BACKGROUND: Muscle wasting in sepsis is associated with increased proteolysis. Interleukin-15 (IL-15) has been characterized as an anabolic factor for skeletal muscles. Our study aims to investigate the role of IL-15 in sepsis-induced muscle atrophy and proteolysis. METHODS: Mice were rendered septic either by cecal ligation and puncture or by intraperitoneal injection of lipopolysaccharide (LPS, 10 mg/kg i.p.). Expression of IL-15 mRNA and protein was determined by reverse transcriptase polymerase chain reaction and Western blot analysis in the control and septic limb muscles. C2C12 skeletal muscle cells were stimulated in vitro with either LPS or dexamethasone in the presence and absence of IL-15 and sampled at different time intervals (24, 48, or 72 hours). IL-15 (10microg/kg) was intraperitoneally administered 6 hours before sepsis induction and limb muscles were sampled after 24 hours of sepsis. Cathepsin L activity was determined to measure muscle proteolysis. Atrogin-1 and muscle-specific ring finger protein 1 (MuRF1) expressions in limb muscle protein lysates was analyzed. RESULTS: IL-15 mRNA expression was significantly lower in the limb muscles of septic mice compared to that of controls. Cathepsin L activity in C2C12 cells was significantly lower in presence of IL-15, when compared to that observed with individual treatments of LPS or dexamethasone or tumor necrosis factor alpha. Further, the limb muscles of mice pre-treated with IL-15 prior to sepsis induction showed a lower expression of atrogin-1 and MuRF1 than those not pre-treated. CONCLUSION: IL-15 may play a role in protection against sepsis-induced muscle wasting; thereby, serving as a potential therapeutic target for sepsis-induced skeletal muscle wasting and proteolysis.
Animals ; Atrophy ; Blotting, Western ; Cathepsin L ; Dexamethasone ; Extremities ; Fingers ; Injections, Intraperitoneal ; Interleukin-15 ; Ligation ; Mice ; Muscle Proteins ; Muscle, Skeletal ; Muscles ; Muscular Atrophy ; Proteolysis ; Punctures ; Reverse Transcriptase Polymerase Chain Reaction ; RNA, Messenger ; Sepsis ; Tumor Necrosis Factor-alpha

OBJECTIVE: The efficacy of two-drug combination treatment may be schedule-dependent. We investigated a simulated in-vitro interaction between taxol and doxorubicin in a Cervical cancer cell line HeLa and the role of peroxiredoxin in cytotoxicity. METHODS: Two contradicting schedules of two drugs (taxol followed by doxorubicin or vice versa) were compared each other in terms of cytotoxicity in parental HeLa cell line and the peroxiredoxin (prx)-overexpressing variant. Cytotoxic activity was determined by MTT assay. Cell cycle pertubation was evaluated by flow cytometric analysis. Protein levels were determined by western blot. RESULTS: The sequential treatment of taxol followed by doxorubicin (T-- Appointments and Schedules ; Blotting, Western ; Cell Cycle ; Cell Line ; Cell Line, Tumor* ; Cyclin D1 ; Doxorubicin ; HeLa Cells ; Humans ; Paclitaxel ; Parents ; Peroxiredoxins* ; Uterine Cervical Neoplasms

Age-related rotator cuff tendon degeneration is related to tenofibroblast apoptosis. Anthocyanins reduce oxidative stress-induced apoptotic cell death in tenofibroblasts. The current study investigated the presence of cell protective effects in cyanidin and delphinidin, the most common aglycon forms of anthocyanins. We determined whether these anthocyanidins have antiapoptotic and antinecrotic effects in tenofibroblasts exposed to H₂O₂, and evaluated their biomolecular mechanisms. Both cyanidin and delphinidin inhibited H₂O₂-induced apoptosis in a dose-dependent manner. However, at concentrations of 100 μg/ml or greater, delphinidin showed cytotoxicity against tenofibroblasts and a decreased antinecrotic effect. Cyanidin and delphinidin both showed inhibitory effects on the H₂O₂-induced increase in intracellular ROS formation and the activation of ERK1/2 and JNK. In conclusion, both cyanidin and delphinidin have cytoprotective effects on cultured tenofibroblasts exposed to H₂O₂. These results suggest that cyanidin and delphinidin are both beneficial for the treatment of oxidative stress-mediated tenofibroblast cell death, but their working concentrations are different.
Anthocyanins ; Apoptosis ; Cell Death ; Rotator Cuff ; Tendons

