Nuclear Hormone Receptor Expression in Mouse Kidney and Renal Cell Lines

Nuclear hormone receptors (NHRs) are transcription factors that regulate carbohydrate and lipid metabolism, immune responses, and inflammation. Although several NHRs, including peroxisome proliferator-activated receptor-γ (PPARγ) and PPARα, demonstrate a renoprotective effect in the context of diabetic nephropathy (DN), the expression and role of other NHRs in the kidney are still unrecognized. To investigate potential roles of NHRs in the biology of the kidney, we used quantitative real-time polymerase chain reaction to profile the expression of all 49 members of the mouse NHR superfamily in mouse kidney tissue (C57BL/6 and db/m), and cell lines of mesangial (MES13), podocyte (MPC), proximal tubular epithelial (mProx24) and collecting duct (mIMCD3) origins in both normal and high-glucose conditions. In C57BL/6 mouse kidney cells, hepatocyte nuclear factor 4α, chicken ovalbumin upstream promoter transcription factor II (COUP-TFII) and COUP-TFIII were highly expressed. During hyperglycemia, the expression of the NHR 4A subgroup including neuron-derived clone 77 (Nur77), nuclear receptor-related factor 1, and neuron-derived orphan receptor 1 significantly increased in diabetic C57BL/6 and db/db mice. In renal cell lines, PPARδ was highly expressed in mesangial and proximal tubular epithelial cells, while COUP-TFs were highly expressed in podocytes, proximal tubular epithelial cells, and collecting duct cells. High-glucose conditions increased the expression of Nur77 in mesangial and collecting duct cells, and liver x receptor α in podocytes. These data demonstrate NHR expression in mouse kidney cells and cultured renal cell lines and suggest potential therapeutic targets in the kidney for the treatment of DN

High Glucose Increases Metallothionein Expression in Renal Proximal Tubular Epithelial Cells

Metallothionein (MT) is an intracellular metal-binding, cysteine-rich protein, and is a potent antioxidant that protects cells and tissues from oxidative stress. Although the major isoforms MT-1 and -2 (MT-1/-2) are highly inducible in many tissues, the distribution and role of MT-1/-2 in diabetic nephropathy are poorly understood. In this study, diabetes was induced in adult male rats by streptozotocin, and renal tissues were stained with antibodies for MT-1/-2. MT-1/-2 expression was also evaluated in mProx24 cells, a mouse renal proximal tubular epithelial cell line, stimulated with high glucose medium and pretreated with the antioxidant vitamin E. MT-1/-2 expression was gradually and dramatically increased, mainly in the proximal tubular epithelial cells and to a lesser extent in the podocytes in diabetic rats, but was hardly observed in control rats. MT-1/-2 expression was also increased by high glucose stimulation in mProx24 cells. Because the induction of MT was suppressed by pretreatment with vitamin E, the expression of MT-1/-2 is induced, at least in part, by high glucose-induced oxidative stress. These observations suggest that MT-1/-2 is induced in renal proximal tubular epithelial cells as an antioxidant to protect the kidney from oxidative stress, and may offer a novel therapeutic target against diabetic nephropathy

Ex Vivo Gene Therapy Treats Bone Complications of Mucopolysaccharidosis Type II Mouse Models through Bone Remodeling Reactivation

Mucopolysaccharidosis type II is a disease caused by organ accumulation of glycosaminoglycans due to iduronate 2-sulfatase deficiency. This study investigated the pathophysiology of the bone complications associated with mucopolysaccharidosis II and the effect of lentivirus-mediated gene therapy of hematopoietic stem cells on bone lesions of mucopolysaccharidosis type II mouse models in comparison with enzyme replacement therapy. Bone volume, density, strength, and trabecular number were significantly higher in the untreated mucopolysaccharidosis type II mice than in wild-type mice. Accumulation of glycosaminoglycans caused reduced bone metabolism. Specifically, persistent high serum iduronate 2-sulfatase levels and release of glycosaminoglycans from osteoblasts and osteoclasts in mucopolysaccharidosis type II mice that had undergone gene therapy reactivated bone lineage remodeling, subsequently reducing bone mineral density, strength, and trabecular number to a similar degree as that observed in wild-type mice. Bone formation, resorption parameters, and mineral density in the diaphysis edge did not appear to have been affected by the irradiation administered as a pre-treatment for gene therapy. Hence, the therapeutic effect of gene therapy on the bone complications of mucopolysaccharidosis type II mice possibly outweighed that of enzyme replacement therapy in many aspects.Wada M., Shimada Y., Iizuka S., et al. Ex Vivo Gene Therapy Treats Bone Complications of Mucopolysaccharidosis Type II Mouse Models through Bone Remodeling Reactivation. Molecular Therapy - Methods and Clinical Development, 19, 261. https://doi.org/10.1016/j.omtm.2020.09.012

Development and validation of a new scoring system for prognostic prediction of community-acquired pneumonia in older adults

