A culture method with berbamine, a plant alkaloid, enhances CAR-T cell efficacy through modulating cellular metabolism.

Memory T cells demonstrate superior in vivo persistence and antitumor efficacy. However, methods for manufacturing less differentiated T cells are not yet well-established. Here, we show that producing chimeric antigen receptor (CAR)-T cells using berbamine (BBM), a natural compound found in the Chinese herbal medicine Berberis amurensis, enhances the antitumor efficacy of CAR-T cells. BBM is identified through cell-based screening of chemical compounds using induced pluripotent stem cell-derived T cells, leading to improved viability with a memory T cell phenotype. Transcriptomics and metabolomics using stem cell memory T cells reveal that BBM broadly enhances lipid metabolism. Furthermore, the addition of BBM downregulates the phosphorylation of p38 mitogen-activated protein kinase and enhanced mitochondrial respiration. CD19-CAR-T cells cultured with BBM also extend the survival of leukaemia mouse models due to their superior in vivo persistence. This technology offers a straightforward approach to enhancing the antitumor efficacy of CAR-T cells.

Effective and stable gene transduction in rhesus macaque iPSCs capable of T-lineage differentiation utilizing the piggyBac system.

Introduction: Genetically modified human induced pluripotent stem cell (iPSC)-based regenerative medicine has substantial potential in the treatment of refractory human diseases. Thus, preclinical studies on the safety and efficacy of these products are essential. Non-human primate (NHP) models such as the rhesus macaque are highly similar to humans in terms of size, lifespan, and immune system, rendering them superior models. However, effective gene transduction in rhesus macaque iPSCs (Rh-iPSCs) remains challenging. In this study, we investigated the effective gene transduction into Rh-iPSCs and its effect on differentiation efficiency. Methods: We established a gene transduction method using the piggyBac transposon vector system. Gene transduced Rh-iPSCs were analyzed for undifferentiated markers. We did teratoma assay to check pluripotency. Gene transduced Rh-iPSCs were differentiated into hematopoietic stem and progenitor cells (HSPCs) and T-cell lineage cells. Additionally, gene transduced Rh-iPSCs were compared the differentiation efficiency with parental Rh-iPSCs. Results: We could establish a gene transduction method using the piggyBac transposon vector system, demonstrating high efficiency and stable transgene expression in Rh-iPSCs. These Rh-iPSCs maintained long-term gene expression while expressing undifferentiated markers. Teratoma assay indicated that these Rh-iPSCs had pluripotency. These Rh-iPSCs could differentiate into HPSCs and T cells that express transgenes. These Rh-iPSCs can differentiate into hematopoietic stem cells and T cells that express transgenes. No significant differences in efficiency of differentiation were observed between parental Rh-iPSCs and these Rh-iPSCs. Conclusions: These results indicate that the piggyBac transposon vector is an excellent gene transfer tool for rhesus macaque iPSCs and could contribute to the advancement of preclinical studies using rhesus macaque iPSCs.

Short term-efficacy and tolerability of Rheocarna, a novel direct hemoperfusion adsorptive column, for chronic limb-threatening ischemia in dialysis patients: A single-center case series.

INTRODUCTION: With population aging and lifestyle changes, the number of patients with chronic limb-threatening ischemia (CLTI) is increasing, and refractory or recurrent lesions are more common, especially in chronic dialysis patients. In March 2021, a new type of adsorptive cellulose bead column immobilized with dextran sulfate and L-tryptophan for direct hemoperfusion (DHP) was approved by Japan's medical insurance system as a treatment for CLTI. METHODS: We retrospectively analyzed 17 cases of CLTI in dialysis patients treated with DHP using the novel column (Rheocarna) (DHP-R) at our hospital from May 2021 to October 2022. The short-term of efficacy of DHP-R was judged qualitatively by the foot care team every 2 weeks based on the assessment of skin color, warmth, ulcer epithelialization or shrinkage of the ulcer area, and foot pain. The final judgment of efficacy was made after the final DHP-R session. RESULTS: The median age of patients was 66 years, the median dialysis duration was 10 years, 15 cases (88%) were male, and 15 cases (88%) had diabetes. The median total number of sessions was eight. In comparing the groups in which DHP-R was effective and ineffective, there was no significant difference in any factors including patient background data (i.e., age, diabetes, low-density lipoprotein cholesterol, hemoglobin, dialysis duration, etc.), type of anticoagulants, and presence of episodes of blood pressure drop or circuit clotting during session. Three cases with symptomatic hypotension during the session and two cases with circuit clotting that did not improve with increased heparin dose all resolved immediately after changing the anticoagulant from heparin to nafamostat mesylate (NM). CONCLUSION: Identification of patients' characteristics in which DHP-R is favorable and some reliable index that allow a rapid decision to continue DHP-R are needed. In addition, validating whether the use of NM as anticoagulant affects the efficacy of DHP-R for CTLI treatment remains a challenge to resolve.

