SpCas9-HF1 enhances accuracy of cell cycle-dependent genome editing by increasing HDR efficiency, and by reducing off-target effects and indel rates.

In genome editing, it is important to avoid off-target mutations so as to reduce unexpected side effects, especially for therapeutic applications. Recently, several high-fidelity versions of SpCas9 have been developed to reduce off-target mutations. In addition to reducing off-target effects, highly efficient intended target gene correction is also essential to rescue protein functions that have been disrupted by single nucleotide polymorphisms. Homology-directed repair (HDR) corrects genes precisely using a DNA template. Our recent development of cell cycle-dependent genome editing has shown that regulation of Cas9 activation with an anti-CRISPR-Cdt1 fusion protein increases HDR efficiency and reduces off-target effects. In this study, to apply high-fidelity SpCas9 variants to cell cycle-dependent genome editing, we evaluated anti-CRISPR inhibition of high-fidelity SpCas9s. In addition, HDR efficiency of high-fidelity SpCas9s was addressed, identifying eSpCas9, SpCas9-HF1, and LZ3 Cas9 as promising candidates. Although eSpCas9 and LZ3 Cas9 showed decreased HDR efficiency in cell cycle-dependent genome editing, SpCas9-HF1 successfully achieved increased HDR efficiency and few off-target effects when co-expressed with an AcrIIA4-Cdt1 fusion.

Combination of robot-assisted laparoscopic pyeloplasty for lower moiety ureteropelvic junction obstruction in a partial duplex system and percutaneous endoscopic surgery for renal calculi reusing the port for robotic pyeloplasty.

Introduction: Ureteropelvic junction obstruction is often associated with renal calculus formation. However, there is no report of using robot-assisted laparoscopic pyeloplasty combined with percutaneous endoscopic surgery for ureteropelvic junction obstruction and renal calculi in a partial duplex system. Case presentation: A 19-year-old female patient with lower moiety ureteropelvic junction obstruction and renal calculi in a partial duplex system was referred to our hospital because of left lumbar pain, left acute pyelonephritis, and an increase in left renal calculi during follow-up at the referral hospital. To prevent the complication of percutaneous nephrolithotripsy following pyeloplasty, robot-assisted laparoscopic pyeloplasty combined with percutaneous endoscopic surgery was performed. Two years after surgery, the patient reported no left lumbar pain. Conclusion: The combination of robot-assisted laparoscopic pyeloplasty and percutaneous endoscopic surgery can be proposed as a safe and less-invasive treatment option for ureteropelvic junction obstruction and renal calculi in a partial duplex system.

Pseudo dilated cardiomyopathy: Dilated cardiomyopathy-like changes due to a combination of stuck mechanical mitral valve and coronary microvascular dysfunction-An autopsy case.

Thrombus formation in the microvessels and endocardium was suggestive of endothelial cell damage, myocardial ischemia, and a decreased coronary flow reserve. Sustained pulmonary hypertension due to thrombosis worsened the biventricular dysfunction.

Gap between the prevalence of and consultation rate for lumbopelvic pain in postnatal Japanese women.

BACKGROUND: We aimed to assess the prevalence of lumbopelvic pain (LPP), including low back pain (LBP) and/or pelvic girdle pain (PGP), consultation rate, and desire for treatment of postnatal Japanese women. METHODS: We performed a cross-sectional study of 98 postnatal Japanese women within 1 year of childbirth. We retrospectively evaluated the prevalence of LBP before and during pregnancy and the prevalence of LPP after childbirth using a self-administered questionnaire. We used the distribution of pain to differentiate LBP and/or PGP after childbirth and its intensity was assessed using a visual analogue scale. The effects of LPP on daily life were assessed using the Oswestry disability index (ODI). In addition, we assessed the consultation rate for LPP and the desire to consult a healthcare professional, using self-administered questionnaires. RESULTS: The prevalence of LPP after childbirth was 66%. Women with a history of LBP before and during pregnancy were more likely to have LPP following childbirth (both P < 0.001). The consultation rate among the postnatal women with LPP was 9%, but 56% of the women wished to consult a healthcare professional. The ODI score was significantly higher in postnatal women who wanted to consult a healthcare professional than in those who did not (P < 0.01). CONCLUSIONS: Two thirds of the postnatal Japanese women who participated in this study had LPP, but the consultation rate was low. However, more than half of these women had the desire to consult a healthcare professional.

