Criteria for storage of cord blood units at Japan's largest cord blood bank.

BACKGROUND AND OBJECTIVES: In Japan, cord blood transplantations exceed those done with adult-sourced unrelated stem cells. This study analyses cord blood (CB) storage criteria to maintain high-quality CB units. MATERIALS AND METHODS: The Kanto-Koshinetsu Cord Blood Bank received 29,795 units from 2014 to 2021, mostly >60 mL, and 5486 (18.4%) were stored as transplantable units. We investigated the mother's gestational period, CB volume, total nucleated cells (TNCs), CD34+ cells, total colony-forming units (CFUs), time from collection to reception and cryopreservation, cell viability, and the reasons for not storing a unit. RESULTS: The average time from collection to reception of 29,795 units was 18.0 h. The most common reason for not storing a CB unit was low cell numbers (pre-processing TNC count <1.2 billion), accounting for 67.9% of the units received. There was no correlation between the CB volume and the CD34+ cell count. The shorter the gestational period, the lower the TNC count, but the higher the CD34+ cell count. There was no correlation between the time from collection to cryopreservation, within a 36-h time limit, and the CD34+ cell recovery rate. CONCLUSION: We could accept units with a TNC count <1.2 billion and a CB volume <60 mL from a gestational period of 38 weeks or less if we did a pre-processing CD34+ cell count. This would secure more units rich in CD34+ cells.

[Repair of Vaginal Cuff Dehiscence after Laparoscopic Radical Cystectomy with Gracilis Myocutaneous Flap].

Vaginal cuff dehiscence after total hysterectomy or total cystectomy had been increasing since laparoscopic or robotic surgery became a common surgery among gynecologists and urologists. A 52-yearold woman underwent laparoscopic radical total cystectomy for muscle invasive bladder carcinoma at Rakuwakai Otowa Hospital. She was emergently admitted with a fist-sized lump protruding from her vagina four months after surgery. Physical examination and her past history on admission disclosed vaginal cuff dehiscence after cystectomy. Computed tomographic scan and magnetic resonance imaging showed no bowel evisceration in the lump. We confirmed that the content of lump was peritoneal tissue and removed it by laparoscopic surgery. Simultaneously, we repaired the vaginal cuff dehiscence with a gracilis myocutaneous flap. There was no subsequent recurrence of vaginal dehiscence or bladder carcinoma in one-year follow-up.

Comparison of mid-term clinical outcomes after acute myocardial infarction in diabetic men between living alone and living together.

Because living alone is associated with an increased risk of type 2 diabetes mellitus in men but not women, living alone may be a risk factor of cardiovascular events after acute myocardial infarction (AMI) in diabetic men. The aim of the present study was to investigate the association between living alone and mid-term clinical outcomes after AMI in diabetic men. We conducted a single center, retrospecitve study. The primary endpoint was the major adverse cardiovascular events (MACE) defined as the composite of all cause death, AMI, and target vessel revascularization. A total of 253 AMI men with diabetes mellitus were included from our hospital records, and divided into the living together group (n = 203) and the living alone group (n = 50). Median follow-up duration was 239 days (Q1: 94 days, Q3: 451 days). A total of 66 MACE was observed during the study period, and Kaplan-Meier curves were constructed to compare the MACE. The MACE was more frequently observed in the living alone group than the living together group (P = 0.041). Multivariate Cox regression analysis revealed that the living alone group was significantly associated with the MACE (Odds ratio: 1.770, 95% confidence interval 1.018-3.077, P = 0.043) after known clinical risk factors. In conclusion, living alone was significantly associated with the mid-term MACE after AMI in diabetic men. It may be important to provide multiple interventions including lifestyle guidance as well as sufficient acute medical care for such high-risk patients.

Residual pulmonary hypertension is associated with clinical outcomes in patients with acute pulmonary thromboembolism.