Successful Use of Granulocyte and Monocyte Adsorptive Apheresis in a Patient with Post-COVID-19 Multisystem Inflammatory Syndrome in Adults.

Multisystem inflammatory syndrome in adults (MIS-A) is a life-threatening disease that can develop weeks after coronavirus disease 2019 (COVID-19). MIS-A symptoms include multiorgan involvement, especially gastrointestinal tract and heart involvement, and Kawasaki disease-like symptoms. We herein report a 44-year-old Japanese man with MIS-A who had contracted COVID-19 five weeks ago and went into shock after acute gastroenteritis, acute kidney injury, and Kawasaki disease-like symptoms. Methylprednisone pulse and high-dose intravenous immunoglobulin resulted in recovery of shock and his renal function, but diffuse ST-segment elevation on electrocardiography and pericardial effusion with a fever emerged after therapy. Additional granulocyte-monocyte adsorptive apheresis successfully ameliorated the cardiac involvement.

Development of IgA vasculitis with severe glomerulonephritis after COVID-19 vaccination: a case report and literature review.

With the worldwide spread of the COVID-19 vaccine program during the COVID-19 pandemic, the numbers of reported cases with new-onset or relapsed kidney disease/vasculitis such as minimal change nephrotic syndrome, immunoglobulinA (IgA) nephropathy, and IgA vasculitis (IgAV) that developed after COVID-19 vaccination are increasing. We present the case of a 67-year-old Japanese woman who developed IgAV with purpura on her extremities and trunk in the evening of the day that she received the second dose of the Pfizer-BioNTech COVID-19 vaccine. She subsequently presented with acute kidney injury and nephrotic syndrome, and a kidney biopsy performed 14 days after the second vaccination showed diffuse mesangial and endocapillary glomerulonephritis with necrotizing crescent formation, accompanied by IgA deposition. One steroid pulse plus four administrations of a monthly intravenous cyclophosphamide injection were applied, followed by oral azathioprine during oral steroid tapering. Her response to this treatment was unsatisfactory and intractable for some time. Eventually, her renal function improved and nephrotic syndrome was resolved, while microscopic hematuria and proteinuria at ~ 1 g/gCr remained at 6 months post-vaccination. Unlike the previous milder renal-involved IgAV cases following COVID-19 vaccination, our patient's case presented severe glomerulonephritis and took a long time to recover despite intensive initial immunosuppressive treatment.

The association between subjective health perception and lifestyle factors in Shiga prefecture, Japan: a cross-sectional study.

