Inherited retinal degenerations (IRDs) are a group of phenotypically and genotypically heterogeneous disorders with substantial socioeconomic impact. In this cohort study, we tried to address the genetic characteristics and epidemiology of IRDs in Taiwan. Totally, 312 families with IRDs were identified and recruited and genetic testing was performed via probe capture-based NGS targeting 212 IRD-related genes. Statistical analysis was based on the proband of each affected family. Disease-causing genotypes were identified in 178 families (57.1%). ABCA4 variants were the most common cause of disease in this cohort (27 families, 15.2%), whereas CYP4V2 variants were the most common cause for the single phenotype—Bietti’s crystalline dystrophy (12 families, 3.8%). Some variants such as ABCA4:c.1804C>T, CYP4V2:c.802-8_810delinsGC, and EYS:c6416G>A were population-specific disease-causing hotspots. Probands affected by ABCA4, RPGR, RP1L1, and CEP290 sought medical help earlier while patients affected by EYS and CYP4V2 visited our clinic at an older age. To evaluate the representativeness of our cohort in the genetic epidemiology of IRDs in Taiwan, our demographic data were compared with that of the total IRD population in Taiwan, obtained from the National Health Insurance Research Database. This is currently the largest-scale, comprehensive study investigating the genetic characteristics and epidemiology of IRD in Taiwan. These data could help patients and caregivers to adopt precision genomic medicine and novel gene therapies in near future.

Arterial pressure of each new breeding spontaneous Phase-1 hypertension (P1-HT) rat was recorded for 5 min by intravascular femoral artery catheter that served as a reference value prior to treatment. In the acute antihypertensive test, 0.36 g/kg Bwt of Plantago asiatica seed extract (PSE) was administered, via gavage feeding, to P1-HT rats, and the arterial pressures were continuously recorded for 1 h. The acute antihypertensive effects of PSE on P1-HT rats appeared within 15 min after PSE administration and lasted over 1 h with systolic pressure decreased 31.5 mmHg and diastolic pressure decreased 18.5 mmHg. The systolic pressure decreased 28 mmHg and diastolic pressure decreased 16 mmHg in P1-HT rats when simultaneously compared with verapamil hydrochloride (reference drug), whereas there were no significant differences in the pretreated reference values of acute PSE treatment and the untreated control. In the chronic test, P1-HT rats received 0.36 g/kg Bwt day of PSE or equal volume of water for 4 weeks via oral gavage, and the lower blood pressure tendencies of chronic PSE treatment were also found when compared with the controls. The antihypertensive values of PSE were also confirmed in spontaneously hypertensive rats (SHRs). Oral administration with PSE can effectively moderate blood pressure within an hour, while taking PSE daily can control the severity of hypertension, suggesting PSE is a potentially antihypertensive herb.

The profiling of germline Adaptive Immune Receptor Repertoire (AIRR), including T cell receptors (TR) and immunoglobulin (IG), is medically important but challenging due to high genetic diversity and complex recombination. While databases such as the international ImMunoGeneTics information system (IMGT) provide a valuable collection of AIRR allele sequences, they are far from complete, and many of them lack flanking sequences outside alleles. It will be beneficial to have technologies that can efficiently capture relevant sequences from DNA samples or genome assemblies, perform accurate allele calling, and discover novel alleles. In this study, we developed gAIRR-seq, a probe capture-based targeted sequencing pipeline, to profile genomic sequences of TR and IG from individual DNA samples. We then proposed computational pipelines — gAIRR-call and gAIRR-annotate — to call alleles from gAIRR-seq reads and validate the results with whole-genome assemblies. Genomic DNA samples from Genome in a Bottle (GIAB) reference materials and public whole-genome assemblies were used for benchmarking. We applied gAIRR-call to genotype TRV and TRJ alleles with 100% accuracy and achieved 98% accuracy when considering flanking regions of V alleles. We used gAIRR-annotate to annotate allele positions and collect alleles and flanking regions into databases. gAIRR-annotate validated the alleles using 13 high-quality whole-genome assemblies on 6 samples and discovered 79 novel TRV alleles and 11 novel TRJ alleles. We provided a summary of the number of TR alleles from 13 high-quality draft assemblies with the proposed pipeline. We validated a 65-kbp and a 10-kbp structural variant for HG002 on chromosomes 7 and 14, where TRD and J alleles reside. We also uncovered the disagreement of the human genome GRCh37 and GRCh38 in the TR regions; GRCh37 possesses a 270 kbp inversion and a 10 kbp deletion in chromosome 7 relative to GRCh38.

