Potential motivators affecting parental intention in COVID-19 vaccination for children aged 6 months to 4 years: Implications for targeted vaccine interventions in Japan.

Although COVID-19 vaccination was approved for younger children in Japan in October 2022, uptake rates remain critically low. This study aimed to investigate Japanese parents' intentions, hesitators' probability of positive intention change, and factors that motivate COVID-19 vaccination. Parents with a 6-month to 4-year-old child living in Japan participated in this internet-based, cross-sectional survey conducted from December 19, 2022, to January 4, 2023. The modified Poisson regression analysis was used to assess the probabilities of changing intention by each motivator when comparing the degree of hesitancy among hesitators, and the Poisson generalized estimating equations were used to compare the probabilities of changing intentions by potential motivators within hesitant individuals. Among 12,502 participants, 10,008 (80.1%) were hesitators. Parents with lower hesitancy levels were more likely to be motivated to vaccinate their children through potential motivators. Vaccine hesitators were motivated to vaccinate their children, particularly by proven vaccine effectiveness (including "protecting children from getting sick" with a probability ratio [PR] of 3.7 [95% confidence interval (CI) 3.5-3.9] and "less likely to infect adults" with a PR of 2.9 [95% CI 2.8-3.1]), as well as vaccine safety (including "safe vaccination of millions of children" with a PR of 3.1 [95% CI 3.0-3.3]) compared to injunctive norm (including "community leader recommendation"). Therefore, initially addressing parents with low hesitancy levels is an effective strategy that motivates COVID-19 vaccination. Also, providing evidence-based information about COVID-19 vaccine efficacy and safety that is consistent with parents' needs is crucial.

Matrix metalloproteinase-10 deficiency has protective effects against peritoneal inflammation and fibrosis via transcription factor NFκΒ pathway inhibition.

One of the most common causes of discontinued peritoneal dialysis is impaired peritoneal function. However, its molecular mechanisms remain unclear. Previously, by microarray analysis of mouse peritoneum, we showed that MMP (matrix metalloproteinase)-10 expression is significantly increased in mice with peritoneal fibrosis, but its function remains unknown. Chlorhexidine gluconate (CG) was intraperitoneally injected to wild-type and MMP-10 knockout mice to induce fibrosis to elucidate the role of MMP-10 on peritoneal injury. We also examined function of peritoneal macrophages and mesothelial cells obtained from wild-type and MMP-10 knockout mice, MMP-10-overexpressing macrophage-like RAW 264.7 cells and MeT-5A mesothelial cells, investigated MMP-10 expression on peritoneal biopsy specimens, and the association between serum proMMP-10 and peritoneal solute transfer rates determined by peritoneal equilibration test on patients. MMP-10 was expressed in cells positive for WT1, a mesothelial marker, and for MAC-2, a macrophage marker, in the thickened peritoneum of both mice and patients. Serum proMMP-10 levels were well correlated with peritoneal solute transfer rates. Peritoneal fibrosis, inflammation, and high peritoneal solute transfer rates induced by CG were all ameliorated by MMP-10 deletion, with reduction of CD31-positive vessels and VEGF-A-positive cells. Expression of inflammatory mediators and phosphorylation of NFκΒ subunit p65 at S536 were suppressed in both MMP-10 knockout macrophages and mesothelial cells in response to lipopolysaccharide stimulation. Overexpression of MMP-10 in RAW 264.7 and MeT-5A cells upregulated pro-inflammatory cytokines with phosphorylation of NFκΒ subunit p65. Thus, our results suggest that inflammatory responses induced by MMP-10 are mediated through the NFκΒ pathway, and that systemic deletion of MMP-10 ameliorates peritoneal inflammation and fibrosis caused by NFκΒ activation of peritoneal macrophages and mesothelial cells.

ATP6AP2 is robustly expressed in pancreatic ß cells and neuroendocrine tumors, and plays a role in maintaining cellular viability.

