Effects of tranilast on the epithelial-to-mesenchymal transition in peritoneal mesothelial cells

BACKGROUND: We investigated the effects of tranilast on epithelial-to-mesenchymal transition (EMT) in an animal model and on the EMT signaling pathway in human peritoneal mesothelial cells (HPMCs).METHODS: We performed in vitro studies (cytotoxicity, cell morphology, and western blot analyses) on HPMCs from human omenta, along with in vivo studies (peritoneal membrane function and morphometric and immunohistochemical analyses) on Sprague Dawley rats. Thirty-two rats were divided into three groups: control (C) group (peritoneal dialysis [PD] catheter but not infused with dialysate), PD group (4.25% glucose-containing dialysate), and PD + tranilast group (4.25% glucose-containing dialysate along with tranilast).RESULTS: In in vitro experiments, transforming growth factor-beta 1 (TGF-β1) increased α-smooth muscle actin and Snail expression and reduced E-cadherin expression in HPMCs. TGF-β1 also reduced cell contact, induced a fibroblastoid morphology, and increased phosphorylation of Akt, Smad2, and Smad3 in HPMCs. Tranilast significantly inhibited TGF-β1-induced EMT and attenuated these morphological changes in HPMCs. In in vivo studies, after 6 weeks of experimental PD, the peritoneal membrane was significantly thicker in the PD group than in the C group. Tranilast protected against PD-induced glucose mass transfer change and histopathological changes in rats.CONCLUSION: Tranilast prevented EMT both in HPMCs triggered with TGF-β1 and in rats with PD-induced peritoneal fibrosis. Thus, tranilast may be considered a therapeutic intervention that enables long-term PD by regulating TGF-β1 signaling pathways.

Wound healing effects of paste type acellular dermal matrix subcutaneous injection

BACKGROUND: Acellular dermal matrix (ADM) helps wound healing by stimulating angiogenesis, acting as a chemoattractant for endothelial cells, providing growth factors, and permitting a substrate for fibroblasts to attach. The current standard for using paste-type ADM (CG Paste) in wound healing is direct application over the wounds. The major concerns regarding this method are unpredictable separation from the wounds and absorption into negative-pressure wound therapy devices. This study aimed to investigate the effects of subcutaneous injection of paste-type ADM on wound healing in rats. METHODS: Full-thickness skin defects were created on the dorsal skin of rats. Eighteen rats were randomly divided into three groups and treated using different wound coverage methods: group A, with a saline dressing; group B, standard application of CG Paste; and group C, injection of CG Paste. On postoperative days 3, 5, 7, 10, and 14, the wound areas were analyzed morphologically. Histological and immunohistochemical tissue analyses were performed on postoperative days 3 and 7. RESULTS: Groups B and C had significantly less raw surface than group A on postoperative days 10 and 14. Collagen fiber deposition and microvessel density were significantly higher in group C than in groups A and B on postoperative days 3 and 7. CONCLUSIONS: This study showed comparable effectiveness between subcutaneous injection and the conventional dressing method of paste-type ADM. Moreover, the injection of CG Paste led to improved wound healing quality through the accumulation of collagen fibers and an increase in microvessel density.

Erratum: Comparative study of fatty liver induced by methionine and choline-deficiency in C57BL/6N mice originating from three different sources / 한국실험동물학회지

One of the authors' names was misprinted.

Erratum: Comparison of doxorubicin-induced cardiotoxicity in the ICR mice of different sources / 한국실험동물학회지

One of the authors' names was misprinted.

Comparative study of fatty liver induced by methionine and choline-deficiency in C57BL/6N mice originating from three different sources / 한국실험동물학회지

Non-alcoholic fatty liver disease (NAFLD) is believed to be the most prevalent liver disease worldwide and a major cause of chronic liver injury. It is characterized by lipid accumulation in the absence of significant alcohol consumption and frequently progresses to steatohepatitis, liver fibrosis, and hepatocellular carcinoma. Although many studies have been conducted to better understand NAFLD since it was first recognized, there are still many gaps in knowledge of etiology, prognosis, prevention and treatment. Methionine-choline deficient (MCD) diet, a well-established experimental model of NAFLD in rodents, rapidly and efficiently produces the clinical pathologies including macrovesicular steatosis and leads to disease progression. In this study, we measured the response to MCD diet in C57BL/6N mice obtained from three different sources; Korea NIFDS, USA, and Japan. We evaluated changes in body weight, food consumption, and relative weights of tissues such as liver, kidney, gonadal white adipose tissue, inguinal white adipose tissue, and brown adipose tissue. These basic parameters of mice with an MCD diet were not significantly different among the sources of mice tested. After 3 weeks on an MCD diet, histopathological analyses showed that the MCD diet induced clear fat vacuoles involving most area of the acinus in the liver of all mice. It was accompanied by increased serum activities of alanine aminotransferase and aspartate aminotransferase, and decreased levels of serum triglyceride and cholesterol. In conclusion, the response of C57BL6N mice originating from different sources to the MCD diet showed no significant differences as measured by physiological, biochemical, and histopathological parameters.

