DAJIN enables multiplex genotyping to simultaneously validate intended and unintended target genome editing outcomes.

Genome editing can introduce designed mutations into a target genomic site. Recent research has revealed that it can also induce various unintended events such as structural variations, small indels, and substitutions at, and in some cases, away from the target site. These rearrangements may result in confounding phenotypes in biomedical research samples and cause a concern in clinical or agricultural applications. However, current genotyping methods do not allow a comprehensive analysis of diverse mutations for phasing and mosaic variant detection. Here, we developed a genotyping method with an on-target site analysis software named Determine Allele mutations and Judge Intended genotype by Nanopore sequencer (DAJIN) that can automatically identify and classify both intended and unintended diverse mutations, including point mutations, deletions, inversions, and cis double knock-in at single-nucleotide resolution. Our approach with DAJIN can handle approximately 100 samples under different editing conditions in a single run. With its high versatility, scalability, and convenience, DAJIN-assisted multiplex genotyping may become a new standard for validating genome editing outcomes.

Validation of dual atrioventricular nodal physiology in dual atrioventricular nodal non-reentrant tachycardia via adenosine triphosphate injection during atrial pacing: A novel insight into the role of leftward inferior extension.

INTRODUCTION: Dual atrioventricular nodal non-reentrant tachycardia (DAVNNT) is a rare and challenging-to-diagnose arrhythmia, without previous reports associating it with a leftward inferior extension (LIE). METHODS: Diagnosis was made using adenosine triphosphate (ATP) injection during atrial pacing in a suspected DAVNNT patient. RESULTS: Ablation of the rightward inferior extension was unsuccessful in eliminating DAVNNT; however, subsequent ablation of the LIE successfully eradicated the arrhythmia. CONCLUSION: This unique case, marked by the first instance of DAVNNT caused by LIE, diagnosed through ATP injection, underscores the utility of this diagnostic approach and broadens the spectrum of our understanding and management of this condition.

[Decreased appetite is associated with a decreased cognitive function in elderly patients with diabetes].

OBJECTIVE: To examine the relationship between decreased appetite and the cognitive function in elderly diabetic patients. METHODS: The study subjects were outpatients with diabetes who were 60 years of age or older, and who were managed at Ise Red Cross Hospital. The cognitive function was assessed using a self-administered Dementia Checklist. The Japanese version of the Simplified Nutritional Appetite Questionnaire (SNAQ) was used to measure decreased appetite. A logistic regression analysis, in which the dependent variable was cognitive decline and the explanatory variables were appetite loss and adjustment variables, was used to calculate the odds ratio for cognitive decline according to the presence of appetite loss. RESULTS: Four hundred eighty patients were included in the analysis. Seventeen percent of the patients had decreased appetite and 21% had a decreased cognitive function. The unadjusted and adjusted odds ratios of cognitive decline for those with decreased appetite were 2.78 (95% confidence interval (CI), 1.66-4.65; P<0.001) and 2.26 (95% CI, 1.16-4.37; P=0.015), respectively, based on the absence of decreased appetite. CONCLUSION: Decreased appetite in elderly patients with diabetes was associated with a decreased cognitive function.

[Relationship between the type and number of social participation events and the cognitive function in older diabetics].

OBJECTIVE: To examine the relationship between the type and number of social participation events and the cognitive function in elderly patients with diabetes. METHODS: The subjects were diabetic outpatients of ≥60 years of age who were managed at Ise Red Cross Hospital. Mild cognitive impairment (MCI) and dementia were assessed using a self-administered dementia checklist. Six types of social participation activities and the number of participants were investigated. A logistic regression analysis with MCI and dementia as dependent variables, social participation as an explanatory variable, and adjustment variables was used to calculate the odds ratios for social participation according to the presence of MCI and dementia. RESULTS: In total, 352 patients were included in the analysis. Volunteer activities (P=0.012), hobbies (P=0.006), activities to share skills and experiences (P=0.026), and work (P=0.003) were significantly associated with dementia. Regarding the association between the amount of social participation and dementia, there was a decrease in the risk of dementia when the number of social participation was 2. However, social participation was not significantly associated with MCI in this study. CONCLUSION: The type and number of social participation events were found to be associated with the risk of dementia in elderly patients with diabetes.