In patients with acute pulmonary thromboembolism (PTE), the influence of residual pulmonary hypertension (PH) has not been well investigated. The aim of this study was to compare clinical characteristics between patients with and without residual PH, and to investigate the association between residual PH and clinical outcomes after acute phase in acute PTE. We included acute PTE patients who underwent echocardiogram after acute phase between January 2009 and December 2016. These patients were divided into residual PH and non-residual PH groups according to the value of estimated right ventricular systolic pressure (eRVSP) by echocardiogram after acute phase (the residual PH group: eRVSP ≥ 40 mmHg, the non-residual PH group: eRVSP < 40 mmHg). Kaplan-Meier survival curves were applied to investigate whether the residual PH were associated with PTE-related death or recurrent PTE in patients with acute PTE. A total of 49 patients with acute PTE were allocated into the residual PH group (n = 10) and non-residual PH group (n = 39). Median follow-up period for 49 patients was 7 months. The event-free survival rate was significantly lower in the residual PH group as compared with the non-residual PH group (p = 0.003), whereas there was no statistical significance between two groups stratified by initial PH or not (p = 0.97). Residual PH after acute phase was significantly associated with mid-term PTE-related death or recurrent PTE in patients with acute PTE.

Clinical Characteristics and Long-Term Outcomes of Patients with Acute Decompensated Heart Failure with Mid-Range Ejection Fraction.

According to recent guidelines, a new category of patients with heart failure (HF) with mid-range left ventricular ejection fraction (LVEF) (HFmrEF) (LVEF = 40%-49%) has been defined. The purpose of this study was to investigate the clinical characteristics and long-term outcomes of patients with HFmrEF. This was a single-center, retrospective, observational study in which we examined the clinical characteristics and outcomes of 494 consecutive patients with acute decompensated heart failure who were admitted to our institution between January 2014 and December 2016. Of this population, 282 (57.1%), 75 (15.2%), and 137 (48.6%) patients had heart failure with reduced ejection fraction (HFrEF), HFmrEF, and heart failure with preserved ejection fraction (HFpEF), respectively. Ischemic heart disease was the primary etiology in HFmrEF and HFrEF. At the time of discharge, ß-blockers and renin-angiotensin system inhibitors were more frequently prescribed in HFmrEF than in HFpEF. The composite outcome of cardiovascular mortality and HF readmission was significantly lower in HFmrEF than in HFrEF. Further studies are needed to determine the effectiveness of the management of coronary artery disease and cardioprotective medications for HFmrEF.

Right Ventricular Stroke Work Index.

Recently, long-term outcomes of pre-capillary pulmonary hypertension (PH) have been improved, whereas morbidity and mortality are still high because of right-sided heart failure (HF). Right-sided HF is closely related to right ventricular (RV) function and hemodynamics such as RV stroke work index (RVSWI). However, the association between RVSWI and long-term outcomes in pre-capillary PH has not been well investigated. The aims of this study were to compare clinical characteristics between low RVSWI and high RVSWI and to investigate the association between low RVSWI and long-term outcomes in patients with pre-capillary PH. We included patients admitted to diagnose and evaluate PH by right heart catheterization between 2007 and 2015. Patients with pre-capillary PH were divided into two groups according to the median value of RVSWI (low RVSWI group: RVSWI < 19.7 g・m/m2/beat; high RVSWI group: RVSWI ≥ 19.7 g・m/m2/beat). Kaplan-Meier survival curves were applied to investigate whether the low RVSWI were associated with HF death or HF readmission in patients with pre-capillary PH. A total of 36 patients with pre-capillary PH who were diagnosed as pulmonary arterial hypertension (PAH) or chronic thromboembolic PH (CTEPH) were allocated into the low RVSWI group (n = 18) and high RVSWI group (n = 18). The event-free survival rate was significantly lower in the low RVSWI group as compared with the high RVSWI group (P = 0.02). In conclusion, lower RVSWI was significantly associated with HF death or HF readmission in patients with PAH or CTEPH.

