Novel presynaptic assay system revealed that metformin ameliorates exaggerated synaptic release and Munc18-1 accumulation in presynapses of neurons from Fragile X syndrome mouse model.

Fragile X syndrome (FXS) is a developmental disorder characterized by intellectual disability and autistic-like behaviors. These symptoms are supposed to result from dysregulated translation in pre- and postsynapses, resulting in aberrant synaptic plasticity. Although most drug development research on FXS has focused on aberrant postsynaptic functions by excess translation in postsynapses, the effect of drug candidates on FXS in presynaptic release is largely unclear. In this report, we developed a novel assay system using neuron ball culture with beads to induce presynapse formation, allowing for the analysis of presynaptic phenotypes, including presynaptic release. Metformin, which is shown to rescue core phenotypes in FXS mouse model by normalizing dysregulated translation, ameliorated the exaggerated presynaptic release of neurons of FXS model mouse using this assay system. Furthermore, metformin suppressed the excess accumulation of the active zone protein Munc18-1, which is supposed to be locally translated in presynapses. These results suggest that metformin rescues both postsynaptic and presynaptic phenotypes by inhibiting excess translation in FXS neurons.

[Derivation of Conversion Formula of Image Quality Figure (IQFinv) Values by Old and New Analysis Software for Mammographic Contrast-detail Phantom].

PURPOSE: This study deals with the conversion of the image quality figure (IQFinv) values with CDMAM Analyser ver. 1 (old analysis software) used to analyse the images of CDMAM 3.4 phantom into the IQFinv values with a new analysis software, that is, CDMAM Analyser (ver. 2 and ver. 2.3), which is a software improved to be closer to human visual evaluation. METHODS: The IQFinv values were calculated using four mammography units and three analysis software. The IQFinv values using the old and new software were compared. RESULTS: The IQFinv values had no difference according to mammography units and beam quality. The conversion formula of IQFinv from CDMAM Analyser ver. 1 to CDMAM Analyser ver. 2 was derived. Furthermore, the conversion formula of IQFinv from CDMAM Analyser ver. 1 to CDMAM Analysis ver. 2.3 was also derived. CONCLUSION: Using each conversion formula, the IQFinv values using the old software can be converted to those using the new software. There was a slight difference between the IQFinv values using the new software.

[Evaluation of the Capacity to Produce Histamine by Histamine-Producing Bacteria during Storage at 10℃].

Histamine is produced from histidine using histidine decarboxylase of histamine-producing bacteria. However, associated histamine food poisoning demands microbiological controls. Furthermore, studies reported that histamine production by histamine-producing bacteria is affected by temperature. Therefore, to prevent histamine food poisoning, it is desirable to store foods below 4℃. However, it is challenging to maintain the storage temperature of food substances in refrigerators constantly below 4℃. Thus, we investigated histamine production capacity using seven histamine-producing bacterial strains under storage at 10℃, a more reasonable cold storage condition. Subsequently, we examined the variation of histamine production in buffers, the correlation between bacterial density and histamine production quantities, and the growth rate in broths. Results showed that similar levels of histamine were produced in buffers even after 5 days of storage under certain conditions in which histamine-producing bacteria did not grow. Moreover, bacterial density was proportional to histamine production, and the coefficient of determination was more than 0.97, and the bacterial density required to produce 200 µg/mL of histamine during storage at 10℃ was calculated to be 4×107-4×108 CFU/mL. When the initial bacterial density was 102-103 CFU/mL, psychrophilic bacteria required 2 or 3 days and mesophilic bacteria required more than 4 days to grow above 107 CFU/mL. The above results suggest that understanding the capacity of histamine-producing bacteria to produce histamine and its growth rate in foods is important for the prevention of histamine food poisoning.

[Prevalence of Foodborne Pathogens in Retail Meat in Saitama, Japan].