ATP6AP2, also known as (pro)renin receptor, has been shown to be expressed in several tissues including pancreatic ß cells. Whereas ATP6AP2 plays an important role in regulating insulin secretion in mouse pancreatic ß cells, the expression profiles and roles of ATP6AP2 in human pancreatic endocrine cells and neuroendocrine tumor cells remain unclear. Here in this study, we investigated the expression profiles of ATP6AP2 in pancreatic endocrine cells, and found that ATP6AP2 is robustly expressed in pancreatic insulinoma cells as well as in normal ß cells. Although ATP6AP2 was also expressed in low-grade neuroendocrine tumors, it was not or faintly detected in intermediate- and high-grade neuroendocrine tumors. Knockdown experiments of the Atp6ap2 gene in rat insulinoma-derived INS-1 cells demonstrated decreased cell viability accompanied by a significant increase in apoptotic cells. Taken together, these findings suggest that ATP6AP2 plays a role in maintaining cellular homeostasis in insulinoma cells, which could lead to possible therapeutic approaches for endocrine tumors.

Dual deletion of guanylyl cyclase-A and p38 mitogen-activated protein kinase in podocytes with aldosterone administration causes glomerular intra-capillary thrombi.

Natriuretic peptides exert not only blood-lowering but also kidney-protective effects through guanylyl cyclase-A (GC-A), a natriuretic peptide receptor. Signaling through GC-A has been shown to protect podocytes from aldosterone-induced glomerular injury, and a p38 mitogen-activated protein kinase (MAPK) inhibitor reduced glomerular injury in aldosterone-infused podocyte-specific GC-A knockout mice. To explore the role of p38 MAPK in podocytes, we constructed podocyte-specific p38 MAPK and GC-A double knockout mice (pod-double knockout mice). Unexpectedly, aldosterone-infused and high salt-fed (B-ALDO)-treated pod-double knockout mice resulted in elevated serum creatinine, massive albuminuria, macrophage infiltration, foot process effacement, nephrin and podocin reduction, and additionally, intra-capillary fibrin thrombi, indicating endothelial injury. Microarray analysis showed increased plasminogen activator inhibitor-1 (PAI-1) in glomeruli of B-ALDO-treated pod-double knockout mice. In B-ALDO-treated pod-double knockout mice, PAI-1 increased in podocytes, and treatment with PAI-1 neutralizing antibody ameliorated intra-capillary thrombus formation. In vitro, deletion of p38 MAPK by the CRISPR/Cas9 system and knockdown of GC-A in human cultured podocytes upregulated PAI-1 and transforming growth factor- ß1 (TGF-ß1). When p38 MAPK knockout podocytes, transfected with a small interfering RNA to suppress GC-A, were co-cultured with glomerular endothelial cells in a transwell system, the expression of TGF-ß1 was increased in glomerular endothelial cells. PAI-1 inhibition ameliorated both podocyte and endothelial injury in the transwell system signifying elevated PAI-1 in podocytes is a factor disrupting normal podocyte-endothelial crosstalk. Thus, our results indicate that genetic dual deletion of p38 MAPK and GC-A in podocytes accelerates both podocyte and endothelial injuries, suggesting these two molecules play indispensable roles in podocyte function.

Sacubitril/valsartan ameliorates renal tubulointerstitial injury through increasing renal plasma flow in a mouse model of type 2 diabetes with aldosterone excess.

