The Influence of Polyploidy and Genome Composition on Genomic Imprinting in Mice

Genomic imprinting is an epigenetic mechanism that switches the expression of imprinted genes involved in normal embryonic growth and development in a parent-of-origin-specific manner. Changes inDNAmethylation statuses from polyploidization are a well characterized epigenetic modification in plants. However, how changes in ploidy affect both imprinted gene expression and methylation status in mammals remains unclear. To address this, we used quantitative real time PCR to analyze expression levels of imprinted genes in mouse tetraploid fetuses. We used bisulfite sequencing to assess the methylation statuses of differentially methylated regions (DMRs) that regulate imprinted gene expression in triploid and tetraploid fetuses. The nine imprinted genes H19, Gtl2, Dlk1, Igf2r, Grb10, Zim1, Peg3, Ndn, and Ipw were all unregulated; in particular, the expression of Zim1 was more than 10-fold higher, and the expression of Ipw was repressed in tetraploid fetuses. The methylation statuses of four DMRs H19, intergenic (IG), Igf2r, and Snrpn in tetraploid and triploid fetuses were similar to those in diploid fetuses. We also performed allele-specific RT-PCR sequencing to determine the alleles expressing the three imprinted genes Igf2, Gtl2, and Dlk1 in tetraploid fetuses. These three imprinted genes showed monoallelic expression in a parent-of-origin-specific manner. Expression of non-imprinted genes regulating neural cell development significantly decreased in tetraploid fetuses, which might have been associated with unregulated imprinted gene expression. This study provides the first detailed analysis of genomic imprinting in tetraploid fetuses, suggesting that imprinted gene expression is disrupted, but DNA methylation statuses of DMRs are stable following changes in ploidy in mammals

The Influence of Hyperactivity of the Hypothalamic-pituitary-adrenal Axis and Hyperglycemia on the 5-HT2A Receptor-mediated Wet-dog Shake Responses in Rats

Hyperactivity of the hypothalamic-pituitary-adrenal (HPA) axis induces hyperglycemia and serotonin (5-HT)2A receptor supersensitivity. In the present study, to investigate the effect of hyperglycemia on the function of 5-HT2A receptors, we compared the 5-HT2A receptor-mediated wet-dog shake responses in rats treated with adrenocorticotropic hormone (ACTH), dexamethasone and streptozotocin. ACTH (100 &#956;g/rat per day, s.c.), dexamethasone (1 mg/kg per day, s.c.) and streptozotocin (60 mg/kg, i.p.) produced significant hyperglycemia at 14 days after the start of these treatments, and the hyperglycemia was most pronounced in the streptozotocin-treated rats. The wet-dog shake responses induced by (±)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI), a 5-HT2A receptor agonist, were significantly enhanced at 14 days after repeated treatment with ACTH and dexamethasone. However, streptozotocin-induced diabetes had no effect on the wet-dog shake responses. The results of the present study suggest that hyperglycemia is not strongly associated with the enhanced susceptibility of 5-HT2A receptors under the condition of hyperactivity of the HPA axis.</p

Effects of Physical and Psychological Stress on 5-HT2A Receptor-mediated Wet-dog Shake Responses in Streptozotocin-induced Diabetic Rats.

Several epidemiological and clinical studies have indicated that the prevalence of psychiatric disorders is higher in diabetic patients than in the general population. In the present studies, we examined the behavioral changes in streptozotocin-induced diabetic rats, and investigated the effects of physical and psychological stress on the hippocampal BDNF levels and on the serotonin 2A (5-HT2A) receptor-mediated wet-dog shake responses. The streptozotocin (60 mg/kg, i.p.)-induced diabetes had no significant effects on the immobility time in the forced swim test or on locomotor activity in the open-field test. Moreover, there was no significant difference in the wet-dog shake responses induced by DOI, a 5-HT2A receptor agonist, between nondiabetic and diabetic rats. Five-day exposure to physical (electric footshock) and psychological (non-footshock) stress had no signifi cant effect on the hippocampal BDNF level in diabetic or nondiabetic rats. The 2 types of stress had no significant effect on the DOI-induced wet-dog shake responses in nondiabetic rats. In diabetic rats, the repeated exposure to physical stress markedly increased the DOI-induced wet-dog shake responses, but the repeated exposure to psychological stress had no effect. These results suggest that exposure to physical stress augmented the susceptibility to the wet-dog shake responses to 5-HT2A receptor stimulation in streptozotocin-induced diabetic rats

