Interferon-γ enhances the therapeutic effect of mesenchymal stem cells on experimental renal fibrosis.

Mesenchymal stem cells (MSCs) administered for therapeutic purposes can be activated by interferon-γ (IFN-γ) secreted from natural killer cells in injured tissues and exert anti-inflammatory effects. These processes require a substantial period of time, leading to a delayed onset of MSCs' therapeutic effects. In this study, we investigated whether pretreatment with IFN-γ could potentiate the anti-fibrotic ability of MSCs in rats with ischemia-reperfusion injury (IRI) and unilateral ureter obstruction. Administration of MSCs treated with IFN-γ strongly reduced infiltration of inflammatory cells and ameliorated interstitial fibrosis compared with control MSCs without IFN-γ treatment. In addition, conditioned medium obtained from IFN-γ-treated MSCs decreased fibrotic changes in cultured cells induced by transforming growth factor-ß1 more efficiently than that from control MSCs. Most notably, secretion of prostaglandin E2 from MSCs was significantly increased by treatment with IFN-γ. Increased prostaglandin E2 in conditioned medium obtained from IFN-γ-treated MSCs induced polarization of immunosuppressive CD163 and CD206-positive macrophages. In addition, knockdown of prostaglandin E synthase weakened the anti-fibrotic effects of MSCs treated with IFN-γ in IRI rats, suggesting the involvement of prostaglandin E2 in the beneficial effects of IFN-γ. Administration of MSCs treated with IFN-γ might represent a promising therapy to prevent the progression of renal fibrosis.

High-normal albuminuria is associated with subclinical atherosclerosis in male population with estimated glomerular filtration rate ≥60 mL/min/1.73 m2: A cross-sectional study.

BACKGROUND: Low-grade albuminuria has been considered a predictor of cardiovascular mortality. We investigated the relationship between high-normal albuminuria and subclinical atherosclerosis in non-diabetic men with estimated glomerular filtration rate (eGFR) ≥60 mL/min/1.73 m2. METHODS: In this cross-sectional study, 1,756 men with eGFR ≥60 mL/min/1.73 m2 and urine albumin-to-creatinine ratio (UACR) <30 mg/g, who attended general health checkups between April 2012 and March 2015, underwent blood sampling, urinalysis, and carotid ultrasonography. We excluded the subjects who were diabetic and/or received an anti-hypertensive drug. Carotid intima-media thickness (IMT) and the number of focal atheromatous plaques were used as indicators of subclinical atherosclerosis. Multiple linear regression analysis was performed to identify clinical factors associated with carotid IMT. Poisson regression analysis was used to assess the determinants of the carotid plaque number. RESULTS: Median UACR was 4.8 mg/g (interquartile range, 3.6-6.9 mg/g). Compared with subjects with low-normal UACR (<10.0 mg/g), subjects with high-normal UACR (10.0-29.8 mg/g) had greater IMT and higher carotid plaque number. High-normal UACR was independently associated with thickened IMT in the model adjusted for conventional cardiovascular disease risk factors. Moreover, participants with high-normal UACR were also more likely to be associated with increased plaque count (prevalence ratio: 1.06; 95% confidence interval: 1.01-1.14) after adjustment for conventional cardiovascular disease risk factors. CONCLUSIONS: Our results indicate that high-normal albuminuria is associated with both carotid IMT and plaque formation in the non-diabetic male population with eGFR ≥60 mL/min/1.73 m2.

The role of cyclophilin D in interspecies differences in susceptibility to hepatotoxic drug-induced mitochondrial injury.

Test compound A ((5Z)-6-[(2R,3S)-3-({[(4-Chloro-2-methylphenyl)sulfonyl]amino}methyl) bicyclo[2.2.2]oct-2-yl]hex-5-enoic acid) was withdrawn from premarketing clinical trials due to severe liver injury. Intracellular accumulation of lipids (steatosis) has been observed in human-derived cells and may account for the severe hepatotoxicity. Mitochondrial ß-oxidation and ketogenesis play a fundamental role in energy homeostasis. Mitochondrial dysfunction can therefore cause severe deficiency in fatty acid oxidation and apoptosis which finally triggers the hepatocellular injury. Some of hepatotoxic drugs (e.g., salicylic acid, diclofenac and troglitazone) are known to induce mitochondrial dysfunction. This study therefore examined the effect of compound A on the mitochondrial permeability transition (MPT) and membrane potential in mitochondria isolated from mouse, rat and monkey livers. The incubation of rat and monkey mitochondria energized by succinate in the presence of Ca(2+) (20µM) and compound A (2.5-10µM) resulted in cyclosporin A (CsA)-sensitive MPT pore opening and a decline in mitochondrial membrane potential in a concentration-dependent manner. However, mouse mitochondria showed low susceptibility to compound A-induced dysfunction. Rat mitochondrial expression of cyclophilin D (CyPD) was about twice that of mouse mitochondria, but the expression levels of other MPT pore proteins (adenine nucleotide translocator and voltage-dependent anion channel) were comparable in both species. An assessment of the effect of compound A on CyPD knockdown cells demonstrated that mitochondrial susceptibility to compound A was attenuated in CyPD knockdown cells. These results suggest that an interspecies difference in the susceptibility to mitochondrial dysfunction induced by compound A exists as a result of species-specific discrepancies in CyPD expression.