We investigated the prevalence of Campylobacter jejuni, Campylobacter coli, Salmonella, enterohemorrhagic Escherichia coli, enterotoxigenic Escherichia coli, Yersinia enterocolitica, and Escherichia albertii in domestic chicken and pork sold at retail stores in Saitama Prefecture, Japan. Campylobacter was detected in 35.7% (60/168) of chicken samples and 7.3% (14/190) of pork samples. C. jejuni and C. coli were predominant in chicken and pork, respectively. Salmonella was found in 58.1% (100/172) of chicken samples and 19.9% (41/206) of pork samples. Moreover, Salmonella Schwarzengrund was the major serovar observed in chicken isolates, whereas S. Typhimurium monophasic variant was in pork isolates. Furthermore, ETEC was found in 0.6% (1/160) of chicken samples and 2.4% (5/206) of pork samples. Y. enterocolitica was absent from all (83/83) chicken samples but was present in 9.3% (18/193) of pork samples, with a prevalence in pork tongues as high as 21.0% (13/62 samples). However, EHEC and E. albertii were not detected in our study. Therefore, the results of this study indicate that chicken was highly contaminated with Campylobacter sp. and Salmonella, and pork was with Y. enterocolitica serotype O3, Campylobacter sp., Salmonella, and ETEC.

A Farnesyltransferase Inhibitor Restores Cognitive Deficits in Tsc2+/- Mice through Inhibition of Rheb1.

Tuberous sclerosis complex (TSC) is caused by mutations in Tsc1 or Tsc2, whose gene products inhibit the small G-protein Rheb1. Rheb1 activates mTORC1, which may cause refractory epilepsy, intellectual disability, and autism. The mTORC1 inhibitors have been used for TSC patients with intractable epilepsy. However, its effectiveness for cognitive symptoms remains unclear. We found a new signaling pathway for synapse formation through Rheb1 activation, but not mTORC1. Here, we show that treatment with the farnesyltransferase inhibitor lonafarnib increased unfarnesylated (inactive) Rheb1 levels and restored synaptic abnormalities in cultured Tsc2+/- neurons, whereas rapamycin did not enhance spine synapse formation. Lonafarnib treatment also restored the plasticity-related Arc (activity-regulated cytoskeleton-associated protein) expression in cultured Tsc2+/- neurons. Lonafarnib action was partly dependent on the Rheb1 reduction with syntenin. Oral administration of lonafarnib increased unfarnesylated protein levels without affecting mTORC1 and MAP (mitogen-activated protein (MAP)) kinase signaling, and restored dendritic spine morphology in the hippocampi of male Tsc2+/- mice. In addition, lonafarnib treatment ameliorated contextual memory impairments and restored memory-related Arc expression in male Tsc2+/- mice in vivo Heterozygous Rheb1 knockout in male Tsc2+/- mice reproduced the results observed with pharmacological treatment. These results suggest that the Rheb1 activation may be responsible for synaptic abnormalities and memory impairments in Tsc2+/- mice, and its inhibition by lonafarnib could provide insight into potential treatment options for TSC-associated neuropsychiatric disorders.SIGNIFICANCE STATEMENT Tuberous sclerosis complex (TSC) is an autosomal-dominant disease that causes neuropsychiatric symptoms, including intractable epilepsy, intellectual disability (ID) and autism. No pharmacological treatment for ID has been reported so far. To develop a pharmacological treatment for ID, we investigated the mechanism of TSC and found that Rheb1 activation is responsible for synaptic abnormalities in TSC neurons. To inhibit Rheb1 function, we used the farnesyltransferase inhibitor lonafarnib, because farnesylation of Rheb1 is required for its activation. Lonafarnib treatment increased inactive Rheb1 and recovered proper synapse formation and plasticity-related Arc (activity-regulated cytoskeleton-associated protein) expression in TSC neurons. Furthermore, in vivo lonafarnib treatment restored contextual memory and Arc induction in TSC mice. Together, Rheb1 inhibition by lonafarnib could provide insight into potential treatments for TSC-associated ID.

[A Method to Obtain a Sheet-like X-ray Phantom Made from Iodine Contrast Agent].