BACKGROUND: The Ministry of Health has reported that in Japan, the Shiga prefecture has the highest life expectancy. Subjective health perception is a predictive indicator of mortality. For this study, we examined the association between subjective health perception and multiple lifestyle factors. METHODS: Data were obtained from the 2015 Health and Nutrition Survey in Shiga prefecture. The analytic sample comprised 6057 adults aged 20 or older. Information on subjective health perception and lifestyle behaviors was obtained from a self-administered questionnaire. As for subjective health perception, participants were divided into 2 groups: (1) Excellent or Good and (2) Average, Poor, or Very Poor. A 1-day dietary survey was also administered. The health behaviors score (HBS) was calculated based on 5 factors: consuming a healthy diet, never smoking, low-risk alcohol drinking, regular exercise, and moderate sleep duration. HBS scores ranged from 0 to 5. Multiple logistic regression was used to calculate the sex-, age- BMI- and energy intake-adjusted odds ratios (ORs) of poor subjective health across HBS, with 0 points as the reference. RESULTS: Among all participants, 2397 (39.6%) individuals were classified into the good subjective health group. Participants with an HBS of 3 (OR 0.59, 95% CI 0.37-0.96), 4 (OR 0.40, 95% CI 0.24-0.65) or 5 (OR 0.33, 95% CI 0.19-0.59) had a lower OR of rating themselves as being average/poor health compared with those having zero. The association with a higher HBS was remarkable (p for trend: < 0.001). Additional analyses revealed that the combinations including regular exercise were particularly associated with a lower risk of subjective average/poor health. CONCLUSIONS: This study showed that the higher the number of healthy lifestyle factors, the lower risk of subjective average/poor health. Combinations of healthy lifestyle factors, especially those involving exercise, suggest good subjective health for individuals living in the Shiga prefecture.

The Association between Combined Lifestyle Factors and All-Cause and Cause-Specific Mortality in Shiga Prefecture, Japan.

BACKGROUND: To decrease mortality, the benefit of combined healthy lifestyles has been suggested but is still unclear, especially for cause-specific mortality. We examined the relationship between combined lifestyle factors and all-cause and cause-specific mortality in Shiga prefecture, Japan. METHODS: This was an ecological study of 19 municipalities, using the data from the 2008-2012 standard mortality ratio (SMR) reported by the Ministry of Health and Welfare and the 2015 Health and Nutrition Survey in Shiga prefecture. The health behaviors score was calculated based on five factors (ranging from 0 to 5): diet quality (assessed adherence to dietary reference intake for Japanese), smoking, alcohol drinking, regular exercise, and sleep duration. In the multiple linear regression, the relationships between the health behaviors score and SMR of all-cause, cancer, heart diseases, and cerebrovascular diseases were estimated by sex. RESULTS: The health behaviors score was negatively associated with the cancer SMR in women (ß = -0.968, p = 0.011). For other causes, no significant association was found for either sex. A greater proportion of those who never smoked (ß = -0.780, p = 0.016) and those who had a higher quality diet (ß = -0.703, p = 0.048) were associated with lower cancer SMR in women. Women's intake of some micronutrients, particularly fruits, was higher than men. This study suggests that a combination of health behaviors, especially never smoking and high-quality diet intake are associated with lower cancer SMR in women and could be helpful in prolonging life expectancy.

The precise long-term outcomes of adult IgA nephropathy by mail questionnaires: Better renal survival compared to earlier cohort studies.

The estimated 20-year renal survival rate of immunoglobulin A (IgA) nephropathy is approx. 60%, but it is difficult to determine the 'big picture' for IgA nephropathy because a biopsy is essential for its diagnosis. Here we attempted to determine the longer and more precise renal prognosis of IgA nephropathy. We examined 310 patients with primary IgA nephropathy. Using the patients' clinical records and histological reports from our hospital and other clinics, we surveyed their renal prognoses and treatments within 1 year post-biopsy, and we sent questionnaires to the patients who had stopped visiting any hospital. We set renal death as the primary endpoint and analyzed factors related to renal death. The total patient cohort was 267: 159 males, 108 females; average age at biopsy, 37.7 years; average estimated glomerular filtration rate (eGFR), 69.7 mL/min/1.73m2; urinary protein, 1.3 g/day. The mean follow-up duration was prolonged to 13.8±8.9 years (vs. 9.2±8.5 years using only medical records). The 10- and 20-year follow-up rates were 61.7% and 27.3%. The 10-, 20-year renal survival rates were 83.6% and 72.5%. Lower eGFR, hypertension, and smoking were revealed as factors independently related to renal death. To study survival of relatively benign diseases such as IgA nephropathy, longer survival rate was affected by many censoring cases. The results regarding the long-term renal prognoses of IgA nephropathy patients (including those with a mild phenotype) obtained by our analysis of a questionnaire sent to the patients provided more precise and longer-term prognoses compared to earlier studies.

