Single-step generation of homozygous knockout/knock-in individuals in an extremotolerant parthenogenetic tardigrade using DIPA-CRISPR.

Tardigrades are small aquatic invertebrates known for their remarkable tolerance to diverse extreme stresses. To elucidate the in vivo mechanisms underlying this extraordinary resilience, methods for genetically manipulating tardigrades have long been desired. Despite our prior success in somatic cell gene editing by microinjecting Cas9 ribonucleoproteins (RNPs) into the body cavity of tardigrades, the generation of gene-edited individuals remained elusive. In this study, employing an extremotolerant parthenogenetic tardigrade species, Ramazzottius varieornatus, we established conditions that led to the generation of gene-edited tardigrade individuals. Drawing inspiration from the direct parental CRISPR (DIPA-CRISPR) technique employed in several insects, we simply injected a concentrated Cas9 RNP solution into the body cavity of parental females shortly before their initial oviposition. This approach yielded gene-edited G0 progeny. Notably, only a single allele was predominantly detected at the target locus for each G0 individual, indicative of homozygous mutations. By co-injecting single-stranded oligodeoxynucleotides (ssODNs) with Cas9 RNPs, we achieved the generation of homozygously knocked-in G0 progeny, and these edited alleles were inherited by G1/G2 progeny. This is the first example of heritable gene editing in the entire phylum of Tardigrada. This establishment of a straightforward method for generating homozygous knockout/knock-in individuals not only facilitates in vivo analyses of the molecular mechanisms underpinning extreme tolerance, but also opens up avenues for exploring various topics, including Evo-Devo, in tardigrades.

Age-dependent analysis dissects the stepwise control of auxin-mediated lateral root development in rice.

As root elongation rates are different among each individual root, the distance from the root apices does not always reflect the age of root cells. Thus, methods for correcting variations in elongation rates are needed to accurately evaluate the root developmental process. Here, we show that modeling-based age-dependent analysis is effective for dissecting stepwise lateral root (LR) development in rice (Oryza sativa). First, we measured the increases in LR and LR primordium (LRP) numbers, diameters, and lengths in wild type and an auxin-signaling-defective mutant, which has a faster main (crown) root elongation rate caused by the mutation in the gene encoding AUXIN/INDOLE-3-ACETIC ACID protein 13 (IAA13). The longitudinal patterns of these parameters were fitted by the appropriate models and the age-dependent patterns were identified using the root elongation rates. As a result, we found that LR and LRP numbers and lengths were reduced in iaa13. We also found that the duration of the increases in LR and LRP diameters were prolonged in iaa13. Subsequent age-dependent comparisons with gene expression patterns suggest that AUXIN RESPONSE FACTOR11 (ARF11), the homolog of MONOPTEROS (MP)/ARF5 in Arabidopsis (Arabidopsis thaliana), is involved in the initiation and growth of LR(P). Indeed, the arf11 mutant showed a reduction of LR and LRP numbers and lengths. Our results also suggest that PINOID-dependent rootward-to-shootward shift of auxin flux contributes to the increase in LR and LRP diameters. Together, we propose that modeling-based age-dependent analysis is useful for root developmental studies by enabling accurate evaluation of root traits' expression.

Stress-dependent cell stiffening by tardigrade tolerance proteins that reversibly form a filamentous network and gel.