High-Intensity Interval Training for Severe Left Ventricular Dysfunction Treated with Left Ventricular Assist Device.

Aerobic training based on anaerobic threshold (AT) is well-known to improve cardiac function, exercise capacity, and long-term outcomes of patients with heart failure. Recent reports suggested that high-intensity interval training (HIIT) for patients with cardiovascular disease may improve cardiopulmonary exercise capacity. We present a 61-year-old male patient of severe left ventricular dysfunction with left ventricular assisted device (LVAD). Following HIIT for 8 weeks, exercise capacity and muscle strength have improved without worsening left ventricular function. Our case showed the possibility that HIIT was feasible and effective even in patients with LVAD.

Clinical Characteristics and Mid-Term Outcomes of Non-Elderly Obese Patients with Acute Decompensated Heart Failure in Japan.

Obesity is a well-known risk factor for cardiovascular diseases including heart failure (HF). However, some literatures suggested better clinical outcomes in obese patients with HF. Since higher body mass index (BMI) levels of HF patients were significantly associated with younger age, the impact of obesity on clinical outcomes in non-elderly HF patients should be elucidated.Consecutive 155 non-elderly acute decompensated HF patients (< 60-year-old) who admitted to our institution between 2009 and 2013 were included. Those patients were divided into the two groups according to the BMI: the obesity group (BMI ≥ 25 kg/m2, n = 81) and the non-obesity group (BMI < 25 kg/m2, n = 74). The primary composite outcome of this study was defined as re-admission due to HF and all-cause death.The primary composite outcome was less frequently observed in the obesity group as compared with the non-obesity group (Hazard ratio [HR] 0.50, 95% confidence interval [CI] 0.26-0.95, P = 0.03). Re-admission due to HF was significantly less in the obesity group than in the non-obesity group (HR 0.44, 95% CI 0.23-0.86, P = 0.02), whereas all-cause death was not significantly different between the groups (P = 0.44).The mid-term outcomes in non-elderly HF patients with obesity were better as compared with non-elderly HF patients without obesity, which supports obesity paradox in this specific population.

A Case of Pulmonary Hypertension Associated with Idiopathic Hypereosinophilic Syndrome.

Hypereosinophilic syndrome (HES) is characterized by multi-organ damage that is associated with tissue hypereosinophilia. A persistently elevated eosinophilic count is also required for the diagnosis of HES. Although HES affects various organs, damage to pulmonary artery is rarely reported. We present a case of a 39-year-old man who was diagnosed with pulmonary hypertension (PH) associated with idiopathic HES. Although the pulmonary arterial hypertension specific drugs including intravenous epoprostenol could not control his PH, corticosteroid was effective for both hypereosinophilia and PH. Our case suggests the importance of steroid therapy as well as specific drugs for pulmonary arterial hypertension in the treatment of PH associated with HES.

Evaluation of an in vitro muscle contraction model in mouse primary cultured myotubes.

To construct an in vitro contraction model with the primary cultured myotubes, we isolated satellite cells from the mouse extensor digitorum longus. Differentiated myotubes possessed a greater number of sarcomere assemblies and higher expression levels of myosin heavy chain, cytochrome c oxidase IV, and myoglobin than in C2C12 myotubes. In agreement with these results regarding the sarcomere assemblies and protein expressions, the primary myotubes showed higher contractile activity stimulated by the electric pulses than that in the C2C12 myotubes. These data suggest that mouse primary myotubes will be a valuable research tool as an in vitro muscle contraction model.

A fragmented form of annexin A1 is secreted from C2C12 myotubes by electric pulse-induced contraction.

The main function of annexin A1 (ANXA1), a member of the annexin superfamily, is to bind to cellular membranes in a Ca(2+)-dependent manner. In skeletal muscle, ANXA1 is thought to be involved in the repair of damaged membrane tissue and in the migration of muscle cells. We hypothesized that ANXA1 is one of the myokines secreted during muscle contractions to accelerate the repair of cell damage after contraction. Here we performed cell contractions by electric pulse stimulation; the results revealed that a fragmented form of ANXA1 was cleaved by calpain and selectively secreted from skeletal muscle cells by contraction. We therefore realized that muscle-contraction-induced calpain-dependent ANXA1 fragmentation has a wound-healing effect on damaged cells. This suggested that not the intact form but rather fragmented ANXA1 is a contraction-induced myokine.