BACKGROUND: Aldosterone has been assumed to be one of aggravating factors in diabetic kidney disease (DKD). Natriuretic peptides/guanylyl cyclase-A/cGMP signalling has been shown to ameliorate aldosterone-induced renal injury in mice. Sacubitril/valsartan (SAC/VAL) is used clinically for chronic heart failure and hypertension, in part by augmenting natriuretic peptide bioavailability. The effects of SAC/VAL on renal pathophysiology including in DKD, however, have remained unclarified. METHODS: Eight-week-old male db/db mice fed on a high-salt diet (HSD) were treated with vehicle or aldosterone (0.2 µg/kg/min), and divided into four groups: HSD control, ALDO (aldosterone), ALDO + VAL (valsartan), and ALDO + SAC/VAL group. After 4 weeks, they were analysed for plasma atrial natriuretic peptide (ANP) levels, renal histology, and haemodynamic parameters including glomerular filtration rate (GFR) by FITC-inulin and renal plasma flow (RPF) by para-amino hippuric acid. RESULTS: The ALDO + SAC/VAL group showed significantly increased plasma ANP concentration and creatinine clearance, and decreased tubulointerstitial fibrosis and neutrophil gelatinase-associated lipocalin expression compared to ALDO and ALDO + VAL groups. SAC/VAL treatment increased GFR and RPF, and suppressed expression of Tgfb1, Il1b, Ccl2, and Lcn2 genes compared to the ALDO group. The percentage of tubulointerstitial fibrotic areas negatively correlated with the RPF and GFR. CONCLUSION: In a mouse model of type 2 diabetes with aldosterone excess, SAC/VAL increased RPF and GFR, and ameliorated tubulointerstitial fibrosis. Furthermore, RPF negatively correlated well with tubulointerstitial injury, suggesting that the beneficial effects of SAC/VAL could be through increased renal plasma flow with enhanced natriuretic peptide bioavailability.

Hemophagocytic syndrome in a patient with long-term stable pulmonary sarcoidosis with progressive spleen and bone marrow lesion.

An 83-year-old woman with asymptomatic pulmonary sarcoidosis presented to our hospital with fever and malaise for three months. Abdominal CT showed splenomegaly, and bone marrow examination revealed non-caseating granulomas. Pancytopenia was diagnosed due to bone marrow and splenic lesions of sarcoidosis. Steroid pulses were administered, but the patient died without response to treatment. Pathological autopsy results showed non-caseating granulomas and hemophagocytosis in the spleen and bone marrow. This suggested hemophagocytic syndrome, which was not suspected before death, in addition to sarcoidosis. In patients with splenomegaly and pancytopenia with history of pulmonary sarcoidosis, hemophagocytic syndrome should be considered in differential diagnosis.

Effects of Heat-Killed Lactococcus lactis Strain Plasma on Skin Homeostasis-Related Genes and the Skin Microbiome among Healthy Adults: A Randomized Controlled Double-Blind Study.

Lactococcus lactis subsp. lactis strain plasma (LC-plasma) is a bacterial strain that activates plasmacytoid dendritic cells and induces viral resistance genes via the TLR9/MyD88 pathway. We recently showed that oral administration of LC-plasma prevents skin infection by Staphylococcus aureus, possibly by activating skin immunity. In this study, we conducted a double-blind clinical trial to investigate the effect of oral administration of heat-killed LC-plasma on the skin microbiome, gene expression in the skin, and the skin condition of healthy volunteers. Seventy healthy volunteers were randomly assigned to receive either heat-killed LC-plasma or a placebo for eight weeks. Analysis of the skin microbiome by next-generation sequencing suggested that the alpha-diversity of the skin microbiome did not change during the test period in either group. However, the proportion of species that changed significantly during the test period was 10-fold smaller in the LC-plasma group than in the placebo group, suggesting that LC-plasma may maintain the skin microbiome. Quantitative PCR analysis indicated that tight-junction genes, such as CLDN1 and CLDN12, and the antimicrobial peptide gene BD3 were significantly up-regulated in the LC-plasma group but not in the placebo group. Our results suggest that administration of LC-plasma helps to maintain the skin microbiome and that it affects homeostasis-related genes.

Palatable Noodles as a Functional Staple Food Made Exclusively from Yellow Peas Suppressed Rapid Postprandial Glucose Increase.

