Transdifferentiation of bovine epithelial towards adipocytes in the presence of myoepithelium.

Orchastric changes in the mammary glands are vital, especially during the lactaction. The secretary epithelial cells together with the supporting myoepithelial and stromal cells function cordially to secrete milk.Increase in the number of luminal epithelial cells and a decrease in adipocytes are visible during lactation, whereas the reverse happens in the involution. However, an early involution occurs if the epithelial transdifferente towards adipocytes in lactation period. We aimed to inhibit the adipocyte transdifferentiation of luminal cells by restraining the PPARγ pathway. Linolenic acid (LA) and thiazolidinediones (TZDs) induced adipogenesis in mammary epithelial cells were conducted in monolayer, mixed culture as well as in transwell plate co-culture with mammary myoepithelial cells.Co-culture with myoepithelial cells showed higher adipogenic gene expression in epithelial cells under LA+TZDs treatment. Increase in the expressions of PPARγ, C/EBPα and vimentin in both mRNA as well as protein levels were observed.Whereas,BADGE treatment blocked LA+TZDs induced adipogenesis, as it could not show a significant rise in adipose related markers. Although comparative results were found in both mixed culture and monolayer conditions, co-culture technic found to work better than the others. In summary, antagonizing PPARγ pathway in the presence of myoepithelial cells can significantly reduce the adipogenisis in epithelial cells, suggestingtherapeutic inhibition of PPARγ can be considered to counter early involution or excessive adipogenesis in mammary epithelium in animals.

Healing of full-thickness articular cartilage defects treated with cultured autologous chondrogenic satellite cells isolated from chondral stem cell niche in rabbits.

BACKGROUND: Healing of articular cartilage has remained in question with the use of conventional treatment modalities such as subchondral drilling and microfracture. As demonstrated in the past, adult stem cells retain promising clonogenicity. Therefore, we conducted this study to elucidate the effects of cultured autologous chondrogenic satellite cells (CACSCs) compared with subchondral drilling (SCD) for the repair of full-thickness articular cartilage defects. MATERIALS AND METHODS: We examined CACSCs isolated from the knee of rabbits using flow cytometry for the expression of stemness and chondrocyte-specific factors. Subsequently, we created a full-thickness cartilage defect model with a diameter of 3 mm and depth of 2 mm on the articular surface of trochlear grooves in the left knee of 24 New Zealand white rabbits. Then we drilled subchondrally through the defect in all animals and stuffed the defects with 10-µg/cm(2) collagen scaffolds. In the treatment group, we instilled CACSCs at 5 × 10(6) cells/mL in the collagen scaffold and collected samples on days 15, 30, and 45. RESULTS: The CACSCs revealed significant expression of CD106, CD44, collagen type 2, and aggrecan. In conjunction with SCD, CACSCs improved healing of the articular cartilage defect, as evidenced by the formation of hyaline-like tissue grossly and histologically. The healed tissue also revealed a significant (P < 0.05) increase in the expression of collagen type 2 and aggrecan (by real-time polymerase chain reaction) during the experiment. CONCLUSIONS: In conjunction with SCD, CACSCs may be considered to improve articular cartilage damage.

Carbohydrase inhibition and anti-cancerous and free radical scavenging properties along with DNA and protein protection ability of methanolic root extracts of Rumex crispus.

The study elucidated carbohydrase inhibition, anti-cancerous, free radical scavenging properties and also investigated the DNA and protein protection abilities of methanolic root extract of Rumex crispus (RERC). For this purpose, pulverized roots of Rumex crispus was extracted in methanol (80% and absolute conc.) for 3 hrs for 60℃ and filtered and evaporated with vacuum rotary evaporator. RERC showed high phenolic content (211 µg/GAE equivalent) and strong 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging (IC(50) = 42.86 (absolute methanol) and 36.91 µg/mL (80% methanolic extract)) and reduced power ability. Furthermore, RERC exhibited significant protective ability in H(2)O(2)/Fe(3+)/ascorbic acid-induced protein or DNA damage and percentage inhibition of the HT-29 cell growth rate following 80% methanolic RERC exposure at 400 µg/mL was observed to be highest (10.2% ± 1.03). Moreover, methanolic RERC inhibited α-glucosidase and amylase effectively and significantly (P < 0.05). Conclusively, RERC could be considered as potent carbohydrase inhibitor, anti-cancerous and anti-oxidant.

Characterization of a novel methionine sulfoxide reductase A from tomato (Solanum lycopersicum ), and its protecting role in Escherichia coli.

Methionine sulfoxide reductase A (MSRA) is a ubiquitous enzyme that has been demonstrated to reduce the S enantiomer of methionine sulfoxide (MetSO) to methionine (Met) and can protect cells against oxidative damage. In this study, we isolated a novel MSRA (SlMSRA2) from Micro-Tom (Solanum lycopersicum L. cv. Micro-Tom) and characterized it by subcloning the coding sequence into a pET expression system. Purified recombinant protein was assayed by HPLC after expression and refolding. This analysis revealed the absolute specificity for methionine-S-sulfoxide and the enzyme was able to convert both free and protein-bound MetSO to Met in the presence of DTT. In addition, the optimal pH, appropriate temperature, and Km and Kcat values for MSRA2 were observed as 8.5, 25oC, 352 ± 25 µM, and 0.066 ± 0.009 S(-1), respectively. Disk inhibition and growth rate assays indicated that SlMSRA2 may play an essential function in protecting E. coli against oxidative damage.