Clostridium oryzae sp. nov., from soil of a Japanese rice field.

An obligately anaerobic bacterial strain designated KC3(T) was isolated from a rice straw-degrading culture, for which soil of a Japanese rice field was used as the inoculum. Cells of strain KC3(T) were determined to be non-cellulolytic, Gram-stain-positive, non-motile, ellipsoidal, spore-forming rods, 0.8-1×4-25 µm. Endospores were formed at a terminal position in elongated cells (12-25 µm, mean 15 µm). The temperature range for growth was 20-50 °C, with an optimum at 37 °C. The pH range for growth was 5.0-7.5, with an optimum at pH 6.0 (slightly acidophilic). Strain KC3(T) fermented cellobiose to lactate, butyrate, acetate, formate, hydrogen and carbon dioxide. The major cellular fatty acids (>10 %) were C14 : 0, C16 : 0 and C19 : 0 cyclo 11,12 dimethylacetal. The DNA G+C content of strain KC3(T) was 37.5 mol%. 16S rRNA gene sequence analysis revealed that strain KC3(T) shared low sequence similarity (<93 %) with type strains of the genus Clostridium sensu stricto (Clostridium rRNA cluster I). Analyses of the DNA gyrase A and ATP synthase beta subunit sequences supported the affiliation of strain KC3(T) to the genus Clostridium sensu stricto. The evidence presented here indicates that strain KC3(T) represents a novel species of the genus Clostridium, for which the name Clostridium oryzae sp. nov. is proposed. The type strain of Clostridium oryzae is KC3(T) ( = DSM 28571(T) = NBRC 110163(T)).

Transverse and longitudinal right ventricular fractional parameters derived from four-chamber cine MRI are associated with right ventricular dysfunction etiology.

Studies of the usefulness of transverse right ventricular (RV) shortening are limited. We retrospectively analyzed the CMR images of 67 patients (age: 50.8 ± 19.0 years; men: 53.7%; Control: n = 20, Overloaded RV (atrial septal defect): n = 15, Constricted RV (pericarditis): n = 17, Degenerated RV (arrhythmogenic right ventricular cardiomyopathy): n = 15) (all enrolled consecutively for each disease) in a single center. We defined RV longitudinal (fractional longitudinal change: FLC) and transverse (fractional transverse change: FTC) contraction parameters. We assessed the FTC/FLC (T/L) ratio on four-chamber cine CMR views and compared the four groups regarding the fractional parameters. FTC had a stronger correlation (R2 = 0.650; p < 0.001) with RV ejection fraction than that with FLC (R2 = 0.211; p < 0.001) in the linear regression analysis. Both FLC and FTC were significantly lower in the Degenerated RV and Constricted RV groups compared with those in the Control and Overloaded RV groups. The T/L ratio was significantly lower in the Degenerated RV group (p = 0.008), while the Overloaded RV (p = 0.986) and Constricted RV (p = 0.582) groups had preserved T/L ratios, compared with the Control group. Transverse shortening contributes to RV function more significantly compared with longitudinal contraction. Impaired T/L ratios may reflect RV myocardial degeneration. RV fractional parameters may help precisely understand RV dysfunction.

TIMP-3 recruits quiescent hematopoietic stem cells into active cell cycle and expands multipotent progenitor pool.