Hereditary optic neuropathy (HON) is a group of genetically heterogeneous diseases that cause optic nerve atrophy and lead to substantial visual impairment. HON may present with optic nerve atrophy only or in association with various systemic abnormalities. Although a genetic survey is indispensable for diagnosing HON, conventional sequencing techniques could render its diagnosis challenging. In this study, we attempted to explore the genetic background of patients with HON in Taiwan through capture-based next-generation sequencing targeting 52 HON-related genes. In total, 57 patients from 48 families were recruited, with 6 patients diagnosed as having Leber hereditary optic neuropathy through initial screening for three common variants (m.3460G>A, m.11778G>A, m.14484T>C). Disease-causing genotypes were identified in 14 (33.3%) probands, and OPA1 variants were the most prevalent cause of autosomal HON. Exposure to medications such as ethambutol could trigger an attack of autosomal dominant optic atrophy. WFS1 variants were identified in three probands with variable clinical features in our cohort. Hearing impairment could occur in patients with OPA1 or WFS1 variants. This is the first comprehensive study investigating the genetic characteristics of HON in Taiwan, especially for autosomal HON. Our results could provide useful information for clinical diagnosis and genetic counseling in this field.

Introduction and hypothesisThe degeneration and lost on dopaminergic neurons in midbrain may cause Parkinson's disease, and it is one of the most common neurodegenerative disease in human. Rotenone is a natural lipophilic toxic compound that is easy to cross the blood‐brain barrier, and widely used as insecticide and pesticide. Rotenone may impair mitochondrial oxidative phosphorylation and lead to dopaminergic neuron loss. In this study we investigated the protective effects of caffeine on rotenone‐induced Parkinson's disease models in vitro and in vivo.MethodsIn vitro, the dopaminergic cell line, SH‐SY5Y cells, is pretreated with caffeine (0~100 mM) for 30 mins and followed by treatment with rotenone (0~10 mM) for 1 hr~24 hr respectively. The cell survival rate, proliferation and morphology were analysis by flow cytometry, ELISA, Cell Counting Kit‐8(CCK‐8) and histology studies. Cell apoptosis/necrosis were analyzed by Annexin‐V/PI staining using flow cytometry. In vivo, the SD rats pretreated with caffeine (10 mg/kg daily, P.O.) or saline and followed treated with rotenone (1 mg/kg daily for 1 week, S.C.). The expressions of apoptosis and autophagy related proteins were analyzed by western blotting. The locomotion of rats were carried out by open field tests.ResultsCaffeine treatments (0–100 mM) had rarely toxic effect on SH‐SY5Y cell lines but could promote autophagy. The SH‐SY5Y cells treated with 1 nM rotenone for 8 hr may decrease the mitochondrial activity of around 60%. Pretreated with caffeine (>2 mM) for 30 min and followed treated with 10 nM rotenone for 8 hr could significantly increase total cell numbers and decrease the cell apoptotic ratio, but no effect on the necrosis ratio and did not restore the mitochondrial activity on rotenone‐induced dopaminergic cell toxicity. The locomotion of SD rats that pretreated with caffeine was better than the saline control in rotenone‐induced Parkinson's disease model. After exposure to rotenone, the situation of recovery by bradykinesia in daily caffeine‐treated group is more obviously than saline control.ConclusionsIn rotenone‐induced Parkinson's disease model, caffeine could facilitate autophagy, increase cell viability and animal locomotion. Our results indicated that caffeine has the potential protective effects on rotenone‐induced Parkinson's disease model.Support or Funding InformationKMU‐TP105E37This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.