The discriminative power of CURB-65 for mortality in community-acquired pneumonia (CAP) is suspected to decrease with age. However, a useful prognostic prediction model for older patients with CAP has not been established. This study aimed to develop and validate a new scoring system for predicting mortality in older patients with CAP. We recruited two prospective cohorts including patients aged ≥ 65 years and hospitalized with CAP. In the derivation (n = 872) and validation cohorts (n = 1, 158), the average age was 82.0 and 80.6 years and the 30-day mortality rate was 7.6% (n = 66) and 7.4% (n = 86), respectively. A new scoring system was developed based on factors associated with 30-day mortality, identified by multivariate analysis in the derivation cohort. This scoring system named CHUBA comprised five variables: confusion, hypoxemia (SpO2 ≤ 90% or PaO2 ≤ 60 mmHg), blood urea nitrogen ≥ 30 mg/dL, bedridden state, and serum albumin level ≤ 3.0 g/dL. With regard to 30-day mortality, the area under the receiver operating characteristic curve for CURB-65 and CHUBA was 0.672 (95% confidence interval, 0.607–0.732) and 0.809 (95% confidence interval, 0.751–0.856; P < 0.001), respectively. The effectiveness of CHUBA was statistically confirmed in the external validation cohort. In conclusion, a simpler novel scoring system, CHUBA, was established for predicting mortality in older patients with CAP

Dynamin 1 is important for microtubule organization and stabilization in glomerular podocytes

Dynamin 1 is a neuronal endocytic protein that participates in vesicle formation by scission of invaginated membranes. Dynamin 1 is also expressed in the kidney; however, its physiological significance to this organ remains unknown. Here, we show that dynamin 1 is crucial for microtubule organization and stabilization in glomerular podocytes. By immunofluorescence and immunoelectron microscopy, dynamin 1 was concentrated at microtubules at primary processes in rat podocytes. By immunofluorescence of differentiated mouse podocytes (MPCs), dynamin 1 was often colocalized with microtubule bundles, which radially arranged toward periphery of expanded podocyte. In dynamin 1-depleted MPCs by RNAi, alpha-tubulin showed a dispersed linear filament-like localization, and microtubule bundles were rarely observed. Furthermore, dynamin 1 depletion resulted in the formation of discontinuous, short acetylated alpha-tubulin fragments, and the decrease of microtubule-rich protrusions. Dynamins 1 and 2 double-knockout podocytes showed dispersed acetylated alpha-tubulin and rare protrusions. In vitro, dynamin 1 polymerized around microtubules and cross-linked them into bundles, and increased their resistance to the disassembly-inducing reagents Ca(2+)and podophyllotoxin. In addition, overexpression and depletion of dynamin 1 in MPCs increased and decreased the nocodazole resistance of microtubules, respectively. These results suggest that dynamin 1 supports the microtubule bundle formation and participates in the stabilization of microtubules

担子菌の発酵能による機能性大豆食品の開発

We evaluated the physiological activity of soybean fermented with mushroom mycelia．Without heating，fibrinolytic activity was observed in Schizophyllum commune（1418mm2），Pleurotus cornucopiae（ 1635mm2），Hericium ramosum（1608mm2）and NAPA（Ganodermataceae produced in Thailand） （1284mm2）． The cell-free crude extract form Schizophyllum commune（563mm2）and Pleurotus cornucopiae（ 369mm2）showed the fibrinolytic activity．Concerning the antithrombin activity，the thrombin clotting time was more than 600 seconds for Coriolus versicolor，Pleurotus cornucopiae，Lenzites betulina，Wolfiporia cocos，Schizophyllum commune， NAPA， Hericium erinaceum，Wynnea gigantea， Ramaria botrytis，Cordyceps militaris，Hericium ramosum and Stropharia rugosoannulata. Even after heating，Wolfiporia cocos and Macrolepiota procera showed the clotting time of more than 600 seconds. Compared with the blank（soybeans before fermentation），the total amino acid level increased to 38-fold for Schizophyllum commune and Pleurotus cornucopiae，to 20-fold for Hericium ramosum and to 15-fold for NAPA. In addition，the possible conversion from the glycoside type of isoflavone to the aglycon type was suggested

Comparison of Glass Capillary Plates and Polyethylene Fiber Bundles as Phantoms to Assess the Quality of Diffusion Tensor Imaging

Purpose: The purpose of this study was to evaluate the suitability of two phantoms, one made of capillary plates and the other polyethylene fibers, for assessing the quality of diffusion tensor imaging (DTI).Methods: The first phantom was a stack of glass capillary plates with many parallel micropores (CP). The second phantom was a bundle of polyethylene fiber Dyneema held together with a thermal shrinkage tube (Dy). High resolution multi-shot echo planar imaging (EPI) DTI acquisitions were performed at b-values of 0 and 1000 s/mm2 and diffusion-times (Tdiff) of 37.7 and 97.7 ms on a preclinical 7T MRI scanner. Thirty diffusion-encoding directions were used. The data were used to calculate the fractional anisotropy (FA), mean diffusivity (MD), and angular dispersion (AD). Further acquisitions were performed at b-values from 0 to 8000 s/mm2 in 14 steps with the diffusion gradient applied parallel (axial) and perpendicular (radial) to the Z direction. On the other hand, the data acquired with a 3T MRI scanner were used to confirm that measurements on a clinical machine are consistent with the 7T MRI results.Results: The dependence of FA, MD and AD on Tdiff was smaller for the Dy than for the CPs. The b-value dependent signal attenuations in the axial direction at Tdiff = 37.7 and 97.7 ms were similar for both phantoms. In the radial direction, Dy demonstrated similar b-value attenuation to that of in vivo tissue for bothTdiffs, but the attenuation for the CPs was affected by the change in Tdiff. Parameter estimates were similar for 3T and 7T MRI.Conclusion: The characteristics of the CP indicate that it can be used as a restricted-diffusion dominant phantom, while the characteristics of the Dy suggest that it can be used as a hindered-diffusion dominant phantom. Dy may be more suitable than CP for DTI quality control