The Association with Dietary Patterns and Risk of Anemia in Japanese Elderly.

Anemia is a risk factor for frailty. Although various factors can reduce the risk of anemia, information on dietary contributions is limited. This study aimed to evaluate the association between anemia and eating patterns in Japanese elderly. Our study included 6,864 elderly individuals, aged ≥65 y, whose data were obtained from the 2010-2015 National Health and Nutritional Survey, Japan. Anemia was defined as whole blood hemoglobin concentrations <13 g/dL in men, and <12 g/dL in women. We classified eating patterns into 33 food groups, using food items obtained from dietary records, and identified three dietary patterns using principal component analysis method. Participants were assigned and divided into quartiles according to pattern-specific factor scores. We calculated adjusted odds ratios (ORs) for anemia using multivariate logistic regression, and assigned the first quartile (minimum) as the reference. The overall prevalence of anemia was 17.1%. The meats and vegetables eating pattern was characterized by red meat and other vegetables, the Japanese eating pattern by rice and salted or pickled vegetables, and the fruits and vegetables eating pattern by fruits and green-yellow vegetables. Participants in the highest meats and vegetables eating pattern quartile had a 20% lower risk of anemia than those in the lowest quartile (OR 0.81; 95% confidence interval (CI) 0.66-1.00); the Japanese eating pattern participants had a 20% higher risk of anemia (OR 1.28; 95% CI 1.06-1.53). The fruits and vegetables eating pattern participants had no significant change in their risk of anemia. This study may indicate that consuming a high red meat diet prevents anemia in elderly Japanese individuals.

Fish and meat intakes and prevalence of anemia among the Japanese elderly.

BACKGROUND AND OBJECTIVES: Information about an association between animal food intakes and risk of anemia is still limited. This study aimed to investigate the association between fish and meat intake and anemia risk in the Japanese elderly. METHODS AND STUDY DESIGN: A nationally representative sample of 6,469 aged 65 years and over was obtained from pooled data of annual National Health and Nutritional Survey in Japan during 2002-2011. Anemia was defined as hemoglobin concentrations <13.0 g/dL in males and <12.0 g/dL in females. Logistic regression analysis, with the lowest intake tertile as the reference, was applied to estimate anemia risk for each nutrient and food group. RESULTS: After adjustment for putative confounding factors, males in the highest tertile of animal protein intake had significantly lower risk of anemia than those in the lowest tertile (odds ratio (OR): 0.77; 95% confidence interval (CI): 0.63, 0.95; p for trend=0.017). These associations were not seen in females (OR: 0.72, 95% CI: 0.49, 1.06; p for trend=0.100). Multivariate analyses revealed that anemia risk (OR: 0.80; 95% CI: 0.65, 0.97; p for trend =0.002) was lower for males in the highest tertile of fish intake than in the lowest tertile; this effect was also observed for females (OR: 0.64; 95% CI: 0.45, 0.92; p for trend =0.014). In both sexes, the highest tertile of meat intake was not associated with lower anemia risk in the multivariate-adjusted models. CONCLUSIONS: The current cross-sectional study in Japanese elderly males suggests that higher animal protein, specifically the high protein content of fish may be associated with a lower prevalence of anemia.

Associations of complementation group, ALDH2 genotype, and clonal abnormalities with hematological outcome in Japanese patients with Fanconi anemia.

Fanconi anemia (FA) is a genetically and clinically heterogeneous disorder that predisposes patients to bone marrow failure (BMF), myelodysplastic syndromes (MDS), and acute myeloid leukemia (AML). To study which genetic and phenotypic factors predict clinical outcomes for Japanese FA patients, we examined the FA genes, bone marrow karyotype, and aldehyde dehydrogenase-2 (ALDH2) genotype; variants of which are associated with accelerated progression of BMF in FA. In 88 patients, we found morphologic MDS/AML in 33 patients, including refractory cytopenia in 16, refractory anemia with excess blasts (RAEB) in 7, and AML in 10. The major mutated FA genes observed in this study were FANCA (n = 52) and FANCG (n = 23). The distribution of the ALDH2 variant alleles did not differ significantly between patients with mutations in FANCA and FANCG. However, patients with FANCG mutations had inferior BMF-free survival and received hematopoietic stem cell transplantation (HSCT) at a younger age than those with FANCA mutations. In FANCA, patients with the c.2546delC mutation (n = 24) related to poorer MDS/AML-free survival and a younger age at HSCT than those without this mutation. All patients with RAEB/AML had an abnormal karyotype and poorer prognosis after HSCT; specifically, the presence of a structurally complex karyotype with a monosomy (n = 6) was associated with dismal prognosis. In conclusion, the best practice for a clinician may be to integrate the morphological, cytogenetic, and genetic data to optimize HSCT timing in Japanese FA patients.