Acute myocarditis secondary to Campylobacter jejuni enteritis: A case report.

Acute myocarditis is a rare complication of Campylobacter jejuni enteritis. Herein, we report the case of a 20-year-old man who presented with chest pain that developed three days after the onset of enteritis. Electrocardiogram, echocardiogram, and cardiac enzyme levels suggested myocarditis. Cardiac magnetic resonance imaging revealed a late gadolinium enhancement in the inferior wall. Degeneration and necrosis of myocardial cells and lymphocyte-dominant inflammatory cell infiltration were found in the tissue obtained by endomyocardial biopsy. Acute myocarditis associated with C. jejuni enteritis was confirmed by these findings and C. jejuni detected in the stool culture. The symptoms of enteritis and myocarditis remitted 10 days after the onset. The left ventricular ejection fraction was improved from 40 % to 57 %.In previous cases, endomyocardial biopsy has not been performed because of mild myocarditis. The lack of pathological reports makes the mechanism of myocarditis associated with C. jejuni enteritis unknown. We report a case of myocarditis associated with C. jejuni enteritis, which was diagnosed using cardiac magnetic resonance imaging and endomyocardial biopsy. Learning objective: Acute myocarditis is a rare but important complication of Campylobacter jejuni enteritis. Cardiac magnetic resonance imaging is useful for diagnosis. Most cases of myocarditis associated with C. jejuni enteritis were mild and remitted without specific treatment. In the present case, endomyocardial biopsy was performed and CD4-positive lymphocytes were predominantly detected in the myocardial tissue.

The association between early rehabilitation and ambulatory ability at discharge in patients with hip fractures at acute-phase rehabilitation wards: a survey of the Japan Association of Rehabilitation Database.

This study aimed to examine the effectiveness of early rehabilitation in patients with femoral neck fractures admitted to acute care settings in Japan using the data registered with the Japan Association of Rehabilitation Databases (JARD). We included data for 401 patients (out of 3088 patients) aged ≥ 65 years (85 males, 316 females) from nine hospitals who sustained a femoral neck fracture between July 2005 and September 2015. Using the number of days until surgery or the number of days until the start of rehabilitation or both as the explanatory variables, and the indoor mobility at discharge as the outcome variable, we calculated the adjusted rate ratio (ARR) and 95% confidence interval (CI) using Poisson regression analysis (age, sex, cognitive impairment, concurrent symptoms, and previous history of fracture adjusted as covariates). The ARR for independent walking at the discharge of the early-rehabilitation group (starting rehabilitation within two days after the injury) was significantly higher (ARR: 2.01, 95% CI: 1.34-3.02) than that of the non-early rehabilitation group. These results suggest that early acute-phase rehabilitation after a femoral neck fracture in older patients allows for better ambulatory ability at discharge, regardless of the time to surgery.

Lung regeneration with rat fetal lung implantation and promotion of alveolar stem cell differentiation by corticosteroids.

Introduction: The lung is a difficult organ to regenerate, and the development of functional lungs has still not been achieved. In this study, we investigated lung regeneration using a rat fetal lung tissue-implanted model. This study aimed to evaluate the functioning of the implanted fetal lung tissue and investigate the graft differentiation and maturation mechanism, focusing on alveolar stem cells. Methods: Fetal lung tissue fragments were obtained from Lewis rats on day 17 and implanted into adult lungs. Animals were divided into the following three groups: group 1, injection into the adult left lung parenchyma; group 2, injection with post-caval lobectomy; and group 3, injection with post-caval lobectomy and corticosteroid administration. Computed tomography was performed on weeks 1, 2, 4, and 8. The presence of alveolar pore, CD31 expression, and bipotential progenitor cell (podoplanin+/surfactant protein C+) localization were histologically evaluated. MiRNA expression was comprehensively compared among the three groups. Results: The grafts comprised type I and type II alveolar cells connected to the recipient lungs with alveolar pores and capillary networks in the interstitial tissue. The alveolar space was the largest and the computed tomography value was the lowest in the grafts of the corticosteroid-administered group. The number of bipotential progenitor cells was the lowest in the corticosteroid administration group on day 7. Moreover, microRNA-487-3p, 374-5p, and 20b-5p expression was changed by more than 2-fold between the post-caval lobectomy and corticosteroid administration groups. Conclusions: Implanted fetal lung tissues established airway and capillary communication with the recipient lungs, and corticosteroids accelerated their maturation by promoting the differentiation of progenitor cells. The study findings provide new insights into lung regeneration research.

