Quantitative DNA Methylation Analysis and Epigenotype-Phenotype Correlations in Taiwanese Patients with Silver-Russell Syndrome.

Background: Silver-Russell syndrome (SRS; OMIM #180860) is a clinically and genetically heterogeneous imprinting disorder characterized by prenatal and postnatal growth failure. The aim of this study was to identify the epigenotype-phenotype correlations in these patients using quantitative DNA methylation analysis. Methods: One hundred and eighty-three subjects clinically suspected of having SRS were referred for diagnostic testing by the methylation profiling of H19-associated imprinting center (IC) 1 and imprinted PEG1/MEST regions using methylation-specific high-resolution melting analysis and methylation quantification with the MassARRAY assay. Correlations between quantitative DNA methylation status and clinical manifestations of the subjects according to the Netchine-Harbison (N-H) clinical scoring system for SRS were analyzed. Results: Among the 183 subjects, 90 had a clinical diagnosis of SRS [N-H score ≥ 4 (maximum = 6)] and 93 had an SRS score < 4. Molecular lesions were detected in 41% (37/90) of the subjects with a clinical diagnosis of SRS, compared with 3% (3/93) of those with an N-H score < 4. The IC1 methylation level was negatively correlated with the N-H score. The molecular diagnosis rate was positively correlated with the N-H score. Thirty-one subjects had IC1 hypomethylation (IC1 methylation level <35% by the MassARRAY assay), seven had maternal uniparental disomy 7, and two had pathogenic copy number variants. Among the 90 subjects with an N-H score ≥ 4, the IC1 methylation level was significantly different between those with or without some clinical SRS features, including birth length ≤ 10th centile, relative macrocephaly at birth, normal cognitive development, body asymmetry, clinodactyly of the fifth finger, and genital abnormalities. Conclusions: This study confirmed the suitability of the N-H clinical scoring system as clinical diagnostic criteria for SRS. Quantitative DNA methylation analysis using the MassARRAY assay can improve the detection of epigenotype-phenotype correlations, further promoting better genetic counseling and multidisciplinary management for these patients.

Ciproxifan, a histamine H3 receptor antagonist and inverse agonist, presynaptically inhibits glutamate release in rat hippocampus.

Ciproxifan is an H3 receptor antagonist and inverse agonist with antipsychotic effects in several preclinical models; its effect on glutamate release has been investigated in the rat hippocampus. In a synaptosomal preparation, ciproxifan reduced 4-aminopyridine (4-AP)-evoked Ca2+-dependent glutamate release and cytosolic Ca2+ concentration elevation but did not affect the membrane potential. The inhibitory effect of ciproxifan on 4-AP-evoked glutamate release was prevented by the Gi/Go-protein inhibitor pertussis toxin and Cav2.2 (N-type) and Cav2.1 (P/Q-type) channel blocker ω-conotoxin MVIIC, but was not affected by the intracellular Ca2+-release inhibitors dantrolene and CGP37157. Furthermore, the phospholipase A2 (PLA2) inhibitor OBAA, prostaglandin E2 (PGE2), PGE2 subtype 2 (EP2) receptor antagonist PF04418948, and extracellular signal-regulated kinase (ERK) inhibitor FR180204 eliminated the inhibitory effect of ciproxifan on glutamate release. Ciproxifan reduced the 4-AP-evoked phosphorylation of ERK and synapsin I, a presynaptic target of ERK. The ciproxifan-mediated inhibition of glutamate release was prevented in synaptosomes from synapsin I-deficient mice. Moreover, ciproxifan reduced the frequency of miniature excitatory postsynaptic currents without affecting their amplitude in hippocampal slices. Our data suggest that ciproxifan, acting through the blockade of Gi/Go protein-coupled H3 receptors present on hippocampal nerve terminals, reduces voltage-dependent Ca2+ entry by diminishing PLA2/PGE2/EP2 receptor pathway, which subsequently suppresses the ERK/synapsin I cascade to decrease the evoked glutamate release.

Epigenotype, genotype, and phenotype analysis of patients in Taiwan with Beckwith-Wiedemann syndrome.