PURPOSE: A sheet-like X-ray phantom on which thin Iodine is uniformly coated was developed to facilitate the handling of iodined objects used in any X-ray imaging studies. METHODS: The most recommendable protocol as follows: (1) prepare undiluted 240 mg/ml Iohexol-based contrast agent and drop around 1.6 g on a horizontal surface. (2) infiltrate the agent into a membrane filter (47 mm in diameter) from the secondary side. (3) one minute later, the excess liquid components should be removed by a softy absorbent paper, and the infiltrated membrane filter should be left until substantially dried. RESULT: The dried membrane filter can be utilized as a sheet-like X-ray phantom on which the iodine of around 2.45 mg is almost uniformly distributed per 1 cm2. However, since the iodine density is slightly higher on the periphery part of the sheet, less than 80% area of the entire sheet is recommended to be used from a viewpoint of the spatial uniformity. CONCLUSION: In the practical experiments, the X-ray attenuation factor can be controlled by changing the stacking number of the sheet, and the spatial size and form can also be designed by cutting the sheet. This capability is expected to improve the efficiency of any X-ray experiments and quality control works that requires iodined materials.

Myosin VI Haploinsufficiency Reduced Hearing Ability in Mice.

In human, myosin VI (MYO6) haploinsufficiency causes postlingual progressive hearing loss. Because the usefulness of mouse models remains unclear, we produced novel Myo6 null (-/-) mutant mice and analyzed the hearing phenotypes of Myo6+/- (+/-) heterozygous mutants. We first recorded and compared the auditory brainstem responses and distortion product otoacoustic emissions in control Myo6+/+ (+/+) wild-type and +/- mice. These hearing phenotypes of +/- mice were mild; however, we confirmed that +/- mice developed progressive hearing loss. In particular, the hearing loss of female +/- mice progressed faster than that of male +/- mice. The stereocilia bundles of +/- mice exhibited progressive taper loss in cochlear inner hair cells (IHCs) and outer hair cells (OHCs). The loss of OHCs in +/- heterozygotes occurred at an earlier age than in +/+ mice. In particular, the OHCs at the basal area of the cochlea were decreased in +/- mice. IHC ribbon synapses from the area at the base of the cochlea were significantly reduced in +/- mice. Thus, our study indicated that MYO6 haploinsufficiency affected the detection of sounds in mice, and we suggest that +/- mice with Myo6 null alleles are useful animal models for gene therapy and drug treatment in patients with progressive hearing loss due to MYO6 haploinsufficiency.

[Construction and Analysis of a Digital Mammography Database: Analysis of Breast Compression Pressures at Four Facilities].

We analyzed the compression pressures in 2772 mammography images of 807 patients acquired by digital mammography equipment at four facilities. The analysis included the average compression pressure at all facilities, difference in compression pressure at each facility, differences between the pressures used by radiological technologists in the same facility, and difference attributed to the breast structure. We also analyzed the effects of the compression pressure on the breast thickness and mean glandular dose (MGD) at each facility. The median values of the compression pressure and breast thickness for the 2772 images at all facilities were 86.5 N and 43 mm, respectively. The compression pressures differed among the institutions. The maximum difference in the median pressures among the four facilities was 38.6 N, while the difference in the breast thickness was 6 mm. The radiological technologists working at the same facility used almost the same compression pressure. However, differences between the compression pressures used by different radiological technologists were observed. The compression pressure in a dense breast was smaller than that in a non-dense breast. The difference in the compression pressure affected the breast thickness and MGD. The results of this analysis could be utilized for an optimal imaging in future digital mammography.

Coronin-1 is phosphorylated at Thr-412 by protein kinase Cα in human phagocytic cells.