Enhancing T Cell Receptor Stability in Rejuvenated iPSC-Derived T Cells Improves Their Use in Cancer Immunotherapy.

Limited T cell availability and proliferative exhaustion present major barriers to successful T cell-based immunotherapies and may potentially be overcome through the use of "rejuvenated" induced pluripotent stem cells derived from antigen-specific T cells (T-iPSCs). However, strict antigen specificity is essential for safe and efficient T cell immunotherapy. Here, we report that CD8αß T cells from human T-iPSCs lose their antigen specificity through additional rearrangement of the T cell receptor (TCR) α chain gene during the CD4/CD8 double positive stage of in vitro differentiation. CRISPR knockout of a recombinase gene in the T-iPSCs prevented this additional TCR rearrangement. Moreover, when CD8αß T cells were differentiated from monocyte-derived iPSCs that were transduced with an antigen-specific TCR, they showed monoclonal expression of the transduced TCR. TCR-stabilized, regenerated CD8αß T cells effectively inhibit tumor growth in xenograft cancer models. These approaches could contribute to safe and effective regenerative T cell immunotherapies.

Oral Glucose Tolerance and Tryptophan Metabolism in Non-Obese and Non-Insulin-Dependent Diabetic Goto-Kakizaki Rats Fed High-Tryptophan Diets.

We investigated oral glucose tolerance and tryptophan (Trp) metabolism in non-obese and non-insulin-dependent diabetic Goto-Kakizaki (GK) rats fed high-Trp diets. Five-week-old male Wistar and GK rats were fed a 20% casein diet (control diet) or the same diet supplemented with 1%, 2%, 3%, or 5% Trp for 58 d. Oral glucose tolerance tests were performed on Days 14 and 28 of the experimental period. Urine as well as livers and blood were collected on the last day of the experiment. The glucose concentration and the amount of Trp metabolites were measured. On Day 14 of the experiment, the incremental blood glucose concentrations integrated over a period of 2 h (ΔAUC0-2h) of blood glucose in rats fed the 3% and 5% Trp diets had decreased by 13% and 18%, respectively, compared with that of the control-GK rats. However, no significant differences were found in the rats fed +1% or +2% Trp diets compared with control-GK rats. On Day 28, there were no significant differences found in the ΔAUC0-2h of blood glucose levels in any group including the control-GK group. On the last day, the concentrations of plasma glucose, total cholesterol, and triglyceride did not show differences in any group. There were no specific phenomena observed in the metabolism of Trp in GK rats even when fed an excess of Trp, compared with that of Wistar rats. Oral Trp administration and its continuous use may not improve blood glucose levels in type 2 diabetic rats.

Anaemia is an essential complication of ANCA-associated renal vasculitis: a single center cohort study.