[Decreased family caregiver satisfaction with diabetes treatment of elderly patients is associated with patient depressive symptoms].

OBJECTIVE: To examine the relationship between the dissatisfaction of family caregivers with diabetes treatment and depressive symptoms among the elderly. METHODS: The subjects were diabetic patients of 65 years of age or older and their family caregivers, who were outpatients at Ise Red Cross Hospital. The Japanese version of the Patient Health Questionnaire 9, which consists of nine items, was used to measure depressive symptoms. The Japanese version of the Treatment Satisfaction Scale for Caregivers of Dependent Diabetic Patients (STCD2-J) was used to measure the satisfaction of family caregivers with diabetes treatment. A logistic regression analysis was performed using depressive symptoms as dependent variable, satisfaction of the family caregiver with diabetes treatment as an explanatory variable, and adjustment variables. RESULTS: In total, 272 patients were included in the analysis. Taking the quintiles of STCD2-J scores, the adjusted odds ratios for patient depressive symptoms in Q2 (27-29), Q3 (24-26), Q4 (22-23) and Q5 (14-21) based on Q1 (30-36) (the group with the highest STCD2-J scores) were 2.44 (95% confidence interval (CI), 0.69-8.61; P=0.163), 3.08 (95% CI, 0.93-10.12; P=0.063), 2.69 (95% CI, 0.68-10.65; P=0.156), and 4.54 (95% CI, 1.44-14.32; P=0.010), respectively. CONCLUSION: We found that family caregivers' decreased satisfaction with diabetes treatment was associated with depressive symptoms. It is important to alert primary care physicians about depressive symptoms when they see family caregivers who show decreased satisfaction with diabetes treatment.

[Vegetable-first eating habits are associated with a reduced risk of mild cognitive impairment in elderly diabetic patients].

OBJECTIVE: To examine the relationship between vegetable-first eating habits and the cognitive function in elderly patients with diabetes. METHODS: The subjects were outpatients with diabetes ≥60 years old at Ise Red Cross Hospital. A self-administered dementia checklist was used to assess mild cognitive impairment and dementia. The participants were asked to fill out a questionnaire on their vegetable-first eating habits and were classified into 4 groups: 0, 1, 2, and 3 times a day. The dependent variables were mild cognitive impairment and dementia, and the explanatory variable was vegetable-first eating habits (0 as a reference). RESULTS: In total, 358 patients were included in the analysis. The number of vegetable-first meals was 0 in 153 (42.7%), 1 in 48 (13.4%), 2 in 46 (12.8%), and 3 in 111 (31.1%) patients. The adjusted odds ratios for 1, 2, and 3 times of mild cognitive impairment were 0.83 (95% confidence interval [CI], 0.35-1.94; P=0.680), 0.81 (95% CI, 0.32-2.00; P=0.653), and 0.37 (95% CI, 0.17-0.81; P=0.014), respectively. However, there was no significant association between vegetable-first eating habits and dementia. CONCLUSION: In elderly patients with diabetes, a vegetable-first eating habit at each meal was associated with a decreased risk of mild cognitive impairment.

[Difficulty experienced by family in supporting elderly diabetic patients is associated with a decline in patients' higher-level functions].