Tardigrades are able to tolerate almost complete dehydration by entering a reversible ametabolic state called anhydrobiosis and resume their animation upon rehydration. Dehydrated tardigrades are exceptionally stable and withstand various physical extremes. Although trehalose and late embryogenesis abundant (LEA) proteins have been extensively studied as potent protectants against dehydration in other anhydrobiotic organisms, tardigrades produce high amounts of tardigrade-unique protective proteins. Cytoplasmic-abundant heat-soluble (CAHS) proteins are uniquely invented in the lineage of eutardigrades, a major class of the phylum Tardigrada and are essential for their anhydrobiotic survival. However, the precise mechanisms of their action in this protective role are not fully understood. In the present study, we first postulated the presence of tolerance proteins that form protective condensates via phase separation in a stress-dependent manner and searched for tardigrade proteins that reversibly form condensates upon dehydration-like stress. Through a comprehensive search using a desolvating agent, trifluoroethanol (TFE), we identified 336 proteins, collectively dubbed "TFE-Dependent ReversiblY condensing Proteins (T-DRYPs)." Unexpectedly, we rediscovered CAHS proteins as highly enriched in T-DRYPs, 3 of which were major components of T-DRYPs. We revealed that these CAHS proteins reversibly polymerize into many cytoskeleton-like filaments depending on hyperosmotic stress in cultured cells and undergo reversible gel-transition in vitro. Furthermore, CAHS proteins increased cell stiffness in a hyperosmotic stress-dependent manner and counteract the cell shrinkage caused by osmotic pressure, and even improved the survival against hyperosmotic stress. The conserved putative helical C-terminal region is necessary and sufficient for filament formation by CAHS proteins, and mutations disrupting the secondary structure of this region impaired both the filament formation and the gel transition. On the basis of these results, we propose that CAHS proteins are novel cytoskeleton-like proteins that form filamentous networks and undergo gel-transition in a stress-dependent manner to provide on-demand physical stabilization of cell integrity against deformative forces during dehydration and could contribute to the exceptional physical stability in a dehydrated state.

Cellular stress induces non-canonical activation of the receptor tyrosine kinase EphA2 through the p38-MK2-RSK signaling pathway.

The receptor tyrosine kinase ephrin type-A receptor 2 (EphA2) is overexpressed in malignant tumors. We previously reported that non-canonical EphA2 phosphorylation at Ser-897 was catalyzed by p90 ribosomal S6 kinase (RSK) via the MEK-ERK pathway in ligand- and tyrosine kinase-independent manners. Non-canonical EphA2 activation plays a key role in tumor progression; however, its activation mechanism remains unclear. In the present study, we focused on cellular stress signaling as a novel inducer of non-canonical EphA2 activation. p38, instead of ERK in the case of epidermal growth factor signaling, activated RSK-EphA2 under cellular stress conditions, including anisomycin, cisplatin, and high osmotic stress. Notably, p38 activated the RSK-EphA2 axis via downstream MAPK-activated protein kinase 2 (MK2). Furthermore, MK2 directly phosphorylated both RSK1 Ser-380 and RSK2 Ser-386, critical residues for the activation of their N-terminal kinases, which is consistent with the result showing that the C-terminal kinase domain of RSK1 was dispensable for MK2-mediated EphA2 phosphorylation. Moreover, the p38-MK2-RSK-EphA2 axis promoted glioblastoma cell migration induced by temozolomide, a chemotherapeutic agent for the treatment of glioblastoma patients. Collectively, the present results reveal a novel molecular mechanism for non-canonical EphA2 activation under stress conditions in the tumor microenvironment.

Renin-Angiotensin-Aldosterone System Inhibitors Prevent the Onset of Oxaliplatin-Induced Peripheral Neuropathy: A Retrospective Multicenter Study and in Vitro Evaluation.

Oxaliplatin (OXA) is used in chemotherapy for various cancer types and is associated with acute and chronic neurotoxicity. However, a preventive strategy for OXA-induced peripheral neuropathy (OIPN) and its underlying mechanism remain unclear. We examined the effects of renin-angiotensin-aldosterone system inhibitors (RAASIs) on OIPN by performing a retrospective multicenter study and an in vitro assay. We retrospectively evaluated electronic medical records of 976 patients who underwent one or more courses of OXA-containing regimens at Ehime, Okayama, and Tokushima University Hospitals. The primary endpoint was the incidence of OIPN during or after OXA administration. The effects of RAASIs and OXA on the neurite length in PC12 cells were determined. The combined administration of an OXA-containing regimen and RAASI significantly inhibited the cumulative incidence grade-2 or higher OIPN (log-rank test; p = 0.0001). RAASIs markedly suppressed the development of both acute and chronic OIPN (multivariate analysis; p = 0.017 and p = 0.011). In an in vitro assay, 10 µM OXA suppressed the neurite length; treatment with 1 µM aliskiren, spironolactone, 10 µM candesartan, and enalapril significantly restored neurite length to the control level. Moreover, 1 µM SCH772984 (a selective inhibitor of extracellular signal-regulated kinase, ERK1/2) and 500 µM SQ22536 (a cell-permeable adenylate cyclase (AC) inhibitor) markedly abolished neurite-extending effects of candesartan and enalapril. These results indicate that RAASIs possess preventive or therapeutic effects in acute and chronic OIPN, candesartan and enalapril may increase in the activity of ERK1/2 and AC in PC12 cells.