Legumes are low-carbohydrate food and are abundant in dietary fiber. In order to provide a functional staple food that does not cause a rapid increase in postprandial blood glucose levels, four kinds of legumes were focused on as ingredients. Noodles made from dehulled yellow pea, unshelled yellow pea, chickpea, and lentil were prepared and evaluated as functional staple foods for controlling blood glucose via an in vitro digestion method. We also measured breaking stress and breaking strain using a creep meter, as well as sensory tests on a 9-point hedonic scale. The noodles made from yellow pea had high values for both breaking stress and breaking strain, and was highly regarded in the sensory tests. Therefore, the noodles made from yellow pea on postprandial glucose and insulin response were measured in a randomized double-blind study (n = 12). The results show that noodles made from yellow pea have a low glycemic index (50.4), and have potential as a functional staple food.

Washing with water alone versus soap in maintaining remission of eczema.

BACKGROUND: There is no consensus on the use of soap in skin care for atopic dermatitis in Japan. Thus, this study aimed to evaluate the efficacy of soap to maintain eczema remission in atopic dermatitis patients during the fall-winter period in Japan. METHODS: This assessor-blinded, pragmatic randomized, non-inferiority study enrolled atopic dermatitis patients whose eczema was controlled by regular steroid ointment application less than or equal to 2 days / week (tacrolimus ointment was permitted). For 8 ± 3 weeks, participants washed their upper and lower limbs on one side with soap (soap side) and on the other side with water alone (water side). The primary outcome was an Eczema Area and Severity Index score at week 8 ± 3. RESULTS: Twenty-nine participants were analyzed. The Eczema Area and Severity Index scores at week 8 ± 3 of the water and soap sides were 0.0 (0.0-0.4) and 0.0 (0.0-0.4), respectively (P = 0.18). The difference between both sides was -0.02 (-0.11-0.08), and the limits of the 95% confidence interval did not reach the prespecified non-inferiority margin. The average Patient-Oriented Eczema Measure score was 1.27 ± 1.7 and 1.32 ± 1.8 for the water and soap sides, respectively (P = 0.92). The total number of additional steroid ointment applications was four (0-20) times and six (0-23) times, respectively (P = 0.98). Participants were categorized according to self-assessments of the usefulness of soap, with 2, 24, and 3 participants in the water-effective, invariant, and soap-effective groups, respectively. CONCLUSIONS: For children with controlled atopic dermatitis, washing with water alone was not inferior to washing with soap for maintaining remission of eczema during the fall-winter period in Japan.

Current status of post-operative infections due to antimicrobial-resistant bacteria after digestive tract surgery in Japan: Japan Postoperative Infectious Complications Survey in 2015 (JPICS'15).

PURPOSE: We herein report the findings of the Japan Postoperative Infectious Complication Survey in 2015 (JPICS'15), which evaluated the rate of post-operative infections and colonization due to antimicrobial-resistant (AMR) bacteria after digestive tract surgery. METHODS: This survey by the Japan Society of Surgical Infection included patients undergoing digestive tract surgery at 28 centers between September 2015 and March 2016. Data included patient background characteristics, type of surgery, contamination status, and type of post-operative infections, including surgical site infections (SSIs), remote infections (RIs), and colonization. RESULTS: During the study period, 7,565 surgeries (of 896 types) were performed; among them, 905 cases demonstrated bacteria after digestive tract surgery. The survey revealed that post-operative infections or colonization by AMR bacteria occurred in 0.9% of the patient cohort, constituting 7.5% of post-operative infections, including 5.6% of SSIs and 1.8% of RIs. Extended-spectrum ß-lactamase-producing Enterobacteriaceae and methicillin-resistant Staphylococcus aureus were the predominant AMR bacteria isolated from patients after digestive tract surgery. Patients infected with AMR bacteria had a poor prognosis. CONCLUSION: Our results reveal that 7.5% of the post-operative infections were due to AMR bacteria, indicating the need for antibacterial coverage against AMR bacteria in patients with critical post-operative infections.

A successful case of percutaneous fistula closure in a patient with high output heart failure and extracardiac arteriovenous fistula.