Treatment of velopharyngeal inadequacy in a patient with submucous cleft palate and myasthenia gravis.

OBJECTIVE: To describe the clinical course and management of a patient with submucous cleft palate who developed myasthenia gravis (MG) as an adult and suffered recurrent hypernasality. Few reports have described MG patients undergoing pharyngeal flap surgery for velopharyngeal incompetence, and these have described only slight speech improvement in such patients. DESIGN: Case report. PATIENT: The patient underwent primary pushback palatoplasty and superiorly based pharyngeal flap surgery for submucous cleft and short palate at age 7. Hypernasality showed major improvement after initial surgery. At age 19, the patient developed MG that triggered the recurrence of velopharyngeal incompetence. INTERVENTION: After MG was treated, revision pushback palatoplasty was performed for velopharyngeal incompetence when the patient was 24 years old. Preoperatively and postoperatively, the patient was evaluated by the same speech-language-hearing therapists, each with at least 5 years of clinical experience in cleft palate speech. RESULTS: After the second pushback palatoplasty, hypernasality and audible nasal air emission during speech decreased to mild. CONCLUSION: Primary pushback palatoplasty and pharyngeal flap surgery were performed for the submucous cleft palate. Revision pushback palatoplasty improved velopharyngeal inadequacy induced by MG. Decreased perceived nasality positively influenced the patient's quality of life. Combined pushback palatoplasty and pharyngeal flap surgery is thus an option in surgical treatment for velopharyngeal inadequacy to close the cleft and the velopharyngeal orifice in cases of cleft palate and MG.

Labeled acetate incorporation into lipids and lipid elimination after oral administration in rat liver and adipose tissue.

To investigate the incorporation of acetate into fatty acids and their turnover, the time courses for the incorporation of labeled acetate into lipids in the liver and epididymal adipose tissue (adipose tissue) after the oral administration to rats were examined for 10 d. The labeled acetate was abundantly incorporated into lipids, mainly into triacylglycerols (TAG) in the liver, reached a maximum at 2 h after the administration and then quickly decreased. In the adipose tissue, the incorporation of the acetate reached a maximum after 8 h and began to decrease slowly after 2 d. The acetate incorporation into the lipids was markedly lower in the liver, plasma and adipose tissue of rats fed the corn oil diet than in those fed the fat-free diet. However, the half-lives of esterified fatty acids were similar in both dietary groups. The half-lives of esterified C16:0 and C18:1 in the decreasing phase were 5.4 and 8.9 h, respectively, in the liver, and 4.3 and 5.6 d, in the adipose tissue. The time courses for incorporation into plasma lipids were parallel to those in the liver. Thus the fatty acids synthesized in the liver appeared to be transported to adipose tissues and to stay there longer. Moreover, it is remarkable that 30% of the acetate radioactivities administered were found after 2 h in the whole liver: 75% of the products from the acetate at the maximum were lipids and 61%, of the lipids, TAG. The major products from acetate in the liver were lipids.

Differences in labelled triolein turnover after oral administration between liver and adipose tissue of rats.

To investigate exogenous triacylglycerol turnover, the time courses for labelled triolein in the liver, plasma and epididymal adipose tissue (adipose tissue) after oral administration to rats fed a fat-free or 10 % corn oil diet for 3 d after fasting overnight were examined for 10 d. After the administration of labelled triolein to rats fed the fat-free diet, the incorporation (dpm/g) into total lipids of the liver and adipose tissue each reached the maximum in 8 h and was seven times higher in the adipose tissue than in the liver. The half-lives of total lipid radioactivities during the decreasing phases were 0.39 and 2.58 d, respectively, in the rapid and slow phases of the decay curve in the liver, and 4.78 d in only one phase of the adipose tissue. Radioactivity after administration of labelled triolein was mostly found in the oleic acid in the tissues. The half-life of oleic acid was 3.92 d in the adipose tissues. These half-lives were similar in both dietary groups. Thus, although dietary corn oil reduced the triolein incorporation to cellular lipids in comparison to the fat-free diet, it did not affect these half-lives. The labelled triacylglycerol-oleic acid stayed abundantly intact for a long time in the adipose tissue and was scarcely changed to other fatty acids, whereas it was slightly incorporated into total lipids and quickly metabolized in the liver. Non-essential fatty acids may be mostly endogenous in the liver but may be exogenous and endogenous in adipose tissue.