Fine control of aerenchyma and lateral root development through AUX/IAA- and ARF-dependent auxin signaling.

Lateral roots (LRs) are derived from a parental root and contribute to water and nutrient uptake from the soil. Auxin/indole-3-acetic acid protein (AUX/IAA; IAA) and auxin response factor (ARF)-mediated signaling are essential for LR formation. Lysigenous aerenchyma, a gas space created by cortical cell death, aids internal oxygen transport within plants. Rice (Oryza sativa) forms lysigenous aerenchyma constitutively under aerobic conditions and increases its formation under oxygen-deficient conditions; however, the molecular mechanisms regulating constitutive aerenchyma (CA) formation remain unclear. LR number is reduced by the dominant-negative effect of a mutated AUX/IAA protein in the iaa13 mutant. We found that CA formation is also reduced in iaa13 We have identified ARF19 as an interactor of IAA13 and identified a lateral organ boundary domain (LBD)-containing protein (LBD1-8) as a target of ARF19. IAA13, ARF19, and LBD1-8 were highly expressed in the cortex and LR primordia, suggesting that these genes function in the initiation of CA and LR formation. Restoration of LBD1-8 expression recovered aerenchyma formation and partly recovered LR formation in the iaa13 background, in which LBD1-8 expression was reduced. An auxin transport inhibitor suppressed CA and LR formation, and a natural auxin stimulated CA formation in the presence of the auxin transport inhibitor. Our findings suggest that CA and LR formation are both regulated through AUX/IAA- and ARF-dependent auxin signaling. The initiation of CA formation lagged that of LR formation, which indicates that the formation of CA and LR are regulated differently by auxin signaling during root development in rice.

Suitable Dose of Long-Term Tolvaptan to Reduce Heart Failure Rehospitalizations.

The short-term effectiveness of tolvaptan (TLV) for heart failure (HF) has been established, but the long-term effects are controversial. We investigated HF patients who could not discontinue both loop diuretics and TLV at discharge from AURORA (Acute Heart Failure Registry in Osaka Rosai Hospital). We compared the following factors at discharge between the RH group, consisting of patients with rehospitalizations due to worsening HF within 1 year after discharge (RH group), and non-RH group: age, gender, blood pressure, history of HF admission, electrocardiogram and echocardiographic parameters, atherosclerotic risk factors, laboratory data, and medications. Furthermore, we compared the effects of long-term low-dose TLV (≤ 7.5 mg/day) and high-dose TLV on HF rehospitalizations. The RH group consisted of 81 patients (58.7%). A multivariate analysis revealed that a history of HF admission and the TLV dose were independently and significantly associated with 1-year HF rehospitalizations. A receiver operating characteristic curve revealed that 7.5 mg of TLV was a suitable cutoff value for 1-year HF rehospitalizations. The Kaplan-Meier curves demonstrated that the HF rehospitalization free ratio was significantly higher in the low-dose TLV group (≤ 7.5 mg/day) than in high-dose TLV group over 1 year.In conclusion, the TLV dose, in addition to a history of HF admission, was associated with 1-year HF rehospitalizations in diuretic-dependent HF patients. In these patients, long-term low-dose TLV (≤ 7.5 mg/day) may be favorable for reducing HF rehospitalizations.

A modified GnRH antagonist method in combination with letrozole, cabergoline, and GnRH antagonist for PCOS: Safe and effective ovarian stimulation to treat PCOS and prevent OHSS.