Coronin-1, a hematopoietic cell-specific actin-binding protein, is thought to be involved in the phagocytic process through its interaction with actin filaments. The dissociation of coronin-1 from phagosomes after its transient accumulation on the phagosome surface is associated with lysosomal fusion. We previously reported that 1) coronin-1 is phosphorylated by protein kinase C (PKC), 2) coronin-1 has two phosphorylation sites, Ser-2 and Thr-412, and 3) Thr-412 of coronin-1 is phosphorylated during phagocytosis. In this study, we examined which PKC isoform is responsible for the phosphorylation of coronin-1 at Thr-412 by using isotype-specific PKC inhibitors and small interfering RNAs (siRNAs). Thr-412 phosphorylation of coronin-1 was suppressed by Gö6976, an inhibitor of PKCα and PKCßI. This phosphorylation was attenuated by siRNA for PKCα, but not by siRNA for PKCß. Furthermore, Thr-412 of coronin-1 was phosphorylated by recombinant PKCα in vitro, but not by recombinant PKCß. We next examined the effects of Gö6976 on the intracellular distribution of coronin-1 in HL60 cells during phagocytosis. The confocal fluorescence microscopic observation showed that coronin-1 was not dissociated from phagosomes in Gö6976-treated cells. These results indicate that phosphorylation of coronin-1 at Thr-412 by PKCα regulates intracellular distribution during phagocytosis.

[Comparison between Variance and Edge Methods for Measuring Image Resolution Properties in Digital Mammography].

We studied the image resolution properties in digital mammography using the variance and edge response function methods. The associated measurements were made using a polymethylmethacrylate phantom of 0-40 mm thickness while maintaining the incident dose to the detector constant, as well as with and without using the phantom but by varying the level of exposure to the detector. The results obtained using the two methods were compared on the object plane of the mammography unit with compensation for the magnification and rejection of the scattered radiation. In this unit, the imaging process of resolution enhancement may be performed according to the phantom thickness, shape of test device, exposure level, and so on. The modulation transfer function values of the variance method were slightly higher than those of the edge method, approximately 6% at 2 cycles/mm and 8% at 4 cycles/mm without resolution enhancement processing. We concluded that the variance method is more suitable than the edge method for the constancy test or quality control because of its lower coefficient of variation.

Residual Analysis of Aflatoxins in Spice by HPLC Coupled with Solid-Phase Dispersive Extraction and Solid-Phase Fluorescence Derivatization Method.

BACKGROUND: Aflatoxins (AFs) are carcinogenic mycotoxins. A simple, quick, and accurate method for the micro-analysis of AFs in foodstuffs, especially spices, is needed. OBJECTIVE: A sophisticated pretreatment method that combines solid-phase dispersive extraction (SPDE) and solid-phase fluorescence derivatization using immunoaffinity (IA) gel as the solid phase was developed to analyze AFs in spices simply, quickly, and sensitively by liquid chromatography with fluorescence detection. METHOD: White and black pepper samples were extracted with a mixed solution of methanol/water (4:1) and then diluted with 7% aqueous solution of Triton-X. The solution was subjected to cleanup by SPDE using IA gel. Trifluoroacetic acid was added to the IA gel for on-site solid-phase fluorescence derivatization. RESULTS: Chromatograms containing well-separated peaks and few interference peaks from contaminants were obtained. The method detection limit of AFs in white and black pepper was 0.15-0.29 ng/g. Repeatability and intermediate precision were <10% and <15%, respectively, and accuracy was 61.7-87.8%. In addition, inter-laboratory precision was <29% and mean recovery was 61.5-76.7%. A favorable z-score of |Z| ≦ 1 was obtained in seven laboratories, although one laboratory gave 2 < |Z| < 3. CONCLUSIONS: The validity, reliability, practicality, and robustness of the developed method were verified. HIGHLIGHTS: By using SPDE and solid-phase fluorescence derivatization in combination for AF analysis, fluorescence derivatization during cleanup was realized, leading to simplification of the pretreatment operation.

A cross-cultural study of acceptability and food pairing for hot sauces.