The history of genome editing: advances from the interface of chemistry & biology.

Genome editing had a long history before the appearance of CRISPR. Although a decade has passed since the initial use of CRISPR with mammalian cells, the first attempts at gene editing occurred in the 1980's. Subsequently, many researchers tried to develop methods to edit specific genes. Here, we review the history of genome editing and improvements in genome editing tools. In the last two decades, genome editing tools have been applied in basic sciences, the bio-industry, and therapeutics. We provide examples in which genome editing tools have been applied to various tasks. Recently, new CRISPR-Cas techniques, such as base and prime editing and anti-CRISPR proteins, have attracted considerable interest. Accordingly, these topics are also reviewed.

Association between worsening renal function severity during hospitalization and low physical function at discharge: a retrospective cohort study of older patients with heart failure and chronic kidney disease from Japan.

BACKGROUND: The number of hospitalized older patients with chronic heart failure, chronic kidney disease, and worsening renal function is rising in Japan. This study aimed to clarify the impact of the severity of worsening renal function during hospitalization on low physical function at discharge of these patients. METHODS: We included 573 consecutive heart failure patients who underwent phase I cardiac rehabilitation. Worsening renal function severity was defined according to elevation during hospitalization of baseline serum creatinine on admission: non-worsening renal function, serum creatinine < 0.2 mg/dL; worsening renal function II/I, serum creatinine ≥ 0.2 to < 0.5 mg/dL; worsening renal function III, and serum creatinine ≥ 0.5 mL/dL. Physical function was measured with the Short Performance Physical Battery. We compared background factors, clinical parameters, pre-hospitalization walking levels, Functional Independence Measure score, and physical function in the three renal function groups. Multiple regression analysis was performed with the Short Performance Physical Battery at discharge as the dependent variable. RESULTS: The final analysis included 196 patients (mean age 82.7 years, male 51.5%) categorized into three groups based on worsening renal function: worsening renal function grade III group (n = 55), worsening renal function grade II/I group (n = 36), and non-worsening renal function group (n = 105). There is no significant difference in walking levels before hospitalization between the three groups, but physical function at discharge was significantly lower in the worsening renal function III group. Moreover, worsening renal function III was an independent factor for low physical function at discharge. CONCLUSION: Worsening of renal function during hospitalization in older patients with heart failure and chronic kidney disease was strongly associated with low physical function at discharge, even after adjusting for other potentially confounding factors, such as pre-hospitalization walking levels, walking start day, and Geriatric Nutrition Risk Index at discharge. Notably, worsening renal function of mild or moderate severity (grade II/I) did not show a significant association with low physical function.

Reconceptualization of Proximal Nail Fold Reconstruction: Skin Grafting over the Nail Plate.

The structure of the periungual area is complicated, resulting in historically difficult morphological reconstruction after trauma or cancer resection. There is also no established standard for its reconstruction; thus, we aimed to use a full-thickness skin graft (FTSG) over the nail plate. Three patients with Bowen disease on their proximal nail fold (PNF) underwent excision with a 2-mm margin preserving the nail matrix, and a temporary wound dressing was applied. The FTSG was harvested from the ipsilateral ulnar wrist joint and placed over the skin defect, including the nail plate. Initially, the FTSG seemed to have shrunken; however, after 3 months, it extended and the PNF had a good color and texture match. Remarkably, the FTSG adhered to the nail plate, and the complex PNF structure appeared well reconstructed. Occasionally, a local flap is used; however, it is limited to small defects and causes a deformity of the periungual structure. In this study, the reconstructed PNF showed good results. We presumed that the bridging phenomenon caused graft survival on the nail plate, and that the presence of stem cells near the nail matrix caused graft extension and eponychium and cuticle regeneration. Specifically, the acquisition of sufficient raw surface around the nail plate and wound preparation after excision resulted in the former, and the nail matrix preservation after excision contributed to the latter. This surgical technique is simple and can therefore be considered a remarkably effective method for periungual area reconstruction to date.

Relation of sedentary behaviour to physical function in phase I cardiac rehabilitation.