Regulating transition of hematopoietic stem cells (HSCs) between quiescent and cycling states is critical for maintaining homeostasis of blood cell production. The cycling states of HSCs are regulated by the extracellular factors such as cytokines and extracellular matrix; however, the molecular circuitry for such regulation remains elusive. Here we show that tissue inhibitor of metalloproteinase-3 (TIMP-3), an endogenous regulator of metalloproteinases, stimulates HSC proliferation by recruiting quiescent HSCs into the cell cycle. Myelosuppression induced TIMP-3 in the bone marrow before hematopoietic recovery. Interestingly, TIMP-3 enhanced proliferation of HSCs and promoted expansion of multipotent progenitors, which was achieved by stimulating cell-cycle entry of quiescent HSCs without compensating their long-term repopulating activity. Surprisingly, this effect did not require metalloproteinase inhibitory activity of TIMP-3 and was possibly mediated through a direct inhibition of angiopoietin-1 signaling, a critical mediator for HSC quiescence. Furthermore, bone marrow recovery from myelosuppression was accelerated by over-expression of TIMP-3, and in turn, impaired in TIMP-3-deficient animals. These results suggest that TIMP-3 may act as a molecular cue in response to myelosuppression for recruiting dormant HSCs into active cell cycle and may be clinically useful for facilitating hematopoietic recovery after chemotherapy or ex vivo expansion of HSCs.

Assessment of dysarthria with Frenchay dysarthria assessment (FDA-2) in patients with Duchenne muscular dystrophy.

PURPOSE: The purpose of this study was to test the psychometric properties of the Japanese version of Frenchay Dysarthria Assessment (FDA-2) and to use this tool to describe the features of speech in patients with Duchenne muscular dystrophy (DMD). MATERIALS AND METHODS: The Japanese version of FDA-2 was culturally adapted, and reliability and validity were examined in 22 and 50 patients, respectively. The Japanese version of FDA-2 was administered to 51 patients with DMD. Multiple regression analysis was performed to identify factors related to FDA-2 scores. RESULTS: Inter-/intra-rater reliabilities (ICCs) and internal consistency (Cronbach's α) for total scores were 0.76, 0.97, and 0.94 respectively. For construct validity, two-way ANOVA showed a significant interaction between the disorders and FDA-2 sections (p < 0.05). In DMD patients, the item of tongue at rest was most severely affected, reflecting tongue hypertrophy. Multiple regression analysis identified age, swallowing status, and ventilator use as significantly related. CONCLUSIONS: The results showed that the Japanese version of FDA-2 has satisfactory reliability and validity. The present study demonstrated the features of dysarthria and related factors in patients with DMD.Implications for rehabilitationIn Duchenne muscular dystrophy (DMD), an absent or defective dystrophin protein causes progressive weakness of respiratory and oropharyngeal muscles, both of which are crucial contributors to speech production.This study shows that the Japanese version of FDA-2 has satisfactory reliability and validity compared to original version.The Japanese version of FDA-2 characterizes dysarthria in patients with DMD in this cohort.

Association between class of foundational medication for heart failure and prognosis in heart failure with reduced/mildly reduced ejection fraction.

We clarified the association between changes in the number of foundational medications for heart failure (FMHF) during hospitalization for worsening heart failure (HF) and post-discharge prognosis. We retrospectively analyzed a combined dataset from three large-scale registries of hospitalized patients with HF in Japan (NARA-HF, WET-HF, and REALITY-AHF) and patients diagnosed with HF with reduced or mildly reduced left ventricular ejection fraction (HFr/mrEF) before admission. Patients were stratified by changes in the number of prescribed FMHF classes from admission to discharge: angiotensin-converting enzyme inhibitors or angiotensin receptor blockers, beta-blockers, and mineralocorticoid receptor blockers. Primary endpoint was the combined endpoint of HF rehospitalization and all-cause death within 1 year of discharge. The cohort comprised 1113 patients, and 482 combined endpoints were observed. Overall, FMHF prescriptions increased in 413 (37.1%) patients (increased group), remained unchanged in 607 (54.5%) (unchanged group), and decreased in 93 (8.4%) (decreased group) at discharge compared with that during admission. In the multivariable analysis, the increased group had a significantly lower incidence of the primary endpoint than the unchanged group (hazard ratio 0.56, 95% confidence interval 0.45-0.60; P < 0.001). In conclusion, increase in FMHF classes during HF hospitalization is associated with a better prognosis in patients with HFr/mrEF.