This study was conducted to understand the acceptance levels of hot sauces among consumers from different culinary cultures. Two newly developed hot sauces [fermented red chili pepper with soybean-paste-based sauce(GS) and fermented red chili-pepper-based sauce(KS)] were compared with Tabasco sauce(TB) and Sriracha sauce(SR). Two separate cross-cultural home-use tests(HUTs) were conducted: pizza and cream soup were provided as food items in HUT 1, whereas grilled chicken wings and rice noodle soup were provided in HUT 2. Consumers residing in Denmark, South Korea, and US participated in each HUT (n≅100 per country). Acceptance levels and the reasons for (dis)liking particular hot sauces applied to food systems were assessed. The food items that paired well with different hot sauces when the sauces were applied freely to regular meals were also analyzed among the US and Korean subjects. When the hot-sauce samples were applied to pizza and cream soup, the preferred order of the samples exhibited a cross-cultural agreement (GS = KS > TB). In the case of grilled chicken and rice noodle soup, the acceptance rating was similar for the three types of hot sauces among Koreans, whereas the acceptance was higher for SR among the US subjects for both foodstuffs, while Danish subjects preferred GS and KS over SR. The US subjects did not like hot-sauce samples with sweet and weak spiciness, whereas the Korean and Danish subjects disliked the hot-sauce sample when it was too spicy and not sufficiently sweet. These findings indicate that the matching of particular sauces with specific food items is culture-dependent, and this needs to be considered when trying to export food products such as hot sauce to other countries.

Evidence of exposure to chemicals and heavy metals during pregnancy in Japanese women.

Purpose: Prenatal exposure to environmental chemicals is a growing concern, because such exposures have been shown to be associated with various diseases. The levels of chemicals and heavy metals in maternal blood, cord blood, maternal urine and amniotic fluid in Japanese pregnant women were investigated. Methods: A total of 145 women, including 14 fetal growth restriction cases, were included in the present study. The levels of phthalates (di[2-ethylhexyl]phthalate and mono[2-ethylhexyl]phthalate), perfluorinated compounds (perfluorooctane sulfonate, perfluorohexanoic acid, perfluorooctanoic acid, and perfluorononanoic acid), pesticides (dimethylphosphate, dimethylthiophosphate, diethylphosphate, diethylthiophosphate, 3-phenoxybenzoic acid, and octachlorodipropyl ether), bisphenol A, nicotine (nicotine, nornicotine, cotinine, norcotinine, and trans-3'-hydroxycotinine), polybrominated diphenyl ethers, and heavy metals were measured. The relationship between fetal growth and the levels of chemicals and heavy metals were investigated. Results: Phthalates, perfluorinated compounds, pesticides, polybrominated diphenyl ethers, and heavy metals were detected in high frequency, whereas nicotine and bisphenol A were almost negative. Phthalates, perfluorinated compounds, and several heavy metals were transferred to the fetus. High perfluorononanoic acid levels in the maternal blood and cord blood, and low perfluorooctanoic acid level in the cord blood were significantly and negatively associated with fetal growth. Conclusions: The present study showed that pregnant women in Japan and their fetuses are exposed to a variety of chemicals and heavy metals.

A novel splice site mutation of myosin VI in mice leads to stereociliary fusion caused by disruption of actin networks in the apical region of inner ear hair cells.