Increased sedentary behaviour (SB) is reportedly associated with mortality and morbidity in cardiovascular disease. However, its relation with physical function is not well understood in phase I cardiac rehabilitation (CR). This study aimed to investigate the rate of SB and the relation between SB and physical function among patients participating in phase I CR. This prospective multicentre cohort study enrolled patients participating in CR from October 2020 to July 2022. Patients with probable dementia and difficulty walking alone were excluded. We used sitting SB time as the index of SB and the Short Performance Physical Battery (SPPB) as the index of physical function at discharge. Patients were divided into the low SB group (< 480 min/day) or high SB group (≥ 480 min/day). We analysed and compared the two groups. The final analysis included 353 patients (mean age: 69.6 years, male: 75.6%), of whom 47.6% (168 of 353) were high SB patients. Total sitting SB time was higher in the high SB group versus the low SB group (733.6 ± 155.3 vs 246.4 ± 127.4 min/day, p < 0.001), and mean SPPB score was lower in the high SB group versus the low SB group (10.5 ± 2.4 vs 11.2 ± 1.6 points, p = 0.001). Multiple regression analysis identified SB as an explanatory variable for total SPPB score (p = 0.017). Patients with high SB had significantly lower SPPB scores than those with low SB. These findings underscore the importance of considering SB when improving physical function. Effective strategies to improve physical function can be developed that consider SB in phase I CR.

Effects of social activity participation and trust in the community on the transition of frailty classification in late-stage older adults: a 4-year prospective cohort study.

OBJECTIVES: In Japan, frailty is a major risk factor for requiring long-term care, especially among older adults aged 75 years or older (ie, late-stage older adults). Both physical and social factors (eg, social activities, social support and community trust) are protective factors against frailty. However, few longitudinal studies have examined reversible change or stage improvement in frailty. This study investigated social activity participation and trust in the community that may affect the transition of late-stage older adults' frailty status. DESIGN: A mail-based survey was used to analyse the improvement or deterioration of frailty status (categorised as frailty, pre-frailty and robust) over a 4-year period. Binomial and multinomial logistic regression analyses were conducted; the transition in frailty classification was the dependent variable, while a change in social activity participation and the degree of trust in the community were the independent variables. SETTING: Ikoma City, Nara Prefecture, Japan. PARTICIPANTS: 4249 community-dwelling older adults, aged ≥75 years, not requiring long-term care who completed a follow-up questionnaire from April to May 2016. RESULTS: Adjusting for confounding factors, no significant social factors were detected regarding improvement in frailty. However, an increase in exercise-based social participation was an improvement factor in the pre-frailty group (OR 2.43 (95% CI 1.08 to 5.45)). Conversely, a decrease in community-based social activity was a risk factor in the deterioration from pre-frailty to frailty (OR 0.46 (95% CI 0.22 to 0.93)). In the robust group, increased community-based social activity (OR 1.38 (95% CI 1.00 to 1.90)) was a protective factor against frailty, whereas decreased community trust was a risk factor (OR 1.87 (95% CI 1.38 to 2.52)). CONCLUSIONS: No social factors had a significant influence on the improvement of frailty in late-stage older adults. However, the promotion of exercise-based social participation was found to be important for improvement in the pre-frailty state. TRIAL REGISTRATION NUMBER: UMIN000025621.

Cas9-Geminin and Cdt1-fused anti-CRISPR protein synergistically increase editing accuracy.

Genome editing with CRISPR-Cas9, particularly for therapeutic purposes, should be accomplished via the homology-directed repair (HDR) pathway, which exhibits greater precision than other pathways. However, one of the issues to be solved is that genome editing efficiency with HDR is generally low. A Streptococcus pyogenes Cas9 (SpyCas9) fusion with human Geminin (Cas9-Gem) reportedly increases HDR efficiency slightly. In contrast, we found that regulation of SpyCas9 activity with an anti-CRISPR protein (AcrIIA4) fused to Chromatin licensing and DNA replication factor 1 (Cdt1) significantly increases HDR efficiency and reduces off-target effects. Here, another anti-CRISPR protein, AcrIIA5, was applied, and the combined use of Cas9-Gem and Anti-CRISPR+Cdt1 showed synergistic enhancement of HDR efficiency. The method may be applicable to various anti-CRISPR/CRISPR-Cas combinations.

Relationship between health literacy and physical function of patients participating in phase I cardiac rehabilitation: a multicenter clinical study.

