Preparing and Analyzing Expressed Sequence Tags (ESTs) Library for the Mammary Tissue of Local Turkish Kivircik Sheep.

Kivircik sheep is an important local Turkish sheep according to its meat quality and milk productivity. The aim of this study was to analyze gene expression profiles of both prenatal and postnatal stages for the Kivircik sheep. Therefore, two different cDNA libraries, which were taken from the same Kivircik sheep mammary gland tissue at prenatal and postnatal stages, were constructed. Total 3072 colonies which were randomly selected from the two libraries were sequenced for developing a sheep ESTs collection. We used Phred/Phrap computer programs for analysis of the raw EST and readable EST sequences were assembled with the CAP3 software. Putative functions of all unique sequences and statistical analysis were determined by Geneious software. Total 422 ESTs have over 80% similarity to known sequences of other organisms in NCBI classified by Panther database for the Gene Ontology (GO) category. By comparing gene expression profiles, we observed some putative genes that may be relative to reproductive performance or play important roles in milk synthesis and secretion. A total of 2414 ESTs have been deposited to the NCBI GenBank database (GW996847-GW999260). EST data in this study have provided a new source of information to functional genome studies of sheep.

The effects of resveratrol on tissue injury, oxidative damage, and pro-inflammatory cytokines in an experimental model of acute pancreatitis.

Acute pancreatitis (AP) is an acute inflammatory condition that results from the digestion of pancreatic tissue by its own enzymes released from the acinar cells. The objective of this study was to investigate the effects of resveratrol on oxidative damage, pro-inflammatory cytokines, and tissue injury involved with AP induced in a rat model using sodium taurocholate (n = 60). There were three treatment groups with 20 rats per group. Groups I and II received 3% sodium taurocholate solution, while group III underwent the same surgical procedure yet did not receive sodium taurocholate. In addition, group II received 30 mg/kg resveratrol solution. Rats were sacrificed at 2, 6, 12, and 24 h time points following the induction of AP. Blood and pancreatic tissue samples were collected and subjected to biochemical assays, Western blot assays, and histopathologic evaluations. Resveratrol did not reduce trypsin levels and prevent tissue damage. Resveratrol prevented IκB degradation (except for 6 h) and decreased nuclear factor-κB (NF-κB), activator protein-1 (AP-1) (except for 24 h), and levels of TNF-α, IL-6 (except for 24 h), and iNOS in the pancreatic tissue at all time points (P < 0.05). Serum nitric oxide (NO) levels were reduced as well (P < 0.05). Thus, we concluded that resveratrol did not reduce trypsin levels and did not prevent tissue injury despite the reduction in oxidative damage and pro-inflammatory cytokine levels detected in this model of AP.

Effects of curcumin on proinflammatory cytokines and tissue injury in the early and late phases of experimental acute pancreatitis.

BACKGROUND & AIMS: Acute pancreatitis (AP) varies from mild to severe necrotizing changes with high mortality. The objective of the current study was to investigate the effects of curcumin on tissue injury and proinflammatory cytokines in the early and late phases of AP. METHODS: AP was induced by sodium taurocholate in rats (n = 140). First group was left untreated. Group II received 100 mg/kg curcumin daily starting 20 days before AP induction. The rats were allocated into 7 sub-groups (n:5) and were sacrificed at 2, 6, 12, 24, 72, 144 and 288 h following the induction of AP. Blood and pancreatic tissue samples were collected for biochemical and histopathologic evaluations and the assessment of protein and mRNA levels, as well. RESULTS: Curcumin decreased total histopathologic scores in comparison with those of the taurocholate group (P < 0.05). Curcumin increased Caspase-3 activity and decreased trypsin activity, while inhibited nuclear factor-κ (NF-κB) at all time points (P < 0.05) and moreover reduced activator protein-1 (AP-1). Curcumin decreased chemokine (except for 288 h), TNF-α (except for 2 and 24 h), IL-6 (except for 2, 6 and 288 h) and iNOS (except for 144 and 288 h) mRNA levels (P < 0.05). Curcumin serum nitric oxide (NO) (except for 144 and 288 h) levels were reduced, as well. CONCLUSIONS: In conclusion, curcumin reduced tissue injury, trypsin activation and inhibited NF-κB and AP-1. However TNF-α, IL-6 and iNOS and NO were not inhibited at all time points. Therefore no direct correlation was detected in the subgroups between tissue injury, proinflammatory cytokines and oxidative enzymes.

The use of platelet-rich fibrin in combination with biphasic calcium phosphate in the treatment of bone defects: a histologic and histomorphometric study.

BACKGROUND: Platelet-rich fibrin (PRF) is a leukocyte and platelet concentrate containing many growth factors. Its potential for hard tissue augmentation as a sole grafting material or in combination with other grafting materials has been investigated in many studies. OBJECTIVE: The aim of this histologic study was to evaluate the efficacy of PRF mixed with biphasic calcium phosphate (BCP) on bone regeneration in surgically created bone defects. METHODS: Defects 5 mm in diameter were created in both tibias of 6 sheep. The defects were left empty or grafted with BCP, PRF, or BCP+PRF. Animals were killed at 10, 20, and 40 days. The specimens underwent histologic and histomorphometric analysis. RESULTS: None of the groups displayed any signs of necrosis. Inflammation was observed in all groups at 10 days; 2 specimens of PRF+BCP and all empty defects showed inflammatory cell infiltration at 20 days. During the 40-day evaluation period, the PRF+BCP group showed the highest ratios of new bone. The other 3 groups showed statistically similar results. In the BCP and PRF+BCP groups, the residual graft ratios were decreased at consecutive time intervals. The difference between the 2 groups was not statistically significant during follow-up. CONCLUSIONS: The current study revealed a histomorphometric increase in bone formation with the addition of PRF to BCP in surgically created defects in sheep tibia.

Pathologic alterations detected in acute pancreatitis induced by sodium taurocholate in rats and therapeutic effects of curcumin, ciprofloxacin and metronidazole combination.

BACKGROUND AND AIMS: Secondary bacterial infections and free radical injury have been known to play an important role in the pathogenesis and clinical outcome of acute pancreatitis. Despite the therapy models developed in recent years, the mortality rate is still reported to be higher than expected. The objective of this study therefore was to investigate the effectiveness of ciprofloxacin and metronidazole combination and curcumin together in the treatment of acute pancreatitis. METHODS: Acute pancreatitis was induced in rats by sodium taurocholate (n = 60). Starting 6 h after the induction of acute pancreatitis, groups I and II were injected 200 mg/kg ciprofloxacin and 500 mg/kg metronidazole intraperitoneally every 12 h for 6 days. Groups II and III received 100 mg/kg curcumin since day 20 prior to the initiation of acute pancreatitis. On day 6, animals of all groups were killed. Blood and tissue samples were taken for biochemical, pathologic and bacteriologic examination. RESULTS: No statistical difference in the treatment groups versus the non-treatment group has been detected in the pancreatic tissue on the basis of histopathological scoring results. Prevalences of bacterial translocation were significantly lower in the treatment groups (groups I-III) than in the non-treatment group (group IV) (p < 0.001, p < 0.001, p < 0.05, respectively). Serum amylase, lipase, malon dialdehyde and nitric oxide (except for nitric oxide level in group I), levels of groups I, II and III were significantly lower than those of group IV (p < 0.05). CONCLUSIONS: The administration of ciprofloxacin and metronidazole in combination and curcumin in acute pancreatitis failed to provide a preventive effect on the occurrence of tissue injury, whereas free radical injury and prevalence of bacterial translocation were reduced significantly.