An unconventional myosin encoded by the myosin VI gene (MYO6) contributes to hearing loss in humans. Homozygous mutations of MYO6 result in nonsyndromic profound congenital hearing loss, DFNB37. Kumamoto shaker/waltzer (ksv) mice harbor spontaneous mutations, and homozygous mutants exhibit congenital defects in balance and hearing caused by fusion of the stereocilia. We identified a Myo6c.1381G>A mutation that was found to be a p.E461K mutation leading to alternative splicing errors in Myo6 mRNA in ksv mutants. An analysis of the mRNA and protein expression in animals harboring this mutation suggested that most of the abnormal alternatively spliced isoforms of MYO6 are degraded in ksv mice. In the hair cells of ksv/ksv homozygotes, the MYO6 protein levels were significantly decreased in the cytoplasm, including in the cuticular plates. MYO6 and stereociliary taper-specific proteins were mislocalized along the entire length of the stereocilia of ksv/ksv mice, thus suggesting that MYO6 attached to taper-specific proteins at the stereociliary base. Histological analysis of the cochlear hair cells showed that the stereociliary fusion in the ksv/ksv mutants, developed through fusion between stereociliary bundles, raised cuticular plate membranes in the cochlear hair cells and resulted in incorporation of the bundles into the sheaths of the cuticular plates. Interestingly, the expression of the stereociliary rootlet-specific TRIO and F-actin binding protein (TRIOBP) was altered in ksv/ksv mice. The abnormal expression of TRIOBP suggested that the rootlets in the hair cells of ksv/ksv mice had excessive growth. Hence, these data indicated that decreased MYO6 levels in ksv/ksv mutants disrupt actin networks in the apical region of hair cells, thereby maintaining the normal structure of the cuticular plates and rootlets, and additionally provided a cellular basis for stereociliary fusion in Myo6 mutants.

[Comparison of Contrast to Noise Ratio and Signal Difference to Noise Ratio Based on QA and QC Guidelines in CR Mammography].

The measurement methods of contrast to noise ratio (CNR) and signal difference to noise ratio (SDNR) in digital mammography are different among several quality assurance (QA) guidelines, that is, the type of pixel value (PV), phantom shape, location of aluminum plate, and the size of region of interest (ROI) principally differ in data acquisition. We compared CNR (SDNR) obtained from three QA guidelines. They are the European Reference Organisation for Quality Assured Breast Screening and Diagnostic Services (EUREF), the International Electrotechnical Commission (IEC), and the International Atomic Energy Agency (IAEA). In EUREF and IEC, CNR was calculated using linearized pixel value (LPV). In IAEA, because the type of pixel value to use in SDNR was not specified, SDNR was calculated using PV and LPV, and CNR was calculated using LPV. Target/filter combinations are molybdenum/molybdenum (Mo/Mo) and molybdenum/rhodium (Mo/Rh). Applied various tube voltages are 25, 30, and 35 kV, and various phantom thicknesses are 20, 45, and 70 mm of polymethyl methacrylate (PMMA). The PV-SDNR of IAEA showed the largest value among the three methods, following LPV-CNR of IEC, LPV-CNR of EUREF at 20 mm PMMA thickness. In IAEA, SDNR changed by the kind of pixel value (PV or LPV). When CNR is calculated, every researcher should describe the type of guidelines, the kind of pixel value, and formula for calculation.

[Influence of Detector Radiation Damage on CR Mammography Quality Control].

Recently, radiation damage to the detector apparatus employed in computed radiography (CR) mammography has become problematic. The CR system and the imaging plate (IP) applied to quality control (QC) program were also used in clinical mammography in our hospital, and the IP to which radiation damage has occurred was used for approximately 5 years (approximately 13,000 exposures). We considered using previously acquired QC image data, which is stored in a server, to investigate the influence of radiation damage to an IP. The mammography unit employed in this study was a phase contrast mammography (PCM) Mermaid (KONICA MINOLTA) system. The QC image was made newly, and it was output in the film, and thereafter the optical density of the step-phantom image was measured. An input (digital value)-output (optical density) conversion curve was plotted using the obtained data. The digital values were then converted to optical density values using a reference optical density vs. digital value curve. When a high radiation dose was applied directly, radiation damage occurred at a position on the IP where no object was present. Daily QC for mammography is conducted using an American College of Radiology (ACR) accreditation phantom and acrylic disc, and an environmental background density measurement is performed as one of the management indexes. In this study, the radiation damage sustained by the acrylic disc was shown to differ from that of the background. Thus, it was revealed that QC results are influenced by radiation damage.

[Development of Determination Method of Ipfencarbazone in Agricultural Products, Livestock Products and Seafood by LC-MS/MS].