Intra-cardiac shunt diseases may cause chronic ventricular volume overload, but extra-cardiac fistula could also cause high-output heart failure (HF). A patient presented with high-output HF and significant extra-cardiac shunt flow. Although the size and shape of the patient's left ventricle suggested dilated cardiomyopathy, considerable origins were not identified except for a high-flow fistula between the right subclavian artery and right internal jugular vein. Right heart catheter examination revealed inappropriately high cardiac output. Left-to-right shunt ratio was calculated at 40.3% from an oximetry run, under the assumption that the left anonymous vein which was not contaminated with any shunt flow could be substituted for venous return from the upper body. We could determine the indication of fistula closure according to the estimated high left-to-right shunt ratio, reducing cardiac output by 42.7% which was similar to the pre-estimated left-to-right shunt ratio. Two months later, the patient's serum B-type natriuretic peptide level and left ventricular end-diastolic and end-systolic diameters were decreased. The proposed method to estimate the left-to-right shunt ratio was useful in determining the indication for fistula closure in a patient with HF and a significant shunt fistula. .

The evaluation of frequency of nephrotoxicity caused by liposomal amphotericin B.

BACKGROUND: Liposomal amphotericin B (L-AmB) was developed to reduce nephrotoxicity and maximize the therapeutic utility of amphotericin B in the treatment of invasive fungal infections. However, there is little investigation into the safety of L-AmB in patients with several renal functions. Therefore, we retrospectively evaluated the clinical safety of L-AmB among patients with several renal functions. METHODS: We divided patients treated with L-AmB from April 2014 to September 2016 into 4 groups (estimated glomerular filtration rate (eGFR)≥60, 60 > eGFR≥30, eGFR<30 and hemodialysis). The main endpoint was the incidence of nephrotoxicity and the difference in the serum creatinine values at the end of L-AmB treatment as compared with baseline. RESULTS: The incidence of nephrotoxicity was not significantly different among four groups (eGFR≥60; 27.0%, 60 > eGFR≥30; 30.8%, eGFR<30; 50.0%, hemodialysis; 40.0%, p = 0.56).Only one group of patients with eGFR≥60 admitted the significant increase of serum creatinine value after L-AmB treatment started (p < 0.01). Patients admitted 0.5 mg/dL or more of increase in serum creatinine values until 9 days from the L-AmB therapy started (eGFR≥60; 5.0 days [3.0-8.0 days], 60 > eGFR≥30; 5.0 days [4.0-9.0 days], eGFR<30; 4.5 days [3.0-5.0 days], hemodialysis; 5.5 days [4.0-7.0 days], p = 0.46). CONCLUSION: Take previous clinical study results together, our data suggested that L-AmB is safer agent than amphotericin B for the treatment of fungal infections in patients with eGFR<60 and hemodialysis patients at the start of treatment. Also, especially, we should use L-AmB more carefully until 9 days from the treatment started.

The Prophylactic Effect of Anti-influenza Agents for an Influenza Outbreak in a University Hospital.

Objective From November 24 to December 9, 2013, an outbreak of the influenza (flu) A (H3) virus occurred in a tertiary-care university hospital (1,014 beds). We herein report the prophylactic effect of anti-flu agents for controlling the flu outbreak. Methods We administered pre- or post-exposure prophylaxis with anti-flu agents in flu outbreak. To test the effectiveness of prophylaxis in a flu outbreak, we used the posterior mean of the reproductive value during the pre- and post-intervention period. We also simulated the probability distribution of new flu cases. We performed an analysis to quantify the strength of the intervention effect. Results A total of 97 people were diagnosed with flu before the intervention, and 7 were diagnosed after the intervention. A molecular analysis of the flu virus revealed that this outbreak was due to the flu A (H3) virus. A total of 3,702 people received prophylaxis. There was a significant reduction in the reproductive value from 1.89 [95% confidence interval (CI), 1.59 to 2.24] to 0.65 (95% CI, 0.02 to 1.00) after the intervention (p<0.001). Conclusion Prophylaxis with anti-flu agents, along with prompt identification and isolation of infected individuals, was effective in reducing the impact of a flu outbreak in a hospital.

