress decreased not only as screw pitch decreased gradually but also as implant length increased.

Idiopathic localized dilatation of the ileum: CT findings.

We report computed tomographic and pathologic findings of an adult case of idiopathic localized dilatation of the ileum presenting as hematochezia and bowel perforation. If a cyst-like structure that has narrow communications with proximal and distal bowel loops and a layered enhancement pattern similar to those of adjacent bowels on the computed tomogram of a patient with gastrointestinal bleeding, idiopathic localized dilatation of the ileum should be suspected.

Proton magnetic resonance chemical shift imaging (1H CSI)-directed stereotactic biopsy.

INTRODUCTION: To add metabolic information during stereotactic biopsy target selection, the authors adopted proton chemical shift imaging (1H CSI)-directed stereotactic biopsy. Currently, proton single voxel spectroscopy (SVS) technique has been reported in stereotactic biopsy. We performed 1H CSI in combination with a stereotactic headframe and selected targets according to local metabolic information, and evaluated the pathological results. PATIENTS AND METHOD: The 1H CSI-directed stereotactic biopsy was performed in four patients. 1H CSI and conventional Gd-enhancement stereotactic MRI were performed simultaneously after the fitting of a stereotactic frame. After reconstructing the metabolic maps of N-acetylaspartate (NAA)/phosphocreatine (Cr), phosphocholine (Cho)/Cr, and Lactate/Cr ratios, focal areas of increased Cho/Cr ratio and Lac/Cr ratios were selected as target sites in the stereotactic MR images. RESULTS: 1H CSI is possible with the stereotactic headframe in place. No difficulty was experienced performing 1H CSI or making a diagnosis. Pathological samples taken from areas of increased Cho/Cr ratios and decreased NAA/Cr ratios provided information upon increased cellularity, mitoses and cellular atypism, and facilitated diagnosis. Pathological samples taken from areas of increased Lac/Cr ratio showed predominant feature of necrosis. CONCLUSION: 1H CSI was feasible with the stereotactic headframe in place. The final pathological results obtained were concordant with the local metabolic information from 1H CSI. We believe that 1H CSI-directed stereotatic biopsy has the potential to significantly improve the accuracy of stereotactic biopsy targeting.

Laryngeal mask insertion during target-controlled infusion of propofol.

STUDY OBJECTIVE: To compare the Laryngeal Mask Airway (LMA; The Laryngeal Mask Airway Co., Ltd., Nicosia, Cyprus) insertion conditions produced by 6 and 8 microg/mL of target plasma concentrations (Cpt) during the induction of anesthesia with target-controlled infusion (TCI) of propofol. DESIGN: Randomized, prospective, single-blind, clinical study. SETTING: University hospital. PATIENTS: 44 ASA physical status I and II patients, 16 to 54 years of age, weighing between 45 and 100 kg, undergoing minor surgery in which the use of LMA was indicated. INTERVENTIONS: Patients were randomly divided into two groups (1 and 2) of 22 to compare the effects of different propofol concentrations. Three minutes after intravenous (IV) injection of midazolam 0.04 mg/kg, group 1 and 2 received TCI of propofol with 6 and 8 microg/mL of Cpt, respectively. LMA was inserted when the effect-site concentration (EC) reached 2.5 microg/mL, which was displayed on the infusion pump. MEASUREMENTS: The LMA insertion conditions (mouth opening, gagging, coughing, head or limb movement, laryngospasm, overall ease of insertion) were assessed, and hemodynamic responses were evaluated until 3 minutes after LMA insertion. Total dose of propofol, EC, and elapsed time since the start of TCI were recorded at five times: at the loss of consciousness and eyelash reflex, at 2.5 microg/mL of EC, and immediately, 1 minute, and 3 minutes after the insertion of LMA. MAIN RESULTS: There was no significant difference between the two groups in insertion conditions, despite the significantly larger total dose and shorter elapsed time (2.6 +/- 0.08 mg/kg and 109 +/- 5.0 s) in Group 2 than those (2.1 +/- 0.02 mg/kg and 140 +/- 4.1 s) in Group 1 at 2.5 microg/mL of EC (p < 0.05). Systolic and diastolic blood pressure decreased and heart rate increased significantly throughout the study period in both groups (p < 0.05). But there was a significant decrease in arterial pressure in Group 2 compared with Group 1 1 and 3 minutes after the insertion (p < 0.05). CONCLUSIONS: Induction with 8 microg/mL of Cpt, compared with 6 microg/mL, allowed earlier LMA insertion but, could not improve the conditions for LMA insertion and required more careful attention to the decrease in blood pressure after LMA insertion.

GT1b ganglioside induces death of dopaminergic neurons in rat mesencephalic cultures.