OBJECTIVE: To examine the relationship between difficulties experienced by family in supporting elderly patients with diabetes and these patients' higher-level functions. METHODS: The subjects were outpatients with diabetes ≥65 years old at Ise Red Cross Hospital and their family members. The Tokyo Metropolitan Institute of Gerontology Index of Competence (TMIG-IC) was used to assess patients' higher-level functions. The Japanese version of the Diabetes Caregiver Activity and Support Scale (D-CASS-J) was used to measure difficulties experienced by family in supporting elderly patients with diabetes. Using a multiple regression analysis with TMIG-IC scores (instrumental ADL, intellectual activity, and social participation) as the dependent variable and D-CASS-J scores (based on the highest scoring Q1 group among the three quartiles of D-CASS-J scores) as the explanatory variables, standardized regression coefficients (ß) for higher-level functions on the family's perceived support difficulties were calculated. RESULTS: In total, 429 patients (254 male patients and 175 female patients) were included in the analysis. For male patients, the adjusted beta values for TMIG-IC scores in Q2 and Q3 were -0.039 (P=0.649) and -0.352 (P<0.001), respectively, and the adjusted beta values for the instrumental ADL scores were -0.064 (P=0.455), -0.192 (P=0.047), -0.090 (P=0.375), and -0.360 (P=0.002) for the Intellectually Active scores, respectively, and the adjusted beta for social role scores were 0.054 (P=0.581) and -0.261 (P=0.019), respectively. However, there was no association between the patients' higher-level functions and family support difficulties among female patients. CONCLUSIONS: Difficulty experienced by the family in supporting elderly male patients with diabetes is associated with reduced higher-level functioning.

Retrograde fast pathway cryoablation inside the coronary sinus for slow-fast atrioventricular nodal reentrant tachycardia in a patient with persistent left superior vena cava.

INTRODUCTION: Persistent left superior vena cava (PLSVC) is accompanied by enlarged coronary sinus (CS) and deformation of the triangle of Koch. This makes anatomical evaluation of the atrioventricular (AV) nodal pathways difficult. METHODS: We attempted cryoablation of retrograde fast pathway located in the enlarged CS roof of PLSVC for slow-fast AV nodal reentrant tachycardia (AVNRT) induced by inadvertent antegrade fast pathway elimination during ablation of left atrial tachycardia. RESULTS: Slow-fast AVNRT was successfully eliminated without AV block progression. CONCLUSIONS: This is the first case of successful retrograde fast pathway ablation of the CS ostial roof for slow-fast AVNRT with PLSVC.

How to perform effective cryoballooon ablation of the left atrial roof: Considerations after experiencing more than 1000 cases.

INTRODUCTION: Cryoballoon ablation (CBA) of the left atrial (LA) roof in addition to a pulmonary vein isolation has been expected to improve the clinical outcomes post-atrial fibrillation (AF) ablation. We demonstrated the characteristics and efficacy of CBA of the LA roof through our experience with a large volume of procedures. METHODS: Among 1036 AF ablation procedures with CBA of the LA roof, 834 patients who underwent a de novo ablation were analyzed. RESULTS: Complete LA roof line conduction block was obtained in 767 patients (92.0%) solely by CBA (Group A). Compared with the other patients (Group B), the mean nadir balloon temperature during CBA of the LA roof (-44.5 ± 5.6°C for Group A vs. -40.5 ± 7.5°C for Group B, p < .01) and number of cryoballoon applications during the LA roof ablation with a circular mapping catheter located in the left superior pulmonary vein (1.3 ± 0.8 for Group A vs. 1.6 ± 1.0 for Group B, p = .02) were significantly lower in Group A. A multivariate analysis revealed that those were predictors of a complete LA roof conduction block after only CBA. The 1-year Kaplan-Meier atrial arrhythmia free rate estimates were 80.6% for Group A and 59.0% for Group B (p < .01). CONCLUSION: Complete LA roof line conduction block could be obtained with a cryoballoon without touch-up ablation in most cases. The LA roof CBA with a circular mapping catheter located in the right superior pulmonary vein was preferable to obtaining complete LA roof conduction block, which was important with regard to the clinical outcomes.

Incidence of sarcopenic obesity in older patients with diabetes and association between sarcopenic obesity and higher-level functional capacity: evaluation based on a consensus statement.