BACKGROUND: Beckwith-Wiedemann syndrome (BWS) is a congenital overgrowth disorder predisposing to tumorigenesis that results from abnormal expression or function of imprinted genes of chromosome 11p15.5. METHODS: Forty-seven patients in Taiwan with clinical suspicion of BWS were referred for diagnostic testing based on methylation profiling of H19-associated imprinting center (IC) 1 and KCNQ1OT1-associated IC2 using high-resolution melting analysis, multiplex ligation-dependent probe amplification, or high-resolution quantitative methylation profiling. RESULTS: Twenty-eight patients received a clinical diagnosis of BWS (the presence of 3 major features or 2 major features and at least 1 minor feature), 18 had suspected BWS (the presence of at least 1 major feature), and 1 had isolated Wilms' tumor. Nineteen patients were identified with IC2 hypomethylation (including 1 with isolated Wilms' tumor), 1 with IC1 hypermethylation, 2 with paternal uniparental disomy, and 1 with CDKN1C mutation. Several clinical features were found to be statistically different (P<0.05) between the 2 groups-clinical diagnosis of BWS (n=28) or suspected BWS (n=18)-including macroglossia, pre- or postnatal gigantism, abdominal wall defect, ear creases, facial nevus flammeus, BWS score, and the molecular diagnosis rate. Molecular lesion was detected in 81% of patients with the presence of three major features, compared with 33% and 28% of those with two or one major feature, respectively. The mean BWS score was 5.6 for 19 subjects with "IC2 hypomethylation", compared with 3.8 for 2 subjects with pUPD. The BWS score of one subject with CDKN1C mutation and one with IC1 hypermethylation was 6 and 7, respectively. CONCLUSIONS: The BWS score was positively correlated with the molecular diagnosis rate (P<0.01). The BWS database of epigenotype, genotype, and phenotype is expected to promote better genetic counseling and medical care of these patients.

Cardiac structure and function and effects of enzyme replacement therapy in patients with mucopolysaccharidoses I, II, IVA and VI.

BACKGROUND: While enzyme replacement therapy (ERT) has been shown to improve endurance and joint mobility for patients with mucopolysaccharidoses (MPS) I, II, IVA and VI, the impact of ERT on cardiac abnormalities remains uncertain. METHODS: Medical records and echocardiograms of 28 Taiwanese MPS patients (9 with MPS I, 7 with MPS II, 7 with MPS IVA, and 5 with MPS VI) treated with ERT for 1-10.8years were retrospectively reviewed. RESULTS: At start of ERT, z scores>2 were identified in 46% and 75% for left ventricular mass index (LVMI) and interventricular septum thickness in diastole (IVSd) in these patients, respectively. Twenty-four patients (86%) had valvular heart disease. After ERT, the mean IVSd z score of all patients decreased significantly from 3.87 to 2.57 (p=0.016). For 11 patients starting ERT before 12years of age, z scores for both LVMI and IVSd decreased significantly (p<0.01) after ERT. However, the condition of valve regurgitation or stenosis did not show improvement despite ERT. CONCLUSIONS: ERT was shown to be an effective therapy for reducing cardiac hypertrophy, with best results seen when ERT was started at an early age. ERT, however, had little impact on valvular heart disease.

Generalized bullous fixed drug eruption is distinct from Stevens-Johnson syndrome/toxic epidermal necrolysis by immunohistopathological features.

BACKGROUND: Generalized bullous fixed drug eruption (GBFDE), a particular form of fixed drug eruption (FDE), is characterized by widespread blisters and erosions and can be confused with Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN). OBJECTIVE: We sought to analyze specific features of GBFDE and differentiate it from SJS/TEN. METHODS: We retrospectively studied patients with GBFDE and SJS/TEN during a period of 10 years. GBFDE was defined as typical FDE lesions with blisters involving at least 10% body surface area on at least 3 of 6 different anatomic sites. Clinical presentations; histopathological features; immunohistochemical patterns of cluster-of-differentiation (CD)3, CD4, CD8, CD56, Fas, Fas ligand, granzyme B, perforin, granulysin, and forkhead box P3 (Foxp3); and serum granulysin levels were compared. RESULTS: Twenty-three cases of GBFDE were collected. Patients with GBFDE had shorter latent periods, less mucosal involvement, more eosinophil infiltration, and dermal melanophages. Lesional infiltrates in GBFDE had more dermal CD4(+) cells including Foxp3(+) regulatory T cells, fewer intraepidermal CD56(+) cells, and fewer intraepidermal granulysin(+) cells. The serum level of granulysin in GBFDE was also significantly lower than in SJS/TEN. LIMITATIONS: The number of cases in this study is small. CONCLUSION: GBFDE is a distinct disease distinguishable from SJS/TEN by particular features such as granulysin, CD56, and Foxp3 expressions.

Monitoring ambient air pollutants and apply Woods' model in the prediction seasonal dry deposition at Chang-Hua (urban) and Kao-Mei (wetland) county, Taiwan.