Comparision of doxorubicin-induced cardiotoxicity in the ICR mice of different sources / 한국실험동물학회지

Doxorubicin is a widely used chemotherapeutic agents and is now part of standard therapeutic regimens for a variety of cancers (eg, hematopoietic malignancies and advanced solid tumors of the breast, ovary, thyroid, and bone). However, a potentially lethal and dose-dependent cardiotoxicity that appears within a short time after treatment limits the usage of doxorubicin in cancer patients. Although the mechanism of doxorubicin-induced cardiotoxicity is not completely understood, it is thought that free radical-induced oxidative stress and excessive production of reactive oxygen species are primary drivers of its toxicity. In this study, we compared the doxorubicin-induced cardiotoxicity of ICR mice obtained from three different sources and evaluated the utility of Korl:ICR stock established by the Korean FDA. Because doxorubicin-induced cardiotoxicity is thought to involve the excessive generation of ROS followed by oxidative stress, we determined the representative tissue index of oxidation, lipid peroxidation, and antioxidant, glutathione (GSH), as well as the parameters of heart injury. Doxorubicin treatment successfully induced cardiotoxicity as evidenced by histological examination and serum parameters (eg, levels of LDH and CK activities) in ICR mice. It was accompanied by increased lipid peroxidation and a decrease in both cysteine and GSH, further supporting previous reports that oxidative stress is a potential mechanism of doxorubicin-induced cardiotoxicity. Of interest, we did not observe a significant difference in doxorubicin-induced cardiotoxicity among mice of different origins. Collectively, our results suggest that Korl:ICR strain may be useful in the research of doxorubicin-induced cardiotoxicity.

The Role of Adipose Tissue-Derived Stem Cells in Ischemic Acute Renal Injury in Rats / 대한신장학회지

PURPOSE: The aim of this study is to explore the role of human adipose tissue-derived adult stem cells (ASCs) in repair of acute ischemic renal injury. METHODS: Acute ischemic tubular necrosis was induced in Spraque-Dawley rats (n=15) by clamping bilateral renal pedicles for 40 minutes. Before releasing the clamps, iron labeled stem cells (1x106/0.3 mL) were directly injected to one side of renal cortex and phosphate-buffered saline (PBS) injections were done as a control to the opposite kidney. The examination on both kidney was done after 1, 2 and 4 weeks (n=5 each). The evaluation of the degree of tubular necrosis and the location of stem cells was done by light and electron microscopically. RESULTS: The rats were all alive without rejection. Morphologically, recovery rate was increased up to about 20% in ASCs injection group in 2 weeks-group (p<0.001). Few Prussian-blue positive cells were demonstrated in 2 weeks-group and more cells were present in 4 weeks-group. The cells which were positively stained in Prussian-blue stain were mainly located in peritubular spaces. Electron microscopically, fine iron granules with electron density were found in the cytoplasm of peritubular endothelial cells. CONCLUSION: As a result, the endothelial cells might be differentiated from injected ASCs. It seems that stem cells help repairing the tubular epithelium in acute renal failure model, not by tubulogenesis but by vasculogenesis. This outcome suggests that the possibility of therapeutic use of ASCs without immunosuppression of recipients in acute ischemic renal injury.

Gene Expression Profiles of Uterine Normal Myometrium and Leiomyoma and Their Estrogen Responsiveness In Vitro

BACKGROUND: Uterine leiomyomas are common benign smooth muscle tumors among the reproductive aged-women. The research has been aimed to identify the differentially expressed genes between normal myometrium and leiomyoma and to investigate the effects of E2 on their expression. METHODS: Gene microarray analysis was performed to identify the differentially expressed genes between normal myomerium and leiomyoma. The data was confirmed at protein level by tissue microarray. RESULTS: Gene microarray analysis revealed 792 upregulated genes in leiomyoma. Four genes (tropomyosin 4 [TPM4], collagen, type IV, alpha 2 [COL4alpha2], insulin-like growth factor binding protein 5 [IGFBP5], tripartite motif-containing 28 [TRIM28]) showed the most dramatic upregulation in all leiomyoma samples. Tissue microarray analyses of 262 sample pairs showed significantly elevated expression of TPM4, IGFBP5, estrogen receptor-alpha, and progesterone receptor (PR) protein in leiomyoma from the patients in their forties, COL4alpha2 in the forties and fifties age-groups, and TRIM28 in the thirties age-group. PR, insulin-like growth factor 1 (IGF-1), IGF-1 receptor (IGF-1R) and IGFBP5 were induced by E2 in in vitro culture of tissue explants from which cells migrated throughout the plate. Among these, PR, IGF-1, IGFBP5 genes showed higher expression in tissue compared to cells-derived from tissue in leiomyoma and IGF-1R in leiomyoma cell. CONCLUSIONS: This observation implies the importance of the whole tissue context including the cells-derived from tissue in the research for the understanding of molecular mechanism of leiomyoma. Here, we report higher expression of TRIM28 in leiomyoma for the first time and identify E2-responsive genes that may have important roles in leiomyoma development.