We used a consensus statement to diagnose sarcopenic obesity, evaluated incidence of sarcopenic obesity in older patients with diabetes, and examined whether sarcopenic obesity was associated with their higher-level functional capacity. Outpatients with diabetes (age, ≥65 years) undergoing treatment at Ise Red Cross Hospital were included. The Tokyo Metropolitan Institute of Gerontology Index of Competence (TMIG-IC)-a self-administered questionnaire-was used to assess their higher-level functional capacity. Sarcopenic obesity was evaluated based on the consensus statement diagnostic criteria-i.e., presence or absence of decreased skeletal muscle mass was evaluated based on appendicular skeletal muscle mass/body weight and obesity was assessed based on body fat mass percentage. To calculate the adjusted ß coefficient of sarcopenic obesity for higher-level functional capacity, multiple regression analyses were performed using TMIG-IC scores as the dependent variable and four categories (non-sarcopenia/non-obesity was used as a reference) that included sarcopenia and obesity as the predictor and moderator variables. Among the 310 patients included, the sarcopenic obesity incidence was 13.1% and 14.2% in men and women, respectively. When the non-sarcopenia/non-obesity group was used as a reference, the adjusted ß coefficient of sarcopenic obesity for scores of the TMIG-IC was -2.09 (p = 0.014) in men. However, the women showed no relationship between sarcopenic obesity and TMIG-IC scores. In older men with diabetes, sarcopenic obesity was associated with a decline in higher-level functional capacity.

["Koshi-heso" (waist-umbilicus) test: A novel screening method for visceral fatty obesity].

OBJECTIVE: To evaluate the screening performance and validity of the "Koshi-heso" (waist-umbilicus) test for visceral fatty obesity in elderly diabetic patients. METHODS: Subjects were diabetic patients ≥65 years old visiting our outpatient clinic. As a "Koshi-heso" test, the distance between the umbilicus and the superior border of the iliac crest (waist) was measured with the patient's own finger. When the index finger reached the umbilicus and there was a gap between the finger and the abdominal wall, the patient was classified as "smaller"; when the index finger reached the umbilicus and there was no gap between the finger and the abdominal wall, the patient was classified as "just fit"; and when the index finger did not reach the umbilicus, the patient was classified as "bigger". Abdominal circumference was used to assess visceral fat obesity, with a cut-off value of ≥85 cm for men and ≥90 cm for women. Visceral fat mass and body fat percentage were evaluated by the multi-frequency bioelectrical impedance method. The sensitivity and specificity of the waist-umbilical test for visceral fat obesity were calculated. Pearson's correlation coefficients between the "Koshi-heso" test and visceral fat mass and body fat percentage were calculated to evaluate the validity of the test. Furthermore, the association between the "Koshi-heso" test and risk factors for vascular disease, microvascular complications and cardiovascular disease was assessed by a logistic analysis. RESULTS: A total of 221 patients were included in the analysis population of the study. The cut-off values of "just fit" in men (sensitivity 0.96, specificity 0.62) and "bigger" in women (sensitivity 0.76, specificity 0.78) were optimal. Furthermore, the "Koshi-heso" test was significantly correlated with abdominal visceral fat mass and body fat percentage as well as with vascular disease risk factors and microvascular complications. CONCLUSION: The "Koshi-heso" test was able to be used as a screening method for visceral fatty obesity in elderly diabetic patients.

[A low phase angle is a factor independently associated with falls in elderly diabetic patients].

OBJECTIVE: To examine the relationship between a low phase angle (PA) and falls in elderly diabetic patients. METHODS: The subjects were diabetic patients ≥65 years old who were outpatients at Ise Red Cross Hospital. Patients were asked about their fall history using a self-administered questionnaire. The PA was measured by the multi-frequency bioelectrical impedance method, and the subjects were classified into two groups: the first quartile with the smallest PA (T1 group) and the second and third quartiles (T2/3 groups). A logistic regression analysis with falls as the dependent variable, PA as the explanatory variable, and adjustment variables was used to calculate the odds ratio of the PA for falls. RESULTS: A total of 255 patients were included in the analysis of this study. Of these, 33.3% were in the T1 group, 66.7% were in the T2/3 group, and 28.2% had fallen. The unadjusted and adjusted odds ratios for falls in the PA were 2.92 (95% confidence interval [CI], 1.31-4.07; P=0.004) and 2.34 (95% CI, 1.07-5.09; P=0.031), respectively. CONCLUSION: A low PA was associated with falls in elderly diabetic patients.