The main purpose for this study was to monitor ambient air particles and metallic elements (Mn, Fe, Zn, Cr, Cu and Pb) in total suspended particulate (TSP) concentration and dry deposition. In addition, the calculated/measured dry deposition flux ratios of ambient air particles and metallic elements (Mn, Fe, Zn, Cr, Cu and Pb) were evaluated using Woods' model at urban and wetland areas for the 2009-2010 period. The results indicated that the mean highest concentrations of metallic elements Mn, Fe, Zn, Cr, Cu and Pb in TSP were found in Chang-Hua (urban) sampling site. And as for the two characteristic sampling sites, the Woods' model exhibits better dry deposition of particulates of 18 µm particle size than the rest of the other particle sizes at any sampling site in this study. The average calculated/measured flux ratios for two seasons (summer and fall) by using Woods model at 2.5, 10 and 18 µm particles sizes were also studied. The results indicated that the average calculated/measured flux ratios orders for two seasons of various particles sizes were all displayed as Fe > Mn > Zn > Cu > Cr > Pb > particle. And these calculated/measured flux ratios orders were Fe > Mn > Cu > Zn > Cr > Pb > particle and were Fe > Mn > Zn > Cu > Cr > particle > Pb, during spring and winter seasons, respectively. Finally, in the spring and summer seasons of Gao-Mei (wetland) sampling site, the average calculated/measured flux ratios using Woods' model was found to be 2.5, 10 and 18 µm, showing the order of the calculated/measured flux ratios to be Fe > Cu > Zn > Mn > Cr > Pb > particle. And the calculated/measured flux ratio orders were Fe > Zn > Mn > Cu > Cr > particle > Pb and were Fe > Cu > Zn > Mn > Cr > particle > Pb for fall and winter season, respectively.

Concentrations of particulates in ambient air, gaseous elementary mercury (GEM), and particulate-bound mercury (Hg(p)) at a traffic sampling site: a study of dry deposition in daytime and nighttime.

In this investigation, the concentrations of particles in ambient air, gaseous elemental mercury (GEM), and particulate-bound mercury (Hg(p)) in total suspended particulates (TSP) as well as dry deposition at a (Traffic) sampling site at Hung-kuang were studied during the day and night in 2012. The results reveal that the mean concentrations of TSP in ambient air, GEM, and Hg(p) were 69.72 µg/m(3), 3.17, and 0.024 ng/m(3), respectively, at the Hung-kuang (Traffic) sampling site during daytime sampling periods. The results also reveal that the mean rates of dry deposition of particles from ambient air and Hg(p) were 145.20 µg/m(2) min and 0.022 ng/m(2) min, respectively, at the Hung-kuang (Traffic) sampling site during the daytime sampling period. The mean concentrations of TSP in ambient air, GEM, and Hg(p) were 60.56 µg/m(3), 2.74, and 0.018 ng/m(3), respectively, at the Hung-kuang (Traffic) sampling site during the nighttime sampling period. The mean rates of dry deposition of particles and Hg(p) from ambient air were 132.58 µg/m(2) min and 0.016 ng/m(2) min, respectively, at the Hung-kuang (Traffic) sampling site during the nighttime sampling period.

Using deep learning to accurately detect sow vulva size in a group pen with a single camera.

This paper presents a non-contact method for the detection of changes in sow vulva size in a group pen. The traditional approach to estrus detection is manually pressing down on the back of the sow to elicit standing responses; however, this method causes undue distress for sows not in estrus. When a sow is in estrus, the vulva is red and swollen due to the presence of endocrine. Monitoring changes in vulva size to detect estrus with as little impact on the sow as possible is the focus of this study. This is achieved using a single camera combined with a deep learning framework. Our approach comprises two steps: vulva detection and vulva size conversion. Images of sows of Yorkshire, Landrace, and Duroc breeds were collected in group housing, and the vulva was detected through artificial markers and the network architecture of YOLO v4. Based on the internal and external parameters of the camera, the detected size was converted into millimeters and the results of manual measurement (MM) and automatic calculation combined to calculate the size of the vulva. Analysis of the calculated size compared with MM indicates that the object recognition rate of the system exceeds 97.06%, with a size error of only + 1.70 to -4.47 mm and high-calculation efficiency (>2.8 frames/s). Directions for future research include the automatic detection of pig width.

The size of a sow's vulva is an important indicator of sow estrus. Non-contact means of monitoring size changes for estrus timing would represent a significant contribution to the field of pig farming. This paper thus focuses on development of a system for the automatic detection of sow vulva size using a single camera combined with a deep learning framework. Experiments showed that the object recognition rate of the system exceeds 97.06%, the vulva size error is +1.70 to −4.47 mm, and the calculation efficiency is high (>2.8 frames/s).