PURPOSE: To analyze the therapeutic efficacy of a modified controlled ovarian stimulation (COS) protocol for polycystic ovary syndrome (PCOS) that does not cause ovarian hyperstimulation syndrome (OHSS) while maintaining oocyte quality. METHOD: This study is a retrospective cohort study of reproductive medicine at St. Mother Clinic. We analyzed ART clinical outcomes, embryonic development, and hormone levels in 175 PCOS patients treated with four COS (GnRH agonist based long protocol, Group A; GnRH antagonist protocol with HCG trigger, Group B; GnRH antagonist protocol with GnRH agonist trigger, Group C, and the modified COS group) between 2010 and 2021. RESULTS: Of 175 patients with PCOS, 45 and 130 patients underwent 47 and 136 oocyte retrieval cycles, 75 and 250 embryo transfer cycles with the modified COS, and with conventional methods, respectively. The cumulative pregnancy rate at one trial was a significantly higher result than in Group A and higher than in Groups B and C (cumulative pregnancy rate at one trial of Group A, B, C, and modified COS: 40.0%, 54.5%, 56.3%, and 72.3%, respectively). With this method, not clinically problematic OHSS and higher clinical outcomes than in conventional methods were observed. CONCLUSION: This modified COS can significantly improve clinical outcomes and eliminate OHSS.

Electrophysiological identification of superior vena cava: Novel insight into slow conduction or conduction block.

INTRODUCTION: It has not been clarified how to identify the electrophysiological junction between right atrium (RA) and superior vena cava (SVC). The aim of this study was to identify the electrophysiological RA-SVC junction according to slow conduction or conduction bock and to examine the electrophysiological SVC isolation procedure. METHODS: Seventy-three consecutive atrial fibrillation patients who underwent SVC mapping using a CARTO 3 system were enrolled in this study. Slow conduction or conduction block between the RA and SVC was identified by adjusting the lower threshold criteria of the early meets late function and was described as a white line. The SVC isolation was performed along the white line and with pacing maneuvers to confirm direct SVC capture. RESULTS: Activation mapping (1296 ± 631 points) was obtained in 66 patients (90%) in 4.6 ± 1.8 min. Slow conduction or conduction block was observed in all patients. The threshold for detecting slow conduction or conduction block was 24 ± 8 ms. The location of the electrophysiological RA-SVC junction was higher in the anterior portion (anterior-septal, anterior, and anterior-lateral) than in the posterior portion (posterior-septal, posterior, and posterior-lateral) (-2.3 ± 6.2 mm vs. 7.1 ± 6.3 mm, p < .001). The SVC isolation at the electrophysiological RA-SVC junction was successful in all patients without any injury to the sinus node function. Asymptomatic phrenic nerve injury was observed in three patients (4.5%). CONCLUSION: In all patients, the electrophysiological RA-SVC junction determined by slow conduction or conduction block was identified and the electrophysiological SVC isolation was performed successfully and safely.

Effect of the Balloon Size on Lesion Formation During Visually Guided Laser Balloon Ablation in an In Vitro Model.

BACKGROUND: It is unclear whether balloon size can influence lesion formation. The aim of this study was to evaluate the impact of balloon size on lesion formation during laser balloon procedures in an in vitro model.Methods and Results:Laser energy was applied to chicken muscle using a first generation laser balloon. Laser ablation was performed with 2 different balloon sizes (18 mm and 32 mm) using 2 different power settings (12 W/20 s and 8.5 W/20 s) on the chicken muscle. The lesion characteristics, including maximum lesion depth, maximum lesion diameter, surface diameter and depth at maximum diameter, were compared between the 18-mm and 32-mm balloon groups at 12 W/20 s and 8.5 W/20 s, respectively. We created 40 lesions using laser energy at 12 W/20 s and 80 lesions at 8.5 W/20 s. At both power settings, the maximum lesion depth and the depth at the maximum diameter were larger in the 18-mm than in the 32-mm balloon group. At both power settings, the maximum lesion diameter and the surface diameter were smaller in the 18-mm than in the 32-mm balloon group. CONCLUSIONS: The balloon size could affect the lesion formation during laser balloon ablation. The lesion with the larger balloon size was wider and shallower than the lesion with the smaller balloon size.