[Loneliness is associated with the risk of low nutrition in elderly diabetic patients].

OBJECTIVE: The purpose of this study was to examine the relationship between loneliness and malnutrition in elderly diabetic patients. METHODS: The subjects were diabetic patients ≥65 years old who were outpatients at Ise Red Cross Hospital. The nutritional status was assessed using the Mini Nutritional Assessment Short Form (MNA-SF), and patients were defined as being undernourished if their total score was <11 points. Loneliness was assessed using the Japanese version of the short form of the loneliness scale, a self-administered questionnaire, and a total score of ≥6 was considered to indicate loneliness. A logistic regression analysis was used to calculate the adjusted odds ratio of loneliness to undernutrition, with the dependent variable being undernutrition, the explanatory variable being loneliness, and the adjustment variable being loneliness. RESULTS: A total of 163 patients were included in the analysis of this study. Of these, 25.8% were lonely, and 33.7% were undernourished. The unadjusted and adjusted odds ratios of loneliness to undernutrition were 2.55 (95% confidence interval [CI], 1.24-5.27; P=0.011) and 3.81 (95% CI, 1.27-11.39; P=0.017), respectively. CONCLUSION: Loneliness is associated with a low nutritional status in elderly diabetic patients. It is important to alert diabetic patients with loneliness to their low nutritional status when they are diagnosed.

[A study of the frequency of cachexia and its associated factors in elderly patients with diabetes mellitus].

OBJECTIVE: To investigate the frequency of cachexia and its associated factors in elderly diabetic patients. METHODS: The subjects were diabetic patients ≥65 years old attending the outpatient diabetes clinic of Ise Red Cross Hospital. Cachexia was evaluated as having three or more of the following: (1) muscle weakness, (2) fatigue, (3) anorexia, (4) decreased lean body mass, and (5) biochemical abnormalities. A logistic regression analysis was used to identify factors associated with cachexia, with the dependent variable as cachexia and explanatory variables as various variables (basic attributes, glucose-related parameters, comorbidities, and treatment). RESULTS: A total of 404 patients (233 males and 171 females) were included in the study. Twenty-two (9.4%) and twenty-two (12.8%) male and female patients, respectively, had cachexia. A logistic regression analysis showed that the HbA1c value (odds ratio [OR], 0.26,95% confidence interval [CI], 0.08-0.81; P=0.021) and cognitive and functional decline (OR, 11.81, 95% CI, 1.81-76.95; P = 0.010) were factors associated with cachexia. In women, type 1 diabetes (OR, 12.39, 95% CI, 2.33-65.87; P=0.003), the HbA1c value (OR, 1.71, 95% CI, 1.07-2.74; P=0.024), and insulin usage (OR, 0.14, 95% CI, 0.02-0.71; P=0.018) were cachexia-related factors. CONCLUSIONS: The frequency of cachexia in elderly diabetic patients and its associated factors were identified. It is important to increase awareness of the risk of cachexia in elderly diabetic patients with poor glycemic control, cognitive and functional decline, type 1 diabetes mellitus, and insulin non-use.

Efficacy of left atrial posterior wall isolation guided by lesion size prediction module for non-paroxysmal atrial fibrillation.