Enhancement of immunomodulative effect of lactic acid bacteria on plasmacytoid dendritic cells with sucrose palmitate.

Plasmacytoid dendritic cells (pDCs) play a key role in the immune response against viruses. In addition, recent research has suggested that pDCs possess direct and indirect tumoricidal activities. We previously found that a lactic acid bacteria strain, Lactococcus lactis JCM 5805 (LC-Plasma), stimulated pDCs and prevented viral infection in mouse and human studies. Meanwhile, emulsifiers have recently been highlighted as candidate adjuvants for some viral vaccines and cancer immunotherapies. In this study, we discovered some specific emulsifiers, mainly consisting of sucrose fatty acid esters, that drastically enhance the potency of LC-Plasma to activate pDCs in vitro. The emulsifiers promoted the efficient uptake of LC-Plasma by pDCs and the ratio of pDCs that took up LC-Plasma correlated with the activity of pDCs. In addition, an in vivo study showed that oral treatment with LC-Plasma mixed with an emulsifier induced a higher expression of genes related to anti-viral immunity in the lung compared to treatment with LC-Plasma alone. Both LC-Plasma and the emulsifiers used in this study have been confirmed to be safe for human use. Therefore, LC-Plasma mixed with an emulsifier might be a useful tool for certain anti-cancer and anti-viral therapies.

Impact of mucosal barrier injury laboratory-confirmed bloodstream infection (MBI-LCBI) on central line-associated bloodstream infections (CLABSIs) in department of hematology at single university hospital in Japan.

BACKGROUND: Central line-associated bloodstream infections (CLABSIs) are among the most serious complications especially in blood cancer patients. In January 2013, Centers for Disease and Prevention (CDC) introduced a new surveillance definition of mucosal barrier injury-associated laboratory-confirmed bloodstream infection (MBI-LCBI). This study was to determine the impact of MBI-LCBI on CLABSIs and compare the clinical characteristics of MBI versus non-MBI-LCBI cases. PATIENTS AND METHODS: We retrospectively reviewed the records of 250 consecutive patients. They were admitted in department of hematology at Aichi Medical University Hospital. We applied the revised 2013 CLABSI surveillance protocol to all CLABSI cases identified during the 47-months period from May 2012 through June 2016. RESULTS: A total of 44 CLABSIs were identified. The median patient age was 65 years (range, 12 to 89). Among 44 patients, 31 patients were diagnosed as leukemia (70.5%) and 12 patients as lymphoma (27.3%). Six patients underwent bone transplantation for leukemia or myelodysplastic syndrome (13.6%). A total of 20 patients (45.5%) were classified as MBI-LCBI and 24 (54.5%) were classified as non-MBI-LCBI. The primary disease type (P = 0.018), neutropenic within 3 days before CLABSI (MBI-LCBI vs. non-MBI-LCBI: 95.0% vs. 26.3%, P = <0.0001), line(s) removed owing to CLABSI (15.0% vs. 54.2%, P = 0.011) and Gram-negative organisms cultured (70.0% vs. 37.5%, P = 0.004) showed significantly difference between the groups. CONCLUSION: Our data showed that MBI-LCBI cases account for 45.5% of the CLABSI cases identified in blood cancer patients, and constituted a significant burden to this high-risk patient population.

Adverse events of prophylactic anti-influenza agents in medical staffs.