Expression of RANTES mRNA in skin lesions of feline eosinophilic plaque.

One of the mechanisms of eosinophil infiltration is its induction by chemoattractants such as regulated upon activation, normal T-expressed and secreted (RANTES) which is a cysteine-cysteine chemokine that mediates chemotaxis and activation of eosinophils in humans and mice. Skin lesions of feline eosinophilic plaque are characterized by a predominant infiltration of eosinophils. The mechanism(s) of eosinophilic infiltration in the skin and/or mucosa of cats is unknown. It is possible that RANTES is involved. To investigate the presence of RANTES in the skin of cats with eosinophilic plaques and nonaffected skin, we cloned and sequenced the full-length feline RANTES cDNA gene, in order to determine whether it is present in the skin of cats with eosinophilic plaques and/or if it is present in normal adjacent skin. We were able to document the the expression of RANTES mRNAs in skin with feline eosinophilic plaque as well as in normal cat skin. The full-length cDNA sequence of the RANTES gene (742 bp) contained a single open reading frame of 276 bp encoding a protein of 92 amino acids. The amino acid sequence of feline RANTES shared 67 and 74% sequence identity with that of bovine and mouse RANTES genes, respectively. RT-PCR analysis on RANTES mRNA in the skin of cats with eosinophilic plaque revealed that its expression was higher in the eosinophilic plaque skin lesions than in the normal skin. The result suggested that RANTES might play a role to induce eosinophil infiltration in feline eosinophilic plaque lesions.

Comparisons of glucose and lipid metabolism in rats fed diacylglycerol and triacylglycerol oils.

The effects of dietary 1,3-diacylglycerol-rich oil (DG oil) on biochemical findings related to glucose and lipid metabolisms were investigated in comparison with triacylglycerol oil (TG oil) in normal rats. Young (7 wk-old) and old (8 mo-old) rats were fed a synthetic diet containing 10% (by weight) DC or TG oil for 1, 4, 8, or 12 wk. The body weights, epididymal and perirenal adipose tissue weights, and feed efficiency were not significantly different in the dietary oil groups during any feeding period. The plasma and liver triacylglycerol concentrations were not different in the dietary groups, except that the plasma triacylglycerol concentrations were rather lower only in the portal vein of rats fed DG oil. The plasma glucose and free fatty acid concentrations were significantly higher in rats fed DG oil as compared to TG oil. In the old rats fed DG oil for 8 wk, the fasted plasma glucose and insulin concentrations were elevated and glucose intolerance was observed. The insulin receptor expression was not different due to dietary oil, but was markedly reduced with aging. Thus, the anti-obesity and lipid-lowering effects of dietary DG oil were not found. Moreover, it appeared that the glucose intolerance might be induced by dietary DG oil, particularly in the old rats.

Dietary diacylglycerol-rich oil stimulation of glucose intolerance in genetically obese rats.

The effects of dietary 1,3-diacylglycerol-rich oil (DG oil) on glucose and lipid metabolism were investigated in comparison with triacylglycerol (TG) oil in female genetically obese Wistar fatty rats. The obese rats and their lean littermates (8 wk old) were fed a synthetic diet containing 10%, (w/w) DG or TG oil for 5 wk. The body weights, abdominal fat weights, and the plasma and liver TG concentrations were not significantly different due to dietary fat type in the obese and lean rats. The plasma glucose concentrations were significantly elevated by dietary DG oil as compared to TG oil in the portal vein and inferior vena cava of obese and lean rats. The plasma free fatty acid concentrations were markedly elevated by dietary DG oil as compared to TG oil in the portal vein and inferior vena cava of both genotype rats, particularly in the obese rats. In the glucose tolerance test, the obese rats fed DG oil showed glucose intolerance, possibly due to the markedly elevated plasma free fatty acids. Thus, the effects of dietary DG oil on lipid-lowering effects and anti-obesity were not observed in either genotype in the present study. Moreover, it is remarkable that glucose intolerance was induced by dietary DG oil in the genetically obese rats. dietary