AIMS: The efficacy of left atrial posterior wall isolation (PWI) is controversial. Lesion durability may be a major cause of arrhythmia recurrence. The use of the lesion size prediction module improves lesion durability. This study aimed to compare the clinical outcomes of pulmonary vein isolation (PVI) alone and PWI in addition to PVI (PVI + PWI) in patients with non-paroxysmal atrial fibrillation (AF) using a lesion size prediction module. METHODS AND RESULTS: This study is a sub-analysis of the recently published prospective multicentre observational study called ALCOHOL-AF (association of alcohol consumption with outcome of catheter ablation of AF). In this sub-analysis, patients with non-paroxysmal AF in whom PVI alone or PVI + PWI was performed using the lesion size prediction module were included. Freedom from atrial tachyarrhythmia was compared between PVI alone and PVI + PWI groups using propensity score analyses. Of the 3474 patients registered in the ALCOHOL-AF study, 572 patients (age 65.6 ± 10.1 years, male 77.4%, longstanding persistent AF 25.5%) were included in this sub-analysis. We selected 212 patients treated with PVI alone and 212 treated with PVI + PWI using one-to-one propensity score matching. During the follow-up period, atrial tachyarrhythmia recurrence was documented in 92 (43.4%) and 50 (23.6%) patients in the PVI alone and PVI + PWI groups, respectively. Freedom from atrial tachyarrhythmia recurrence without anti-arrhythmic drugs after a single procedure was significantly higher in PVI + PWI than in PVI alone groups (hazard ratio: 0.452, 95% confidence interval: 0.308-0.664, P < 0.001). CONCLUSION: In this hypothesis-generating study, lesion size prediction module-guided PVI + PWI was associated with better clinical outcomes than PVI alone in patients with persistent or longstanding persistent AF.

L17ER4: A cell-permeable attenuated cationic amphiphilic lytic peptide.

We have developed a series of attenuated cationic amphiphilic lytic (ACAL) peptides that can efficiently bring immunoglobulin G (IgG) and other functional proteins into cells. Delivery is generally achieved through the coadministration of ACAL peptides with cargo proteins. However, conjugation of ACAL peptides with cargos may be a promising approach for in vivo application to link in vivo outcomes of ACAL peptides and cargos. This study describes the creation of a new cell-permeable ACAL peptide, L17ER4. L17E is an optimized prototype of ACAL peptides previously developed in our laboratory for efficient delivery of IgGs into cells. Delivery was improved by functionalizing L17E with a tetra-arginine (R4) tag. Compared to the use of R8, a representative cell-penetrating peptide with high intracellular delivery efficacy, conjugation with L17ER4 afforded approximately four-fold higher cellular uptake of model small-molecule cargos (fluorescein isothiocyanate and HiBiT peptide). L17ER4 was also able to deliver proteins to cells. Fused with L17ER4, Cre recombinase was delivered into cells. Intracerebroventricular injection of Cre-L17ER4 into green red reporter mice, R26GRR, led to significant in vivo gene recombination in ependymal cells, suggesting that L17ER4 may be used as a cell-penetrating peptide for delivering protein therapeutics into cells in vivo.

Efficient production of large deletion and gene fragment knock-in mice mediated by genome editing with Cas9-mouse Cdt1 in mouse zygotes.

Genetically modified mouse models are essential for in vivo investigation of gene function and human disease research. Targeted mutations can be introduced into mouse embryos using genome editing technology such as CRISPR-Cas. Although mice with small indel mutations can be produced, the production of mice carrying large deletions or gene fragment knock-in alleles remains inefficient. We introduced the nuclear localisation property of Cdt1 protein into the CRISPR-Cas system for efficient production of genetically engineered mice. Mouse Cdt1-connected Cas9 (Cas9-mC) was present in the nucleus of HEK293T cells and mouse embryos. Cas9-mC induced a bi-allelic full deletion of Dmd, GC-rich fragment knock-in, and floxed allele knock-in with high efficiency compared to standard Cas9. These results indicate that Cas9-mC is a useful tool for producing mouse models carrying targeted mutations.

[Loneliness is associated with a reduced higher-level-function capacity in older diabetic patients].