Predictors of improvement of left ventricular systolic function after catheter ablation of persistent atrial fibrillation in patients with heart failure with reduced ejection fraction.

Although several studies have suggested that catheter ablation (CA) of atrial fibrillation (AF) can improve left ventricular ejection fraction (LVEF) in patients with heart failure with reduced ejection fraction (HFrEF), the predictor of improvement of LVEF is unclear. A total of 401 patients with persistent AF underwent an initial CA between September 2014 and October 2019 in our hospital. Among them, we analyzed consecutive patients with moderately or severely reduced LVEF (< 50%) measured by transthoracic echocardiography (TTE) within 2 months before CA and underwent follow-up TTE during sinus rhythm at 6 months or more after CA. These patients were categorized into two groups: improve group (I group) with the absolute improvement of LVEF ≥ 10% at follow-up TTE, and non- improve group (NI group) with the absolute improvement of LVEF < 10% at follow-up TTE. We compared patient characteristics, ablation procedures, and clinical outcomes between the two groups. 81 patients were analyzed, and I group consisted of 48 patients (59%). In the univariate analysis, absence of ischemic cardiomyopathy, left ventricular end-diastolic diameter (LVEDD), and absence of recurrence of AF between 3 and 6 months after CA were associated with improvement of LVEF. A receiver operating characteristics analysis determined the suitable cut-off value for LVEDD was 53 mm (sensitivity: 62.2%, specificity: 86.2%, area under curve: 0.762). A multivariate analysis showed that LVEDD < 53 mm was independently associated with improvement of LVEF (odds ratio 2.58, 95% confidence interval 1.29-6.12; P = 0.021). In conclusion, LVEDD < 53 mm might be an independent predictor of improvement of LVEF after CA of persistent AF in HFrEF patients.

Impact of urinary albumin excretion on the onset of adverse reactions to vancomycin hydrochloride.

OBJECTIVE: Vancomycin dose needs to be reduced for decreased kidney function; however, impact of urinary albumin excretion (UAE) on serum vancomycin level is unknown. In this study, we examined the factors associated with the onset of renal impairment induced by vancomycin by focusing on UAE. MATERIALS AND METHODS: The study included 52 patients who received vancomycin for methicillin-resistant Staphylococcus aureus at Ehime University Hospital between April 2010 and March 2015. To determine the presence of renal impairment, patients whose common terminology criteria for adverse-events (CTCAE) grade worsened by 1 or more comprised the renal impairment group, and multivariate logistic regression analysis was performed. RESULTS: 13 patients (25%) had renal impairment as indicated by a CTCAE grade change by 1 or more. The results of multivariate logistic regression analysis only of UAE (OR = 18.03; 95% CI = 1.97 - 164.89) was identified as a significant factor. CONCLUSION: We investigated the factors associated with the onset of renal impairment induced by vancomycin and identified UAE as a potential risk factor.

Application of Quantitative NMR Spectroscopy to the Quality Evaluation of Diclofenac Gargles as Hospital Preparations.

Hospital preparations are frequently prepared in Japanese hospitals when ready-made formulations to meet patients' needs are unavailable. Although the quality of hospital preparations have to be ensured for efficacy and safety, such quality evaluation tends to be insufficient mainly due to lack of manpower and experimental environments in hospitals. In this paper, we investigated the applicability of quantitative (q)NMR spectroscopy to the quality control of diclofenac gargles as examples of hospital preparations, as it has various merits for the quantitative analysis of mixtures in solutions. Diclofenac gargles are composed of diclofenac, tranexamic acid, and lidocaine, and are used for the pain relief of stomatitis induced by cancer chemotherapy. Aliquots of the gargles, which were prepared five times, were mixed with dimethylsulfone as an internal standard, followed by qNMR measurements. Water signal suppression was achieved using a pulse program, water suppression enhanced through T1 effects, because the pulse program was superior to other ones such as presaturation and one-dimensional nuclear Overhauser effect spectroscopy in terms of quantitativeness. Concentrations of the three medicinal ingredients were simultaneously determined based on the signals selected by considering the spectral separation and the quantitativeness. Consequently, the gargles were found to be prepared with constant quality, and were stable at room temperature for at least four weeks. qNMR is considered to be potentially useful for the quality control of various hospital preparations because of minimal sample pretreatments, lack of need of calibration curves, and its comprehensive detection abilities.