BACKGROUND: Melanogenesis is one of the characteristic parameters of differentiation in melanocytes and melanoma cells. Specific inhibitors of phosphatidylinositol 3-kinase (PI3K), such as wortmannin and LY294002, stimulate melanin production in mouse and in human melanoma cells, suggesting that PI3K and mammalian target of rapamycin (mTOR) might be involved in the regulation of melanogenesis. OBJECTIVE: The involvement of the mTOR pathway in regulating melanogenesis was examined using human MNT-1 melanoma cells, and the effects of the potent inhibitor of mTOR, rapamycin, in the presence or absence of alpha-melanocyte-stimulating hormone (alpha-MSH) were evaluated. METHODS: In cells treated with rapamycin, cell viability, melanin content, and tyrosinase (TYR) activity were measured and compared with untreated controls. Protein levels of TYR, tyrosinase-related protein (TYRP)-1, TYRP-2, and microphthalmia-associated transcription factor (MITF) were also analyzed by Western blot. RESULTS: In rapamycin-treated cells, the melanin content increased concomitantly with an elevation in TYR activity, which plays a major role in melanogenesis. There was also an up-regulation of TYR, TYRP-1, and MITF proteins. Combined treatment with rapamycin or wortmannin and alpha-MSH increased melanogenesis more strongly than alpha-MSH alone. CONCLUSION: Rapamycin-induced melanin formation may be mediated through the up-regulation of TYR protein and activity. Furthermore, rapamycin and wortmannin, inhibitors of mTOR and PI3K, respectively, have co-stimulatory effects with alpha-MSH in enhancing melanogenesis in melanocyte cells.
alpha-MSH ; Androstadienes ; Animals ; Cell Survival ; Chromones ; Humans ; Melanins ; Melanocytes ; Melanoma ; Mice ; Microphthalmia-Associated Transcription Factor ; Monophenol Monooxygenase ; Morpholines ; Phosphatidylinositol 3-Kinase ; Sirolimus ; Up-Regulation

Purpose: We investigated the expression levels of 84 genes in dexamethasone-exposed human lens epithelial cells using polymerase chain reaction (PCR) array analysis. Methods: The viability and motility of lens epithelial cells were examined after treatment with dexamethasone at 0.01, 0.1, and 1 mg/mL; Western blot was used to evaluate the expression levels of fibronectin, α-smooth muscle actin (α-SMA), and E-cadherin. After 24, 48, and 72 hours of dexamethasone treatment at 0.1 mg/mL, the expression levels of 84 growth factors were analyzed using PCR array. Results: Cell viability did not change significantly at dexamethasone levels of 0.01 or 0.1 mg/mL, but decreased markedly at 1 mg/mL; motility increased in a concentration-dependent manner at 0.01 and 0.1 mg/mL. Western blot showed that fibronectin levels increased significantly at all dexamethasone concentrations tested; the α-SMA level increased only at 0.01 mg/mL, and E-cadherin levels decreased significantly at all tested concentrations. PCR showed that the levels of FGF1, FGF2, IL-11, regulators of apoptosis (GDNF, IL-1β, and NRG2), and regulators of cell differentiation (BMP5, FGF1, FGF2, and FGF5) decreased more than twofold, whereas the levels of FGF9 and FGF19 increased more than twofold. Conclusions PCR performed after exposure of lens epithelial cells to dexamethasone may identify the genes involved in the development of steroid-induced cataracts.