OBJECTIVE: The present study examined the relationship between loneliness and higher-level functions in elderly diabetic patients. METHODS: The subjects were diabetic patients ≥65 years old who were outpatients at Ise Red Cross Hospital. The Tokyo Metropolitan Institute of Gerontology Index of Competence (TMIG-IC), a self-administered questionnaire, was used to assess higher-level functions. To evaluate loneliness, we used a shortened version of the Japanese version of the loneliness scale, a self-administered questionnaire. A multiple regression analysis with TMIG-IC scores as the dependent variable, loneliness as the explanatory variable, and adjustment variables was used to calculate the adjusted partial regression coefficients of loneliness on higher-level functions. RESULTS: One hundred and seventy patients were included in the present analysis. Ninety-one patients (53.5%) had loneliness, and the mean TMIG-IC score was 11.3. The unadjusted and adjusted partial regression coefficients of loneliness on higher-level functions were -1.61 (95% confidence interval [CI], -2.31 to -0.91; < 0.001) and -0.88 (95% CI, -1.52 to -0.23; P=0.008), respectively. CONCLUSION: Loneliness in elderly diabetic patients was found to be associated with lower higher-level functions. It is important to remind patients with diabetes mellitus who have loneliness about the risk of a decline in their higher-level functions.

Loss of PRMT1 in the central nervous system (CNS) induces reactive astrocytes and microglia during postnatal brain development.

PRMT1, a major arginine methyltransferase, plays critical roles in transcription, DNA damage response, and cell proliferation. Although we have previously discovered the crucial roles of PRMT1 for oligodendrocyte lineage progression in the central nervous system of neural stem cell-specific PRMT1 conditional knockout (PRMT1-CKO) mice, the context of other glial cell states that may cause the hypomyelination phenotype in PRMT1-CKO mice has not been explored so far. Here, we performed RNA-seq of the neonatal cortices of PRMT1-CKO mice to reveal overall gene expression changes and show the up-regulation of inflammatory signaling which is generally mediated by astrocytes and microglia in advance of the myelination defects. In particular, qRT-PCR analyses revealed Interleukin-6 (Il-6), a major central nervous system cytokine, was dramatically increased in the PRMT1-CKO brains. The gene expression changes led to augmentation of glial fibrillary acidic protein and Vimentin protein levels in PRMT1-CKO mice, showing severe reactive astrogliosis after birth. We further show that IBA1-positive and CD68-positive activated microglia were increased in PRMT1-CKO mice, in spite of intact Prmt1 gene expression in purified microglia from the mutant mice. Our results indicate that PRMT1 loss in the neural stem cell lineage causes disruptive changes in all glial types perturbing postnatal brain development and myelination.

Impact of diagnosis-to-ablation time on non-pulmonary vein triggers and ablation outcomes in persistent atrial fibrillation.

INTRODUCTION: Non-pulmonary vein (PV) triggers are a major cause of atrial tachyarrhythmia (ATA) recurrence after catheter ablation. However, the effect of the diagnosis-to-ablation time (DAT) on non-PV triggers in persistent atrial fibrillation is unknown. METHODS AND RESULTS: This observational study evaluated 502 consecutive persistent AF patients who underwent initial ablation. We compared 408 patients whose DAT was <3 years with 94 patients whose DAT was ≥3 years. Following PV and posterior wall isolation, 193 non-PV triggers, including 50 AFs, 30 atrial tachycardias (ATs), and 113 repetitive atrial premature beats, were elicited and ablated in 137 (27%) patients. Specifically, 80 non-PV AF/AT triggers were provoked in 64 (13%) patients, being identified more frequently in the DAT ≥ 3 years group than in the DAT < 3 years group (20% vs. 11%, p = .025) especially with a higher prevalence of coronary sinus/inferior left atrial triggers. During a median follow-up of 770 days, the ATA recurrence-free rate was higher in the DAT < 3 years group than the DAT ≥ 3 years group (79% vs. 53% at 2 years, p < .001). In a multivariate analysis, female sex (odds ratio: 2.70, p = .002) and a longer DAT (odds ratio: 1.13/year, p = .008) were predictors of non-PV AF/AT triggers, and a longer DAT (hazard ratio: 1.12/year, p < .001) and non-PV AT/AF triggers (hazard ratio: 1.79, p = .009) were associated with ATA recurrence. CONCLUSION: Early ablation after the first diagnosis of persistent AF may reduce emerging non-PV AF/AT triggers and ATA recurrence.