Culture shapes emotion perception from faces and voices: changes over development.

The perception of multisensory emotion cues is affected by culture. For example, East Asians rely more on vocal, as compared to facial, affective cues compared to Westerners. However, it is unknown whether these cultural differences exist in childhood, and if not, which processing style is exhibited in children. The present study tested East Asian and Western children, as well as adults from both cultural backgrounds, to probe cross-cultural similarities and differences at different ages, and to establish the weighting of each modality at different ages. Participants were simultaneously shown a face and a voice expressing either congruent or incongruent emotions, and were asked to judge whether the person was happy or angry. Replicating previous research, East Asian adults relied more on vocal cues than did Western adults. Young children from both cultural groups, however, behaved like Western adults, relying primarily on visual information. The proportion of responses based on vocal cues increased with age in East Asian, but not Western, participants. These results suggest that culture is an important factor in developmental changes in the perception of facial and vocal affective information.

Underweight Is Associated with Poor Prognosis in Heart Failure with Preserved Ejection Fraction.

The obesity paradox states higher body mass index (BMI) is associated with better outcomes than normal weight in patients with heart failure with preserved ejection fraction (HFpEF). However, underweight was defined by BMI < 18.5 kg/m2, and results have been inconclusive, in part due to small number of participants. The number of underweight patients with HFpEF is higher in Asian than in Western countries. In this study, we aim to determine the prognostic impact of underweight in patients with HFpEF in Asian population.We enrolled 846 consecutive patients from the PURSUIT-HFpEF registry. We then divided them into three groups by BMI, namely, underweight (BMI < 18.5 kg/m2), normal weight (18.5 ≤ BMI < 23), and overweight (23 ≤ BMI). The underweight group consisted of 187 patients (22%). Over a mean follow-up of 407 days, 105 deaths were reported as all-cause mortality. On multivariable Cox analysis, the underweight group was determined to be significantly associated with higher risk of all-cause mortality than the normal and overweight groups (Hazard ratios [HR]: 2.33; 95% confidence intervals [CI]: 1.45-3.75, P < 0.001; HR: 3.54; 95% CI: 1.99-6.29, P < 0.001, respectively), after adjustment for age, sex, vital signs, and comorbidities.Underweight is a useful predictor of poor prognosis in patients with HFpEF in Asian population.

Statins can suppress DC-mediated Th2 responses through the repression of OX40-ligand and CCL17 expression.

DCs and epithelial cell-derived thymic stromal lymphopoietin (TSLP) have pivotal roles in allergic inflammation. TSLP stimulates myeloid DCs to express OX40-ligand (OX40L) and CCL17, which trigger and maintain Th2 cell responses. We have previously shown that statins, which are HMG-CoA reductase inhibitors, have the ability to suppress type I IFN production by plasmacytoid DCs. Here, we extended our previous work to examine the immunomodulatory effect of statins on allergic responses, particularly the TSLP-dependent Th2 pathway induced by myeloid DCs. We found that treatment of TSLP-stimulated DCs with either pitavastatin or simvastatin suppressed both the DC-mediated inflammatory Th2 cell differentiation and CRTH2+ CD4+ memory Th2 cell expansion and also repressed the expressions of OX40L and CCL17 by DCs. These inhibitory effects of statins were mimicked by treatment with either a geranylgeranyl-transferase inhibitor or Rho-kinase inhibitor and were counteracted by the addition of mevalonate, suggesting that statins induce geranylgeranylated Rho inactivation through a mevalonate-dependent pathway. We also found that statins inhibited the expressions of phosphorylated STA6 and NF-κB-p50 in TSLP-stimulated DCs. This study identified a specific ability of statins to control DC-mediated Th2 responses, suggesting their therapeutic potential for treating allergic diseases.