SHD1 is a novel cytokine-inducible, negative feedback regulator of STAT5-dependent transcription.

STAT5 is a critical mediator of a variety of cytokine signaling whose transcriptional activity is regulated by associating with various proteins. During a search for STAT5-interacting proteins, we identified SHD1, a mammalian homologue of yeast gene Sac3, as a potential interacter. SHD1 was localized in the nucleus, and induced by cytokines that activate STAT5, such as erythropoietin, interleukin-2 (IL-2), or IL-3. SHD1 interacted specifically with STAT5A and STAT5B, and interestingly, it specifically repressed STAT5-dependent transcription in vitro without affecting the stability or phosphorylation of STAT5 protein. Gene disruption study revealed that T, B, or bone marrow cells from mice lacking SHD1 were hyperresponsive to T-cell-receptor engagement, or stimulation with various STAT5-activating cytokines. These results suggest that SHD1 is a novel cytokine-inducible negative feedback regulator of STAT5.

Cord blood index predicts engraftment and early non-relapse mortality in adult patients with single-unit cord blood transplantation.

How to select optimal cord blood (CB) remains an important clinical question. We developed and validated an index of CB engraftment, the cord blood index (CBI), which uses three weighted variables representing cell doses and HLA mismatches. We modeled the neutrophil engraftment time with competing events by random survival forests for competing risks as a function of the predictors: total nucleated cells, CD34, colony-forming units for granulocytes/macrophages, and the number of HLA mismatches at the antigen and allele levels. The CBI defined three groups that had different neutrophil engraftment rates at day 30 (High, 83.7% [95% CI, 79.2-88.1%]; Intermediate, 77.0% [95% CI, 73.7-80.2%]; Low, 68.4% [95% CI, 63.6-73.2%]), platelet engraftment rates at day 60 (High, 70.4% [95% CI, 64.9-75.9%]; Intermediate, 62.3% [95% CI, 58.5-66.0%]; Low, 49.3% [95% CI, 44.2-54.5%]), and non-relapse mortality at day 100 (High, 14.1% [95% CI, 9.9-18.3%]; Intermediate, 16.4% [95% CI, 13.5-19.3%]; Low, 21.3% [95% CI, 17.1-25.5%]). This novel approach is clinically beneficial and can be adopted immediately because it uses easily obtained pre-freeze data of CB.

Roundabout 4 is expressed on hematopoietic stem cells and potentially involved in the niche-mediated regulation of the side population phenotype.

Roundabout (Robo) family proteins are immunoglobulin-type cell surface receptors that are expressed predominantly in the nervous system. The fourth member of this family, Robo4, is distinct from the other family members in that it is expressed specifically in endothelial cells. In this study, we examined the expression of Robo4 in hematopoietic stem cells (HSCs) and its possible role in HSC regulation. Robo4 mRNA was specifically expressed in murine HSCs and the immature progenitor cell fraction but not in lineage-positive cells or differentiated progenitors. Moreover, flow cytometry showed a correlation between higher expression of Robo4 and immature phenotypes of hematopoietic cells. Robo4(high) hematopoietic stem/progenitor cells presented higher clonogenic activity or long-term repopulating activity by colony assays or transplantation assays, respectively. A ligand for Robo4, Slit2, is specifically expressed in bone marrow stromal cells, and its expression was induced in osteoblasts in response to myelosuppressive stress. Interestingly, overexpression of Robo4 or Slit2 in HSCs resulted in their decreased residence in the c-Kit(+)Sca-1(+)Lineage(-)-side population fraction. These results indicate that Robo4 is expressed in HSCs, and Robo4/Slit2 signaling may play a role in HSC homeostasis in the bone marrow niche.