Rheumatoid arthritis (RA) is a chronic inflammatiory disease that mainly destroys cartilages or bones at the joints. This inflammatory disorder is initiated by self-attack using own immune system, but the detail of pathological mechanism is unclear. Features of autoantigens leading to autoimmune disease are also under veil although several candidates including type II collagen have been suggested to play a role in pathogenesis. In this report, we tried to identify proteins responding to antibodies purified from RA patients and screen proteins up-regulated or down-regulated in RA using proteomic approach. Fibronectin, semaphorin 7A precursor, growth factor binding protein 7 (GRB7), and immunoglobulin mu chain were specifically associated with antibodies isolated from RA synovial fluids. In addition, some metabolic proteins such as adipocyte fatty acid binding protein, galectin-1 and apolipoprotein A1 precursor were overexpressed in RA synovium. Also, expression of peroxiredoxin 2 was up-regulated in RA. On the contrary, expression of vimentin was severely suppressed in RA synoviocytes. Such findings might give some insights into understanding of pathological mechanism in RA.
Synovial Fluid/metabolism ; Sepharose/chemistry ; Proteomics/methods ; Middle Aged ; Male ; *Inflammation ; Humans ; *Gene Expression Regulation ; Female ; Collagen Type II/biosynthesis ; Autoantigens/metabolism ; Arthritis, Rheumatoid/*metabolism ; Aged ; Adult

Semisulcospira libertina, a species of freshwater snail, is widespread in East Asia. It is important as a food source. Additionally, it is a vector of clonorchiasis, paragonimiasis, metagonimiasis, and other parasites. Although S. libertina has ecological, commercial, and clinical importance, its whole-genome has not been reported yet. Here, we revealed the genome of S. libertina through de novo assembly. We assembled the whole-genome of S. libertina and determined its transcriptome for the first time using Illumina NovaSeq 6000 platform. According to the k-mer analysis, the genome size of S. libertina was estimated to be 3.04 Gb. Using RepeatMasker, a total of 53.68% of repeats were identified in the genome assembly. Genome data of S. libertina reported in this study will be useful for identification and conservation of S. libertina in East Asia.

young adults are sometimes extracted for orthodontic treatment. In the present study, we evaluated the osteogenic activity and mineralization of cultured human dental papilla-derived cells. Dental papillas were harvested from mandible during surgical extraction of lower impacted third molar from 3 patients aged 13-15 years. After passage 3, the dental papilladerived cells were trypsinized and subsequently suspended in the osteogenic induction DMEM medium supplemented with 10% fetal bovine serum, 50 g/ml L-ascorbic acid 2-phosphate, 10 nM dexamethasone and 10 mM -glycerophosphate at a density of 1 x 10(6) cells/dish in a 100-mm culture dish. The dental papilla-derived cells were then cultured for 6 weeks and the medium was changes every 3 days during the incubation period. Dental papilla-derived cells showed positive alkaline phosphatase (ALP) staining during 42 days of culture period. The formation of ALP stain showed its maximal manifestation at day 7 of culture period, then decreased in intensity during the culture period. ALP mRNA level was largely elevated at 1 weeks and gradually decreased with culture time. Osteocalcin mRNA expression appeared at day 14 in culture, after that its expression continuously increased in a time-dependent manner up to day 28. The expression remained constant thereafter. Runx2 expression appeared at day 7 with no detection thereafter. Von Kossa-positive mineralization nodules were first present at day 14 in culture followed by an increased number of positive nodules during the entire duration of the culture period. Osteocalcin secretion was detectable in the culture medium from 1 week. The secretion of osteocalcin from dental papilla-derived cells into the medium greatly increased after 3 weeks although it showed a shallow increase by then. In conclusion, our study showed that cultured human dental papilla-derived cells differentiated into active osteoblastic cells that were involved in synthesis of bone matrix and the subsequent mineralization of the matrix.]]>
Alkaline Phosphatase ; Ascorbic Acid ; Bone Matrix ; Dental Papilla ; Dental Sac ; Dentin ; Dexamethasone ; Enamel Organ ; Humans ; Mandible ; Molar, Third ; Osteoblasts ; Osteocalcin ; RNA, Messenger ; Stem Cells ; Tooth ; Tooth Germ ; Trypsin ; Young Adult

Alkaline Phosphatase ; Calcium ; Cell Culture Techniques ; Cell Proliferation ; Dexamethasone ; Durapatite ; Glucocorticoids ; Humans ; Osteoblasts ; Osteogenesis ; Osteonecrosis ; Osteoporosis ; Periosteum ; Phenotype

Bone and Bones ; Calcium ; Durapatite ; Humans ; Osteoblasts ; Osteocalcin ; Polyethylenes ; Polymers ; Polypropylenes ; RNA, Messenger ; Tissue Engineering ; Transplants