BACKGROUND: Anaemia is a common complication of patients with antineutrophil cytoplasmic antibody (ANCA)-associated renal vasculitis. Nevertheless, the cause and degree of such cases of anaemia have not been elucidated in detail. We aimed to investigate the prevalence, cause, pathogenesis of anaemia and the impact of anaemia on prognosis in patients with ANCA-associated renal vasculitis. METHODS: We identified 45 patients with ANCA-associated renal vasculitis that were clinically and/or histologically diagnosed and treated from 2003 to 2014 at University of Tsukuba Hospital. The relationships between anaemia and various clinicopathological findings were evaluated. RESULTS: At the time of diagnosis of ANCA-associated renal vasculitis, all patients showed anaemia, with a mean haemoglobin level of 7.5 ± 1.3 g/dL. Renal anaemia was diagnosed in 92% of patients, anaemia of chronic disease (ACD) in 56%, and anaemia due to hemorrhage in 20%. Next, the patients were divided into two groups according to anaemia severity: minimum haemoglobin (min Hb) < 7.5 (n = 24) and min Hb ≥ 7.5 (n = 21). A comparison of baseline characteristics showed that serum albumin, maximum serum creatinine, minimum estimated glomerular filtration rate (eGFR), serum cystatin C, and the area of tubulointerstitial damage were significantly different between the haemoglobin groups (p < 0.05). No significant intergroup differences were observed in iron-related or inflammation-related data. With regard to the relationship between anaemia severity and prognosis, patients in the min Hb < 7.5 group tended to have a lower eGFR. Anaemia severity was associated with markedly lower survival (Log-rank test, p = 0.03). CONCLUSIONS: In this cohort of patients with ANCA-associated renal vasculitis, all subjects exhibited anaemia. In regard to the cause and pathogenesis, the most prevalent form of anaemia was renal anaemia, not ACD, and a potential reason for the high prevalence of anaemia in our cohort may have been the interaction between renal anaemia and ACD. Moreover, anaemia severity was significantly associated with the degree of renal dysfunction and life prognosis.

An objective evaluation of nocturnal cough count and cough pattern in children with psychogenic cough.

Two patients with a chronic, barking cough were diagnosed with psychogenic cough. Using an original cough counter we studied the nocturnal cough count and pattern. While the number of coughs when awake was extremely high for both patients, the number of coughs was remarkably reduced during sleep, similar to an exacerbation of asthma. Moreover, the properties of the coughs when awake were clearly different from those of coughs during sleep. In conclusion, an objective examination using a cough counter was useful for the diagnosis, treatment and management of psychogenic cough.

Renal compensatory adaptation for water handling in a patient with adipsic diabetes insipidus after clipping of a ruptured aneurysm of the anterior communicating artery
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A 38-year-old Japanese man who had undergone clipping surgery for a ruptured aneurysm of the anterior communicating artery 2 days prior, suddenly developed refractory hypernatremia (serum sodium (Na) 156 - 162 mmol/L). Symptoms included low plasma vasopressin, fluctuating urine osmolality (120 - 710 mOsm/kg) and lack of thirst, all suggesting adipsic diabetes insipidus (ADI). Hypernatremia was corrected by scheduled water intake with desmopressin administration. During 1-year follow-up after the surgery, his serum Na level normalized despite the suspension of desmopressin, but neither thirst nor osmolality-dependent vasopressin release recovered. Meanwhile, his urine osmolality shifted to a constant high level. The present case suggests that renal compensatory adaptation, apparently independent of the circulating vasopressin level, plays a major role in water handling in longitudinal ADI.
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Different Psychosocial Factors Are Associated With Different Intention and Self-Efficacy Toward Eating Breakfast Among Japanese Breakfast Skippers.

A cross-sectional study was conducted among 7593 adults to clarify lifestyle and psychosocial characteristics of breakfast skippers with different intention and self-efficacy toward eating breakfast. Data were obtained from Japan's national survey in 2009. Multinomial logistic regression analyses were performed to examine the characteristics of each breakfast skipper (having intention and self-efficacy toward eating breakfast [IS], having intention but not self-efficacy [INS], and not having intention to eat breakfast [NI]) compared with breakfast eaters. IS men were less likely to demonstrate understanding of healthy eating. INS and NI men were more likely to eat alone. INS and NI women were less likely to demonstrate understanding of healthy eating. NI men and women were less likely to report weight management behavior. Current smoking and being sleep-deprived were commonly associated with breakfast skipping in both sexes. Our results suggested the need for approaches considering type of breakfast skippers (eg, different intention and self-efficacy).

Erratum: A novel heterozygous mutation in the ATP6V0A4 gene encoding the V-ATPase a4 subunit in an adult patient with incomplete distal renal tubular acidosis.

[This corrects the article on p. 424 in vol. 9, PMID: 27274828.].