We examined neurotoxicity of GT1b against dopaminergic neurons in vitro. Cultures of mesencephalic cells deprived of serum underwent the loss of 19% of tyrosine hydroxylase immunopositive (TH-ip) neurons. In cultures deprived of serum, treatment with 10-30 microg/ml GT1b attenuated the number of TH-ip neurons by 26-69%, respectively, compared to non-treated cultures. Intriguingly, cultures deprived of serum were more vulnerable to GT1b-induced neurotoxicity. Application of 60 microg/ml GT1b to cultures grown in serum containing media resulted in the loss of 26% of TH-ip neurons, similar to that (28%) observed in serum-deprived cultures treated with 10 microg/ml GT1b. Moreover, in our cultures, absence of nitric oxide (NO) production after GT1b treatment was obvious. The present results strongly suggest direct neurotoxic actions of GT1b against dopaminergic neurons regardless of NO.

Effects of folate supplementation on two provisional molecular markers of colon cancer: a prospective, randomized trial.

OBJECTIVES: Dietary folate intake is inversely associated with the risk of colorectal cancer. This study investigated the effect of folate supplementation on genomic DNA methylation and DNA strand breaks in exons 5-8 of the p53 gene of the colonic mucosa, two provisional biomarkers of colon cancer. METHODS: Twenty subjects with adenomas were randomized to receive either folate (5 mg/day) or placebo for 1 yr after polypectomy. At baseline, 6 months and 1 yr, systemic and colonic measures of folate status were determined, as were the biomarkers mentioned earlier. RESULTS: Folate supplementation increased serum, red blood cell and colonic mucosal folate concentrations (p < 0.02). Folate supplementation also increased the extent of genomic DNA methylation at 6 months and 1 yr (p = 0.001), whereas placebo administration was associated with an increase in the extent of genomic DNA methylation only at 1 yr. Similarly, folate supplementation decreased the extent of p53 strand breaks in exons 5-8 at 6 months and 1 yr (p < 0.02), whereas placebo administration was associated with a decrease in the extent of p53 strand breaks only at 1 yr. CONCLUSIONS: Both of these provisional biomarkers of colon cancer underwent accelerated improvement at 6 months with folate supplementation. However, these markers also improved with placebo at 1 yr. Therefore, potential confounding factors that seem to modulate these biomarkers need to be identified and corrected in order for these markers to serve as suitable surrogate endpoints in folate chemoprevention trials.

Older men and women efficiently absorb vitamin B-12 from milk and fortified bread.

Nothing is directly known about the bioavailability of vitamin B-12 from dairy products or fortified grain products. We directly studied vitamin B-12 absorption from water, milk and fortified bread in adult subjects using (58)Co-labeled vitamin B-12 and a whole body gamma-ray counter/spectrophotometer. Sixteen healthy men and women over the age of 60 y with normal serum levels of vitamin B-12 and normal basal gastric acid secretion were studied. (58)Co vitamin B-12 (0.25 microg) was administered in water, milk or fortified bread to each subject along with 185 kBq (5.0 microCi) (51)Cr as a stool marker. Whole body counting was performed 30 min after ingestion of the radioactive dose and at 7 and 14 d after dosing. Mean absorptions from water, milk and fortified bread were 55, 65 and 55%, respectively, and did not differ. The high body retention of the extrinsic vitamin B-12 label from milk and bread may warrant a greater use of such fortified products in the elderly to ensure vitamin B-12 adequacy.

Accurate measures of left ventricular ejection fraction using electron beam tomography: a comparison with radionuclide angiography, and cine angiography.

BACKGROUND: Quantitative determination of ejection fraction is predicated on precise measurement of end-diastolic and end-systolic volumes of the left ventricle. Contrast enhanced electron beam tomography (EBT), with excellent temporal and spatial resolution, has the potential for highly accurate measures of ejection fraction. METHODS: EBT protocol used a short axis scan of the left ventricle (8-12 levels, apex to base) during infusion of iodinated contrast. To assess the accuracy of the measured left ventricular ejection fraction (LVEF), we compared EBT with first-pass radionuclide angiography (RNA) and cine angiography (CINE). RESULTS: A total of 41 patients (26 men and 15 women) underwent all three tests within 1 week. Resting ejection fraction using each modality was assessed in a linear regression model to assess inter-test correlation with the other two modalities. Correlation between CINE and EBT was high (r = 0.90, intercept 4.67, p < 0.001). Similarly, correlation of CINE and RNA (r = 0.87, intercept -5.48, p < 0.001) and between EBT and RNA (r = 0.87, intercept -4.6, p < 0.001) were high. In a subset of those patients with LVEF < or = 40%, correlation was consistently high between EBT and CINE. However, correlations were poor for the comparisons between RNA and CINE (r = 0.40), and between the RNA and EBT (r = 0.47). The mean differences of measured ejection fractions between each of the imaging modality were small. However, there was only modest agreement between each of the comparisons as measured using 95% confidence interval (CI) on Bland-Altman plots. CONCLUSION: These data indicate that the LVEF results are comparable among EBT, RNA, and CINE and can be used interchangeably to assess ventricular function for LVEF > 40%. For LVEF < or = 40%, we demonstrated some disparate results between cine angiography and RNA and between EBT and RNA, indicating that CINE or EBT may provide more accurate assessment.