DRUG-ADMINISTERING PERSONS' EXPOSURE TO ORAL ANTICANCER DRUGS TO BE ADMINISTERED THROUGH A TUBE

Objective: The objective of this study was to quantitatively evaluate anticancer drug exposure of non-health care professionals who administer drugs through a tube employing a method devised by us. Methods: The subjects were 30 general volunteers aged 22-84 years. They wore gloves and administered Indian ink, simulating an anticancer drug, to a multipurpose adult human-type patient care simulator through a tube using 5 types of syringe, and the area stained with Indian ink was measured. Results: When comparing the number of pixels among the syringes regardless of age, Syringe B showed the lowest number (11.8±3.1 cm2), and there was a significant difference between Syringes B and E. Furthermore, we compared the total number of pixels in each age group regardless of the type of syringe. In the 20-year-old group, it was the lowest (10.9±2.3 cm2) showing significant differences in comparison with the other groups. When Syringe B was used, the number of pixels was markedly lower than on adopting the other syringes. Conclusion: It was clarified that the level of exposure to anticancer drugs markedly varies depending on the type of syringe and age. It was also clarified that the method to evaluate exposure to anticancer drugs using Indian ink devised by us is simple and useful

DRUG-ADMINISTERING PERSONS' EXPOSURE TO ORAL ANTICANCER DRUGS TO BE ADMINISTERED THROUGH A TUBE

ABSTRACTObjective: The objective of this study was to quantitatively evaluate anticancer drug exposure of non-health care professionals who administer drugsthrough a tube employing a method devised by us.Methods: The subjects were 30 general volunteers aged 22-84 years. They wore gloves and administered Indian ink, simulating an anticancer drug, toa multipurpose adult human-type patient care simulator through a tube using 5 types of syringe, and the area stained with Indian ink was measured.Results: When comparing the number of pixels among the syringes regardless of age, Syringe B showed the lowest number (11.8±3.1 cm2), and therewas a significant difference between Syringes B and E. Furthermore, we compared the total number of pixels in each age group regardless of the typeof syringe. In the 20-year-old group, it was the lowest (10.9±2.3 cm2) showing significant differences in comparison with the other groups. WhenSyringe B was used, the number of pixels was markedly lower than on adopting the other syringes.Conclusion: It was clarified that the level of exposure to anticancer drugs markedly varies depending on the type of syringe and age. It was alsoclarified that the method to evaluate exposure to anticancer drugs using Indian ink devised by us is simple and useful.Keywords: Oral anticancer drugs, Simple suspension method, Drug-administering persons' exposure

Prevention of Image Quality Degradation in Wider Field Optical Coherence Tomography Angiography Images Via Image Averaging

Purpose: To evaluate the mutual effect of widening the field of view and multiple en face image averaging on the quality of optical coherence tomography angiography (OCTA) images. Methods: This prospective, observational, cross-sectional case series included 20 eyes of 20 healthy volunteers with no history of ocular or systemic disease. OCTA imaging of a 3 × 3-mm, 6 × 6-mm, and 12 × 12-mm area centered on the fovea was performed nine times using the PLEX Elite 9000. We acquired averaged OCTA images generated from nine en face OCTA images. The corresponding areas in the three scan sizes were evaluated for the original single-scanned OCTA images and averaged OCTA images both qualitatively and quantitatively. Quantitative measurements included vessel density (VD), vessel length density (VLD), fractal dimension (FD), and contrast-to-noise ratio (CNR). Results: Significant differences in VD, VLD, FD, and CNR (P < 0.001) were observed due to the mutual effect of averaging and differences in scan size. Both qualitative and quantitative evaluations indicated that the quality of 6 × 6-mm averaged images was equal to or better than that of 3 × 3-mm single-scanned images. However, the quality of 12 × 12-mm averaged images did not reach that of 3 × 3-mm single-scanned images. Conclusions: To some extent, multiple en face OCTA image averaging can compensate for the deterioration in image quality caused by widening the field of view. Translational Relevance: Multiple en face OCTA image averaging can be a technique for acquiring wider field OCTA images with good quality

Trials for Risk Assessment of Japanese Encephalitis Based on Serologic Surveys of Wild Animals

Flavivirus Encephaliti