Comparison of myocardial injury and inflammation after pulmonary vein isolation for paroxysmal atrial fibrillation between radiofrequency catheter ablation and cryoballoon ablation.

BACKGROUNDS: Several studies have shown the serum high sensitive cardiac troponin I (hs-TnI) a biomarker of myocardium injury, and C-reactive protein (CRP), a biomarker of inflammation, are associated with worse cardiovascular outcomes. We evaluated the relationship between the hs-TnI level in patients with paroxysmal atrial fibrillation (PAF) after pulmonary vein isolation (PVI) and atrial fibrillation (AF) recurrence. METHODS AND RESULTS: We enrolled 263 consecutive PAF patients who underwent PVI from May 2017 to April 2018. We investigated the difference in the relationship between the myocardial injury marker (serum hs-TnI), inflammatory marker (CRP, white blood cell) at 36 to 48 hours after the PVI, and early or late recurrence of AF (ERAF; <3 months and LRAF; from 3 months to 1 year) between the radiofrequency ablation group (R group) and cryoballoon ablation group (C group). The R group consisted of 147 patients and the C groups consisted of 116 patients. The serum hs-TnI level in R group was significantly lower than in the C group (2.33 vs 5.08 ng/mL; P < .001), while the CRP was significantly higher in the R group than C group (2.02 vs 1.10 mg/dL; P < .001). The incidences of an ERAF/LRAF were similar between the two groups. CONCLUSION: Cryoballoon ablation may cause more myocardial injury than radiofrequency catheter ablation, on the contrary, radiofrequency catheter ablation, may cause more inflammation than cryoballoon ablation. However, these phenomena may not affect the recurrence of AF after the PVI in patient with PAF.

Sulfonamide antibiotics inhibit RNAi by binding to human Argonaute protein 2 PAZ.

We found that sulfisomidine, a sulfonamide antibiotic, potently binds to the Piwi/Argonaute/Zwille (PAZ) domain of human Argonaute protein 2 and inhibits RNA interference (RNAi). To elucidate the effect on RNAi of strong affinity of the 3'-ends in small interfering RNA (siRNA) to the PAZ domain, chemically modified siRNAs bearing sulfisomidine at the 3'-end were synthesized.

Clinical experience of uninterrupted oral anticoagulants during atrial fibrillation ablation in elderly patients: a comparison with non-elderly patients.

It is unclear whether uninterrupted oral anticoagulants (OACs) are acceptable in elderly patients as compared to non-elderly patients. This study aimed to assess the clinical impact of an uninterrupted OAC strategy during atrial fibrillation (AF) ablation in elderly patients. We enrolled 439 consecutive patients who underwent AF ablation with an uninterrupted OAC strategy at our institute. The incidence of bleeding and thromboembolic complications during the AF ablation or within 4 weeks after and the anticoagulation status during the AF ablation were compared between the elderly (age ≧ 75, n = 144) and non-elderly groups (n = 295). There were 22 bleeding complications in the elderly group and 31 in the non-elderly group (15% vs. 11%, p = 0.162) and 3 major bleeding complications in the elderly group and 2 in the non-elderly group (2% vs. 0.7%, p = 0.336). Thromboembolic events were observed in 1 patient in the elderly group and 2 in the non-elderly group (0.7% vs. 0.7%, p = 1.000). The ACT at the end of the procedure was longer in the elderly group than in the non-elderly group (350 s vs. 341 s, p = 0.007) and the proportion of a prolonged ACT of > 400 s (27% vs. 18%, p = 0.046) was more frequent in the elderly group than non-elderly group. A propensity score matched population excluding the age and body weight, revealed that the anticoagulation status during AF ablation was comparable between the two groups. Thus, in the patients undergoing AF ablation with uninterrupted OAC strategy, bleeding complications in elderly patients were similar to those in non-elderly patients. The anticoagulation status during the procedure in elderly patients was more prolonged than that in non-elderly patients.