Health literacy (HL) is an important decision factor for health. Both low HL and low physical function cause adverse events in cardiovascular disease patients, but their relationship is not well documented. To clarify the relationship between HL and physical function of patients participating in cardiac rehabilitation and calculate the cutoff value of the 14-item HL scale (HLS) for low handgrip strength, this multicenter clinical study named the Kobe-Cardiac Rehabilitation project for people around the World (K-CREW) was conducted among four affiliated hospitals with patients who underwent cardiac rehabilitation. We used the 14-item HLS to assess HL, and the main outcomes were handgrip strength and Short Physical Performance Battery (SPPB) score. The study included 167 cardiac rehabilitation patients with a mean age of 70.5 ± 12.8 years, and the ratio of males was 74%. Among them, 90 patients (53.9%) had low HL and scored significantly lower in both handgrip strength and SPPB. Multiple linear regression analysis revealed that HL was a determinant factor (ß = 0.118, p = 0.04) for handgrip strength. Receiver operating characteristic analysis revealed the cutoff value of the 14-item HLS for screening for low handgrip strength was 47.0 points, and the area under the curve was 0.73. This study showed that HL was significantly associated with handgrip strength and SPPB in cardiac rehabilitation patients and suggests the possibility of early screening for low HL to improve physical function in cardiac rehabilitation patients with low HL.

Association between forefoot pain and sesamoid rotation angle determined using a weight-bearing plantar ultrasound imaging device.

Forefoot pain is a common symptom for several foot problems. This study aimed to determine whether parameters of forefoot structure (hallux valgus angle (HVA), transverse arch height (TAH) and sesamoid rotation angle (SRA)) are associated with forefoot pain. 547 feet of adult women were divided into two groups: without pain (n = 472) and with pain (n = 75). HVA was measured with a goniometer, TAH and SRA were measured using a weight bearing plantar ultrasound imaging device.Associations between forefoot pain and parameters of forefoot structure were analyzed using the Mann-Whitney U test and univariate and multivariate logistic regression analyses. The intra-rater and inter-rater reliability of the ultrasound images were also tested. SRA was significantly greater in the group with pain compared to the group without pain (p = 0.031) but not HVA (p = 0.057) nor TAH (p = 0.117). The association between forefoot pain and SRA was significant (univariate: p = 0.015 and multivariate p = 0.015), but not between HVA nor TAH. The intra-rater and inter-rater reliability were almost perfect (SRA: ICC1,1 = 0.94, ICC2,1 = 0.91 and TAH: ICC1,1 = 0.88, ICC2,1 = 0.81). We conclude that a higher SRA is related to forefoot pain and should be taken into consideration for assessment of patients with forefoot pain.

Right ventricular mural infective endocarditis after traumatic tricuspid valve regurgitation in a 40-year-old man: A case report.

As the clinical manifestations of traumatic tricuspid valve regurgitation vary according to the extent of tricuspid valve injury, this condition can often go unnoticed and be incidentally discovered. Here, we report the case of a 40-year-old man with patent foramen ovale, in which severe tricuspid regurgitation due to tricuspid valve prolapse was incidentally discovered following blunt chest trauma. Further examination revealed that the prolapse had also caused active right ventricular mural infective endocarditis. The patient had no relevant past medical history of chronic debilitating disease or immunosuppression. After evaluation by the cardiology team, emergent surgical tricuspid valvular repair was successfully performed. Learning objective: Tricuspid valve prolapses resulting from chest trauma may occasionally lead to severe tricuspid regurgitation. Furthermore, this may cause active right ventricular infective endocarditis. In the present case, Staphylococcus aureus was detected in blood cultures, which is usually rapidly progressive and often leads to devastating consequences. Early surgical approach should be considered in cases of infection in the left atrium via patent foramen ovale.

Conservative management of pubertal segmental testicular infarction associated with epididymitis.

Urgent scrotal exploration is performed for segmental testicular infarction (STI) because malignant testicular tumors cannot be ruled out even when STI is suspected on color Doppler ultrasound (US). This report describes the case of a 14-year-old boy who was successfully diagnosed with STI associated with epididymitis using color Doppler US to avoid radical orchiectomy. To our knowledge, this is the first report of STI being diagnosed during puberty and managed using color Doppler US and contrast-enhanced magnetic resonance imaging-guided conservative treatment.