Cephalometric characteristics of Korean Class III surgical patients and their relationship to plans for surgical treatment.

In comparison to gender-matched normal Koreans, Korean patients selected for surgical correction of skeletal Class III problems have, on average, a shorter anterior and posterior cranial base, a shorter maxilla, a longer mandible, increased lower facial height, and a retrusive upper lip. In both males and females, about 40% of a group of Class III patients scheduled for surgery had a maxilla within one standard deviation of the normal position and a prognathic mandible, as compared with a group of normal Korean adults. Almost as many males (37%) in the surgical group had both a retrognathic maxilla and a prognathic mandible, while 18% had a retrognathic maxilla and normal mandible. In females, 25% had only a retrognathic maxilla and 25% had both jaws outside the normal range. The percentage of the Korean patients whose Class III relationship was primarily a result of mandibular prognathism (48%) is more than twice as high as the corresponding number for American Class III surgical patients (19%), somewhat higher than in Chinese patients (39%), and similar to the percentage of Japanese (50%). Maxillary surgery, alone or in conjunction with mandibular setback, is currently used in the treatment of most Class III patients. Both the esthetic consideration of widening the already broad Asian nose and the relative proportions of maxillary versus mandibular abnormalities suggest that mandibular setback alone can be considered for a higher number of Asian than Caucasian Class III patients.

Chronic alcohol consumption induces genomic but not p53-specific DNA hypomethylation in rat colon.

Alcohol consumption has been implicated as an etiologic agent in colorectal carcinogenesis, but the mechanism by which alcohol enhances the development of colorectal cancer is not yet known. Recent reports indicate that alcohol consumption can diminish cellular S-adenosylmethionine levels, thus possibly altering normal patterns of DNA methylation, a phenomenon that is mediated by S-adenosylmethionine and whose abnormalities are observed in colonic neoplasia. This study investigated the effect of chronic alcohol consumption on genomic DNA methylation of rat colonic epithelium and methylation of the p53 tumor suppressor gene, abnormalities of which have been implicated in colonic carcinogenesis. Two groups of rats (n = 10/group) were pair-fed either an alcohol-containing or an isocaloric control Lieber-DeCarli diet for 4 wk. The extent of genomic DNA methylation was assessed by incubating the extracted DNA with [(3)H]S-adenosylmethionine and Sss1 methyltransferase. Gene-specific methylation was assessed by using semiquantitative polymerase chain reaction (PCR). Tritiated methyl uptake by colonic DNA (which is inversely correlated with genomic methylation) from alcohol-fed rats was 57% less than that in control DNA (P < 0.05). However, gene-specific DNA methylation, both in the p53 gene (exons 5-8) and in the beta-actin gene, a control gene, did not differ between the two groups. In conclusion, this study indicates that chronic alcohol consumption produces genomic DNA hypomethylation in the colonic mucosa. This may constitute a means by which carcinogenesis is enhanced, although further studies are required to establish causality.

Altered presynaptic gene expression in transgenic mice producing dopamine in the pineal gland.

Neurotransmitters are known to play an important role in the development of the nervous system. We recently generated transgenic mice that ectopically express tyrosine hydroxylase (TH) and thereby produce dopamine (DA) de novo in pinealocytes of the pineal gland (PG). The transgenic PG also exhibited a dramatic decrease in TH-immunoreactive (IR) fibers putatively arising from the superior cervical ganglion (SCG) (Cho et al. [1996] Proc Natl Acad Sci USA 93:2862-2866). In the current study, however, we found that there was no reduction in the number of fibers immunostained for neurofilament protein or PGP9.5, markers known to be heavily localized in fibers, despite the reduction of TH fiber density. Therefore, we investigated whether the decreased TH-IR fiber density is the consequence of reduced sympathetic innervation, or a decrease in TH expression within innervating fibers. Immunohistochemical analysis comparing control and transgenic PG demonstrated no apparent differences in numbers of NPY- and aromatic-L-amino acid decarboxylase (AADC)-IR fibers, indicating that TH expression is decreased in a normal number of innervating fibers. Furthermore, presynaptic neurons in the transgenic SCG showed abnormal and heterogeneous TH immunoreactivity and reduced TH and norepinephrine transporter (NET) mRNA levels. These results show that ectopic DA production in the PG lowers TH and NET gene expression in the SCG without altering sympathetic innervation to the PG and suggest that the alteration of target neurotransmitter phenotype may influence gene expression of phenotype-specific proteins in projecting neurons.