A method for the determination of ipfencarbazone in agricultural products, livestock products and seafood by LC-MS/MS was developed. Agricultural samples were extracted with acetone. An aliquot of crude extract was partitioned with n-hexane and sat. sodium chloride solution. Clean-up was performed using GC/PSA and C18 cartridges. In the case of livestock products and seafood, samples were extracted with a mixture of acetone and n-hexane, and the organic layer was collected. After acetonitrile-hexane partitioning, the extract was cleaned up using PAS and C18 cartridges. The gradient LC separation was performed on a C18 column with acetonitrile-water containing acetic acid as a mobile phase, and MS with positive ion electrospray ionization was used for detection. The average recoveries (n=5) of ipfencarbazone from 16 kinds of agricultural products, livestock products and seafood spiked at the MRLs or at the uniform limits (0.01 ppm) were 73-101%, and the relative standard deviations were 1.3-5.1%. The limit of quantitation of the developed method was 0.01 mg/kg for ipfencarbazone.

Putative Epimutagens in Maternal Peripheral and Cord Blood Samples Identified Using Human Induced Pluripotent Stem Cells.

The regulation of transcription and genome stability by epigenetic systems are crucial for the proper development of mammalian embryos. Chemicals that disturb epigenetic systems are termed epimutagens. We previously performed chemical screening that focused on heterochromatin formation and DNA methylation status in mouse embryonic stem cells and identified five epimutagens: diethyl phosphate (DEP), mercury (Hg), cotinine, selenium (Se), and octachlorodipropyl ether (S-421). Here, we used human induced pluripotent stem cells (hiPSCs) to confirm the effects of 20 chemicals, including the five epimutagens, detected at low concentrations in maternal peripheral and cord blood samples. Of note, these individual chemicals did not exhibit epimutagenic activity in hiPSCs. However, because the fetal environment contains various chemicals, we evaluated the effects of combined exposure to chemicals (DEP, Hg, cotinine, Se, and S-421) on hiPSCs. The combined exposure caused a decrease in the number of heterochromatin signals and aberrant DNA methylation status at multiple gene loci in hiPSCs. The combined exposure also affected embryoid body formation and neural differentiation from hiPSCs. Therefore, DEP, Hg, cotinine, Se, and S-421 were defined as an "epimutagen combination" that is effective at low concentrations as detected in maternal peripheral and cord blood.

Urethral caruncle in a 9-year-old girl: a case report and review of the literature.

INTRODUCTION: Urethral caruncles are the most frequent benign tumors of the female urethra. Most of them are found in post-menopausal women, and they are rare in childhood. Only a few pediatric cases have been published in the literature. In this report, we present an unusual case of a pediatric patient with a urethral caruncle, along with a review of the literature. CASE PRESENTATION: A 9-year-old Mongolian girl was referred to our hospital with a 2-week history of frequent adherence of a small amount of blood to her underwear. We found a sessile smooth margin, a clear boundary and an elastic, soft red tumor over the entire circumference of the urethral meatus. At the beginning, because of the child's age, urethral prolapse was suspected. There was no response after 3 weeks of conservative treatment with steroid ointment. With the patient under general anesthesia, a partial tumor resection was performed for the purpose of histological examination. The tumor excision was limited to about 1/2 laps of the urethral meatus to prevent the development of urethral stricture. On the basis of clinical and histopathological examinations, a diagnosis of a urethral caruncle was made. Post-operatively, steroid ointment application to residual masses was continued, and these disappeared about 6 months later. Our patient was free of recurrence and had had no complications after 3 years of follow-up. CONCLUSIONS: Urethral caruncles are rare in children, and the possibility of malignancy is slight during this period. Biopsy of the mass is not required for diagnosis. It should be indicated only if the mass has other characteristics that raise suspicion of malignancy. In previously reported cases, all of the tumor was removed. However, the trigger of the caruncle in childhood is chronic inflammation. Conservative therapy with steroid ointment should be the core treatment. However, it may be necessary to proceed to treatment because caruncles take a long time to heal. The case that we describe in this report will serve as an example for similar cases in the future.