Antifungal Susceptibility Testing of Dermatophytes / 대한의진균학회지

BACKGROUND: A standardized reference method for dermatophytes in vitro susceptibility testing is lacking. However, with increasing variety of drugs available to treat dermatophytosis, the need for a reference method for dermatophytes testing has become apparent. OBJECTIVE: To evaluate a method of quantifying dermatophytes, the standards for broth microdilution method and evaluation of the availability of disk diffusion method in antifungal susceptibility testing for dermatophytes. METHODS: 14 Candida species (sp.), 30 Trichophyton(T.) mentagrophytes, 9 T. raubitschekii and 11 T. rubrum were tested for fluconazole susceptibility by the broth microdilution method and disc diffusion method. Candida sp. was tested according to National Committee for Clinical Laboratory Standards (NCCLS) M27-A and M44-A. Broth microdilution method for T. mentagrophytes, T. raubitschekii and T. rubrum was operated according to NCCLS M38-A. Disk diffusion method for T. mentagrophytes, T. raubitschekii and T. rubrum was tested refer to NCCLS M44-A. RESULTS: The disk diffusion method showed 50% correlation rate with the broth microdilution method for antifungal susceptibility testing for Candida species. The MIC reading point and incubation time of the broth microdilution method for T. mentagrophytes, T. raubitschekii and T. rubrum are Spec-50 and from 7, 6 and 9 days. Relation between the broth microdilution method and disk diffusion method for dermatophytes is poor. CONCLUSIONS: The good method for quantifying dermatophytes is using vortexing only or liquid nitrogen and homogenizer. Standards of MIC reading point and incubation time of microdilution method for dermatophytes are Spec-50 and from 6 to 9 days. It appears that the disk diffusion method is not recommended method for the antifungal susceptibility testing of dermatophytes.

Histopathologic Findings of Horizontal Sections of Normal Scalp from Korean Females / 대한피부과학회지

BACKGROUND: To date there has not been a study of the histologic data of hair pollicles in scalp hairs of healthy Korean female adults. It is better to evaluate follicular structures in horizontal histologic sectioning than in vertical sectioning. OBJECTIVE: To obtain histologic data of hair follicles in scalp hairs in healthy Korean female adults. METHODS: 20 healthy Korean female volunteers with a normal scalp were analyzed. The age range was 22~29 years (mean 25.7+/-2.3). Transverse sections of 4 mm punch biposy specimens from the vertex were examined at various levels from the papillary dermis to the subcutis. Follicular counts of terminal/vellus hairs and anagen/ telogen hair were evaluated. RESULTS: The total number of total hairs, terminal hairs, vellus hairs, average follicular units, and follicular structures/ mm(2) were 17.5+/-2.6, 15.8+/-2.5, 1.8+/-0.7, 9.0+/-1.6, and 1.4+/-0.2 respectively. Terminal hair:vellus hair ratio was 9:1 and percentage of anagen hairs among total hairs was 95%. Hair density in Korean female was found be significantly lower than that of white females and African American females. CONCLUSION: This data can be used as a guideline for interpreting transverse sections of scalp biopsy specimens in Korean female adults.

Histopathologic Findings of Horizontal Sections of Normal Scalp from a Korean Male / 대한피부과학회지

BACKGROUND: The advantage of transverse sectioning over vertical sectioning of scalp biopsies is morphometric analysis of follicular structures. OBJECTIVE: To obtain histologic data of hair follicles from normal scalp hair in a Korean male. METHOD: A total of 22 Korean male volunteers with a normal scalp were included. The age range was 26+/-4.5 (23-29 year-old). Transverse sections of 4mm punch biopsy specimens from the vertex were examined at various levels from the papillary dermis to the subcutis. Follicular counts of terminal/vellus hairs and anagen/telogen hairs were evaluated. RESULTS: The number of total hairs, terminal hairs, vellus hairs, average follicular units, and follicular structures/mm2 were 21.2+/-4.1, 18.8+/-3.7, 2.4+/-1.4, 9.0+/-1.6, and 1.6+/-0.3, respectively. Terminal hair: vellus hair ratio and anagen: telogen hair ratio were 7.8: 1 and 93.7: 6.3, respectively. Hair density in Korean males was found be significantly lower than that of white males, but similar to that of black males. CONCLUSION: This data can be used as a guideline for interpreting transverse sections of scalp biopsy specimens in Korean males.