BACKGROUND: We undertook a survey to evaluate the compliance and the tolerability of oseltamivir and zanamivir when they were used as post-exposure prophylaxis among the medical staffs in the 2014-2015 seasons to understand a characteristic of adverse events caused by anti-influenza (flu) agents. MATERIALS AND METHODS: During the study period, 540 medical staffs received oseltamivir (75 mg twice a day for 5 days) or zanamivir (twice a day for 5 days) as post-exposure prophylaxis of influenza, respectively. RESULTS: Four hundred eleven medical staffs of 540 medical staffs (76.1%) provided responses to questionnaire investigations. The adverse events caused by oseltamivir were reported by 86 of 382 medical staffs (22.5%). The most frequent adverse events were gastrointestinal adverse events (13.4%), followed by systemic and local diseases (11.8%), diseases of the nervous system (7.9%) and neuropsychiatric adverse events (0.5%). On the other hand, adverse events caused by zanamivir were reported by one (3.4%) of 29 medical staffs. CONCLUSION: Our survey revealed that 22.5% subjects experienced any adverse events due to oseltamivir. And the regimen showed low compliance than we expected. On the other hands, zanamivir showed high adherence with lower incidence of adverse events.

Evaluation of expression and glycosylation status of UGT1A10 in Supersomes and intestinal epithelial cells with a novel specific UGT1A10 monoclonal antibody.

UDP-Glucuronosyltransferases (UGTs) are major phase II drug-metabolizing enzymes. Each member of the UGT family exhibits a unique but occasionally overlapping substrate specificity and tissue-specific expression pattern. Earlier studies have reported that human UGT1A10 is expressed in the gastrointestinal tract at the mRNA level, but the evaluation at the protein level, especially tissue or cellular localization, has lagged behind because of the lack of a specific antibody. In this study, we prepared a monoclonal antibody to UGT1A10 to elucidate the tissue/cellular distribution and interindividual variability of UGT1A10 protein expression. Western blot analysis revealed that the prepared antibody does not cross-react with any other human UGTs. Using this specific antibody, we observed that UGT1A10 protein is expressed in the small intestine but not in the liver or kidney. Immunohistochemical analysis revealed the expression of UGT1A10 protein in epithelial cells of the crypts and villi of the duodenum. In the small intestine microsomes from six individuals, the UGT1A10 protein levels exhibited 16-fold variability. Dopamine 3- and 4-glucuronidation, which is mainly catalyzed by UGT1A10 and by other UGT isoforms marginally, exhibited 50- to 65-fold variability, and they were not correlated with the UGT1A10 protein levels. Interestingly, the enzymatic activities of recombinant UGT1A10 in insect cells that were normalized to the UGT1A10 protein level were markedly lower than those in pooled human small intestine microsomes. Thus, the UGT1A10 antibody we generated made it possible to investigate the tissue/cellular distribution and interindividual variability of UGT1A10 protein expression for understanding the pharmacological and toxicological role of UGT1A10.

Natriuretic peptide receptor guanylyl cyclase-A pathway counteracts glomerular injury evoked by aldosterone through p38 mitogen-activated protein kinase inhibition.

Guanylyl cyclase-A (GC-A) signaling, a natriuretic peptide receptor, exerts renoprotective effects by stimulating natriuresis and reducing blood pressure. Previously we demonstrated massive albuminuria with hypertension in uninephrectomized, aldosterone-infused, and high salt-fed (ALDO) systemic GC-A KO mice with enhanced phosphorylation of p38 mitogen-activated protein kinase (MAPK) in podocytes. In the present study, we examined the interaction between p38 MAPK and GC-A signaling. The administration of FR167653, p38 MAPK inhibitor, reduced systolic blood pressure (SBP), urinary albumin excretion, segmental sclerosis, podocyte injury, and apoptosis. To further investigate the local action of natriuretic peptide and p38 MAPK in podocytes, we generated podocyte-specific (pod) GC-A conditional KO (cKO) mice. ALDO pod GC-A cKO mice demonstrated increased urinary albumin excretion with marked mesangial expansion, podocyte injury and apoptosis, but without blood pressure elevation. FR167653 also suppressed urinary albumin excretion without reducing SBP. Finally, we revealed that atrial natriuretic peptide increased phosphorylation of MAPK phosphatase-1 (MKP-1) concomitant with inhibited phosphorylation of p38 MAPK in response to MAPK kinase 3 activation, thereby resulting in decreased mRNA expression of the apoptosis-related gene, Bax, and Bax/Bcl2 ratio in cultured podocytes. These results indicate that natriuretic peptide exerts a renoprotective effect via inhibiting phosphorylation of p38 MAPK in podocytes.

Crucial Role of Mesangial Cell-derived Connective Tissue Growth Factor in a Mouse Model of Anti-Glomerular Basement Membrane Glomerulonephritis.

Connective tissue growth factor (CTGF) coordinates the signaling of growth factors and promotes fibrosis. Neonatal death of systemic CTGF knockout (KO) mice has hampered analysis of CTGF in adult renal diseases. We established 3 types of CTGF conditional KO (cKO) mice to investigate a role and source of CTGF in anti-glomerular basement membrane (GBM) glomerulonephritis. Tamoxifen-inducible systemic CTGF (Rosa-CTGF) cKO mice exhibited reduced proteinuria with ameliorated crescent formation and mesangial expansion in anti-GBM nephritis after induction. Although CTGF is expressed by podocytes at basal levels, podocyte-specific CTGF (pod-CTGF) cKO mice showed no improvement in renal injury. In contrast, PDGFRα promoter-driven CTGF (Pdgfra-CTGF) cKO mice, which predominantly lack CTGF expression by mesangial cells, exhibited reduced proteinuria with ameliorated histological changes. Glomerular macrophage accumulation, expression of Adgre1 and Ccl2, and ratio of M1/M2 macrophages were all reduced both in Rosa-CTGF cKO and Pdgfra-CTGF cKO mice, but not in pod-CTGF cKO mice. TGF-ß1-stimulated Ccl2 upregulation in mesangial cells and macrophage adhesion to activated mesangial cells were decreased by reduction of CTGF. These results reveal a novel mechanism of macrophage migration into glomeruli with nephritis mediated by CTGF derived from mesangial cells, implicating the therapeutic potential of CTGF inhibition in glomerulonephritis.

Increase of Total Nephron Albumin Filtration and Reabsorption in Diabetic Nephropathy.

The amount of albumin filtered through the glomeruli and reabsorbed at the proximal tubules in normal and in diabetic kidneys is debated. The megalin/cubilin complex mediates protein reabsorption, but genetic knockout of megalin is perinatally lethal. To overcome current technical problems, we generated a drug-inducible megalin-knockout mouse line, megalin(lox/lox);Ndrg1-CreERT2 (iMegKO), in which megalin expression can be shut off at any time by administration of tamoxifen (Tam). Tam administration in adult iMegKO mice decreased the expression of renal megalin protein by 92% compared with that in wild-type C57BL/6J mice and almost completely abrogated renal reabsorption of intravenously injected retinol-binding protein. Furthermore, urinary albumin excretion increased to 175 µg/d (0.46 mg albumin/mg creatinine) in Tam-treated iMegKO mice, suggesting that this was the amount of total nephron albumin filtration. By comparing Tam-treated, streptozotocin-induced diabetic iMegKO mice with Tam-treated nondiabetic iMegKO mice, we estimated that the development of diabetes led to a 1.9-fold increase in total nephron albumin filtration, a 1.8-fold increase in reabsorption, and a significant reduction in reabsorption efficiency (86% efficiency versus 96% efficiency in nondiabetic mice). Insulin treatment normalized these abnormalities. Akita;iMegKO mice, another model of type 1 diabetes, showed equivalent results. Finally, nondiabetic iMegKO mice had a glomerular sieving coefficient of albumin of 1.7×10-5, which approximately doubled in diabetic iMegKO mice. This study reveals actual values and changes of albumin filtration and reabsorption in early diabetic nephropathy in mice, bringing new insights to our understanding of renal albumin dynamics associated with the hyperfiltration status of diabetic nephropathy.